Compounds and methods for modulating gene expression

The present disclosure describes short antisense compounds, including such compounds comprising chemically-modified highaffinity monomers 8-16 monomers in length. Certain such short antisense compound are useful for the reduction of target nucleic acids and/or proteins in cells, tissues, and animals with increased potency and improved therapeutic index. Thus, provided herein are short antisense compounds comprising high-affinity nucleotide modifications useful for reducing a target RNA in vivo. Such short antisense compounds are effective at lower doses than previously described antisense compounds, allowing for a reduction in toxicity and cost of treatment. In addition, the described short antisense compounds have greater potential for oral dosing.

rue Victoria · Place du Portage 1 · Gatineau, (Québec) K1A0C9 · www.opic.ic.gc.ca

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CA 2651309 C 2019/06/11 (11)(21) 2 651 309 (13) C (72) lnventeurs(sulte)/lnventors(continued): SIWKOWSKI, ANDREW M., US; SWAYZE, ERIC E., US;

WANCEWICZ, EDWARD, US (73) Propriétalres(suite)/Owners(continued):IONIS PHARMACEUTICALS, INC., US (74) Agent: NORTON ROSE FULBRIGHT CANADA LLP/S.E.N.C.R.L, S.R.L.

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JUMBO APPLICATIONS / PATENTS

THIS SECTION OF THE APPLICATION / PATENT CONTAINS MORE THAN ONE VOLUME.

THIS IS VOLUME 1 OF 2

NOTE: For additional volumes please contact the Canadian Patent Office.

COMPOUNDS AND METHODS FOR MODULATING GENE EXPRESSION

SEQUENCE LISTING

The present application is being filed along with a Sequence Listing in electronic format. The Sequence Listing is provided as a file entitled CORE0061WO7SEQ.txt, created on May 7, 2007 which is 700 Kb in size. The information in the electronic format of the sequence listing forms part of the 15 present description.

BACKGROUND

Targeting disease-causing gene sequences was first suggested nearly 40 years ago (Belikova et al., Tet. Lett., 1967, 37, 3557-3562), and antisense activity was demonstrated in cell culture a decade 20 later (Zamecnik et al., Proc. Natl. Acad. Sci. U.S A., 1978,75,280-284). One advantage of antisense technology in the treatment of a disease or condition that stems from a disease-causing gene is that it is a direct genetic approach that has the ability to modulate expression of specific disease-causing genes.

Generally, the principle behind antisense technology is that an antisense compound hybridizes to a target nucleic acid and effects modulation of gene expression activity or function, such as transcription, 25 translation or splicing. The modulation of gene expression can be achieved by, for example, target degradation or occupancy-bascd inhibition. An example of modulation of RNA target function by degradation is RNase Il-based degradation of the target RNA upon hybridization with a DNA-likc antisense compound. Another example of modulation of gene expression by target degradation is RNA interference (RNAi). RNAi is a form of antisensc-mediated gene silencing involving the introduction of 30 dsRNA leading to the sequence-specific reduction of targeted endogenous mRNA levels. Sequencespecificity makes antisense compounds extremely attractive as tools for target validation and gene functionalization, as well as research tools for identifying and characterizing nucleases and as therapeutics to selectively modulate the expression of genes involved in the pathogenesis of any one of a variety of diseases.

Antisense technology is an effective means for reducing the expression of one or more specific gene products and can therefore prove to be uniquely useful in a number of therapeutic, diagnostic, and research applications. Chemically modified nucleosides are routinely used for incorporation into

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-2antisense compounds to enhance one or more properties, such as nuclease resistance, pharmacokinetics or affinity for a target RNA.

Despite the expansion of knowledge since the discovery of antisense technology, there remains an unmet need for antisense compounds with greater efficacy, reduced toxicity and lower cost. Until the 5 present disclosure, high-affinity modifications have not been employed in the design of short antisense compounds for reducing target RNA in vivo. This is because of concerns regarding the degree of target specificity that a sequence 15 nucleotides or shorter would have when employed to reduce target in a living system. Previous studies have described that greater specificity, and therefore greater potential for potency, is achieved by antisense compounds between 16 and 20 nucleobases in length.

The present disclosure describes incorporation of chemically-modified high-affinity nucleotides into antisense compounds allows for short antisense compounds about 8-16 nucleobases in length useful in the reduction of target RNAs in animals with increased potency and improved therapeutic index. Thus, provided herein are short antisense compounds comprising high-affinity nucleotide modifications useful for reducing a target RNA in vivo. Such short antisense compounds are effective at lower doses than previously described antisense compounds, allowing for a reduction in toxicity and cost of treatment.

SUMMARY

Disclosed herein are short antisense compounds and uses of said compounds to reduce target RNA expression in cells or tissues. In certain embodiments, provided herein is a use of a short antisense 20 compound targeted to a nucleic acid of a target for reducing expression of such target in an animal. In certain embodiments, short antisense compounds are oligonucleotide compounds. Disclosed are short antisense oligonucleotides of about 8 to 16, preferably 9 to 15, more preferably 9 to 14, more preferably 10 to 14 nucleotides in length and comprises a gap region flanked on each side by a wing, wherein each wing independently consists of 1 to 3 nucleotides.

Preferred motifs include but are not limited to wing - deoxy gap - wing motifs selected from 310-3, 2-10-3, 2-10-2, 1-10-1, 2-8-2, 1-8-1, 3-6-3 or 1-6-1. In a preferred embodimenL the short antisense oligonucleotide comprise at least one high-affinity modification. In a further embodiment, the high-affinity modification includes chemically-modified high-affinity nucleotides. In a preferred embodiment, each wing independently consists of 1 to 3 high-affinity modified nucleotides. In one

30. embodiment the high affinity modified nucleotides are sugar-modified nucleotides.

In one embodiment, there is provided a short antisense compound 10 to 14 monomers in length, comprising a 2’-deoxyribonucleotide gap region flanked on each side by a wing, wherein each wing independently consists of 1 to 3 high-affinity modified monomers which are sugar-modified nucleotides

-2athat comprise a bridge between the 4’ and the 2’ position of the sugar, for use in treating a metabolic disorder in an animal, wherein the sequence of the antisense compound is complementary to an RNA or target gene associated with a metabolic disorder.

In another embodiment, there is provided use of a short antisense compound 10 to 14 monomers in length, comprising a 2’-deoxyribonucleotide gap region flanked on each side by a wing, wherein each wing independently consists of 1 to 3 high-affinity modified monomers which are sugar-modified nucleotides that comprise a bridge between the 4’ and the 2’ position of the sugar, for the preparation of a medicament for treating a metabolic disorder in an animal, wherein the sequence of the antisense compound is complementary to an RNA or target gene associated with a metabolic disorder.

In another embodiment, there is provided a short antisense compound 10 to 14 monomers in length, comprising a 2’-deoxyribonucleotide gap region flanked on each side by a wing, wherein each wing independently consists of 1 to 3 high-affinity modified monomers which are sugar-modified nucleotides that comprise a bridge between the 4’ and the 2’ position of the sugar, for use in treating a cardiovascular disorder in an animal, wherein the sequence of the antisense compound is complementary to an RNA or target gene associated with a cardiovascular disorder.

In another embodiment, there is provided use of a short antisense compound 10 to 14 monomers in length, comprising a 2’-deoxyribonucleotide gap region flanked on each side by a wing, wherein each wing independently consists of 1 to 3 high-affinity modified monomers which arc sugar-modified nucleotides that comprise a bridge between the 4’ and the 2’ position of the sugar, for the preparation of a medicament for treating a cardiovascular disorder in an animal, wherein the sequence of the antisense compound is complementary to an RNA or target gene associated with a cardiovascular disorder.

In another embodiment, there is provided a short antisense compound 10 to 14 monomers in length, comprising a 2'-dcoxyribonuclcotidc gap region flanked on each side by a wing, wherein each wing independently comprises 1 to 3 high-affinity modified nucleotides which are sugar-modified nucleotides that comprise a bridge between the 4’ and the 2’ position of the sugar, and wherein said short antisense compound further comprises a covalently attached conjugate group.

In another embodiment, there is provided a pharmaceutical composition prepared for oral administration comprising an effective amount of a short antisense compound 10 to 14 monomers in length, wherein the short antisense compound comprises a 2'-dcoxyribonucleotidc gap region flanked on each side by a wing, wherein each wing independently comprises 1 to 3 high-affinity modified nucleotides which arc sugar-modified nucleotides that comprise a bridge between the 4’ and the 2’ position of the sugar, and a pharmaceutically acceptable carrier, diluent or excipient.

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-2bIn another embodiment, there is provided a short antisense compound 10 to 14 monomers in length, wherein the short antisense compound comprises a 2'-dcoxyribonucleotide gap region flanked on each side by a wing, wherein each wing independently comprises 1 to 3 high-affinity modified nucleotides which are sugar-modiiïed nucleotides that comprise a bridge between the 4’ and the 2’ position of the sugar, for use in treating a metabolic condition or disorder in a mammalian subject, wherein said short antisense compound is adapted for oral administration and wherein the sequence of the antisense compound is complementary to an RNA or target gene associated with a metabolic disorder.

In another embodiment, there is provided use of a short antisense compound 10 to 14 monomers in length, wherein the short antisense compound comprises a 2'-deoxyribonucleotidc gap region flanked on each side by a wing, wherein each wing independently comprises 1 to 3 high-affinity modified nucleotides which arc sugar-modified nucleotides that comprise a bridge between the 4’ and the 2’ position of the sugar, for the preparation of a medicament for treating a metabolic condition or disorder in a mammalian subject, wherein medicament is adapted for oral administration and wherein the sequence of the antisense compound is complementary to an RNA or target gene associated with a metabolic disorder.

In certain embodiments short antisense compounds exhibit greater uptake in the gut as compared to antisense compounds of greater length. Thus, also provided herein are methods of reducing a target in an animal, comprising orally administering the short antisense compounds of the present invention.

In certain embodiments, short antisense compounds are targeted to a nucleic acid encoding a protein selected from ApoB, SGLT2, PCSK9, SOD1, CRP, GCCR, GCGR, DGAT2, PTP1B and PTEN.

Further provided arc methods of treating a metabolic disorder in an animal, comprising administering to an animal in need of such therapy a short antisense compound targeted to a nucleic acid involved in regulating glucose metabolism or clearance, lipid metabolism, cholesterol metabolism, or

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-3insulin signaling.

Also provided are methods of increasing insulin sensitivity, decreasing blood glucose or decreasing HbA|_C in an animal, comprising administering to said animal a short antisense compound targeted to a nucleic acid encoding a target involved in regulating glucose metabolism or clearance, lipid 5 metabolism, cholesterol metabolism, or insulin signaling.

Further provided are methods of decreasing total serum cholesterol, serum LDL, serum VLDL, serum HDL, serum triglycerides, serum apolipoprotein(a) or free fatty acids in an animal, comprising administering to said animal a short antisense compound targeted to a nucleic acid encoding a target that is involved in regulating glucose metabolism or clearance, lipid metabolism, cholesterol metabolism, or 10 insulin signaling, wherein said short antisense compound is 8 to 16 nucleotides in length and comprises a gap region flanked on each side by a wing, wherein each wing independently consists of 1 to 3 highaffinity modified nucleotides.

Certain targets involved in regulating glucose metabolism or clearance, lipid metabolism, cholesterol metabolism, or insulin signaling include, but are not limited to, GCGR and ApoB-100. Thus, 15 provided are short antisense compounds targeting nucleic acids encoding GCGR and ApoB-100 and methods of reducing expression of said targets and/or target nucleic acids in animal. In addition, provided is the use of short antisense compounds targeting nucleic acids encoding GCGR, and ApoB-100 for die treatment of a metabolic or cardiovascular disease or condition.

In certain embodiments, short antisense compounds further comprise a conjugate group.

Conjugate groups include, but are not limited to, Cu and cholesterol.

In certain embodiments short antisense compounds comprise at least one modified nucleobase, intemucleoside linkage or sugar moiety. In certain embodiments, such modified intemucleoside linkage is a phosphorothioate intemucleoside linkage. In certain embodiments, each intemucleoside linkage is a phosphorothioate intemucleoside linkage.

In certain embodiments, short antisense compounds comprise at least one high affinity modification. In certain such embodiments, the high-affinity modification is a chemically-modified highaffinity nucleotide. In certain embodiments, chemically-modified high affinity nucleotides are sugarmodified nucleotides. In certain embodiments, at least one of the sugar-modified nucleotides comprises a bridge between the 4' and the 2’ position of the sugar. Each of the sugar-modified nucleotides is, 30 independently, in the β-D or α-L sugar conformation. In certain embodiments, each of said high-affinity modified nucleotides confers a T_m of at least 1 to 4 degrees per nucleotide. In certain embodiments, each of said sugar-modified nucleotides comprises a 2'-substituent group that is other than H or OH. Such sugar-modified nucleotides include those having a 4’ to 2’ bridged bicyclic sugar moiety. In certain embodiments, each of the 2'-substituent groups is, independently, alkoxy, substituted alkoxy, or 35 halogen. In certain embodiments, each of the 2'-substituent groups is OCH2CH2OCH3 (2'-MOE).

In certain embodiments, short antisense compounds have one or more sugar-modified nucleotides comprising a bridge between the 4’ and 2’ position of the sugar, wherein each of said bridges

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-4independently comprises from 2 to 4 linked groups independently selected from -[C(R|)(R₂)]_n-, -C(R,)=C(R₂)-, -C(R,)=N-, -C(=NR,)-, -C(=O)-, -C(=S)-> -O-, -Si(R,)₂-, -S(=O)„- and -N(R,)-;

wherein x isO, l,or2;

n is 1,2,3, or 4;

each R| and R₂ is, independently, H, a protecting group, hydroxyl, C₍-Ci₂ alkyl, substituted C|-Cj₂ alkyl, C₂-C|₂ alkenyl, substituted C₂-C|₂ alkenyl, C₂ ne-Cn alkynyl, substituted C₂-C|₂ alkynyl, C₅-C₂₀ aryl, substituted C5-C20 aryl, heterocycle radical, substituted heterocycle radical, heteroaryl, substituted heteroaryl, C5-C7 alicyclic radical, substituted C₅-C₇ alicyclic radical, halogen, OJ|, NJ|J₂, SJ|, N3, COOJ_b acyl (C(=O)-H), substituted acyl, CN, sulfonyl (S(=O)2-J|), or sulfoxyl (S(=O)-J|); and each J| and J₂ is, independently, H, C_rC|₂ alkyl, substituted CpC|₂ alkyl, CrC|₂alkenyl, substituted C₂-C|₂ alkenyl, alkynyl, substituted Q-Cn alkynyl, Cj-Ca) aryl, substituted Cs-C₂q aryl, acyl (C(=O)-H), substituted acyl, a heterocycle radical, a substituted heterocycle radical, Ci-Ct₂ aminoalkyl, substituted C_rC|₂ aminoalkyl or a protecting group.

In one aspect, each of said bridges is, independently, -[C(RiXR₂)]_n-, -[C(Ri)(R₂)J_w-O-, -C(R_fR₂)N(R|)-O- or -C(R|R₂)-O-N(Ri)-. In another aspect, each of said bridges is, independently, 4'-(CH₂)3-2', 4'-(CH₂)₂-2’, 4'-CH₂-O-2', 4'-(CH₂)2-O-2', 4'-CH₂-O-N(Ri)-2’ and 4'-CH_rN(R|)-O-2’- wherein each R, is, independently, H, a protecting group or C|-C|₂ alkyl.

In certain embodiments, provided herein are short antisense compounds useful in the reduction of targets and/or target RNAs associated with disease states in animals. In certain embodiments, provided are methods of using the short antisense compounds for reducing expression of a target RNA in an animal. In certain embodiments, provided herein is the use of a short antisense compound in the preparation of a medicament for the treatment of a metabolic disorder in an animal. In certain embodiments, provided herein is the use of a short antisense compound in the preparation of a medicament for increasing insulin sensitivity, decreasing blood glucose or decreasing HbA|_C in an animal. Also provided is the use of a short antisense compound in the preparation of a medicament for decreasing total serum cholesterol, serum LDL, serum VLDL, serum HDL, serum triglycerides, serum apolipoprotein(a) or free fatty acids in an animal.

In certain embodiments, short antisense compounds provided herein exhibit equal or increased potency with regard to target RNA knockdown as compared to longer parent antisense oligonucleotide at least 20 nucleotides in length. In certain embodiments, short antisense compounds exhibit a faster onset of action (target RNA reduction) as compared to the parent antisense oligonucleotide. In certain embodiments, increased potency is in the kidney. In certain embodiments, target RNA is predominately expressed in the kidney. In certain embodiments, increased potency is in the liver. In certain embodiments, target RNA is predominately expressed in the liver.

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-5DETAILED DESCRIPTION

It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the invention, as claimed. Herein, the use of the singular includes the plural unless specifically stated otherwise. As used herein, the use of or means and/or unless stated otherwise. Furthermore, the use of the term including as well as other forms, such as includes and included, is not limiting. Also, terms such as element or component encompass both elements and components comprising one unit and elements and components that comprise more than one subunit, unless specifically stated otherwise.

The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described. Information relevant to the present disclosure may be found in US patent application serial nos 10/712,795 and 10/200,710.

A. Definitions

Unless specific definitions are provided, the nomenclature utilized in connection with, and the procedures and techniques of, analytical chemistry, synthetic organic chemistry, and medicinal and pharmaceutical chemistry described herein are those well known and commonly used in the art. Standard techniques may be used for chemical synthesis, chemical analysis, pharmaceutical preparation, formulation and delivery, and treatment of subjects. Certain such techniques and procedures may be found for example in Carbohydrate Modifications in Antisense Research Edited by Sangvi and Cook, American Chemical Society , Washington D.C., 1994; and Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pa., 18th edition, 1990.

Unless otherwise indicated, the following terms have the following meanings:

As used herein, the term nucleoside means a glycosylamine comprising a nucleobase and a sugar. Nucleosides includes, but are not limited to, naturally occurring nucleosides, abasic nucleosides, modified nucleosides, and nucleosides having mimetic bases and/or sugar groups.

As used herein, the term nucleotide refers to a glycosomine comprising a nucleobase and a sugar having a phosphate group covalently linked to the sugar. Nucleotides may be modified with any of a variety of substituents.

As used herein, the term nucleobase refers to the base portion of a nucleoside or nucleotide. A nucleobase may comprise any atom or group of atoms capable of hydrogen bonding to a base of another nucleic acid.

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-6As used herein, the term heterocyclic base moiety refers to a nudeobase comprising a heterocycle.

As used herein, the term “deoxyribonucleotide” means a nucleotide having a hydrogen at the 2’ position of the sugar portion of the nucleotide. Deoxyribonucleotides may be modified with any of a variety of substituents.

As used herein, the term “ribonucleotide” means a nucleotide having a hydroxy at the 2’ position of the sugar portion of the nucleotide. Ribonucleotides may be modified with any of a variety of substituents.

As used herein, the term oligomeric compound refers to a polymeric structure comprising two or more sub-structures and capable of hybridizing to a region of a nucleic acid molecule. In certain embodiments, oligomeric compounds are oligonucleosides. In certain embodiments, oligomeric compounds are oligonucleotides. In certain embodiments, oligomeric compounds are antisense compounds. In certain embodiments, oligomeric compounds are antisense oligonucleotides. In certain embodiments, oligomeric compounds are short antisense compounds. In certain embodiments, oligomeric compounds are short antisense oligonucleotides. In certain embodiments, oligomeric compounds are chimeric oligonucleotides.

As used herein, the term “monomer” refers to a single unit of an oligomer. Monomers include, but are not limited to, nucleosides and nucleotides, whether naturally occuring or modified.

As used herein “oligonucleoside” refers to an oligonucleotide in which the intemucleoside linkages do not contain a phosphorus atom.

As used herein, the term oligonucleotide refers to an oligomeric compound comprising a plurality of linked nucleotides. In certain embodiment, (me or more nucleotides of an oligonucleotide is modified. In certain embodiments, an oligonucleotide comprises ribonucleic acid (RNA) or deoxyribonucleic acid (DNA). In certain embodiments, oligonucleotides are composed of naturallyand/or non-naturally-occurring nucleobases, sugars and covalent intemucleotide linkages, and may further include non-nucleic acid conjugates.

As used herein “intemucleotide linkage” refers to a covalent linkage between adjacent nucleotides.

As used herein, the term “monomeric linkage” refers to a covalent linkage between two monmers. Monomeric linkages include, but are not limited to intemucleotide linkages and intemucleoside linkages.

As used herein “naturally occuring intemucleotide linkage” refers to a 3' to 5' phosphodiester linkage.

As used herein, the term “antisense compound” refers to an oligomeric compound that is at least partially complementary to a target nucleic acid molecule to which it hybridizes. In certain embodiments, an antisense compound modulates (increases or decreases) expression of a target nucleic acid. Antisense compounds include, but are not limited to, compounds that are oligonucleotides, oligonucleosides,

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-7oligonucleotide analogs, oligonucleotide mimetics, and chimeric combinations of these. Consequently, while all antisense compounds are oligomeric compounds, not all oligomeric compounds are antisense compounds.

As used herein, the term “antisense oligonucleotide’' refers to an antisense compound that is an oligonucleotide.

As used herein, the term “parent antisense oligonucleotide” refers to an oligonucleotide 20 nucleotides in length having a deoxy gap region having ten 2’-deoxyribonucleotides, flanked by a first and a second wing region each having five 2’-O-(2-methoxyethyl) ribonucleotides (a 5-10-5 MOE gapmer) and comprising the sequence of the corresponding short antisense compound to which it is a parent.

As used herein, the term “short antisense compound” refers to an antisense compound about 8,9, 10,11,12,13,14,15 or 16 monomers in length. In certain embodiments, a short antisense compound has at least one high-affinity modification.

As used herein, the term “short antisense oligonucleotide” or refers to an antisense oligonucleotide about 8, 9,10,11, 12,13,14,15 or 16 nucleotides in length. In certain embodiments, a short antisense oligonucleotide has at least one high-affinity modification.

As used herein, the term “short gapmer” refers to a short antisense oligonucleotide having a first and a second wing region each independently 1 to 3 nucleotides in length and a gap region 2 to 14 nudeobase in length.

As used herein, the term “motif’ refers to the pattern of unmodified and modified nucleotides in a short antisense compound

As used herein, the term chimeric antisense oligomer refers to an antisense oligomeric compound, having at least one sugar, nudeobase or intemucleoside linkage that is differentially modified as compared to at least on other sugar, nudeobase or intemucleoside linkage within the same antisense oligomeric compound. The remainda of the sugars, nucleobases and intemucleoside linkages can be independently modified or unmodified, the same or different.

As used herein, the term chimeric antisense oligonucleotide refers to an antisense oligonucleotide, having at least one sugar, nudeobase or intemucleoside linkage that is differentially modified as compared to at least cm other sugar, nudeobase or intemucleoside linkage within the same antisense oligonucleotide. The remainder of the sugars, nucleobases and intemucleoside linkages can be independently modified or unmodified, the same or different.

As used herein, the term “mixed-backbone antisense oligonucleotide” refers to an antisense oligonucleotide wherein at least one intemucleoside linkage of the antisense oligonucleotide is different from at least one other intemucleotide linkage of the antisense oligonucleotide.

As used herein, the terni “target” refers to a protein, the modulation of which is desired.

As used herein, the term “target gene” refers to a gene encoding a target.

As used herein, the terms target nucleic acid and nucleic acid molecule encoding a target

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-8refer to any nucleic acid molecule the expression or activity of which is capable of being modulated by an antisense compound. Target nucleic acids include, but are not limited to, RNA (including, but not limited to pre-mRNA and mRNA or portions thereof) transcribed from DNA encoding a target, and also cDNA derived from such RNA, and miRNA. For example, the target nucleic acid can be a cellular gene (or mRNA transcribed from the gene) whose expression is associated with a particular disorder or disease state, or a nucleic acid molecule from an infectious agent.

As used herein, the term “targeting” or “targeted to” refers to the association of an antisense compound to a particular target nucleic acid molecule or a particular region of nucleotides within a target nucleic acid molecule.

As used herein, the term “5’ target site” refers to the nucleotide of a target nucleic acid which is complementary to the 5’-most nucleotide of a particular antisense compound.

As used herein, the term “3’ target site” refers to the nucleotide of a target nucleic acid which is complementary to the 3’-most nucleotide of a particular antisense compound.

As used herein, the term “target region,” refers to a portion of a target nucleic acid to which one or more antisense compounds is complementary.

As used herein, the term “target segment” refers to a smaller or sub-portions of a region within a target nucleic acid.

As used herein, the term “nucleobase complementarity” refers to a nucleobase that is capable of base pairing with another nucleobase. For example, in DNA adenine (A) is complementary to thymine (T). For example, in RNA, adenine (A) is complementary to uracil (U). In certain embodiments, complementary nucleobase refers to a nucleobase of an antisense compound that is capable of base pairing with a nucleobase of its target nucleic acid. For example, if a nucleobase at a certain position of an antisense compound is capable of hydrogen bonding with a nucleobase at a certain position of a target nucleic acid, then the position of hydrogen bonding between the oligonucleotide and the target nucleic acid is considered to be complementary at that nucleobase pair.

As used herein, the term “non-complementary nucleobase” refers to a pair of nucleobases that do not form hydrogen bonds with one another or otherwise support hybridization.

As used herein, the term “complementary” refers to the capacity of an oligomeric compound to hybridize to another oligomeric compound or nucleic acid through nucleobase complementarity. In certain embodiments, an antisense compound and its target are complementary to each other when a sufficient number of corresponding positions in each molecule are occupied by nucleobases that can bond with each other to allow stable association between the antisense compound and the target. One skilled in the art recognizes that the inclusion of mismatches is possible without eliminating the ability of the oligomeric compounds to remain in association. Therefore, described herein are antisense compounds that may comprise up to about 20% nucleotides that are mismatched (i.e., are not nucleobase complementary to the corresponding nucleotides of the target). Preferably the antisense compounds contain no more than about 15%, more preferably not more than about 10%, most preferably not more

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-9than 5% or no mismatches. The remaining nucleotides are nucleobase complementary or otherwise do not disrupt hybridization (e.g., universal bases). One of ordinary skill in the art would recognize the compounds provided herein are at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% complementary to a target nucleic acid.

As used herein, the term “mismatch” refers to a non-complementary nucleobase within a complementary oligomeric compound.

As used herein, hybridization means the pairing of complementary oligomeric compounds (e.g., an antisense compound and its target nucleic acid). While not limited to a particular mechanism, the most common mechanism of pairing involves hydrogen bonding, which may be Watson-Crick, Hoogsteen or reversed Hoogsteen hydrogen bonding, between complementary nucleoside or nucleotide bases (nucleobases). For example, the natural base adenine is nucleobase complementary to the natural nucleobases thymidine and uracil which pair through the formation of hydrogen bonds. The natural base guanine is nucleobase complementary to the natural bases cytosine and 5-methyl cytosine. Hybridization can occur under varying circumstances.

As used herein, the term “specifically hybridizes” refers to the ability of an oligomeric compound to hybridize to one nucleic acid site with greater affinity than it hybridizes to another nucleic acid site. In certain embodiments, an antisense oligonucleotide specifically hybridizes to more than one target site.

As used herein, “designing” or “designed to” refer to the process of designing an oligomeric compound that specifically hybridizes with a selected nucleic acid molecule.

As used herein, the term modulation refers to a perturbation of function or activity when compared to the level of the function or activity prior to modulation. For example, modulation includes the change, either an increase (stimulation or induction) or a decrease (inhibition or reduction) in gene expression. As further example, modulation of expression can include perturbing splice site selection of pre-mRNA processing.

As used herein, the term “expression” refers to all the functions and steps by which a gene’s coded information is converted into structures present and operating in a cell. Such structures include, but are not limited to the products of transcription and translation.

As used herein, “variant” refers to an alternative RNA transcript that can be produced from the same genomic region of DNA. Variants include, but are not limited to “pre-mRNA variants” which are transcripts produced from the same genomic DNA that differ from other transcripts produced from the same genomic DNA in either their start or stop position and contain both intronic and exonic sequence. Variants also include, but are not limited to, those with alternate splice junctions, or alternate initiation and termination codons.

As used herein, “high-affinity modified monomer” refers to a monomer having at least one modified nucleobase, intemucleoside linkage or sugar moiety, when compared to naturally occurring monomers, such that the modification increases the affinity of an antisense compound comprising the high-affinity modified monomer to its target nucleic acid. High-affinity modifications include, but are

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-10not limited to, monomers (e.g., nucleosides and nucleotides) comprising 2*-modifed sugars.

As used herein, the term “2’-modified” or “2’-substituted” means a sugar comprising substituent at the 2’ position other than H or OH. 2’-modified monomers, include, but are not limited to, BNA’s and monomers (e.g., nucleosides and nucleotides) with 2’- substituents, such as allyl, amino, azido, thio, Oallyl, O-CrCio alkyl, -OCF₃, O-(CH₂)rO-CH₃, 2’-O(CH₂)₂SCH3_> O-(CH₂)₂-O-N(R_m)(Rn), or O-CH₂¢(=0)-1^^)(1^), where each R_m and Ιζ, is, independently, H or substituted or unsubstituted Ci-Cæ alkyl. In certain embodiments, short antisense compounds comprise a 2’modified monomer that does not have the formula 2’-O(CH2)i_IH, wherein n is one to six. In certain embodiments, short antisense compounds comprise a 2’modified monomer that does not have the formula 2’-OCH₃ In certain embodiments, short antisense compounds comprise a 2’modified monomer that does not have the formula or, in the alternative, 2’-O(CH₂)2OCH₃.

As used herein, the term “bicyclic nucleic acid” or “BNA” or “bicyclic nucleoside” or “bicyclic nucleotide” refers to a nucleoside or nucleotide wherein the furanose portion of the nucleoside includes a bridge connecting two carbon atoms on the furanose ring, thereby forming a bicyclic ring system.

As used herein, unless otherwise indicated, the term “methyleneoxy BNA” alone refers to β-Dmethyleneoxy BNA.

As used herein, the term “MOE” refers to a 2’-methoxyethyl substituent.

As used herein, the term “gapmer” refers to a chimeric oligomeric compound comprising a central region (a “gap”) and a region on either side of the central region (the “wings”), wherein the gap comprises at least one modification that is different from that of each wing. Such modifications include nucleobase, monomeric linkage, and sugar modifications as well as the absence of modification (unmodified). Thus, in certain embodiments, the nucleotide linkages in each of the wings are different than the nucleotide linkages in the gap. In certain embodiments, each wing comprises nucleotides with high affinity modifications and the gap comprises nucleotides that do not comprise that modification. In certain embodiments the nucleotides in the gap and the nucleotides in the wings all comprise high affinity modifications, but the high affinity modifications in the gap are different than the high affinity modifications in the wings. In certain embodiments, the modifications in the wings are the same as one another. In certain embodiments, the modifications in the wings are different from each other. In certain embodiments, nucleotides in the gap are unmodified and nucleotides in the wings are modified. In certain embodiments, the modification(s) in each wing are the same. In certain embodiments, the modification(s) in one wing are different from the modification(s) in the other wing. In certain embodiments, short antisense compounds are gapmers having 2’-deoxynucleotides in the gap and nucleotides with highaffinity modifications in the wing.

As used herein, the term “prodrug” refers to a therapeutic agent that is prepared in an inactive form that is converted to an active form (i.e., drug) within the body or cells thereof by the action of endogenous enzymes or other chemicals and/or conditions.

As used herein, the term “pharmaceutically acceptable salts” refers to salts of active compounds

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-11that retain the desired biological activity of the active compound and do not impart undesired toxicological effects thereto.

As used herein, the term “cap structure” or “terminal cap moiety” refers to chemical modifications, which have been incorporated at either terminus of an antisense compound.

As used herein, the term “prevention” refers to delaying or forestalling the onset or development of a condition or disease for a period of time from hours to days, preferably weeks to months.

As used herein, the term “amelioration” refers to a lessening of at least one indicator of the severity of a condition or disease. The severity of indicators may be determined by subjective or objective measures which are known to those skilled in the art.

As used herein, the term “treatment” refers to administering a composition of the invention to effect an alteration or improvement of the disease or condition. Prevention, amelioration, and/or treatment may require administration of multiple doses at regular intervals, or prior to onset of the disease or condition to alter the course of the disease or condition. Moreover, a single agent may be used in a single individual for each prevention, amelioration, and treatment of a condition or disease sequentially, or concurrently.

As used herein, the term “pharmaceutical agent” refers to a substance provides a therapeutic benefit when administered to a subject

As used herein, the term “therapeutically effective amount” refers to an amount of a pharmaceutical agent that provides a therapeutic benefit to an animal.

As used herein, “administering” means providing a pharmaceutical agent to an animal, and includes, but is not limited to administering by a medical professional and self-administering.

As used herein, the term “co-administration” refers to administration of two or more pharmaceutical agents to an animal. The two or more pharmaceutical agents may be in a single pharmaceutical composition, or may be in separate pharmaceutical compositions. Each of the two or more pharmaceutical agents may be administered through the same or different routes of administration. Co-administration encompasses administration in parallel or sequentially.

As used herein, the term “pharmaceutical composition” refers to a mixture of substances suitable for administering to an individual. For example, a pharmaceutical composition may comprise an antisense oligonucleotide and a sterile aqueous solution.

As used herein, the term “individual” refers to a human or non-human animal selected for treatment or therapy.

As used herein, the term “animal” refers to a human or non-human animal, including, but not limited to, mice, rats, rabbits, dogs, cats, pigs, and non-human primates, including, but not limited to, monkeys and chimpanzees.

As used herein, the term “subject” refers to an animal, including, but not limited to a human, to whom a pharmaceutical composition is administered.

As used herein, the term “duration” refers to the period of time during which an activity or

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-12event continues. In certain embodiments, the duration of treatment is the period of time during which doses of a pharmaceutical agent are administered.

As used herein, the term “parenteral administration,” refers to administration through injection or infusion. Parenteral administration includes, but is not limited to, subcutaneous administration, intravenous administration, or intramuscular administration.

As used herein, the term “subcutaneous administration” refers to administration just below the skin. “Intravenous administration” means administration into a vein.

As used herein, the term “dose” refers to a specified quantity of a pharmaceutical agent provided in a single administration. In certain embodiments, a dose may be administered in two or more boluses, tablets, or injections. For example, in certain embodiments, where subcutaneous administration is desired, the desired dose requires a volume not easily accommodated by a single injection. In such embodiments, two or more injections may be used to achieve the desired dose. In certain embodiments, a dose may be administered in two or more injections to minimize injection site reaction in an individual.

As used herein, the term “dosage unit” refers to a form in which a pharmaceutical agent is provided. In certain embodiments, a dosage unit is a vial comprising lyophilized antisense oligonucleotide. In certain embodiments, a dosage unit is a vial comprising reconstituted antisense oligonucleotide.

As used herein, the term “pharmaceutical agent” refers to a substance provides a therapeutic benefit when administered to an individual. For example, in certain embodiments, an antisense oligonucleotide is a pharmaceutical agent.

As used herein, the term “active pharmaceutical ingredient” refers to the substance in a pharmaceutical composition that provides a desired effect.

As used herein, the term “therapeutically effective amount” refers to an amount of a pharmaceutical agent that provides a therapeutic benefit to an individual. In certain embodiments, a therapeutically effective amount of an antisense compound is the amount that needs to be administered to result in an observable benefit.

As used herein, the term “hypercholesterolemia” refers to a condition characterized by elevated serum cholesterol.

As used herein, the term “hyperlipidemia” refers to a condition characterized by elevated serum lipids.

As used herein, the term “hypertriglyceridemia” refers to a condition characterized by elevated triglyceride levels.

As used herein, the term “non-familial hypercholesterolemia” refers to a condition characterized by elevated cholesterol that is not the result of a single inherited gene mutation.

As used herein, the term “polygenic hypercholesterolemia” refers to a condition characterized by elevated cholesterol that results from the influence of a variety of genetic factors. In certain embodiments, polygenic hypercholesterolemia may be exacerbated by dietary intake of lipids.

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-13As used herein, the term “familial hypercholesterolemia (FH)” refers to an autosomal dominant metabolic disorder characterized by a mutation in the LDL-receptor (LDL-R) gene, markedly elevated LDL-C and premature onset of atherosclerosis. A diagnosis of familial hypercholesterolemia is made when a individual meets one or more of the following criteria* genetic testing confirming 2 mutated LDLreceptor genes; genetic testing confirming one mutated LDL-receptor gene; document history of untreated serum LDL-cholesterol greater than 500 mg/dL; tendinous and/or cutaneous xanthoma prior to age 10 years; or, both parents have documented elevated serum LDL-cholesterol prior to lipid-lowering therapy consistent with heterozygous familial hypercholesterolemia

As used herein, the term “homozygous familial hypercholesterolemia” or “HoFH” refers to a condition characterized by a mutation in both maternal and paternal LDL-R genes.

As used herein, the term “heterozygous familial hypercholesterolemia” or “HeFH” refers to a condition characterized by a mutation in either the maternal or paternal LDL-R gene.

As used herein, the term “mixed dyslipidemia” refers to a condition characterized by elevated serum cholesterol and elevated serum triglycerides.

As used herein, the term “diabetic dyslipidemia” or “Type II diabetes with dyslipidemia” refers to a condition characterized by Type II diabetes, reduced IIDL-C, elevated serum triglycerides, and elevated small, dense LDL particles.

As used herein, the term “CHD risk equivalents,” refers to indicators of clinical atherosclerotic disease that confer a high risk for coronary heart disease. For example, in certain embodiments, CHD risk equivalents include, without limitation, clinical coronary heart disease, symptomatic carotid artery disease, peripheral arterial disease, and/or abdominal aortic aneurysm.

As used herein, the term “non-alcoholic fatty liver disease (NAFLD)” refers to a condition characterized by fatty inflammation of the liver that is not due to excessive alcohol use (for example, alcohol consumption of over 20 g/day). In certain embodiments, NAFLD is related to insulin resistance and the metabolic syndrome.

As used herein, the term “non-alcoholic steatohepatitis (NASH)” refers to a condition characterized by inflammation and the accumulation of fat and fibrous tissue in the liver, that is not due to excessive alcohol use. NASH is an extreme form of NAFLD.

As used herein, the term “major risk factors” refers to factors that contribute to a high risk for a particular disease or condition. In certain embodiments, major risk factors for coronary heart disease include, without limitation, cigarette smoking, hypertension, low HDL-C, family history of coronary heart disease, and age.

As used herein, the term “CHD risk factors” refers to CHD risk equivalents and major risk factors.

As used herein, the term “coronary heart disease (CHD)” refers to a narrowing of the small blood vessels that supply blood and oxygen to the heart, which is often a result of atherosclerosis.

As used herein, the term “reduced coronary heart disease risk” refers to a reduction in the

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-14likelihood that a individual will develop coronary heart disease. In certain embodiments, a reduction in coronary heart disease risk is measured by an improvement in one or more CHD risk factors, for example, a decrease in LDL-C levels.

As used herein, the term “atherosclerosis” refers to a hardening of the arteries affecting large and medium-sized arteries and is characterized by the presence of fatty deposits. The fatty deposits are called atheromas or “plaques,” which consist mainly of cholesterol and other fats, calcium and scar tissue, and damage the lining of arteries.

As used herein, the term “history of coronary heart disease” refers to the occurrence of clinically evident coronary heart disease in the medical history of a individual or a individual’s family member.

As used herein, the term “Early onset coronary heart disease” refers to a diagnosis of coronary heart disease prior to age 50.

As used herein, the term “statin intolerant individual” refers to a individual who as a result of statin therapy experiences one or more of creatine kinase increases, liver function test abnormalities, muscle aches, or central nervous system side effects.

As used herein, the term “efficacy” refers to the ability to produce a desired effect. For example, efficacy of a lipid-lowering therapy may be reduction in the concentration of one or more of LDL-C, VLDL-C, IDL-C, non-HDL-C, ApoB, lipoprotein(a), or triglycerides.

As used herein, the term “acceptable safety profile” refers to a pattern of side effects that is within clinically acceptable limits.

As used herein, the term “side effects” refers to physiological responses attributable to a treatment other than desired effects. In certain embodiments, side effects include, without limitation, injection site reactions, liver function test abnormalities, renal function abnormalities, liver toxicity, renal toxicity, central nervous system abnormalities, and myopathies. For example, increased aminotransferase levels in serum may indicate liver toxicity or liver function abnormality. For example, increased bilirubin may indicate liver toxicity or liver function abnormality.

As used herein, the term “injection site reaction” refers to inflammation or abnormal redness of skin at a site of injection in an individual.

As used herein, the term “individual compliance” refers to adherence to a recommended or prescribed therapy by an individual.

As used herein, the term “lipid-lowering therapy” refers to a therapeutic regimen provided to a individual to reduce one or more lipids in a individual. In certain embodiments, a lipid-lowering therapy is provide to reduce one or more of ApoB, total cholesterol, LDL-C, VLDL-C, IDL-C, non-HDI^C, triglycerides, small dense LDL particles, and Lp(a) in an individual.

As used herein, the term “lipid-lowering agent” refers to a pharmaceutical agent provided to a individual to achieve a lowering of lipids in the individual. For example, in certain embodiments, a lipidlowering agent is provided to an individual to reduce one or more of ApoB, LDL-C, total cholesterol, and triglycerides.

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-15As used herein, the term “LDL-C target” refers to an LDL-C level that is desired following lipid-lowering therapy.

As used herein, the term “comply” refers to the adherence with a recommended therapy by an individual.

As used herein, the term “recommended therapy” refers to a therapeutic regimen recommended by a medical professional for the treatment, amelioration, or prevention of a disease.

As used herein, the term “low LDL-receptor activity” refers to LDL-receptor activity that is not sufficiently high to maintain clinically acceptable levels of LDL-C in the bloodstream.

As used herein, die term “cardiovascular outcome” refers to the occurrence of major adverse cardiovascular events.

As used herein, the term “improved cardiovascular outcome” refers to a reduction in the occurrence of major adverse cardiovascular events, or the risk thereof. Examples of major adverse cardiovascular events include, without limitation, death, reinfarction, stroke, cardiogenic shock, pulmonary edema, cardiac arrest, and atrial dysrhythmia.

As used herein, the term “surrogate markers of cardiovascular outcome” refers to indirect indicators of cardiovascular events, or the risk thereof. For example, surrogate markers of cardiovascular outcome include carotid intimal media thickness (CIMT). Another example of a surrogate marker of cardiovascular outcome includes atheroma size. Atheroma size may be determined by intravascular ultrasound (IVUS).

As used herein, the term “increased HDL-C” refers to an increase in serum HDL-C in an individual over time.

As used herein, the term “lipid-lowering” refers to a reduction in one or more serum lipids in an individual over time.

As used herein, the term “metabolic disorder” refers to a condition characterized by an alteration or disturbance in metabolic function. “Metabolic” and “metabolism” are teims well know in the art and generally include the whole range of biochemical processes that occur within a living organism. Metabolic disorders include, but are not limited to, hyperglycemia, prediabetes, diabetes (type I and type II), obesity, insulin resistance and metabolic syndrome.

As used herein, the terni “metabolic syndrome” refers to a clustering of lipid and non-lipid cardiovascular risk factors of metabolic origin. It has been closely linked to the generalized metabolic disorder known as insulin resistance. The National Cholesterol Education Program (NCEP) Adult Treatment Panel III (ΑΤΡΠΙ) established criteria for diagnosis of metabolic syndrome when three or more of five risk determinants are present. The five risk determinants are abdominal obesity defined as waist circumference of greater than 102 cm for men or greater than 88cm for women, triglyceride levels greater than or equal to 150 mg/dL, HDL cholesterol levels of less than 40 mg/dL for men and less than 50 mg/dL for women, blood pressure greater than or equal to 130/85 mm Hg and fasting glucose levels greater than or equal to 110 mg/dL. These determinants can be readily measured in clinical practice

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-16(JAMA, 2001,285: 2486-2497).

The term alkyl, as used herein, refers to a saturated straight or branched hydrocarbon radical containing up to twenty four carbon atoms. Examples of alkyl groups include, but are not limited to, methyl, ethyl, propyl, butyl, isopropyl, n-hexyl, octyl, decyl, dodecyl and the like. Alkyl groups typically include from 1 to about 24 carbon atoms, more typically from 1 to about 12 carbon atoms (C|-C|₂ alkyl) with from 1 to about 6 carbon atoms being more preferred. The term lower alkyl as used herein includes from 1 to about 6 carbon atoms. Alkyl groups as used herein may optionally include one or more further substituent groups.

The term alkenyl, as used herein, refers to a straight or branched hydrocarbon chain radical containing up to twenty four carbon atoms and having at least one carbon-carbon double bond. Examples of alkenyl groups include, but are not limited to, ethenyl, propenyl, butenyl, l-methyl-2-buten1-yl, dienes such as 1,3-butadiene and the like. Alkenyl groups typically include from 2 to about 24 carbon atoms, more typically from 2 to about 12 carbon atoms with from 2 to about 6 carbon atoms being more preferred. Alkenyl groups as used herein may optionally include one or more further substituent groups.

The term alkynyl, as used herein, refers to a straight or branched hydrocarbon radical containing up to twenty four carbon atoms and having at least one carbon-carbon triple bond. Examples of alkynyl groups include, but are not limited to, ethynyl, 1-propynyl, 1-butynyl, and the like. Alkynyl groups typically include from 2 to about 24 carbon atoms, more typically from 2 to about 12 carbon atoms with from 2 to about 6 carbon atoms being more preferred. Alkynyl groups as used herein may optionally include one or more further substituted groups.

The term aminoalkyl as used herein, refers to an amino substituted alkyl radical. This term is meant to include C1-C12 alkyl groups having an amino substituent at any position and wherein the alkyl group attaches the aminoalkyl group to the parent molecule. The alkyl and/or amino portions of the aminoalkyl group can be further substituted with substituent groups.

The term aliphatic, as used herein, refers to a straight or branched hydrocarbon radical containing up to twenty four carbon atoms wherein the saturation between any two carbon atoms is a single, double or triple bond. An aliphatic group preferably contains from 1 to about 24 carbon atoms, more typically from 1 to about 12 carbon atoms with from 1 to about 6 carbon atoms being more preferred. The straight or branched chain of an aliphatic group may be interrupted with one or more heteroatoms that include nitrogen, oxygen, sulfur and phosphorus. Such aliphatic groups interrupted by heteroatoms include without limitation polyalkoxys, such as polyalkylene glycols, polyamines, and polyimines. Aliphatic groups as used herein may optionally include further substituted groups.

The term alicyclic or alicyclyl refers to a cyclic ring system wherein the ring is aliphatic. The ring system can comprise one or more rings wherein at least one ring is aliphatic. Preferred alicyclics include rings having from about 5 to about 9 carbon atoms in the ring. Alicyclic as used herein may optionally include further substituted groups.

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-17The term alkoxy, as used herein, refers to a radical formed between an alkyl group and an oxygen atom wherein the oxygen atom is used to attach the alkoxy group to a parent molecule. Examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy, isopropoxy, nbutoxy, sec-butoxy, tert-butoxy, n-pentoxy, neopentoxy, n-hexoxy and the like. Alkoxy groups as used herein may optionally include further substituted groups.

The terms halo and halogen, as used herein, refer to an atom selected from fluorine, chlorine, bromine and iodine.

The terms aryl and aromatic, as used herein, refer to a mono- or polycyclic carbocyclic ring system radicals having one or more aromatic rings. Examples of aryl groups include, but are not limited to, phenyl, naphthyl, tetrahydronaphthyl, indanyl, idenyl and the like. Preferred aryl ring systems have from about 5 to about 20 carbon atoms in one or more rings. Aryl groups as used herein may optionally include further substituted groups.

The terms aralkyl and arylalkyl,” as used herein, refer to a radical formed between an alkyl group and an aryl group wherein the alkyl group is used to attach the aralkyl group to a pared molecule. Examples include, but are not limited to, benzyl, phenethyl and the like. Aralkyl groups as used herein may optionally include further substituted groups attached to the alkyl, the aryl or both groups that form the radical group.

The term heterocyclic radical as used herein, refers to a radical mono-, or poly-cyclic ring system that includes at least one heteroatom and is unsaturated, partially saturated or folly saturated, thereby including heteroaryl groups. Heterocyclic is also meant to include fused ring systems wherein one or more of the fused rings contain at least one heteroatom and the other rings can contain one or more heteroatoms or optionally contain no heteroatoms. A heterocyclic group typically includes at least one atom selected from sulfur, nitrogen or oxygen. Examples of heterocyclic groups include, [ 1,3]dioxolane, pyrrolidinyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, piperidinyl, piperazinyl, oxazolidinyl, isoxazolidinyl, morpholinyl, thiazolidinyl, isothiazolidinyl, quinoxalinyl, pyridazinonyl, tetrahydroforyl and the like. Heterocyclic groups as used herein may optionally include further substituted groups.

The terms heteroaryl, and heteroaromatic, as used herein, refer to a radical comprising a mono- or poly-cyclic aromatic ring, ring system or fused ring system wherein at least one of the rings is aromatic and includes one or more heteroatom. Heteroaryl is also mead to include fused ring systems including systems where one or more of the fused rings contain no heteroatoms. Heteroaryl groups typically include one ring atom selected from sulfur, nitrogen or oxygen. Examples of heteroaryl groups include, but are not limited to, pyridinyl, pyrazinyl, pyrimidinyl, pyrrolyl, pyrazolyl, imidazolyl, thiazolyl, oxazolyl, isooxazolyl, thiadiazolyl, oxadiazolyL, thiophenyl, furanyl, quinolinyl, isoquinolinyl, benzimidazolyl, benzooxazolyl, quinoxalinyl, and the like. Heteroaryl radicals can be attached to a pared molecule directly or through a linking moiety such as an aliphatic group or hetero atom. Heteroaryl groups as used herein may optionally include further substituted groups.

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-18The term heteroarylalkyl,” as used herein, refers to a heteroaryl group as previously defined having an alky radical that can attach the heteroarylalkyl group to a parent molecule. Examples include, but are not limited to, pyridinylmethyl, pyrimidinylethyl, napthyridinylpropyl and the like. Heteroarylalkyl groups as used herein may optionally include further substitutent groups on one or both of the heteroaryl or alkyl portions.

The term mono or poly cyclic structure as used in the present invention includes all ring systems that are single or polycyclic having rings that are fused or linked and is meant to be inclusive of single and mixed ring systems individually selected from aliphatic, alicyclic, aryl, heteroaryl, aralkyl, arylalkyl, heterocyclic, heteroaryl, heteroaromatic, heteroarylalkyl. Such mono and poly cyclic structures can contain rings that are uniform or have varying degrees of saturation including fully saturated, partially saturated or fully unsaturated. Each ring can comprise ring atoms selected from C, N, O and S to give rise to heterocyclic rings as well as rings comprising only C ring atoms which can be present in a mixed motif such as for example benzimidazole wherein one ring has only carbon ring atoms and the fused ring has two nitrogen atoms. The mono or poly cyclic structures can be further substituted with substituent groups such as for example phthalimide which has two =0 groups attached to one of the rings. In another aspect, mono or poly cyclic structures can be attached to a parent molecule directly through a ring atom, through a substituent group or a bifunctional linking moiety.

The term acyl, as used herein, refers to a radical formed by removal of a hydroxyl group from an organic acid an d has the general formula -C(O)-X where X is typically aliphatic, alicyclic or aromatic. Examples include aliphatic carbonyls, aromatic carbonyls, aliphatic sulfonyls, aromatic sulfinyls, aliphatic sulfinyls, aromatic phosphates, aliphatic phosphates and the like. Acyl groups as used herein may optionally include further substitutent groups.

The term hydrocarbyl includes groups comprising C, O and H. Included are straight, branched and cyclic groups having any degree of saturation. Such hydrocarbyl groups can include one or more heteroatoms selected from N, O and S and can be further mono or poly substituted with one or more substituent groups.

The terms substituent and substituent group, as used herein, include groups that are typically added to other groups or parait compounds to enhance desired properties or give desired effects. Substituent groups can be protected or unprotected and can be added to one available site or to many available sites in a parent compound. Substituent groups may also be further substituted with other substituent groups and may be attached directly or via a linking group such as an alkyl or hydrocarbyl group to a parent compound. Such groups include without limitation, halogen, hydroxyl, alkyl, alkenyl, alkynyl, acyl (-Ό(Ο)Ι<_ω), carboxyl (-QOjORa,), aliphatic groups, alicyclic groups, alkoxy, substituted oxo (-Ο-Rm), aryl, aralkyl, heterocyclic, heteroaryl, heteroarylalkyl, amino (-NRbbR«), imino(=NR_w>)₎amido (-CiOJNRtbRœOr -N(R_bb)C(O)R_8a), azido (-N₃), nitro (~NO₂), cyano (-CN), carbamido (-OCÎOJNRmAx or -N^CiOjORJ, ureido (-N(Rbb)C(O)NRbbR«), thioureido (-NfR^jQSjNRH^), guanidinyl (-N(RbbK^(=NRbb)NRbbR«)> amidinyl (-C(=NRt>b)NRbbRcc or -N(Rbb)C(NRbb)R«), thiol (-SRt*),

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-19sulfinyl (-8(0)1^), sulfonyl (-S(O)₂Ra>), sulfonamidyl (-S(O)₂NRt*R« or -N(R_bb)S(O)₂R_t,_!,) and conjugate groups. Wherein each R,,, R» and R« is, independently, H, an optionally linked chemical functional group or a further substituent group with a preferred list including without limitation H, alkyl, alkenyl, alkynyl, aliphatic, alkoxy, acyl, aryl, aralkyl, heteroaryl, alicyclic, heterocyclic and heteroarylalkyl.

B. Certain Oligomeric Compounds

In certain embodiments, it is desirable to chemically modify oligomeric compounds, compared to naturally occuring oligomers, such as DNA or RNA. Certain such modifications alter the activity of the oligomeric compound. Certain such chemical modifications can alter activity by, for example: increasing affinity of an antisense compound for its target nucleic acid, increasing its resistance to one or more nucleases, and/or altering the pharmacokinetics or tissue distribution of the oligomeric compound. In certain instances, the use of chemistries that increase the affinity of an oligomeric compound for its target can allow for the use of shorter oligomeric compounds.

1. Certain monomers

In certain embodiment, oligomeric compounds comprise one or more modified monomer. In certain such embodiments, oligomeric compounds comprise one or more high affinity monomer. In certain embodiments, such high-affinity monomer is selected from monomers (e.g., nucleosides and nucleotides) comprising 2’-modifed sugars, including, but not limited to: BNA’s and monomers (e.g., nucleosides and nucleotides) with 2’- substituents such as allyl, amino, azido, thio, O-allyl, O-C,-C₁₀alkyl, -OCFj, O-(CH₂)₂-O-CH₃, 2'-O(CH₂)2SCH3, 0-(0^-0^^)(^), or O-CH₂-C(=O)-N(R_m)(R_n), where each R_a and Ro is, independently, H or substituted or unsubstituted C|-C|₀ alkyl.

In certain embodiments, the oligomeric compounds including, but no limited to short antisense compounds of the present invention, comprise one or more high affinity monomers provided that the oligomeric compound does not comprise a nucleotide comprising a 2^0(011^11, wherein n is one to six.

In certain embodiments, the oligomeric compounds including, but no limited to short antisense compounds of the present invention, comprise one or more high affinity monomer provided that the oligomeric compound does not comprise a nucleotide comprising a 2’-OCH3 or a 2’-O(CH₂)20CH3.

In certain embodiments, the oligomeric compounds including, but no limited to short antisense compounds of the present invention, comprise one or more high affinity monomer provided that the oligomeric compound does not comprise a u-L-Methyleneoxy (4’-CH₂-O-2’) BNA.

In certain embodiments, the oligomeric compounds including, but no limited to short antisense compounds of the present invention, comprise one or more high affinity monomer provided that the oligomeric compound does not comprise a β-D-Methyleneoxy (4’-CH₂O-2’) BNA.

In certain embodiments, the oligomeric compounds including, but no limited to short antisense compounds of the present invention, comprise one or more high affinity monomer provided that the oligomeric compound does not comprise a α-L-Methyleneoxy (4’-CH2-O-2’) BNA or a β-DMethyleneoxy (4’-CH₂-O-2’) BNA.

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-20a. Certain Nucleobases

The naturally occurring base portion of a nucleoside is typically a heterocyclic base. The two most common classes of such heterocyclic bases are the purines and the pyrimidines. For those nucleosides that include a pentofuranosyl sugar, a phosphate group can be linked to the 2’, 3’ or 5’ hydroxyl moiety of the sugar. In forming oligonucleotides, those phosphate groups covalently link adjacent nucleosides to one another to form a linear polymeric compound. Within oligonucleotides, the phosphate groups are commonly referred to as forming the intemucleotide backbone of the oligonucleotide. The naturally occurring linkage or backbone of RNA and of DNA is a 3’ to 5* phosphodiester linkage.

In addition to unmodified or natural nucleobases such as the purine nucleobases adenine (A) and guanine (G), and the pyrimidine nucleobases thymine (T), cytosine (C) and uracil (U), many modified nucleobases or nucleobase mimetics known to those skilled in the art are amenable with the compounds described herein. In certain embodiments, a modified nucleobase is a nucleobase that is fairly similar in structure to the parent nucleobase, such as for example a 7-deaza purine, a 5-methyl cytosine, or a G-clamp. In certain embodiments, nucleobase mimetic include more complicated structures, such as for example a tricyclic phenoxazine nucleobase mimetic. Methods for preparation of the above noted modified nucleobases are well known to those skilled in the art.

L_____Certain sugy?

Oligomeric compounds provided herein may comprise one or more monomer, including a nucleoside or nucleotide, having a modified sugar moiety. For example, the fiiranosyl sugar ring of a nucleoside can be modified in a number of ways including, but not limited to, addition of a substituent group, bridging of two non-geminal ring atoms to form a bicyclic nucleic acid (BNA).

In certain embodiments, oligomeric compounds comprise one or more monomers that is a BNA. In certain such embodiments, BNA s include, but are not limited to, (A) a-L-Methyleneoxy (4’CH₂-O-2’) BNA, (B) β-D-Methyleneoxy (4’-CH_rO-2’) BNA, (C) Ethyleneoxy (4’-(CH₂)2-O-2’) BNA, (D) Aminooxy (4’-CH₂-O-N(R>2’) BNA and (E) Oxyamino (4’-CH_rN(R)-O2’) BNA, as depicted in Figure 1.

<img file="CA2651309C_D0001.tif" />

Figure 1. Certain BNA Structures

In certain embodimnents, BNA compounds include, but are not limited to, compounds having at least one bridge between the 4' and the 2’ position of the sugar wherein each of the bridges independently

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-21comprises 1 or from 2 to 4 linked groups independently selected from -[C(R|)(R₂)]_n-, -C(R|)=C(R₂)-, -C(R|)=N-, -C(=NR|)-, -C(=O>, -C(=S)-, -O-, -Si(R,)₂-, -S(=O)„- and -N(R,)-;

wherein:

xis0, 1, or 2, nis 1,2,3, or 4;

each R| and R2 is, independently, H, a protecting group, hydroxyl, C1-C12 alkyl, substituted C|C₁₂ alkyl, C2-C12 alkenyl, substituted C₂-C|₂ alkenyl, C₂-C₁₂ alkynyl, substituted C₂-C|₂ alkynyl, C₅-C_Maryl, substituted C5-C₂0 aryl, heterocycle radical, substituted heterocycle radical, heteroaryl, substituted heteroaryl, C₅-C₇ alicyclic radical, substituted C5-C7 alicyclic radical, halogen, OJi, NJi J₂, SJ|, N3, COOJi, acyl (C(=O)-H), substituted acyl, CN, sulfonyl (S(=O)₂-J|), or sulfoxyl (S(=O)-J|); and each J| and J₂ is, independently, H, Ci-Ci₂ alkyl, substituted C|-Ci₂ alkyl, C₂-C₁₂ alkenyl, substituted C₂-C|₂ alkenyl, C₂-C|₂ alkynyl, substituted C2-C|₂ alkynyl, C₅-C₂o aryl, substituted C5-C20 aryl, acyl (C(=O)-H), substituted acyl, a heterocycle radical, a substituted heterocycle radical, C|-C|₂aminoalkyl, substituted C_t-C|₂ aminoalkyl or a protecting group.

In one embodiment, each of the bridges of the BNA compounds is, independently, -[C(Ri)(R₂)]₀> -[C(RiXR₂)]₁-O-, -C(RiR₂)-N(R|)-O- or -C(RiR₂)-O-N(R|)-. In another embodiment, each of said bridges is, independently, 4'.CH_r2·, 4^,-(CH₂)₂-2·, 4'-(CH₂)3-2’, 4'-CH₂-O-2’, 4’-(CH₂)₂-O-2', 4'-CH₂-ON(R|)-2’ and 4'-CH₂-N(R|)-O-2'- wherein each Rt is, independently, H, a protecting group or C|-C|₂alkyl.

Certain BNA’s have been prepared and disclosed in the patent literature as well as in scientific literature (Singh et al., Chem. Commun., 1998,4,455-456; Koshkin et al., Tetrahedron, 1998,54, 36073630; Wahlestedt et al., Proc. Natl. Acad. Sci. U. S. A., 2000, 97, 5633-5638; Kumar et al., Bioorg. Med. Chem. Lett., 1998,8,2219-2222; WO 94/14226; WO 2005/021570; Singh et al., J. Org. Chem., 1998, 63, 10035-10039; Examples of issued US patents and published applications that disclose BNA s include, for example, U.S. Patent Nos. 7,053,207; 6,268,490; 6,770,748; 6,794,499; 7,034,133; and 6,525,191; and U.S. Pre-Grant Publication Nos. 2004-0171570; 2004-0219565; 2004-0014959; 20030207841; 2004-0143114; and 20030082807.

Also provided herein are BNAs in which the 2'-hydroxyl group of the ribosyl sugar ring is linked to the 4' carbon atom of the sugar ring thereby forming a methyleneoxy (4’-CH₂-O-2’) linkage to form the bicyclic sugar moiety (reviewed in Elayadi et al., Curr. Opinion Invens. Drugs, 2001, 2,558561; Braasch ¢/a/., Chem. Λ'ο/., 2001, i 1-7; and Orum el o/.₍ Cwrr. Opinion Mol. Ther., 2001,3,239243; see also U.S. Patents: 6,268,490 and 6,670,461). The linkage can be a methylene (-CH_r) group bridging the 2' oxygen atom and the 4' carbon atom, for which the teim methyleneoxy (4’-CH₂-O-2’) BNA is used for the bicyclic moiety; in the case of an ethylene group in this position, the term ethyleneoxy (4’-CH₂CH₂-O-2’) BNA is used (Singh et al., Chem. Commun., 1998,4,455-456: Morita et al., Bioorganic Medicinal Chemistry, 2003, 11, 2211-2226). Methyleneoxy (4’-CH_rO-2’) BNA and other bicyclic sugar analogs display very high duplex thermal stabilities with complementary DNA and

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-22RNA (Tm = +3 to +10° C), stability towards 3'-exonucleolytic degradation and good solubility properties. Potent and nontoxic antisense oligonucleotides compriseing BNAs have been described (Wahlestedt et al., Proc. Natl. Acad. Sci. U. S. A., 2000,97,5633-5638).

An isomer of methyieneoxy (4’-CHrO-2’) BNA that has also been discussed is alpha-Lmethyleneoxy (4’-CH₂-O-2’) BNA which has been shown to have superior stability against a 3’exonuclease. The alpha-L- methyleneoxy (4’-CHrO-2’) BNA's were incorporated into antisense gapmers and chimeras that showed potent antisense activity (Frieden el al., Nucleic Acids Research, 2003,2/,6365-6372).

The synthesis and preparation of the methyleneoxy (4’-CH₂-O-2’) BNA monomers adenine, cytosine, guanine, 5-methyl-cytosine, thymine and uracil, along with their oligomerization, and nucleic acid recognition properties have been described (Koshkin et al., Tetrahedron, 1998, 54, 3607-3630). BNAs and preparation thereof are also described in WO 98/39352 and WO 99/14226.

Analogs of methyleneoxy (4’-CH₂-O-2’) BNA, phosphorothioate- methyleneoxy (4’-CH₂-O-2’) BNA and 2'-thio-BNAs, have also been prepared (Kumar et al., Bioorg. Med. Chem. Lett., 1998, 8, 2219-2222). Preparation of locked nucleoside analogs compriseing oligodeoxyribonucleotide duplexes as substrates for nucleic acid polymerases has also been described (Wcngel et al., WO 99/14226 ). Furthermore, synthesis of 2-amino-BNA, a novel comformationally restricted high-affinity oligonucleotide analog has been described in the art (Singh et al., J. Org. Chem., 1998, 63, 1003510039). In addition, 2'-Amino- and 2'-methylamino-BNA's have been prepared and the thermal stability of their duplexes with complementary RNA and DNA strands has been previously reported.

Modified sugar moieties are well known and can be used to alter, typically increase, the affinity of the antisense compound for its target and/or increase nuclease resistance. A representative list of preferred modified sugars includes but is not limited to bicyclic modified sugars (BNA's), including methyleneoxy (4’-CH₂-O-2’) BNA and ethyleneoxy (4’-(CH₂)₂-O-2’ bridge) BNA ; substituted sugars, especially 2*-substituted sugars having a 2'-F, 2'-OCHj or a 2'-O(CH₂)₂-OCHj substituent group; and 4'thio modified sugars. Sugars can also be replaced with sugar mimetic groups among others. Methods for the preparations of modified sugars are well known to those skilled in the art. Some representative patents and publications that teach the preparation of such modified sugars include, but are not limited to, U.S. Patents: 4,981,957; 5,118,800; 5,319,080; 5,359,044; 5,393,878; 5,446,137; 5,466,786; 5,514,785; 5,519,134; 5,567,811; 5,576,427; 5,591,722; 5,597,909; 5,610,300; 5,627,053; 5,639,873; 5,646,265; 5,658,873; 5,670,633; 5,792,747; 5,700,920; 6,531,584; and 6,600,032; and WO 2005/121371.

In certain embodiments, BNA’s include bicyclic nucleoside having the formula:

<img file="CA2651309C_D0002.tif" />

wherein:

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-23Bx is a heterocyclic base moiety;

T, is H or a hydroxyl protecting group;

T₂ is H, a hydroxyl protecting group or a reactive phosphorus group;

Z is C|-Ct alkyl, Q-Q alkenyl, C₂-Q alkynyl, substituted C_rC₆ alkyl, substituted C₂-C₆ alkenyl, substituted C₂-C₆ alkynyl, acyl, substituted acyl, or substituted amide.

In one embodiment, each of the substituted groups, is, independently, mono or poly substituted with optionally protected substituent groups independently selected from halogen, oxo, hydroxyl, OJ_bNJ,J₂, SJ_b Nj, OCf=X)J_b OC(=X)NJ!J₂, NJ₃C(=X)NJ,J₂ and CN, wherein each J_b J₂ and J₃ is, independently, H or C|-C₆ alkyl, and X is O, S or NJ_b

In certain such embodiments, each of the substituted groups, is, independently, mono or poly substituted with substituent groups independently selected from halogen, oxo, hydroxyl, OJ_b NJ|J₂, SJ_b- N3, OC(=X)J_b and NJ₃C(=X)NJ|J₂, wherein each J_b J₂ and J₃ is, independently, H, C|-C₆ alkyl, or substituted C|-Cî alkyl and X is O or NJ_b

In certain embodiments, the Z group is CpQ alkyl substituted with one or more X^x, wherein each X‘ is independently OJb NJ,J2, SJb N3, OC(=X)Jb OCi=X)NJ,J2, NJjC^WA or CN; wherein each Jb J2 and J3 is, independently, H or Ci-Q alkyl, and X is O, S orNJ|. In another embodiment, the Z group is Ci-Q alkyl substituted with one or more X^x, wherein each X^x is independently halo (e.g., fluoro), hydroxyl, alkoxy (e.g., CH3O-), substituted alkoxy or azido.

In certain embodiments, the Z group is -CH₂X^X, wherein X^x is OJb NJ(J2, SJb N3, OC(=X)Jb OC(=X)NJ|J2, NJ3C(=X)NJ!J2 or CN; wherein each Jb J2 and J3 is, independently, H or CrC6 alkyl, and X is 0, S or NJ|. In another embodiment, the Z group is -CH2X^x, wherein X^x is halo (e.g., fluoro), hydroxyl, alkoxy (e.g., CH3O-) or azido.

In certain such embodiments, the Z group is in the (7?)-configuration:

<img file="CA2651309C_D0003.tif" />

In certain such embodiments, the Z group is in the (^-configuration:

<img file="CA2651309C_D0004.tif" />

In certain embodiments, each Ti and T₂ is a hydroxyl protecting group. A preferred list of hydroxyl protecting groups includes benzyl, benzoyl, 2,6-dichlorobenzyl, t-butyldimethylsilyl, t-butyldiphenylsilyl, mesylate, tosylate, dimethoxytrityl (DMT), 9-phenylxanthine-9-yl (Pixyl) and 9-(pmethoxyphenyl)xanthine-9-yl (MOX). In certain embodiments, T| is a hydroxyl protecting group selected from acetyl, benzyl, t-butyldimethylsilyl, t-butyldiphenylsilyl and dimethoxytrityl wherein a

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-24more preferred hydroxyl protecting group is Tj is 4,4'-dimethoxytrityl.

In certain embodiments, T₂ is a reactive phosphorus group wherein preferred reactive phosphorus groups include diisopropylcyanoethoxy phosphoramidite and H-phosphonate. In certain embodiments Tj is 4,4'-dimethoxytrityl and T₂ is diisopropylcyanoethoxy phosphoramidite.

In certain embodiments, oligomeric compounds have at least one monomer of the formula:

<img file="CA2651309C_D0005.tif" />

or of the formula:

<img file="CA2651309C_D0006.tif" />

or of the formula:

<img file="CA2651309C_D0007.tif" />

wherein

Bx is a heterocyclic base moiety;

T₃ is H, a hydroxyl protecting group, a linked conjugate group or an intemucleoside linking group 15 attached to a nucleoside, a nucleotide, an oligonucleoside, an oligonucleotide, a monomeric subunit or an oligomeric compound;

T« is H, a hydroxyl protecting group, a linked conjugate group or an intemucleoside linking group attached to a nucleoside, a nucleotide, an oligonucleoside, an oligonucleotide, a monomeric subunit or an oligomeric compound;

wherein at least one of T₃ and T₄ is an intemucleoside linking group attached to a nucleoside, a nucleotide, an oligonucleoside, an oligonucleotide, a monomeric subunit or an oligomeric compound; and

Z is Ci-Cô alkyl, CrQ alkenyl, C₂-Cî alkynyl, substituted Ci-C* alkyl, substituted C₂-Q alkenyl, substituted C₂-C$ alkynyl, acyl, substituted acyl, or substituted amide.

In one embodiment, each of the substituted groups, is, independently, mono or poly substituted 25 with optionally protected substituent groups independently selected from halogen, oxo, hydroxyl, OJ|, NJ,J_2> SJ|, N₃, OC(=X)J|, OC(=X)NJ₁J₂, NJ₃C(=X)NJiJ₂ and CN, wherein each J_b J₂ and J₃ is, independently, H or C_rC₆ alkyl, and X is O, S or NJ|.

In one embodiment, each of the substituted groups, is, independently, mono or poly substituted with substituent groups independently selected from halogen, oxo, hydroxyl, OJ_b NJ|J₂, SJ_b N₃,

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-25OC(=X)J|, and NJjQ^XjNJJj, wherein each J_b J₂ and Jj is, independently, H or Ci-Q alkyl, and X is 0 orNJi.

In certain such embodiments, at least one Z is C|-C« alkyl or substituted Ci-Cî alkyl. In certain embodiments, each Z is, independently, C_rC₆ alkyl or substituted C|-Q alkyl. In certain embodiments, at least one Z is C|-C₆ alkyl. In certain embodiments, each Z is, independently, CpCs alkyl. In certain embodiments, at least one Z is methyl. In certain embodiments, each Z is methyl. In certain embodiments, at least one Z is ethyl. In certain embodiments, each Z is ethyl. In certain embodiments, at least one Z is substituted C|-C₆ alkyl. In certain embodiments, each Z is, independently, substituted C_rC₆ alkyl. In certain embodiments, at least one Z is substituted methyl. In certain embodiments, each Z is substituted methyl. In certain embodiments, at least one Z is substituted ethyl. In certain embodiments, each Z is substituted ethyl.

In certain embodiments, at least one substituent group is C_rC₆ alkoxy (e.g., at least one Z is C_rC« alkyl substituted with one or more Ci-Ce alkoxy). In another embodiment, each substituent group is, independently, C|-C₆ alkoxy (e.g., each Z is, independently, C|-Ce alkyl substituted with one or more C_rCe alkoxy).

In certain embodiments, at least one C_rC₆ alkoxy substituent group is CH₃O- (e.g., at least one Z is CH₃OCH₂-). In another embodiment, each Ci-C_s alkoxy substituent group is CH₃O- (e.g., each Z is CHjOCH_r).

In certain embodiments, at least one substituent group is halogen (e.g., at least one Z is Ci-Q alkyl substituted with one or more halogen). In certain embodiments, each substituent group is, independently, halogen (e.g., each Z is, independently, C|-C₆ alkyl substituted with one or more halogen). In certain embodiments, at least one halogen substituent group is fluoro (e.g., at least one Z is CH₂FCHr, CHF₂CH₂- or CFjCHr). In certain embodiments, each halo substituent group is fluoro (e.g., each Z is, independently, CH₂FCH₂-, CHF₂CH₂- or CF₃CH₂-).

In certain embodiments, at least one substituent group is hydroxyl (e.g,, at least one Z is C|-C₆alkyl substituted with one or more hydroxyl). In certain embodiments, each substituait group is, independently, hydroxyl (e.g., each Z is, independently, Ci-Ce alkyl substituted with one or more hydroxyl). In certain embodiments, at least one Z is HOCH₂-. In another embodiment, each Z is HOCH₂-.

In certain embodiments, at least one Z is CH₃-, CH₃CH₂-, CH₂OCH₃-, CH₂F- or HOCH₂-. In certain embodiments, each Z is, independently, CH₃-, CH₃CH₂-, CH₂OCH₃-, CH₂F- or HOCH₂-.

In certain embodiments, at least one Z group is C1-C4 alkyl substituted with one or more X', wherein each X^x is, independently, OJh NJJ2, SJb N3, OC(=X)Jh OC(=X)NJjJ2, NJ3C(=X)NJ!J2 or CN; wherein each Ji, J2 and J3 is, independently, H or C|-C6 alkyl, and X is 0, S or NJb In another embodiment, at least one Z group is C|-Ce alkyl substituted with one or more X^x, wherein each X^x is, independently, halo (e.g., fluoro), hydroxyl, alkoxy (e.g., CH3O-) or azido.

In certain embodiments, each Z group is, independently, Cj-Q alkyl substituted with one or more

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-26X^x, wherein each X’ is independently 0J_b NJ,J₂, SJ_b N₃₎ OC(=X)J_b OC(=X)NJ,J₂, NJ₃C(=X)NJ,J₂ or CN; wherein each J_b J₂ and J₃ is, independently, H or C|-Cé alkyl, and X is O, S or NJ|. In another embodiment, each Z group is, independently, C,-C₆ alkyl substituted with one or more X*, wherein each X* is independently halo (e.g., fluoro), hydroxyl, alkoxy (e.g., CH₃O-) or azido.

In certain embodiments, at least one Z group is -CH₂X\ wherein X* is OJ_b NJ|J₂, SJ_b N₃, OC(=X)J_b OC(=X)NJjJ₂, NJ₃C(=X)NJ|J₂ or CN; wherein each J_b J₂ and J₃ is, independently, H or C,-C₆alkyl, and X is O, S or NJ| In certain embodiments, at least one Z group is -CH₂X^x, wherein X^x is halo (e.g., fluoro), hydroxyl, alkoxy (e.g., CH₃O-) or azido.

In certain embodiments, each Z group is, independently, -CH₂X^x, wherein each X^x is, independently, OJb NJ,J21 SJb N3, OC(=X)Jb OC(=X)NJ,J2, NJ3C(=X)NJiJ2 or CN; wherein each Jb J2 and J3 is, independently, H or C,-C6 alkyl, and X is O, S or NJb In another embodiment, each Z group is, independently, -CH2X*, wherein each X^x is, independently, halo (e.g., fluoro), hydroxyl, alkoxy (e.g., CH3O-) or azido.

In certain embodiments, at least one Z is CH₃-. In another embodiment, each Z is, CH₃-.

In certain embodiments, the Z group of at least one monomer is in the (R)- configuration represented by the formula:

<img file="CA2651309C_D0008.tif" />

or the formula:

<img file="CA2651309C_D0009.tif" />

or the formula:

<img file="CA2651309C_D0010.tif" />

In certain embodiments, the Z group of each monomer of the formula is in the (R)- configuration.

In certain embodiments, the Z group of at least one monomer is in the (5)- configuration represented by the formula:

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<img file="CA2651309C_D0011.tif" />

or the formula:

<img file="CA2651309C_D0012.tif" />

or the formula:

<img file="CA2651309C_D0013.tif" />

In certain embodiments, the Z group of each monomer of the formula is in the (S)- configuration.

In certain embodiments, T₃ is H or a hydroxyl protecting group. In certain embodiments, T₄ is H or a hydroxyl protecting group. In a further embodiment T₃ is an intemucleoside linking group attached to a nucleoside, a nucleotide or a monomeric subunit. In certain embodiments, T₄ is an intemucleoside linking group attached to a nucleoside, a nucleotide or a monomeric subunit. In certain embodiments,!; is an intemucleoside linking group attached to an oligonucleoside or an oligonucleotide. In certain embodiments, T₄ is an intemucleoside linking group attached to an oligonucleoside or an oligonucleotide. In certain embodiments, T₃ is an intemucleoside linking group attached to an oligomeric compound. In certain embodiments, T₄ is an intemucleoside linking group attached to an oligomeric compound. In In certain embodiments, at least one of Tj and T₄ comprises an intemucleoside linking group selected from phosphodiester or phosphorothioate.

In certain embodiments, oligomeric compounds have at least one region of at least two contiguous monomers of the formula:

<img file="CA2651309C_D0014.tif" />

<img file="CA2651309C_D0015.tif" />

or of the formula:

<img file="CA2651309C_D0016.tif" />

In certain embodiments, the oligomeric compound comprises at least two regions of at least two contiguous monomers of the above formula. In certain embodiments, the oligomeric compound comprises a gapped oligomeric compound. In certain embodiments, the oligrneric compound comprises at least one region of from about 8 to about 14 contiguous |J-D-2’-deoxyribofijranosyl nucleosides. In certain embodiments, the oligomeric compound comprises at least one region of from about 9 to about 12 contiguous P-D-2'-deoxyribofuranosyl nucleosides.

In certain embodiments, monmcrs include sugar mimetics. In certain such embodiments, a mimetic is used in place of the sugar or sugar-intemucleoside linkage combination, and the nudeobase is maintained for hybridization to a selected target. Representative examples of a sugar mimetics include, but are not limited to, cyclohexenyl or morpholino. Representative examples of a mimetic for a sugarintemucleoside linkage combination include, but are not limited to, peptide nucleic acids (PNA) and morpholino groups linked by uncharged achiral linkages. In some instances a mimetic is used in place of the nudeobase. Representative nudeobase mimetics are well known in the art and include, but are not limited to, tricyclic phenoxazine analogs and universal bases (Berger et al., Nuc Acid Res. 2000, 28:2911-14). Methods of synthesis of sugar, nucleoside and nudeobase mimetics arc well known to those skilled in the art

3. Monomeric Linkages

Described herein are linking groups that link monomers (including, but not limited to, modified and unmodified nucleosides and nucleotides) together, thereby forming an oligomeric compound. The two main classes of linking groups are defined by the presence or absence of a phosphorus atom. Representative phosphorus containing linkages include, but arc not limited to, phosphodiesters (P=O), phosphotriesters, methylphosphonates, phosphoramidate, and phosphorothioates (P=S). Representative non-phosphorus containing linking groups include, but arc not limited to, methylenemethylimino (-CH₂N(CH₃)-O-CH₂-), thiodiester (-O-C(O)-S-), thionocarbamate (-O-C(O)(NH)-S-); siloxane (-O-Si(H)2-O); and N.N’-dimethylhydrazine (-CHj-N(CHj)-N(CH))-). Oligomeric compounds having non-phosphorus linking groups are referred to as oligonucleosides. Modified linkages, compared to natural phosphodiester linkages, can be used to alter, typically increase, nuclease resistance of the oligomeric compound. In certain embodiments, linkages having a chiral atom can be prepared a racemic mixtures, as separate enantomers. Representative chiral linkages include, but are not limited to, alkylphosphonates and phosphorothioates. Methods of preparation of phosphorous-containing and non-phosphorouscontaining linkages are well known to those skilled in the ait.

The oligomeric compounds described herein contain one or more asymmetric centers and thus

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-29give rise to enantiomers, diastereomers, and other stereoisomeric configurations that may be defined, in terms of absolute stereochemistry, as (R) or (S), a or β such as for sugar anomers, or as (D) or (L) such as for amino acids et al. Included in the antisense compounds provided herein are all such possible isomers, as well as their racemic and optically pure forms.

4, Oligomeric Compounds

In certain embodiments, provided herein are oligomeric compounds having reactive phosphorus groups useful for forming linkages including for example phosphodiester and phosphorothioate intemudeoside linkages. Methods of preparation and/or purification of precursors or oligomeric compounds are not a limitation of the compositions or methods provided herein. Methods for synthesis and purification of oligomeric compounds including DNA, RNA, oligonucleotides, oligonucleosides, and antisense compounds are well known to those skilled in the art.

Generally, oligomeric compounds comprise a plurality of monomeric subunits linked together by linking groups. Nonlimiting examples of oligomeric compounds include primers, probes, antisense compounds, antisense oligonucleotides, external guide sequence (EGS) oligonucleotides, alternate splicers, and siRNAs. As such, these compounds can be introduced in the form of single-stranded, double-stranded, circular, branched or hairpins and can contain structural elements such as internal or terminal bulges or loops. Oligomeric double-stranded compounds can be two strands hybridized to form double-stranded compounds or a single strand with sufficient self complementarity to allow for hybridization and formation of a fully or partially double-stranded compound.

In certain embodiments, the present invention provides chimeric oligomeric compounds. In certain such embodiments, chimeric oligomeric compounds are chimeric oligonucleotides. In certain such embodiments, the chimeric oligonucleotides comprise differently modified nucleotides. In certain embodiments, chimeric oligonucleotides are mixed-backbone antisense oligonucleotides.

In general a chimeric oligomeric compound will have modified nucleosides that can be in isolated positions or grouped together in regions that will define a particular motif. Any combination of modifications and/or mimetic groups can comprise a chimeric oligomeric compound as described herein.

In certain embodiments, chimeric oligomeric compounds typically comprise at least one region modified so as to confer increased resistance to nuclease degradation, increased cellular uptake, and/or increased binding affinity for the target nucleic acid. In certain embodiments, an additional region of the oligomeric compound may serve as a substrate for enzymes capable of cleaving RNArDNA or RNAtRNA hybrids. By way of example, RNase H is a cellular endonuclease that cleaves the RNA strand of an RNA:DNA duplex. Activation of RNase H, therefore, results in cleavage of the RNA target, thereby greatly enhancing the efficiency of inhibition of gene expression. Consequently, comparable results can often be obtained with shorter oligomeric compounds when chimeras are used, compared to for example phosphorothioate deoxyoligonucleotides hybridizing to the same target region. Cleavage of the RNA target can be routinely detected by gel electrophoresis and, if necessary, associated nucleic acid hybridization techniques known in the art.

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-30In certain embodiments, chimeric oligomeric compounds are gapmers. In certain embodiments, chimeric compounds are short antisense compounds. In certain embodiments, short antisense compounds are gapmers. In certain such embodiments, a mixed-backbone antisense oligomer has one type of intemucleotide linkages in one or both wings and a different type of intemucleotide linkages in the gap. In certain such embodiments, the mixed-backbone antisense oligonucleotide has phosphodiester linkages in the wings and phosphorothioate linkages in the gap. In certain embodiments in which the intemucleotide linkages in a wing is different from the intemucleotide linkages in the gap, the intemucleotide linkage bridging that wing and the gap is the same as the intemucleotide linkage in the wing. In certain embodiments in which the intemucleotide linkages in a wing is different from the intemucleotide linkages in the gap, the intemucleotide linkage bridging that wing and the gap is the same as the intemucleotide linkage in the gap.

C. Certain Short Antiseise Compounds

Disclosed herein are short antisense compounds 8 to 16, preferably 9 to 15, more preferably 9 to 14, more preferably 10 to 14 nucleotides in length. In certain embodiments, short antisense compounds are 9 to 14 nucleotides in length. In certain embodiments, short antisense compounds are 10 to 14 nucleotides in length. In certain embodiments, such short antisense compounds are short antisense oligonucleotides.

In certain embodiments, short antisense compounds canprise aie or more chemical modifications. In certain such embodiments, short antisense compounds comprise at least one modified nucleotide. In certain embodiments short antisense compounds comprise at least two or more modified nucleotides. In certain embodiments, short antisense compounds comprise at least one modified intemucleotide linkage. In certain embodiments, short antisense compounds are mixed-backbone oligonucleotides. In certain embodiments, short antisense compounds are chimeric oligonucleotides. In certain embodiments, short antisense oligonucleotides are uniformly modified. In certain embodiments, short antisense oligonucleotides comprise modifications independently selected at each nucleobase and at each linkage.

In certain embodiments, short antisense compounds are short gapmers. In certain such embodiments, short gapmers comprise at least one high affinity modification in one or more wings of the compound. In certain embodiments, short antisense compounds comprise 1 to 3 high-affinity modifications in each wing. In certain embodiments, high affinity modifications of the short antisense compounds allow for a target affinity similar to, or even greater than, the target affinity of longer antisense compounds. In certain embodiments, the high-affinity modified nucleotides are sugar modified nucleotides. Such sugar modified nucleotides include those comprising a bridge between the 4’ and 2’ position of the sugar. Exemplary high affinity sugar modifications include, but are not limited to, BNA s and other 2’-modifications such as 2’-MOE. In an alternate embodiment of the invention, the high affinity modification is not a 2’-O-(CH2)_DH (n-1 -6) sugar-modified nucleotide. In an additional alternate embodiment, the high affinity modified nucleotide is not a 2’-OCHj or a 2’-OCH2CH2OCHj nucleotide.

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-31In certain embodiments, the high-affinity modified nucleotides confer a T„ of at least I, at least 1.5, at least 2, at least 2.5, at least 3.0, at least 3.5 or at least 4.0 degrees per nucleotide. Some high-affinity nucleotide modifications are known in the art to increase toxicity. As shown herein, short antisense compounds having a limited number (generally 2 to 6) of high affinity modifications exhibit little to no increase in toxicity but retain or increase affinity for the target RNA, while also significantly reducing expression of the RNA target. Short antisense compounds of the invention may optionally comprise a conjugate group, such as, for example, cholesterol or Ci₆.

1. Certain Wings

In certain embodiments, the short antisense compounds comprise a 5’ wing and/or a 3’ wing. In such embodiments, the features of the 3’ wing and the features of the 5’ wing are selected independently. Thus, in such embodiments, the number of monomers in the 5’ wing and the number of monomers (length) in the 3’ wing may be the same or may be different; the modifications, if any, in the 5’ wing may be the same as the modifications, if any, in the 3’ wing or such modifications, if any, may be different; and the monomeric linkages in the 5’ wing and the monomeric linkages in the 3’ wing may be the same or they may be different.

In certain embodiments a wing comprises one, two or three monomers (i.e. has a length of 1,2, or 3). In certain embodiments, the monomers of a wing are modified. In certain such embodiments, the monomers of the wing are modified to increase affinity of the antisense compound for its target nucleic acid. In certain embodiments, die monomers of a wing are nucleosides or nucleotides. In certain such embodiments, the nucleosides or nucleotides of the wing comprise a 2’ modification. In certain such embodiments, the monomers (nucleosides or nucleotides) of the wing are BNA’s. In certain such embodiments, the monomers of the wing are selected from a-l.-Methylencoxy (4’-CH₂-O-2’) BNA, β-DMethyleneoxy (4’-CH₂-O-2’) BNA , Ethyleneoxy (4’-(CH₂)rO-2’) BNA , Aminooxy (4’-CH₂-O-N(R)2’) BNA and Oxyamino (4’-CH₂-N(R)-O-2’) BNA. In certain embodiments, the monomers of a wing comprise a substituent at the 2’ position selected from allyl, amino, azido, thio, O-allyl, 0-C|-Cio alkyl, OCFj, O-fCHîjrO-CHj, 2'-O(CH₂)₂SCHj, O-(CH₂)_rO-N(R_R)(R_n), and O-CH₂-C(=O)-N(R_m)(R_n), where each R_m and Rg is, independently, H or substituted or unsubstituted Ci-Cio alkyl. In certain embodiments, the monomers of a wing are 2’MOE nucleotides.

In certain embodiments, the monomeric linkages in a wing are naturally occurring intemucleotide linkages. In certain embodiments, the monomeric linkages in a wing are non-naturally occurring intemucleotide or intemucleoside linkages. In certain such embodiments, the monomeric linkages in the wing are more resistant to one or more nucleases than naturally occurring intemucleotide linkages. In certain such embodiments, the monomeric linkages in the wing are phosphorothioate linkages (P=S). In certain embodiments where a wing has more than one monomeric linkage, the monomeric linkages are the same as one another. In certain embodiments where a wing has more than one monomers linkage, the monomers linkages are different from each other.

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-32One of ordinary skill in the art will recognize that the features and modifications discussed above may be used in any combination to prepare a wing. The table below provides non-limiting examples showing how one might prepare a wing by selecting a certain number of monomers, monomeric modifications (if any), and monomeric linkages both within the wing.

<td> Length</td><td> Monomer type/ modifications</td><td> monomeric linkages within wing</td>

<td> 1</td><td> 2’MOE</td><td> None</td>

<td> 1</td><td> BNA</td><td> None</td>

<td> 1</td><td> Methyleneoxy BNA</td><td> None</td>

<td> 1</td><td> ENA</td><td> None</td>

<td> 2</td><td> 2’MOE</td><td> P=S</td>

<td> 2</td><td> BNA</td><td> P=S</td>

<td> 2</td><td> Methyleneoxy BNA</td><td> P=S</td>

<td> 2</td><td> ENA</td><td> P=S</td>

<td> 2</td><td> 2’MOE</td><td> P=O</td>

<td> 2</td><td> BNA</td><td> P=O</td>

<td> 2</td><td> Methyleneoxy BNA</td><td> P=O</td>

<td> 2</td><td> ENA</td><td> P-0</td>

<td> 3</td><td> 2’MOE</td><td> P=S</td>

<td> 3</td><td> BNA</td><td> P=S</td>

<td> 3</td><td> Methyleneoxy BNA</td><td> P=S</td>

<td> 3</td><td> ENA</td><td> P-S</td>

<td> 3</td><td> 2’MOE</td><td> P=O</td>

<td> 3</td><td> BNA</td><td> P=O</td>

<td> 3</td><td> Methyleneoxy BNA</td><td> P=O</td>

<td> 3</td><td> ENA</td><td> P=O</td>

In certain embodiments in which a wing comprises two, three or four monomers, those two, three or four monomers all comprise the same modifications, if any. In certain embodiments in which a wing comprises two, three or four monomers, one or more of those two, three or four nudeobases comprises 10 one or more modifications that is different from one or more of the modifications of one or more of the

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-33remaining monomers.

2. Certain Gaps

In certain embodiments, the short antisense compounds comprise a gap between the 5’ wing and the 3’ wing. In certain embodiments die gap comprises five, six, seven, eight, nine, ten, eleven, twelve, thirteen, or fourteen monomers. In certain embodiments, the monomers of the gap are unmodified deoxyribonucleotides. In certain embodiments, the monomers of the gap are unmodified ribonucleotides. In certain embodiments, gap modifications (if any) gap result in an antisense compound that, when bound to its target nucleic acid, supports cleavage by an RNase, including, but not limited to, RNase H.

In certain embodiments, the monomeric linkages in the gap are naturally occurring intemucleotide linkages. In certain embodiments, the monomeric linkages in the gap are non-naturally occurring linkages. In certain such embodiments, the monomeric linkages in the gap are more resistant to one or more nuclease than naturally occurring intemucleotide linkages. In certain such embodiments, the monomeric linkages in the gap are phosphorothioate linkages (P=S). In certain embodiments, the monomeric linkages in the gap are all the same as one another. In certain embodiments, the monomeric linkages within the gap are not all the same.

One of ordinary skill in the art will recognize that the features and modifications discussed above may be used in any combination to prepare a gap. The table below provides non-limiting examples showing how one might prepare a gap by selecting a certain number of monomers, monomeric modifications (if any), and monomeric linkages within the gap region.

<td> Length</td><td> Monomer type/ modifications</td><td> Monomeric linkages within gap</td>

<td> 5</td><td> DNA</td><td> P=S</td>

<td> 6</td><td> DNA</td><td> P=S</td>

<td> 7</td><td> DNA</td><td> P=S</td>

<td> 8</td><td> DNA</td><td> P=S</td>

<td> 9</td><td> DNA</td><td> P=S</td>

<td> 10</td><td> DNA</td><td> P=S</td>

<td> 11</td><td> DNA</td><td> P=S</td>

<td> 12</td><td> DNA</td><td> P=S</td>

<td> 13</td><td> DNA</td><td> P=S</td>

<td> 14</td><td> DNA</td><td> P=S</td>

<td> 6</td><td> DNA</td><td> P=O</td>

<td> 7</td><td> DNA</td><td> P=O</td>

<td> 8</td><td> DNA</td><td> P=O</td>

<td> 9</td><td> DNA</td><td> P=O</td>

<td> 10</td><td> DNA</td><td> P=O</td>

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<td> 11</td><td> DNA</td><td> P=O</td>

<td> 12</td><td> DNA</td><td> P=O</td>

<td> 8</td><td> RNA</td><td> P=S</td>

<td> 9</td><td> RNA</td><td> P=S</td>

<td> 10</td><td> RNA</td><td> P=S</td>

<td> 11</td><td> RNA</td><td> P=S</td>

<td> 12</td><td> RNA</td><td> P=S</td>

3. Certain Gapped Antisense Oligomeric Compounds

One of ordinary skill in the art will recognize that the wings and the gaps discussed above may be selected and then combined in a variety of combinations to generate gapped oligomeric compounds, including, but not limited to, gapped antisense oligomeric compounds, and gapped antisense oligonucleotides. The features (length, modifications, linkages) of the 5’ wing and the 3’ wing may be selected independently of one another. The features of the gap include at least one difference in modification compared to the features of the 5’ wing and at least one difference compared to the features of the 3’ wing (i.e., there must be at least one difference in modification between neighboring regions to distinguish those neighboring regions from one another). The features of die gap may otherwise be selected independently.

In certain embodiments, the monomeric linkages within a wing and the monomeric linkages within the gap are the same. In certain embodiments, the monomeric linkages within a wing and the monomeric linkages within the gap are different. In certain such embodiments, the monomeric linkage bridging the wing and the gap are the same as the monomeric linkages in the wing. In certain embodiments, the monomeric linkage bridging the wing and the gap are the same as the monomeric linkages in the gap. In certain embodiments, short antisense compounds have uniform linkages throughout the compound. In certain such embodiments, all of the linkages are phosphorothioate (P=S) linkages.

One of ordinary skill in the art will recognize that the 3’ wings, 5’ wings, gaps, and linkages discussed above may be used in any combination to prepare a gapmer. The table below provides nonlimiting examples showing how one might prepare a gapmer by selecting a certain 5’ wing, a gap, a 3’ wing and certain linkages bridging the gap and each wing.

<td colspan="3"> 5’ Wing</td><td> 5’Bridge</td><td colspan="3"> Gap</td><td> 3’ Bridge</td><td colspan="3"> 3’ Wing</td>

<td> Length</td><td> Monomer</td><td> Link</td><td> Link</td><td> Length</td><td> Monomer</td><td> Link</td><td> Link</td><td> Length</td><td> Monomer</td><td> Link</td>

<td> 2</td><td> MOE</td><td> P=S</td><td> P=S</td><td> 6</td><td> DNA</td><td> P=S</td><td> P=S</td><td> 2</td><td> MOE</td><td> P=S</td>

<td> 2</td><td> BNA</td><td> P=S</td><td> P=O</td><td> 8</td><td> DNA</td><td> P-0</td><td> P=S</td><td> 3</td><td> BNA</td><td> P=S</td>

<td> 1</td><td> MOE</td><td> None</td><td> p-s</td><td> 10</td><td> DNA</td><td> P=S</td><td> P=S</td><td> 1</td><td> MOE</td><td> P=S</td>

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<td> 2</td><td> MOE</td><td> P=S</td><td> P=S</td><td> 8</td><td> RNA</td><td> P=S</td><td> P=S</td><td> 2</td><td> MOE</td><td> P=S</td>

<td> 3</td><td> Methyleneoxy BNA</td><td> P=S</td><td> P=S</td><td> 8</td><td> RNA</td><td> P=S</td><td> P=S</td><td> 3</td><td> MOE</td><td> P=S</td>

<td> 3</td><td> DNA</td><td> P=O</td><td> P=O</td><td> 10</td><td> RNA</td><td> P-S</td><td> P=O</td><td> 3</td><td> 2ΌΗ</td><td> P=O</td>

<td> 2</td><td> 2-F</td><td> P=S</td><td> P=S</td><td> 5</td><td> RNA</td><td> P=S</td><td> P-S</td><td> 2</td><td> 2’-F</td><td> P=S</td>

<td> 1</td><td> MOE</td><td> P=O</td><td> P=S</td><td> 5</td><td> DNA</td><td> P=O</td><td> P=S</td><td> 4</td><td> MOE</td><td> P=S</td>

In certain embodiments, the oligomeric compounds disclosed herein may comprise from about 8 to about 16, preferably 9 to 15, more preferably 9 to 14, more preferably 10 to 14 monomers (i.e. from about 8 to about 16 linked monomers). One of ordinary skill in the art will appreciate that this comprehends antisense compounds of 8, 9, 10, 11, 12, 13, 14, 15 or 16 nucleobases. In certain embodiments, oligomeric compounds are antisense compounds.

In certain embodiments, short antisense compounds are 8 nucleobases in length.

In certain embodiments, short antisense compounds are 9 nucleobases in length.

In certain embodiments, short antisense compounds are 10 nucleobases in length.

In certain embodiments, short antisense compounds are 11 nucleobases in length.

In certain embodiments, short antisense compounds are 12 nucleobases in length.

In certain embodiments, short antisense compounds are 13 nucleobases in length.

In certain embodiments, short antisense compounds are 14 nucleobases in length.

In certain embodiments, short antisense compounds are 15 nucleobases in length.

In certain embodiments, short antisense compounds are 16 nucleobases in length.

In certain embodiments, short antisense compounds are 8 monomers in length. In certain embodiments, short antisense compounds are 9 monomers in length. In certain embodiments, short antisense compounds are 10 monomers in length. In certain embodiments, short antisense compounds are 11 monomers in length. In certain embodiments, short antisense compounds are monomers in length. In certain embodiments, short antisense compounds arc 13 monomers in length. In certain embodiments, short antisense compounds are 14 monomers in length. In certain embodiments, short antisense compounds are 15 monomers in length. In certain embodiments, short antisense compounds are 16 monomers in length. In certain embodiments, short antisense compounds comprise 9 to 15 monomers. In certain embodiments, short antisense compounds comprise 10 to 15 monomers. In certain embodiments, short antisense compounds comprise 12 to 14 monomers. In certain embodiments, short antisense compounds comprise 12 to 14 nucleotides or nucleosides.

One having skill in the art and informed by the short antisense compounds illustrated herein will be able, without undue experimentation, to identify further short antisense compounds.

In certain embodiments, short antisense compounds comprise a gap flanked by more than one wing on either or both sides. Thus, in certain embodiments, a short antisense compound comprises two or more 5’ wings and two or more 3’ wings. In certain embodiments, a short antisense compound comprises

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-36one 5' wing and two or more 3’ wings. In certain embodiments, a short antisense compound comprises one 3’ wing and two or more 5’ wings. Certain such embodiments comprise, for example, the following regions: a first 5’ wing - a bridge - a second 5’ wing - a bridge - a gap - a bridge - a second 3’ wing a bridge - a first 3’wing. In such embodiments, each region has at least one difference in modification 5 when compared to its neighboring region. Thus, in such embodiments, the second 5’ wing and the second 3’ wing each independently comprises one or more differences in modification compared to the gap and compared to the first 5’ wing and the first 3’ wing. In such embodiments, the modifications of the first 3’ wing and first 5’ wing may either or both be the same or different from the modifications of the gap, if any.

4, Certain Conjugate Groups

In one aspect, oligomeric compounds are modified by covalent attachment of one or more conjugate groups. In general, conjugate groups modify one or more properties of the attached oligomeric compound including but not limited to pharmacodynamic, pharmacokinetic, binding, absorption, cellular distribution, cellular uptake, charge and clearance. Conjugate groups are routinely used in the chemical 15 arts and are linked directly or via an optional linking moiety or linking group to a parent compound such as an oligomeric compound. A preferred list of conjugate groups includes without limitation, intercalators, reporter molecules, polyamines, polyamides, polyethylene glycols, thioethers, polyethers, cholesterols, thiocholesterols, cholic acid moieties, folate, lipids, phospholipids, biotin, phenazine, phenanthridine, anthraquinone, adamantane, acridine, fluoresceins, rhodamines, coumarins and dyes.

Preferred conjugate groups amenable to the present invention include lipid moieties such as a cholesterol moiety (Letsinger et al., Proc. Natl. Acad. Sci. USA, 1989, 86, 6553); cholic acid (Manoharan et al., Bioorg. Med. Chem. Lett., 1994, 4, 1053); a thioether, e.g., hexyl-S-tritylthiol (Manoharan et al., Ann. N.Y. Acad. Sci., 1992,660,306; Manoharan et al., Bioorg. Med. Chem. Let., 1993,3,2765); a thiocholesterol (Oberhauser et al., Nucl. Acids Res., 1992,20,533); an aliphatic chain, 25 e.g., dodecandiol or undecyl residues (Saison-Behmoaras et al., EMBO1,1991,10,111; Kabanov et al.,

FEBS Lett., 1990, 259, 327; Svinarchuk et al., Biochimie, 1993, 75, 49); a phospholipid, e.g., dihexadecyl-rac-glycerol or triethylammonium-1,2-di-0-hexadecyl-rac-glycero-3-H-phosphonate (Manoharan et al., Tetrahedron Lett., 1995, 36, 3651; Shea et al., Nucl. Acids Res., 1990,18, 3777); a polyamine or a polyethylene glycol chain (Manoharan et al., Nucleosides &amp; Nucleotides, 1995, 14,969);

adamantane acetic acid (Manoharan et al., Tetrahedron Lett., 1995,36,3651); a palmityl moiety (Mishra et al., Biochim. Biophys. Acta, 1995, 1264, 229); or an octadecylamine or hexylamino-carbonyloxycholesterol moiety (Crooke et al., J. Pharmacol. Exp. Ther., 1996,277,923).

Linking groups or bifunctional linking moieties such as those known in the art are amenable to the compounds provided herein. Linking groups are useful for attachment of chemical functional groups, 35 conjugate groups, reporter groups and other groups to selective sites in a parent compound such as for example an oligomeric compound. In general a bifiinctional linking moiety comprises a hydrocarbyl moiety having two functional groups. One of the functional groups is selected to bind to a parent

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-37molecule or compound of interest and the other is selected to bind essentially any selected group such as chemical functional group or a conjugate group. In some embodiments, the linker comprises a chain structure or an oligomer of repeating units such as ethylene glycol or amino acid units. Examples of functional groups that are routinely used in a bifunctional linking moiety include, but are not limited to, electrophiles for reacting with nucleophilic groups and nucleophiles for reacting with electrophilic groups. In some embodiments, bifunctional linking moieties include amino, hydroxyl, carboxylic acid, thiol, unsaturations (e.g., double or triple bonds), and the like. Some nonlimiting examples of bifunctional linking moieties include 8-amino-3,6-dioxaoctanoic acid (ADO), succinimidyl 4-(Nmaleimidomethyl) cyclohexane-1-carboxylate (SMCC) and 6-aminohexanoic acid (AHEX or AHA). Other linking groups include, but are not limited to, substituted C_rC|o alkyl, substituted or unsubstituted C2-C10 alkenyl or substituted or unsubstituted Cî-Ciq alkynyl, wherein a nonlimiting list of preferred substituent groups includes hydroxyl, amino, alkoxy, carboxy, benzyl, phenyl, nitro, thiol, thioalkoxy, halogen, alkyl, aryl, alkenyl and alkynyl.

5. Synthesis, Purification and Analysis

Oligomerization of modified and unmodified nucleosides and nucleotides can be routinely performed according to literature procedures for DNA (Protocols for Oligonucleotides and Analogs, Ed. Agrawal (1993), Humana Press) and/or RNA (Scaringe, Methods (2001), 23, 206-217. Gait et al., Applications of Chemically synthesized RNA in RNA: Protein Interactions, Ed. Smith (1998), 1-36. Gallo et al., Tetrahedron (2001), 57,5707-5713).

Oligomeric compounds provided herein can be conveniently and routinely made through the well-known technique of solid phase synthesis. Equipment for such synthesis is sold by several vendors including, for example, Applied Biosystems (Foster City, CA). Any other means for such synthesis known in the art may additionally or alternatively be employed. It is well known to use similar techniques to prepare oligonucleotides such as the phosphorothioates and alkylated derivatives. The invention is not limited by the method of antisense compound synthesis.

Methods of purification and analysis of oligomeric compounds are known to those skilled in the art. Analysis methods include capillary electrophoresis (CE) and electrospray-mass spectroscopy. Such synthesis and analysis methods can be performed in multi-well plates. The method of the invention is not limited by the method of oligomer purification.

D. Antisense

Antisense mechanisms are all those involving the hybridization of a compound with target nucleic acid, wherein the outcome or effect of the hybridization is either target degradation or target occupancy with concomitant stalling of the cellular machinery involving, for example, transcription or splicing.

One type of antisense mechanism involving target degradation includes an RNase H. RNase H is a cellular endonuclease which cleaves the RNA strand of an RNA:DNA duplex. It is known in the art that single-stranded antisense compounds which are “DNA-like” elicit RNAse H activity in mammalian

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-38cells. Activation of RNase H, therefore, results in cleavage of the RNA target, thereby greatly enhancing the efficiency of DNA-like oligonucleotidemediated inhibition of gene expression.

In certain embodiments, chemically-modified antisense compounds have a higher affinity fa* target RNAs than does non-modified DNA. In certain such embodiments, that higher affinity in turn provides increased potency allowing for the administration of lower doses of such compounds, reduced potential for toxicity and improvement in therapeutic index and decreased overall cost of therapy.

The present disclosure demonstrates that the incorporation of chemically-modified high-affinity nucleotides and nucleosides into antisense compounds allows for the design of short antisense compounds 8-16 nucleobases in length useful for the reduction of target RNAs and/or target proteins in cells, tissues, and animals, including, but not limited to, humans with increased potency and improved therapeutic index. Thus, in certain embodiments, provided herein are short antisense compounds comprising highaffinity nucleotide modifications useful for reducing a target RNA in vivo. Certain such short antisense compounds are effective at lower doses than previously described antisense compounds, allowing for a reduction in toxicity and cost of treatment. In addition, certain short antisense compounds have greater potential for oral dosing.

To address the need for more potent antisense compounds, provided herein are short antisense compounds (8-16, preferably 9 to 15, more preferably 9 to 14, more preferably 10 to 14 nucleotides in length) with increased activity in vivo relative to longer compounds. Certain short antisense compounds are gapmer compounds comprising high-affinity chemically-modified nucleotides on the 3’ and 5’ ends (wings) of the compound. In certain embodiments, the addition of high-affinity modified nucleotides allows antisense compounds to be active against, and specific for, their intended target RNA in vivo despite being shorter in length. Contemplated herein are short antisense compounds wherein each of the wings independently comprises 1 to 3 high-affinity modified nucleotides. In certain embodiments, the high-affinity modifications are sugar modifications. High-affinity modified nucleotides include, but are not limited to, BNA s or other 2’-modified nucleotides, such as 2’-M0E nucleotides. Also contemplated are short antisense compounds having at least one modified intemucleotide linkage, such as a phosphorothioate intemucleotide linkage. In certain embodiments, the short antisense compounds of the present invention can have all phosphorothioate intemucleoside linkages. The short antisense compounds optionally comprise a conjugate group. As shown herein, short antisense compounds have greater affinity for target RNA than they have for DNA and are significantly more potent in vivo as shown by reduction of target mRNA as well as by amelioration of a variety of disease indications.

As used herein, an RNA which is involved in regulating glucose metabolism or clearance, lipid metabolism, cholesterol metabolism or insulin metabolism is any RNA involved in the biochemical pathways that regulate these processes. Such RNAs are well known in the art. Examples of target genes include, but are not limited to, ApoB-100 (also known as APOB; Ag(x) antigen; apoB-48; apolipoprotein B; apolipoprotein B-100; apolipoprotein B 48) and GCGR (also known as glucagon receptor; GR), CRP, DGAT2, GCCR, PCSK9, PTEN, PTP1B, SGLT2, and SOD1.

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1. Modulation of Target Expression

In certain embodiments, a target is identified and antisense oligonucleotides are designed to modulate that target or its expression. In certain embodiments, designing an oligomeric compound to a target nucleic acid molecule can be a multistep process. Typically the process begins with the identification of a target protein, the activity of which is to be modulated, and then identifying the nucleic acid the expression of which yields the target protein. In certain embodiments, designing of an antisense compound results in an antisense compound that is hybridizable to the targeted nucleic acid molecule. In certain embodiments, the antisense compound is an antisense oligonucleotide or antisense oligonucleoside. In certain embodiments, an antisense compound and a target nucleic acid are complementary to one another. In certain such embodiments, an antisense compound is perfectly complementary to a target nucleic acid. In certain embodiments, an antisense compound includes one mismatch. In certain embodiments, an antisense compound includes two mismatches. In certain embodiments, an antisense compound includes three or more mismatches.

Modulation of expression of a target nucleic acid can be achieved through alteration of any number of nucleic acid functions. In certain embodiments, the functions of RNA to be modulated include, but are not limited to, translocation functions, which include, but are not limited to, translocation of the RNA to a site of protein translation, translocation of the RNA to sites within the cell which are distant from the site of RNA synthesis, and translation of protein from the RNA. RNA processing functions that can be modulated include, but are not limited to, splicing of the RNA to yield one or more RNA species, capping of the RNA, 3’ maturation of the RNA and catalytic activity or complex formation involving the RNA which may be engaged in or facilitated by the RNA Modulation of expression can result in the increased level of one or more nucleic acid species or the decreased level of one or more nucleic acid species, either temporally or by net steady state level. Thus, in one embodiment modulation of expression can mean increase or decrease in target RNA or protein levels. In another embodiment modulation of expression can mean an increase or decrease of one or more RNA splice products, or a change in the ratio of two or more splice products.

In certain embodiments, expression of a target gene is modulated using an oligomeric compound comprising from about 8 to about 16, preferably 9 to 15, more preferably 9 to 14, more preferably 10 to 14 monomers (i.e. from about 8 to about 16 linked monomers). One of ordinary skill in the art will appreciate that this comprehends methods of modulating expression of a target gene using one or more antisense compounds of 8,9,10,11,12,13,14,15 or 16 nucleobases.

In certain embodiments, methods of modulating a target gene comprises use of a short antisense compound that is 8 nucleobases in length. In certain embodiments, methods of modulating a target gene comprises use of a short antisense compound that is 9 nucleobases in length. In certain embodiments, methods of modulating a target gene comprises use of a short antisense compound that is 8 nucleobases in length. In certain embodiments, methods of modulating a target gene comprises use of a short antisense compound that is 10 nucleobases in length. In certain embodiments, methods of modulating a target gene

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-40comprises use of a short antisense compound that is 10 nucleobases in length. In certain embodiments, methods of modulating a target gene comprises use of a short antisense compound that is 11 nucleobases in length. In certain embodiments, methods of modulating a target gene comprises use of a short antisense compound that is 12 nucleobases in length. In certain embodiments, methods of modulating a target gene comprises use of a short antisense compound that is 13 nucleobases in length. In certain embodiments, methods of modulating a target gene comprises use of a short antisense compound that is 14 nucleobases in length. In certain embodiments, methods of modulating a target gene comprises use of a short antisense compound that is 15 nucleobases in length. In certain embodiments, methods of modulating a target gene comprises use of a short antisense compound that is 16 nucleobases in length.

In certain embodiments, methods of modulating expression of a target gene comprises use of a short antisense compound comprising 9 to 15 monomers. In certain embodiments, methods of modulating expression of a target gene comprises use of a short antisense compound comprising 10 to 15 monomers. In certain embodiments, methods of modulating expression of a target gene comprises use of a short antisense compound comprising 12 to 14 monomers. In certain embodiments, methods of modulating expression of a target gene comprises use of a short antisense compound comprising 12 or 14 nucleotides or nucleosides.

2. Hybridization

In certain embodiments, antisense compounds specifically hybridize when there is a sufficient degree of complementarity to avoid non-specific binding of the antisense compound to non-target nucleic acid sequences under conditions in which specific binding is desired, i.e., under physiological conditions in the case of in vivo assays or therapeutic treatment, and under conditions in which assays are performed in the case of in vitro assays.

As used herein, “stringent hybridization conditions” or “stringent conditions” refers to conditions under which an antisense compound will hybridize to its target sequence, but to a minimal number of other sequences. Stringent conditions are sequence-dependent and will be different in different circumstances, and “stringent conditions” under which antisense compounds hybridize to a target sequence are determined by the nature and composition of the antisense compounds and the assays in which they are being investigated.

3, Complementarity

It is understood in the art that incorporation of nucleotide affinity modifications may allow for a greater number of mismatches compared to an unmodified compound. Similarly, certain oligonucleotide sequences may be more tolerant to mismatches than other oligonucleotide sequences. One of ordinary skill in the art is capable of determining an appropriate number of mismatches between oligonucleotides, or between an oligonucleotide and a target nucleic acid, such as by determining melting temperature (T_m). Tn or T_m can be calculated by techniques that are familiar to one of ordinary skill in the art. For example, techniques described in Freier et al. (Nucleic Acids Research, 1997, 25, 22: 4429-4443) allow

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-41one of ordinary skill in the art to evaluate nucleotide modifications for their ability to increase the melting temperature of an RNA:DNA duplex.

4, Identity

Antisense compounds, or a portion thereof, may have a defined percent identity to a SEQ ID NO, or a compound having a specific Isis number. As used herein, a sequence is identical to the sequence disclosed herein if it has the same nucleobase pairing ability. For example, an RNA which contains uracil in place of thymidine in the disclosed sequences of the compounds described herein would be considered identical as they both pair with adenine. This identity may be over the entire length of the oligomeric compound, or in a portion of the antisense compound (e.g., nucleobases 1-20 of a 27-mer may be compared to a 20-mer to deteimine percent identity of the oligomeric compound to the SEQ ID NO. It is understood by those skilled in the art that an antisense compound need not have an identical sequence to those described herein to function similarly to the antisense compound described herein. Shortened versions of antisense compounds taught herein, or non-identical versions of the antisense compounds taught herein, are also provided herein. Non-identical versions are those wherein each base does not have the same pairing activity as the antisense compounds disclosed herein. Bases do not have the same pairing activity by being shorter or having at least one abasic site. Alternatively, a non-identical version can include at least one base replaced with a different base with different pairing activity (e.g., G can be replaced by C, A, or T). Percent identity is calculated according to the number of bases that have identical base pairing corresponding to the SEQ ID NO or antisense compound to which it is being compared. The non-identical bases may be adjacent to each other, dispersed through out the oligonucleotide, or both.

For example, a 16-mer having the same sequence as nucleobases 2-17 of a 20-mer is 80% identical to the 20-mer. Alternatively, a 20-mer containing four nucleobases not identical to the 20-mer is also 80% identical to the 20-mer. A 14-mer having the same sequence as nucleobases 1-14 of an 18-mer is 78% identical to the 18-mer. Such calculations are well within the ability of those skilled in the art.

The percent identity is based on the percent of nucleobases in the original sequence present in a portion of the modified sequence. Therefore, a 30 nucleobase antisense compound comprising the full sequence of the complement of a 20 nucleobase active target segment would have a portion of 100% identity with the complement of the 20 nucleobase active target segment, while further comprising an additional 10 nucleobase portion. In the context of the instant description, the complement of an active target segment may constitute a single portion. In preferred embodiments, the oligonucleotides provided herein are at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical to at least a portion of the complement of the active target segments presented herein.

E. Target Nucleic Acids, Regions and Segments

In certain embodiments, short antisense compounds may be designed to target any target nucleic

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-42acid. In certain embodiments, the target nucleic acid encodes a target that is clinically relevant. In such embodiments, modulation of the target nucleic acid results in clinical benefit. Certain target nucleic acids include, but are not limited to, the target nucleic acids illustrated in Table 1.

In certain embodiments, a target nucleic acid is a nucleic acid molecule encoding ApoB. Nucleic acid molecules that encode ApoB include, without limitation, SEQ ID NO: 1 and SEQ ID NO: 2.

In certain embodiments, a target nucleic acid is a nucleic acid molecule encoding SGLT2. Nucleic acid molecules that encode SGLT2 include, without limitation, SEQ ID NO: 3.

In certain embodiments, a target nucleic acid is a nucleic acid molecule encoding PCSK9.

Nucleic acid molecules that encode PCSK9 include, without limitation, SEQ ID NO: 4.

In certain embodiments, a target nucleic acid is a nucleic acid molecule encoding SOD1. Nucleic acid molecules that encode SOD1 include, without limitation, SEQ ID NO: 5.

In certain embodiments, a target nucleic acid is a nucleic acid molecule encoding CRP. Nucleic acid molecules that encode CRP include, without limitation, SEQ ID NO: 6.

In certain embodiments, a target nucleic acid is a nucleic acid molecule encoding GCCR. Nucleic acid molecules that encode GCCR include, without limitation, SEQ ID NO: 7 and SEQ ID NO: 8.

In certain embodiments, a target nucleic acid is a nucleic acid molecule encoding GCGR. Nucleic acid molecules that encode GCGR include, without limitation, SEQ ID NO: 9.

In certain embodiments, a target nucleic acid is a nucleic acid molecule encoding DGAT2. Nucleic acid molecules that encode DGAT2 include, without limitation, SEQ ID NO: 10.

In certain embodiments, a target nucleic acid is a nucleic acid molecule encoding PTP1B. Nucleic acid molecules that encode PTP1B include, without limitation, SEQ ID NO: 11 and SEQ ID NO: 12.

In certain embodiments, a target nucleic acid is a nucleic acid molecule encoding PTEN. Nucleic acid molecules that encode PTEN include, without limitation, SEQ ID NO: 14 or SEQ ID NO: 15.

Table 1: Certain Target Nucleic Acids

<td> Target</td><td> Species</td><td> GENBANK® Accession Number</td><td> SEQ ID NO</td>

<td> ApoB</td><td> Human</td><td> NM 000384.1</td><td> 1</td>

<td> ApoB</td><td> Mouse</td><td> XM 137955.5</td><td> 2</td>

<td> SGLT2</td><td> Human</td><td> NM 003041.1</td><td> 3</td>

<td> PCSK9</td><td> Human</td><td> NMJ74936.2</td><td> 4</td>

<td> SOD1</td><td> Human</td><td> X02317.1</td><td> 5</td>

<td> CRP</td><td> Human</td><td> NM J)00567.1</td><td> 6</td>

<td> GCCR</td><td> Mouse</td><td> BCO31885.1</td><td> 7</td>

<td> GCCR</td><td> Human</td><td> Nucleotides 1 to 10600 of AC012634</td><td> 8</td>

<td> GCGR</td><td> Human</td><td> NM 000160.1</td><td> 9</td>

<td> DGAT2</td><td> Human</td><td> NM 032564.2</td><td> 10</td>

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<td> PTP1B</td><td> Human</td><td> NM 002827.2</td><td> 11</td>

<td> PTP1B</td><td> Human</td><td> Nucleotides 1417800 to 1425600 of NT 011362.9</td><td> 12</td>

<td> PTEN</td><td> Mouse</td><td> U92437.1</td><td> 13</td>

<td> PTEN</td><td> Human</td><td> NM 000314.4</td><td> 14</td>

<td> PTEN</td><td> Human</td><td> Nucleotides 8063255 to 8167140 of NT 033890.3</td><td> 15</td>

The targeting process usually includes determination of at least one target region, segment, or site within the target nucleic acid for the antisense interaction to occur such that the desired effect will result.

In certain embodiments, the 5’-most nucleotide of a target region is the 5’ target site of a short antisense compound and the 3’-most nucleotide of a target region is the 3’ target site of the same short antisense compound. In certain embodiments, the 5’-most nucleotide of a target region is the 5’ target site of a short antisense compound and the 3’-most nucleotide of a target region is the 3’ target site of a different short antisense compound. In certain embodiments, a target region comprises a nucleotide sequence within 10,15, or 20 nucleotides of a 5’ target site or a 3’ target site.

In certain embodiments, a target region is a structurally defined region of the nucleic acid. For example, in certain such embodiments, a target region may encompass a 3’ UTR, a 5’ UTR, an exon, an intron, a coding region, a translation initiation region, translation termination region, or other defined nucleic acid region.

The locations on the target nucleic acid defined by having one or more active short antisense compounds targeted thereto are referred to as “active target segments.” In certain embodiments, the target nucleic acid having one or more active short antisense compounds targeted thereto is a target RNA. When an active target segment is defined by multiple short antisense compounds, the compounds are preferably separated by no more than about 10 nucleotides on the target sequence, more preferably no more than about 5 nucleotides on the target sequence, even more preferably the short antisense compounds arc contiguous, most preferably the short antisense compounds are overlapping. There may be substantial variation in activity (e.g., as defined by percent inhibition) of the short antisense compounds within an active target segment. Active short antisense compounds are those that modulate the expression of their target nucleic acid, including but not limited to a target RNA. Active short antisense compounds inhibit expression of their target RNA at least 10%, preferably 20%. In a preferred embodiment, at least about 50%, preferably about 70% of the short antisense compounds targeted to the active target segment modulate expression of their target RNA at least 40%. In a more preferred embodiment, the level of inhibition required to define an active short antisense compound is defined based on the results from the screen used to define the active target segments.

A suitable target segment is at least about an 8-nucleobase portion of a target region to which an active short antisense compound is targeted. Target segments can include DNA or RNA sequences that comprise at least the 8 consecutive nucleobases from the 5-terminus of one of the illustrative target segments (the remaining nucleobases being a consecutive stretch of the same DNA or RNA beginning

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-44immediately upstream of the 5-tennmus of the target segment and continuing until the DNA or RNA comprises about 8 to about 16 nucleobases). Target segments are also represented by DNA or RNA sequences that comprise at least the 8 consecutive nucleobases from the 3*-terminus of one of the illustrative target segments (the remaining nucleobases being a consecutive stretch of the same DNA or RNA beginning immediately downstream of the 3*-terminus of the target segment and continuing until the DNA or RNA comprises about 8 to about 16 nucleobases). It is also understood that antisense target segments may be represented by DNA or RNA sequences that comprise at least 8 consecutive nucleobases from an internal portion of the sequence of an illustrative target segment, and may extend in either or both directions until the short antisense compound comprises about 8 to about 16 nucleobases. One having skill in the art armed with the target segments illustrated herein will be able, without undue experimentation, to identify further target segments.

Once one or more target regions, segments or sites have been identified, short antisense compounds are chosen which are sufficiently complementary to the target, i.e., hybridize sufficiently well and with sufficient specificity, to give the desired effect.

The short antisense compounds may also be targeted to regions of the target nucleobase sequence comprising any consecutive nucleobases 8 to 16 nucleobases in length along the target nucleic acid molecule.

Target segments 8-16 nucleobases in length comprising a stretch of at least eight (8) consecutive nucleobases selected from within the illustrative target segments are considered to be suitable for targeting as well. Thus, the short antisense compounds may also encompass 8-16 nucleobases within those segments identified herein as beginning at a particular 5’ target site. Any segment of 8,9,10,11, or more preferably 12, 13, 14, 15 or 16 contiguous nucleobases in a 50, preferably 25, more preferably 16 nucleobase perimeter around these regions are also considered to be suitable for targeting.

In a further embodiment, the “suitable target segments” identified herein may be employed in a screen for additional short antisense compounds that modulate the expression of a target nucleic acid. “Modulators” are those compounds that decrease or increase the expression of a target nucleic acid and which comprise at least an 8-nucleobase portion which is complementary to a target segment. The screening method comprises the steps of contacting a target segment of a nucleic acid with one or more candidate modulators, and selecting for one or more candidate modulators which decrease or increase the expression of a target nucleic acid. Once it is shown that the candidate modulator or modulators are capable of modulating (e.g. either decreasing or increasing) the expression of a target nucleic acid, the modulator may then be employed in further investigative studies of the function of the target, or for use as a research, diagnostic, or therapeutic agent in accordance with the present invention.

For all short antisense compounds discussed herein, sequence, monomer, monomeric modification, and monomeric linkage may each be selected independently. In certain embodiments, short antisense compounds are described by a motif. In such embodiments, any motif may be used with any sequence, whether or not the sequence and/or the motif is specifically disclosed herein. In certain

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-45embodimcnts, short antisense compounds comprise modifications that are not amenable to description by motif (for example, short antisense compounds comprising several different modifications and/or linkages at various positions throughout the compound). Such combinations may be incorporated for any sequence, whether or not it is disclosed herein. The sequence listing accompanying this filing provides certain nucleic acid sequences independent of chemical modification. Though that listing identifies each sequence as either “RNA” or “DNA” as required, in reality, those sequences may be modified with any combination of chemical modifications and/or motifs.

In certain embodiments, short antisense compounds comprise at least one high-affinity modified monomer. In certain embodiments, provided are short antisense compounds targeted to nucleic acid molecules encoding targets including, but not limited to, ApoB-100 (also known as APOB; Ag(x) antigen; apoB-48; apolipoprotein B; apolipoprotein B-100; apolipoprotein B-48), GCGR (also known as glucagon receptor; GR), CRP, DGAT2, GCCR, PCSK.9, PTEN, PTP1B, SGLT2, and SOD1. In certain such embodiments, such short antisense compounds are targeted to a nucleic acid molecule encoding any of those targets.

F. Certain Targets

In certain embodiments, short antisense compounds may be designed to modulate any target. In certain embodiments, the target is clinically relevant. In such embodiments, modulation of the target results in clinical benefit. Certain targets are preferentially expressed in the kidney. Certain targets are preferentially expressed in the liver. Certain targets are associated with a metabolic disorder. Certain targets are associated to a cardiovascular disorder. In certain embodiments, a target is selected from: ApoB, SGLT2, PCSK9, SOD1, CRP, GCCR, GCGR, DGAT2, PTP1B, and PTEN. In certain embodiments, a target is selected from: ApoB, SGLT2, PCSK9, SOD1, CRP, GCCR, GCGR, DGAT2, and PTP1B. In certain embodiments, a target is any protein other than SGLT2.

In certain embodiments, short antisense compounds exhibit liver and kidney-specific target RNA reduction in vivo. Such property renders those short antisense compounds particularly useful for inhibition of many target RNAs involved in metabolic and cardiovascular diseases. Thus, provided herein are methods of treating cardiovascular or metabolic disorders by contacting said kidney or liver tissues with short antisense compounds targeted to RNAs associated with said disorders. Thus, also provided are methods for ameliorating any of a variety of metabolic or cardiovascular disease indications with the short antisense compounds of the present invention.

1. ApoB

ApoB (also known as apolipoprotein B-100; ApoB-100, apolipoprotein B-48; ApoB-48 and Ag(x) antigen), is a large glycoprotein that serves an indispensable role in the assembly and secretion of lipids and in the transport and receptor-mediated uptake and delivery of distinct classes of lipoproteins. ApoB performs a variety of activities, from the absorption and processing of dietary lipids to the regulation of circulating lipoprotein levels (Davidson and Shelness, Annu. Rev. Nutr., 2000,20,169-193).

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-46This latter property underlies its relevance in terms of atherosclerosis susceptibility, which is highly correlated with the ambient concentration of ApoB-containing lipoproteins (Davidson and Shelncss, Annu. Rev. Nutr., 2000,20,169-193). ApoB-100 is the major protein component of LDL-C and contains the domain required for interaction of this lipoprotein species with the LDL receptor. Elevated levels of LDL-C are a risk factor for cardiovascular disease, including atherosclerosis.

Definitions

ApoB” is the gene product or protein of which expression is to be modulated by administration of a short antisense compound.

“ApoB nucleic acid means any nucleic acid encoding ApoB. For example, in certain embodiments, a ApoB nucleic acid includes, without limitation, a DNA sequence encoding ApoB, an RNA sequence transcribed from DNA encoding ApoB, and an mRNA sequence encoding ApoB.

“ApoB mRNA” means an mRNA encoding ApoB.

ApoB Therapeutic Indications

In certain embodiments, the invention provides methods of modulating the expression of ApoB in an individual comprising administering a short antisense compound targeted to an ApoB nucleic acid. In certain embodiments, the invention provides methods of treating an individual comprising administering one or more pharmaceutical compositions comprising a short antisense compound targeted to an ApoB nucleic acid. In certain embodiments, the individual has hypercholesterolemia, non-familial hypercholesterolemia, familial hypercholesterolemia, heterozygous familial hypercholesterolemia, homozygous familial hypercholesterolemia, mixed dyslipidemia, atherosclerosis, a risk of developing atherosclerosis, coronary heart disease, a history of coronary heart disease, early onset coronary heart disease, one or more risk factors for coronary heart disease, type II diabetes, type II diabetes with dyslipidemia, dyslipidemia, hypertriglyceridemia, hyperlipidemia, hyperfattyacidemia, hepatic steatosis, non-alcoholic steatohepatitis, or non-alcoholic fatty liver disease.

Guidelines for lipid-lowering therapy were established in 2001 by Adult Treatment Panel III (ATP III) of the National Cholesterol Education Program (NCEP), and updated in 2004 (Grundy et al., Circulation, 2004, 110, 227-239). The guidelines include obtaining a complete lipoprotein profile, typically after a 9 to 12 hour fast, for determination of LDL-C, total cholesterol, and HDL-C levels. According to the most recently established guidelines, LDL-C levels of 130-159 mg/dL, 160-189 mg/dL, and greater than or equal to 190 mg/dL are considered borderline high, high, and very high, respectively. Total cholesterol levels of 200-239 and greater than or equal to 240 mg/dL are considered borderline high and high, respectively. HDL-C levels of less than 40 mg/dL are considered low.

In certain embodiments, the individual has been identified as in need of lipid-lowering therapy. In certain such embodiments, the individual has been identified as in need of lipid-lowering therapy according to the guidelines established in 2001 by Adult Treatment Panel III (ATP III) of the National

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-47Cholesterol Education Program (NCEP), and updated in 2004 (Grundy et al., Circulation, 2004,110,227239). In certain such embodiments, the individual in need of lipid-lowering therapy has LDL-C above 190 mg/dL. In certain such embodiments, the individual in need of lipid-lowering therapy has LDL-C above 160 mg/dL. In certain such embodiments, the individual in need of lipid-lowering therapy has LDL-C above 130 mg/dL. In certain such embodiments the individual in need of lipid-lowering therapy has LDLC above 100 mg/dL. In certain such embodiments the individual in need of lipid-lowering therapy should maintain LDL-C below 160 mg/dL. In certain such embodiments the individual in need of lipid-lowering therapy should maintain LDL-C below 130 mg/dL. In certain such embodiments the individual in need of lipid-lowering therapy should maintain LDL-C below 100 mg/dL. In certain such embodiments the individual should maintain LDL-C below 70 mg/dL.

In certain embodiments the invention provides methods for reducing ApoB in an individual. In certain embodiments the invention provides methods for reducing ΛροΒ-containing lipoprotein in an individual. In certain embodiments the invention provides methods for reducing LDL-C in an individual. In certain embodiments the invention provides methods for reducing VLDL-C in an individual. In certain embodiments the invention provides methods for reducing 1DL-C in an individual. In certain embodiments the invention provides methods for reducing non-HDL-C in an individual. In certain embodiments the invention provides methods for reducing Lp(a) in an individual. In certain embodiments the invention provides methods for reducing serum triglyceride in an individual. In certain embodiments the invention provides methods for reducing liver triglyceride in an individual. In certain embodiments the invention provides methods for reducing Ox-LDL-C in an individual. In certain embodiments the invention provides methods for reducing small LDL particles in an individual. In certain embodiments the invention provides methods for reducing small VLDL particles in an individual. In certain embodiments the invention provides methods for reducing phospholipids in an individual. In certain embodiments the invention provides methods for reducing oxidized phospholipids in an individual.

In certain embodiments the invention provides methods for reducing Ox-LDL-C concentration in a subject. In certain such embodiments, the reduction in ApoB, LDL-C, VLDL-C, IDL-C, total cholesterol, non-HDL-C, Lp(a), triglyerides, or Ox-LDL-C is, independently, selected from at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, and at least 100%. In certain such embodiments, the reduction in ApoB, LDL-C, VLDL-C, IDL-C, total cholesterol, non-HDL-C, Lp(a), triglyerides, or Ox-LDL-C is, independently, selected from at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, and at least 70%. In certain such embodiments, the reduction in ApoB, LDL-C, VLDL-C, IDL-C, total cholesterol, non-HDLC, Lp(a), triglyerides, or Ox-LDL-C is, independently, selected from at least 40%, at least 50%, at least 60%, and at least 70%.

In certain embodiments, the invention provides method for raising HDL-C concentration in a subject.

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-48In certain embodiments, the methods provided by the present invention do not lower HDL-C. In certain embodiments, the methods provided by the present invention do not result in accumulation of lipids in the liver. In certain embodiments, the methods provided by the present invention do not cause hepatic steatosis.

In certain embodiments, the invention provides methods for lowering ApoB concentration in a subject while reducing side effects associated with treatment. In certain such embodiments, a side effect is liver toxicity. In certain such embodiments, a side effect is abnormal liver function. In certain such embodiments, a side effect is elevated alanine aminotransferase (ALT). In certain such embodiments, a side effect is elevated aspartate aminotransferase (AST).

In certain embodiments, the invention provides methods for lowering ApoB concentration tn a subject who is not reaching target LDIrC levels as a result of lipid-lowering therapy. In certain such embodiments, a short antisense compound targeted to an ApoB nucleic acid is the only lipid-lowering agent administered to the subject. In certain such embodiments, the subject has not complied with recommended lipid-lowering therapy. In certain such embodiments, a pharmaceutical composition of the invention is co-administered with an additional different lipid-lowering therapy. In certain such embodiments, an additional lipid-lowering therapy is LDL-apheresis. In certain such embodiments, an additional lipid-lowering therapy is a statin. In certain such embodiments, an additional lipid-lowering therapy is ezetimibe.

In certain embodiments, the invention provides methods for lowering ApoB concentration in a statin-intolerant subject. In certain such embodiments, the subject has creatine kinase concentration increases as a result of statin administration. In certain such embodiments, the subject has liver function abnormalities as a result of statin administration. In certain such embodiments the subject has muscle aches as a result of statin administration. In certain such embodiments the subject has central nervous system side effects as a result of statin administration. In certain embodiments, the subject has not complied with recommended statin administration.

In certain embodiments, the invention provides methods for lowering liver triglycerides in a subject. In certain such embodiments, the subject has elevated liver triglycerides. In certain such embodiments, the subject has steatohepatitis. In certain such embodiments, the subject has steatosis. In certain such embodiments, liver triglyceride levels are measured by magnetic resonance imaging.

In certain embodiments, the invention provides methods for reducing coronary heart disease risk in a subject. In certain embodiments the invention provides methods for slowing the progression of atherosclerosis in a subject. In certain such embodiments the invention provides methods for stopping the progression of atherosclerosis in a subject In certain such embodiments the invention provides methods for reducing the size and/or prevalence of atherosclerotic plaques in a subject. In certain embodiments the methods provided reduce a subject’s risk of developing atherosclerosis.

In certain embodiments the methods provided improve the cardiovascular outcome in a subject. In certain such embodiments improved cardiovascular outcome is the reduction of the risk of developing

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-49coronary heart disease. In certain such embodiments, improved cardiovascular outcome is a reduction in the occurrence of one or more major cardiovascular events, which include, but are not limited to, death, myocardial infarction, reinfarction, stroke, cardiogenic shock, pulmonary edema, cardiac arrest, and atrial dysrhythmia. In certain such embodiments, the improved cardiovascular outcome is evidenced by improved carotid intimal media thickness. In certain such embodiments, improved carotid intimal media thickness is a decrease in thickness. In certain such embodiments, improved carotid intimal media thickness is a prevention an increase of intimal media thickness.

In certain embodiments a pharmaceutical composition comprising a short antisense compound targeted to an ApoB nucleic acid is for use in therapy. In certain embodiments, the therapy is the reduction of LDDC, ApoB, VLDDC, IDL-C, non-HDL-C, Lp(a), serum triglyceride, liver triglyceride, Ox-LDDC, small LDL particles, small VLDL, phospholipids, or oxidized phospholipids in an individual. In certain embodiments, the therapy is the treatment of hypercholesterolemia, non-familial hypercholesterolemia, familial hypercholesterolemia, heterozygous familial hypercholesterolemia, homozygous familial hypercholesterolemia, mixed dyslipidemia, atherosclerosis, a risk of developing atherosclerosis, coronary heart disease, a history of coronary heart disease, early onset coronary heart disease, one or more risk factors for coronary heart disease, type II diabetes, type II diabetes with dyslipidemia, dyslipidemia, hypertriglyceridemia, hyperlipidemia, hyperfattyacidemia, hepatic steatosis, non-alcoholic steatohepatitis, or non-alcoholic fatty liver disease. In additional embodiments, the therapy is the reduction of CHD risk. In certain the therapy is prevention of atherosclerosis. In certain embodiments, the therapy is the prevention of coronary heart disease.

In certain embodiments a pharmaceutical composition comprising a short antisense compound targeted to an ApoB nucleic acid is used for the preparation of a medicament for reducing LDL-C, ApoB, VLDL-C, IDL-C, non-HDL-C, Lp(a), serum triglyceride, liver triglyceride, Ox-LDL-C, small LDL particles, small VLDL, phospholipids, or oxidized phospholipids in an individual. In certain embodiments pharmaceutical composition comprising a short antisense compound targeted to an ApoB nucleic acid is used for the preparation of a medicament for reducing coronary heart disease risk. In certain embodiments a short antisense compound targeted to an ApoB nucleic acid is used for the preparation of a medicament for the treatment of hypercholesterolemia, non-familial hypercholesterolemia, familial hypercholesterolemia, heterozygous familial hypercholesterolemia, homozygous familial hypercholesterolemia, mixed dyslipidemia, atherosclerosis, a risk of developing atherosclerosis, coronary heart disease, a history of coronary heart disease, early onset coronary heart disease, one or more risk factors for coronary heart disease, type II diabetes, type Π diabetes with dyslipidemia, dyslipidemia, hypertriglyceridemia, hyperlipidemia, hyperfattyacidemia, hepatic steatosis, non-alcoholic steatohepatitis, or non-alcoholic fatty liver disease.

ApoB Combination Therapies

In certain embodiments, one or more pharmaceutical compositions comprising a short antisense

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-50compound targeted to an ApoB nucleic acid are co-administered with one or more other pharmaceutical agents. In certain embodiments, such one or more other pharmaceutical agents are designed to treat the same disease or condition as the one or more pharmaceutical compositions of the present invention. In certain such embodiments, the one or more pharmaceutical agents are lipid-lowering agents. In certain embodiments, such one or more other pharmaceutical agents are designed to treat a different disease or condition as the one or more pharmaceutical compositions of the present invention. In certain embodiments, such one or more other pharmaceutical agents are designed to treat an undesired effect of one or more pharmaceutical compositions of the present invention. In certain embodiments, one or more pharmaceutical compositions of the present invention are co-administered with another pharmaceutical agent to treat an undesired effect of that other pharmaceutical agent. In certain embodiments, one or more pharmaceutical compositions of the present invention and one or more other pharmaceutical agents are administered at the same time. In certain embodiments, one or more pharmaceutical compositions of the present invention and one or more other pharmaceutical agents are administered at different times. In certain embodiments, one or more pharmaceutical compositions of the present invention and one or more other pharmaceutical agents are prepared together in a single formulation. In certain embodiments, one or more pharmaceutical compositions of the present invention and one or more other pharmaceutical agents are prepared separately.

In certain embodiments, pharmaceutical agents that may be co-administered with a pharmaceutical composition comprising a short antisense compound targeted to an ApoB nucleic acid include lipid-lowering agents. In certain such embodiments, pharmaceutical agents that may be coadministered with a pharmaceutical composition of the present invention include, but are not limited to atorvastatin, simvastatin, rosuvastatin, and ezetimibe. In certain such embodiments, the lipid-lowering agent is administered prior to administration of a pharmaceutical composition of die present invention. In certain such embodiments, the lipid-lowering agent is administered following administration of a pharmaceutical composition of the present invention. In certain such embodiments the lipid-lowering agent is administered at the same time as a pharmaceutical composition of the present invention. In certain such embodiments the dose of a co-administered lipid-lowering agent is the same as the dose that would be administered if the lipid-lowering agent was administered alone. In certain such embodiments the dose of a co-administered lipid-lowering agent is lower than the dose that would be administered if the lipid-lowering agent was administered alone. In certain such embodiments the dose of a coadministered lipid-lowering agent is greater than the dose that would be administered if the lipid-lowering agent was administered alone.

In certain embodiments, a co-administered lipid-lowering agent is a HMG-CoA reductase inhibitor. In certain such embodiments the HMG-CoA reductase inhibitor is a statin. In certain such embodiments the statin is selected from atorvastatin, simvastatin, pravastatin, fluvastatin, and rosuvastatin.

In certain embodiments, a co-administered lipid-lowering agent is a cholesterol absorption

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-51inhibitor. In certain such embodiments, cholesterol absorption inhibitor is ezetimibe.

In certain embodiments, a co administered lipid-lowering agent is a co-formulated HMG-CoA reductase inhibitor and cholesterol absorption inhibitor. In certain such embodiments the co-formulated lipid-lowering agent is ezetimibe/simvastatin.

In certain embodiments, a co-administered lipid-lowering agent is a microsomal triglyceride transfer protein inhibitor (MTP inhibitor).

In certain embodiments, a co-administered pharmaceutical agent is a bile acid séquestrant. In certain such embodiments, the bile acid séquestrant is selected from cholestyramine, colestipol, and colesevelam.

In certain embodiments, a co-administered pharmaceutical agent is a nicotinic acid. In certain such embodiments, the nicotinic acid is selected from immediate release nicotinic acid, extended release nicotinic acid, and sustained release nicotinic acid.

In certain embodiments, a co-administered pharmaceutical agent is a fibric acid. In certain such embodiments, a fibric acid is selected from gemfibrozil, fenofibrate, clofibrate, bezafibrate, and ciprofibrate.

Further examples of pharmaceutical agents that may be co-administered with a pharmaceutical composition comprising a short antisense compound targeted to an ApoB nucleic acid include, but are not limited to, corticosteroids, including but not limited to prednisone; immunoglobulins, including, but not limited to intravenous immunoglobulin (IVlg); analgesics (e.g., acetaminophen); anti-inflammatory agents, including, but not limited to non-steroidal anti-inflammatory drugs (e.g., ibuprofen, COX-1 inhibitors, and COX-2, inhibitors); salicylates; antibiotics; antivirals; antifungal agents; antidiabetic agents (e.g., biguanides, glucosidase inhibitors, insulins, sulfonylureas, and thiazolidenediones); adrenergic modifiers; diuretics; hormones (e.g., anabolic steroids, androgen, estrogen, calcitonin, progestin, somatostan, and thyroid hormones); inununomodulators; muscle relaxants; antihistamines; osteoporosis agents (e.g., biphosphonates, calcitonin, and estrogens); prostaglandins, antineoplastic agents; psychotherapeutic agents; sedatives; poison oak or poison sumac products; antibodies; and vaccines.

In certain embodiments, a pharmaceutical composition comprising a short antisense compound targeted to an ApoB nucleic acid may be administered in conjunction with a lipid-lowering therapy. In certain such embodiments, a lipid-lowering therapy is therapeutic lifestyle change. In certain such embodiments, a lipid-lowering therapy is LDL apheresis.

In one embodiment, the antisense compounds provided herein can be used to lower the level of apolipoprotein B-containing lipoproteins in a human subject. As used herein, “apolipoprotein Bcontaining lipoprotein” refers to any lipoprotein that has apolipoprotein B as its protein component, and is understood to include LDL, VLDL, IDL, and lipoprotein(a). LDL, VLDL, IDL and lipoprotein(a) each contain one molecule of apolipoprotein B, thus a serum apolipoprotein B measurement reflects the total number of these lipoproteins. As is known in the art, each of the aforementioned lipoproteins is

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-52atherogenic. Thus, lowering one or more apolipoprotein B-containing lipoproteins in senim may provide a therapeutic benefit to a human subject. Small LDL particles are considered to be particularly atherogenic relative to large LDL particles, thus lowering small LDL particles can provide a therapeutic benefit to a human subject. Additional lipid parameters can also be determined in a subject. Reduction of total cholesterokHDL ratio or LDL:HDL ratio is a clinically desirable improvement in cholesterol ratio. Similarly, it is clinically desirable to reduce serum triglycerides in humans who exhibit elevated lipid levels.

Other indications of cardiovascular disease that can be measured in a subject include serum LDL particle size; serum LDL cholesteryl ester concentration; serum LDL cholesteryl ester composition; the extent of polyunsaturation of serum LDL cholesteryl esters; and serum HDL cholesterol levels. As used herein, “serum LDL particle size” refers to the classification of serum LDL particle size, which may be very small, small, medium, or large, and is typically expressed in g/pmol. In the context of the present invention, “serum LDL cholesteryl ester concentration” means the amount of cholesteryl ester present in LDL particles, and is typically measured as mg/dL. In the context of the present invention, “serum LDL cholesteryl ester composition” is a measurement of the percentage of saturated, monounsaturated and polyunsaturated cholesteryl ester fatty acids present in serum LDL particles. “Polyunsaturation of serum LDL cholesteryl esters” means the percentage of polyunsaturated cholesteryl ester fatty acids in serum LDL particles.

Methods of obtaining serum or plasma samples for analysis and methods of preparation of the serum samples to allow for analysis are well known to those skilled in the art. With regard to measurements of lipoproteins, cholesterol, triglyceride and cholesteryl esters, the terms “serum” and “plasma” are herein used interchangeably.

In another embodiment, the antisense compounds provided herein can be used to treat metabolic disorders. A variety of biomarkers can be used for evaluating metabolic disease. For example, blood glucose levels can be determined by a physician or even by the patient using a commonly available test kit or glucometer (for example, the Ascensia ELITE™ kit, Ascensia (Bayer), Tanytown NY, or Accucheck, Roche Diagnostics). Glycated hemoglobin (HbA_IC) can also be measured. HbA|_C is a stable minor hemoglobin variant formed in vivo via posttranslational modification by glucose, and it contains predominantly glycated NHj-terminal β-chains. There is a strong correlation between levels of HbA_lcand the average blood glucose levels over the previous 3 months. Thus HbA|_C is often viewed as the gold standard for measuring sustained blood glucose control (Bunn, H.F. et al., 1978, Science. 200, 21-7). HbAic can be measured by ion-exchange HPLC or immunoassay; home blood collection and mailing kits for HbAi_c measurement are now widely available. Serum fructosamine is another measure of stable glucose control and can be measured by a colorimetric method (Cobas Integra, Roche Diagnostics).

Certain Short Antisense Compounds Targeted to an ApoB Nucleic Add

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-53In certain embodiments, short antisense compounds are targeted to an ApoB nucleic acid having the sequence of GENBANK® Accession No. NM_000384.1, incorporated herein as SEQ ID NO: 1. In certain such embodiments, a short antisense compound targeted to SEQ ID NO: 1 is at least 90% complementary to SEQ ID NO: 1. In certain such embodiments, a short antisense compound targeted to SEQ ID NO: 1 is at least 95% complementary to SEQ ID NO: 1. In certain such embodiments, a short antisense compound targeted to SEQ ID NO: 1 is 100% complementary to SEQ ID NO: 1. In certain embodiments, a short antisense compound targeted to SEQ ID NO: 1 comprises a nucleotide sequence selected from the nucleotide sequences set forth in Table 2 and Table 3.

The nucleotide sequence set forth in each SEQ ID NO in Tables 2 and 3 is independent of any modification to a sugar moiety, a monomeric linkage, or a nucleobase. As such, short antisense compounds defined by a SEQ ID NO may comprise, independently, one or more modifications to a sugar moiety, an intemucleoside linkage, or a nucleobase. Antisense compounds described by Isis Number (Isis NO.) indicate a combination of nucleobase sequence and one or more modifications to a sugar moiety, an intemucleoside linkage, or a nucleobase.

Tables 2 and 3 illustrate examples of short antisense compounds targeted to SEQ ID NO: 1. Table 2 illustrates short antisense compounds that are 100% complementary to SEQ ID NO: 1. Table 3 illustrates short antisense compounds that have one or two mismatches with respect to SEQ ID NO: 1. The column labeled ‘gapmer motif indicates the wing-gap-wing motif of each short antisense compounds. The gap segment comprises 2’-deoxynucleotides and each nucleotide of each wing segment comprises a 2’-modified sugar. The particular 2’-modified sugar is also indicated in the ‘gapmer motif column. For example, ‘2-10-2 MOE’ means a 2-10-2 gapmer motif, where a gap segment of ten 2’deoxynucleotides is flanked by wing segments of two nucleotides, where the nucleotides of the wing segments are 2’-MOE nucleotides. Intemucleoside linkages are phosphorothioate. The short antisense compounds comprise 5-methylcytidine in place of unmodified cytosine, unless “unmodified cytosine” is listed in the gapmer motif column, in which case the indicated cytosines are unmodified cytosines. For example, “5-mC in gap only” indicates that the gap segment has 5-methylcytosines, while the wing segments have unmodified cytosines.

Table 2: Short Antisense Compounds targeted to SEQ Π) NO: 1

<td> ISIS No</td><td> 5’ Target Site</td><td> 3' Target Site</td><td> Sequence (5’-3’)</td><td> Gapmer Motif</td><td> SEQ ID NO</td>

<td> 372816</td><td> 263</td><td> 278</td><td> CCGGAGGTGCTTGAAT</td><td> 3-10-3 MOE</td><td> 16</td>

<td> 372894</td><td> 264</td><td> 277</td><td> CGGAGGTGCTTGAA</td><td> 2-10-2 MOE</td><td> 17</td>

<td> 372817</td><td> 428</td><td> 443</td><td> GAAGCCATACACCTCT</td><td> 3-10-3 MOE</td><td> 18</td>

<td> 372895</td><td> 429</td><td> 442</td><td> AAGCCATACACCTC</td><td> 2-10-2 MOE</td><td> 19</td>

<td> 372818</td><td> 431</td><td> 446</td><td> GTTGAAGCCATACACC</td><td> 3-10-3 MOE</td><td> 20</td>

<td> 372896</td><td> 432</td><td> 445</td><td> TTGAAGCCATACAC</td><td> 2-10-2 MOE</td><td> 21</td>

<td> 372819</td><td> 438</td><td> 453</td><td> CCTCAGGGTTGAAGCC</td><td> 3-10-3 MOE</td><td> 22</td>

<td> 372897</td><td> 439</td><td> 452</td><td> CTCAGGGTTGAAGC</td><td> 2-10-2 MOE</td><td> 23</td>

<td> 372820</td><td> 443</td><td> 458</td><td> TTTGCCCTCAGGGTTG</td><td> 3-10-3 MOE</td><td> 24</td>

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<td> 372898</td><td> 444</td><td> 457</td><td> TTGCCCTCAGGGTT</td><td> 2-10-2 MOE</td><td> 25</td>

<td> 372821</td><td> 468</td><td> 483</td><td> AGTTCTTGGTTTTCTT</td><td> 3-10-3 MOE</td><td> 26</td>

<td> 372899</td><td> 469</td><td> 482</td><td> GTTCTTGGTTTTCT</td><td> 2-10-2 MOE</td><td> 27</td>

<td> 372822</td><td> 587</td><td> 602</td><td> CCTCTTGATGTTCAGG</td><td> 3-10-3 MOE</td><td> 28</td>

<td> 372900</td><td> 588</td><td> 601</td><td> CTCTTGATGTTCAG</td><td> 2-10-2 MOE</td><td> 29</td>

<td> 372823</td><td> 592</td><td> 607</td><td> ATGCCCCTCTTGATGT</td><td> 3-10-3 MOE</td><td> 30</td>

<td> 372901</td><td> 593</td><td> 606</td><td> TGCCCCTCTTGATG</td><td> 2-10-2 MOE</td><td> 31</td>

<td> 346583</td><td> 715</td><td> 728</td><td> TGCCACATTGCCCT</td><td> 3-8-3 MOE</td><td> 32</td>

<td> 346584</td><td> ’716</td><td> 729</td><td> TTGCCACATTGCCC</td><td> 3-8-3 MOE</td><td> 33</td>

<td> 346585</td><td> 717</td><td> 730</td><td> GTTGCCACATTGCC</td><td> 3-8-3 MOE</td><td> 34</td>

<td> 346586</td><td> 718</td><td> 731</td><td> TGTTGCCACATTGC</td><td> 3-8-3 MOE</td><td> 35</td>

<td> 346587</td><td> 719</td><td> 732</td><td> CTGTTGCCACATTG</td><td> 3-8-3 MOE</td><td> 36</td>

<td> 346588</td><td> 720</td><td> 733</td><td> TCTGTTGCCACATT</td><td> 3-8-3 MOE</td><td> 37</td>

<td> 346589</td><td> 721</td><td> 734</td><td> TTCTGTTGCCACAT</td><td> 3-8-3 MOE</td><td> 38</td>

<td> 346590</td><td> 722</td><td> 735</td><td> TTTCTGITGCCACA</td><td> 3-8-3 MOE</td><td> 39</td>

<td> 346591</td><td> 723</td><td> 736</td><td> ATTTCTGTTGCCAC</td><td> 3-8-3 MOE</td><td> 40</td>

<td> 372824</td><td> 929</td><td> 944</td><td> GTAGGAGAAAGGCAGG</td><td> 3-10-3 MOE</td><td> 41</td>

<td> 372902</td><td> 930</td><td> 943</td><td> TAGGAGAAAGGCAG</td><td> 2-10-2 MOE</td><td> 42</td>

<td> 372825</td><td> 1256</td><td> 1271</td><td> GGCTTGTAAAGTGATG</td><td> 3-10-3 MOE</td><td> 43</td>

<td> 372903</td><td> 1257</td><td> 1270</td><td> GCTTGTAAAGTGAT</td><td> 2-10-2 MOE</td><td> 44</td>

<td> 372826</td><td> 1304</td><td> 1319</td><td> CCACTGGAGGATGTGA</td><td> 3-10-3 MOE</td><td> 45</td>

<td> 372904</td><td> 1305</td><td> 1318</td><td> CACTGGAGGATGTG</td><td> 2-10-2 MOE</td><td> 46</td>

<td> 372829</td><td> 2135</td><td> 2150</td><td> TTTCAGCATGCTTTCT</td><td> 3-10-3 MOE</td><td> 47</td>

<td> 372907</td><td> 2136</td><td> 2149</td><td> TTCAGCATGCTTTC</td><td> 2-10-2 MOE</td><td> 48</td>

<td> 372832</td><td> 2774</td><td> 2789</td><td> CATATTTGTCACAAAC</td><td> 3-10-3 MOE</td><td> 49</td>

<td> 372910</td><td> 2775</td><td> 2788</td><td> ATATTTGTCACAAA</td><td> 2-10-2 MOE</td><td> 50</td>

<td> 372833</td><td> 2779</td><td> 2794</td><td> ATGCCCATAITTGTCA</td><td> 3-10-3 MOE</td><td> 51</td>

<td> 372911</td><td> 2780</td><td> 2793</td><td> TGCCCATATTTGTC</td><td> 2-10-2 MOE</td><td> 52</td>

<td> 372835</td><td> 2961</td><td> 2976</td><td> TTTTGGTGGTAGAGAC</td><td> 3-10-3 MOE</td><td> 53</td>

<td> 372913</td><td> 2962</td><td> 2975</td><td> TTTGGTGGTAGAGA</td><td> 2-10-2 MOE</td><td> 54</td>

<td> 346592</td><td> 3248</td><td> 3261</td><td> TCTGCTTCGCACCT</td><td> 3-8-3 MOE</td><td> 55</td>

<td> 346593</td><td> 3249</td><td> 3262</td><td> GTCTGCTTCGCACC</td><td> 3-8-3 MOE</td><td> 56</td>

<td> 346594</td><td> 3250</td><td> 3263</td><td> AGTCTGCTTCGCAC</td><td> 3-8-3 MOE</td><td> 57</td>

<td> 346595</td><td> 3251</td><td> 3264</td><td> CAGTCTGCTTCGCA</td><td> 3-8-3 MOE</td><td> 58</td>

<td> 346596</td><td> 3252</td><td> 3265</td><td> TCAGTCTGCTTCGC</td><td> 3-8-3 MOE</td><td> 59</td>

<td> 346597</td><td> 3253</td><td> 3266</td><td> CTCAGTCTGCTTCG</td><td> 3-8-3 MOE</td><td> 60</td>

<td> 346598</td><td> 3254</td><td> 3267</td><td> CCTCAGTCTGCTTC</td><td> 3-8-3 MOE</td><td> 61</td>

<td> 346599</td><td> 3255</td><td> 3268</td><td> GCCTCAGTCTGCTT</td><td> 3-8-3 MOE</td><td> 62</td>

<td> 346600</td><td> 3256</td><td> 3269</td><td> AGCCTCAGTCTGCT</td><td> 3-8-3 MOE</td><td> 63</td>

<td> 372836</td><td> 3350</td><td> 3365</td><td> AACTCTGAGGATTGTT</td><td> 3-10-3 MOE</td><td> 64</td>

<td> 372914</td><td> 3351</td><td> 3364</td><td> ACTCTGAGGATTGT</td><td> 2-10-2 MOE</td><td> 65</td>

<td> 372837</td><td> 3355</td><td> 3370</td><td> TCATTAACTCTGAGGA</td><td> 3-10-3 MOE</td><td> 66</td>

<td> 372915</td><td> 3356</td><td> 3369</td><td> CATTAACTCTGAGG</td><td> 2-10-2 MOE</td><td> 67</td>

<td> 372838</td><td> 3360</td><td> 3375</td><td> ATTCATCATTAACTCT</td><td> 3-10-3 MOE</td><td> 68</td>

<td> 372916</td><td> 3361</td><td> 3374</td><td> TTCATCATTAACTC</td><td> 2-10-2 MOE</td><td> 69</td>

<td> 372839</td><td> 3409</td><td> 3424</td><td> TTGTTCTGAATGTCCA</td><td> 3-10-3 MOE</td><td> 70</td>

<td> 387461</td><td> 3409</td><td> 3424</td><td> TTGTTCTGAATGTCCA</td><td> 3-10-3 Methyleneoxy BNA Unmodified cytosines in gap</td><td> 70</td>

<td> 380147</td><td> 3409</td><td> 3424</td><td> TTGTTCTGAATGTCCA</td><td> 3-10-3 Methyleneoxy BNA</td><td> 70</td>

<td> 372917</td><td> 3410</td><td> 3423</td><td> TGTTCTGAATGTCC</td><td> 2-10-2 MOE</td><td> 73</td>

<td> 372840</td><td> 3573</td><td> 3588</td><td> CAGATGAGTCCATTTG</td><td> 3-10-3 MOE</td><td> 74</td>

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<td> 372918</td><td> 3574</td><td> 3587</td><td> AGATGAGTCCATTT</td><td> 2-10-2 MOE</td><td> 75</td>

<td> 372841</td><td> 3701</td><td> 3716</td><td> ATCCACAGGGAAATTG</td><td> 3-10-3 MOE</td><td> 76</td>

<td> 372919</td><td> 3702</td><td> 3715</td><td> TCCACAGGGAAATT</td><td> 2-10-2 MOE</td><td> 77</td>

<td> 372843</td><td> 4219</td><td> 4234</td><td> CAGTTGTACAAGTTGC</td><td> 3-10-3 MOE</td><td> 78</td>

<td> 372921</td><td> 4220</td><td> 4233</td><td> AGTTGTACAAGTTG</td><td> 2-10-2 MOE</td><td> 79</td>

<td> 372844</td><td> 4301</td><td> 4316</td><td> CACAGAGTCAGCCTTC</td><td> 3-10-3 MOE</td><td> 80</td>

<td> 372922</td><td> 4302</td><td> 4315</td><td> ACAGAGTCAGCCTT</td><td> 2-10-2 MOE</td><td> 81</td>

<td> 372845</td><td> 4308</td><td> 4323</td><td> GGTCAACCACAGAGTC</td><td> 3-10-3 MOE</td><td> 82</td>

<td> 372923</td><td> 4309</td><td> 4322</td><td> GTCAACCACAGAGT</td><td> 2-10-2 MOE</td><td> 83</td>

<td> 346601</td><td> 5588</td><td> 5601</td><td> CAGCCACATGCAGC</td><td> 3-8-3 MOE</td><td> 84</td>

<td> 346602</td><td> 5589</td><td> 5602</td><td> CCAGCCACATGCAG</td><td> 3-8-3 MOE</td><td> 85</td>

<td> 346603</td><td> 5590</td><td> 5603</td><td> ACCAGCCACATGCA</td><td> 3-8-3 MOE</td><td> 86</td>

<td> 346604</td><td> 5591</td><td> 5604</td><td> TACCAGCCACATGC</td><td> 3-8-3 MOE</td><td> 87</td>

<td> 346605</td><td> 5592</td><td> 5605</td><td> TTACCAGCCACATG</td><td> 3-8-3 MOE</td><td> 88</td>

<td> 346606</td><td> 5593</td><td> 5606</td><td> GTTACCAGCCACAT</td><td> 3-8-3 MOE</td><td> 89</td>

<td> 346607</td><td> 5594</td><td> 5607</td><td> GGTTACCAGCCACA</td><td> 3-8-3 MOE</td><td> 90</td>

<td> 346608</td><td> 5595</td><td> 5608</td><td> AGGTTACCAGCCAC</td><td> 3-8-3 MOE</td><td> 91</td>

<td> 346609</td><td> 5596</td><td> 5609</td><td> TAGGTTACCAGCCA</td><td> 3-8-3 MOE</td><td> 92</td>

<td> 372851</td><td> 5924</td><td> 5939</td><td> AGGTTCTGCTTTCAAC</td><td> 3-10-3 MOE</td><td> 93</td>

<td> 372929</td><td> 5925</td><td> 5938</td><td> GGITCTGCTTTCAA</td><td> 2-10-2 MOE</td><td> 94</td>

<td> 372854</td><td> 6664</td><td> 6679</td><td> TACTGATCAAATTGTA</td><td> 3-10-3 MOE</td><td> 95</td>

<td> 372932</td><td> 6665</td><td> 6678</td><td> ACTGATCAAATTGT</td><td> 2-10-2 MOE</td><td> 96</td>

<td> 372855</td><td> 6908</td><td> 6923</td><td> TTTTTCTTGTATCTGG</td><td> 3-10-3 MOE</td><td> 97</td>

<td> 372933</td><td> 6909</td><td> 6922</td><td> TTTTCTTGTATCTG</td><td> 2-10-2 MOE</td><td> 98</td>

<td> 372856</td><td> 7190</td><td> 7205</td><td> ATCCATTAAAACCTGG</td><td> 3-10-3 MOE</td><td> 99</td>

<td> 372934</td><td> 7191</td><td> 7204</td><td> TCCATTAAAACCTG</td><td> 2-10-2 MOE</td><td> 100</td>

<td> 372858</td><td> 7817</td><td> 7832</td><td> ATATTGCTCTGCAAAG</td><td> 3-10-3 MOE</td><td> 101</td>

<td> 372936</td><td> 7818</td><td> 7831</td><td> TATTGCTCTGCAAA</td><td> 2-10-2 MOE</td><td> 102</td>

<td> 346610</td><td> 7818</td><td> 7831</td><td> TATTGCTCTGCAAA</td><td> 3-8-3 MOE</td><td> 102</td>

<td> 346611</td><td> 7819</td><td> 7832</td><td> ATATTGCTCTGCAA</td><td> 3-8-3 MOE</td><td> 104</td>

<td> 346612</td><td> 7820</td><td> 7833</td><td> AATATTGCTCTGCA</td><td> 3-8-3 MOE</td><td> 105</td>

<td> 346613</td><td> 7821</td><td> 7834</td><td> GAATATTGCTCTGC</td><td> 3-8-3 MOE</td><td> 106</td>

<td> 346614</td><td> 7822</td><td> 7835</td><td> AGAATATTGCTCTG</td><td> 3-8-3 MOE</td><td> 107</td>

<td> 346615</td><td> 7823</td><td> 7836</td><td> TAGAATATTGCTCT</td><td> 3-8-3 MOE</td><td> 108</td>

<td> 346616</td><td> 7824</td><td> 7837</td><td> ATAGAATATTGCTC</td><td> 3-8-3 MOE</td><td> 109</td>

<td> 346617</td><td> 7825</td><td> 7838</td><td> GATAGAATATTGCT</td><td> 3-8-3 MOE</td><td> 110</td>

<td> 346618</td><td> 7826</td><td> 7839</td><td> GGATAGAATATTGC</td><td> 3-8-3 MOE</td><td> 111</td>

<td> 372859</td><td> 7995</td><td> 8010</td><td> ATGGAATCCTCAAATC</td><td> 3-10-3 MOE</td><td> 112</td>

<td> 372937</td><td> 7996</td><td> 8009</td><td> TGGAATCCTCAAAT</td><td> 2-10-2 MOE</td><td> 113</td>

<td> 372861</td><td> 8336</td><td> 8351</td><td> GAATTCTGGTATGTGA</td><td> 3-10-3 MOE</td><td> 114</td>

<td> 372939</td><td> 8337</td><td> 8350</td><td> AATTCTGGTATGTG</td><td> 2-10-2 MOE</td><td> 115</td>

<td> 372862</td><td> 8341</td><td> 8356</td><td> AGCTGGAATTCTGGTA</td><td> 3-10-3 MOE</td><td> 116</td>

<td> 372940</td><td> 8342</td><td> 8355</td><td> GCTGGAATTCTGGT</td><td> 2-10-2 MOE</td><td> 117</td>

<td> 372863</td><td> 8539</td><td> 8554</td><td> TGAAAATCAAAATTGA</td><td> 3-10-3 MOE</td><td> 118</td>

<td> 372941</td><td> 8540</td><td> 8553</td><td> GAAAATCAAAATTG</td><td> 2-10-2 MOE</td><td> 119</td>

<td> 372871</td><td> 9344</td><td> 9359</td><td> AAACAGTGCATAGTTA</td><td> 3-10-3 MOE</td><td> 120</td>

<td> 372949</td><td> 9345</td><td> 9358</td><td> AACAGTGCATAGTT</td><td> 2-10-2 MOE</td><td> 121</td>

<td> 372872</td><td> 9515</td><td> 9530</td><td> TTCAGGAATTGTTAAA</td><td> 3-10-3 MOE</td><td> 122</td>

<td> 372950</td><td> 9516</td><td> 9529</td><td> TCAGGAATTGTTAA</td><td> 2-10-2 MOE</td><td> 123</td>

<td> 372875</td><td> 9794</td><td> 9809</td><td> TTTTGTTTCATTATAG</td><td> 3-10-3 MOE</td><td> 124</td>

<td> 372953</td><td> 9795</td><td> 9808</td><td> TTTGTTTCATTATA</td><td> 2-10-2 MOE</td><td> 125</td>

<td> 372877</td><td> 10157</td><td> 10172</td><td> GATGACACTTGATTTA</td><td> 3-10-3 MOE</td><td> 126</td>

<td> 372955</td><td> 10158</td><td> 10171</td><td> ATGACACTTGATTT</td><td> 2-10-2 MOE</td><td> 127</td>

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<td> 372878</td><td> 10161</td><td> 10176</td><td> GTGTGATGACACTTGA</td><td> 3-10-3 MOE</td><td> 128</td>

<td> 372956</td><td> 10162</td><td> 10175</td><td> TGTGATGACACTTG</td><td> 2-10-2 MOE</td><td> 129</td>

<td> 372879</td><td> 10167</td><td> 10182</td><td> TATTCAGTGTGATGAC</td><td> 3-10-3 MOE</td><td> 130</td>

<td> 372957</td><td> 10168</td><td> 10181</td><td> ATTCAGTGTGATGA</td><td> 2-10-2 MOE</td><td> 131</td>

<td> 372880</td><td> 10172</td><td> 10187</td><td> ATTGGTATTCAGTGTG</td><td> 3-10-3 MOE</td><td> 132</td>

<td> 372958</td><td> 10173</td><td> 10186</td><td> TTGGTATTCAGTGT</td><td> 2-10-2 MOE</td><td> 133</td>

<td> 346619</td><td> 10838</td><td> 10851</td><td> CCTCTAGCTGTAAG</td><td> 3-8-3 MOE</td><td> 134</td>

<td> 346620</td><td> 10839</td><td> 10852</td><td> CCCTCTAGCTGTAA</td><td> 3-8-3 MOE</td><td> 135</td>

<td> 346621</td><td> 10840</td><td> 10853</td><td> GCCCTCTAGCTGTA</td><td> 3-8-3 MOE</td><td> 136</td>

<td> 346622</td><td> 10841</td><td> 10854</td><td> GGCCCTCTAGCTGT</td><td> 3-8-3 MOE</td><td> 137</td>

<td> 346623</td><td> 10842</td><td> 10855</td><td> AGGCCCTCTAGCTG</td><td> 3-8-3 MOE</td><td> 138</td>

<td> 346624</td><td> 10843</td><td> 10856</td><td> GAGGCCCTCTAGCT</td><td> 3-8-3 MOE</td><td> 139</td>

<td> 346625</td><td> 10844</td><td> 10857</td><td> AGAGGCCCTCTAGC</td><td> 3-8-3 MOE</td><td> 140</td>

<td> 346626</td><td> 10845</td><td> 10858</td><td> AAGAGGCCCTCTAG</td><td> 3-8-3 MOE</td><td> 141</td>

<td> 346627</td><td> 10846</td><td> 10859</td><td> AAAGAGGCCCTCTA</td><td> 3-8-3 MOE</td><td> 142</td>

<td> 372890</td><td> 13689</td><td> 13704</td><td> GAATGGACAGGTCAAT</td><td> 3-10-3 MOE</td><td> 143</td>

<td> 372968</td><td> 13690</td><td> 13703</td><td> AATGGACAGGTCAA</td><td> 2-10-2 MOE</td><td> 144</td>

<td> 372891</td><td> 13694</td><td> 13709</td><td> GTTTTGAATGGACAGG</td><td> 3-10-3 MOE</td><td> 145</td>

<td> 372969</td><td> 13695</td><td> 13708</td><td> TTTTGAATGGACAG</td><td> 2-10-2 MOE</td><td> 146</td>

<td> 372892</td><td> 13699</td><td> 13714</td><td> TGGTAGTTTTGAATGG</td><td> 3-10-3 MOE</td><td> 147</td>

<td> 372970</td><td> 13700</td><td> 13713</td><td> GGTAGTTTTGAATG</td><td> 2-10-2 MOE</td><td> 148</td>

<td> 346628</td><td> 13907</td><td> 13920</td><td> TCACTGTATGGTTT</td><td> 3-8-3 MOE</td><td> 149</td>

<td> 346629</td><td> 13908</td><td> 13921</td><td> CTCACTGTATGGTT</td><td> 3-8-3 MOE</td><td> 150</td>

<td> 346630</td><td> 13909</td><td> 13922</td><td> GCTCACTGTATGGT</td><td> 3-8-3 MOE</td><td> 151</td>

<td> 346631</td><td> 13910</td><td> 13923</td><td> GGCTCACTGTATGG</td><td> 3-8-3 MOE</td><td> 152</td>

<td> 346632</td><td> 13911</td><td> 13924</td><td> TGGCTCACTGTATG</td><td> 3-8-3 MOE</td><td> 153</td>

<td> 346633</td><td> 13912</td><td> 13925</td><td> CTGGCTCACTGTAT</td><td> 3-8-3 MOE</td><td> 154</td>

<td> 346634</td><td> 13913</td><td> 13926</td><td> GCTGGCTCACTGTA</td><td> 3-8-3 MOE</td><td> 155</td>

<td> 346635</td><td> 13914</td><td> 13927</td><td> GGCTGGCTCACTGT</td><td> 3-8-3 MOE</td><td> 156</td>

<td> 346636</td><td> 13915</td><td> 13928</td><td> AGGCTGGCTCACTG</td><td> 3-8-3 MOE</td><td> 157</td>

<td> 346637</td><td> 13963</td><td> 13976</td><td> CAGGTCCAGTTCAT</td><td> 3-8-3 MOE</td><td> 158</td>

<td> 346638</td><td> 13964</td><td> 13977</td><td> GCAGGTCCAGTTCA</td><td> 3-8-3 MOE</td><td> 159</td>

<td> 346639</td><td> 13965</td><td> 13978</td><td> TGCAGGTCCAGTTC</td><td> 3-8-3 MOE</td><td> 160</td>

<td> 346640</td><td> 13966</td><td> 13979</td><td> GTGCAGGTCCAGTT</td><td> 3-8-3 MOE</td><td> 161</td>

<td> 346641</td><td> 13967</td><td> 13980</td><td> GGTGCAGGTCCAGT</td><td> 3-8-3 MOE</td><td> 162</td>

<td> 346642</td><td> 13968</td><td> 13981</td><td> TGGTGCAGGTCCAG</td><td> 3-8-3 MOE</td><td> 163</td>

<td> 346643</td><td> 13969</td><td> 13982</td><td> TTGGTGCAGGTCCA</td><td> 3-8-3 MOE</td><td> 164</td>

<td> 346644</td><td> 13970</td><td> 13983</td><td> TTTGGTGCAGGTCC</td><td> 3-8-3 MOE</td><td> 165</td>

<td> 346645</td><td> 13971</td><td> 13984</td><td> CTTTGGTGCAGGTC</td><td> 3-8-3 MOE</td><td> 166</td>

<td> 346646</td><td> 14051</td><td> 14064</td><td> TAACTCAGATCCTG</td><td> 3-8-3 MOE</td><td> 167</td>

<td> 346647</td><td> 14052</td><td> 14065</td><td> ATAACTCAGATCCT</td><td> 3-8-3 MOE</td><td> 168</td>

<td> 346648</td><td> 14053</td><td> 14066</td><td> AATAACTCAGATCC</td><td> 3-8-3 MOE</td><td> 169</td>

<td> 346649</td><td> 14054</td><td> 14067</td><td> AAATAACTCAGATC</td><td> 3-8-3 MOE</td><td> 170</td>

<td> 346650</td><td> 14055</td><td> 14068</td><td> AAAATAACTCAGAT</td><td> 3-8-3 MOE</td><td> 171</td>

<td> 346651</td><td> 14056</td><td> 14069</td><td> CAAAATAACTCAGA</td><td> 3-8-3 MOE</td><td> 172</td>

<td> 346652</td><td> 14057</td><td> 14070</td><td> GCAAAATAACTCAG</td><td> 3-8-3 MOE</td><td> 173</td>

<td> 346653</td><td> 14058</td><td> 14071</td><td> AGCAAAATAACTCA</td><td> 3-8-3 MOE</td><td> 174</td>

<td> 346654</td><td> 14059</td><td> 14072</td><td> TAGCAAAATAACTC</td><td> 3-8-3 MOE</td><td> 175</td>

Table 3: Short antisense compounds targeted to SEQ ID NO: 1 and having 1 or 2 mismatches

<td></td><td> 5’</td><td> 3’</td><td></td><td></td><td> SEQ</td>

<td> Isis</td><td> Target</td><td> Target</td><td></td><td></td><td> ID</td>

<td> NO.</td><td> Site</td><td> Site</td><td> Sequence (5’-3’)</td><td> Gapmer Motif</td><td> NO</td>

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<td> 372894</td><td> 771</td><td> 784</td><td> CGGAGGTGCTTGAA</td><td> 2-10-2 MOE</td><td> 17</td>

<td> 372905</td><td> 1111</td><td> 1124</td><td> CAGGGCCTGGAGAG</td><td> 2-10-2 MOE</td><td> 176</td>

<td> 346628</td><td> 1493</td><td> 1506</td><td> TCACTGTATGGTTT</td><td> 3-8-3 MOE</td><td> 149</td>

<td> 372828</td><td> 2006</td><td> 2021</td><td> TCTGAAGTCCATGATC</td><td> 3-10-3 MOE</td><td> 177</td>

<td> 372906</td><td> 2007</td><td> 2020</td><td> CTGAAGTCCATGAT</td><td> 2-10-2 MOE</td><td> 178</td>

<td> 372830</td><td> 2382</td><td> 2397</td><td> TGGGCATGATTCCATT</td><td> 3-10-3 MOE</td><td> 179</td>

<td> 372908</td><td> 2383</td><td> 2396</td><td> GGGCATGATTCCAT</td><td> 2-10-2 MOE</td><td> 180</td>

<td> 346616</td><td> 3162</td><td> 3175</td><td> ATAGAATATTGCTC</td><td> 3-8-3 MOE</td><td> 109</td>

<td> 346617</td><td> 3163</td><td> 3176</td><td> GATAGAATATTGCT</td><td> 3-8-3 MOE</td><td> 110</td>

<td> 372929</td><td> 3513</td><td> 3526</td><td> GGTTCTGCTTTCAA</td><td> 2-10-2 MOE</td><td> 94</td>

<td> 372946</td><td> 3800</td><td> 3813</td><td> TGGAGCCCACGTGC</td><td> 2-10-2 MOE</td><td> 181</td>

<td> 372904</td><td> 4040</td><td> 4053</td><td> CACTGGAGGATGTG</td><td> 2-10-2 MOE</td><td> 46</td>

<td> 372842</td><td> 4084</td><td> 4099</td><td> TTGAAGTTGAGGGCTG</td><td> 3-10-3 MOE</td><td> 182</td>

<td> 372920</td><td> 4085</td><td> 4098</td><td> TGAAGTTGAGGGCT</td><td> 2-10-2 MOE</td><td> 183</td>

<td> 346586</td><td> 4778</td><td> 4791</td><td> TGTTGCCACATTGC</td><td> 3-8-3 MOE</td><td> 35</td>

<td> 372847</td><td> 5030</td><td> 5045</td><td> ACCAGTATTAATTTTG</td><td> 3-10-3 MOE</td><td> 184</td>

<td> 372925</td><td> 5031</td><td> 5044</td><td> CCAGTATTAATTTT</td><td> 2-10-2 MOE</td><td> 185</td>

<td> 372848</td><td> 5192</td><td> 5207</td><td> GTGTTCTTTGAAGCGG</td><td> 3-10-3 MOE</td><td> 186</td>

<td> 372926</td><td> 5193</td><td> 5206</td><td> TGTTCTTTGAAGCG</td><td> 2-10-2 MOE</td><td> 187</td>

<td> 372953</td><td> 5625</td><td> 5638</td><td> TTTGTTTCATTATA</td><td> 2-10-2 MOE</td><td> 125</td>

<td> 372935</td><td> 7585</td><td> 7598</td><td> AGTTACTTTGGTGT</td><td> 2-10-2 MOE</td><td> 188</td>

<td> 372860</td><td> 8255</td><td> 8270</td><td> TGGTACATGGAAGTCT</td><td> 3-10-3 MOE</td><td> 189</td>

<td> 372938</td><td> 8256</td><td> 8269</td><td> GGTACATGGAAGTC</td><td> 2-10-2 MOE</td><td> 190</td>

<td> 391260</td><td> 8256</td><td> 8269</td><td> GGTACATGGAAGTC</td><td> 2-10-2 MOE</td><td> 190</td>

<td> 392068</td><td> 8256</td><td> 8269</td><td> GGTACATGGAAGTC</td><td> 2-10-2 MOE</td><td> 190</td>

<td> 387462</td><td> 8256</td><td> 8269</td><td> GGTACATGGAAGTC</td><td> 2-10-2 Methyleneoxy BNA</td><td> 190</td>

<td> 391872</td><td> 8256</td><td> 8269</td><td> GGTACATGGAAGTC</td><td> 1-1-10-22’(butylacetomido)palmitamide Methyleneoxy BNA/Methyleneoxy BNA Unmodified cytosines in gap</td><td> 190</td>

<td> 380148</td><td> 8256</td><td> 8269</td><td> GGTACATGGAAGTC</td><td> 2-10-2 Methyleneoxy BNA</td><td> 190</td>

<td> 391871</td><td> 8256</td><td> 8269</td><td> GGTACATGGAAGTC</td><td> 1-1-10-2 2’(butylacetomido)palmitamide/MOE/MOE Unmodified cytosines in gap</td><td> 190</td>

<td> 391755</td><td> 8256</td><td> 8269</td><td> GGTACATGGAAGTC</td><td> 2-10-2 ENA mC in wing only</td><td> 190</td>

<td> 398296</td><td> 8256</td><td> 8269</td><td> GGTACATGGAAGTC</td><td> 2-10-2 (6’S)-6’-methylMethyleneoxy BNA Unmodified Cytosines</td><td> 190</td>

<td> 372942</td><td> 8455</td><td> 8468</td><td> TCCATGCCATATGT</td><td> 2-10-2 MOE</td><td> 200</td>

<td> 372865</td><td> 8888</td><td> 8903</td><td> CCCTGAAGAAGTCCAT</td><td> 3-10-3 MOE</td><td> 201</td>

<td> 372943</td><td> 8889</td><td> 8902</td><td> CCTGAAGAAGTCCA</td><td> 2-10-2 MOE</td><td> 202</td>

<td> 372866</td><td> 8908</td><td> 8923</td><td> GCCCAGTTCCATGACC</td><td> 3-10-3 MOE</td><td> 203</td>

<td> 372944</td><td> 8909</td><td> 8922</td><td> CCCAGTTCCATGAC</td><td> 2-10-2 MOE</td><td> 204</td>

<td> 372867</td><td> 9058</td><td> 9073</td><td> TTGAGGAAGCCAGATT</td><td> 3-10-3 MOE</td><td> 205</td>

<td> 372945</td><td> 9059</td><td> 9072</td><td> TGAGGAAGCCAGAT</td><td> 2-10-2 MOE</td><td> 206</td>

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<td> 372870</td><td> 9261</td><td> 9276</td><td> TGGATGCAGTAATCTC</td><td> 3-10-3 MOE</td><td> 207</td>

<td> 372948</td><td> 9262</td><td> 9275</td><td> GGATGCAGTAATCT</td><td> 2-10-2 MOE</td><td> 208</td>

<td> 372881</td><td> 10185</td><td> 10200</td><td> TATAAAGTCCAGCATT</td><td> 3-10-3 MOE</td><td> 209</td>

<td> 372959</td><td> 10186</td><td> 10199</td><td> ATAAAGTCCAGCAT</td><td> 2-10-2 MOE</td><td> 210</td>

<td> 372882</td><td> 10445</td><td> 10460</td><td> AAGTTCCTGCTTGAAG</td><td> 3-10-3 MOE</td><td> 211</td>

<td> 372960</td><td> 10446</td><td> 10459</td><td> AGTTCCTGCTTGAA</td><td> 2-10-2 MOE</td><td> 212</td>

<td> 372964</td><td> 11451</td><td> 11464</td><td> AATGGTGAAGTACT</td><td> 2-10-2 MOE</td><td> 213</td>

<td> 346612</td><td> 13459</td><td> 13472</td><td> AATATTGCTCTGCA</td><td> 3-8-3 MOE</td><td> 105</td>

<td> 346613</td><td> 13460</td><td> 13473</td><td> GAATATTGCTCTGC</td><td> I^- 3-8-3 MOE</td><td> 106</td>

In certain embodiments, a target region is nucleotides 263-278 of SEQ ID NO: 1. In certain such embodiments, short antisense compounds targeted to nucleotides 263-278 of SEQ ID NO: 1 comprise a nucleotide sequence selected from SEQ ID NO: 16 or 17. In certain such embodiments, a short antisense compound targeted to nucleotides 263-278 of SEQ ID NO: 1 is selected from Isis NO. 372816 or 372894.

In certain embodiments, a target region is nucleotides 428-483 of SEQ ID NO: 1. In certain such embodiments, a short antisense compound targeted to nucleotides 428-483 of SEQ ID NO: 1 comprises a nucleotide sequence selected from SEQ ID NO 18, 19,20,21, 22, 23,24, 25, 26, or 27. In certain such embodiments, a short antisense compound targeted to nucleotides 428-483 of SEQ ID NO: 1 is selected from Isis NO. 372817,372895,372818,372896,372819,372897,372820,372898,372821, or 372899.

In certain embodiments, a target region is nucleotides 428-458 of SEQ ID NO: 1. In certain such embodiments, a short antisense compound targeted to nucleotides 428-458 of SEQ ID NO: I comprises a nucleotide sequence selected from SEQ ID NO 18, 19, 20, 21, 22, 23, 24, or 25. In certain such embodiments, a short antisense compound targeted to nucleotides 428-458 of SEQ ID NO: 1 is selected from Isis NO. 372817,372895,372818,372896,372819,372897,372820, or 372898.

In certain embodiments, a target region is nucleotides 468-483 of SEQ ID NO: 1. In certain such embodiments, a short antisense compound targeted to nucleotides 468-483 of SEQ ID NO: 1 comprises a nucleotide sequence selected from SEQ ID NO 26 or 27. In certain such embodiments, a short antisense compound targeted to nucleotides 468-483 of SEQ ID NO: 1 is selected from Isis NO. 372821 or 372899.

In certain embodiments, a target region is nucleotides 587-607 of SEQ ID NO: 1. In certain such embodiments, a short antisense compound targeted to nucleotides 587-607 of SEQ ID NO: 1 comprises a nucleotide sequence selected from SEQ ID NO 28, 29, 30, or 31. In certain such embodiments, a short antisense compound targeted to nucleotides 587-607 of SEQ ID NO: 1 is selected from ISIS NO. 372822, 372900,372823, or 372901.

In certain embodiments, a target region is nucleotides 715-736 of SEQ ID NO: 1. In certain such embodiments, a short antisense compound targeted to nucleotides 715-736 of SEQ ID NO: 1 comprises a nucleotide sequence selected from SEQ ID NO 32, 33, 34, 35, 36, 37, 38, 39, or 40. In certain such embodiments, a short antisense compound targeted to nucleotides 715-736 of SEQ ID NO: 1 is selected from Isis NO. 346583,346584,346585,346586, 346587,346588,346589,346590, or 346591.

In certain embodiments, a target region is nucleotides 929-944 of SEQ ID NO: 1. In certain such embodiments, a short antisense compound targeted to nucleotides 929-944 of SEQ ID NO: 1 comprises a

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-59nucleotide sequence selected from SEQ ID NO 41 or 42. In certain such embodiments, a short antisense compound targeted to nucleotides 929-944 of SEQ ID NO: 1 is selected from Isis NO. 372824 or 372902.

In certain embodiments, a target region is nucleotides 1256-1319 of SEQ ID NO: 1. In certain such embodiments, a short antisense compound targeted to nucleotides 1256-1319 of SEQ ID NO: I comprises a nucleotide sequence selected from SEQ ID NO 43, 44, 45, or 46. In certain such embodiments, a short antisense compound targeted to nucleotides 1256-1319 of SEQ ID NO: 1 is selected from Isis NO. 372825,372903,372826, or 372904.

In certain embodiments, a target region is nucleotides 1256-1271 of SEQ ID NO: 1. In certain such embodiments, a short antisense compound targeted to nucleotides 1256-1271 of SEQ ID NO: 1 comprises a nucleotide sequence selected from SEQ ID NO 43 or 44. In certain such embodiments, a short antisense compound targeted to nucleotides 1256-1271 of SEQ ID NO: 1 is selected from Isis NO. 372825 or 372903.

In certain embodiments, a target region is nucleotides 1304-1319 of SEQ ID NO; 1. In certain such embodiments, a short antisense compound targeted to nucleotides 1304-1319 of SEQ ID NO: 1 comprises a nucleotide sequence selected from SEQ ID NO 45 or 46. In certain such embodiments, a short antisense compound targeted to nucleotides 1304-1319 of SEQ ID NO: 1 is selected from Isis NO. 372826 or 372904.

In certain embodiments, a target region is nucleotides 2135-2150 of SEQ ID NO: 1. In certain such embodiments, a short antisense compound targeted to nucleotides 2135-2150 of SEQ ID NO: 1 comprises a nucleotide sequence selected from SEQ ID NO 47 or 48. In certain such embodiments, a short antisense compound targeted to nucleotides 2135-2150 of SEQ ID NO: I is selected from ISIS NO. 372829 or 372907.

In certain embodiments, a target region is nucleotides 2774-2794 of SEQ ID NO: 1. In certain such embodiments, a short antisense compound targeted to nucleotides 2774-2794 of SEQ ID NO: 1 comprises a nucleotide sequence selected from SEQ ID NO 49, 50, 51, or 52. In certain such embodiments, a short antisense compound targeted to nucleotides 2774-2794 of SEQ ID NO: 1 is selected from ISIS NO. 372832,372910,372833, or 372911.

In certain embodiments, a target region is nucleotides 2961-2976 of SEQ ID NO: 1. In certain such embodiments, a short antisense compound targeted to nucleotides 2961-2976 of SEQ ID NO: I comprises a nucleotide sequence selected from SEQ ID NO 53 or 54. In certain such embodiments, a short antisense compound targeted to nucleotides 2961-2976 of SEQ ID NO: 1 is selected from ISIS NO. 372835 or 372913.

In certain embodiments, a target region is nucleotides 3248-3269 of SEQ ID NO: 1. In certain such embodiments, a short antisense compound targeted to nucleotides 3248-3269 of SEQ ID NO: 1 comprises a nucleotide sequence selected from SEQ ID NO 55,56,57,58,59,60,61,62, or 63. In certain such embodiments, a short antisense compound targeted to nucleotides 3248-3269 of SEQ ID NO: 1 is selected from ISIS NO. 346592,346593,346594,346595,346596,346597,346598, 346599, or 346600.

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-60In certain embodiments, a target region is nucleotides 3350-3375 of SEQ ID NO: 1. In certain such embodiments, a short antisense compound targeted to nucleotides 33S0-3375 of SEQ ID NO: 1 comprises a nucleotide sequence selected from SEQ ID NO 64, 65, 66, 67, 68, or 69. In certain such embodiments, a short antisense compound targeted to nucleotides 3350-3375 of SEQ ID NO: 1 is selected from ISIS NO. 372836,372914,372837,372915,372838, or 372916.

In certain embodiments, a target region is nucleotides 3409-3424 of SEQ ID NO: 1. In certain such embodiments, a short antisense compound targeted to nucleotides 3409-3424 of SEQ ID NO: 1 comprises a nucleotide sequence selected from SEQ ID NO 70 or 73. In certain such embodiments, a short antisense compound targeted to nucleotides 3409-3424 of SEQ ID NO: 1 is selected from ISIS NO. 372839,387461,380147, or 372917.

In certain embodiments, a target region is nucleotides 3573-3588 of SEQ ID NO: 1. In certain such embodiments, a short antisense compound targeted to nucleotides 3573-3588 of SEQ ID NO: 1 comprises a nucleotide sequence selected from SEQ ID NO 74 or 75. In certain such embodiments, a short antisense compound targeted to nucleotides 3573-3588 of SEQ ID NO: 1 is selected from ISIS NO. 372840 or 372918.

In certain embodiments, a target region is nucleotides 3701-3716 of SEQ ID NO: 1. In certain such embodiments, a short antisense compound targeted to nucleotides 3701-3716 of SEQ ID NO: 1 comprises a nucleotide sequence selected from SEQ ID NO 76 or 77. In certain such embodiments, a short antisense compound targeted to nucleotides 3701-3716 of SEQ ID NO: 1 is selected from ISIS NO. 372841 or 372919.

In certain embodiments, a target region is nucleotides 4219-4234 of SEQ ID NO: 1. In certain such embodiments, a short antisense compound targeted to nucleotides 4219-4234 of SEQ ID NO: 1 comprises a nucleotide sequence selected from SEQ ID NO 78 or 79. In certain such embodiments, a short antisense compound targeted to nucleotides 4219-4234 of SEQ ID NO: 1 is selected from ISIS NO. 372843 or 372921.

In certain embodiments, a target region is nucleotides 4301-4323 of SEQ ID NO: 1. In certain such embodiments, a short antisense compound targeted to nucleotides 4301-4323 of SEQ ID NO: 1 comprises a nucleotide sequence selected from SEQ ID NO 80, 81, 82, or 83. In certain embodiments, a short antisense compound targeted to nucleotides 4301-4323 of SEQ ID NO: 1 is selected from ISIS NO. 372844,372922,372845, or 372923.

In certain embodiments, a target region is nucleotides 5588-5609 of SEQ ID NO: 1. In certain such embodiments, a short antisense compound targeted to nucleotides 5588-5609 of SEQ ID NO: 1 comprises a nucleotide sequence selected from SEQ ID NO 84,85,86,87,88,89,90,91, or 92. In certain such embodiments, a short antisense compound targeted to nucleotides 5588-5609 of SEQ ID NO: 1 is selected from ISIS NO. 346601,346602,346603,346604,346605,346606,346607,346608, or 346609.

In certain embodiments, a target region is nucleotides 5924-5939 of SEQ ID NO: 1. In certain such embodiments, a short antisense compound targeted to nucleotides 5924-5939 of SEQ ID NO; 1

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-61comprises a nucleotide sequence selected from SEQ ID NO 93 or 94. In certain such embodiments, a short antisense compound targeted to nucleotides 5924-5939 of SEQ ID NO: 1 is selected from ISIS NO. 372851 or 372929.

In certain embodiments, a target region is nucleotides 6664-6679 of SEQ ID NO: 1. In certain such embodiments, a short antisense compound targeted to nucleotides 6664-6679 of SEQ ID NO: 1 comprises a nucleotide sequence selected from SEQ ID NO 95 or 96. In certain such embodiments, a short antisense compound targeted to nucleotides 6664-6679 of SEQ ID NO: 1 is selected from ISIS NO. 372854 or 372932.

In certain embodiments, a target region is nucleotides 6908-6923 of SEQ ID NO: 1. In certain such embodiments, a short antisense compound targeted to nucleotides 6908-6923 of SEQ ID NO: I comprises a nucleotide sequence selected from SEQ ID NO 97 or 98. In certain such embodiments, a short antisense compound targeted to nucleotides 6908-6923 of SEQ ID NO: 1 is selected from ISIS NO. 372855 or 372933.

In certain embodiments, a target region is nucleotides 7190-7205 of SEQ ID NO: 1. In certain such embodiments, a short antisense compound targeted to nucleotides 7190-7205 of SEQ ID NO: 1 comprises a nucleotide sequence selected from SEQ ID NO 99 or 100. In certain such embodiments, a short antisense compound targeted to nucleotides 7190-7205 of SEQ ID NO: 1 is selected from ISIS NO. 372856 or 372934.

In certain embodiments, a target region is nucleotides 7817-7839 of SEQ ID NO: 1. In certain such embodiments, a short antisense compound targeted to nucleotides 7817-7839 of SEQ ID NO: 1 comprises a nucleotide sequence selected from SEQ ID NO 101,102,104,105, 106,107,108,109, 110, or 111. In certain such embodiments, a short antisense compound targeted to nucleotides 7817-7839 of SEQ ID NO: 1 is selected from ISIS NO. 372858, 372936, 346610, 346611, 346612, 346613, 346614, 346615,346616,346617, or 346618.

In certain embodiments, a target region is nucleotides 7995-8010 of SEQ ID NO: 1. In certain such embodiments, a short antisense compound targeted to nucleotides 7995-8010 of SEQ ID NO: 1 comprises a nucleotide sequence selected from SEQ ID NO 112 or 113. In certain such embodiments, a short antisense compound targeted to nucleotides 7995-8010 of SEQ ID NO: I is selected from ISIS NO. 372859 or 372937.

In certain embodiments, a target region is nucleotides 8336-8356 of SEQ ID NO: 1. In certain such embodiments, a short antisense compound targeted to nucleotides 8336-8356 of SEQ ID NO: 1 comprises a nucleotide sequence selected from SEQ ID NO 114, 115, 116, or 117. In certain such embodiments, a short antisense compound targeted to nucleotides 8336-8356 of SEQ ID NO: 1 is selected from ISIS NO. 372861,372939,372862, or 372940.

In certain embodiments, a target region is nucleotides 8539-8554 of SEQ ID NO: 1. In certain such embodiments, a short antisense compound targeted to nucleotides 8539-8554 of SEQ ID NO: 1 comprises a nucleotide sequence selected from SEQ ID NO 118 or 119. In certain such embodiments, a

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-62short antisense compound targeted to nucleotides 8539-8554 of SEQ ID NO: 1 is selected from ISIS NO. 372863 or 372941.

In certain embodiments, a target region is nucleotides 9344-9359 of SEQ ID NO: 1. In certain such embodiments, a short antisense compound targeted to nucleotides 9344-9359 of SEQ ID NO: 1 comprises a nucleotide sequence selected from SEQ ID NO 120 or 121. In certain such embodiments, a short antisense compound targeted to nucleotides 9344-9359 of SEQ ID NO: 1 is selected from ISIS NO. 372871 or 372949.

In certain embodiments, a target region is nucleotides 9515-9530 of SEQ ID NO: 1. In certain such embodiments, a short antisense compound targeted to nucleotides 9515-9530 of SEQ ID NO: 1 comprises a nucleotide sequence selected from SEQ ID NO 122 or 123. In certain such embodiments, a short antisense compound targeted to nucleotides 9515-9530 of SEQ ID NO: 1 is selected from ISIS NO. 372872 or 372950.

In certain embodiments, a target region is nucleotides 9794-9809 of SEQ ID NO: 1. In certain such embodiments, a short antisense compound targeted to nucleotides 9794-9809 of SEQ ID NO: 1 comprises a nucleotide sequence selected from SEQ ID NO 124 or 125. In certain such embodiments, a short antisense compound targeted to nucleotides 9794-9809 of SEQ ID NO: 1 is selected from ISIS NO. 372875 or 372953.

In certain embodiments, a target region is nucleotides 10157-10187 ofSEQ ID NO: 1. Incertain such embodiments, a short antisense compound targeted to nucleotides 10157-10187 of SEQ ID NO: 1 comprises a nucleotide sequence selected from SEQ ID NO 126,127,128,129, 130,131,132, or 133. In certain such embodiments, a short antisense compound targeted to nucleotides 10157-10187 of SEQ ID NO: 1 is selected from ISIS NO. 372877,372955,372878,372956,372879,372957,372880, or 372958.

In certain embodiments, a target region is nucleotides 10838-10859 of SEQ ID NO: 1. In certain such embodiments, a short antisense compound targeted to nucleotides 10838-10859 of SEQ ID NO: 1 comprises a nucleotide sequence selected from SEQ ID NO 134, 135, 136, 137, 138, 139, 140, 141, or 142. In certain such embodiments, a short antisense compound targeted to nucleotides 10838-10859 of SEQ ID NO; 1 is selected from ISIS NO. 346619, 346620, 346621, 346622, 346623, 346624, 346625, 346626, or 346627.

In certain embodiments, a target region is nucleotides 13689-13714 of SEQ ID NO: 1. In certain such embodiments, a short antisense compound targeted to nucleotides 13689-13714 of SEQ ID NO: 1 comprises a nucleotide sequence selected from SEQ ID NO 143, 144, 145, 146, 147, or 148. In certain such embodiments, a short antisense compound targeted to nucleotides 13689-13714 ofSEQ ID NO: 1 is selected from ISIS NO. 372890,372968,372891,372969,372892, or 372970.

In certain embodiments, a target region is nucleotides 13907-13928 of SEQ ID NO: 1. In certain such embodiments, a short antisense compound targeted to nucleotides 13907-13928 of SEQ ID NO: 1 comprises a nucleotide sequence selected from SEQ ID NO 149, 150, 151,152, 153, 154, 155, 156, or 157. In certain such embodiments, a short antisense compound targeted to nucleotides 13907-13928 of

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-63SEQ ID NO: 1 is selected from ISIS NO. 346628, 346629, 346630, 346631, 346632, 346633, 346634, 346635, or 346636.

In certain embodiments, a target region is nucleotides 13963-13984 of SEQ ID NO: 1. In certain such embodiments, a short antisense compound targeted to nucleotides 13963-13984 of SEQ ID NO: 1 comprises a nucleotide sequence selected from SEQ ID NO 158, 159, 160, 161, 162, 163, 164, 165, or 166. In certain such embodiments, a short antisense compound targeted to nucleotides 13963-13984 of SEQ ID NO: 1 is selected from ISIS NO. 346637, 346638, 346639, 346640, 346641, 346642, 346643, 346644, or 346645.

In certain embodiments, a target region is nucleotides 14051-14072 of SEQ ID NO: 1. In certain such embodiments, a short antisense compound targeted to nucleotides 14051-14072 of SEQ ID NO: 1 comprises a nucleotide sequence selected from SEQ ID NO 167, 168, 169, 170, 171, 172, 173, 174, or 175. In certain such embodiments, a short antisense compound targeted to nucleotides 14051-14072 of SEQ ID NO: 1 is selected from ISIS NO. 346646, 346647, 346648, 346649, 346650, 346651, 346652, 346653, or 346654.

In certain embodiments, short antisense compounds targeted to an ApoB nucleic acid are 8 to 16, preferably 9 to 15, more preferably 9 to 14, more preferably 10 to 14 nucleotides in length. In certain embodiments, short antisense compounds targeted to an ApoB nucleic acid are 9 to 14 nucleotides in length. In certain embodiments, short antisense compounds targeted to an ApoB nucleic acid are 10 to 14 nucleotides in length. In certain embodiments, such short antisense compounds are short antisense oligonucleotides.

In certain embodiments, short antisense compounds targeted to an ApoB nucleic acid are short gapmers. In certain such embodiments, short gapmers targeted to an ApoB nucleic acid comprise at least one high affinity modification in one or more wings of the compound. In certain embodiments, short antisense compounds targeted to an ApoB nucleic acid comprise 1 to 3 high-affinity modifications in each wing. In certain such embodiments, the nucleosides or nucleotides of the wing comprise a 2’ modification. In certain such embodiments, the monomers of the wing are BNA’s. In certain such embodiments, the monomers of the wing are selected from α-L-Methyleneoxy (4’-CH₂-O-2*) BNA, β-DMethyleneoxy (4’-CH₂-O-2’) BNA , Ethyleneoxy (4’-(CH₂)₂-O-2’) BNA , Aminooxy (4’-CH₂-O-N(R)2’) BNA and Oxyamino (4’-CHrN(R)-O-2’) BNA. In certain embodiments, the monomers of a wing comprise a substituent at the 2’ position selected from allyl, amino, azido, thio, O-allyl, 0-C|-Cio alkyl, OCFj, O-fCH^-O-CHj, 2'-O(CH₂)₂SCH₃,0-(CH₂)₂-0-N(R_ra)(R„), and O-CH₂-C(=O)-N(R_m)(R_n), where each R,, and R, is, independently, H or substituted or unsubstituted CpCio alkyl. In certain embodiments, the monomers of a wing are 2’MOE nucleotides.

In certain embodiments, short antisense compounds targeted to an ApoB nucleic acid comprise a gap between the 5’ wing and the 3’ wing. In certain embodiments the gap comprises five, six, seven, eight, nine, ten, eleven, twelve, thirteen, or fourteen monomers. In certain embodiments, the monomers of the gap are unmodified deoxyribonucleotides. In certain embodiments, the monomers of the gap are

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-64unmodified ribonucleotides. In certain embodiments, gap modifications (if any) gap result in an antisense compound that, when bound to its target nucleic acid, supports cleavage by an RNase, including, but not limited to, RNase H.

In certain embodiments, short antisense compounds targeted to an ApoB nucleic acid have uniform monomeric linkages. In certain such embodiments, those linkages are all phosphorothioate linkages. In certain embodiments, the linkages are all phosphodiester linkages. In certain embodiments, short antisense compounds targeted to an ApoB nucleic acid have mixed backbones.

In certain embodiments, short antisense compounds targeted to an ApoB nucleic acid are 8 monomers in length. In certain embodiments, short antisense compounds targeted to an ApoB nucleic acid are 9 monomers in length. In certain embodiments, short antisense compounds targeted to an ApoB nucleic acid are 10 monomers in length. In certain embodiments, short antisense compounds targeted to an ApoB nucleic acid are 11 monomers in length. In certain embodiments, short antisense compounds targeted to an ApoB nucleic acid are monomers in length. In certain embodiments, short antisense compounds targeted to an ApoB nucleic acid are 13 monomers in length. In certain embodiments, short antisense compounds targeted to an ApoB nucleic acid are 14 monomers in length. In certain embodiments, short antisense compounds targeted to an ApoB nucleic acid are 15 monomers in length. In certain embodiments, short antisense compounds targeted to an ApoB nucleic acid are 16 monomers in length. In certain embodiments, short antisense compounds targeted to an ApoB nucleic acid comprise 9 to 15 monomers. In certain embodiments, short antisense compounds targeted to an ApoB nucleic acid comprise 10 to 15 monomers. In certain embodiments, short antisense compounds targeted to an ApoB nucleic acid comprise 12 to 14 monomers. In certain embodiments, short antisense compounds targeted to an ApoB nucleic acid comprise 12 to 14 nucleotides or nucleosides.

In certain embodiments, the invention provides methods of modulating expression of ApoB. In certain embodiments, such methods comprise use of one or more short antisense compound targeted to an ApoB nucleic acid, wherein the short antisense compound targeted to an ApoB nucleic acid is from about 8 to about 16, preferably 9 to 15, more preferably 9 to 14, more preferably 10 to 14 monomers (i.e. from about 8 to about 16 linked monomers). One of ordinary skill in the art will appreciate that this comprehends methods of modulating expression of ApoB using one or more short antisense compounds targeted to an ApoB nucleic acid of 8,9,10,11,12,13,14,15 or 16 monomers.

In certain embodiments, methods of modulating ApoB comprise use of a short antisense compound targeted to an ApoB nucleic acid that is 8 monomers in length. In certain embodiments, methods of modulating ApoB comprise use of a short antisense compound targeted to an ApoB nucleic acid that is 9 monomers in length. In certain embodiments, methods of modulating ApoB comprise use of a short antisense compound targeted to an ApoB nucleic acid that is 10 monomers in length. In certain embodiments, methods of modulating ApoB comprise use of a short antisense compound targeted to an ApoB nucleic acid that is 11 monomers in length. In certain embodiments, methods of modulating ApoB comprise use of a short antisense compound targeted to an ApoB nucleic acid that is 12 monomers in

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-65length. In certain embodiments, methods of modulating ApoB comprise use of a short antisense compound targeted to an ApoB nucleic acid that is 13 monomers in length. In certain embodiments, methods of modulating ApoB comprise use of a short antisense compound targeted to an ApoB nucleic acid that is 14 monomers in length. In certain embodiments, methods of modulating ApoB comprise use of a short antisense compound targeted to an ApoB nucleic acid that is 15 monomers in length. In certain embodiments, methods of modulating ApoB comprise use of a short antisense compound targeted to an ApoB nucleic acid that is 16 monomers in length.

In certain embodiments, methods of modulating expression of ApoB comprise use of a short antisense compound targeted to an ApoB nucleic acid comprising 9 to 15 monomers. In certain embodiments, methods of modulating expression of ApoB comprise use of a short antisense compound targeted to an ApoB nucleic acid comprising 10 to 15 monomers. In certain embodiments, methods of modulating expression of ApoB comprise use of a short antisense compound targeted to an ApoB nucleic acid comprising 12 to 14 monomers. In certain embodiments, methods of modulating expression of ApoB comprise use of a short antisense compound targeted to an ApoB nucleic acid comprising 12 or 14 nucleotides or nucleosides.

In certain embodiments, short antisense compounds targeting a ApoB nucleic acid may have any one or more properties or characteristics of the short antisense compounds generally described herein. In certain embodiments, short antisense compounds targeting a ApoB nucleic acid have a motif (wing deoxy gap -wing) selected from 1-12-1,1-1-10-2,2-10-1-1,3-10-3,2-10-3,2-10-2,1-10-1,1-10-2, 3-83,2-8-2,1-8-1,3-6-3 or 1-6-1, more preferably 1-10-1,2-10-2,3-10-3, and 1-9-2.

2. SGLT-2

Sodium dependent glucose transporter 2 (SGLT-2) is expressed in the kidney proximal tubule epithelial cells, and functions to reabsorb glucose preventing glucose loss in the urine. For the human genome SGLT-2 is a member of an 11-membered family of sodium substrate co-transporters. Many of these family members share sequence homology, for example SGLT-1 shares about 59% sequence identity with SGLT-2 and about 70% sequence identity with SGLT-3. SGLT-1 is a glucose transporter found in the heart and the CNS. SGLT-3 is a glucose sensing sodium channel in the small intestine. The separate localization patterns for these SGLTs is one point of distinction between the homologous family members. (Handion, AL., Expert Opin. Ther. Patents (2005) 15(ll):1532-1540; Kanai et al., J. Clin. Invest., 1994,93,397-404; Wells ct al., Am. J. Physiol. Endocrinol. Metab., 1992,263, F459-465).

Studies of human SGLT2 injected into Xenopus oocytes demonstrated that this protein mediates sodium-dependent transport of D-glucose and .alpha.-methyl-D-glucopyranoside (,alpha.-MeGlc; a glucose analog) with a Km value of 1.6 mM for .alpha.-MeGlc and a sodium to glucose coupling ratio of 1:1 (Kanai et al., J. Clin. Invest., 1994,93,397-404; You et al., J. Biol. Chem., 1995,270,29365-29371). This transport activity was suppressed by phlorizin, a plant glycoside that binds to the glucose site of the

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-66SGLTs but is not transported and thus inhibits SGLT action (You et al., J. Biol. Chem., 1995,270,2936529371).

Diabetes is a disorder characterized by hyperglycemia due to deficient insulin action. Chronic hyperglycemia is a major risk factor for diabetes-associated complications, including heart disease, retinopathy, nephropathy and neuropathy. As the kidneys play a major role in the regulation of plasma glucose levels, renal glucose transporters are becoming attractive drug targets (Wright, Am. J. Physiol. Renal Physiol., 2001,280, F10-18). Diabetic nephropathy is the most common cause of end-stage renal disease that develops in many patients with diabetes. Glucotoxicity, which results from long-term hyperglycemia, induces tissue-dependent insulin resistance in diabetic patients (Nawano et al., Am. J. Physiol. Endocrinol. Metab., 2000,278, E535-543).

Definitions “Sodium dependent glucose transporter 2” is the gene product or protein of which expression is to be modulated by administration of a short antisense compound. Sodium dependent glucose transporter 2 is generally referred to as SGLT2 but may also be referred to as SLC5A2; sodium-glucose transporter 2; sodium-glucose cotransporter, kidney low affinity; sodium-glucose cotransporter, renal; solute carrier family 5 (sodium/glucose cotransporter), member 2; SL52.

“SGLT2 nucleic acid means any nucleic acid encoding SGLT2. For example, in certain embodiments, a SGLT2 nucleic acid includes, without limitation, a DNA sequence encoding SGLT2, an RNA sequence transcribed from DNA encoding SGLT2, and an mRNA sequence encoding SGLT2. “SGLT2 mRNA” means an mRNA encoding a SGLT2 protein.

Therapeutic indications

In certain embodiments, short antisense compounds are used to modulate expression of SGLT-2 and related proteins. In certain embodiments, such modulation is accomplished by providing short antisense compounds that hybridize with one or more target nucleic acid molecules encoding SGLT-2, including, but is not limited to, SGLT2, SL52, SLC5A2, Sodium-Glucose Co-Transporter, Kidney Low Affinity Sodium-Glucose Co-Transporter, Renal Sodium-Glucose Co-Transporter 2 and Solute Carrier Family 5 Sodium/Glucose Co-Transporter Member 2. Also provided are methods of treating metabolic and/or cardiovascular disease and disorders as described herein. In particular embodiments, short antisense compounds that inhibit the expression of SGLT2 are used in methods of lowering blood glucose levels in an animal and methods of delaying or preventing the onset of type 2 diabetes. Such methods comprise administering a therapeutically or prophylactically effective amount of one or more of the compounds of the invention to the animal, which may be in need of treatment. The one or more compounds can be a short antisense compound targeting a nucleic acid encoding SGLT2. Provided herein are methods of enhancing inhibition of expression of SGLT2 in kidney cells or kidney tissues, comprising contacting the cells or tissues with one or more of the compounds of the invention, such as short antisense compounds targeting a nucleic

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-67acid encoding SGLT2.

While certain compounds, compositions and methods have been described with specificity in accordance with certain embodiments, the following examples serve only to illustrate the compounds of the invention and are not intended to limit the same.

In certain embodiments, short antisense compounds are chimeric oligomeric compounds having mixed phosphorothioate and phosphodiester backbones,. Certain mixed backbone short antisense compounds have a central gap comprising at least 5 contiguous 2'-deoxy nucleosides flanked by two wings each of which comprises at least one 2'-O-methoxyethyl nucleoside. In certain embodiments, the intemucleoside linkages of the mixed backbone compounds are phosphorothioate linkages in the gap and phosphodiester linkages in the two wings. In certain embodiments, mixed backbone compounds have phosphorothioate linkages in the wings, except for one phosphodiester linkage at one or both of the extreme 5' and 3' ends of the oligonucleotide. In certain embodiments short antisense compounds targeted to SGLT2 have a motif (wing - deoxy gap -wing) selected from 3-10-3, 2-10-3, 2-10-2, 1-101,1-10-2, 2-8-2,1-9-2,1-8-1,3-6-3 or 1-6-1. In certain embodiments short antisense compounds targeted to SGLT2 have a motif (wing-deoxy gap -wing) selected from 1-10-1,1-10-2,2-8-2,1-9-2,1-8-1,3-6-3 or 1-6-1.

In certain embodiments, short antisense compounds targeted to an SGLT2 nucleic acid and having a mixed backbone are efficiently delivered to the kidney. In certain embodiments, administration of short antisense compounds targeted to an SGLT2 nucleic acid and having a mixed backbone results in modulation of target gene expression in the kidney. In certain such embodiments, there is little or no liver or kidney toxicity. In certain embodiments, short antisense compounds targeted to an SGLT2 nucleic acid and having a mixed backbone are more potent for reducing SGLT-2 mRNA and have a faster onset compared with a short antisense compound that does not have a mixed back-bone, but is otherwise identical. In certain such embodiments, such increase potency and/or reduced toxicity is in mouse and/or rat. In certain such embodiments, such increase potency and/or reduced toxicity is in a human.

By way of example, and only for illustrative purposes, ISIS 145733, which comprises uniform phosphorothioate linkages and ISIS 257016 which comprises phosphodiester linkage in the wings and phosphorothioate linkages in the gap, are otherwise identical. Both comprise the sequence GAAGTAGCCACCAACTGTGC (SEQ ID NO. 1572). Both of the oligonucleotides further comprise a gap consisting of ten 2'-deoxynucleotides, flanked on each side by five-nucleotide “2’-methoxyethyl (2MOE) nucleotides. All cytidine residues are 5-methylcytidines. The mixed back-bone compound, ISIS 257016, was about 50 times more potent for reducing SGLT-2 mRNA compared to the non-mixed parent compound, ISIS 145733 (see EXAMPLE 9).

Pharmacokinetic studies of certain mixed backbone compound ISIS 257016 indicate that in certain embodiments, the compound acts as a prodrug that is metabolized to a 12 nucleobase pharmacophore. Studies with ISIS 370717, a 12 nucleobase short antisense compound corresponding to

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-68ISIS 257016, show that the compound has a similar pharmacological profile to ISIS 257016 but with a faster onset of action. ISIS 370717 is a 12 nucleobase antisense oligonucleotide targeted to SGLT-2 comprising the sequence TAGCCACCAACT (SEQ ID NO. 1554), further comprising a gap consisting of ten 2'-deoxynucleotide$, flanked on both sides by one-nucleotide wings. The wings are composed of 2'-methoxyethyl (2-MOE) nucleotides. All cytidine residues are 5-methylcytidines. The intemucleoside linkages are phosphorothioate (P=S) throughout the oligonucleotide. The similarity in pharmacological activity of ISIS 257016 and ISIS 370717 supports the pharmacokinetic studies indicating ISIS 257016 was a prodrug having a 12 nucleotide pharmacophore (see EXAMPLE 10). Further, studies with stabilized (end-capped) versions of ISIS 257016 show dramatic loss of activity.

In certain embodiments, short antisense compounds comprising 2’ MOE monomers in the wings are efficiently delivered to the kidney and treatment with such compounds results in efficient modulation of target gene expression in the kidney without liver or kidney toxicity. It is further shown herein that in certain embodiments, short antisense compounds are more potent for reducing SGLT-2 mRNA and have a faster onset compared with parent oligonucleotides targeted to SGLT-2 mRNA in mouse and rat. 2’ MOE gap shortmers are shown herein to improve potency and bioavailability over parent compounds.

By way of example, and only for illustrative purposes studies with ISIS 370717 reveal significantly higher accumulation of the short antisense compound in the kidney tissue (approximately 500 micro grams per gram of tissue) compared to the longer parent. Moreover, SGLT-2 mRNA was reduced by more than 80% over the controls (see EXAMPLE 11). ISIS 370717 1-10-1 gapmer was used as a template to make sequence related oligos with varying motifs. Studies evaluating wing, gap and total length variations around the ISIS 370717 12 mer oligonucleotide can be seen in EXAMPLE 12. Certain motifs evaluated included 1-10-1, 2-8-2, 1-8-1, 3-6-3, and 1-6-1 (see Table 60 in EXAMPLE 12). The compounds were analyzed for their effect on SGLT2 mRNA levels. All the motifs inhibited the expression of SGLT2 in vivo in a dose-dependent manner. The 1-10-1, 2-8-2 and 1-8-1 gapmers were found to be particularly potent. SGLT-2 mRNA was reduced by more than 80% over the controls using these motifs.

In certain embodiments, the invention provides short antisense compounds targeted to an SGLT2 nucleic acid and having a motif selected from: 1-10-1 and 1-10-2 MOE gapmer. (see Table 62 in EXAMPLE 13). Certain such compounds were analyzed for their effect on rat SGLT2 mRNA. Results in Table 63 illustrate that both the 1-10-1 and 1-10-2 MOE gapmers inhibit the expression of SGLT2 in vivo in a dose-dependent manner and over 80% reduction of SGLT-2 mRNA could be achieved.

Certain additional 1-10-1 and 2-8-2 MOE gapmers were evaluated in both mouse and rat in vivo models (see, e g., EXAMPLE 14 and 15). Greater than 80% reduction in SGLT-2 mRNA was achieved with many of the 1-10-1 and 2-8-2 MOE gapmers at relatively low concentrations of oligo and in the absence of any toxicity effects.

In another non-limiting example, the effect of ISIS 388625 on dog SGLT2 mRNA levels was also analyzed. Dog studies illustrate that greater than 80% inhibition of the expression of SGLT2 can be

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-69achieved at a I mg/kg/wk dose. Even greater inhibition can be achieved at slightly higher doses. Administration of ISIS 388625 in dog was also shown to improved glucose tolerance. Peak plasma glucose levels were decreased by over 50% on average and the subsequent drop in glucose was lessened compared to saline controls in a standard glucose tolerance test (See EXAMPLE 17). Also, in a rat model of diabetes, short antisense compounds were shown to significantly decrease plasma glucose levels and HbAlC over time compared to PBS and control treated animals (See Example 16).

The animals in all studies were further evaluated for toxicity. For example, total body weight, liver, spleen and kidney weight were evaluated. Significant changes in spleen, liver or body weight can indicate that a particular compound causes toxic effects. All changes were found to be within the margin of error. No significant changes in body weight were observed during the treatment or at study termination. No significant changes in liver or spleen weights were observed.

Certain Short Antisense Compounds Targeted to an SGLT2 nucleic acid

In certain embodiments, short antisense compounds are targeted to an SGLT2 nucleic acid having the sequence of GENBANK® Accession No. NM 003041.1, incoiporated herein as SEQ ID NO: 2. In certain such embodiments, a short antisense compound targeted to SEQ ID NO: 3 is at least 90% complementary to SEQ ID NO: 3. In certain such embodiments, a short antisense compound targeted to SEQ ID NO: 3 is at least 95% complementary to SEQ ID NO: 3. In certain such embodiments, a short antisense compound targeted to SEQ ID NO: 3 is 100% complementary to SEQ ID NO: 1. In certain embodiments, a sheet antisense compound targeted to SEQ ID NO: 3 comprises a nucleotide sequence selected from the nucleotide sequences set forth in Table 4 and 5.

The nucleotide sequence set forth in each SEQ ID NO set forth in Tables 4 and 5 is independent of any modification to a sugar moiety, a monomeric linkage, or a nucleobase. As such, short antisense compounds defined by a SEQ ID NO may comprise, independently, one or more modifications to a sugar moiety, an intemucleoside linkage, or a nucleobase. Antisense compounds described by Isis Number (Isis NO.) indicate a combination of nucleobase sequence and one or more modifications to a sugar moiety, an intemucleoside linkage, or a nucleobase.

Tables 4 and 5 illustrate examples of short antisense compounds targeted to SEQ ID NO: 3. Table 4 illustrates short antisense compounds that are 100% complementary to SEQ ID NO: 3. Table 5 illustrates short antisense compounds that have one or two mismatches with respect to SEQ ID NO: 3. The column labeled ‘gapmer motif indicates the wing-gap-wing motif of each short antisense compounds. The gap segment comprises 2’-deoxynucleotides and each nucleotide of each wing segment comprises a 2’-modified sugar. The particular 2’-modified sugar is also indicated in the ‘gapmer motif column. For example, ‘2-10-2 MOE’ means a 2-10-2 gapmer motif, where a gap segment of ten 2’deoxynucleotides is flanked by wing segments of two nucleotides, where the nucleotides of the wing segments are 2’-MOE nucleotides. Intemucleoside linkages are phosphorothioate. The short antisense compounds comprise 5-methylcytidine in place of unmodified cytosine, unless “unmodified cytosine” is

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-70listed in the gapmer motif column, in which case the indicated cytosines are unmodified cytosines. For example, “5-mC in gap only” indicates that the gap segment has 5-methylcytosines, while the wing segments have unmodified cytosines.

Table 4: Short Antisense Compounds Targeted to SEQ ID NO: 3

<td> ISIS No</td><td> 5’ Target Site</td><td> 3’ Target Site</td><td> Sequence (S’-3’)</td><td> Gapmer Motif</td><td> SEQID NO</td>

<td> 379684</td><td> 84</td><td> 95</td><td> TGTCAGCAGGAT</td><td> 1-10-1 MOE</td><td> 214</td>

<td> 405193</td><td> 113</td><td> 124</td><td> CAGCAGGAAATA</td><td> 2-8-2 MOE</td><td> 215</td>

<td> 405194</td><td> 114</td><td> 125</td><td> CCAGCAGGAAAT</td><td> 2-8-2 MOE</td><td> 216</td>

<td> 405195</td><td> 115</td><td> 126</td><td> ACCAGCAGGAAA</td><td> 2-8-2 MOE</td><td> 217</td>

<td> 405196</td><td> 116</td><td> 127</td><td> GACCAGCAGGAA</td><td> 2-8-2 MOE</td><td> 218</td>

<td> 405197</td><td> 117</td><td> 128</td><td> TGACCAGCAGGA</td><td> 2-8-2 MOE</td><td> 219</td>

<td> 379685</td><td> 117</td><td> 128</td><td> TGACCAGCAGGA</td><td> 1-10-1 MOE</td><td> 219</td>

<td> 405198</td><td> 118</td><td> 129</td><td> ATGACCAGCAGG</td><td> 2-8-2 MOE</td><td> 221</td>

<td> 405199</td><td> 119</td><td> 130</td><td> AATGACCAGCAG</td><td> 2-8-2 MOE</td><td> 222</td>

<td> 405200</td><td> 120</td><td> 131</td><td> CAATGACCAGCA</td><td> 2-8-2 MOE</td><td> 223</td>

<td> 405201</td><td> 121</td><td> 132</td><td> CCAATGACCAGC</td><td> 2-8-2 MOE</td><td> 224</td>

<td> 379686</td><td> 135</td><td> 146</td><td> ACCACAAGCCAA</td><td> 1-10-1 MOE</td><td> 225</td>

<td> 379711</td><td> 172</td><td> 183</td><td> TAGCCGCCCACA</td><td> 1-10-1 MOE</td><td> 226</td>

<td> 388628</td><td> 172</td><td> 183</td><td> TAGCCGCCCACA</td><td> 2-8-2 MOE</td><td> 226</td>

<td> 405202</td><td> 207</td><td> 218</td><td> CCGGCCACCACA</td><td> 2-8-2 MOE</td><td> 228</td>

<td> 405203</td><td> 208</td><td> 219</td><td> ACCGGCCACCAC</td><td> 2-8-2 MOE</td><td> 229</td>

<td> 405204</td><td> 236</td><td> 247</td><td> GATGTTGCTGGC</td><td> 2-8-2 MOE</td><td> 230</td>

<td> 379687</td><td> 236</td><td> 247</td><td> GATGTTGCTGGC</td><td> 1-10-1 MOE</td><td> 230</td>

<td> 405205</td><td> 237</td><td> 248</td><td> CGATGTTGCTGG</td><td> 2-8-2 MOE</td><td> 232</td>

<td> 405206</td><td> 238</td><td> 249</td><td> CCGATGTTGCTG</td><td> 2-8-2 MOE</td><td> 233</td>

<td> 405207</td><td> 239</td><td> 250</td><td> GCCGATGTTGCT</td><td> 2-8-2 MOE</td><td> 234</td>

<td> 405208</td><td> 240</td><td> 251</td><td> TGCCGATGTTGC</td><td> 2-8-2 MOE</td><td> 235</td>

<td> 405209</td><td> 241</td><td> 252</td><td> CTGCCGATGTTG</td><td> 2-8-2 MOE</td><td> 236</td>

<td> 405210</td><td> 260</td><td> 271</td><td> CAGGCCCACAAA</td><td> 2-8-2 MOE</td><td> 237</td>

<td> 405211</td><td> 261</td><td> 272</td><td> CCAGGCCCACAA</td><td> 2-8-2 MOE</td><td> 238</td>

<td> 405212</td><td> 262</td><td> 273</td><td> GCCAGGCCCACA</td><td> 2-8-2 MOE</td><td> 239</td>

<td> 379688</td><td> 288</td><td> 299</td><td> CCAAGCCACTTG</td><td> 1-10-1 MOE</td><td> 240</td>

<td> 379689</td><td> 318</td><td> 329</td><td> AGAGCGCATTCC</td><td> 1-10-1 MOE</td><td> 241</td>

<td> 379690</td><td> 435</td><td> 446</td><td> ACAGGTAGAGGC</td><td> 1-10-1 MOE</td><td> 242</td>

<td> 405248</td><td> 474</td><td> 485</td><td> AGATCTTGGTGA</td><td> 2-8-2 MOE</td><td> 243</td>

<td> 379691</td><td> 474</td><td> 485</td><td> AGATCTTGGTGA</td><td> 1-10-1 MOE</td><td> 243</td>

<td> 382676</td><td> 527</td><td> 539</td><td> TGTTCCAGCCCAG</td><td> 1-10-2 MOE</td><td> 245</td>

<td> 388625</td><td> 528</td><td> 539</td><td> TGTTCCAGCCCA</td><td> 2-8-2 MOE</td><td> 246</td>

<td> 389780</td><td> 528</td><td> 539</td><td> TGTTCCAGCCCA</td><td> 1-9-2 MOE</td><td> 246</td>

<td> 379692</td><td> 528</td><td> 539</td><td> TGTTCCAGCCCA</td><td> 1-10-1 MOE</td><td> 246</td>

<td> 392170</td><td> 528</td><td> 539</td><td> TGTTCCAGCCCA</td><td> 1-10-1 Methyleneoxy BNA</td><td> 246</td>

<td> 392173</td><td> 528</td><td> 539</td><td> TGTTCCAGCCCA</td><td> 2-8-2 Methyleneoxy BNA</td><td> 246</td>

<td> 405213</td><td> 529</td><td> 540</td><td> ATGTTCCAGCCC</td><td> 2-8-2 MOE</td><td> 251</td>

<td> 405214</td><td> 564</td><td> 575</td><td> TGGTGATGCCCA</td><td> 2-8-2 MOE</td><td> 252</td>

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<td> 405215</td><td> 565</td><td> 576</td><td> ATGGTGATGCCC</td><td> 2-8-2 MOE</td><td> 253</td>

<td> 405216</td><td> 566</td><td> 577</td><td> CATGGTGATGCC</td><td> 2-8-2 MOE</td><td> 254</td>

<td> 379693</td><td> 566</td><td> 577</td><td> CATGGTGATGCC</td><td> 1-10-1 MOE</td><td> 254</td>

<td> 405217</td><td> 567</td><td> 578</td><td> TCATGGTGATGC</td><td> 2-8-2 MOE</td><td> 256</td>

<td> 405218</td><td> 568</td><td> 579</td><td> ATCATGGTGATG</td><td> 2-8-2 MOE</td><td> 257</td>

<td> 405219</td><td> 587</td><td> 598</td><td> CCCTCCTGTCAC</td><td> 2-8-2 MOE</td><td> 258</td>

<td> 405220</td><td> 588</td><td> 599</td><td> GCCCTCCTGTCA</td><td> 2-8-2 MOE</td><td> 259</td>

<td> 405221</td><td> 589</td><td> 600</td><td> AGCCCTCCTGTC</td><td> 2-8-2 MOE</td><td> 260</td>

<td> 405222</td><td> 590</td><td> 601</td><td> CAGCCCTCCTGT</td><td> 2-8-2 MOE</td><td> 261</td>

<td> 405223</td><td> 591</td><td> 602</td><td> CCAGCCCTCCTG</td><td> 2-8-2 MOE</td><td> 262</td>

<td> 405224</td><td> 592</td><td> 603</td><td> GCCAGCCCTCCT</td><td> 2-8-2 MOE</td><td> 263</td>

<td> 379694</td><td> 629</td><td> 640</td><td> GACGAAGGTCTG</td><td> 1-10-1 MOE</td><td> 264</td>

<td> 405225</td><td> 707</td><td> 718</td><td> GTATTTGTCGAA</td><td> 2-8-2 MOE</td><td> 265</td>

<td> 379695</td><td> 737</td><td> 748</td><td> GGACACCGTCAG</td><td> 1-10-1 MOE</td><td> 266</td>

<td> 379696</td><td> 974</td><td> 985</td><td> CAGCTTCAGGTA</td><td> 1-10-1 MOE</td><td> 267</td>

<td> 405226</td><td> 998</td><td> 1009</td><td> CATGACCATGAG</td><td> 2-8-2 MOE</td><td> 268</td>

<td> 405227</td><td> 999</td><td> 1010</td><td> GCATGACCATGA</td><td> 2-8-2 MOE</td><td> 269</td>

<td> 405228</td><td> 1000</td><td> 1011</td><td> GGCATGACCATG</td><td> 2-8-2 MOE</td><td> 270</td>

<td> 405229</td><td> 1001</td><td> 1012</td><td> TGGCATGACCAT</td><td> 2-8-2 MOE</td><td> 271</td>

<td> 405230</td><td> 1002</td><td> 1013</td><td> CTGGCATGACCA</td><td> 2-8-2 MOE</td><td> 272</td>

<td> 379697</td><td> 1002</td><td> 1013</td><td> CTGGCATGACCA</td><td> 1-10-1 MOE</td><td> 272</td>

<td> 405231</td><td> 1003</td><td> 1014</td><td> CCTGGCATGACC</td><td> 2-8-2 MOE</td><td> 274</td>

<td> 379698</td><td> 1091</td><td> 1102</td><td> GCAGCCCACCTC</td><td> 1-10-1 MOE</td><td> 275</td>

<td> 405232</td><td> 1092</td><td> 1103</td><td> AGCAGCCCACCT</td><td> 2-8-2 MOE</td><td> 276</td>

<td> 405233</td><td> 1093</td><td> 1104</td><td> GAGCAGCCCACC</td><td> 2-8-2 MOE</td><td> 277</td>

<td> 405234</td><td> 1130</td><td> 1141</td><td> CATGAGCTTCAC</td><td> 2-8-2 MOE</td><td> 278</td>

<td> 405235</td><td> 1131</td><td> 1142</td><td> GCATGAGCTTCA</td><td> 2-8-2 MOE</td><td> 279</td>

<td> 382677</td><td> 1131</td><td> 1143</td><td> GGCATGAGCTTCA</td><td> 1-10-2 MOE</td><td> 280</td>

<td> 388626</td><td> 1132</td><td> 1143</td><td> GGCATGAGCTTC</td><td> 2-8-2 MOE</td><td> 281</td>

<td> 379699</td><td> 1132</td><td> 1143</td><td> GGCATGAGCTTC</td><td> 1-10-1 MOE</td><td> 281</td>

<td> 405236</td><td> 1133</td><td> 1144</td><td> GGGCATGAGCTT</td><td> 2-8-2 MOE</td><td> 283</td>

<td> 405237</td><td> 1157</td><td> 1168</td><td> CAGCATGAGTCC</td><td> 2-8-2 MOE</td><td> 284</td>

<td> 405238</td><td> 1158</td><td> 1169</td><td> CCAGCATGAGTC</td><td> 2-8-2 MOE</td><td> 285</td>

<td> 379700</td><td> 1158</td><td> 1169</td><td> CCAGCATGAGTC</td><td> 1-10-1 MOE</td><td> 285</td>

<td> 405239</td><td> 1159</td><td> 1170</td><td> GCCAGCATGAGT</td><td> 2-8-2 MOE</td><td> 287</td>

<td> 379701</td><td> 1230</td><td> 1241</td><td> CCATGGTGAAGA</td><td> 1-10-1 MOE</td><td> 288</td>

<td> 405240</td><td> 1542</td><td> 1553</td><td> CACAGCTGCCCG</td><td> 2-8-2 MOE</td><td> 289</td>

<td> 405241</td><td> 1543</td><td> 1554</td><td> ACACAGCTGCCC</td><td> 2-8-2 MOE</td><td> 290</td>

<td> 405242</td><td> 1544</td><td> 1555</td><td> CACACAGCTGCC</td><td> 2-8-2 MOE</td><td> 291</td>

<td> 382678</td><td> 1544</td><td> 1556</td><td> GCACACAGCTGCC</td><td> 1-10-2 MOE</td><td> 292</td>

<td> 388627</td><td> 1545</td><td> 1556</td><td> GCACACAGCTGC</td><td> 2-8-2 MOE</td><td> 293</td>

<td> 379702</td><td> 1545</td><td> 1556</td><td> GCACACAGCTGC</td><td> 1-10-1 MOE</td><td> 293</td>

<td> 379703</td><td> 1701</td><td> 1712</td><td> GCCGGAGACTGA</td><td> 1-10-1 MOE</td><td> 295</td>

<td> 405243</td><td> 1976</td><td> 1987</td><td> ATTGAGGTTGAC</td><td> 2-8-2 MOE</td><td> 296</td>

<td> 405244</td><td> 1977</td><td> 1988</td><td> CATTGAGGTTGA</td><td> 2-8-2 MOE</td><td> 297</td>

<td> 405245</td><td> 1978</td><td> 1989</td><td> GCATTGAGGTTG</td><td> 2-8-2 MOE</td><td> 298</td>

<td> 405246</td><td> 1979</td><td> 1990</td><td> GGCATTGAGGTT</td><td> 2-8-2 MOE</td><td> 299</td>

<td> 405247</td><td> 1980</td><td> 1991</td><td> GGGCATTGAGGT</td><td> 2-8-2 MOE</td><td> 300</td>

Table 5: Short antisense compounds targeted to SEQ Π) NO: 3 and having 1 or 2 mismatches

<td></td><td> 5’</td><td> 3’</td><td></td><td></td><td></td>

<td> ISIS</td><td> Target</td><td> Target</td><td></td><td></td><td> SEQ ID</td>

<td> No</td><td> Site</td><td> Site</td><td> Sequence (5’-3’)</td><td> Gapmer Motif</td><td> NO</td>

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<td> 405200</td><td> 96</td><td> 107</td><td> CAATGACCAGCA</td><td> 2-8-2 MOE</td><td> 223</td>

<td> 405215</td><td> 382</td><td> 393</td><td> ATGGTGATGCCC</td><td> 2-8-2 MOE</td><td> 253</td>

<td> 405216</td><td> 383</td><td> 394</td><td> CATGGTGATGCC</td><td> 2-8-2 MOE</td><td> 254</td>

<td> 379693</td><td> 383</td><td> 394</td><td> CATGGTGATGCC</td><td> 1-10-1 MOE</td><td> 254</td>

<td> 379701</td><td> 471</td><td> 482</td><td> CCATGGTGAAGA</td><td> 1-10-1 MOE</td><td> 288</td>

<td> 405218</td><td> 472</td><td> 483</td><td> ATCATGGTGATG</td><td> 2-8-2 MOE</td><td> 257</td>

<td> 405246</td><td> 536</td><td> 547</td><td> GGCATTGAGGTT</td><td> 2-8-2 MOE</td><td> 299</td>

<td> 405248</td><td> 570</td><td> 581</td><td> AGATCTTGGTGA</td><td> 2-8-2 MOE</td><td> 243</td>

<td> 379691</td><td> 570</td><td> 581</td><td> AGATCTTGGTGA</td><td> 1-10-1 MOE</td><td> 243</td>

<td> 379698</td><td> 683</td><td> 694</td><td> GCAGCCCACCTC</td><td> 1-10-1 MOE</td><td> 275</td>

<td> 405232</td><td> 684</td><td> 695</td><td> AGCAGCCCACCT</td><td> 2-8-2 MOE</td><td> 276</td>

<td> 379711</td><td> 685</td><td> 696</td><td> TAGCCGCCCACA</td><td> 1-10-1 MOE</td><td> 226</td>

<td> 388628</td><td> 685</td><td> 696</td><td> TAGCCGCCCACA</td><td> 2-8-2 MOE</td><td> 226</td>

<td> 379698</td><td> 950</td><td> 961</td><td> GCAGCCCACCTC</td><td> 1-10-1 MOE</td><td> 275</td>

<td> 405232</td><td> 951</td><td> 962</td><td> AGCAGCCCACCT</td><td> 2-8-2 MOE</td><td> 276</td>

<td> 405235</td><td> 978</td><td> 989</td><td> GCATGAGCTTCA</td><td> 2-8-2 MOE</td><td> 279</td>

<td> 382677</td><td> 978</td><td> 990</td><td> GGCATGAGCTTCA</td><td> 1-10-2 MOE</td><td> 280</td>

<td> 388626</td><td> 979</td><td> 990</td><td> GGCATGAGCTTC</td><td> 2-8-2 MOE</td><td> 281</td>

<td> 379699</td><td> 979</td><td> 990</td><td> GGCATGAGCTTC</td><td> 1-10-1 MOE</td><td> 281</td>

<td> 405236</td><td> 980</td><td> 991</td><td> GGGCATGAGCTT</td><td> 2-8-2 MOE</td><td> 283</td>

<td> 379698</td><td> 1043</td><td> 1054</td><td> GCAGCCCACCTC</td><td> 1-10-1 MOE</td><td> 275</td>

<td> 405239</td><td> 1171</td><td> 1182</td><td> GCCAGCATGAGT</td><td> 2-8-2 MOE</td><td> 287</td>

<td> 405209</td><td> 1213</td><td> 1224</td><td> CTGCCGATGTTG</td><td> 2-8-2 MOE</td><td> 236</td>

<td> 405233</td><td> 1364</td><td> 1375</td><td> GAGCAGCCCACC</td><td> 2-8-2 MOE</td><td> 277</td>

<td> 405240</td><td> 1366</td><td> 1377</td><td> CACAGCTGCCCG</td><td> 2-8-2 MOE</td><td> 289</td>

<td> 405211</td><td> 1500</td><td> 1511</td><td> CCAGGCCCACAA</td><td> 2-8-2 MOE</td><td> 238</td>

<td> 405212</td><td> 1501</td><td> 1512</td><td> GCCAGGCCCACA</td><td> 2-8-2 MOE</td><td> 239</td>

<td> 379695</td><td> 1643</td><td> 1654</td><td> GGACACCGTCAG</td><td> 1-10-1 MOE</td><td> 266</td>

<td> 379698</td><td> 1875</td><td> 1886</td><td> GCAGCCCACCTC</td><td> 1-10-1 MOE</td><td> 275</td>

<td> 405239</td><td> 1993</td><td> 2004</td><td> GCCAGCATGAGT</td><td> 2-8-2 MOE</td><td> 287</td>

<td> 405211</td><td> 2210</td><td> 2221</td><td> CCAGGCCCACAA</td><td> 2-8-2 MOE</td><td> 238</td>

<td> 405212</td><td> 2211</td><td> 2222</td><td> GCCAGGCCCACA</td><td> 2-8-2 MOE</td><td> 239</td>

In certain embodiments, a target region is nucleotides 85-184 of SEQ ID NO: 3. In certain embodiments, a short antisense compound is targeted to nucleotides 85-184 of SEQ ID NO: 3. In certain such embodiments, a short antisense compound targeted to nucleotides 85-184 comprises a nucleotide 5 sequence selected from SEQ ID NO 214, 215, 216, 217, 218, 219, 221, 222, 223, 224, 225, or 227. In certain such embodiments, a short antisense compound targeted to nucleotides 85-184 of SEQ ID NO: 3 is selected from Isis No 379684, 405193,405194, 405195,405196,405197, 379685,405198, 405199, 405200,405201,379686,379711 or 388628.

In certain embodiments, a target region is nucleotides 113-132 of SEQ ID NO: 3. In certain 10 embodiments, a short antisense compound is targeted to nucleotides 113-132 of SEQ ID NO: 3. In certain such embodiments, a short antisense compound targeted to nucleotides 113-132 comprises a nucleotide sequence selected from SEQ ID NO 215, 216, 217, 218, 219, 221, 222, 223, or 224. In certain such embodiments, a short antisense compound targeted to nucleotides 113-132 of SEQ ID NO: 3 is selected from Isis No 405193,405194,405195,405196,405197,379685,405198,405199,405200, or 405201.

In certain embodiments, a target region is nucleotides 207-329 of SEQ ID NO: 3. In certain

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-73embodiments, a short antisense compound is targeted to nucleotides 207-329 of SEQ ID NO: 3. In certain such embodiments, a short antisense compound targeted to nucleotides 207-329 comprises a nucleotide sequence selected from SEQ ID NO 228,229,230,232,233,234,235,236,237,238,239,240, or 241. In certain such embodiments, a short antisense compound targeted to nucleotides 207-329 of SEQ ID NO: 3 is selected from Isis No 405202,405203,405204,379687,405205,405206,405207,405208,405209, 405210,405211,405212,379688, or 379689.

In certain embodiments, a target region is nucleotides 207-273 of SEQ ID NO: 3. In certain embodiments, a short antisense compound is targeted to nucleotides 207-273 of SEQ ID NO: 3. In certain such embodiments, a short antisense compound targeted to nucleotides 207-273 comprises a nucleotide sequence selected from SEQ ID NO 228, 229,230,232,233,234,235,236,237,238, or 239. In certain such embodiments, a short antisense compound targeted to nucleotides 207-273 of SEQ ID NO: 3 is selected from Isis No 405202, 405203, 405204, 379687, 405205, 405206, 405207, 405208, 405209, 405210,405211, or 405212.

In certain embodiments, a target region is nucleotides 207-219 of SEQ ID NO: 3. In certain embodiments, a short antisense compound is targeted to nucleotides 207-219 of SEQ ID NO: 3. In certain such embodiments, a short antisense compound targeted to nucleotides 207-219 comprises a nucleotide sequence selected from SEQ ID NO 228 or 229. In certain such embodiments, a short antisense compound targeted to nucleotides 207-219 of SEQ ID NO: 3 is selected from Isis NO.. 405202 or 405203.

In certain embodiments, a target region is nucleotides 236-252 of SEQ ID NO: 3. In certain embodiments, a short antisense compound is targeted to nucleotides 236-252 of SEQ ID NO: 3. In certain such embodiments, a short antisense compound targeted to nucleotides 236-252 comprises a nucleotide sequence selected from SEQ ID NO 230,232,233,234,235, or 236. In certain such embodiments, a short antisense compound targeted to nucleotides 236-252 of SEQ ID NO: 3 is selected from Isis NO. 405204, 379687,405205,405206,405207,405208, or 405209.

In certain embodiments, a target region is nucleotides 260-273 of SEQ ID NO: 3. In certain embodiments, a short antisense compound is targeted to nucleotides 260-273 of SEQ ID NO: 3. In certain such embodiments, a short antisense compound targeted to nucleotides 260-273 comprises a nucleotide sequence selected from SEQ ID NO 237, 238, or 239. In certain such embodiments, a short antisense compound targeted to nucleotides 260-273 of SEQ ID NO: 3 is selected from Isis NO. 405210,405211, or 405212.

In certain embodiments, a target region is nucleotides 435-640 of SEQ ID NO: 3. In certain embodiments, a short antisense compound is targeted to nucleotides 435-640 of SEQ ID NO: 3. In certain such embodiments, a short antisense compound targeted to nucleotides 435-640 comprises a nucleotide sequence selected from SEQ ID NO 242,243,245,246,251,252,253,254,256,257,258,259,260,261, 262, 263, or 264. In certain such embodiments, a short antisense compound targeted to nucleotides 435640 of SEQ ID NO: 3 is selected from Isis NO. 379690, 405248, 379691, 389780, 379692, 382676,

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-74388625, 392170, 392173,405213,405214,405215,405216,379693,405217,405218, 405219,405220, 405221,405222,405223,405224, or 379694.

In certain embodiments, a target region is nucleotides 527-540 of SEQ ID NO: 3. In certain embodiments, a short antisense compound is targeted to nucleotides 527-540 of SEQ ID NO: 3. In certain such embodiments, a short antisense compound targeted to nucleotides 527-540 comprises a nucleotide sequence selected from SEQ ID NO 245, 246, or 251. In certain such embodiments, a short antisense compound targeted to nucleotides 527-540 of SEQ ID NO: 3 is selected from Isis NO. 389780, 379692, 382676,388626,392170,392173, or 405213.

In certain embodiments, a target region is nucleotides 564-603 of SEQ ID NO: 3. In certain embodiments, a short antisense compound is targeted to nucleotides 564-603 of SEQ ID NO: 3. In certain such embodiments, a short antisense compound targeted to nucleotides 564-603 comprises a nucleotide sequence selected from SEQ ID NO 252,253,254,256,257,258,259,260,261,262, or 263. In certain such embodiments, a short antisense compound targeted to nucleotides 564-603 of SEQ ID NO: 3 is selected from Isis NO. 405214, 405215, 405216, 379693, 405217, 405218, 405219, 405220, 405221,

405222.405223, or 405224.

In certain embodiments, a target region is nucleotides 564-579 of SEQ ID NO: 3. In certain embodiments, a short antisense compound is targeted to nucleotides 564-579 of SEQ ID NO: 3. In certain such embodiments, a short antisense compound targeted to nucleotides 564-579 comprises a nucleotide sequence selected from SEQ ID NO 252, 253, 254, 256, or 257. In certain such embodiments, a short antisense compound targeted to nucleotides 564-579 of SEQ ID NO: 3 is selected from Isis NO. 405214, 405215,405216,379693,405217, or 405218.

In certain embodiments, a target region is nucleotides 587-603 of SEQ ID NO: 3. In certain embodiments, a short antisense compound is targeted to nucleotides 587-603 of SEQ ID NO: 3. In certain such embodiments, a short antisense compound targeted to nucleotides 587-603 comprises a nucleotide sequence selected from SEQ ID NO 258, 259, 260, 261, 262, or 263. In certain such embodiments, a short antisense compound targeted to nucleotides 587-603 of SEQ ID NO: 3 is selected from Isis NO.

405219.405220.405221.405222.405223, or 405224.

In certain embodiments, a target region is nucleotides 974-1014 of SEQ ID NO: 3. In certain embodiments, a short antisense compound is targeted to nucleotides 974-1014 of SEQ ID NO: 3. In certain such embodiments, a short antisense compound targeted to nucleotides 974-1014 comprises a nucleotide sequence selected from SEQ ID NO 267, 268, 269, 270, 271, 272, or 274. In certain such embodiments, a short antisense compound targeted to nucleotides 974-1014 of SEQ ID NO: 3 is selected from Isis NO. 379696,405226,405227,405228,405229,405230,379697, or 405231.

In certain embodiments, a target region is nucleotides 998-1014 of SEQ ID NO: 3. In certain embodiments, a short antisense compound is targeted to nucleotides 998-1014 of SEQ ID NO: 3. In certain such embodiments, a short antisense compound targeted to nucleotides 998-1014 comprises a nucleotide sequence selected from SEQ ID NO 268, 269, 270, 271, 272, or 274. In certain such

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-75embodiments, a short antisense compound targeted to nucleotides 998-1014 of SEQ ID NO: 3 is selected from Isis NO. 405226,405227,405228,405229,405230,379697, or 405231.

In certain embodiments, a target region is nucleotides 1091-1170 of SEQ ID NO: 3. In certain embodiments, a short antisense compound is targeted to nucleotides 1091-1170 of SEQ ID NO: 3. In certain such embodiments, a short antisense canpound targeted to nucleotides 1091-1170 comprises a nucleotide sequence selected from SEQ ID NO 275,276,277,278,279,280,281,283,284,285,286, or 287. In certain such embodiments, a short antisense compound targeted to nucleotides 1091-1170 of SEQ ID NO: 3 is selected from Isis NO. 379698,405232,405233,405234,405235,388626, 379699, 382677,

405236.405237.405238.379700, or 405239.

In certain embodiments, a target region is nucleotides 1091-1104 of SEQ ID NO: 3. In certain embodiments, a short antisense compound is targeted to nucleotides 1091-1104 of SEQ ID NO: 3. In certain such embodiments, a short antisense compound targeted to nucleotides 1091-1104 comprises a nucleotide sequence selected from SEQ ID NO 275, 276, or 277. In certain such embodiments, an short antisense compound targeted to nucleotides 1091-1104 of SEQ ID NO: 3 is selected from Isis NO. 379698,405232, or 405233.

In certain embodiments, a target region is nucleotides 1130-1144 of SEQ ID NO: 3. In certain embodiments, a short antisense compound is targeted to nucleotides 1130-1144 of SEQ ID NO: 3. In certain such embodiments, a short antisense compound targeted to nucleotides 1130-1144 comprises a nucleotide sequence selected from SEQ ID NO 278,279,280,281, or 283. In certain such embodiments, a short antisense compound targeted to nucleotides 1130-1144 of SEQ ID NO: 3 is selected from Isis NO. 405234,405235,388626,379699,382677, a 405236.

In certain embodiments, a target region is nucleotides 1157-1170 of SEQ ID NO: 3. In certain embodiments, a short antisense compound is targeted to nucleotides 1157-1170 of SEQ ID NO: 3. In certain such embodiments, a short antisense compound targeted to nucleotides 1157-1170 comprises a nucleotide sequence selected from SEQ ID NO 284, 285, or 287. In certain such embodiments, a short antisense compound targeted to nucleotides 1157-1170 of SEQ ID NO: 3 is selected from Isis NO.

405237.405238.379700, or 405239.

In certain embodiments, a target region is nucleotides 1542-1556 of SEQ ID NO: 3. In certain embodiments, a short antisense compound is targeted to nucleotides 1542-1556 of SEQ ID NO: 3. In certain such embodiments, a short antisense compound targeted to nucleotides 1542-1556 comprises a nucleotide sequence selected from SEQ ID NO 289,290,291,292, or 293. In certain such embodiments, a short antisense compound targeted to nucleotides 1542-1556 of SEQ ID NO: 3 is selected from Isis NO. 405240,405241,405242,388629,379702, or 382678.

In certain embodiments, a target region is nucleotides 1976-1991 of SEQ ID NO: 3. In certain embodiments, a short antisense compound is targeted to nucleotides 1976-1991 of SEQ ID NO: 3. In certain such embodiments, a short antisense compound targeted to nucleotides 1976-1991 comprises a nucleotide sequence selected from SEQ ID NO 296,297,298,299, or 300. In certain such embodiments,

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-76a short antisense compound targeted to nucleotides 1976-1991 of SEQ ID NO: 3 is selected from Isis NO. 405243,405244,405245,405246, or 405247.

In certain embodiments, short antisense compounds targeted to an SGLT2 nucleic acid are 8 to 16, preferably 9 to 15, more preferably 9 to 14, more preferably 10 to 14 nucleotides in length. Incertain embodiments, short antisense compounds targeted to an SGLT2 nucleic acid are 9 to 14 nucleotides in length. In certain embodiments, short antisense compounds targeted to an SGLT2 nucleic acid are 10 to 14 nucleotides in length. In certain embodiments, such short antisense compounds are short antisense oligonucleotides.

In certain embodiments, short antisense compounds targeted to an SGLT2 nucleic acid are short gapmers. In certain such embodiments, short gapmers targeted to an SGLT2 nucleic acid comprise at least one high affinity modification in one or more wings of the compound. In certain embodiments, short antisense compounds targeted to an SGLT2 nucleic acid comprise 1 to 3 high-affinity modifications in each wing. In certain such embodiments, the nucleosides or nucleotides of the wing comprise a 2’ modification. In certain such embodiments, the monomers of the wing are BNA’s. In certain such embodiments, the monomers of the wing are selected from Ci-L-Methyleneoxy (4’-CH₂-O-2’) BNA, β-DMethyleneoxy (4’-CH₂-O-2’) BNA , Ethyleneoxy (4’-(CH₂)rO-2’) BNA , Aminooxy (4’-CH₂-O-N(R)2’) BNA and Oxyamino (4’-CH2-N(R)-O-2’) BNA. In certain embodiments, the monomers of a wing comprise a substituent at the 2’ position selected from allyl, amino, azido, thio, O-allyl, O Ci Cio alkyl, OCF₃,0-(CH₂)₂-O-CH,, 2’-O(CH₂)₂SCH3,O-(CH₂)₂-O-N(R_m)(R_fl), and O-CH₂-C(=O)-N(R_o,)(R_n), where each R« and Ro is, independently, H or substituted or unsubstituted Ci-Cio alkyl. In certain embodiments, the monomers of a wing are 2’MOE nucleotides.

In certain embodiments, short antisense compounds targeted to an SGLT2 nucleic acid comprise a gap between the 5’ wing and the 3’ wing. In certain embodiments the gap comprises five, six, seven, eight, nine, ten, eleven, twelve, thirteen, or fourteen monomers. In certain embodiments, the monomers of the gap are unmodified deoxyribonucleotides. In certain embodiments, the monomers of the gap are unmodified ribonucleotides. In certain embodiments, gap modifications (if any) gap result in an antisense compound that, when bound to its target nucleic acid, supports cleavage by an RNase, including, but not limited to, RNase H.

In certain embodiments, short antisense compounds targeted to an SGLT2 nucleic acid have uniform monomeric linkages. In certain such embodiments, those linkages are all phosphorothioate linkages. In certain embodiments, the linkages are all phosphodiester linkages. In certain embodiments, short antisense compounds targeted to an SGLT2 nucleic acid have mixed backbones.

In certain embodiments, short antisense compounds targeted to an SGLT2 nucleic acid are 8 monomers in length. In certain embodiments, short antisense compounds targeted to an SGLT2 nucleic acid are 9 monomers in length. In certain embodiments, short antisense compounds targeted to an SGLT2 nucleic acid are 10 monomers in length. In certain embodiments, short antisense compounds targeted to an SGLT2 nucleic acid are 11 monomers in length. In certain embodiments, short antisense

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-77compounds targeted to an SGLT2 nucleic acid are monomers in length. In certain embodiments, short antisense compounds targeted to an SGLT2 nucleic acid are 13 monomers in length. In certain embodiments, short antisense compounds targeted to an SGLT2 nucleic acid are 14 monomers in length. In certain embodiments, short antisense compounds targeted to an SGLT2 nucleic acid are 15 monomers in length. In certain embodiments, short antisense compounds targeted to an SGLT2 nucleic acid are 16 monomers in length. In certain embodiments, short antisense compounds targeted to an SGLT2 nucleic acid comprise 9 to 15 monomers. In certain embodiments, short antisense compounds targeted to an SGLT2 nucleic acid comprise 10 to 15 monomers. In certain embodiments, short antisense compounds targeted to an SGLT2 nucleic acid comprise 12 to 14 monomers. In certain embodiments, short antisense compounds targeted to an SGLT2 nucleic acid comprise 12 to 14 nucleotides or nucleosides.

In certain embodiments, the invention provides methods of modulating expression of SGLT2. In certain embodiments, such methods comprise use of one or more short antisense compound targeted to an SGLT2 nucleic acid, wherein the short antisense compound targeted to an SGLT2 nucleic acid is from about 8 to about 16, preferably 9 to 15, more preferably 9 to 14, more preferably 10 to 14 monomers (i.e. from about 8 to about 16 linked monomers). One of ordinary skill in the art will appreciate that this comprehends methods of modulating expression of SGLT2 using one or more short antisense compounds targeted to an SGLT2 nucleic acid of 8,9,10,11,12,13,14,15 or 16 monomers.

In certain embodiments, methods of modulating SGLT2 comprise use of a short antisense compound targeted to an SGLT2 nucleic acid that is 8 monomers in length. In certain embodiments, methods of modulating SGLT2 comprise use of a short antisense compound targeted to an SGLT2 nucleic acid that is 9 monomers in length. In certain embodiments, methods of modulating SGLT2 comprise use of a short antisense compound targeted to an SGLT2 nucleic acid that is 10 monomers in length. In certain embodiments, methods of modulating SGLT2 comprise use of a short antisense compound targeted to an SGLT2 nucleic acid that is 11 monomers in length. In certain embodiments, methods of modulating SGLT2 comprise use of a short antisense compound targeted to an SGLT2 nucleic acid that is 12 monomers in length. In certain embodiments, methods of modulating SGLT2 comprise use of a short antisense compound targeted to an SGLT2 nucleic acid that is 13 monomers in length. In certain embodiments, methods of modulating SGLT2 comprise use of a short antisense compound targeted to an SGLT2 nucleic acid that is 14 monomers in length. In certain embodiments, methods of modulating SGLT2 comprise use of a short antisense compound targeted to an SGLT2 nucleic acid that is 15 monomers in length. In certain embodiments, methods of modulating SGLT2 comprise use of a short antisense compound targeted to an SGLT2 nucleic acid that is 16 monomers in length.

In certain embodiments, methods of modulating expression of SGLT2 comprise use of a short antisense compound targeted to an SGLT2 nucleic acid comprising 9 to 15 monomers. In certain embodiments, methods of modulating expression of SGLT2 comprise use of a short antisense compound targeted to an SGLT2 nucleic acid comprising 10 to 15 monomers. In certain embodiments, methods of modulating expression of SGLT2 comprise use of a short antisense compound targeted to an SGLT2

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-78nucleic acid comprising 12 to 14 monomers. In certain embodiments, methods of modulating expression of SGLT2 comprise use of a short antisense compound targeted to an SGLT2 nucleic acid comprising 12 or 14 nucleotides or nucleosides.

3. PCSK9

In individuals with autosomal dominant hypercholesterolemia (ADH), elevated LDL-C levels have been linked to mutations in the genes encoding LDL-receptor (LDL-R), apolipoprotein B (apoB), or proprotein convertase subtilisin/kexin type 9 (PCSK.9) (Abifadel et al., Nat. Genet., 2003,34:154-156). PCSK9 was identified as a third locus associated with ADH when gain-of-function mutations in PCSK.9 were found to be linked to elevated LDL-C levels. ApoB participates in the intracellular assembly and secretion of triglyceride-rich lipoproteins and is a ligand for the LDL-R. PCSK9 is proposed to reduce LDL-R expression levels in the liver. Reduced LDL-R expression results in reduced hepatic uptake of circulating ApoB-containing lipoproteins, which in tum leads to elevated cholesterol.

Definitions “PCSK9” is the gene product or protein of which expression is to be modulated by administration of a short antisense compound.

“PCSK9 nucleic acid means any nucleic acid encoding PCSK9. For example, in certain embodiments, a PCSK9nucleic acid includes, without limitation, a DNA sequence encoding PCSK9, an RNA sequence transcribed from DNA encoding PCSK9, and an mRNA sequence encoding PCSK9.

“PCSK9 mRNA” means an mRNA encoding PCSK9.

PCSK9 Therapeutic Indications

In certain embodiments, the invention provides methods of modulating the expression of PCSK9 in an individual comprising administering a short antisense compound targeted to a PCSK9 nucleic acid. In certain embodiments, the invention provides methods of treating an individual comprising administering one or more pharmaceutical compositions of the present invention. In certain embodiments, the individual has hypercholesterolemia, mixed dyslipidemia, atherosclerosis, a risk of developing atherosclerosis, coronary heart disease, a history of coronary heart disease, early onset coronary heart disease, one or more risk factors for coronary heart disease, type II diabetes, type II diabetes with dyslipidemia, dyslipidemia, hypertriglyceridemia, hyperlipidemia, hyperfattyacidemia, hepatic steatosis, non-alcoholic steatohepatitis, or non-alcoholic fatty liver disease.

Guidelines for lipid-lowering therapy were established in 2001 by Adult Treatment Panel III (ATP III) of die National Cholesterol Education Program (NCEP), and updated in 2004 (Grundy et al., Circulation, 2004, 110, 227-239). The guidelines include obtaining a complete lipoprotein profile, typically after a 9 to 12 hour fast, for determination of LDL-C, total cholesterol, and HDL-C levels. According to the most recently established guidelines, LDL-C levels of 130-159 mg/dL, 160-189 mg/dL,

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-79and greater than or equal to 190 mg/dL are considered borderline high, high, and very high, respectively. Total cholesterol levels of 200-239 and greater than or equal to 240 mg/dL are considered borderline high and high, respectively. HDL-C levels of less than 40 mg/dL are considered low.

In certain embodiments, the individual has been identified as in need of lipid-lowering therapy. In certain such embodiments, the individual has been identified as in need of lipid-lowering therapy according to the guidelines established in 2001 by Adult Treatment Panel III (ATP III) of the National Cholesterol Education Program (NCEP), and updated in 2004 (Grundy et al., Circulation, 2004, 110,227239). In certain such embodiments, the individual in need of lipid-lowering therapy has LDL-C above 190 mg/dL. In certain such embodiments, the individual in need of lipid-lowering therapy has LDL-C above 160 mg/dL. In certain such embodiments, the individual in need of lipid-lowering therapy has LDL-C above 130 mg/dL. In certain such embodiments the individual in need of lipid-lowering therapy has LDLC above 100 mg/dL. In certain such embodiments the individual in need of lipid-lowering therapy should maintain LDL-C below 160 mg/dL. In certain such embodiments the individual in need of lipid-lowering therapy should maintain LDL-C below 130 mg/dL. In certain such embodiments the individual in need of lipid-lowering therapy should maintain LDL-C below 100 mg/dL. In certain such embodiments the individual should maintain LDL-C below 70 mg/dL.

In certain embodiments the invention provides methods for reducing ApoB in an individual. In certain embodiments the invention provides methods for reducing ApoB-containing lipoprotein in an individual. In certain embodiments the invention provides methods for reducing LDL-C in an individual. In certain embodiments the invention provides methods for reducing VLDL-C in an individual. In certain embodiments the invention provides methods for reducing IDL-C in an individual. In certain embodiments the invention provides methods for reducing non-HDL-C in an individual. In certain embodiments the invention provides methods for reducing Lp(a) in an individual. In certain embodiments the invention provides methods for reducing serum triglyceride in an individual. In certain embodiments the invention provides methods for reducing liver triglyceride in an individual. In certain embodiments the invention provides methods for reducing Ox-LDL-C in an individual. In certain embodiments the invention provides methods for reducing small LDL particles in an individual. In certain embodiments the invention provides methods for reducing small VLDL particles in an individual. In certain embodiments the invention provides methods for reducing phospholipids in an individual. In certain embodiments the invention provides methods for reducing oxidized phospholipids in an individual.

In certain embodiments, the methods provided by the present invention do not lower HDL-C. In certain embodiments, the methods provided by the present invention do not result in accumulation of lipids in the liver.

In certain embodiments a pharmaceutical composition comprising a short antisense compound targeted to a PCSK9 nucleic acid is for use in therapy. In certain embodiments, the therapy is the reduction of LDL-C, ApoB, VLDL-C, IDL-C, non-HDL-C, Lp(a), serum triglyceride, liver triglyceride, Ox-LDL-C, small LDL particles, small VLDL, phospholipids, or oxidized phospholipids in an individual.

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-80In certain embodiments, the therapy is the treatment of hypercholesterolemia, mixed dyslipidemia, atherosclerosis, a risk of developing atherosclerosis, coronaiy heart disease, a history of coronary heart disease, early onset coronary heart disease, one or more risk factors for coronary heart disease, type II diabetes, type II diabetes with dyslipidemia, dyslipidemia, hypertriglyceridemia, hyperlipidemia, hyperfattyacidemia, hepatic steatosis, non-alcoholic steatohepatitis, or non-alcoholic fatty liver disease. In additional embodiments, the therapy is the reduction of CHD risk. In certain the therapy is prevention of atherosclerosis. In certain embodiments, the therapy is the prevention of coronary heart disease.

In certain embodiments a pharmaceutical composition comprising a short antisense compound targeted to a PCSK9 nucleic acid is used for the preparation of a medicament for reducing LDL-C, ApoB, VLDL-C, IDL-C, non-HDL-C, Lp(a), serum triglyceride, liver triglyceride, Ox-LDL-C, small LDL particles, small VLDL, phospholipids, or oxidized phospholipids in an individual. In certain embodiments pharmaceutical composition comprising a short antisense compound targeted to PCKS9 is used for the preparation of a medicament for reducing coronary heart disease risk. In certain embodiments a short antisense compound targeted to a PCSK9 nucleic acid is used for the preparation of a medicament for the treatment of hypercholesterolemia, mixed dyslipidemia, atherosclerosis, a risk of developing atherosclerosis, coronary heart disease, a history of coronary heart disease, early onset coronary heart disease, one or more risk factors for coronary heart disease, type II diabetes, type II diabetes with dyslipidemia, dyslipidemia, hypertriglyceridemia, hyperlipidemia, hyperfattyacidemia, hepatic steatosis, non-alcoholic steatohepatitis, or non-alcoholic fatty liver disease.

PCSK9 Combination Therapies

In certain embodiments, one or more pharmaceutical compositions of the present invention are co-administered with one or more other pharmaceutical agents. In certain embodiments, such one or more other pharmaceutical agents are designed to treat the same disease or condition as the one or more pharmaceutical compositions of the present invention. In certain embodiments, such one or more other pharmaceutical agents are designed to treat a different disease or condition as the one or more pharmaceutical compositions of the present invention. In certain embodiments, such one or more other pharmaceutical agents are designed to treat an undesired effect of one or more pharmaceutical compositions of the present invention. In certain embodiments, one or more pharmaceutical compositions of the present invention are co-administered with another pharmaceutical agent to treat an undesired effect of that other pharmaceutical agent. In certain embodiments, one or more pharmaceutical compositions of the present invention and one or more other pharmaceutical agents are administered at the same time. In certain embodiments, one or more pharmaceutical compositions of the present invention and one or more other pharmaceutical agents are administered at different times. In certain embodiments, one or more pharmaceutical compositions of the present invention and one or more other pharmaceutical agents are prepared together in a single formulation. In certain embodiments, one or more pharmaceutical compositions of the present invention and one or more other pharmaceutical agents are prepared

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-81separately.

In certain embodiments, pharmaceutical agents that may be co-administered with a pharmaceutical composition of the present invention include lipid-lowering agents. In certain such embodiments, pharmaceutical agents that may be co-administered with a pharmaceutical composition of the present invention include, but are not limited to atorvastatin, simvastatin, rosuvastatin, and ezetimibe. In certain such embodiments, the lipid-lowering agent is administered prior to administration of a pharmaceutical composition of the present invention. In certain such embodiments, the lipid-lowering agent is administered following administration of a pharmaceutical composition of the present invention. In certain such embodiments the lipid-lowering agent is administered at the same time as a pharmaceutical composition of the present invention. In certain such embodiments the dose of a coadministered lipid-lowering agent is the same as the dose that would be administered if the lipid-lowering agent was administered alone. In certain such embodiments the dose of a co-administered lipid-lowering agent is lower than the dose that would be administered if the lipid-lowering agent was administered alone. In certain such embodiments the dose of a co-administered lipid-lowering agent is greater than the dose that would be administered if the lipid-lowering agent was administered alone.

In certain embodiments, a co-administered lipid-lowering agent is a HMG-CoA reductase inhibitor. In certain such embodiments the HMG-CoA reductase inhibitor is a statin. In certain such embodiments the statin is selected from atorvastatin, simvastatin, pravastatin, fluvastatin, and rosuvastatin.

In certain embodiments, a co-administered lipid-lowering agent is a cholesterol absorption inhibitor. In certain such embodiments, cholesterol absorption inhibitor is ezetimibe.

In certain embodiments, a co-administered lipid-lowering agent is a co-formulated HMG-CoA reductase inhibitor and cholesterol absorption inhibitor. In certain such embodiments the co-formulated lipid-lowering agent is ezetimibe/simvastatin.

In certain embodiments, a co-administered lipid-lowering agent is a microsomal triglyceride transfer protein inhibitor (MTP inhibitor).

In certain embodiments, a co-administered lipid-lowering agent is an oligonucleotide targeted to an ApoB nucleic acid.

In certain embodiments, a co-administered pharmaceutical agent is a bile acid séquestrant. In certain such embodiments, the bile acid séquestrant is selected from cholestyramine, colestipol, and colesevelam.

In certain embodiments, a co-administered pharmaceutical agent is a nicotinic acid. In certain such embodiments, the nicotinic acid is selected from immediate release nicotinic acid, extended release nicotinic acid, and sustained release nicotinic acid.

In certain embodiments, a co-administered pharmaceutical agent is a fibric acid. In certain such embodiments, a fibric acid is selected from gemfibrozil, fenofibrate, clofibrate, bezafibrate, and ciprofibrate.

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-82Furthcr examples of pharmaceutical agents that may be co-administered with a pharmaceutical composition of the present invention include, but are not limited to, corticosteroids, including but not limited to prednisone; immunoglobulins, including, but not limited to intravenous immunoglobulin (IVIg); analgesics (e.g., acetaminophen); anti-inflammatory agents, including, but not limited to nonsteroidal anti-inflammatory drugs (e.g., ibuprofen, COX-1 inhibitors, and COX-2, inhibitors); salicylates; antibiotics; antivirals; antifungal agents; antidiabetic agents (e.g., biguanides, glucosidase inhibitors, insulins, sulfonylureas, and thiazolidenediones); adrenergic modifiers; diuretics; hormones (e.g., anabolic steroids, androgen, estrogen, calcitonin, progestin, somatostan, and thyroid hormones); immunomodulators; muscle relaxants; antihistamines; osteoporosis agents (e.g., biphosphonates, calcitonin, and estrogens); prostaglandins, antineoplastic agents; psychotherapeutic agents; sedatives; poison oak or poison sumac products; antibodies; and vaccines.

In certain embodiments, the pharmaceutical compositions of the present invention may be administered in conjuction with a lipid-lowering therapy. In certain such embodiments, a lipid-lowering therapy is therapeutic lifestyle change. In certain such embodiments, a lipid-lowering therapy is LDL apheresis.

Certain Short Antisense Compounds Targeted to a PCSK9 Nucleic Acid

In certain embodiments, short antisense compounds are targeted to a PCSK9 nucleic acid having the sequence of GENBANK® Accession No. NM_174936.2, incorporated herein as SEQ ID NO: 4. In certain such embodiments, a short antisense compound targeted to SEQ ID NO: 4 is at least 90% complementary to SEQ ID NO: 4. In certain such embodiments, a short antisense compound targeted to SEQ ID NO: 4 is at least 95% complementary to SEQ ID NO: 4. In certain such embodiments, a short antisense compound targeted to SEQ ID NO: 4 is 100% complementary to SEQ ID NO: 4. In certain embodiments, a short antisense compound targeted to SEQ ID NO: 4 comprises a nucleotide sequence selected from the nucleotide sequences set forth in Table 6 or Table 7.

The nucleotide sequence set forth in each SEQ ID NO in Tables 6 and 7 is independent of any modification to a sugar moiety, an intemucleoside linkage, or a nucleobase. As such, short antisense compounds defined by a SEQ ID NO may comprise, independently, one or more modifications to a sugar moiety, an intemucleoside linkage, or a nucleobase. Short antisense compounds described by Isis Number (Isis NO.) indicate a combination of nucleobase sequence and one or more modifications to a sugar moiety, an intemucleoside linkage, or a nucleobase.

Tables 6 and 7 illustrate examples of short antisense compounds targeted to SEQ ID NO: 4. Table 6 illustrates short antisense compounds that are 100% complementary to SEQ ID NO: 4. Table 7 illustrates short antisense compounds that have one or two mismatches with respect to SEQ ID NO: 4. The column labeled ‘gapmer motif indicates the wing-gap-wing motif of each short antisense compounds. The gap segment comprises 2*-deoxynucleotides and each nucleotide of each wing segment comprises a 2’-modified sugar. The particular 2’-modified sugar is also indicated in the ‘gapmer motif

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-83column. For example, ‘2-10-2 MOE’ means a 2-10-2 gapmer motif, where a gap segment of ten 2’deoxynucleotides is flanked by wing segments of two nucleotides, where the nucleotides of the wing segments are 2’-MOE nucleotides. Intemucleoside linkages are phosphorothioate. The short antisense compounds comprise 5-methylcytidine in place of unmodified cytosine, unless “unmodified cytosine” is 5 listed in the gapmer motif column, in which case the indicated cytosines are unmodified cytosines. For example, “5-mC in gap only” indicates that the gap segment has 5-methylcytosines, while the wing segments have unmodified cytosines.

Table 6: Short Antisense Compounds targeted to SEQ ID NO: 4

<td> ISIS NO.</td><td> 5' Target Site</td><td> 3’ Target Site</td><td> Sequence (5’-3’)</td><td> Gapmer Motif</td><td> SEQ ID NO</td>

<td> 400297</td><td> 695</td><td> 708</td><td> ATGGGGCAACTTCA</td><td> 2-10-2 MOE</td><td> 329</td>

<td> 400298</td><td> 696</td><td> 709</td><td> CATGGGGCAACTTC</td><td> 2-10-2 MOE</td><td> 330</td>

<td> 400299</td><td> 697</td><td> 710</td><td> ACATGGGGCAACTT</td><td> 2-10-2 MOE</td><td> 331</td>

<td> 400300</td><td> 742</td><td> 755</td><td> GGGATGCTCTGGGC</td><td> 2-10-2 MOE</td><td> 332</td>

<td> 400301</td><td> 757</td><td> 770</td><td> CGCTCCAGGTTCCA</td><td> 2-10-2 MOE</td><td> 333</td>

<td> 400302</td><td> 828</td><td> 841</td><td> GATACACCTCCACC</td><td> 2-10-2 MOE</td><td> 334</td>

<td> 400303</td><td> 829</td><td> 842</td><td> AGATACACCTCCAC</td><td> 2-10-2 MOE</td><td> 335</td>

<td> 400304</td><td> 830</td><td> 843</td><td> GAGATACACCTCCA</td><td> 2-10-2 MOE</td><td> 336</td>

<td> 400305</td><td> 937</td><td> 950</td><td> GCCTGTCTGTGGAA</td><td> 2-10-2 MOE</td><td> 337</td>

<td> 400306</td><td> 952</td><td> 965</td><td> CTGTCACACTTGCT</td><td> 2-10-2 MOE</td><td> 338</td>

<td> 400307</td><td> 988</td><td> 1001</td><td> CGGCCGCTGACCAC</td><td> 2-10-2 MOE</td><td> 339</td>

<td> 400308</td><td> 989</td><td> 1002</td><td> CCGGCCGCTGACCA</td><td> 2-10-2 MOE</td><td> 340</td>

<td> 400309</td><td> 990</td><td> 1003</td><td> CCCGGCCGCTGACC</td><td> 2-10-2 MOE</td><td> 341</td>

<td> 400310</td><td> 991</td><td> 1004</td><td> TCCCGGCCGCTGAC</td><td> 2-10-2 MOE</td><td> 342</td>

<td> 400311</td><td> 992</td><td> 1005</td><td> ATCCCGGCCGCTGA</td><td> 2-10-2 MOE</td><td> 343</td>

<td> 400312</td><td> 993</td><td> 1006</td><td> CATCCCGGCCGCTG</td><td> 2-10-2 MOE</td><td> 344</td>

<td> 400313</td><td> 994</td><td> 1007</td><td> GCATCCCGGCCGCT</td><td> 2-10-2 MOE</td><td> 345</td>

<td> 400314</td><td> 1057</td><td> 1070</td><td> GTGCCCTTCCCTTG</td><td> 2-10-2 MOE</td><td> 346</td>

<td> 400315</td><td> 1075</td><td> 1088</td><td> ATGAGGGTGCCGCT</td><td> 2-10-2 MOE</td><td> 347</td>

<td> 400316</td><td> 1076</td><td> 1089</td><td> TATGAGGGTGCCGC</td><td> 2-10-2 MOE</td><td> 348</td>

<td> 400317</td><td> 1077</td><td> 1090</td><td> CTATGAGGGTGCCG</td><td> 2-10-2 MOE</td><td> 349</td>

<td> 400318</td><td> 1078</td><td> 1091</td><td> CCTATGAGGGTGCC</td><td> 2-10-2 MOE</td><td> 350</td>

<td> 400319</td><td> 1093</td><td> 1106</td><td> CGAATAAACTCCAG</td><td> 2-10-2 MOE</td><td> 351</td>

<td> 400320</td><td> 1094</td><td> 1107</td><td> CCGAATAAACTCCA</td><td> 2-10-2 MOE</td><td> 352</td>

<td> 400321</td><td> 1095</td><td> 1108</td><td> TCCGAATAAACTCC</td><td> 2-10-2 MOE</td><td> 353</td>

<td> 400322</td><td> 1096</td><td> 1109</td><td> TTCCGAATAAACTC</td><td> 2-10-2 MOE</td><td> 354</td>

<td> 400323</td><td> 1147</td><td> 1160</td><td> GCCAGGGGCAGCAG</td><td> 2-10-2 MOE</td><td> 355</td>

<td> 400324</td><td> 1255</td><td> 1268</td><td> GAGTAGAGGCAGGC</td><td> 2-10-2 MOE</td><td> 356</td>

<td> 400325</td><td> 1334</td><td> 1347</td><td> CCCCAAAGTCCCCA</td><td> 2-10-2 MOE</td><td> 357</td>

<td> 400326</td><td> 1335</td><td> 1348</td><td> TCCCCAAAGTCCCC</td><td> 2-10-2 MOE</td><td> 358</td>

<td> 400327</td><td> 1336</td><td> 1349</td><td> GTCCCCAAAGTCCC</td><td> 2-10-2 MOE</td><td> 359</td>

<td> 400328</td><td> 1453</td><td> 1466</td><td> ACGTGGGCAGCAGC</td><td> 2-10-2 MOE</td><td> 360</td>

<td> 400329</td><td> 1454</td><td> 1467</td><td> CACGTGGGCAGCAG</td><td> 2-10-2 MOE</td><td> 361</td>

<td> 400330</td><td> 1455</td><td> 1468</td><td> CCACGTGGGCAGCA</td><td> 2-10-2 MOE</td><td> 362</td>

<td> 400331</td><td> 1456</td><td> 1469</td><td> GCCACGTGGGCAGC</td><td> 2-10-2 MOE</td><td> 363</td>

<td> 400332</td><td> 1569</td><td> 1582</td><td> CAGGGAACCAGGCC</td><td> 2-10-2 MOE</td><td> 364</td>

<td> 400333</td><td> 1570</td><td> 1583</td><td> TCAGGGAACCAGGC</td><td> 2-10-2 MOE</td><td> 365</td>

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<td> 400334</td><td> 1571</td><td> 1584</td><td> CTCAGGGAACCAGG</td><td> 2-10-2 MOE</td><td> 366</td>

<td> 400335</td><td> 1572</td><td> 1585</td><td> CCTCAGGGAACCAG</td><td> 2-10-2 MOE</td><td> 367</td>

<td> 400336</td><td> 1573</td><td> 1586</td><td> TCCTCAGGGAACCA</td><td> 2-10-2 MOE</td><td> 368</td>

<td> 400337</td><td> 1574</td><td> 1587</td><td> GTCCTCAGGGAACC</td><td> 2-10-2 MOE</td><td> 369</td>

<td> 400338</td><td> 1575</td><td> 1588</td><td> GGTCCTCAGGGAAC</td><td> 2-10-2 MOE</td><td> 370</td>

<td> 400339</td><td> 1576</td><td> 1589</td><td> TGGTCCTCAGGGAA</td><td> 2-10-2 MOE</td><td> 371</td>

<td> 400340</td><td> 1577</td><td> 1590</td><td> CTGGTCCTCAGGGA</td><td> 2-10-2 MOE</td><td> 372</td>

<td> 400341</td><td> 1578</td><td> 1591</td><td> GCTGGTCCTCAGGG</td><td> 2-10-2 MOE</td><td> 373</td>

<td> 400342</td><td> 1621</td><td> 1634</td><td> GTGCTGGGGGGCAG</td><td> 2-10-2 MOE</td><td> 374</td>

<td> 400343</td><td> 1622</td><td> 1635</td><td> GGTGCTGGGGGGCA</td><td> 2-10-2 MOE</td><td> 375</td>

<td> 400344</td><td> 1623</td><td> 1636</td><td> GGGTGCTGGGGGGC</td><td> 2-10-2 MOE</td><td> 376</td>

<td> 400345</td><td> 1624</td><td> 1637</td><td> TGGGTGCTGGGGGG</td><td> 2-10-2 MOE</td><td> 377</td>

<td> 400346</td><td> 1738</td><td> 1751</td><td> GÂGCAGCTCAGCAG</td><td> 2-10-2 MOE</td><td> 378</td>

<td> 400347</td><td> 1739</td><td> 1752</td><td> GGAGCAGCTCAGCA</td><td> 2-10-2 MOE</td><td> 379</td>

<td> 400348</td><td> 1740</td><td> 1753</td><td> TGGAGCAGCTCAGC</td><td> 2-10-2 MOE</td><td> 380</td>

<td> 400349</td><td> 1741</td><td> 1754</td><td> CTGGAGCAGCTCAG</td><td> 2-10-2 MOE</td><td> 381</td>

<td> 400350</td><td> 1834</td><td> 1847</td><td> CCCTCACCCCCAAA</td><td> 2-10-2 MOE</td><td> 382</td>

<td> 400351</td><td> 1835</td><td> 1848</td><td> ACCCTCACCCCCAA</td><td> 2-10-2 MOE</td><td> 383</td>

<td> 400352</td><td> 1836</td><td> 1849</td><td> CACCCTCACCCCCA</td><td> 2-10-2 MOE</td><td> 384</td>

<td> 400353</td><td> 1837</td><td> 1850</td><td> ACACCCTCACCCCC</td><td> 2-10-2 MOE</td><td> 385</td>

<td> 400354</td><td> 1838</td><td> 1851</td><td> GACACCCTCACCCC</td><td> 2-10-2 MOE</td><td> 386</td>

<td> 400355</td><td> 1839</td><td> 1852</td><td> AGACACCCTCACCC</td><td> 2-10-2 MOE</td><td> 387</td>

<td> 400356</td><td> 1840</td><td> 1853</td><td> TAGACACCCTCACC</td><td> 2-10-2 MOE</td><td> 388</td>

<td> 400357</td><td> 2083</td><td> 2096</td><td> TGGCAGCAGGAAGC</td><td> 2-10-2 MOE</td><td> 389</td>

<td> 400358</td><td> 2084</td><td> 2097</td><td> ATGGCAGCAGGAAG</td><td> 2-10-2 MOE</td><td> 390</td>

<td> 400359</td><td> 2085</td><td> 2098</td><td> CATGGCAGCAGGAA</td><td> 2-10-2 MOE</td><td> 391</td>

<td> 400360</td><td> 2086</td><td> 2099</td><td> GCATGGCAGCAGGA</td><td> 2-10-2 MOE</td><td> 392</td>

<td> 400361</td><td> 2316</td><td> 2329</td><td> GGCAGCAGATGGCA</td><td> 2-10-2 MOE</td><td> 393</td>

<td> 400362</td><td> 2317</td><td> 2330</td><td> CGGCAGCAGATGGC</td><td> 2-10-2 MOE</td><td> 394</td>

<td> 400363</td><td> 2318</td><td> 2331</td><td> CCGGCAGCAGATGG</td><td> 2-10-2 MOE</td><td> 395</td>

<td> 400364</td><td> 2319</td><td> 2332</td><td> TCCGGCAGCAGATG</td><td> 2-10-2 MOE</td><td> 396</td>

<td> 400365</td><td> 2320</td><td> 2333</td><td> CTCCGGCAGCAGAT</td><td> 2-10-2 MOE</td><td> 397</td>

<td> 400366</td><td> 2321</td><td> 2334</td><td> GCTCCGGCAGCAGA</td><td> 2-10-2 MOE</td><td> 398</td>

<td> 400367</td><td> 2322</td><td> 2335</td><td> GGCTCCGGCAGCAG</td><td> 2-10-2 MOE</td><td> 399</td>

<td> 400368</td><td> 2323</td><td> 2336</td><td> CGGCTCCGGCAGCA</td><td> 2-10-2 MOE</td><td> 400</td>

<td> 400369</td><td> 2324</td><td> 2337</td><td> CCGGCTCCGGCAGC</td><td> 2-10-2 MOE</td><td> 401</td>

<td> 400370</td><td> 2325</td><td> 2338</td><td> GCCGGCTCCGGCAG</td><td> 2-10 2 MOE</td><td> 402</td>

<td> 400371</td><td> 3543</td><td> 3556</td><td> AGTTACAAAAGCAA</td><td> 2-10-2 MOE</td><td> 403</td>

<td> 403739</td><td> 988</td><td> 1001</td><td> CGGCCGCTGACCAC</td><td> 2-10-2 (6'S)-6'-methylMethyleneoxy BNA</td><td> 339</td>

<td> 403740</td><td> 1455</td><td> 1468</td><td> CCACGTGGGCAGCA</td><td> 2-10-2 (6^,S)-6'-methylMethyleneoxy BNA</td><td> 362</td>

Table 7: Short antisense compounds targeted to SEQ ID NO: 4 and having 1 or 2 mismatches

<td> ISIS NO.</td><td> 5' Target Site</td><td> 3' Target Site</td><td> Sequence (5’-3’)</td><td> Gapmer Motif</td><td> SEQ Π) NO</td>

<td> 400323</td><td> 349</td><td> 362</td><td> GCCAGGGGCAGCAG</td><td> 2-10-2 MOE</td><td> 355</td>

<td> 400370</td><td> 679</td><td> 692</td><td> GCCGGCTCCGGCAG</td><td> 2-10-2 MOE</td><td> 402</td>

<td> 400361</td><td> 1860</td><td> 1873</td><td> GGCAGCAGATGGCA</td><td> 2-10-2 MOE</td><td> 393</td>

<td> 400323</td><td> 1873</td><td> 1886</td><td> GCCAGGGGCAGCAG</td><td> 2-10-2 MOE</td><td> 355</td>

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<td> 400310</td><td> 2257</td><td> 2270</td><td> TCCCGGCCGCTGAC</td><td> 2-10-2 MOE</td><td> 342</td>

<td> 400361</td><td> 2653</td><td> 2666</td><td> GGCAGCAGATGGCA</td><td> 2-10-2 MOE</td><td> 393</td>

<td> 400350</td><td> 2811</td><td> 2824</td><td> CCCTCACCCCCAAA</td><td> 2-10-2 MOE</td><td> 382</td>

<td> 400351</td><td> 2812</td><td> 2825</td><td> ACCCTCACCCCCAA</td><td> 2-10-2 MOE</td><td> 383</td>

<td> 400352</td><td> 2813</td><td> 2826</td><td> CACCCTCACCCCCA</td><td> 2-10-2 MOE</td><td> 384</td>

<td> 400353</td><td> 2814</td><td> 2827</td><td> ACACCCTCACCCCC</td><td> 2-10-2 MOE</td><td> 385</td>

<td> 400334</td><td> 2966</td><td> 2979</td><td> CTCAGGGAACCAGG</td><td> 2-10-2 MOE</td><td> 366</td>

<td> 400332</td><td> 3379</td><td> 3392</td><td> CAGGGAACCAGGCC</td><td> 2-10-2 MOE</td><td> 364</td>

<td> 400340</td><td> 3448</td><td> 3461</td><td> CTGGTCCTCAGGGA</td><td> 2-10-2 MOE</td><td> 372</td>

<td> 400341</td><td> 3449</td><td> 3462</td><td> GCTGGTCCTCAGGG</td><td> 2-10-2 MOE</td><td> 373</td>

In certain embodiments, a target region is nucleotides 695-710 of SEQ ID NO: 4. In certain such embodiments, short antisense compounds targeted to nucleotides 695-710 of SEQ ID NO: 4 comprise a nucleotide sequence selected from SEQ ID NO: 329, 330, or 331. In certain such embodiments, a short antisense compound targeted to nucleotides 695-710 of SEQ ID NO: 4 is selected from Isis NO. 400297, 400298, or 400299.

In certain embodiments, a target region is nucleotides 742-770 of SEQ ID NO: 4. In certain such embodiments, a short antisense compound targeted to nucleotides 742-770 of SEQ ID NO: 4 comprises a nucleotide sequence selected from SEQ ID NO 332 or 333. In certain such embodiments, a short antisense compound targeted to nucleotides 742-770 of SEQ ID NO: 4 is selected from Isis NO. 400300 or 400301.

In certain embodiments, a target region is nucleotides 828-843 of SEQ ID NO: 4. In certain such embodiments, a short antisense compound targeted to nucleotides 828-843 of SEQ ID NO: 4 comprises a nucleotide sequence selected from SEQ ID NO 334, 335, or 336. In certain such embodiments, a short antisense compound targeted to nucleotides 828-843 of SEQ ID NO: 4 is selected from ISIS No. 400302, 400303, or 400304.

In certain embodiments, a target region is nucleotides 937-1007 of SEQ ID NO: 4. In certain such embodiments, a short antisense compound targeted to nucleotides 937-1007 of SEQ ID NO: 4 comprises a nucleotide sequence selected from SEQ ID NO 337, 338, 339, 340, 341, 342, 343, 344, or 345. In certain such embodiments, a short antisense compound targeted to nucleotides 937-1007 of SEQ ID NO: 4 is selected from Isis NO. 400305,400306,400307,400308,400309,400310,400311,400312,400313, or 403739.

In certain embodiments, a target region is nucleotides 937-965 of SEQ ID NO: 4. In certain such embodiments, a short antisense compound targeted to nucleotides 937-965 of SEQ ID NO: 4 comprises a nucleotide sequence selected from SEQ ID NO 337 or 338. In certain such embodiments, a short antisense compound targeted to nucleotides 937-965 of SEQ ID NO: 4 is selected from Isis NO. 400305 or 400306.

In certain embodiments, a target region is nucleotides 988-1007 of SEQ ID NO: 4. In certain such embodiments, a short antisense compound targeted to nucleotides 988-1007 of SEQ ID NO: 4 comprises a nucleotide sequence selected from SEQ ID NO 339, 340, 341, 342, 343, 344, or 345. In certain such

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-86embodiments, a short antisense compound targeted to nucleotides 937-1007 of SEQ ID NO: 4 is selected from Isis NO. 400307,400308,400309,400310,400311,400312,4003313, or 403739.

In certain embodiments, a target region is nucleotides 1057-1160 of SEQ ID NO: 4. In certain such embodiments, a short antisense compound targeted to nucleotides 1057-1160 of SEQ ID NO: 4 comprises a nucleotide sequence selected from SEQ ID NO 346,347, 348, 349, 350,351, 352,353,354, or 355. In certain such embodiments, a short antisense compound targeted to nucleotides 1057-1160 of SEQ ID NO: 4 is selected from ISIS NO. 400314, 400315,400316, 400317, 400318, 400319, 400320, 400321,400322, or 400323.

In certain embodiments, a target region is nucleotides 1057-1109 of SEQ ID NO: 4. In certain such embodiments, a short antisense compound targeted to nucleotides 1057-1109 of SEQ ID NO: 4 comprises a nucleotide sequence selected from SEQ ID NO 346, 347, 348, 349, 350, 351, 352, 353, or 354. In certain such embodiments, a short antisense compound targeted to nucleotides 1057-1109 of SEQ ID NO: 4 is selected from ISIS NO. 400314,400315,400316,400317,400318,400319,400320,400321, or 400322.

In certain embodiments, a target region is nucleotides 1057-1091 of SEQ ID NO: 4. In certain such embodiments, a short antisense compound targeted to nucleotides 1057-1091 of SEQ ID NO: 4 comprises a nucleotide sequence selected from SEQ ID NO 346, 347, 348, 349, or 350. In certain such embodiments, a short antisense compound targeted to nucleotides 1057-1091 of SEQ ID NO: 4 is selected from ISIS NO. 400314,400315,400316,400317, or 400318.

In certain embodiments, a target region is nucleotides 1093-1109 of SEQ ID NO: 4. In certain such embodiments, a short antisense compound targeted to nucleotides 1093-1109 of SEQ ID NO: 4 comprises a nucleotide sequence selected from SEQ ID NO 351, 352, 353, or 354. In certain such embodiments, a short antisense compound targeted to nucleotides 1057-1109 of SEQ ID NO: 4 is selected from ISIS NO. 400319,400320,400321, or 400322.

In certain embodiments, a target region is nucleotides 1334-1349 of SEQ ID NO: 4. In certain such embodiments, a short antisense compound targeted to nucleotides 1334-1349 of SEQ ID NO: 4 comprises a nucleotide sequence selected from SEQ ID NO 357, 358, or 359. In certain such embodiments, a short antisense compound targeted to nucleotides 1334-1349 of SEQ ID NO: 4 is selected from ISIS NO 400325,400326, or 400327.

In certain embodiments, a target region is nucleotides 1453-1469 of SEQ ID NO: 4. In certain such embodiments, a short antisense compound targeted to nucleotides 1453-1469 of SEQ ID NO: 4 comprises a nucleotide sequence selected from SEQ ID NO 360, 361, 362, or 363. In certain such embodiments, a short antisense compound targeted to nucleotides 1453-1469 of SEQ ID NO: 4 is selected from ISIS NO 400328,400329,400330,400331, or 403470.

In certain embodiments, a target region is nucleotides 1569-1591 of SEQ ID NO: 4. In certain such embodiments, a short antisense compound targeted to nucleotides 1569-1591 of SEQ ID NO: 4 comprises a nucleotide sequence selected from SEQ ID NO 364, 365,366, 367, 368,369,370, 371,372,

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-87or 373. In certain such embodiments, a short antisense compound targeted to nucleotides 1569-1591 of SEQ ID NO: 4 is selected from ISIS NO 400332, 400333, 400334, 400335, 400336, 400337, 400338, 400339,400340, or 400341.

In certain embodiments, a target region is nucleotides 1621-1637 of SEQ ID NO: 4. In certain such embodiments, a short antisense compound targeted to nucleotides 1621-1637 of SEQ ID NO: 4 comprises a nucleotide sequence selected from SEQ ID NO 374, 375, 376, or 377. In certain such embodiments, a short antisense compound targeted to nucleotides 1621-1637 of SEQ ID NO: 4 is selected from ISIS NO 400342,400343,400344, or 400345.

In certain embodiments, a target region is nucleotides 1738-1754 of SEQ ID NO: 4. In certain such embodiments, a short antisense compound targeted to nucleotides 1738-1754 of SEQ ID NO: 4 comprises a nucleotide sequence selected from SEQ ID NO 378, 379, 380, or 381. In certain such embodiments, a short antisense compound targeted to nucleotides 1738-1754 of SEQ ID NO: 4 is selected from ISIS NO 400346,400347,400348, or 400349.

In certain embodiments, a target region is nucleotides 1834-1853 of SEQ ID NO: 4. In certain such embodiments, a short antisense compound targeted to nucleotides 1834-1853 of SEQ ID NO: 4 comprises a nucleotide sequence selected from SEQ ID NO 382, 383, 384, 385, 386, 387, or 388. In certain embodiments, a short antisense compound targeted to nucleotides 1834-1853 of SEQ ID NO: 4 is selected from ISIS NO 400350,400351,400352,400353,400354,400355, or 400356.

In certain embodiments, a target region is nucleotides 2083-2099 of SEQ ID NO: 4. In certain such embodiments, a short antisense compound targeted to nucleotides 2083-2099 of SEQ ID NO: 4 comprises a nucleotide sequence selected from SEQ ID NO 389, 390, 391, or 392. In certain such embodiments, a short antisense compound targeted to nucleotides 2083-2099 of SEQ ID NO: 4 is selected from ISIS NO 400357,400358,400359, or 400360.

In certain embodiments, a target region is nucleotides 2316-2338 of SEQ ID NO: 4. In certain such embodiments, a short antisense compound targeted to nucleotides 2316-2338 of SEQ ID NO: 4 comprises a nucleotide sequence selected from SEQ ID NO 393,394,395,396, 397,398, 399,400,401, or 402. In certain such embodiments, a short antisense compound targeted to nucleotides 2316-2338 of SEQ ID NO: 4 is selected from ISIS NO 400361, 400362, 400363, 400364, 400365, 400366, 400367, 400368,400369, or 400370.

In certain embodiments, short antisense compounds targeted to a PCSK9 nucleic acid are 8 to 16, preferably 9 to 15, more preferably 9 to 14, more preferably 10 to 14 nucleotides in length. In certain embodiments, short antisense compounds targeted to a PCSK9 nucleic acid are 9 to 14 nucleotides in length. In certain embodiments, short antisense compounds targeted to a PCSK9 nucleic acid are 10 to 14 nucleotides in length. In certain embodiments, such short antisense compounds are short antisense oligonucleotides.

In certain embodiments, short antisense compounds targeted to a PCSK9 nucleic acid are short gapmers. In certain such embodiments, short gapmers targeted to a PCSK9 nucleic acid comprise at least

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-88one high affinity modification in one or more wings of the compound. In certain embodiments, short antisense compounds targeted to a PCSK9 nucleic acid comprise 1 to 3 high-affinity modifications in each wing. In certain such embodiments, the nucleosides or nucleotides of the wing comprise a 2’ modification. In certain such embodiments, the monomers of the wing are BNA’s. In certain such embodiments, the monomers of the wing are selected from α-L-Methyleneoxy (4’-CH₂-O-2’) BNA, β-DMethyleneoxy (4’-CH_rO-2’) BNA , Ethyleneoxy (4’-(CH₂)₂-O-2’) BNA , Aminooxy (4’-CH₂-O-N(R)2’) BNA and Oxyamino (4’-CH₂-N(R)-O-2’) BNA. In certain embodiments, the monomers of a wing comprise a substituent at the 2’ position selected from allyl, amino, azido, thio, O-allyl, 0-C_rC|o alkyl, OCFj, O-(CH₂)₂-O-CH₃, Z'-OiC^hSCHj.O-iCH^-O-NiRnOiR,), and O-CH₂-C(=O)-N(R_M)(R_n), where each R_ro and R„ is, independently, H or substituted or unsubstituted Ci-Cio alkyl. In certain embodiments, the monomers of a wing are 2’MOE nucleotides.

In certain embodiments, short antisense compounds targeted to a PCSK9 nucleic acid comprise a gap between the 5’ wing and the 3’ wing. In certain embodiments the gap comprises five, six, seven, eight, nine, ten, eleven, twelve, thirteen, or fourteen monomers. In certain embodiments, the monomers of the gap are unmodified deoxyribonucleotides. In certain embodiments, the monomers of the gap are unmodified ribonucleotides. In certain embodiments, gap modifications (if any) gap result in an antisense compound that, when bound to its target nucleic acid, supports cleavage by an RNase, including, but not limited to, RNase H.

In certain embodiments, short antisense compounds targeting a PCSK.9 nucleic acid may have any one or more properties or characteristics of the short antisense compounds generally described herein. In certain embodiments, short antisense compounds targeting a PCSK9 nucleic acid have a motif (wing deoxy gap -wing) selected from 1-12-1,1-1-10-2,2-10-1-1,3-10-3,2-10-3,2-10-2,1-10-1,1-10-2, 3-83,2-8-2,1-8-1,3-6-3 or 1-6-1, more preferably 1-10-1,2-10-2,3-10-3, and 1-9-2.

In certain embodiments, short antisense compounds targeted to a PCSK9 nucleic acid have uniform monomeric linkages. In certain such embodiments, those linkages are all phosphorothioate linkages. In certain embodiments, the linkages are all phosphodiester linkages. In certain embodiments, short antisense compounds targeted to a PCSK9 nucleic acid have mixed backbones.

In certain embodiments, short antisense compounds targeted to a PCSK9 nucleic acid are 8 monomers in length. In certain embodiments, short antisense compounds targeted to a PCSK9 nucleic acid are 9 monomers in length. In certain embodiments, short antisense compounds targeted to a PCSK9 nucleic acid are 10 monomers in length. In certain embodiments, short antisense compounds targeted to a PCSK9 nucleic acid are 11 monomers in length. In certain embodiments, short antisense compounds targeted to a PCSK9 nucleic acid are monomers in length. In certain embodiments, short antisense compounds targeted to a PCSK9 nucleic acid are 13 monomers in length. In certain embodiments, short antisense compounds targeted to a PCSK9 nucleic acid are 14 monomers in length. In certain embodiments, short antisense compounds targeted to a PCSK9 nucleic acid are 15 monomers in length. In certain embodiments, short antisense compounds targeted to a PCSK9 nucleic acid are 16 monomers in

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-89lcngth. In certain embodiments, short antisense compounds targeted to a PCSK9 nucleic acid comprise 9 to 15 monomers. In certain embodiments, short antisense compounds targeted to a PCSK9 nucleic acid comprise 10 to 15 monomers. In certain embodiments, short antisense compounds targeted to a PCSK9 nucleic acid comprise 12 to 14 monomers. In certain embodiments, short antisense compounds targeted to a PCSK9 nucleic acid comprise 12 to 14 nucleotides a* nucleosides.

In certain embodiments, the invention provides methods of modulating expression of PCSK9. In certain embodiments, such methods comprise use of one or more short antisense compound targeted to a PCSK9 nucleic acid, wherein the short antisense compound targeted to a PCSK9 nucleic acid is from about 8 to about 16, preferably 9 to 15, more preferably 9 to 14, more preferably 10 to 14 monomers (i.e. from about 8 to about 16 linked monomers). One of ordinary skill in the art will appreciate that this comprehends methods of modulating expression of PCSK9 using one or more short antisense compounds targeted to a PCSK9 nucleic acid of 8,9,10,11,12,13,14,15 or 16 monomers.

In certain embodiments, methods of modulating PCSK9 comprise use of a short antisense compound targeted to a PCSK9 nucleic acid that is 8 monomers in length. In certain embodiments, methods of modulating PCSK9 comprise use of a short antisense compound targeted to a PCSK9 nucleic acid that is 9 monomers in length. In certain embodiments, methods of modulating PCSK9 comprise use of a short antisense compound targeted to a PCSK9 nucleic acid that is 10 monomers in length. In certain embodiments, methods of modulating PCSK9 comprise use of a short antisense compound targeted to a PCSK9 nucleic acid that is 11 monomers in length. In certain embodiments, methods of modulating PCSK9 comprise use of a short antisense compound targeted to a PCSK9 nucleic acid that is 12 monomers in length. In certain embodiments, methods of modulating PCSK9 comprise use of a short antisense compound targeted to a PCSK.9 nucleic acid that is 13 monomers in length. In certain embodiments, methods of modulating PCSK9 comprise use of a short antisense compound targeted to a PCSK9 nucleic acid that is 14 monomers in length. In certain embodiments, methods of modulating PCSK9 comprise use of a short antisense compound targeted to a PCSK9 nucleic acid that is 15 monomers in length. In certain embodiments, methods of modulating PCSK9 comprise use of a short antisense compound targeted to a PCSK9 nucleic acid that is 16 monomers in length.

In certain embodiments, methods of modulating expression of PCSK9 comprise use of a short antisense compound targeted to a PCSK9 nucleic acid comprising 9 to 15 monomers. In certain embodiments, methods of modulating expression of PCSK9 comprise use of a short antisense compound targeted to a PCSK.9 nucleic acid comprising 10 to 15 monomers. In certain embodiments, methods of modulating expression of PCSK9 comprise use of a short antisense compound targeted to a PCSK9 nucleic acid comprising 12 to 14 monomers. In certain embodiments, methods of modulating expression of PCSK9 comprise use of a short antisense compound targeted to a PCSK9 nucleic acid comprising 12 or 14 nucleotides or nucleosides.

4. Superoxide Dismutase 1 Enzyme fSODl)

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-90The enzymes known as the superoxide dismutases (SODs) provide defense against oxidative damage of biomolecules by catalyzing the dismutation of superoxide to hydrogen peroxide (H₂O₂) (Fridovich, Annu. Rev. Biochem., 1995, 64, 97-112). Two major classes of superoxide dismutases exist. One consists of a group of enzymes with active sites containing copper and zinc while the other class has either manganese or iron at the active site (Fridovich, Annu. Rev. Biochem., 1995,64,97-112).

Mutations in the superoxide dismutase 1 gene are associated with a dominantly-inherited form of amyotrophic lateral sclerosis (ALS, also known as Lou Gehrig’s disease) a disorder characterized by a selective degeneration of upper and lower motor neurons (Cleveland and Liu, Nat. Med., 2000, 6, 13201321). The deleterious effects of various mutations on superoxide dismutase 1 are most likely mediated through a gain of toxic function rather than a loss of superoxide dismutase 1 activity, as the complete absence of superoxide dismutase 1 in mice neither diminishes life nor provokes overt disease (Al-Chalabi and Leigh, Curr. Opin. Neurol., 2000,13, 397-405; Alisky and Davidson, Hum. Gene Ther., 2000, 11, 2315-2329).

Over 100 mutations of the human SOD1 gene have been identified, and altogether account for approximately 20% of familial amyotrophic lateral sclerosis (ALS) cases. Some mutations, such as the A4V mutation most commonly found in the United States, are highly lethal and result in survival only nine months from the onset of disease symptoms. Other mutations of SOD1 manifest in a slower disease course.

Definitions “SOD1” means the gene product or protein of which expression is to be modulated by administration of a short antisense compound.

“SOD1 nucleic acid” means any nucleic acid encoding SOD1. For example, in certain embodiments, a SOD1 nucleic acid includes, without limitations, a DNA sequence encoding SOD1, an RNA sequence transcribed from DNA encoding SOD1, and an mRNA sequence encoding SOD1.

“SOD1 mRNA” means an mRNA encoding SOD1.

SOD1 Therapeutic Indications

It has been discovered that antisense inhibition of superoxide dismutase 1 (SOD1) in an animal model of familial ALS reduces both SOD1 mRNA and protein, and further results in a slowing of disease progression and, importantly, increased survival time. Accordingly, in certain embodiments, the invention provides methods for the slowing of disease progression in an individual suffering from familial ALS by administering to such an individual a short antisense compound targeted to an SOD1 nucleic acid. In certain such embodiments, a short antisense compound targeted to SOD1 are delivered directly to the cerebrospinal fluid of the individual. In certain such embodiments, methods further comprise increasing survival time of an individual suffering from familial ALS. Slowing of disease progression is indicated by an improvement in one or more indicators of ALS disease progression, including, without limitation,

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-91the revised ALS functional rating scale, pulmonary function tests, and muscle strength measurements.

SOD1 Combination Therapies

In certain embodiments, one or more pharmaceutical compositions comprising a short antisense compound targeted to an SOD1 nucleic acid is co-administered with one or more other pharmaceutical agents. In certain embodiments, such one or more other pharmaceutical agents are designed to treat the same disease or condition as the one or more pharmaceutical compositions of the present invention. In certain embodiments, such one or more other pharmaceutical agents are designed to treat a different disease or condition as the one or more pharmaceutical compositions of the present invention. In certain embodiments, such one or more other pharmaceutical agents are designed to treat an undesired effect of one or more pharmaceutical compositions of the present invention. In certain embodiments, one or more pharmaceutical compositions of the present invention are co-administered with another pharmaceutical agent to treat an undesired effect of that other pharmaceutical agent. In certain embodiments, one or more pharmaceutical compositions of the present invention and one or more other pharmaceutical agents are administered at the same time. In certain embodiments, one or more pharmaceutical compositions of the present invention and one or more other pharmaceutical agents are administered at different times. In certain embodiments, one or more pharmaceutical compositions of the present invention and one or more other pharmaceutical agents are prepared together in a single formulation. In certain embodiments, one or more pharmaceutical compositions of the present invention and one or more other pharmaceutical agents are prepared separately.

In certain embodiments, a co-administered pharmaceutical agent is a nicotinic acid. In certain such embodiments, the nicotinic acid is selected from immediate release nicotinic acid, extended release nicotinic acid, and sustained release nicotinic acid.

In certain embodiments, a co-administered pharmaceutical agent is a fibric acid. In certain such embodiments, a fibric acid is selected from gemfibrozil, fenofibrate, clofibrate, bezafibrate, and ciprofibrate.

Further examples of pharmaceutical agents that may be co-administered with a pharmaceutical composition comprising a short antisense compound targeted to SOD1 include, but are not limited to, corticosteroids, including but not limited to prednisone; immunoglobulins, including, but not limited to intravenous immunoglobulin (IVIg); analgesics (e.g., acetaminophen); anti-inflammatory agents, including, but not limited to non-steroidal anti-inflammatory drugs (e.g., ibuprofen, COX-1 inhibitors, and COX-2, inhibitors); salicylates; antibiotics; antivirals; antifungal agents; antidiabetic agents (e.g., biguanides, glucosidase inhibitors, insulins, sulfonylureas, and thiazolidenediones); adrenergic modifiers; diuretics; hormones (e.g., anabolic steroids, androgen, estrogen, calcitonin, progestin, somatostan, and thyroid hormones); immunomodulators; muscle relaxants; antihistamines; osteoporosis agents (e.g., biphosphonates, calcitonin, and estrogens); prostaglandins, antineoplastic agents; psychotherapeutic agents; sedatives; poison oak or poison sumac products; antibodies; and vaccines.

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-92Certain Short Antisense Compounds Targeted to a SOD] Nucleic Acid

In certain embodiments, short antisense compounds are targeted to a SOD1 nucleic acid having the sequence of GENBANK® Accession No. NM_X02317.1, inccaporated herein as SEQ ID NO: 5. In certain such embodiments, a short antisense compound targeted to SEQ ID NO: 5 is at least 90% complementary to SEQ ID NO; 5. In certain such embodiments, a short antisense compound targeted to SEQ ID NO: 5 is at least 95% complementary to SEQ ID NO: 5. In certain such embodiments, a short antisense compound targeted to SEQ ID NO: 5 is 100% complementary to SEQ ID NO: 5. In certain embodiments, a short antisense compound targeted to SEQ ID NO: 5 comprises a nucleotide sequence selected from the nucleotide sequences set forth in Table 8 or Table 9.

The nucleotide sequence set forth in each SEQ ID NO in Tables 8 and 9 is independent of any modification to a sugar moiety, an intemucleoside linkage, or a nucleobase. As such, short antisense compounds defined by a SEQ ID NO may comprise, independently, one or more modifications to a sugar moiety, an intemucleoside linkage, or a nucleobase. Short antisense compounds described by Isis Number (Isis NO.) indicate a combination of nucleobase sequence and one or more modifications to a sugar moiety, an intemucleoside linkage, or a nucleobase.

Table 8 illustrates examples of short antisense compounds targeted to SEQ ID NO: 5. Table 8 illustrates short antisense compounds that are 100% complementary to SEQ ID NO: 5. The column labeled ‘gapmer motif indicates the wing-gap-wing motif of each short antisense compounds. The gap segment comprises 2’-deoxynucleotides and each nucleotide of each wing segment comprises a 2’modified sugar. The particular 2’-modified sugar is also indicated in the ‘gapmer motif column. For example, ‘2-10-2 MOE’ means a 2-10-2 gapmer motif, where a gap segment of ten 2’-deoxynucleotides is flanked by wing segments of two nucleotides, where the nucleotides of the wing segments are 2’-MOE nucleotides. Intemucleoside linkages are phosphorothioate. The short antisense compounds comprise 5methylcytidine in place of unmodified cytosine, unless “unmodified cytosine” is listed in the gapmer motif column, in which case the indicated cytosines are unmodified cytosines. For example, “5-mC in gap only” indicates that the gap segment has 5-mcthylcytosines, while the wing segments have unmodified cytosines.

In certain embodiments, short antisense compounds targeting a SOD1 nucleic acid may have any one or more properties or characteristics of the short antisense compounds generally described herein. In certain embodiments, short antisense compounds targeting a SOD1 nucleic acid have a motif (wing deoxy gap-wing) selected from 1-12-1,1-1-10-2,2-10-1-1,3-10-3,2-10-3,2-10-2,1-10-1,1-10-2, 3-83,2-8-2,1-8-1,3-6-3 or 1-6-1, more preferably 1-10-1,2-10-2,3-10-3, and 1-9-2.

Table 8: Short Antisense Compounds targeted to SEQ ID NO: 5

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<td> ISIS NO.</td><td> 5’ Target Site</td><td> 3’ Target Site</td><td> Sequence (5’-3’)</td><td> Gapmer Motif</td><td> SEQ ID NO</td>

<td> 387541</td><td> 85</td><td> 100</td><td> GTCGCCCTTCAGCACG</td><td> 3-10-3 MOE</td><td> 406</td>

<td> 387540</td><td> 86</td><td> 99</td><td> TCGCCCTTCAGCAC</td><td> 2-10-2 MOE</td><td> 407</td>

<td> 387539</td><td> 87</td><td> 98</td><td> CGCCCTTCAGCA</td><td> 1-10-1 MOE</td><td> 408</td>

In certain embodiments, a target region is nucleotides 85-100 of SEQ ID NO: 5. In certain such embodiments, short antisense compounds targeted to nucleotides 85-100 of SEQ ID NO: 5 comprise a nucleotide sequence selected from SEQ ID NO: 406,407, or 408. In certain such embodiments, a short antisense compound targeted to nucleotides 85-100 of SEQ ID NO: 5 is selected from Isis No. 387541, 387540, or 387539.

In certain embodiments, short antisense compounds targeted to a SOD1 nucleic acid are 8 to 16, preferably 9 to 15, more preferably 9 to 14, more preferably 10 to 14 nucleotides in length. In certain embodiments, short antisense compounds targeted to a SOD1 nucleic acid are 9 to 14 nucleotides in length. In certain embodiments, short antisense compounds targeted to a SOD1 nucleic acid are 10 to 14 nucleotides in length. In certain embodiments, such short antisense compounds are short antisense oligonucleotides.

In certain embodiments, short antisense compounds targeted to a SOD1 nucleic acid are short gapmers. In certain such embodiments, short gapmers targeted to a SOD1 nucleic acid comprise at least one high affinity modification in one or more wings of the compound. In certain embodiments, short antisense compounds targeted to a SOD1 nucleic acid comprise 1 to 3 high-affinity modifications in each wing. In certain such embodiments, the nucleosides or nucleotides of the wing comprise a 2’ modification. In certain such embodiments, the monomers of the wing are BNA’s. In certain such embodiments, the monomers of the wing are selected from a-L-Mcthylcncoxy (4’-CH₂-O-2’) BNA, β-DMethyleneoxy (4’-CH₂-O-2’) BNA , Ethyleneoxy (4’-(CH₂)2-O-2’) BNA , Aminooxy (4’-CH₂-O-N(R)2’) BNA and Oxyamino (4’-CH₂-N(R)-O-2’) BNA. In certain embodiments, the monomers of a wing comprise a substituent at the 2’ position selected from allyl, amino, azido, thio, O-allyl, 0-C_rCio alkyl, OCFj, O-(CH₂)rO-CH3, 2'-O(CH₂)₂SCH,, O-(CH₂)₂-O-N(R„)(R_n), and O-CH₂-C(=O)-N(RJ(R_n), where each R_m and Ιζ is, independently, H or substituted or unsubstituted C|-Cio alkyl. In certain embodiments, the monomers of a wing are 2’MOE nucleotides.

In certain embodiments, short antisense compounds targeted to a SOD1 nucleic acid comprise a gap between the 5’ wing and the 3’ wing. In certain embodiments the gap comprises five, six, seven, eight, nine, ten, eleven, twelve, thirteen, or fourteen monomers. In certain embodiments, the monomers of the gap are unmodified deoxyribonucleotides. In certain embodiments, the monomers of the gap are unmodified ribonucleotides. In certain embodiments, gap modifications (if any) gap result in an antisense compound that, when bound to its target nucleic acid, supports cleavage by an RNase, including, but not limited to, RNase H.

In certain embodiments, short antisense compounds targeted to a SOD1 nucleic acid have

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-94uniform monomeric linkages. In certain such embodiments, those linkages are all phosphorothioate linkages. In certain embodiments, the linkages are all phosphodiester linkages. In certain embodiments, short antisense compounds targeted to a SODI nucleic acid have mixed backbones.

In certain embodiments, short antisense compounds targeted to a SOD1 nucleic acid are 8 monomers in length. In certain embodiments, short antisense compounds targeted to a SODI nucleic acid are 9 monomers in length. In certain embodiments, short antisense compounds targeted to a SOD1 nucleic acid are 10 monomers in length. In certain embodiments, short antisense compounds targeted to a SOD1 nucleic acid are 11 monomers in length. In certain embodiments, short antisense compounds targeted to a SODI nucleic acid are monomers in length. In certain embodiments, short antisense compounds targeted to a SODI nucleic acid are 13 monomers in length. In certain embodiments, short antisense compounds targeted to a SODI nucleic acid are 14 monomers in length. In certain embodiments, short antisense compounds targeted to a SODI nucleic acid are 15 monomers in length. In certain embodiments, short antisense compounds targeted to a SODI nucleic acid are 16 monomers in length. In certain embodiments, short antisense compounds targeted to a SOD1 nucleic acid comprise 9 to 15 monomers. In certain embodiments, short antisense compounds targeted to a SOD1 nucleic acid comprise 10 to 15 monomers. In certain embodiments, short antisense compounds targeted to a SOD1 nucleic acid comprise 12 to 14 monomers. In certain embodiments, short antisense compounds targeted to a SOD1 nucleic acid comprise 12 to 14 nucleotides or nucleosides.

In certain embodiments, the invention provides methods of modulating expression of SODI. In certain embodiments, such methods comprise use of one or more short antisense compound targeted to a SODI nucleic acid, wherein the short antisense compound targeted to a SOD1 nucleic acid is from about 8 to about 16, preferably 9 to 15, more preferably 9 to 14, more preferably 10 to 14 monomers (i.e. from about 8 to about 16 linked monomers). One of ordinary skill in the art will appreciate that this comprehends methods of modulating expression of SOD1 using one or more short antisense compounds targeted toaSODl nucleic acid of 8,9,10,11,12,13,14,15 or 16 monomers.

In certain embodiments, methods of modulating SOD1 comprise use of a short antisense compound targeted to a SOD1 nucleic acid that is 8 monomers in length. In certain embodiments, methods of modulating SOD1 comprise use of a short antisense compound targeted to a SODI nucleic acid that is 9 monomers in length. In certain embodiments, methods of modulating SODI comprise use of a short antisense compound targeted to a SODI nucleic acid that is 10 monomers in length. In certain embodiments, methods of modulating SODI comprise use of a short antisense compound targeted to a SODI nucleic acid that is 11 monomers in length. In certain embodiments, methods of modulating SODI comprise use of a short antisense compound targeted to a SODI nucleic add that is 12 monomers in length. In certain embodiments, methods of modulating SODI comprise use of a short antisense compound targeted to a SODI nucleic acid that is 13 monomers in length. In certain embodiments, methods of modulating SODI comprise use of a short antisense compound targeted to a SODI nucleic acid that is 14 monomers in length. In certain embodiments, methods of modulating SODI comprise use

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-95of a short antisense compound targeted to a SOD1 nucleic acid that is 15 monomers in length. In certain embodiments, methods of modulating SOD1 comprise use of a short antisense compound targeted to a SOD1 nucleic acid that is 16 monomers in length.

In certain embodiments, methods of modulating expression of SOD1 comprise use of a short antisense compound targeted to a SOD1 nucleic acid comprising 9 to 15 monomers. In certain embodiments, methods of modulating expression of SOD1 comprise use of a short antisense compound targeted to a SOD1 nucleic acid comprising 10 to 15 monomers. In certain embodiments, methods of modulating expression of SOD1 comprise use of a short antisense compound targeted to a SOD1 nucleic acid comprising 12 to 14 monomers. In certain embodiments, methods of modulating expression of SOD1 comprise use of a short antisense compound targeted to a SOD1 nucleic acid comprising 12 or 14 nucleotides or nucleosides.

5. CRP

CRP (also known as C-reactive protein and PTX1) is an essential human acute-phase reactant produced in the liver in response to a variety of inflammatory cytokines. The protein, first identified in 1930, is highly conserved and considered to be an early indicator of infectious or inflammatory conditions. Plasma CRP levels increase 1,000-fold in response to infection, ischemia, trauma, bums, and inflammatory conditions. In clinical trials where patients receive lipid-lowering therapy, such as statin therapy, it has been demonstrated that patients having reductions in both LDL-C and CRP have a reduced risk of future coronary events relative to patients experiencing only reductions in LDVC.

Definitions “CRP” means the gene product or protein of which expression is to be modulated by a short antisense compound.

“CRP nucleic acid means any nucleic acid encoding CRP. For example, in certain embodiments, a CRP nucleic acid includes, without limitations, a DNA sequence encoding CRP, an RNA sequence transcribed from DNA encoding CRP, and an mRNA sequence encoding CRP.

“CRP mRNA” means an mRNA encoding CRP.

CRP Therapeutic Indications

In certain embodiments, the invention provides methods of modulating CRP expression in an individual comprising administering to the individual a short antisense compound targeted to a CRP nucleic acid. In certain embodiments, the invention provides methods of treating an individual comprising administering one or more pharmaceutical compositions comprising a short antisense compound targeted to a CRP nucleic acid. In certain embodiments, the individual has hypercholesterolemia, non-familial hypercholesterolemia, familial hypercholesterolemia, heterozygous familial hypercholesterolemia, homozygous familial hypercholesterolemia, mixed dyslipidemia, atherosclerosis, a risk of developing

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-96atherosclerosis, coronary heart disease, a history of coronary heart disease, early onset coronary heart disease, one or more risk factors for coronary heart disease. In certain embodiments, the individual has acute coronary syndrome, vascular injury, arterial occlusion, unstable angina, post peripheral vascular disease, post myocardial infarction (Ml), thrombosis, deep vein thrombus, end-stage renal disease (ESRD), chronic renal failure, complement activation, congestive heart failure, or systemic vasculitis. In certain embodiments, the individual has had a stroke.

In certain embodiments, the individual has undergone a procedure selected from elective stent placement, angioplasty, post percutaneous transluminal angioplasty (PTCA), cardiac transplantation, renal dialysis or cardiopulmonary bypass.

In certain embodiments, the individual has an inflammatory disease. In certain such embodiments, the inflammatory disease is selected from inflammatory bowel disease, ulcerative colitis, rheumatoid arthritis, or osteoarthritis.

Guidelines for lipid-lowering therapy were established in 2001 by Adult Treatment Panel III (ATP III) of the National Cholesterol Education Program (NCEP), and updated in 2004 (Gnindy et al., Circulation, 2004, 110, 227-239). The guidelines include obtaining a complete lipoprotein profile, typically after a 9 to 12 hour fast, for determination of LDL-C, total cholesterol, and HDL-C levels. According to the most recently established guidelines, LDL-C levels of 130-159 mg/dL, 160-189 mg/dL, and greater than or equal to 190 mg/dL are considered borderline high, high, and very high, respectively. Total cholesterol levels of 200-239 and greater than or equal to 240 mg/dL are considered borderline high and high, respectively. HDL-C levels of less than 40 mg/dL are considered low.

In certain embodiments, the individual has been identified as in need of lipid-lowering therapy. In certain such embodiments, the individual has been identified as in need of lipid-lowering therapy according to the guidelines established in 2001 by Adult Treatment Panel III (ATP III) of the National Cholesterol Education Program (NCEP), and updated in 2004 (Grundy et al., Circulation, 2004, 110,227239). In certain such embodiments, the individual in need of lipid-lowering therapy has LDI^C above 190 mg/dL. In certain such embodiments, the individual in need of lipid-lowering therapy has LDL-C above 160 mg/dL. In certain such embodiments, the individual in need of lipid-lowering therapy has LDL-C above 130 mg/dL. In certain such embodiments the individual in need of lipid-lowering therapy has LDLC above 100 mg/dL. In certain such embodiments the individual in need of lipid-lowering therapy should maintain LDL-C below 160 mg/dL. In certain such embodiments the individual in need of lipid-lowering therapy should maintain LDL-C below 130 mg/dL. In certain such embodiments the individual in need of lipid-lowering therapy should maintain LDIÆ below 100 mg/dL. In certain such embodiments the individual should maintain LDL-C below 70 mg/dL.

In certain embodiments the invention provides methods for reducing CRP in an individual. In certain such embodiments, the reduction in CRP is at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, and at least 100%.

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-97In certain embodiments, the methods provided by the present invention do not lower HDL-C. In certain embodiments, the methods provided by the present invention do not result in accumulation of lipids in the liver. In certain embodiments, the methods provided by the present invention do not cause hepatic steatosis.

In certain embodiments, the invention provides methods for lowering CRP concentration in a subject while reducing side effects associated with treatment. In certain such embodiments, a side effect is liver toxicity. In certain such embodiments, a side effect is abnormal liver function. In certain such embodiments, a side effect is elevated alanine aminotransferase (ALT). In certain such embodiments, a side effect is elevated aspartate aminotransferase (AST).

In certain embodiments, the invention provides methods for lowering CRP concentration in a subject who is not reaching target LDL-C levels as a result of lipid-lowering therapy. In certain such embodiments, a short antisense compound targeted to a CRP nucleic acid is the only pharmaceutical agent administered to the subject. In certain such embodiments, the subject has not complied with recommended lipid-lowering therapy. In certain such embodiments, a pharmaceutical composition of the invention is co-administered with an additional different lipid-lowering therapy. In certain such embodiments, an additional lipid-lowering therapy is LDL-apheresis. In certain such embodiments, an additional lipid-lowering therapy is a statin. In certain such embodiments, an additional lipid-lowering therapy is ezetimibe.

In certain embodiments, the invention provides methods for lowering CRP concentration in a statin-intolerant subject. In certain such embodiments, the subject has creatine kinase concentration increases as a result of statin administration. In certain such embodiments, the subject has liver function abnormalities as a result of statin administration. In certain such embodiments the subject has muscle aches as a result of statin administration. In certain such embodiments the subject has central nervous system side effects as a result of statin administration. In certain embodiments, the subject has not complied with recommended statin administration.

In certain embodiments, the invention provides methods for reducing coronary heart disease risk in a subject. In certain embodiments the invention provides methods for slowing the progression of atherosclerosis in a subject In certain such embodiments the invention provides methods for stopping the progression of atherosclerosis in a subject. In certain such embodiments the invention provides methods 30 for reducing the size and/or prevalence of atherosclerotic plaques in a subject. In certain embodiments the methods provided reduce a subject’s risk of developing atherosclerosis.

In certain embodiments the methods provided improve the cardiovascular outcome in a subject.

In certain such embodiments improved cardiovascular outcome is the reduction of the risk of developing coronary heart disease. In certain such embodiments, improved cardiovascular outcome is a reduction in 35 the occurance of one or more major cardiovascular events, which include, but are not limited to, death, myocardial infarction, reinfarction, stroke, cardiogenic shock, pulmonary edema, cardiac arrest, and atrial dysrhythmia. In certain such embodiments, the improved cardiovascular outcome is evidenced by

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-98improved carotid intimal media thickness. In certain such embodiments, improved carotid intimal media thickness is a decrease in thickness. In certain such embodiments, improved carotid intimal media thickness is a prevention an increase of intimal media thickness.

In certain embodiments a pharmaceutical composition comprising a short antisense compound targeted to a CRP nucleic acid is for use in therapy. In certain embodiments, the therapy is the reduction of CRP in an individual. In certain embodiments, the therapy is the treatment of hypercholesterolemia, non-familial hypercholesterolemia, familial hypercholesterolemia, heterozygous familial hypercholesterolemia, homozygous familial hypercholesterolemia, mixed dyslipidemia, atherosclerosis, a risk of developing atherosclerosis, coronary heart disease, a history of coronary heart disease, or early onset coronary heart disease. In additional embodiments, the therapy is the reduction of CHD risk. In certain the therapy is prevention of atherosclerosis. In certain embodiments, the therapy is the prevention of coronary heart disease. In certain embodiments, the therapy is the treatment of acute coronary syndrome, chronic renal failure, vascular injury, arterial occlusion, atherothrombosis, unstable angina, post peripheral vascular disease, post myocardial infarction (MI), thrombosis, deep vein thrombus, endstage renal disease (ESRD), complement activation, congestive heart failure, or systemic vasculitis. In certain embodiments the therapy is the treatment of an individual who has undergone a procedure selected from elective stent placement, angioplasty, post percutaneous transluminal angioplasty (PTCA), cardiac transplantation, renal dialysis or cardiopulmonary bypass. In certain embodiments, the therapy is the treatment of an inflammatory disorder.

In certain embodiments a pharmaceutical composition comprising a short antisense compound targeted to a CRP nucleic acid is used for the preparation of a medicament for reducing CRP in an individual. In certain embodiments pharmaceutical composition comprising a short antisense compound targeted to a CRP nucleic acid is used for the preparation of a medicament for reducing coronary heart disease risk. In certain embodiments a short antisense compound targeted to a CRP nucleic acid is used for the preparation of a medicament for the treatment of hypercholesterolemia, non-familial hypercholesterolemia, familial hypercholesterolemia, heterozygous familial hypercholesterolemia, homozygous familial hypercholesterolemia, mixed dyslipidemia, atherosclerosis, a risk of developing atherosclerosis, coronary heart disease, a history of coronary heart disease, early onset coronary heart disease, or one or more risk factors for coronary heart disease.

In certain embodiments, a short antisense compound targeted to a CRP nucleic acid is used for the preparation of a medicament for die treatment of acute coronary syndrome, chronic renal failure, vascular injury, arterial occlusion, atherothrombosis, unstable angina, post peripheral vascular disease, post myocardial infarction (MI), thrombosis, deep vein thrombus, end-stage renal disease (ESRD), complement activation, congestive heart failure, or systemic vasculitis. In certain embodiments, a short antisense compound targeted to a CRP nucleic acid is used for the preparation of a medicament for the treatment of an individual who has had a stroke.

In certain embodiments, a short antisense compound targeted to a CRP nucleic acid is used for

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-99the preparation of a medicament for the treatment in an individual who has undergone a procedure selected from elective stent placemen angioplasty, post percutaneous transluminal angioplasty (PTCA), cardiac transplantation, renal dialysis or cardiopulmonary bypass.

In certain embodiments, a short antisense compound targeted to a CRP nucleic acid is used for the preparation of a medicament for the treatment of an inflammatory disease. In certain such embodiments, a short antisense compound targeted to a CRP nucleic acid is used for the preparation of a medicament for the treatment of inflammatory bowel disease, ulcerative colitis, rheumatoid arthritis, or osteoarthritis.

CRP Combination Therapies

In certain embodiments, one or more pharmaceutical compositions comprising a short antisense compound targeted to a CRP nucleic acid are co-administered with one or more other pharmaceutical agents. In certain embodiments, the one or more other pharmaceutical agents is a lipid-lowering agent. In certain embodiments, such one or more other pharmaceutical agents are designed to treat the same disease or condition as the one or more pharmaceutical compositions of the present invention. In certain embodiments, such one or more other pharmaceutical agents are designed to treat a different disease or condition as the one or more pharmaceutical compositions of the present invention. In certain embodiments, such one or more other pharmaceutical agents are designed to treat an undesired effect of one or more pharmaceutical compositions of the present invention. In certain embodiments, one or more pharmaceutical compositions of the present invention are co-administered with another pharmaceutical agent to treat an undesired effect of that other pharmaceutical agent. In certain embodiments, one or more pharmaceutical compositions of the present invention and one or more other pharmaceutical agents are administered at the same time. In certain embodiments, one or more pharmaceutical compositions of the present invention and one or more other pharmaceutical agents are administered at different times. In certain embodiments, one or more pharmaceutical compositions of the present invention and one or more other pharmaceutical agents are prepared together in a single formulation. In certain embodiments, one or more pharmaceutical compositions of the present invention and one or more other pharmaceutical agents are prepared separately.

In certain embodiments, pharmaceutical agents that may be co-administered with a pharmaceutical composition comprising a short antisense compound targeted to a CRP nucleic acid include lipid-lowering agents. In certain such embodiments, pharmaceutical agents that may be coadministered with a pharmaceutical composition of the present invention include, but are not limited to atorvastatin, simvastatin, rosuvastatin, and ezetimibe. In certain such embodiments, the lipid-lowering agent is administered prior to administration of a pharmaceutical composition of the present invention. In certain such embodiments, the lipid-lowering agent is administered following administration of a pharmaceutical composition of the present invention. In certain such embodiments the lipid-lowering agent is administered at the same time as a pharmaceutical composition of the present invention. In

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-100certain such embodiments the dose of a co-administered lipid-lowcring agent is the same as the dose that would be administered if the lipid-lowering agent was administered alone. In certain such embodiments the dose of a co-administered lipid-lowering agent is lower than the dose that would be administered if the lipid-lowering agent was administered alone. In certain such embodiments the dose of a coadministered lipid-lowering agent is greater than the dose that would be administered if the lipid-lowering agent was administered alone.

In certain embodiments, a co-administered lipid-lowcring agent is a HMG-CoA reductase inhibitor. In certain such embodiments the HMG-CoA reductase inhibitor is a statin. In certain such embodiments the statin is selected from atorvastatin, simvastatin, pravastatin, fluvastatin, and rosuvastatin.

In certain embodiments, a co-administered lipid-lowering agent is ISIS 301012.

In certain embodiments, a co-administered lipid-lowering agent is a cholesterol absorption inhibitor. In certain such embodiments, cholesterol absorption inhibitor is ezetimibe.

In certain embodiments, a co-administered lipid-lowering agent is a co-formulated HMG-CoA reductase inhibitor and cholesterol absorption inhibitor. In certain such embodiments the co-formulated lipid-lowering agent is ezetimibe/simvastatin.

In certain embodiments, a co-administered lipid-lowering agent is a microsomal triglyceride transfer protein inhibitor (MTP inhibitor).

In certain embodiments, a co-administered pharmaceutical agent is a bile acid séquestrant In certain such embodiments, the bile acid séquestrant is selected from cholestyramine, colestipol, and colesevelam.

In certain embodiments, a co-administered pharmaceutical agent is a nicotinic acid. In certain such embodiments, the nicotinic acid is selected from immediate release nicotinic acid, extended release nicotinic acid, and sustained release nicotinic acid.

In certain embodiments, a co-administered pharmaceutical agent is a fibric acid. In certain such embodiments, a fibric acid is selected from gemfibrozil, fenofibrate, clofibrate, bezafibrate, and ciprofibratc.

Further examples of pharmaceutical agents that may be co-administered with a pharmaceutical composition comprising a short antisense compound targeted to a CRP nucleic acid include, but are not limited to, corticosteroids, including but not limited to prednisone; immunoglobulins, including, but not limited to intravenous immunoglobulin (IVIg); analgesics (e.g., acetaminophen); anti-inflammatory agents, including, but not limited to non-steroidal anti-inflammatory drugs (e.g., ibuprofen, COX-1 inhibitors, and COX-2, inhibitors); salicylates; antibiotics; antivirals; antifungal agents; antidiabetic agents (e.g., biguanides, glucosidase inhibitors, insulins, sulfonylureas, and thiazolidenediones); adrenergic modifiers; diuretics; hormones (e.g., anabolic steroids, androgen, estrogen, calcitonin, progestin, somatostan, and thyroid hormones); immunomodulators; muscle relaxants; antihistamines; osteoporosis agents (e.g., biphosphonates, calcitonin, and estrogens); prostaglandins, antineoplastic

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-101agents; psychotherapeutic agents; sedatives; poison oak or poison sumac products; antibodies; and vaccines.

In certain embodiments, a pharmaceutical composition comprising a short antisense compound targeted to a CRP nucleic acid may be administered in conjuction with a lipid-lowering therapy. In certain such embodiments, a lipid-lowering therapy is therapeutic lifestyle change. In certain such embodiments, a lipid-lowering therapy is LDL apheresis.

Certain Short Antisense Compounds Targeted to a CRP nucleic acid

In certain embodiments, short antisense compounds are targeted to a CRP nucleic acid having the sequence of GENBANK® Accession No. NM_000567.1, incorporated herein as SEQ ID NO: 6. In certain such embodiments, a short antisense compound targeted to SEQ ID NO: 6 is at least 90% complementary to SEQ ID NO: 6. In certain such embodiments, a short antisense compound targeted to SEQ ID NO: 6 is at least 95% complementary to SEQ ID NO: 6. In certain such embodiments, a short antisense compound targeted to SEQ ID NO: 6 is 100% complementary to SEQ ID NO: 6. In certain embodiments, a short antisense compound targeted to SEQ ID NO: 6 comprises a nucleotide sequence selected from the nucleotide sequences set forth in Table 9.

The nucleotide sequence set forth in each SEQ ID NO in Table 9 is independent of any modification to a sugar moiety, an intemucleoside linkage, or a nucleobase. As such, short antisense compounds defined by a SEQ ID NO may comprise, independently, one or more modifications to a sugar moiety, an intemucleoside linkage, or a nucleobase. Short antisense compounds described by Isis Number (Isis NO.) indicate a combination of nucleobase sequence and one or more modifications to a sugar moiety, an intemucleoside linkage, or a nucleobase.

Table 9 illustrates examples of short antisense compounds targeted to SEQ ID NO: 6. Table 9 illustrates short antisense compounds that are 100% complementary to SEQ ID NO: 6. The column labeled ‘gapmer motif indicates the wing-gap-wing motif of each short antisense compounds. The gap segment comprises 2’-deoxynucleotides and each nucleotide of each wing segment comprises a 2’modified sugar. The particular 2’-modified sugar is also indicated in the ‘gapmer motif column. For example, ‘2-10-2 MOE’ means a 2-10-2 gapmer motif, where a gap segment of ten 2’-deoxynucleotides is flanked by wing segments of two nucleotides, where the nucleotides of the wing segments are 2’-MOE nucleotides. Intemucleoside linkages are phosphorothioate. The short antisense compounds comprise 5methylcytidine in place of unmodified cytosine, unless “unmodified cytosine” is listed in the gapmer motif column, in which case the indicated cytosines are unmodified cytosines. For example, “5-mC in gap only” indicates that the gap segment has 5-methylcytosines, while the wing segments have unmodified cytosines.

In certain embodiments, short antisense compounds targeting a CRP nucleic acid may have any one or more properties or characteristics of the short antisense compounds generally described herein. In certain embodiments, short antisense compounds targeting a CRP nucleic acid have a motif (wing CA 02651309 2008-11-04

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-102deoxy gap-wing) selected from 1-12-1,1-1-10-2,2-10-1-1,3-10-3,2-10-3,2-10-2,1-10-1,1-10-2, 3-83,2-8-2,1-8-1, 3-6-3 or 1-6-1, more preferably 1-10-1,2-10-2, 3-10-3, and 1-9-2.

Table 9: Short Antisense Compounds targeted to SEQ Π) NO: 6

<td> ISIS NO.</td><td> 5’ Target Site</td><td> 3' Target Site</td><td> Sequence (5’-3’)</td><td> Gapmer Motif</td><td> Seq Π) NO</td>

<td> 353506</td><td> 1257</td><td> 1272</td><td> ACTCTGGACCCAAACC</td><td> 3-10-3 MOE</td><td> 409</td>

<td> 353507</td><td> 1258</td><td> 1271</td><td> CTCTGGACCCAAAC</td><td> 2-10-2 MOE</td><td> 410</td>

<td> 353484</td><td> 1305</td><td> 1320</td><td> CCATTTCAGGAGACCT</td><td> 3-10-3 MOE</td><td> 411</td>

<td> 353485</td><td> 1306</td><td> 1319</td><td> CATTTCAGGAGACC</td><td> 2-10-2 MOE</td><td> 412</td>

In certain embodiments, a target region is nucleotides 1305-1320 of NM 000567.1. In certain such embodiments, short antisense compounds targeted to nucleotides 1305-1320 of NM_000567 1 comprise a nucleotide sequence selected from SEQ ID NO: 1305 or 1306. In certain such embodiments, a short antisense compound targeted to nucleotides 263-278 of NM_000567.1 is selected from Isis NO. 353484 or 353485.

In certain embodiments, a target region is nucleotides 1257-1272 of NM_000567.1. In certain such embodiments, a short antisense compound targeted to nucleotides 1257-1272 of NM_000567.1 comprises a nucleotide sequence selected from SEQ ID NO 1257 or 1258. In certain such embodiments, a short antisense compound targeted to nucleotides 428-483 of NM_000567.l is selected from Isis NO. 353506 or 353507.

In certain embodiments, short antisense compounds targeted to a CRP nucleic acid are 8 to 16, preferably 9 to 15, more preferably 9 to 14, more preferably 10 to 14 nucleotides in length. In certain embodiments, short antisense compounds targeted to a CRP nucleic acid are 9 to 14 nucleotides in length. In certain embodiments, short antisense compounds targeted to a CRP nucleic acid are 10 to 14 nucleotides in length. In certain embodiments, such short antisense compounds are short antisense oligonucleotides.

In certain embodiments, short antisense compounds targeted to a CRP nucleic acid are short gapmers. In certain such embodiments, short gapmers targeted to a CRP nucleic acid comprise at least one high affinity modification in one or more wings of the compound. In certain embodiments, short antisense compounds targeted to a CRP nucleic acid comprise 1 to 3 high-affinity modifications in each wing. In certain such embodiments, the nucleosides or nucleotides of the wing comprise a 2’ modification. In certain such embodiments, the monomers of the wing are BNA’s. In certain such embodiments, the monomers of the wing are selected from α-L-Methyleneoxy (4’-CH₂-O-2’) BNA, β-DMethyleneoxy (4’-CH₂-O-2’) BNA , Ethyleneoxy (4’-(CH_?),-O-2’) BNA , Aminooxy (4’-CH₂-O-N(R)2’) BNA and Oxyamino (4’-CH₂-N(R)-O-2’) BNA. In certain embodiments, the monomers of a wing comprise a substituent at the 2’ position selected from allyl, amino, azido, thio, O-allyl, O-Ci-Cio alkyl, OCFj, O-(CH₂)₂-O-CH₃, 2’-O(CH₂)2SCH3,O-(CH₂)₂-O-N(R_niXR_n), and O-C^-C^OXNiR»)^), where

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-103each Rm and R, is, independently, H or substituted or unsubstituted Ci-Cio alkyl. In certain embodiments, the monomers of a wing are 2’MOE nucleotides.

In certain embodiments, short antisense compounds targeted to a CRP nucleic acid comprise a gap between the 5’ wing and the 3’ wing. In certain embodiments the gap comprises five, six, seven, eight, nine, ten, eleven, twelve, thirteen, or fourteen monomers. In certain embodiments, the monomers of the gap are unmodified deoxyribonucleotides. In certain embodiments, the monomers of the gap are unmodified ribonucleotides. In certain embodiments, gap modifications (if any) gap result in an antisense compound that, when bound to its target nucleic acid, supports cleavage by an RNase, including, but not limited to, RNase H.

In certain embodiments, short antisense compounds targeted to a CRP nucleic acid have uniform monomeric linkages. In certain such embodiments, those linkages are all phosphorothioate linkages. In certain embodiments, the linkages are all phosphodiester linkages. In certain embodiments, short antisense compounds targeted to a CRP nucleic acid have mixed backbones.

In certain embodiments, short antisense compounds targeted to a CRP nucleic acid are 8 monomers in length. In certain embodiments, short antisense compounds targeted to a CRP nucleic acid are 9 monomers in length. In certain embodiments, short antisense compounds targeted to a CRP nucleic acid are 10 monomers in length. In certain embodiments, short antisense compounds targeted to a CRP nucleic acid are 11 monomers in length. In certain embodiments, short antisense compounds targeted to a CRP nucleic acid are monomers in length. In certain embodiments, short antisense compounds targeted to a CRP nucleic acid are 13 monomers in length. In certain embodiments, short antisense compounds targeted to a CRP nucleic acid are 14 monomers in length. In certain embodiments, short antisense compounds targeted to a CRP nucleic acid are 15 monomers in length. In certain embodiments, short antisense compounds targeted to a CRP nucleic acid are 16 monomers in length. In certain embodiments, short antisense compounds targeted to a CRP nucleic acid comprise 9 to 15 monomers. In certain embodiments, short antisense compounds targeted to a CRP nucleic acid comprise 10 to 15 monomers. In certain embodiments, short antisense compounds targeted to a CRP nucleic acid comprise 12 to 14 monomers. In certain embodiments, short antisense compounds targeted to a CRP nucleic acid comprise 12 to 14 nucleotides or nucleosides.

In certain embodiments, the invention provides methods of modulating expression of CRP. In certain embodiments, such methods comprise use of one or more short antisense compound targeted to a CRP nucleic acid, wherein the short antisense compound targeted to a CRP nucleic acid is from about 8 to about 16, preferably 9 to 15, more preferably 9 to 14, more preferably 10 to 14 monomers (i.e. from about 8 to about 16 linked monomers). One of ordinary skill in the art will appreciate that this comprehends methods of modulating expression of CRP using one or more short antisense compounds targeted to a CRP nucleic acid of 8,9,10,11,12,13,14,15 or 16 monomers.

In certain embodiments, methods of modulating CRP comprise use of a short antisense compound targeted to a CRP nucleic acid that is 8 monomers in length. In certain embodiments, methods

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-104of modulating CRP comprise use of a short antisense compound targeted to a CRP nucleic acid that is 9 monomers in length. In certain embodiments, methods of modulating CRP comprise use of a short antisense compound targeted to a CRP nucleic acid that is 10 monomers in length. In certain embodiments, methods of modulating CRP comprise use of a short antisense compound targeted to a CRP nucleic acid that is 11 monomers in length. In certain embodiments, methods of modulating CRP comprise use of a short antisense compound targeted to a CRP nucleic acid that is 12 monomers in length. In certain embodiments, methods of modulating CRP comprise use of a short antisense compound targeted to a CRP nucleic acid that is 13 monomers in length. In certain embodiments, methods of modulating CRP comprise use of a short antisense compound targeted to a CRP nucleic acid that is 14 monomers in length. In certain embodiments, methods of modulating CRP comprise use of a short antisense compound targeted to a CRP nucleic acid that is 15 monomers in length. In certain embodiments, methods of modulating CRP comprise use of a short antisense compound targeted to a CRP nucleic acid that is 16 monomers in length.

In certain embodiments, methods of modulating expression of CRP comprise use of a short antisense compound targeted to a CRP nucleic acid comprising 9 to 15 monomers. In certain embodiments, methods of modulating expression of CRP comprise use of a short antisense compound targeted to a CRP nucleic acid comprising 10 to 15 monomers. In certain embodiments, methods of modulating expression of CRP comprise use of a short antisense compound targeted to a CRP nucleic acid comprising 12 to 14 monomers. In certain embodiments, methods of modulating expression of CRP comprise use of a short antisense compound targeted to a CRP nucleic acid comprising 12 or 14 nucleotides or nucleosides.

6, Glucocorticoid Receptor (GCCR)

Glucocorticoids were among the first steroid hormones to be identified and are responsible for a multitude of physiological functions, including the stimulation of gluconeogenesis, decreased glucose uptake and utilization in peripheral tissues, increased glycogen deposition, suppression of immune and inflammatory responses, inhibition of cytokine synthesis and acceleration of various developmental events. Glucocorticoids are also especially important for combating stress. Stress-induced elevation of glucocorticoid synthesis and release leads to, among other responses, increased ventricular workload, inhibition of inflammatory mediators, inhibition of cytokine synthesis and increased glucose production (Karin, Cell, 1998,93,487-490).

Both natural glucocorticoids and their synthetic derivatives exert their action through the glucocorticoid receptor, a ubiquitously expressed cytoplasmic member of the nuclear hormone superfamily of receptors. Human glucocorticoid receptor is also known as nuclear receptor subfamily 3, group C, member 1; NR3C1; GCCR; GCR; GRL; Glucocorticoid receptor, lymphocyte. The gene is located on human chromosome 5ql l-ql3 and consists of 9 exons (Encio and Detera-Wadleigh, J Biol Chem, 1991,266,7182-7188; Gehring et al., Proc Natl Acad Sci USA, 1985,82, 3751-3755). Multiple

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-105forms of human glucocorticoid receptor mRNA exist: a 5.5 kb human glucocorticoid receptor a cDNA containing exons 1-8 and exon 9a; a 4.3 kb human glucocorticoid receptor β cDNA containing exons 1-8 and exon 9β; and a 7.0 kb human glucocorticoid receptor a cDNA containing exons 1-8 and the entire exon 9, which includes exon 9a, exon 9β and the ‘J region’, which is flanked by exons 9a and 9β (Hollenberg et al., Nature, 1985, 318, 635-641; Oakley et al., J Biol Chem, 1996, 271, 9550-9559). Human glucocorticoid receptor a is the predominant isoform of the receptor and the one that exhibits steroid binding activity (Hollenberg et al., Nature, 1985, 318, 635-641). Additionally, through usage of three different promoters three different exon 1 variants can be transcribed, and alternative splicing of one exon 1 variant can result in three different versions of this exon. Thus, human glucocorticoid receptor mRNA may contain 5 different versions of exon 1 (Breslin et al., Mol Endocrinol, 2001,75,1381-1395).

Examination of the expression patterns of the a and β isoforms of human glucocorticoid receptor mRNA reveals that the a isoform is more abundantly expressed. Both isoforms are expressed in similar tissues and cell types, including lung, kidney, heart, liver, skeletal muscle, macrophages, neutrophils and peripheral blood mononuclear cells. Only human glucocorticoid receptor a is expressed in colon. At the level of protein, while the a isoform is detected in all tissues examined, the β isoform is undetectable, suggesting that under physiological conditions, the default splicing pathway is the one that produces the a isoform (Pujols et al., Am J Physiol Cell Physiol, 2002, 283, C1324-1331). The β isoform of glucocorticoid receptor binds neither a glucocorticoid agonist nor an antagonist. Furthermore, the β isoform is localized primarily in the nucleus in transfected cells, independent of hormone stimulation. When both isoforms are expressed in the same cell, the glucocorticoid receptor β inhibits the hormoneinduced, glucocorticoid receptor α-mediated stimulation of gene expression, suggesting that the β isoform functions as an inhibitor of glucocorticoid receptor a activity (Oakley et ai, J Biol Chem, 1996, 271, 9550-9559). Unless otherwise noted, the human glucocorticoid receptor described herein is defined as the ubiquitous product(s) of the gene located on chromosome 5ql l-ql3.

Cell lines transfected with a complementary glucocorticoid receptor antisense RNA strand exhibited a reduction in glucocorticoid receptor mRNA levels and a decreased response to the glucocorticoid receptor agonist dexamethasone (Pepin and Barden, Mol Cell Biol, 1991, 77, 1647-1653). Transgenic mice bearing an antisense glucocorticoid receptor gene construct were used to study the glucocorticoid feedback effect on the hypothalamus-pituitary-adrenal axis (Pepin et al., Nature, 1992, 355,725-728). In another study of similarly genetically engineered mice, energy intake and expenditure, heart and vastus lateralis muscle lipoprotein lipase activity, and heart and brown adipose tissue norepinephrine were lower than in control animals. Conversely, fat content and total body energy were significantly higher than in control animals. These results suggest that a defective glucocorticoid receptor system may affect energy balance through increasing energetic efficiency, and they emphasize the modulatory effects of hypothalamic-pituitary-adrenal axis changes on muscle lipoprotein lipase activity (Richard et al., Am J Physiol, 1993,265, R146-150).

Behavorial effects of glucocorticoid receptor antagonists have been measured in animal models

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-106designed to assess anxiety, learning and memory. Reduced expression of glucocorticoid receptor in rats long-term intracerebroventricularly infused with antisense oligodeoxynucleotides targeting glucocorticoid receptor mRNA did not interfere with spatial navigation in the Morris water maze test (Engelmann et al., Eur J Pharmacol, 1998, 361, 17-26). Bilateral infusion of an antisense oligodeoxynucleotide targeting the glucocorticoid receptor mRNA into the dentate gyrus of the rat hippocampus reduced the immobility of rats in the Porsolt forced swim test (Korte et al., Eur J Pharmacol, 1996,301,19-25).

Glucocorticoids are frequently used for their immunosuppressive, anti-inflammatory effects in the treatment of diseases such as allergies, athsma, rheumatoid arthritis, AIDS, systemic lupus erythematosus and degenerative osteoarthritis. Negative regulation of gene expression, such as that caused by the interaction of glucocorticoid receptor with NF-kB, is proposed to be at least partly responsible for the anti-inflammatory action of glucocorticoids in vivo. Interleukin-6, tumor necrosis factor a and interleukin-1 are the three cytokines that account for most of the hypothalamic-pituitary-adrenal (HPA) axis stimulation during the stress of inflammation. The HPA axis and the systemic sympathetic and adrenomedullary system are the peripheral components of the stress system, responsible for maintaining basal and strcss-related homeostasis. Glucocorticoids, the end products of the HPA axis, inhibit the production of all three inflammatory cytokines and also inhibit their effects on target tissues, with the exception of interleukin-6, which acts synergistically with glucocorticoids to stimulate the production of acute-phase reactants. Glucocorticoid treatment decreases the activity of the HPA axis (Chrousos, N Engl J Med, 1995,332, 1351-1362).

In some cases, patients are refractory to glucocorticoid treatment One reason for this resistance to steroids lies with mutations or polymorphisms present in the glucocorticoid receptor gene. A total of 15 missense, three nonsense, three frameshift, one splice site, and two alternative spliced mutations, as well as 16 polymorphisms, have been reported in the NR3C1 gene in association with glucocorticoid resistance (Bray and Cotton, Hum Mutât, 2003, 21, 557-568). Additional studies in humans have suggested a positive association between metabolic syndrome incidence and progression, with alleles at the glucocorticoid receptor (GR) gene (Rosmond, Obes Res, 2002,10, 1078-1086).

Other cases of glucocorticoid insensitivity are associated with altered expression of glucocorticoid receptor isoforms. A study of human glucocorticoid receptor β isoform mRNA expression in glucocorticoid-resistant ulcerative colitis patients revealed the presence of this mRNA was significantly higher than in the glucocorticoid-sensitive patients, suggesting that the expression of human glucocorticoid receptor β mRNA in the peripheral blood mononuclear cells may serve as a predictor of glucocorticoid response in ulcerative colitis (Honda et al., Gastroenterology, 2000, 118, 859-866). Increased expression of glucocorticoid receptor β is also observed in a significantly high number of glucocorticoid-insensitive asthmatics. Additionally, cytokine-induced abnormalities in the DNA binding capacity of the glucocorticoid receptor were found in peripheral blood mononuclear cells from glucocorticoid-insensitive patients transfection, and HepG2 cells with the glucocorticoid receptor β gene resulted in a significant reduction of glucocorticoid receptor a DNA-binding capacity (Leung et al., J Exp

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-107Med, 1997, 186, 1567-1574). Dexamethasone binding studies demonstrate that human glucocorticoid receptor β does not alter the affinity of glucocorticoid receptor a for hormonal ligands, but rather its ability to bind to the GRE (Bamberger et al., J Clin Invest, 1995, 95, 2435-2441). Taken together, these results illustrate that glucocorticoid receptor β, through competition with glucocorticoid receptor a for GRE target sites, may function as a physiologically and pathophysiologically relevant endogenous inhibitor of glucocorticoid action.

In the liver, glucocorticoid agonists increase hepatic glucose production by activating the glucocorticoid receptor, which subsequently leads to increased expression of the gluconeogenic enzymes phosphoenolpyruvate carboxykinase (PEPCK) and glucose-6-phosphatase. Through gluconeogenesis, glucose is formed through non-hexose precursors, such as lactate, pyruvate and alanine (Link, Cun Opin Investig Drugs, 2003, 4, 421-429). Steroidal glucocorticoid receptor antagonists such as RU 486 have been tested in rodent models of diabetes. Mice deficient in the leptin receptor gene, termed db/db mice, are genetically obese, diabetic and hyperinsulinémie. Treatment of hyperglycemic db/db mice with RU 486 decreased blood glucose levels by approximately 49%, without affecting plasma insulin levels. Additionally, RU 486 treatment reduced the expression of glucocorticoid receptor responsive genes PEPCK, glucose-6-phosphatase, glucose transporter type 2 and tyrosine aminotransferase in db/db mice as compared to untreated animals (Friedman et al., J Biol Chem, 1997,272,31475-31481). RU 486 also ameliorates diabetes in the ob/ob mouse model of diabetes, obesity and hyperinsulinemia, through a reduction in serum insulin and blood glucose levels (Gettys et al., Int J Obes Relat Metab Disord, 1997, 21,865-873).

As increased gluconeogenesis is considered to be the major source of increased glucose production in diabetes, a number of therapeutic targets for the inhibition of hepatic glucose production have been investigated. Due to the ability of antagonists of the glucocorticoid receptor to ameliorate diabetes in animal models, such compounds are among the potential therapies being explored. However, there are detrimental systemic effects of glucocorticoid receptor antagonists, including activation of the HPA axis (Link, Curr Opin Investig Drugs, 2003,4,421-429). Increased HPA axis activity is associated with suppression of immune-related inflammatory action, which can increase susceptibility to infectious agents and neoplasms. Conditions associated with suppression of immune-mediated inflammation through defects in the HPA axis, or its target tissues, include Cushing’s syndrome, chronic stress, chronic alcoholism and melancholic depression (Chrousos, N Engl J Med, 1995, 332, 1351-1362). Thus, it is of great value to develop liver-specific glucocorticoid receptor antagonists. Steroidal glucocorticoid receptor antagonists have been conjugated to bile acids for the purpose of targeting them to the liver (Apelqvist et al., 2000). Currently, there are no known therapeutic agents that target the glucocorticoid receptor without undesired peripheral effects (Link, Curr Opin Investig Drugs, 2003, 4, 421-429). Consequently, there remains a long felt need for agents capable of effectively inhibiting hepatic glucocorticoid receptor.

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-108Definitions “Glucocorticoid receptor” is the gene product or protein of which expression is to be modulated by administration of a short antisense compound. Glucocorticoid receptor is generally referred to as GCCR.

“GCCR nucleic acid means any nucleic acid encoding GCCR. For example, in certain embodiments, a GCCR nucleic acid includes, without limitation, a DNA sequence encoding GCCR, an RNA sequence transcribed from DNA encoding GCCR, and an mRNA sequence encoding GCCR. “GCCR mRNA” means an mRNA encoding GCCR.

Therapeutic indications

Antisense technology is an effective means of reducing the expression of specific gene products and therefore is useful in a number of therapeutic, diagnostic and research applications for the modulation of glucocorticoid receptor expression. Furthermore, in certain embodiments, liver is one of the tissues in which the highest concentrations of antisense oligonucleotides are found following administration (Geary et al., Curr. Opin. Investig. Drugs, 2001, 2, 562-573). Therefore, in such embodiments, antisense technology represents an attractive method for the liver-specific inhibition of glucocorticoid receptor.

In certain embodiments, short antisense compounds targeted to a nucleic acid encoding glucocorticoid receptor are preferentially distributed to the liver. In certain embodiments, short antisense compounds have increased potency in the liver when compared to a longer parent compound. In certain embodiments, target RNA is predominantly expressed in the liver.

For therapeutics, a subject, suspected of having a disease or disorder which can be treated by modulating the expression of GCCR is treated by administering one or more short antisense compound. In a non-limiting example, the methods comprise the step of administering to an animal a therapeutically effective amount of a short antisense compound. Certain short antisense compounds inhibit the activity of GCCR and/ or inhibit expression of GCCR. In certain embodiments, the activity or expression of GCCR in a subject is inhibited by at least 10%, by at least 20%, by at least 25%, by at least 30%, by at least 40%, by at least 50%, by at least 60%, by at least 70%, by at least 75%, by at least 80%, by at least 85%, by at least 90%, by at least 95%, by at least 98%, by at least 99%, or by 100%. In certain embodiments, the activity or expression of GCCR in a subject is inhibited by at least 30%. In certain embodiments, the activity or expression of GCCR in a subject is inhibited by at least 50% or more.

The reduction of the expression of GCCR may be measured, for example, in blood, plasma, serum, adipose tissue, liver or any other body fluid, tissue or organ of the animal. In certain embodiments, cells contained within such fluids, tissues or organs being analyzed comprise nucleic acids encoding GCCR and/or they contain the GCCR protein itself.

Certain pharmaceutical and other compositions comprising short antisense compounds are also provided. In certain embodiments, short antisense compounds are be utilized in pharmaceutical compositions by adding to them an effective amount of a compound to a suitable pharmaceutically

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-109acceptable diluent or carrier.

In certain embodiments, short antisense compounds targeting a GCCR nucleic acid have any one or more properties or characteristics of the short antisense compounds generally described herein. In certain embodiments, short antisense compounds targeting a GCCR nucleic acid have a motif (wing deoxy gap-wing) selected from 1-12-1,1-1-10-2,2-10-1-1,3-10-3,2-10-3,2-10-2,1-10-1,1-10-2, 3-83, 2-8-2,1-8-1, 3-6-3 or 1-6-1,. In certain embodiments, short antisense compounds targeting a GCCR nucleic acid have a motif (wing - deoxy gap -wing) selected from 1-10-1,2-10-2,3-10-3, and 1-9-2.. In certain embodiments, short antisense compounds targeting a GCCR nucleic acid have a motif (wing deoxy gap -wing) selected from 3-10-3, 2-10-3, 2-10-2, 1-10-1,1-10-2, 2-8-2, 1-8-1, 3-6-3 or 1-6-1, more preferably 2-10-2 and 2-8-2.

In certain embodiments, provided herein are methods of treating an individual by administering one or more short antisense compound targeted to a GCCR nucleic acid or a pharmaceutical composition comprising such compound. Further provided are methods of treating a subject having a disease or conditions associated with GCCR activity by administering a short antisense compound targeted to a GCCR nucleic acid. In addition to diabetes, particularly type 2 diabetes, diseases and conditions associated with GCCR include but are not limited to, obesity, Metabolic syndrome X, Cushing's Syndrome, Addison's disease, inflammatory diseases such as asthma, rhinitis and arthritis, allergy, autoimmune disease, immunodeficiency, anorexia, cachexia, bone loss or bone frailty, and wound healing. Metabolic syndrome, metabolic syndrome X or simply Syndrome X refers to a cluster of risk factors that include obesity, dyslipidemia, particularly high blood triglycerides, glucose intolerance, high blood sugar and high blood pressure. In certain embodiments, short antisense compounds targeted to GCCR are used for amelioration of hyperglycemia induced by systemic steroid therapy. Moreover, antisense technology provides a means of inhibiting the expression of the glucocorticoid receptor β isoform, demonstrated to be overexpressed in patients refractory to glucocorticoid treatment.

In certain embodiments, the invention provides short antisense compounds targeted to a nucleic acid encoding GCGR, and which modulate the expression of glucocorticoid receptor. Pharmaceutical and other compositions comprising the compounds of the invention are also provided. Further provided are methods of screening for modulators of glucocorticoid receptor and methods of modulating the expression of glucocorticoid receptor in cells, tissues or animals comprising contacting said cells, tissues or animals with one or more of the compounds or compositions of the invention. Methods of treating an animal, particularly a human, suspected of having or being prone to a disease or condition associated with expression of glucocorticoid receptor are also set forth herein. Such methods comprise administering a therapeutically or prophylactically effective amount of one or more of the compounds or compositions of the invention to the person in need of treatment

Certain Short Antisense Compounds Targeted to a GCCR nucleic acid

In certain embodiments, short antisense compounds are targeted to a GCCR nucleic acid having

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-110the sequence of nucleotides 1 to 106000 of GENBANK® Accession No. AC012634, incorporated herein as SEQ ID NO: 8. In certain such embodiments, a short antisense compound targeted to SEQ ID NO: 8 is at least 90% complementary to SEQ ID NO: 8. In certain such embodiments, a short antisense compound targeted to SEQ ID NO: 8 is at least 95% complementary to SEQ ID NO: 8. In certain such embodiments, a short antisense compound targeted to SEQ ID NO: 8 is 100% complementary to SEQ ID NO: 8. In certain embodiments, a short antisense compound targeted to SEQ ID NO: 8 includes a nucleotide sequence selected from the nucleotide sequences set forth in Tables 10 and 11.

The nucleotide sequence set forth in each SEQ ID NO in Tables 10 and 11 is independent of any modification to a sugar moiety, an intemucleoside linkage, or a nucleobase. As such, short antisense compounds defined by a SEQ ID NO may comprise, independently, one or more modifications to a sugar moiety, an intemucleoside linkage, or a nucleobase. Short antisense compounds described by Isis Number (Isis NO.) indicate a combination of nucleobase sequence and one or more modifications to a sugar moiety, an intemucleoside linkage, or a nucleobase.

In certain embodiments, short antisense compounds targeted to a GCCR nucleic acid comprise a gapmer motif. In certain embodiments, a short antisense compound targeted to a GCCR nucleic acid comprises a 2-10-2 gapmer motif.

Tables 10 and 11 illustrate examples of short antisense compounds targeted to SEQ ID NO: 8. Table 10 illustrates short antisense compounds that are 100% complementary to SEQ ID NO: 8. Table 11 illustrates short antisense compounds that have one or two mismatches with respect to SEQ ID NO: 8. The column labeled ‘gapmer motif indicates the wing-gap-wing motif of each short antisense compounds. The gap segment comprises 2‘-deoxynucleotides and each nucleotide of each wing segment comprises a 2’-modified sugar. The particular 2’-modified sugar is also indicated in the ‘gapmer motif column. For example, ‘2-10-2 MOE’ means a 2-10-2 gapmer motif, where a gap segment of ten 2’deoxynucleotides is flanked by wing segments of two nucleotides, where the nucleotides of the wing segments are 2’-MOE nucleotides. Intemucleoside linkages are phosphorothioate. The short antisense compounds comprise 5-methylcytidine in place of unmodified cytosine, unless “unmodified cytosine” is listed in the gapmer motif column, in which case the indicated cytosines are unmodified cytosines. For example, “5-mC in gap only” indicates that the gap segment has 5-methylcytosines, while the wing segments have unmodified cytosines.

Table 10: Short Antisense Compounds targeted to SEQ ID NO: 8

<td> ISIS NO.</td><td> 5’ Target Site</td><td> 3’ Target Site</td><td> Sequence (5’-3’)</td><td> Gapmer Motif</td><td> SEQ ID NO</td>

<td> 371644</td><td> 88142</td><td> 88155</td><td> TTTGGGAGGTGGTC</td><td> 2-10-2 MOE</td><td> 413</td>

<td> 371645</td><td> 88156</td><td> 88169</td><td> CACACCAGGCAGAG</td><td> 2-10-2 MOE</td><td> 414</td>

<td> 371649</td><td> 88212</td><td> 88225</td><td> CTTTACAGCTTCCA</td><td> 2-10-2 MOE</td><td> 415</td>

<td> 371651</td><td> 88242</td><td> 88255</td><td> CACTACCTTCCACT</td><td> 2-10-2 MOE</td><td> 416</td>

<td> 371652</td><td> 88248</td><td> 88261</td><td> AACACACACTACCT</td><td> 2-10-2 MOE</td><td> 417</td>

<td> 371653</td><td> 88256</td><td> 88269</td><td> CTCTTCAAAACACA</td><td> 2-10-2 MOE</td><td> 418</td>

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<td> 371665</td><td> 92037</td><td> 92050</td><td> GTAATTGTGCTGTC</td><td> 2-10-2 MOE</td><td> 419</td>

<td> 371669</td><td> 92086</td><td> 92099</td><td> TTTTTCTTCGAATT</td><td> 2-10-2 MOE</td><td> 420</td>

<td> 371671</td><td> 92114</td><td> 92127</td><td> CATTTTCGATAGCG</td><td> 2-10-2 MOE</td><td> 421</td>

<td> 371673</td><td> 92142</td><td> 92155</td><td> ACCTTCCAGGTTCA</td><td> 2-10-2 MOE</td><td> 422</td>

Table 11: Short antisense compounds targeted to SEQ ID NO: 8 and having 1 or 2 mismatches

<td> ISIS NO</td><td> 5’ Target Site</td><td> 3’ Target Site</td><td> Sequence (5’-3’)</td><td> Gapmer Motif</td><td> SEQ ID NO</td>

<td> 371638</td><td> 2039</td><td> 2052</td><td> ATAGGAAGCATAAA</td><td> 2-10-2 MOE</td><td> 423</td>

<td> 371650</td><td> 4949</td><td> 4962</td><td> TCTTTTAAAGAAGA</td><td> 2-10-2 MOE</td><td> 424</td>

<td> 371673</td><td> 10187</td><td> 10200</td><td> ACCTTCCAGGTTCA</td><td> 2-10-2 MOE</td><td> 422</td>

<td> 371660</td><td> 13465</td><td> 13478</td><td> AAGGATATTTTAAA</td><td> 2-10-2 MOE</td><td> 425</td>

<td> 371660</td><td> 14428</td><td> 14441</td><td> AAGGATATTTTAAA</td><td> 2-10-2 MOE</td><td> 425</td>

<td> 371654</td><td> 15486</td><td> 15499</td><td> GAACAAAAATTAAA</td><td> 2-10-2 MOE</td><td> 427</td>

<td> 371661</td><td> 16638</td><td> 16651</td><td> TTCCACAGATCTGT</td><td> 2-10-2 MOE</td><td> 428</td>

<td> 371653</td><td> 17892</td><td> 17905</td><td> CTCTTCAAAACACA</td><td> 2-10-2 MOE</td><td> 418</td>

<td> 371679</td><td> 18444</td><td> 18457</td><td> TTTATAAAGTAAAG</td><td> 2-10-2 MOE</td><td> 429</td>

<td> 371645</td><td> 19816</td><td> 19829</td><td> CACACCAGGCAGAG</td><td> 2-10-2 MOE</td><td> 414</td>

<td> 371638</td><td> 21555</td><td> 21568</td><td> ATAGGAAGCATAAA</td><td> 2-10-2 MOE</td><td> 423</td>

<td> 371650</td><td> 21775</td><td> 21788</td><td> TCTTTTAAAGAAGA</td><td> 2-10-2 MOE</td><td> 424</td>

<td> 371679</td><td> 21902</td><td> 21915</td><td> TTTATAAAGTAAAG</td><td> 2-10-2 MOE</td><td> 429</td>

<td> 371655</td><td> 22507</td><td> 22520</td><td> TACTGTGAGAAATA</td><td> 2-10-2 MOE</td><td> 433</td>

<td> 371655</td><td> 22722</td><td> 22735</td><td> TACTGTGAGAAATA</td><td> 2-10-2 MOE</td><td> 433</td>

<td> 371672</td><td> 25662</td><td> 25675</td><td> TTCCAGCTTGAAGA</td><td> 2-10-2 MOE</td><td> 435</td>

<td> 371678</td><td> 25926</td><td> 25939</td><td> GATCAGTTCTCATG</td><td> 2-10-2 MOE</td><td> 436</td>

<td> 371655</td><td> 26041</td><td> 26054</td><td> TACTGTGAGAAATA</td><td> 2-10-2 MOE</td><td> 433</td>

<td> 371638</td><td> 29770</td><td> 29783</td><td> ATAGGAAGCATAAA</td><td> 2-10-2 MOE</td><td> 423</td>

<td> 371668</td><td> 30551</td><td> 30564</td><td> TTATCAATGATGCA</td><td> 2-10-2 MOE</td><td> 439</td>

<td> 371670</td><td> 40584</td><td> 40597</td><td> GCATGCTGGACAGT</td><td> 2-10-2 MOE</td><td> 440</td>

<td> 371654</td><td> 43331</td><td> 43344</td><td> GAACAAAAATTAAA</td><td> 2-10-2 MOE</td><td> 427</td>

<td> 371650</td><td> 46024</td><td> 46037</td><td> TCTTTTAAAGAAGA</td><td> 2-10-2 MOE</td><td> 424</td>

<td> 371659</td><td> 50372</td><td> 50385</td><td> TTGCACCTGAACTA</td><td> 2-10-2 MOE</td><td> 443</td>

<td> 371634</td><td> 50565</td><td> 50578</td><td> CAGAATATATTTCT</td><td> 2-10-2 MOE</td><td> 444</td>

<td> 371673</td><td> 56942</td><td> 56955</td><td> ACCTTCCAGGTTCA</td><td> 2-10-2 MOE</td><td> 422</td>

<td> 371654</td><td> 62372</td><td> 62385</td><td> GAACAAAAATTAAA</td><td> 2-10-2 MOE</td><td> 427</td>

<td> 371679</td><td> 63537</td><td> 63550</td><td> TTTATAAAGTAAAG</td><td> 2-10-2 MOE</td><td> 429</td>

<td> 371654</td><td> 64908</td><td> 64921</td><td> GAACAAAAATTAAA</td><td> 2-10-2 MOE</td><td> 427</td>

<td> 371661</td><td> 65795</td><td> 65808</td><td> TTCCACAGATCTGT</td><td> 2-10-2 MOE</td><td> 428</td>

<td> 371645</td><td> 70997</td><td> 71010</td><td> CACACCAGGCAGAG</td><td> 2-10-2 MOE</td><td> 414</td>

<td> 371661</td><td> 77400</td><td> 77413</td><td> TTCCACAGATCTGT</td><td> 2-10-2 MOE</td><td> 428</td>

<td> 371663</td><td> 82329</td><td> 82342</td><td> ATAAGAGATTAAAA</td><td> 2-10-2 MOE</td><td> 450</td>

<td> 371633</td><td> 83426</td><td> 83439</td><td> TCCCCCTTCTCATT</td><td> 2-10-2 MOE</td><td> 451</td>

<td> 371662</td><td> 85873</td><td> 85886</td><td> GGGCATTGTTAAAA</td><td> 2-10-2 MOE</td><td> 452</td>

<td> 371654</td><td> 86476</td><td> 86489</td><td> GAACAAAAATTAAA</td><td> 2-10-2 MOE</td><td> 427</td>

<td> 371679</td><td> 86516</td><td> 86529</td><td> TTTATAAAGTAAAG</td><td> 2-10-2 MOE</td><td> 429</td>

<td> 371641</td><td> 88097</td><td> 88110</td><td> AGAACTCACATCTG</td><td> 2-10-2 MOE</td><td> 455</td>

<td> 371642</td><td> 88111</td><td> 88124</td><td> GAGCTGGACGGAGG</td><td> 2-10-2 MOE</td><td> 456</td>

<td> 371646</td><td> 88170</td><td> 88183</td><td> AAGCTTCATCGGAG</td><td> 2-10-2 MOE</td><td> 457</td>

<td> 371647</td><td> 88184</td><td> 88197</td><td> ATAATGGCATCCCG</td><td> 2-10-2 MOE</td><td> 458</td>

<td> 371650</td><td> 88226</td><td> 88239</td><td> TCTTTTAAAGAAGA</td><td> 2-10-2 MOE</td><td> 424</td>

<td> 371673</td><td> 91493</td><td> 91506</td><td> ACCTTCCAGGTTCA</td><td> 2-10-2 MOE</td><td> 422</td>

<td> 371664</td><td> 92030</td><td> 92043</td><td> TGCTGTCCTATAAG</td><td> 2-10-2 MOE</td><td> 460</td>

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<td> 371666</td><td> 92044</td><td> 92057</td><td> CACAAAGGTAATTG</td><td> 2-10-2 MOE</td><td> 461</td>

<td> 371667</td><td> 92058</td><td> 92071</td><td> ATCATTTCTTCCAG</td><td> 2-10-2 MOE</td><td> 462</td>

<td> 371668</td><td> 92072</td><td> 92085</td><td> TTATCAATGATGCA</td><td> 2-10-2 MOE</td><td> 463</td>

<td> 371670</td><td> 92100</td><td> 92113</td><td> GCATGCTGGACAGT</td><td> 2-10-2 MOE</td><td> 440</td>

<td> 371672</td><td> 92128</td><td> 92141</td><td> TTCCAGCTTGAAGA</td><td> 2-10-2 MOE</td><td> 435</td>

<td> 371674</td><td> 92147</td><td> 92160</td><td> CCATTACCTTCCAG</td><td> 2-10-2 MOE</td><td> 466</td>

<td> 371637</td><td> 92983</td><td> 92996</td><td> GCATAAACAGGGTT</td><td> 2-10-2 MOE</td><td> 467</td>

<td> 371654</td><td> 93928</td><td> 93941</td><td> GAACAAAAATTAAA</td><td> 2-10-2 MOE</td><td> 427</td>

<td> 371641</td><td> 99772</td><td> 99785</td><td> AGAACTCACATCTG</td><td> 2-10-2 MOE</td><td> 455</td>

<td> 371679</td><td> 99883</td><td> 99896</td><td> TTTATAAAGTAAAG</td><td> 2-10-2 MOE</td><td> 429</td>

<td> 371660</td><td> 99933</td><td> 99946</td><td> AAGGATATTTTAAA</td><td> 2-10-2 MOE</td><td> 425</td>

<td> 371635</td><td> 105004</td><td> 105017</td><td> TATGAAAGGAATGT</td><td> 2-10-2 MOE</td><td> 472</td>

<td> 371654</td><td> 105028</td><td> 105041</td><td> GAACAAAAATTAAA</td><td> 2-10-2 MOE</td><td> 427</td>

<td> 371676</td><td> 106482</td><td> 106495</td><td> TTCCTTAAGCTTCC</td><td> 2-10-2 MOE</td><td> 474</td>

<td> 371650</td><td> 107838</td><td> 107851</td><td> TCTTTTAAAGAAGA</td><td> 2-10-2 MOE</td><td> 424</td>

<td> 371673</td><td> 110922</td><td> 110935</td><td> ACCTTCCAGGTTCA</td><td> 2-10-2 MOE</td><td> 422</td>

<td> 371673</td><td> 111580</td><td> 111593</td><td> ACCTTCCAGGTTCA</td><td> 2-10-2 MOE</td><td> 422</td>

<td> 371634</td><td> 114608</td><td> 114621</td><td> CAGAATATATTTCT</td><td> 2-10-2 MOE</td><td> 444</td>

<td> 371638</td><td> 115040</td><td> 115053</td><td> ATAGGAAGCATAAA</td><td> 2-10-2 MOE</td><td> 423</td>

<td> 371660</td><td> 116244</td><td> 116257</td><td> AAGGATATTTTAAA</td><td> 2-10-2 MOE</td><td> 425</td>

<td> 371663</td><td> 116657</td><td> 116670</td><td> ATAAGAGATTAAAA</td><td> 2-10-2 MOE</td><td> 450</td>

<td> 371673</td><td> 118068</td><td> 118081</td><td> ACCTTCCAGGTTCA</td><td> 2-10-2 MOE</td><td> 422</td>

<td> 371666</td><td> 118834</td><td> 118847</td><td> CACAAAGGTAATTG</td><td> 2-10-2 MOE</td><td> 461</td>

<td> 371660</td><td> 119858</td><td> 119871</td><td> AAGGATATTTTAAA</td><td> 2-10-2 MOE</td><td> 425</td>

<td> 371660</td><td> 120210</td><td> 120223</td><td> AAGGATATTTTAAA</td><td> 2-10-2 MOE</td><td> 425</td>

<td> 371662</td><td> 120876</td><td> 120889</td><td> GGGCATTGTTAAAA</td><td> 2-10-2 MOE</td><td> 452</td>

<td> 371655</td><td> 124004</td><td> 124017</td><td> TACTGTGAGAAATA</td><td> 2-10-2 MOE</td><td> 433</td>

<td> 371656</td><td> 124170</td><td> 124183</td><td> GAACAGTTAAACAT</td><td> 2-10-2 MOE</td><td> 485</td>

In certain embodiments, a target region is nucleotides 88142-88269 of SEQ ID NO: 8. In certain embodiments, a short antisense compound is targeted to nucleotides 88142-88269 of SEQ ID NO: 8. In certain such embodiments, a short antisense compound targeted to nucleotides 88142-88269 comprises a 5 nucleotide sequence selected from SEQ ID NO 413, 414, 415, 416, 417, or 418. In certain such embodiments, an antisense compound targeted to nucleotides 88142-88269 of SEQ ID NO: 8 is selected from Isis NO. 371644,371645,371649,371651,371652, or 371653.

In certain embodiments, a target region is nucleotides 88142-88169 of SEQ ID NO: 8. In certain embodiments, a short antisense compound is targeted to nucleotides 88142-88169 of SEQ ID NO: 8. In 10 certain such embodiments, a short antisense compound targeted to nucleotides 88142-88169 comprises a nucleotide sequence selected from SEQ ID NO 413 or 414. In certain such embodiments, an antisense compound targeted to nucleotides 88142-88169 of SEQ ID NO: 8 is selected from Isis NO. 371644 or 371645.

In certain embodiments, a target region is nucleotides 88242-88269 of SEQ ID NO: 8. In certain 15 embodiments, a short antisense compound is targeted to nucleotides 88242-88269 of SEQ ID NO: 8. In certain such embodiments, a short antisense compound targeted to nucleotides 88242-88269 comprises a nucleotide sequence selected from SEQ ID NO 416, 417, or 418. In certain such embodiments, an antisense compound targeted to nucleotides 88242-88269 of SEQ ID NO: 8 is selected from Isis NO.

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-113371651,371652, or 371653.

In certain embodiments, a target region is nucleotides 92037-92155 of SEQ ID NO; 8. In certain embodiments, a short antisense compound is targeted to nucleotides 92037-92155 of SEQ ID NO: 8. In certain such embodiments, a short antisense compound targeted to nucleotides 92037-92155 comprises a nucleotide sequence selected from SEQ ID NO 419, 420,421, or 422. In certain such embodiments, an antisense compound targeted to nucleotides 92037-92155 of SEQ ID NO: 8 is selected from Isis NO. 371665,371669,371671, or 171673.

In certain embodiments, a target region is nucleotides 92114-92155 of SEQ ID NO: 8. In certain embodiments, a short antisense compound is targeted to nucleotides 92114-92155 of SEQ ID NO: 8. In certain such embodiments, a short antisense compound targeted to nucleotides 92114-92155 comprises a nucleotide sequence selected from SEQ ID NO 421 or 422. In certain such embodiments, an antisense compound targeted to nucleotides 92114-92155 of SEQ ID NO: 8 is selected from Isis NO. 371671 or 171673.

In certain embodiments, short antisense compounds targeted to a GCCR. nucleic acid are 8 to 16, preferably 9 to 15, more preferably 9 to 14, more preferably 10 to 14 nucleotides in length. In certain embodiments, short antisense compounds targeted to a GCCR nucleic acid are 9 to 14 nucleotides in length. In certain embodiments, short antisense compounds targeted to a GCCR nucleic acid are 10 to 14 nucleotides in length. In certain embodiments, such short antisense compounds are short antisense oligonucleotides.

In certain embodiments, short antisense compounds targeted to a GCCR nucleic acid are short gapmers. In certain such embodiments, short gapmers targeted to a GCCR nucleic acid comprise at least one high affinity modification in one or more wings of the compound. In certain embodiments, short antisense compounds targeted to a GCCR nucleic acid comprise 1 to 3 high-affinity modifications in each wing. In certain such embodiments, the nucleosides or nucleotides of the wing comprise a 2’ modification. In certain such embodiments, the monomers of the wing are BNA’s. In certain such embodiments, the monomers of the wing are selected from α-L-Mcthyleneoxy (4’-CH₂-O-2’) BNA, β-DMethyleneoxy (4’-CH₂-O-2’) BNA , Ethyleneoxy (4’&lt;CH₂)₂O-2’) BNA , Aminooxy (4’-CH₂-O-N(R)2’) BNA and Oxyamino (4’-CH₂-N(R)-O-2’) BNA. In certain embodiments, the monomers of a wing comprise a substituent at the 2’ position selected from allyl, amino, azido, thio, O-allyl, O-C|-C|₀ alkyl, OCFj, O-(CH₂)₂-O-CH₃, 2'-O(CH₂)₂SCH₃,O-(ClI₂)₂-O-N(R_m)(R_u), and O-CH₂-C(=O)-N(R_m)(R^), where each Rn, and Rfl is, independently, H or substituted or unsubstituted C_rCi₀ alkyl. In certain embodiments, the monomers of a wing are 2’MOE nucleotides.

In certain embodiments, short antisense compounds targeted to a GCCR nucleic acid comprise a gap between the 5’ wing and the 3’ wing. In certain embodiments the gap comprises five, six, seven, eight, nine, ten, eleven, twelve, thirteen, or fourteen monomers. In certain embodiments, the monomers of the gap are unmodified deoxyribonucleotides. In certain embodiments, the monomers of the gap are unmodified ribonucleotides. In certain embodiments, gap modifications (if any) gap result in an

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-114antisense compound that, when bound to its target nucleic acid, supports cleavage by an RNase, including, but not limited to, RNase H.

In certain embodiments, short antisense compounds targeted to a GCCR nucleic acid have unifonn monomeric linkages. In certain such embodiments, those linkages are all phosphorothioate linkages. In certain embodiments, the linkages are all phosphodiester linkages. In certain embodiments, short antisense compounds targeted to a GCCR nucleic acid have mixed backbones.

In certain embodiments, short antisense compounds targeted to a GCCR nucleic acid are 8 monomers in length. In certain embodiments, short antisense compounds targeted to a GCCR nucleic acid are 9 monomers in length. In certain embodiments, short antisense compounds targeted to a GCCR nucleic acid are 10 monomers in length. In certain embodiments, short antisense compounds targeted to a GCCR nucleic acid are 11 monomers in length. In certain embodiments, short antisense compounds targeted to a GCCR nucleic acid are monomers in length. In certain embodiments, short antisense compounds targeted to a GCCR nucleic acid are 13 monomers in length. In certain embodiments, short antisense compounds targeted to a GCCR nucleic acid are 14 monomers in length. In certain embodiments, short antisense compounds targeted to a GCCR nucleic acid are 15 monomers in length. In certain embodiments, short antisense compounds targeted to a GCCR nucleic acid are 16 monomers in length. In certain embodiments, short antisense compounds targeted to a GCCR nucleic acid comprise 9 to 15 monomers. In certain embodiments, short antisense compounds targeted to a GCCR nucleic acid comprise 10 to 15 monomers. In certain embodiments, short antisense compounds targeted to a GCCR nucleic acid comprise 12 to 14 monomers. In certain embodiments, short antisense compounds targeted to a GCCR nucleic acid comprise 12 to 14 nucleotides or nucleosides.

In certain embodiments, the invention provides methods of modulating expression of GCCR. In certain embodiments, such methods comprise use of one or more short antisense compound targeted to a GCCR nucleic acid, wherein the short antisense compound targeted to a GCCR nucleic acid is from about 8 to about 16, preferably 9 to 15, more preferably 9 to 14, more preferably 10 to 14 monomers (i.e. from about 8 to about 16 linked monomers). One of ordinary skill in the art will appreciate that this comprehends methods of modulating expression of GCCR using one or more short antisense compounds targeted to a GCCR nucleic acid of 8,9,10,11,12,13,14,15 or 16 monomers.

In certain embodiments, methods of modulating GCCR comprise use of a short antisense compound targeted to a GCCR nucleic acid that is 8 monomers in length. In certain embodiments, methods of modulating GCCR comprise use of a short antisense compound targeted to a GCCR nucleic acid that is 9 monomers in length. In certain embodiments, methods of modulating GCCR comprise use of a short antisense compound targeted to a GCCR nucleic acid that is 10 monomers in length. In certain embodiments, methods of modulating GCCR comprise use of a short antisense compound targeted to a GCCR nucleic acid that is 11 monomers in length. In certain embodiments, methods of modulating GCCR comprise use of a short antisense compound targeted to a GCCR nucleic acid that is 12 monomers in length. In certain embodiments, methods of modulating GCCR comprise use of a short antisense

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-115compound targeted to a GCCR nucleic acid that is 13 monomers in length. In certain embodiments, methods of modulating GCCR comprise use of a short antisense compound targeted to a GCCR nucleic acid that is 14 monomers in length. In certain embodiments, methods of modulating GCCR comprise use of a short antisense compound targeted to a GCCR nucleic acid that is 15 monomers in length. In certain embodiments, methods of modulating GCCR comprise use of a short antisense compound targeted to a GCCR nucleic acid that is 16 monomers in length.

In certain embodiments, methods of modulating expression of GCCR comprise use of a short antisense compound targeted to a GCCR nucleic acid comprising 9 to 15 monomers. In certain embodiments, methods of modulating expression of GCCR comprise use of a short antisense compound targeted to a GCCR nucleic acid comprising 10 to 15 monomers. In certain embodiments, methods of modulating expression of GCCR comprise use of a short antisense compound targeted to a GCCR nucleic acid comprising 12 to 14 monomers. In certain embodiments, methods of modulating expression of GCCR comprise use of a short antisense compound targeted to a GCCR nucleic acid comprising 12 or 14 nucleotides or nucleosides.

7. Glucagon Receptor (GCGR)

The maintenance of normal glycemia is a carefully regulated metabolic event. Glucagon, the 29amino acid peptide responsible for maintaining blood glucose levels in the postabsorbative state, increases glucose release from the liver by activating hepatic glycogenolysis, gluconeogenesis, stimulating lipolysis in adipose tissue, and stimulating insulin secretion. During high blood glucose levels, insulin reverses the glucagon-mediated enhancement of glycogenolysis and gluconeogenesis. In patients with diabetes, insulin is either not available or not fully effective. While treatment for diabetes has traditionally focused on increasing insulin levels, antagonism of glucagon function has been considered as an alternative therapy. As glucagon exerts its physiological effects by signaling through the glucagon receptor, the glucagon receptor has been proposed as a potential therapeutic target for diabetes (Madsen et al., Curr. Pharm. Des., 1999,5,683-691).

Glucagon receptor is belongs to the superfamily of G-protein-coupled receptors having seven transmembrane domains. It is also a member of the smaller sub-family of homologous receptors which bind peptides that are structurally similar to glucagon. The gene encoding human glucagon receptor was cloned in 1994 and analysis of the genomic sequence revealed multiple introns and an 82% identity to the rat glucagon receptor gene (Lok et al., Gene, 1994, 140, 203-209.; MacNeil et al., Biochem. Biophys. Res. Commun., 1994, 198, 328-334). Cloning of the rat glucagon receptor gene also led to the description of multiple alternative splice variants (Maget et al., FEBS Lett., 1994, 351, 271-275). The human glucagon receptor gene is localized to chromosome 17q25 (Menzel et al., Genomics, 1994, 20, 327-328). A missense mutation of Gly to Ser at codon 40 in the glucagon receptor gene leads to a 3-fold lower affinity for glucagon (Fujisawa et al., Diabetologia, 1995, 38,983-985) and this mutation has been linked to several disease states, including non-insulin-dependent diabetes mellitus (Fujisawa et al.,

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-116Diabetologia, 1995, 38, 983-985), hypertension (Chambers and Morris, Nat. Genet., 1996, 12, 122), and central adiposity (Siani et al., Obes. Res., 2001,9,722-726).

Definitions “Glucagon receptor” is the gene product or protein of which expression is to be modulated by administration of a short antisense compound. Glucagon receptor is generally referred to as GCGR but may also be referred to as GR, GGR, MGC138246, MGC93090.

“GCGR nucleic acid means any nucleic acid encoding GCGR. For example, in certain embodiments, a GCGR nucleic acid includes, without limitation, a GCGR sequence encoding GCGR, an RNA sequence transcribed from DNA encoding GCGR, and an mRNA sequence encoding GCGR. “GCGR mRNA” means an mRNA encoding a GCGR protein.

Therapeutic Indications

Antisense technology is an effective means for reducing glucagon receptor (GCGR) expression and has proven to be uniquely useful in a number of therapeutic, diagnostic, and research applications. As such, in certain embodiments, the present invention provides short antisense compounds targeted to a nucleic acid encoding glucagon receptor, and which modulate the expression of glucagon receptor. Further provided herein are short antisense compounds capable of inhibiting GCGR expression. Also provided herein are methods of treating an individual comprising administering one or more pharmaceutical compositions comprising a short antisense compound targeted to a GCGR nucleic acid. In certain embodiments, because short antisense compounds targeted to a GCGR nucleic acid inhibit GCGR expression, provided herein are methods of treating a subject having a disease or condition associated with GCGR activity by administering one or more pharmaceutical compositions comprising a short antisense compound targeted to a GCGR nucleic acid. For example, provided herein are methods of treating a subject having high blood glucose, hyperglycemia, prediabetes, diabetes, Type 2 diabetes, metabolic syndrome, obesity and/or insulin resistance.

Also contemplated herein are pharmaceutical composition comprising one or more short antisense compounds targeted to GCGR and optionally a pharmaceutically acceptable carrier, diluent, enhancer or excipient. Certain compounds of the invention can also be used in the manufacture of a medicament for the treatment of diseases and disorders related to glucagon effects mediated by GCGR.

Certain embodiments of the present invention include methods of reducing the expression of GCGR in tissues or cells comprising contacting said cells or tissues with a short antisense compound targeted to a nucleic acid encoding GCGR or pharmaceutical composition comprising such a short antisense compound. In certain such embodiments, the invention provides methods of decreasing blood glucose levels, blood triglyceride levels, or blood cholesterol levels in a subject comprising administering to the subject a short antisense compound or a pharmaceutical composition. Blood levels may be plasma levels or serum levels. Also contemplated are methods of improving insulin sensitivity, methods of

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-117increasing GLP-1 levels and methods of inhibiting hepatic glucose output in an animal comprising administering to said animal an antisense oligonucleotide or a pharmaceutical composition of the invention. An improvement in insulin sensitivity may be indicated by a reduction in circulating insulin levels.

In certain embodiments, the invention provides methods of treating a subject having a disease or condition associated with glucagon activity via GCGR comprising administering to the subject a therapeutically or prophylactically effective amount of a short antisense compound or a pharmaceutical composition. In certain embodiments, such disease or condition may be a metabolic disease or condition. In certain embodiments, the metabolic disease or condition is diabetes, hyperglycemia, hyperlipidemia, metabolic syndrome X, obesity, primary hyperglucagonemia, insulin deficiency, or insulin resistance. In some embodiments, the diabetes is Type 2 diabetes. In some embodiments the obesity is diet-induced. In some embodiments, hyperlipidemia is associated with elevated blood lipid levels. Lipids include cholesterol and triglycerides. In one embodiment, the condition is liver steatosis. In some embodiments, the steatosis is stcatohepatitis or non-alcoholic steatohepatitis.

In certain embodiments, the invention provides methods of preventing or delaying the onset of elevated blood glucose levels in an animal as well as methods of preserving beta-cell function in an animal using the oligomeric compounds delineated herein.

Certain short antisense compounds targeted to GCGR can be used to modulate the expression of GCGR in a subject in need thereof, such as an animal, including, but not limited to, a human In certain embodiments, such methods comprise the step of administering to said animal an effective amount of a short antisense compound that reduces expression of GCGR RNA. In certain embodiments, short antisense compounds effectively reduce the levels or function of GCGR RNA. Because reduction in GCGR mRNA levels can lead to alteration in GCGR protein products of expression as well, such resultant alterations can also be measured. Certain antisense compounds that effectively reduce the levels or function of GCGR RNA or protein products of expression is considered an active antisense compound. In certain embodiments, short antisense compounds reduce die expression of GCGR causing a reduction of RNA by at least 10%, by at least 20%, by at least 25%, by at least 30%, by at least 40%, by at least 50%, by at least 60%, by at least 70%, by at least 75%, by at least 80%, by at least 85%, by at least 90%, by at least 95%, by at least 98%, by at least 99%, or by 100%.

Further provided are methods of screening for modulators of glucagon receptor and methods of modulating the expression of glucagon receptor in cells, tissues or animals comprising contacting said cells, tissues or animals with one or more short antisense compounds targeted to GCGR or with compositions comprising such compounds. Methods of treating an animal, particularly a human, suspected of having or being prone to a disease or condition associated with expression of glucagon receptor are also set forth herein. Certain such methods comprise administering a therapeutically or prophylactically effective amount of one or more of the compounds or compositions of the invention to the person in need of treatment.

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-118The reduction of the expression of glucagon receptor may be measured, for example, in blood, plasma, serum, adipose tissue, liver or any other body fluid, tissue or organ of the animal. Preferably, the cells contained within said fluids, tissues or organs being analyzed contain a nucleic acid molecule encoding glucagon receptor protein and/or the glucagon receptor protein itself.

Pharmaceutical and other compositions comprising short antisense compounds are also provided. In certain embodiments short antisense compounds targeted to a nucleic acid encoding GCGR are utilized in pharmaceutical compositions by adding an effective amount of a compound to a suitable pharmaceutically acceptable diluent or carrier.

The short antisense compounds targeting a GCGR nucleic acid may have any one or more properties or characteristics of the short antisense compounds generally described herein. In certain embodiments, short antisense compounds targeting a GCGR nucleic acid have a motif (wing - deoxy gap -wing) selected from 1-12-1,1-1-10-2,2-10-1-1,3-10-3,2-10-3,2-10-2,1-10-1,1-10-2, 3-8-3,2-8-2,18-1, 3-6-3 or 1-6-1. In certain embodiments, short antisense compounds targeting a GCGR nucleic acid have a motif (wing- deoxy gap -wing) selected from 1-12-1,2-10-2,3-10-3,3-8-3,1-1-10-2.

Certain Short Antisense Compounds Targeted to a GCGR nucleic acid

In certain embodiments, short antisense compounds are targeted to a GCGR nucleic acid having the sequence GENBANK® Accession No. NM 000160.1, incorporated herein as SEQ ID NO: 9. In certain such embodiments, a short antisense compound targeted to SEQ ID NO: 9 is at least 90% complementary to SEQ ID NO: 9. In certain such embodiments, a short antisense compound targeted to SEQ ID NO: 9 is at least 95% complementary to SEQ ID NO: 9. In certain such embodiments, a short antisense compound^targeted to SEQ ID NO: 9 is 100% complementary to SEQ ID NO: 9. In certain embodiments, a short antisense compound targeted to SEQ ID NO: 9 includes a nucleotide sequence selected from the nucleotide sequences set forth in Tables 12 and 13.

The nucleotide sequences set forth in each SEQ ID NO in Tables 12 and 13 are independent of any modification to a sugar moiety, an intemucleoside linkage, or a nucleobase. As such, short antisense compounds defined by a SEQ ID NO may comprise, independently, one or more modifications to a sugar moiety, an intemucleoside linkage, or a nucleobase. Short antisense compounds described by Isis Number (Isis NO.) indicate a combination of nucleobase sequence and one or more modifications to a sugar moiety, an intemucleoside linkage, or a nucleobase.

In certain embodiments, short antisense compounds targeted to a GCCR nucleic acid comprise a gapmer motif. In certain embodiments, a short antisense compound targeted to a GCCR nucleic acid comprises a 3-10-3 gapmer motif. In certain embodiments, short antisense compounds targeted to a GCCR nucleic acid comprise a gapmer motif. In certain embodiments, a short antisense compound targeted to a GCCR nucleic acid comprises a 3-8-3 gapmer motif. In certain embodiments, short antisense compounds targeted to a GCCR nucleic acid comprise a gapmer motif. In certain embodiments, a short antisense compound targeted to a GCCR nucleic acid comprises a 2-10-2 gapmer motif.

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-119Tables 12 and 13 illustrate examples of short antisense compounds targeted to SEQ ID NO: 9. Table 12 illustrates short antisense compounds that are 100% complementary to SEQ ID NO: 9. Table 13 illustrates short antisense compounds that have one or two mismatches with respect to SEQ ID NO: 9. The column labeled 'gapmer motif indicates the wing-gap-wing motif of each short antisense compounds. The gap segment comprises 2’-deoxynucleotides and each nucleotide of each wing segment comprises a 2’-modified sugar. The particular 2’-modified sugar is also indicated in the ‘gapmer motif column. For example, ‘2-10-2 MOE’ means a 2-10-2 gapmer motif, where a gap segment of ten 2’deoxynucleotides is flanked by wing segments of two nucleotides, where the nucleotides of the wing segments are 2’-MOE nucleotides. Intemucleoside linkages are phosphorothioate. The short antisense compounds comprise 5-methylcytidine in place of unmodified cytosine, unless “unmodified cytosine” is listed in the gapmer motif column, in which case the indicated cytosines are unmodified cytosines. For example, “5-mC in gap only” indicates that the gap segment has 5-methylcytosines, while the wing segments have unmodified cytosines.

Table 12: Short Antisense Componnds targeted to SEQ ID NO: 9

<td> ISIS NO.</td><td> 5’ Target Site</td><td> 3’ Target Site</td><td> Sequence (5’-3’)</td><td> Gapmer Motif</td><td> SEQ ID NO</td>

<td> 338463</td><td> 378</td><td> 393</td><td> TAGAGCTTCCACTTCT</td><td> 3-10-3 MOE</td><td> 486</td>

<td> 338534</td><td> 378</td><td> 391</td><td> GAGCTTCCACTTCT</td><td> 3-8-3 MOE</td><td> 487</td>

<td> 327130</td><td> 499</td><td> 512</td><td> TGTTGGCCGTGGTA</td><td> 3-8-3 MOE</td><td> 488</td>

<td> 327131</td><td> 500</td><td> 513</td><td> ATGTTGGCCGTGGT</td><td> 3-8-3 MOE</td><td> 489</td>

<td> 327132</td><td> 501</td><td> 514</td><td> GATGTTGGCCGTGG</td><td> 3-8-3 MOE</td><td> 490</td>

<td> 327133</td><td> 502</td><td> 515</td><td> AGATGTTGGCCGTG</td><td> 3-8-3 MOE</td><td> 491</td>

<td> 327134</td><td> 503</td><td> 516</td><td> GAGATGTTGGCCGT</td><td> 3-8-3 MOE</td><td> 492</td>

<td> 327135</td><td> 504</td><td> 517</td><td> GGAGATGTTGGCCG</td><td> 3-8-3 MOE</td><td> 493</td>

<td> 327136</td><td> 505</td><td> 518</td><td> AGGAGATGTTGGCC</td><td> 3-8-3 MOE</td><td> 494</td>

<td> 327137</td><td> 506</td><td> 519</td><td> CAGGAGATGTTGGC</td><td> 3-8-3 MOE</td><td> 495</td>

<td> 327138</td><td> 507</td><td> 520</td><td> GCAGGAGATGTTGG</td><td> 3-8-3 MOE</td><td> 496</td>

<td> 327139</td><td> 508</td><td> 521</td><td> GGCAGGAGATGTTG</td><td> 3-8-3 MOE</td><td> 497</td>

<td> 327140</td><td> 531</td><td> 544</td><td> GTGGTGCCAAGGCA</td><td> 3-8-3 MOE</td><td> 498</td>

<td> 327141</td><td> 532</td><td> 545</td><td> TGTGGTGCCAAGGC</td><td> 3-8-3 MOE</td><td> 499</td>

<td> 327142</td><td> 533</td><td> 546</td><td> TTGTGGTGCCAAGG</td><td> 3-8-3 MOE</td><td> 500</td>

<td> 327143</td><td> 534</td><td> 547</td><td> TTTGTGGTGCCAAG</td><td> 3-8-3 MOE</td><td> 501</td>

<td> 327144</td><td> 535</td><td> 548</td><td> CTTTGTGGTGCCAA</td><td> 3-8-3 MOE</td><td> 502</td>

<td> 327145</td><td> 536</td><td> 549</td><td> ACTTTGTGGTGCCA</td><td> 3-8-3 MOE</td><td> 503</td>

<td> 327146</td><td> 537</td><td> 550</td><td> CACTTTGTGGTGCC</td><td> 3-8-3 MOE</td><td> 504</td>

<td> 327147</td><td> 538</td><td> 551</td><td> GCACTTTGTGGTGC</td><td> 3-8-3 MOE</td><td> 505</td>

<td> 327148</td><td> 539</td><td> 552</td><td> TGCACTTTGTGGTG</td><td> 3-8-3 MOE</td><td> 506</td>

<td> 327149</td><td> 540</td><td> 553</td><td> TTGCACTTTGTGGT</td><td> 3-8-3 MOE</td><td> 507</td>

<td> 327150</td><td> 545</td><td> 558</td><td> CGGTGTTGCACTTT</td><td> 3-8-3 MOE</td><td> 508</td>

<td> 327151</td><td> 546</td><td> 559</td><td> GCGGTGTTGCACTT</td><td> 3-8-3 MOE</td><td> 509</td>

<td> 327152</td><td> 547</td><td> 560</td><td> AGCGGTGTTGCACT</td><td> 3-8-3 MOE</td><td> 510</td>

<td> 327153</td><td> 548</td><td> 561</td><td> AAGCGGTGTTGCAC</td><td> 3-8-3 MOE</td><td> 511</td>

<td> 327154</td><td> 549</td><td> 562</td><td> GAAGCGGTGTTGCA</td><td> 3-8-3 MOE</td><td> 512</td>

<td> 327155</td><td> 550</td><td> 563</td><td> CGAAGCGGTGTTGC</td><td> 3-8-3 MOE</td><td> 513</td>

<td> 327156</td><td> 551</td><td> 564</td><td> ACGAAGCGGTGTTG</td><td> 3-8-3 MOE</td><td> 514</td>

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<td> 327157</td><td> 552</td><td> 565</td><td> CACGAAGCGGTGTT</td><td> 3-8-3 MOE</td><td> 515</td>

<td> 327158</td><td> 553</td><td> 566</td><td> ACACGAAGCGGTGT</td><td> 3-8-3 MOE</td><td> 516</td>

<td> 327159</td><td> 554</td><td> 567</td><td> AACACGAAGCGGTG</td><td> 3-8-3 MOE</td><td> 517</td>

<td> 345897</td><td> 684</td><td> 697</td><td> GCTGCTGTACATCT</td><td> 2-10-2 MOE</td><td> 518</td>

<td> 327160</td><td> 684</td><td> 697</td><td> GCTGCTGTACATCT</td><td> 3-8-3 MOE</td><td> 518</td>

<td> 327161</td><td> 685</td><td> 698</td><td> AGCTGCTGTACATC</td><td> 3-8-3 MOE</td><td> 520</td>

<td> 327162</td><td> 686</td><td> 699</td><td> AAGCTGCTGTACAT</td><td> 3-8-3 MOE</td><td> 521</td>

<td> 327163</td><td> 687</td><td> 700</td><td> GAAGCTGCTGTACA</td><td> 3-8-3 MOE</td><td> 522</td>

<td> 327164</td><td> 688</td><td> 701</td><td> GGAAGCTGCTGTAC</td><td> 3-8-3 MOE</td><td> 523</td>

<td> 327165</td><td> 689</td><td> 702</td><td> TGGAAGCTGCTGTA</td><td> 3-8-3 MOE</td><td> 524</td>

<td> 327166</td><td> 690</td><td> 703</td><td> CTGGAAGCTGCTGT</td><td> 3-8-3 MOE</td><td> 525</td>

<td> 327167</td><td> 691</td><td> 704</td><td> CCTGGAAGCTGCTG</td><td> 3-8-3 MOE</td><td> 526</td>

<td> 327168</td><td> 692</td><td> 705</td><td> ACCTGGAAGCTGCT</td><td> 3-8-3 MOE</td><td> 527</td>

<td> 327169</td><td> 693</td><td> 706</td><td> CACCTGGAAGCTGC</td><td> 3-8-3 MOE</td><td> 528</td>

<td> 327170</td><td> 694</td><td> 707</td><td> TCACCTGGAAGCTG</td><td> 3-8-3 MOE</td><td> 529</td>

<td> 327171</td><td> 695</td><td> 708</td><td> ATCACCTGGAAGCT</td><td> 3-8-3 MOE</td><td> 530</td>

<td> 327172</td><td> 696</td><td> 709</td><td> CATCACCTGGAAGC</td><td> 3-8-3 MOE</td><td> 531</td>

<td> 327173</td><td> 697</td><td> 710</td><td> ACATCACCTGGAAG</td><td> 3-8-3 MOE</td><td> 532</td>

<td> 327174</td><td> 698</td><td> 711</td><td> TACATCACCTGGAA</td><td> 3-8-3 MOE</td><td> 533</td>

<td> 327175</td><td> 699</td><td> 712</td><td> GTACATCACCTGGA</td><td> 3-8-3 MOE</td><td> 534</td>

<td> 327176</td><td> 700</td><td> 713</td><td> TGTACATCACCTGG</td><td> 3-8-3 MOE</td><td> 535</td>

<td> 327177</td><td> 701</td><td> 714</td><td> GTGTACATCACCTG</td><td> 3-8-3 MOE</td><td> 536</td>

<td> 327178</td><td> 869</td><td> 882</td><td> TAGCGGGTCCTGAG</td><td> 3-8-3 MOE</td><td> 537</td>

<td> 327179</td><td> 870</td><td> 883</td><td> GTAGCGGGTCCTGA</td><td> 3-8-3 MOE</td><td> 538</td>

<td> 327180</td><td> 871</td><td> 884</td><td> TGTAGCGGGTCCTG</td><td> 3-8-3 MOE</td><td> 539</td>

<td> 327181</td><td> 872</td><td> 885</td><td> CTGTAGCGGGTCCT</td><td> 3-8-3 MOE</td><td> 540</td>

<td> 327182</td><td> 873</td><td> 886</td><td> GCTGTAGCGGGTCC</td><td> 3-8-3 MOE</td><td> 541</td>

<td> 327183</td><td> 874</td><td> 887</td><td> GGCTGTAGCGGGTC</td><td> 3-8-3 MOE</td><td> 542</td>

<td> 327184</td><td> 875</td><td> 888</td><td> TGGCTGTAGCGGGT</td><td> 3-8-3 MOE</td><td> 543</td>

<td> 327185</td><td> 876</td><td> 889</td><td> CTGGCTGTAGCGGG</td><td> 3-8-3 MOE</td><td> 544</td>

<td> 327186</td><td> 877</td><td> 890</td><td> TCTGGCTGTAGCGG</td><td> 3-8-3 MOE</td><td> 545</td>

<td> 327187</td><td> 878</td><td> 891</td><td> TTCTGGCTGTAGCG</td><td> 3-8-3 MOE</td><td> 546</td>

<td> 327188</td><td> 955</td><td> 968</td><td> TGAACACCGCGGCC</td><td> 3-8-3 MOE</td><td> 547</td>

<td> 327189</td><td> 956</td><td> 969</td><td> ATGAACACCGCGGC</td><td> 3-8-3 MOE</td><td> 548</td>

<td> 327190</td><td> 957</td><td> 970</td><td> CATGAACACCGCGG</td><td> 3-8-3 MOE</td><td> 549</td>

<td> 327191</td><td> 958</td><td> 971</td><td> GCATGAACACCGCG</td><td> 3-8-3 MOE</td><td> 550</td>

<td> 327192</td><td> 959</td><td> 972</td><td> TGCATGAACACCGC</td><td> 3-8-3 MOE</td><td> 551</td>

<td> 327193</td><td> 960</td><td> 973</td><td> TTGCATGAACACCG</td><td> 3-8-3 MOE</td><td> 552</td>

<td> 327194</td><td> 961</td><td> 974</td><td> ATTGCATGAACACC</td><td> 3-8-3 MOE</td><td> 553</td>

<td> 327195</td><td> 962</td><td> 975</td><td> TATTGCATGAACAC</td><td> 3-8-3 MOE</td><td> 554</td>

<td> 327196</td><td> 963</td><td> 976</td><td> ATATTGCATGAACA</td><td> 3-8-3 MOE</td><td> 555</td>

<td> 327197</td><td> 964</td><td> 977</td><td> CATATTGCATGAAC</td><td> 3-8-3 MOE</td><td> 556</td>

<td> 327198</td><td> 1019</td><td> 1032</td><td> AGGTTGTGCAGGTA</td><td> 3-8-3 MOE</td><td> 557</td>

<td> 327199</td><td> 1020</td><td> 1033</td><td> CAGGTTGTGCAGGT</td><td> 3-8-3 MOE</td><td> 558</td>

<td> 327200</td><td> 1021</td><td> 1034</td><td> GCAGGTTGTGCAGG</td><td> 3-8-3 MOE</td><td> 559</td>

<td> 327201</td><td> 1022</td><td> 1035</td><td> AGCAGGTTGTGCAG</td><td> 3-8-3 MOE</td><td> 560</td>

<td> 327202</td><td> 1023</td><td> 1036</td><td> CAGCAGGTTGTGCA</td><td> 3-8-3 MOE</td><td> 561</td>

<td> 327203</td><td> 1024</td><td> 1037</td><td> CCAGCAGGTTGTGC</td><td> 3-8-3 MOE</td><td> 562</td>

<td> 327204</td><td> 1025</td><td> 1038</td><td> CCCAGCAGGTTGTG</td><td> 3-8-3 MOE</td><td> 563</td>

<td> 327205</td><td> 1026</td><td> 1039</td><td> GCCCAGCAGGTTGT</td><td> 3-8-3 MOE</td><td> 564</td>

<td> 327206</td><td> 1027</td><td> 1040</td><td> GGCCCAGCAGGTTG</td><td> 3-8-3 MOE</td><td> 565</td>

<td> 327207</td><td> 1028</td><td> 1041</td><td> AGGCCCAGCAGGTT</td><td> 3-8-3 MOE</td><td> 566</td>

<td> 338491</td><td> 1160</td><td> 1175</td><td> TGTCATTGCTGGTCCA</td><td> 3-10-3 MOE</td><td> 567</td>

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<td> 338562</td><td> 1160</td><td> 1173</td><td> TCATTGCTGGTCCA</td><td> 3-8-3 MOE</td><td> 568</td>

<td> 338498</td><td> 1307</td><td> 1322</td><td> TGGCCAGCCGGAACTT</td><td> 3-10-3 MOE</td><td> 569</td>

<td> 338569</td><td> 1307</td><td> 1320</td><td> GCCAGCCGGAACTT</td><td> 3-8-3 MOE</td><td> 570</td>

<td> 338499</td><td> 1329</td><td> 1344</td><td> GGGATGAGGGTCAGCG</td><td> 3-10-3 MOE</td><td> 571</td>

<td> 338570</td><td> 1329</td><td> 1342</td><td> GATGAGGGTCAGCG</td><td> 3-8-3 MOE</td><td> 572</td>

<td> 385067</td><td> 1364</td><td> 1377</td><td> AAGGCAAAGACCAC</td><td> 3-8-3 MOE</td><td> 573</td>

<td> 338573</td><td> 1401</td><td> 1414</td><td> GGAGCGCAGGGTGC</td><td> 3-8-3 MOE</td><td> 574</td>

<td> 338580</td><td> 1487</td><td> 1500</td><td> TGCACCTCCTTGTT</td><td> 3-8-3 MOE</td><td> 575</td>

Table 13: Short antisense compounds targeted to SEQ ID NO: I and having 1 or 2 mismatches

<td> ISIS NO.</td><td> 5’ Target Site</td><td> 3’ Target Site</td><td> Sequence (5’-3’)</td><td> Gapmer Motif</td><td> SEQ ID NO</td>

<td> 338577</td><td> 158</td><td> 171</td><td> CAGCAGACCCTGGA</td><td> 3-8-3 MOE</td><td> 576</td>

<td> 338458</td><td> 237</td><td> 252</td><td> ACATCTGGCAGAGGTT</td><td> 3-10-3 MOE</td><td> 577</td>

<td> 338529</td><td> 237</td><td> 250</td><td> ATCTGGCAGAGGTT</td><td> 3-8-3 MOE</td><td> 578</td>

<td> 338466</td><td> 318</td><td> 333</td><td> CAGGCCAGCAGGAGTA</td><td> 3-10-3 MOE</td><td> 579</td>

<td> 338537</td><td> 318</td><td> 331</td><td> GGCCAGCAGGAGTA</td><td> 3-8-3 MOE</td><td> 580</td>

<td> 338533</td><td> 364</td><td> 377</td><td> CAAACAAAAAGTCC</td><td> 3-8-3 MOE</td><td> 582</td>

<td> 338462</td><td> 364</td><td> 379</td><td> CTCAAACAAAAAGTCC</td><td> 3-10-3 MOE</td><td> 581</td>

<td> 338535</td><td> 397</td><td> 410</td><td> GGTGACATTGGTCA</td><td> 3-8-3 MOE</td><td> 584</td>

<td> 338464</td><td> 397</td><td> 412</td><td> GTGGTGACATTGGTCA</td><td> 3-10-3 MOE</td><td> 583</td>

<td> 338466</td><td> 470</td><td> 485</td><td> CAGGCCAGCAGGAGTA</td><td> 3-10-3 MOE</td><td> 579</td>

<td> 338537</td><td> 470</td><td> 483</td><td> GGCCAGCAGGAGTA</td><td> 3-8-3 MOE</td><td> 580</td>

<td> 385048</td><td> 497</td><td> 510</td><td> TTGGCAGTGGTGTT</td><td> 3-8-3 MOE</td><td> 587</td>

<td> 385049</td><td> 500</td><td> 513</td><td> ATGTTGGCAGTGGT</td><td> 3-8-3 MOE</td><td> 588</td>

<td> 338467</td><td> 503</td><td> 518</td><td> AGGAAATGTTGGCAGT</td><td> 3-10-3 MOE</td><td> 589</td>

<td> 338538</td><td> 503</td><td> 516</td><td> GAAATGTTGGCAGT</td><td> 3-8-3 MOE</td><td> 590</td>

<td> 385050</td><td> 506</td><td> 519</td><td> CAGGAAATGTTGGC</td><td> 3-8-3 MOE</td><td> 591</td>

<td> 385051</td><td> 509</td><td> 522</td><td> GGGCAGGAAATGTT</td><td> 3-8-3 MOE</td><td> 592</td>

<td> 385052</td><td> 523</td><td> 536</td><td> AAGGTAGGTACCAG</td><td> 3-8-3 MOE</td><td> 593</td>

<td> 385053</td><td> 526</td><td> 539</td><td> ACCAAGGTAGGTAC</td><td> 3-8-3 MOE</td><td> 594</td>

<td> 385056</td><td> 535</td><td> 548</td><td> CTTTGTGGCACCAA</td><td> 3-8-3 MOE</td><td> 595</td>

<td> 385057</td><td> 538</td><td> 551</td><td> GCACTTTGTGGCAC</td><td> 3-8-3 MOE</td><td> 596</td>

<td> 338539</td><td> 539</td><td> 552</td><td> TGCACTTTGTGGCA</td><td> 3-8-3 MOE</td><td> 597</td>

<td> 385058</td><td> 541</td><td> 554</td><td> GCTGCACTTTGTGG</td><td> 3-8-3 MOE</td><td> 598</td>

<td> 385059</td><td> 544</td><td> 557</td><td> GGTGCTGCACTTTG</td><td> 3-8-3 MOE</td><td> 599</td>

<td> 385060</td><td> 547</td><td> 560</td><td> GGCGGTGCTGCACT</td><td> 3-8-3 MOE</td><td> 600</td>

<td> 385063</td><td> 556</td><td> 569</td><td> TGAACACTAGGCGG</td><td> 3-8-3 MOE</td><td> 601</td>

<td> 385064</td><td> 559</td><td> 572</td><td> TCTTGAACACTAGG</td><td> 3-8-3 MOE</td><td> 602</td>

<td> 338469</td><td> 561</td><td> 576</td><td> CACCTCTTGAACACTA</td><td> 3-10-3 MOE</td><td> 603</td>

<td> 338540</td><td> 561</td><td> 574</td><td> CCTCTTGAACACTA</td><td> 3-8-3 MOE</td><td> 604</td>

<td> 385065</td><td> 562</td><td> 575</td><td> ACCTCTTGAACACT</td><td> 3-8-3 MOE</td><td> 605</td>

<td> 385066</td><td> 565</td><td> 578</td><td> CACACCTCTTGAAC</td><td> 3-8-3 MOE</td><td> 606</td>

<td> 338541</td><td> 590</td><td> 603</td><td> CCTCGAACCCACTG</td><td> 3-8-3 MOE</td><td> 607</td>

<td> 338473</td><td> 658</td><td> 673</td><td> CTTCTGGACCTCGATC</td><td> 3-10-3 MOE</td><td> 608</td>

<td> 338544</td><td> 658</td><td> 671</td><td> TCTGGACCTCGATC</td><td> 3-8-3 MOE</td><td> 609</td>

<td> 338474</td><td> 681</td><td> 696</td><td> CTGCTATACATCTTGG</td><td> 3-10-3 MOE</td><td> 610</td>

<td> 338545</td><td> 681</td><td> 694</td><td> GCTATACATCTTGG</td><td> 3-8-3 MOE</td><td> 611</td>

<td> 338475</td><td> 703</td><td> 718</td><td> CACGGTGTACATCACC</td><td> 3-10-3 MOE</td><td> 612</td>

<td> 338546</td><td> 703</td><td> 716</td><td> CGGTGTACATCACC</td><td> 3-8-3 MOE</td><td> 613</td>

<td> 338547</td><td> 718</td><td> 731</td><td> ACAGACTGTAGCCC</td><td> 3-8-3 MOE</td><td> 615</td>

<td> 338476</td><td> 718</td><td> 733</td><td> GGACAGACTGTAGCCC</td><td> 3-10-3 MOE</td><td> 614</td>

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<td> 338550</td><td> 889</td><td> 902</td><td> CATCGCCAATCTTC</td><td> 3-8-3 MOE</td><td> 617</td>

<td> 338479</td><td> 889</td><td> 904</td><td> GTCATCGCCAATCTTC</td><td> 3-10-3 MOE</td><td> 616</td>

<td> 338551</td><td> 899</td><td> 912</td><td> ACACTGAGGTCATC</td><td> 3-8-3 MOE</td><td> 619</td>

<td> 338480</td><td> 899</td><td> 914</td><td> TCACACTGAGGTCATC</td><td> 3-10-3 MOE</td><td> 618</td>

<td> 338552</td><td> 924</td><td> 937</td><td> CGCCCCGTCACTGA</td><td> 3-8-3 MOE</td><td> 620</td>

<td> 338555</td><td> 992</td><td> 1005</td><td> AGCAACCAGCAATA</td><td> 3-8-3 MOE</td><td> 622</td>

<td> 338484</td><td> 992</td><td> 1007</td><td> CCAGCAACCAGCAATA</td><td> 3-10-3 MOE</td><td> 621</td>

<td> 338485</td><td> 1018</td><td> 1033</td><td> CAGGCTGTACAGGTAC</td><td> 3-10-3 MOE</td><td> 623</td>

<td> 338556</td><td> 1018</td><td> 1031</td><td> GGCTGTACAGGTAC</td><td> 3-8-3 MOE</td><td> 624</td>

<td> 338558</td><td> 1051</td><td> 1064</td><td> AGCTCCTCTCAGAG</td><td> 3-8-3 MOE</td><td> 626</td>

<td> 338487</td><td> 1051</td><td> 1066</td><td> GAAGCTCCTCTCAGAG</td><td> 3-10-3 MOE</td><td> 625</td>

<td> 338559</td><td> 1079</td><td> 1092</td><td> CAGCCAATGCCCAG</td><td> 3-8-3 MOE</td><td> 628</td>

<td> 338488</td><td> 1079</td><td> 1094</td><td> CCCAGCCAATGCCCAG</td><td> 3-10-3 MOE</td><td> 627</td>

<td> 338560</td><td> 1131</td><td> 1144</td><td> AAACAGACACTTGA</td><td> 3-8-3 MOE</td><td> 630</td>

<td> 338489</td><td> 1131</td><td> 1146</td><td> TCAAACAGACACTTGA</td><td> 3-10-3 MOE</td><td> 629</td>

<td> 338490</td><td> 1145</td><td> 1160</td><td> AGCACTGAACATTCTC</td><td> 3-10-3 MOE</td><td> 631</td>

<td> 338561</td><td> 1145</td><td> 1158</td><td> CACTGAACATTCTC</td><td> 3-8-3 MOE</td><td> 632</td>

<td> 338563</td><td> 1181</td><td> 1194</td><td> ATCCACCAGAATCC</td><td> 3-8-3 MOE</td><td> 634</td>

<td> 338492</td><td> 1181</td><td> 1196</td><td> GGATCCACCAGAATCC</td><td> 3-10-3 MOE</td><td> 633</td>

<td> 338564</td><td> 1216</td><td> 1229</td><td> TGATCAGTAAGGCC</td><td> 3-8-3 MOE</td><td> 635</td>

<td> 338565</td><td> 1232</td><td> 1245</td><td> ACAAAGATGAAAAA</td><td> 3-8-3 MOE</td><td> 637</td>

<td> 338494</td><td> 1232</td><td> 1247</td><td> GGACAAAGATGAAAAA</td><td> 3-10-3 MOE</td><td> 636</td>

<td> 338566</td><td> 1267</td><td> 1280</td><td> CACGCAGCTTGGCC</td><td> 3-8-3 MOE</td><td> 639</td>

<td> 338495</td><td> 1267</td><td> 1282</td><td> GGCACGCAGCTTGGCC</td><td> 3-10-3 MOE</td><td> 638</td>

<td> 338571</td><td> 1344</td><td> 1357</td><td> GACCCCCAGCAGAG</td><td> 3-8-3 MOE</td><td> 641</td>

<td> 338500</td><td> 1344</td><td> 1359</td><td> TGGACCCCCAGCAGAG</td><td> 3-10-3 MOE</td><td> 640</td>

<td> 385068</td><td> 1366</td><td> 1379</td><td> CAAAGGCAAAGACC</td><td> 3-8-3 MOE</td><td> 642</td>

<td> 385069</td><td> 1369</td><td> 1382</td><td> TCACAAAGGCAAAG</td><td> 3-8-3 MOE</td><td> 643</td>

<td> 385070</td><td> 1372</td><td> 1385</td><td> CAGTCACAAAGGCA</td><td> 3-8-3 MOE</td><td> 644</td>

<td> 385071</td><td> 1375</td><td> 1388</td><td> CGTCAGTCACAAAG</td><td> 3-8-3 MOE</td><td> 645</td>

<td> 385072</td><td> 1378</td><td> 1391</td><td> GCTCGTCAGTCACA</td><td> 3-8-3 MOE</td><td> 646</td>

<td> 385073</td><td> 1381</td><td> 1394</td><td> CÀTGCTCGTCAGTC</td><td> 3-8-3 MOE</td><td> 647</td>

<td> 386608</td><td> 1384</td><td> 1397</td><td> GGGCATGCTCGTCA</td><td> 1-12-1 MOE</td><td> 648</td>

<td> 386593</td><td> 1384</td><td> 1397</td><td> GGGCATGCTCGTCA</td><td> 2-10-2 MOE</td><td> 648</td>

<td> 396146</td><td> 1384</td><td> 1397</td><td> GGGCATGCTCGTCA</td><td> 2-10-2 MOE</td><td> 648</td>

<td> 338572</td><td> 1384</td><td> 1397</td><td> GGGCATGCTCGTCA</td><td> 3-8-3 MOE</td><td> 648</td>

<td> 396149</td><td> 1384</td><td> 1397</td><td> GGGCATGCTCGTCA</td><td> 1-1-10-22’- (butylacetamido)palmitamide/OMe/ OMe</td><td> 648</td>

<td> 386627</td><td> 1384</td><td> 1397</td><td> GGGCATGCTCGTCA</td><td> 2-10-2 Methyleneoxy BNA</td><td> 648</td>

<td> 386610</td><td> 1387</td><td> 1400</td><td> CTTGGGCATGCTCG</td><td> 1-12-1 MOE</td><td> 654</td>

<td> 386595</td><td> 1387</td><td> 1400</td><td> CTTGGGCATGCTCG</td><td> 2-10-2 MOE</td><td> 654</td>

<td> 385074</td><td> 1387</td><td> 1400</td><td> CTTGGGCATGCTCG</td><td> 3-8-3 MOE</td><td> 654</td>

<td> 385075</td><td> 1390</td><td> 1403</td><td> TGCCTTGGGCATGC</td><td> 3-8-3 MOE</td><td> 657</td>

<td> 385076</td><td> 1393</td><td> 1406</td><td> GGGTGCCTTGGGCA</td><td> 3-8-3 MOE</td><td> 648</td>

<td> 385077</td><td> 1396</td><td> 1409</td><td> GCAGGGTGCCTTGG</td><td> 3-8-3 MOE</td><td> 659</td>

<td> 385078</td><td> 1399</td><td> 1412</td><td> AGCGCAGGGTGCCT</td><td> 3-8-3 MOE</td><td> 660</td>

<td> 338502</td><td> 1401</td><td> 1416</td><td> GTGGAGCGCAGGGTGC</td><td> 3-10-3 MOE</td><td> 661</td>

<td> 385079</td><td> 1402</td><td> 1415</td><td> TGGAGCGCAGGGTG</td><td> 3-8-3 MOE</td><td> 662</td>

<td> 385080</td><td> 1405</td><td> 1418</td><td> TGGTGGAGCGCAGG</td><td> 3-8-3 MOE</td><td> 663</td>

<td> 385081</td><td> 1408</td><td> 1421</td><td> GCTTGGTGGAGCGC</td><td> 3-8-3 MOE</td><td> 664</td>

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<td> 385082</td><td> 1411</td><td> 1424</td><td> AGAGCTTGGTGGAG</td><td> 3-8-3 MOE</td><td> 665</td>

<td> 338503</td><td> 1412</td><td> 1427</td><td> AAAAGAGCTTGGTGGA</td><td> 3-10-3 MOE</td><td> 666</td>

<td> 338574</td><td> 1412</td><td> 1425</td><td> AAGAGCTTGGTGGA</td><td> 3-8-3 MOE</td><td> 667</td>

<td> 385083</td><td> 1414</td><td> 1427</td><td> AAAAGAGCTTGGTG</td><td> 3-8-3 MOE</td><td> 668</td>

<td> 385084</td><td> 1417</td><td> 1430</td><td> CAAAAAAGAGCTTG</td><td> 3-8-3 MOE</td><td> 669</td>

<td> 338504</td><td> 1434</td><td> 1449</td><td> AAGGAGCTGAGGAACA</td><td> 3-10-3 MOE</td><td> 670</td>

<td> 338575</td><td> 1434</td><td> 1447</td><td> GGAGCTGAGGAACA</td><td> 3-8-3 MOE</td><td> 671</td>

<td> 327167</td><td> 1441</td><td> 1454</td><td> CCTGGAAGCTGCTG</td><td> 3-8-3 MOE</td><td> 526</td>

<td> 338576</td><td> 1445</td><td> 1458</td><td> AGACCCTGGAAGGA</td><td> 3-8-3 MOE</td><td> 673</td>

<td> 338505</td><td> 1445</td><td> 1460</td><td> GCAGACCCTGGAAGGA</td><td> 3-10-3 MOE</td><td> 672</td>

<td> 338506</td><td> 1449</td><td> 1464</td><td> ACCAGCAGACCCTGGA</td><td> 3-10-3 MOE</td><td> 674</td>

<td> 338577</td><td> 1449</td><td> 1462</td><td> CAGCAGACCCTGGA</td><td> 3-8-3 MOE</td><td> 576</td>

<td> 338507</td><td> 1464</td><td> 1479</td><td> CAGTAGAGAACAGCCA</td><td> 3-10-3 MOE</td><td> 676</td>

<td> 338578</td><td> 1464</td><td> 1477</td><td> GTAGAGAACAGCCA</td><td> 3-8-3 MOE</td><td> 677</td>

<td> 338508</td><td> 1475</td><td> 1490</td><td> TGTTGAGGAAACAGTA</td><td> 3-10-3 MOE</td><td> 678</td>

<td> 338579</td><td> 1475</td><td> 1488</td><td> TTGAGGAAACAGTA</td><td> 3-8-3 MOE</td><td> 679</td>

<td> 338509</td><td> 1487</td><td> 1502</td><td> CCTGCACCTCCTTGTT</td><td> 3-10-3 MOE</td><td> 680</td>

<td> 338580</td><td> 1610</td><td> 1623</td><td> TGCACCTCCTTGTT</td><td> 3-8-3 MOE</td><td> 575</td>

In certain embodiments, a target region is nucleotides 378-391 of SEQ ID NO: 9. In certain embodiments, a short antisense compound is targeted to nucleotides 378-391 of SEQ ID NO: 9. In certain such embodiments, a short antisense compound targeted to nucleotides 378-391 comprises a nucleotide sequence selected from SEQ ID NO 486 or 487. In certain such embodiments, a short antisense compound targeted to nucleotides 378-391 of SEQ ID NO: 9 is selected from Isis No 338463 or 338534.

In certain embodiments, a target region is nucleotides 499-521 of SEQ ID NO; 9. In certain embodiments, a short antisense compound is targeted to nucleotides 499-521 of SEQ ID NO: 9. In certain such embodiments, a short antisense compound targeted to nucleotides 499-521 comprises a nucleotide sequence selected from SEQ ID NO 488,489,490,491,492,493,494,495,496, or 497. In certain such embodiments, a short antisense compound targeted to nucleotides 499-521 of SEQ ID NO: 9 is selected from Isis No 327130,327131,327132,327133,327134,327135,327136,327137,327138, or 327139.

In certain embodiments, a target region is nucleotides 531-553 of SEQ ID NO: 9. In certain embodiments, a short antisense compound is targeted to nucleotides 531-553 of SEQ ID NO: 9. In certain such embodiments, a short antisense compound targeted to nucleotides 531-553 comprises a nucleotide sequence selected from SEQ ID NO 498,499,500, 501, 502,503,504,505,506, or 507. In certain such embodiments, a short antisense compound targeted to nucleotides 531-553 of SEQ ID NO: 9 is selected from Isis No 327140,327141,327142,327143,327144,327145,327146,327147,327148, or 327149.

In certain embodiments, a target region is nucleotides 545-567 of SEQ ID NO: 9. In certain embodiments, a short antisense compound is targeted to nucleotides 545-567 of SEQ ID NO: 9. In certain such embodiments, a short antisense compound targeted to nucleotides 545-567 comprises a nucleotide sequence selected from SEQ ID NO 508,509,510, 511,512,513,514,515,516, or 517. In certain such embodiments, a short antisense compound targeted to nucleotides 545-567 of SEQ ID NO: 9 is selected from Isis No 327150,327151,327152,327153,327154,327155,327156,327157,327158, or 327159.

In certain embodiments, a target region is nucleotides 531-567 of SEQ ID NO: 9. In certain

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-124embodiments, a short antisense compound is targeted to nucleotides 531-567 of SEQ ID NO: 9. In certain such embodiments, a short antisense compound targeted to nucleotides 531-567 comprises a nucleotide sequence selected from SEQ ID NO 498,499,500,501,502,503,504,505,506,507,508,509,510,511, 512, 513, 514, 515, 516, or 517. In certain such embodiments, a short antisense compound targeted to nucleotides 531-567 of SEQ ID NO: 9 is selected from Isis No 327140, 327141, 327142, 327143, 327144, 327145, 327146, 327147, 327148, 327149, 327150, 327151, 327152, 327153, 327154, 327155, 327156,327157,327158, or 327159.

In certain embodiments, a target region is nucleotides 684-714 of SEQ ID NO: 9. In certain embodiments, a short antisense compound is targeted to nucleotides 684-714 of SEQ ID NO: 9. In certain such embodiments, a short antisense compound targeted to nucleotides 684-714 comprises a nucleotide sequence selected from SEQ ID NO 518,520,521,522,523,524,525,526,527,528,529,530,531,532, 533, 534, 535, or 536. In certain such embodiments, a short antisense compound targeted to nucleotides 684-714 of SEQ ID NO: 9 is selected from Isis No 345897,327160,327161, 327162, 327163, 327164, 327165, 327166, 327167, 327168, 327169, 327170, 327171, 327172,327173, 327174, 327175, 327176, or 327177.

In certain embodiments, a target region is nucleotides 869-891 of SEQ ID NO: 9. In certain embodiments, a short antisense compound is targeted to nucleotides 869-891 of SEQ ID NO: 9. In certain such embodiments, a short antisense compound targeted to nucleotides 869-891 comprises a nucleotide sequence selected from SEQ ID NO 537, 538,539,540,541,542,543,544,545, or 546. In certain such embodiments, a short antisense compound targeted to nucleotides 869-891 of SEQ ID NO; 9 is selected from Isis No 327178,327179,327180,327181,327182,327183,327184,327185,327186, or 327187.

In certain embodiments, a target region is nucleotides 955-977 of SEQ ID NO: 9. In certain embodiments, a short antisense compound is targeted to nucleotides 955-977 of SEQ ID NO: 9. In certain such embodiments, a short antisense compound targeted to nucleotides 955-977 comprises a nucleotide sequence selected from SEQ ID NO 547,548,549, 550,551,552,553, 554,555, or 556. In certain such embodiments, a short antisense compound targeted to nucleotides 955-977 of SEQ ID NO: 9 is selected from Isis No 327188,327189,327190,327191,327192,327193,327194,327195,327196, or 327197.

In certain embodiments, a target region is nucleotides 1019-1041 of SEQ ID NO: 9. In certain embodiments, a short antisense compound is targeted to nucleotides 1019-1041 of SEQ ID NO: 9. In certain such embodiments, a short antisense compound targeted to nucleotides 1019-1041 comprises a nucleotide sequence selected from SEQ ID NO 557, 558, 559, 560, 561, 562, 563, 564, 565, or 566. In certain such embodiments, a short antisense compound targeted to nucleotides 1019-1041 of SEQ ID NO: 9 is selected from Isis No 327198,327199,327200,327201,327202, 327203,327204, 327205, 327206, or 327207.

In certain embodiments, a target region is nucleotides 1160-1175 of SEQ ID NO: 9. In certain embodiments, a short antisense compound is targeted to nucleotides 1160-1175 of SEQ ID NO: 9. In certain such embodiments, a short antisense compound targeted to nucleotides 1160-1175 comprises a

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-125nucleotide sequence selected from SEQ ID NO 567 or 568. In certain such embodiments, a short antisense compound targeted to nucleotides 1160-1175 of SEQ ID NO: 9 is selected from Isis No 338491 or 338562.

In certain embodiments, a target region is nucleotides 1307-1377 of SEQ ID NO: 9. In certain embodiments, a short antisense compound is targeted to nucleotides 1307-1377 of SEQ ID NO: 9. In certain such embodiments, a short antisense compound targeted to nucleotides 1307-1377 comprises a nucleotide sequence selected from SEQ ID NO 569,570,571,572, or 573. In certain such embodiments, a short antisense compound targeted to nucleotides 1307-1377 of SEQ ID NO: 9 is selected from Isis No 338498,338569,338499,338570, or 385067.

In certain embodiments, a target region is nucleotides 1307-1414 of SEQ ID NO: 9. In certain embodiments, a short antisense compound is targeted to nucleotides 1307-1414 of SEQ ID NO: 9. In certain such embodiments, a short antisense compound targeted to nucleotides 1307-1414 comprises a nucleotide sequence selected from SEQ ID NO 569, 570, 571, 572, 573, or 574. In certain such embodiments, a short antisense compound targeted to nucleotides 1307-1414 of SEQ ID NO: 9 is selected from Isis No 338498,338569,338499,338570,385067, or 338573.

In certain embodiments, short antisense compounds targeted to a GCGR nucleic acid are 8 to 16, preferably 9 to 15, more preferably 9 to 14, more preferably 10 to 14 nucleotides in length. In certain embodiments, short antisense compounds targeted to a GCGR nucleic acid are 9 to 14 nucleotides in length. In certain embodiments, short antisense compounds targeted to a GCGR nucleic acid are 10 to 14 nucleotides in length. In certain embodiments, such short antisense compounds are short antisense oligonucleotides.

In certain embodiments, short antisense compounds targeted to a GCGR nucleic acid are short gapmers. In certain such embodiments, short gapmers targeted to a GCGR nucleic acid comprise at least one high affinity modification in one or more wings of the compound. In certain embodiments, short antisense compounds targeted to a GCGR nucleic acid comprise 1 to 3 high-affinity modifications in each wing. In certain such embodiments, the nucleosides or nucleotides of the wing comprise a 2’ modification. In certain such embodiments, the monomers of the wing are BNA’s. In certain such embodiments, the monomers of the wing are selected from α-L-Methyleneoxy (4’-CH₂-O-2’) BNA, β-DMethyleneoxy (4’-CH₂-O-2’) BNA , Ethyleneoxy (4’-(CH2)2-O-2’) BNA , Aminooxy (4’-CH₂-O-N(R)2’) BNA and Oxyamino (4’-CH2-N(R)-O-2’) BNA. In certain embodiments, the monomers of a wing comprise a substituent at the 2’ position selected from allyl, amino, azido, thio, O-allyl, O-C_rC₁₀ alkyl, OCFj, O-(CH2)₂-O-CH₃, 2'-O(CH₂)₂SCHj,O-(CH2)2-O-N(R_mXK₁), and O-CH₂-C(=O)-N(R_a)(R_n), where each R_a and R^ is, independently, H or substituted or unsubstituted C|-Cio alkyl. In certain embodiments, the monomers of a wing are 2’MOE nucleotides.

In certain embodiments, short antisense compounds targeted to a GCGR nucleic acid comprise a gap between the 5’ wing and the 3’ wing. In certain embodiments the gap comprises five, six, seven, eight, nine, ten, eleven, twelve, thirteen, or fourteen monomers. In certain embodiments, the monomers

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-126of the gap are unmodified deoxyribonucleotides. In certain embodiments, the monomers of the gap are unmodified ribonucleotides. In certain embodiments, gap modifications (if any) gap result in an antisense compound that, when bound to its target nucleic acid, supports cleavage by an RNase, including, but not limited to, RNase H.

In certain embodiments, short antisense compounds targeted to a GCGR nucleic acid have uniform monomeric linkages. In certain such embodiments, those linkages are all phosphorothioate linkages. In certain embodiments, the linkages are all phosphodiester linkages. In certain embodiments, short antisense compounds targeted to a GCGR nucleic acid have mixed backbones.

In certain embodiments, short antisense compounds targeted to a GCGR nucleic acid arc 8 monomers in length. In certain embodiments, short antisense compounds targeted to a GCGR nucleic acid are 9 monomers in length. In certain embodiments, short antisense compounds targeted to a GCGR nucleic acid are 10 monomers in length. In certain embodiments, short antisense compounds targeted to a GCGR nucleic acid are 11 monomers in length. In certain embodiments, short antisense compounds targeted to a GCGR nucleic acid are monomers in length. In certain embodiments, short antisense compounds targeted to a GCGR nucleic acid are 13 monomers in length. In certain embodiments, short antisense compounds targeted to a GCGR nucleic acid are 14 monomers in length. In certain embodiments, short antisense compounds targeted to a GCGR nucleic acid are 15 monomers in length. In certain embodiments, short antisense compounds targeted to a GCGR nucleic acid are 16 monomers in length. In certain embodiments, short antisense compounds targeted to a GCGR nucleic acid comprise 9 to 15 monomers. In certain embodiments, short antisense compounds targeted to a GCGR nucleic acid comprise 10 to 15 monomers. In certain embodiments, short antisense compounds targeted to a GCGR nucleic acid comprise 12 to 14 monomers. In certain embodiments, short antisense compounds targeted to a GCGR nucleic acid comprise 12 to 14 nucleotides or nucleosides.

In certain embodiments, the invention provides methods of modulating expression of GCGR. In certain embodiments, such methods comprise use of one or more short antisense compound targeted to a GCGR nucleic acid, wherein the short antisense compound targeted to a GCGR nucleic acid is from about 8 to about 16, preferably 9 to 15, more preferably 9 to 14, more preferably 10 to 14 monomers (i.e. from about 8 to about 16 linked monomers). One of ordinary skill in the art will appreciate that this comprehends methods of modulating expression of GCGR using one or more short antisense compounds targeted to a GCGR nucleic acid of 8,9,10,11,12,13,14,15 or 16monomers.

In certain embodiments, methods of modulating GCGR comprise use of a short antisense compound targeted to a GCGR nucleic acid that is 8 monomers in length. In certain embodiments, methods of modulating GCGR comprise use of a short antisense compound targeted to a GCGR nucleic acid frat is 9 monomers in length. In certain embodiments, methods of modulating GCGR comprise use of a short antisense compound targeted to a GCGR nucleic acid that is 10 monomers in length. In certain embodiments, methods of modulating GCGR comprise use of a short antisense compound targeted to a GCGR nucleic acid that is 11 monomers in length. In certain embodiments, methods of modulating

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-127GCGR comprise use of a short antisense compound targeted to a GCGR nucleic acid that is 12 monomers in length. In certain embodiments, methods of modulating GCGR comprise use of a short antisense compound targeted to a GCGR nucleic acid that is 13 monomers in length. In certain embodiments, methods of modulating GCGR comprise use of a short antisense compound targeted to a GCGR nucleic acid that is 14 monomers in length. In certain embodiments, methods of modulating GCGR comprise use of a short antisense compound targeted to a GCGR nucleic acid that is 15 monomers in length. In certain embodiments, methods of modulating GCGR comprise use of a short antisense compound targeted to a GCGR nucleic acid that is 16 monomers in length.

In certain embodiments, methods of modulating expression of GCGR comprise use of a short antisense compound targeted to a GCGR nucleic acid comprising 9 to 15 monomers. In certain embodiments, methods of modulating expression of GCGR comprise use of a short antisense compound targeted to a GCGR nucleic acid comprising 10 to 15 monomers. In certain embodiments, methods of modulating expression of GCGR comprise use of a short antisense compound targeted to a GCGR nucleic acid comprising 12 to 14 monomers. In certain embodiments, methods of modulating expression of GCGR comprise use of a short antisense compound targeted to a GCGR nucleic acid comprising 12 or 14 nucleotides or nucleosides.

8. DGAT2

Diacylglycerol transferase 2 (also known as DGAT2, diacylglycerol O-transferase 2, acylCoA:diacylglycerol acyltransferase 2), Diacylglycerol transferase 2 has been shown to be implicated in the absorption process of triglycerides (also called triacylglycerols) from food.

The absorption of triglycerides from food is a very efficient process which occurs by a series of steps wherein die dietary triacylglycerols are hydrolyzed in the intestinal lumen and then resynthesized within enterocytes. The resynthesis of triacylglycerols can occur via the monoacylglycerol pathwaywhich commences with monoacylglycerol acyltransferase (MGAT) catalyzing the synthesis of diacylglycerol from monoacylglycerol and fatty acyl-CoA. An alternative synthesis of diacylglycerols is provided by the glycerol-phosphate pathway which describes the coupling of two molecules of fatty acyl-CoA to glycerol-3-phosphate. In either case, diacylglycerol is then acylated with another molecule of fatty acylCoA in a reaction catalyzed by one of two diacylglycerol acyltransferase enzymes to form the triglyceride (Farese et al., Curr. Opin. Lipidol., 2000,11,229-234).

The reaction catalyzed by diacylglycerol acyltransferase is the final and only committed step in triglyceride synthesis. As such, diacylglycerol acyltransferase is involved in intestinal fat absorption, lipoprotein assembly, regulating plasma triglyceride concentrations, and fat storage in adipocytes. The first diacylglycerol acyltransferase, diacylglycerol transferase 1, was identified in 1960 and the human and mouse genes encoding this protein were isolated in 1998 (Cases et al., Proc. Nall. Acad. Sci. U. S. A., 1998, 95, 13018-13023; Oelkers et al., J. Biol. Chem., 1998, 273, 26765-26771). Mice lacking diacylglycerol acyltransferase 1 are viable and can still synthesize triglycerides through other biological

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-128routes, suggesting the existence of multiple mechanisms· for triglyceride synthesis (Smith et al., Nat. Gene/.. 2000,25,87-90).

A second diacylglycerol transferase, diacylglycerol transferase 2 (also known as DGAT2, diacylglycerol O-transferase 2, acyl-CoArdiacylglycerol acyltransferase 2), was subsequently identified in the fungus Mortierella, humans and mice (Cases et al., J. Biol. Chem., 2001, 276, 38870-38876; Lardizabal et al., J. Biol. Chem., 2001,276, 38862-38869). Enzymatic assays indicate that this recently identified protein does possess diacylglycerol transferase activity that utilizes a broad range of long chain fatty acyl-CoA substrates (Cases et al., J. Biol. Chem., 2001,276,38870-38876).

Diacylglycerol transferase 2 is a member of a family of genes whose sequences are unrelated to diacylglycerol transferase 1. In addition to differing in sequence compared to diacylglycerol transferase 1, in vitro assays illustrate that diacylglycerol transferase 2 has higher activity at lower concentrations of magnesium chloride and oleoyl-CoA (Cases et al., J. Biol. Chem., 2001, 276, 38870-38876). The predicted protein sequence of diacylglycerol transferase 2 contains at least one putative transmembrane domain, three potential N-linked glycosylation sites, six potential protein kinase C phosphorylation consensus sites, as well as sequences in common with a putative glycerol phosphorylation site found in acyltransferase enzymes (Cases et al., J. Biol. Chem., 2001, 276, 38870-38876). The International Radiation Hybrid Mapping Consortium has mapped human diacylglycerol transferase 2 to chromosome Uql3.3.

In human tissues, the highest levels of diacylglycerol transferase 2 are detected in liver and white adipose tissues, with lower levels found in mammary gland, testis and peripheral blood leukocytes (Cases et al., J. Biol. Chem., 2001,276,38870-38876). Two mRNA species of 2.4 and 1.8 kilobases are detected in human tissues, whereas the major diacylglycerol transferase 2 mRNA species in mouse tissues is 2.4 kilobases. In addition to liver and white adipose tissues, diacylglycerol transferase 2 is expressed in all segments of the small intestine in mice, with higher expression in the proximal intestine and lower expression in the distal intestine (Cases et al., J. Biol. Chem., 2001,276,38870-38876).

Diacylglycerol transferase activity exhibits distinct patterns during postnatal development of the rat liver. As there is no correlation between the mRNA expression and activity patterns, post-translational modifications may participate in the regulation of diacylglycerol transferase 2 activity during rat development (Waterman et al., J. Lipid. Res., 2002,43,1555-1562).

Diacylglycerol transferase 2 mRNA is preferentially upregulated by insulin treatment, as shown by in vitro assays measuring the diacylglycérol activity from the membrane fraction of cultured mouse adipocytes (Meegalla et al., Biochem. Biophys. Res. Commun., 2002, 298, 317-323). In fasting mice, diacylglycérol transferase 2 expression is greatly reduced, and dramatically increases upon refeeding. The expression patterns of two enzymes that participate in fatty acid synthesis, acetyl-CoA carboxylase and fatty acid synthase, respond to fasting and refeeding in a similar fashion. These results, combined with the observation that diacylglycerol transferase 2 is abundantly expressed in liver, suggest that diacylglycerol transferase 2 is tightly linked to the endogenous fatty acid synthesis pathway (Meegalla et

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-129al., Biochem. Biophys. Res. Commun., 2002,298,317-323).

Studies of mice harboring a disruption in the diacylglycerol acyltransferase 1 gene provide evidence that diacylglycerol acyltransferase 2 contributes to triglyceride synthesis. Levels of diacylglycerol transferase 2 mRNA expression are similar in intestinal segments from both wild type and diacylglycerol transferase 1-deficient mice (Buhman et al., J. Biol. Chem., 2002, 277, 25474-25479). Using magnesium chloride to distinguish between diacylglycérol transferase 1 and 2 activity, Buhman, et al. observed that, in diacylglycerol transferase 1-deficient mice, diacylglycerol transferase activity is reduced to 50% in the proximal intestine and to 10-15% in the distal intestine (Buhman et al., J. Biol. Chem., 2002,277,25474-25479).

Additionally, diacylglycerol transferase 2 mRNA levels are not up-regulated the liver or adipose tissues of diacylglycerol transferase 1-deficient mice, even after weeks of high-fat diet (Cases et ai., J. Biol. Chem., 2001,276, 38870-38876; Chen et al., J. Clin. Invest., 2002,109, 1049-1055). However, in ob/ob mice, which have a mutation in the leptin gene that results in obesity, diacylglycerol transferase 2 is more highly expressed than in wild type mice, suggesting that diacylglycérol transferase 2 may be partly responsible for the highly accumulated fat mass seen in these mice. Furthermore, the combined mutations of leptin and diacylglycerol transferase 1 leads to a three-fold elevation in diacylglycerol transferase 2 expression in white adipose tissue, compared to the levels in the same tissue from diacylglycerol transferase 1-deficient mice (Chen et al., J. Clin. Invest., lOGil, 109, 1049-1055). Diacylglycerol transferase 2 mRNA is also upregulated in the skin of these mice (Chen et al., J. Clin. Invest., 2002,109, 175-181). These data suggest leptin normally downregulates diacylglycerol transferase 2 expression, and that the upregulation of diacylglycerol transferase 2 in white adipose tissue in these mice may provide an alternate pathway for the triglyceride synthesis that still occurs in leptin deficient/ diacylglycerol transferase 1-deficient mice (Chen et al., J. Clin. Invest., 2002,109,1049-1055).

Diacylglycerol acyltransferase 1 knockout mice exhibit interesting phenotypes in that they are lean, resistant to diet-induce obesity, have decreased levels of tissue triglycerides and increased sensitivity to insulin and leptin (Chen et al., J. Clin. Invest., 2002,109, 1049-1055; Smith et al., Nat. Genet., 2000, 25, 87-90). As diacylglycerol transferase 2 also participates in triglyceride synthesis, interfering with diacylglycerol transferase 2 may similarly lead to reduced body fat content.

Definitions “DGAT2” means the gene product or protein of which expression is to be modulated by administration of a short antisense compound.

“DGAT2 nucleic acid means any nucleic acid encoding DGAT2. For example, in certain embodiments, a DGAT2 nucleic acid includes, without limitation, a DNA sequence encoding DGAT2, an RNA sequence transcribed from DNA encoding DGAT2, and an mRNA sequence encoding DGAT2.

“DGAT2 mRNA” means an mRNA encoding DGAT2.

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-130Therapeulic indications

Antisense technology is an effective means for reducing DGAT2 expression and has proven to be uniquely useful in a number of therapeutic, diagnostic, and research applications. As such, in certain embodiments, the present invention provides compounds targeted to nucleic acid encoding DGAT2, which modulate the expression of DGAT2. Further provided herein are short antisense compounds capable of effectively inhibiting DGAT2 expression.

In certain embodiments, a subject, suspected of having a disease or associated with DGAT2 is treated by administering one or more short antisense compounds targeted to a nucleic acid encoding DGAT2. For example, in a non-limiting embodiment, such methods comprise the step of administering to an animal a therapeutically effective amount of a short antisense compound. In certain such embodiments, short antisense compounds effectively inhibit the activity of DGAT2 or inhibit the expression of DGAT2. In one embodiment, the activity or expression of DGAT2 in a subject is inhibited by at least 10%, by at least 20%, by at least 25%, by at least 30%, by at least 40%, by at least 50%, by at least 60%, by at least 70%, by at least 75%, by at least 80%, by at least 85%, by at least 90%, by at least 95%, by at least 98%, by at least 99%, or by 100%. In certain embodiments, the activity or expression of DGAT2 in a subject is inhibited by about 30%. More preferably, the activity or expression of DGAT2 in a subject is inhibited by 50% or more.

The reduction of the expression of DGAT2 may be measured, for example, in blood, plasma, serum, adipose tissue, liver or any other body fluid, tissue or organ of the animal. Preferably, the cells contained within said fluids, tissues or organs being analyzed contain a nucleic acid molecule encoding DGAT2 and/or the DGAT2 protein itself.

In certain embodiments, pharmaceutical and other compositions comprising the compounds of the invention are also provided. For example, short antisense compounds targeted to a DGAT2 nucleic acid can be utilized in pharmaceutical compositions by adding an effective amount of a compound to a suitable pharmaceutically acceptable diluent or carrier.

Certain short antisense compounds targeting DGAT2 may have any one or more properties or characteristics of the short antisense compounds generally described herein. In certain embodiments, short antisense compounds targeting a DGAT2 nucleic acid have a motif (wing - deoxy gap -wing) selected 6om 1-12-1,1-1-10-2,2-10-1-1,3-10-3,2-10-3,2-10-2,1-10-1,1-10-2, 3-8-3,2-8-2,1-8-1,3-63 or 1-6-1. In certain embodiments, short antisense compounds targeting a DGAT2 nucleic acid have a motif (wing - deoxy gap -wing) selected from 1-10-1,2-10-2 and 3-10-3.

Provided herein are methods of treating an individual by administering one or more short antisense compound targeted to a DGAT2 nucleic acid or a pharmaceutical composition comprising such compound. Further provided are methods of treating a subject having a disease or conditions associated with DGAT2 activity by administering a short antisense compound targeted to a DGAT2 nucleic acid. Diseases and conditions associated with DGAT2 include, but are not limited to, cardiovascular disorders, obesity, diabetes, cholesterolemia, and liver steatosis.

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Certain Short Antisense Compounds Targeted to a DGAT2 Nucleic Acid

In certain embodiments, short antisense compounds are targeted to a DGAT2 nucleic acid having the sequence of GENBANK® Accession No. NM 032564.2, incoiporated herein as SEQ ID NO: 10. In certain such embodiments, a short antisense compound targeted to SEQ ID NO: 10 is at least 90% complementary to SEQ ID NO: 10. In certain such embodiments, a short antisense compound targeted to SEQ ID NO: 10 is at least 95% complementary to SEQ ID NO: 10. In certain such embodiments, a short antisense compound targeted to SEQ ID NO: 10 is 100% complementary to SEQ ID NO: 10. In certain embodiments, a short antisense compound targeted to SEQ ID NO: 10 includes a nucleotide sequence selected from the nucleotide sequences set forth in Tables 14 and 15.

Each nucleotide sequence set forth in each Tables 14 and 15 is independent of any modification to a sugar moiety, an intemucleoside linkage, or a nucleobase. As such, short antisense compounds comprising a nucleotide sequence as set forth in Tables 14 and 15 may comprise, independently, one or more modifications to a sugar moiety, an intemucleoside linkage, or a nucleobase. Antisense compounds described by Isis Number (Isis NO.) indicate a combination of nucleobase sequence and one or more modifications to a sugar moiety, an intemucleoside linkage, or a nucleobase.

Tables 14 and 15 illustrate examples of short antisense compounds targeted to SEQ ID NO: 10. Table 14 illustrates short antisense compounds that are 100% complementary to SEQ ID NO: 10. Table 15 illustrates short antisense compounds that have one or two mismatches with respect to SEQ ID NO: 10. The column labeled ‘gapmer motif indicates the wing-gap-wing motif of each short antisense compounds. The gap segment comprises 2’-deoxynucleotides and each nucleotide of each wing segment comprises a 2’-modified sugar. The particular 2’-modified sugar is also indicated in the ‘gapmer motif column. For example, ‘2-10-2 MOE’ means a 2-10-2 gapmer motif, where a gap segment of ten 2’deoxynucleotides is flanked by wing segments of two nucleotides, where the nucleotides of the wing segments are 2’-MOE nucleotides. Intemucleoside linkages are phosphorothioate. The short antisense compounds comprise 5-methylcytidine in place of unmodified cytosine, unless “unmodified cytosine” is listed in the gapmer motif column, in which case the indicated cytosines are unmodified cytosines. For example, “5-mC in gap only” indicates that the gap segment has 5-methylcytosines, while the wing segments have unmodified cytosines.

Table 14: Short Antisense Compounds targeted to SEQ ID NO: 10

<td> ISIS NO.</td><td> 5’ Target Site</td><td> 3’ Target Site</td><td> Sequence (5*-3*)</td><td> Gapmer Motif</td><td> SEQ ID NO</td>

<td> 372556</td><td> 231</td><td> 244</td><td> ATGAGGGTCTTCAT</td><td> 2-10-2 MOE</td><td> 681</td>

<td> 372557</td><td> 249</td><td> 262</td><td> ACCCCGGAGTAGGC</td><td> 2-10-2 MOE</td><td> 682</td>

<td> 382601</td><td> 249</td><td> 260</td><td> CCCGGAGTAGGC</td><td> 1-10-1 MOE</td><td> 683</td>

<td> 372480</td><td> 251</td><td> 266</td><td> CAGGACCCCGGAGTAG</td><td> 3-10-3 MOE</td><td> 684</td>

<td> 372481</td><td> 252</td><td> 267</td><td> GCAGGACCCCGGAGTA</td><td> 3-10-3 MOE</td><td> 685</td>

<td> 372558</td><td> 252</td><td> 265</td><td> AGGACCCCGGAGTA</td><td> 2-10-2 MOE</td><td> 686</td>

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<td> 372559</td><td> 253</td><td> 266</td><td> CAGGACCCCGGAGT</td><td> 2-10-2 MOE</td><td> 687</td>

<td> 382603</td><td> 331</td><td> 342</td><td> CAGACCCCTCGC</td><td> 1-10-1 MOE</td><td> 688</td>

<td> 382604</td><td> 361</td><td> 372</td><td> AGAGGATGCTGG</td><td> 1-10-1 MOE</td><td> 689</td>

<td> 372485</td><td> 392</td><td> 407</td><td> GAGCCAGGTGACAGAG</td><td> 3-10-3 MOE</td><td> 690</td>

<td> 372563</td><td> 393</td><td> 406</td><td> AGCCAGGTGACAGA</td><td> 2-10-2 MOE</td><td> 691</td>

<td> 382605</td><td> 397</td><td> 408</td><td> TGAGCCAGGTGA</td><td> 1-10-1 MOE</td><td> 692</td>

<td> 372565</td><td> 414</td><td> 427</td><td> TTTTCCACCTTGGA</td><td> 2-10-2 MOE</td><td> 693</td>

<td> 382606</td><td> 482</td><td> 493</td><td> CTGCAGGCCACT</td><td> 1-10-1 MOE</td><td> 694</td>

<td> 372497</td><td> 651</td><td> 666</td><td> TCACCAGCTGGATGGG</td><td> 3-10-3 MOE</td><td> 695</td>

<td> 372498</td><td> 652</td><td> 667</td><td> TTCACCAGCTGGATGG</td><td> 3-10-3 MOE</td><td> 696</td>

<td> 372575</td><td> 652</td><td> 665</td><td> CACCAGCTGGATGG</td><td> 2-10-2 MOE</td><td> 697</td>

<td> 372576</td><td> 653</td><td> 666</td><td> TCACCAGCTGGATG</td><td> 2-10-2 MOE</td><td> 698</td>

<td> 382607</td><td> 655</td><td> 666</td><td> TCACCAGCTGGA</td><td> 1-10-1 MOE</td><td> 699</td>

<td> 372499</td><td> 656</td><td> 671</td><td> TGTCTTCACCAGCTGG</td><td> 3-10-3 MOE</td><td> 700</td>

<td> 372577</td><td> 657</td><td> 670</td><td> GTCTTCACCAGCTG</td><td> 2-10-2 MOE</td><td> 701</td>

<td> 372500</td><td> 659</td><td> 674</td><td> GTGTGTCTTCACCAGC</td><td> 3-10-3 MOE</td><td> 702</td>

<td> 372578</td><td> 660</td><td> 673</td><td> TGTGTCTTCACCAG</td><td> 2-10-2 MOE</td><td> 703</td>

<td> 372501</td><td> 661</td><td> 676</td><td> TTGTGTGTCTTCACCA</td><td> 3-10-3 MOE</td><td> 704</td>

<td> 372579</td><td> 662</td><td> 675</td><td> TGTGTGTCTTCACC</td><td> 2-10-2 MOE</td><td> 705</td>

<td> 372502</td><td> 664</td><td> 679</td><td> AGGTTGTGTGTCTTCA</td><td> 3-10-3 MOE</td><td> 706</td>

<td> 372580</td><td> 665</td><td> 678</td><td> GGTTGTGTGTCTTC</td><td> 2-10-2 MOE</td><td> 707</td>

<td> 372503</td><td> 666</td><td> 681</td><td> GCAGGTTGTGTGTCTT</td><td> 3-10-3 MOE</td><td> 708</td>

<td> 372581</td><td> 667</td><td> 680</td><td> CAGGTTGTGTGTCT</td><td> 2-10-2 MOE</td><td> 709</td>

<td> 372504</td><td> 669</td><td> 684</td><td> TCAGCAGGTTGTGTGT</td><td> 3-10-3 MOE</td><td> 710</td>

<td> 372582</td><td> 670</td><td> 683</td><td> CAGCAGGTTGTGTG</td><td> 2-10-2 MOE</td><td> 711</td>

<td> 372505</td><td> 671</td><td> 686</td><td> GGTCAGCAGGTTGTGT</td><td> 3-10-3 MOE</td><td> 712</td>

<td> 372506</td><td> 672</td><td> 687</td><td> TGGTCAGCAGGTTGTG</td><td> 3-10-3 MOE</td><td> 713</td>

<td> 372583</td><td> 672</td><td> 685</td><td> GTCAGCAGGTTGTG</td><td> 2-10-2 MOE</td><td> 714</td>

<td> 372584</td><td> 673</td><td> 686</td><td> GGTCAGCAGGTTGT</td><td> 2-10-2 MOE</td><td> 715</td>

<td> 372507</td><td> 676</td><td> 691</td><td> CTGGTGGTCAGCAGGT</td><td> 3-10-3 MOE</td><td> 716</td>

<td> 372585</td><td> 677</td><td> 690</td><td> TGGTGGTCAGCAGG</td><td> 2-10-2 MOE</td><td> 717</td>

<td> 382608</td><td> 680</td><td> 691</td><td> CTGGTGGTCAGC</td><td> 1-10-1 MOE</td><td> 718</td>

<td> 372508</td><td> 681</td><td> 696</td><td> AGTTCCTGGTGGTCAG</td><td> 3-10-3 MOE</td><td> 719</td>

<td> 372586</td><td> 682</td><td> 695</td><td> GTTCCTGGTGGTCA</td><td> 2-10-2 MOE</td><td> 720</td>

<td> 372509</td><td> 684</td><td> 699</td><td> TATAGTTCCTGGTGGT</td><td> 3-10-3 MOE</td><td> 721</td>

<td> 372587</td><td> 685</td><td> 698</td><td> ATAGTTCCTGGTGG</td><td> 2-10-2 MOE</td><td> 722</td>

<td> 372510</td><td> 686</td><td> 701</td><td> GATATAGTTCCTGGTG</td><td> 3-10-3 MOE</td><td> 723</td>

<td> 372588</td><td> 687</td><td> 700</td><td> ATATAGTTCCTGGT</td><td> 2-10-2 MOE</td><td> 724</td>

<td> 372511</td><td> 691</td><td> 706</td><td> CCAAAGATATAGTTCC</td><td> 3-10-3 MOE</td><td> 725</td>

<td> 372512</td><td> 692</td><td> 707</td><td> TCCAAAGATATAGTTC</td><td> 3-10-3 MOE</td><td> 726</td>

<td> 372589</td><td> 692</td><td> 705</td><td> CAAAGATATAGTTC</td><td> 2-10-2 MOE</td><td> 727</td>

<td> 372590</td><td> 693</td><td> 706</td><td> CCAAAGATATAGTT</td><td> 2-10-2 MOE</td><td> 728</td>

<td> 382609</td><td> 724</td><td> 735</td><td> CCAGGCCCATGA</td><td> 1-10-1 MOE</td><td> 729</td>

<td> 372514</td><td> 725</td><td> 740</td><td> GGCACCCAGGCCCATG</td><td> 3-10-3 MOE</td><td> 730</td>

<td> 372592</td><td> 726</td><td> 739</td><td> GCACCCAGGCCCAT</td><td> 2-10-2 MOE</td><td> 731</td>

<td> 372515</td><td> 730</td><td> 745</td><td> CAGAAGGCACCCAGGC</td><td> 3-10-3 MOE</td><td> 732</td>

<td> 372593</td><td> 731</td><td> 744</td><td> AGAAGGCACCCAGG</td><td> 2-10-2 MOE</td><td> 733</td>

<td> 382610</td><td> 851</td><td> 862</td><td> CCAGACATCAGG</td><td> 1-10-1 MOE</td><td> 734</td>

<td> 382611</td><td> 867</td><td> 878</td><td> GACAGGGCAGAT</td><td> 1-10-1 MOE</td><td> 735</td>

<td> 382602</td><td> 868</td><td> 879</td><td> TGACAGGGCAGA</td><td> 1-10-1 MOE</td><td> 736</td>

<td> 382612</td><td> 911</td><td> 922</td><td> CCACTCCCATTC</td><td> 1-10-1 MOE</td><td> 737</td>

<td> 372524</td><td> 965</td><td> 980</td><td> GCCAGGCATGGAGCTC</td><td> 3-10-3 MOE</td><td> 738</td>

<td> 372602</td><td> 966</td><td> 979</td><td> CCAGGCATGGAGCT</td><td> 2-10-2 MOE</td><td> 739</td>

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<td> 382613</td><td> 968</td><td> 979</td><td> CCAGGCATGGAG</td><td> 1-10-1 MOE</td><td> 740</td>

<td> 382614</td><td> 987</td><td> 998</td><td> CAGGGTGACTGC</td><td> 1-10-1 MOE</td><td> 741</td>

<td> 372525</td><td> 989</td><td> 1004</td><td> GTTCCGCAGGGTGACT</td><td> 3-10-3 MOE</td><td> 742</td>

<td> 372603</td><td> 990</td><td> 1003</td><td> TTCCGCAGGGTGAC</td><td> 2-10-2 MOE</td><td> 743</td>

<td> 372526</td><td> 992</td><td> 1007</td><td> GCGGTTCCGCAGGGTG</td><td> 3-10-3 MOE</td><td> 744</td>

<td> 372604</td><td> 993</td><td> 1006</td><td> CGGTTCCGCAGGGT</td><td> 2-10-2 MOE</td><td> 745</td>

<td> 372530</td><td> 1106</td><td> 1121</td><td> TCGGCCCCAGGAGCCC</td><td> 3-10-3 MOE</td><td> 746</td>

<td> 372608</td><td> 1107</td><td> 1120</td><td> CGGCCCCAGGAGCC</td><td> 2-10-2 MOE</td><td> 747</td>

<td> 372531</td><td> 1109</td><td> 1124</td><td> CCATCGGCCCCAGGAG</td><td> 3-10-3 MOE</td><td> 748</td>

<td> 372609</td><td> 1110</td><td> 1123</td><td> CATCGGCCCCAGGA</td><td> 2-10-2 MOE</td><td> 749</td>

<td> 372532</td><td> 1112</td><td> 1127</td><td> GACCCATCGGCCCCAG</td><td> 3-10-3 MOE</td><td> 750</td>

<td> 372610</td><td> 1113</td><td> 1126</td><td> ACCCATCGGCCCCA</td><td> 2-10-2 MOE</td><td> 751</td>

<td> 372533</td><td> 1117</td><td> 1132</td><td> TTCTGGACCCATCGGC</td><td> 3-10-3 MOE</td><td> 752</td>

<td> 382615</td><td> 1117</td><td> 1128</td><td> GGACCCATCGGC</td><td> 1-10-1 MOE</td><td> 753</td>

<td> 372611</td><td> 1118</td><td> 1131</td><td> TCTGGACCCATCGG</td><td> 2-10-2 MOE</td><td> 754</td>

<td> 372536</td><td> 1199</td><td> 1214</td><td> CACCAGCCCCCAGGTG</td><td> 3-10-3 MOE</td><td> 755</td>

<td> 372614</td><td> 1200</td><td> 1213</td><td> ACCAGCCCCCAGGT</td><td> 2-10-2 MOE</td><td> 756</td>

<td> 372537</td><td> 1204</td><td> 1219</td><td> TAGGGCACCAGCCCCC</td><td> 3-10-3 MOE</td><td> 757</td>

<td> 372615</td><td> 1205</td><td> 1218</td><td> AGGGCACCAGCCCC</td><td> 2-10-2 MOE</td><td> 758</td>

<td> 372538</td><td> 1209</td><td> 1224</td><td> TGGAGTAGGGCACCAG</td><td> 3-10-3 MOE</td><td> 759</td>

<td> 372616</td><td> 1210</td><td> 1223</td><td> GGAGTAGGGCACCA</td><td> 2-10-2 MOE</td><td> 760</td>

<td> 382616</td><td> 1215</td><td> 1226</td><td> CTTGGAGTAGGG</td><td> 1-10-1 MOE</td><td> 761</td>

<td> 372539</td><td> 1218</td><td> 1233</td><td> TGATGGGCTTGGAGTA</td><td> 3-10-3 MOE</td><td> 762</td>

<td> 372617</td><td> 1219</td><td> 1232</td><td> GATGGGCTTGGAGT</td><td> 2-10-2 MOE</td><td> 763</td>

<td> 372540</td><td> 1293</td><td> 1308</td><td> TGTGGTACAGGTCGAT</td><td> 3-10-3 MOE</td><td> 764</td>

<td> 372618</td><td> 1294</td><td> 1307</td><td> GTGGTACAGGTCGA</td><td> 2-10-2 MOE</td><td> 765</td>

<td> 382617</td><td> 1294</td><td> 1305</td><td> GGTACAGGTCGA</td><td> 1-10-1 MOE</td><td> 766</td>

<td> 372541</td><td> 1295</td><td> 1310</td><td> GGTGTGGTACAGGTCG</td><td> 3-10-3 MOE</td><td> 767</td>

<td> 372619</td><td> 1296</td><td> 1309</td><td> GTGTGGTACAGGTC</td><td> 2-10-2 MOE</td><td> 768</td>

<td> 372542</td><td> 1298</td><td> 1313</td><td> CATGGTGTGGTACAGG</td><td> 3-10-3 MOE</td><td> 769</td>

<td> 372620</td><td> 1299</td><td> 1312</td><td> ATGGTGTGGTACAG</td><td> 2-10-2 MOE</td><td> 770</td>

<td> 372543</td><td> 1300</td><td> 1315</td><td> TACATGGTGTGGTACA</td><td> 3-10-3 MOE</td><td> 771</td>

<td> 372621</td><td> 1301</td><td> 1314</td><td> ACATGGTGTGGTAC</td><td> 2-10-2 MOE</td><td> 772</td>

<td> 372544</td><td> 1303</td><td> 1318</td><td> ATGTACATGGTGTGGT</td><td> 3-10-3 MOE</td><td> 773</td>

<td> 372622</td><td> 1304</td><td> 1317</td><td> TGTACATGGTGTGG</td><td> 2-10-2 MOE</td><td> 774</td>

<td> 382618</td><td> 1313</td><td> 1324</td><td> GCCTCCATGTAC</td><td> 1-10-1 MOE</td><td> 775</td>

<td> 382619</td><td> 1325</td><td> 1336</td><td> AGCTTCACCAGG</td><td> 1-10-1 MOE</td><td> 776</td>

<td> 382620</td><td> 1383</td><td> 1394</td><td> GTTCACCTCCAG</td><td> 1-10-1 MOE</td><td> 777</td>

Table 15: Short antisense compounds targeted to SEQ ID NO: 10 and having 1 or 2 mismatches

<td> ISIS NO</td><td> 5’ Target Site</td><td> 3’ Target Site</td><td> Sequence (5’-3’)</td><td> Gapmer Motif</td><td> SEQ IDNO</td>

<td> 372608</td><td> 151</td><td> 164</td><td> CGGCCCCAGGAGCC</td><td> 2-10-2 MOE</td><td> 747</td>

<td> 372474</td><td> 156</td><td> 171</td><td> CATGCCCCAGCCGCCG</td><td> 3-10-3 MOE</td><td> 778</td>

<td> 372552</td><td> 157</td><td> 170</td><td> ATGCCCCAGCCGCC</td><td> 2-10-2 MOE</td><td> 779</td>

<td> 382609</td><td> 167</td><td> 178</td><td> CCAGGCCCATGA</td><td> 1-10-1 MOE</td><td> 729</td>

<td> 372478</td><td> 230</td><td> 245</td><td> GATGAGGGTCTTCATG</td><td> 3-10-3 MOE</td><td> 780</td>

<td> 372479</td><td> 248</td><td> 263</td><td> GACCCCGGAGTAGGCA</td><td> 3-10-3 MOE</td><td> 781</td>

<td> 382611</td><td> 317</td><td> 328</td><td> GACAGGGCAGAT</td><td> 1-10-1 MOE</td><td> 735</td>

<td> 372483</td><td> 352</td><td> 367</td><td> ATGCTGGAGCCAGTGC</td><td> 3-10-3 MOE</td><td> 782</td>

<td> 372561</td><td> 353</td><td> 366</td><td> TGCTGGAGCCAGTG</td><td> 2-10-2 MOE</td><td> 783</td>

<td> 372562</td><td> 373</td><td> 386</td><td> GTCTTGGAGGGCCG</td><td> 2-10-2 MOE</td><td> 784</td>

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<td> 382602</td><td> 388</td><td> 399</td><td> TGACAGGGCAGA</td><td> 1-10-1 MOE</td><td> 736</td>

<td> 372613</td><td> 392</td><td> 405</td><td> CCCAGGTGTCAGAG</td><td> 2-10-2 MOE</td><td> 785</td>

<td> 372486</td><td> 412</td><td> 427</td><td> TTTTCCACCTTGGATC</td><td> 3-10-3 MOE</td><td> 786</td>

<td> 372564</td><td> 413</td><td> 426</td><td> TTTCCACCTTGGAT</td><td> 2-10-2 MOE</td><td> 787</td>

<td> 372487</td><td> 413</td><td> 428</td><td> TTTTTCCACCTTGGAT</td><td> 3-10-3 MOE</td><td> 788</td>

<td> 372488</td><td> 418</td><td> 433</td><td> AGGTGTTTTTCCACCT</td><td> 3-10-3 MOE</td><td> 789</td>

<td> 372566</td><td> 419</td><td> 432</td><td> GGTGTTTTTCCACC</td><td> 2-10-2 MOE</td><td> 790</td>

<td> 372489</td><td> 459</td><td> 474</td><td> CCAGGAAGGATAGGAC</td><td> 3-10-3 MOE</td><td> 791</td>

<td> 372567</td><td> 460</td><td> 473</td><td> CAGGAAGGATAGGA</td><td> 2-10-2 MOE</td><td> 792</td>

<td> 382612</td><td> 475</td><td> 486</td><td> CCACTCCCATTC</td><td> 1-10-1 MOE</td><td> 737</td>

<td> 372490</td><td> 483</td><td> 498</td><td> TGACACTGCAGGCCAC</td><td> 3-10-3 MOE</td><td> 793</td>

<td> 372568</td><td> 484</td><td> 497</td><td> GACACTGCAGGCCA</td><td> 2-10-2 MOE</td><td> 794</td>

<td> 372491</td><td> 492</td><td> 507</td><td> ACATGAGGATGACACT</td><td> 3-10-3 MOE</td><td> 795</td>

<td> 372569</td><td> 493</td><td> 506</td><td> CATGAGGATGACAC</td><td> 2-10-2 MOE</td><td> 796</td>

<td> 372492</td><td> 503</td><td> 518</td><td> GCAGAAGGTGTACATG</td><td> 3-10-3 MOE</td><td> 797</td>

<td> 372570</td><td> 504</td><td> 517</td><td> CAGAAGGTGTACAT</td><td> 2-10-2 MOE</td><td> 798</td>

<td> 372493</td><td> 512</td><td> 527</td><td> GCAGTCAGTGCAGAAG</td><td> 3-10-3 MOE</td><td> 799</td>

<td> 372571</td><td> 513</td><td> 526</td><td> CAGTCAGTGCAGAA</td><td> 2-10-2 MOE</td><td> 800</td>

<td> 372496</td><td> 612</td><td> 627</td><td> ACACGGCCCAGTTTCG</td><td> 3-10-3 MOE</td><td> 801</td>

<td> 372574</td><td> 613</td><td> 626</td><td> CACGGCCCAGTTTC</td><td> 2-10-2 MOE</td><td> 802</td>

<td> 372513</td><td> 717</td><td> 732</td><td> GGCCCATGATGCCATG</td><td> 3-10-3 MOE</td><td> 803</td>

<td> 372591</td><td> 718</td><td> 731</td><td> GCCCATGATGCCAT</td><td> 2-10-2 MOE</td><td> 804</td>

<td> 372516</td><td> 732</td><td> 747</td><td> TACAGAAGGCACCCAG</td><td> 3-10-3 MOE</td><td> 805</td>

<td> 372594</td><td> 733</td><td> 746</td><td> ACAGAAGGCACCCA</td><td> 2-10-2 MOE</td><td> 806</td>

<td> 372518</td><td> 812</td><td> 827</td><td> GAAGTTGCCAGCCAAT</td><td> 3-10-3 MOE</td><td> 807</td>

<td> 372596</td><td> 813</td><td> 826</td><td> AAGTTGCCAGCCAA</td><td> 2-10-2 MOE</td><td> 808</td>

<td> 372560</td><td> 863</td><td> 876</td><td> CAGGGCAGATCCTT</td><td> 2-10-2 MOE</td><td> 809</td>

<td> 372519</td><td> 887</td><td> 902</td><td> CAAGTAGTCTATGGTG</td><td> 3-10-3 MOE</td><td> 810</td>

<td> 372597</td><td> 888</td><td> 901</td><td> AAGTAGTCTATGGT</td><td> 2-10-2 MOE</td><td> 811</td>

<td> 372520</td><td> 894</td><td> 909</td><td> TGGAAAGCAAGTAGTC</td><td> 3-10-3 MOE</td><td> 812</td>

<td> 372598</td><td> 895</td><td> 908</td><td> GGAAAGCAAGTAGT</td><td> 2-10-2 MOE</td><td> 813</td>

<td> 372527</td><td> 1013</td><td> 1028</td><td> GGCCAGCTTTACAAAG</td><td> 3-10-3 MOE</td><td> 814</td>

<td> 372605</td><td> 1014</td><td> 1027</td><td> GCCAGCTTTACAAA</td><td> 2-10-2 MOE</td><td> 815</td>

<td> 372606</td><td> 1020</td><td> 1033</td><td> CGCAGGGCCAGCTT</td><td> 2-10-2 MOE</td><td> 816</td>

<td> 372529</td><td> 1052</td><td> 1067</td><td> AAAGGAATAGGTGGGA</td><td> 3-10-3 MOE</td><td> 817</td>

<td> 372607</td><td> 1053</td><td> 1066</td><td> AAGGAATAGGTGGG</td><td> 2-10-2 MOE</td><td> 818</td>

<td> 372534</td><td> 1144</td><td> 1159</td><td> GCGAAACCAATATACT</td><td> 3-10-3 MOE</td><td> 819</td>

<td> 372612</td><td> 1145</td><td> 1158</td><td> CGAAACCAATATAC</td><td> 2-10-2 MOE</td><td> 820</td>

<td> 372535</td><td> 1192</td><td> 1207</td><td> CCCCAGGTGTCAGAGG</td><td> 3-10-3 MOE</td><td> 821</td>

<td> 372613</td><td> 1193</td><td> 1206</td><td> CCCAGGTGTCAGAG</td><td> 2-10-2 MOE</td><td> 822</td>

<td> 372545</td><td> 1332</td><td> 1347</td><td> GATTGTCAAAGAGCTT</td><td> 3-10-3 MOE</td><td> 823</td>

<td> 372623</td><td> 1333</td><td> 1346</td><td> ATTGTCAAAGAGCT</td><td> 2-10-2 MOE</td><td> 824</td>

<td> 372546</td><td> 1342</td><td> 1357</td><td> TTGGTCTTGTGATTGT</td><td> 3-10-3 MOE</td><td> 825</td>

<td> 372624</td><td> 1343</td><td> 1356</td><td> TGGTCTTGTGATTG</td><td> 2-10-2 MOE</td><td> 826</td>

<td> 372547</td><td> 1352</td><td> 1367</td><td> AAGGCCGAATTTGGTC</td><td> 3-10-3 MOE</td><td> 827</td>

<td> 372625</td><td> 1353</td><td> 1366</td><td> AGGCCGAATTTGGT</td><td> 2-10-2 MOE</td><td> 828</td>

<td> 382601</td><td> 1617</td><td> 1628</td><td> CCCGGAGTAGGC</td><td> 1-10-1 MOE</td><td> 683</td>

<td> 382606</td><td> 1971</td><td> 1982</td><td> CTGCAGGCCACT</td><td> 1-10-1 MOE</td><td> 694</td>

<td> 382612</td><td> 1988</td><td> 1999</td><td> CCACTCCCATTC</td><td> 1-10-1 MOE</td><td> 737</td>

In certain embodiments, a target region is nucleotides 231-267 of SEQ ID NO: 10. In certain embodiments, a short antisense compound is targeted to nucleotides 231-267 of SEQ ID NO: 10. In

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-135PCI7US2007/068401 certain such embodiments, a short antisense compound targeted to nucleotides 231-267 comprises a nucleotide sequence selected from SEQ ID NO 681, 682, 683, 684, 685, 686, or 687. In certain such embodiments, a short antisense compound targeted to nucleotides 231-267 of SEQ ID NO: 10 is selected from Isis No 372556,372557,382601,372480,372481,372558, or 372559.

In certain embodiments, a target region is nucleotides 249-267 of SEQ ID NO: 10. In certain embodiments, a short antisense compound is targeted to nucleotides 249-267 of SEQ ID NO: 10. In certain such embodiments, a short antisense compound targeted to nucleotides 249-267 comprises a nucleotide sequence selected from SEQ ID NO 683,684,685,686, or 687. In certain such embodiments, a short antisense compound targeted to nucleotides 249-267 of SEQ ID NO: 10 is selected from Isis No 382601,372480,372481,372558, or 372559.

In certain embodiments, a target region is nucleotides 331-493 of SEQ ID NO: 10. In certain embodiments, a short antisense compound is targeted to nucleotides 331-493 of SEQ ID NO: 10. In certain such embodiments, a short antisense compound targeted to nucleotides 331-493 comprises a nucleotide sequence selected from SEQ ID NO 688, 689, 690, 691, 692, 693, or 694. In certain such embodiments, a short antisense compound targeted to nucleotides 331-493 of SEQ ID NO: 10 is selected from Isis No 382603,382604,372485,372563,382605,372565, or 382606.

In certain embodiments, a target region is nucleotides 331-427 of SEQ ID NO: 10. In certain embodiments, a short antisense compound is targeted to nucleotides 331-427 of SEQ ID NO: 10. In certain such embodiments, a short antisense compound targeted to nucleotides 331-427 comprises a nucleotide sequence selected from SEQ ID NO 688, 689, 690, 691, 692, or 693. In certain such embodiments, a short antisense compound targeted to nucleotides 331-427 of SEQ ID NO: 10 is selected from Isis No 382603,382604,372485,372563,382605, or 372565.

In certain embodiments, a target region is nucleotides 392-408 of SEQ ID NO: 10. In certain embodiments, a short antisense compound is targeted to nucleotides 392-408 of SEQ ID NO: 10. In certain such embodiments, a short antisense compound targeted to nucleotides 392-408 comprises a nucleotide sequence selected from SEQ ID NO 690, 691, or 692. In certain such embodiments, a short antisense compound targeted to nucleotides 392-408 of SEQ ID NO: 10 is selected from Isis No 372485, 372563, or 382605.

In certain embodiments, a target region is nucleotides 651-707 of SEQ ID NO: 10. In certain embodiments, a short antisense compound is targeted to nucleotides 651-707 of SEQ ID NO: 10. In certain such embodiments, a short antisense compound targeted to nucleotides 651-707 comprises a nucleotide sequence selected from SEQ ID NO 695, 696, 697, 698, 699, 700, 701, 702, 703, 704, 705, 706,707, 708,709, 710,711,712,713,714, 715,716,717,718,719,720,721,722,723, 724,725,726, 727, or 728. In certain such embodiments, a short antisense compound targeted to nucleotides 651-707 of SEQ ID NO: 10 is selected from Isis No 372497, 372498, 372575, 372576, 382607, 372499, 372577, 372500, 372578, 372501, 372579,372502,372580,372503, 372581, 372504, 372582, 372505, 372506, 372583, 372584, 372507, 372585, 382608, 372508, 372586, 372509, 372587, 372510, 372588, 372511,

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-136372512,372589, or 372590.

In certain embodiments, a target region is nucleotides 724-745 of SEQ ID NO: 10. In certain embodiments, a short antisense compound is targeted to nucleotides 724-745 of SEQ ID NO: 10. In certain such embodiments, a short antisense compound targeted to nucleotides 724-745 comprises a nucleotide sequence selected from SEQ ID NO 729,730,731,732, or 733. In certain such embodiments, a short antisense compound targeted to nucleotides 724-745 of SEQ ID NO: 10 is selected from Isis No

382609.372514.372592.372515, or 372593.

In certain embodiments, a target region is nucleotides 651-745 of SEQ ID NO: 10. In certain embodiments, a short antisense compound is targeted to nucleotides 651-745 of SEQ ID NO; 10. In certain such embodiments, a short antisense compound targeted to nucleotides 651-745 comprises a nucleotide sequence selected from SEQ ID NO 695, 696, 697, 698, 699, 700, 701, 702, 703, 704, 705, 706,707,708,709,710,711,712,713,714,715,716,717,718,719,720,721,722,723,724,725,726, 727, 728,729,730, 731,732, or 733. In certain such embodiments, a short antisense compound targeted to nucleotides 651-745 of SEQ ID NO: 10 is selected from Isis No 372497, 372498, 372575, 372576, 382607, 372499, 372577, 372500, 372578,372501,372579,372502, 372580, 372503, 372581, 372504, 372582, 372505, 372506, 372583, 372584, 372507, 372585,382608, 372508, 372586, 372509, 372587,

372510.372588.372511.372512.372589.372590.382609.372514.372592.372515, or 372593.

In certain embodiments, a target region is nucleotides 851-922 of SEQ ID NO: 10. In certain embodiments, a short antisense compound is targeted to nucleotides 851-922 of SEQ ID NO: 10. In certain such embodiments, a short antisense compound targeted to nucleotides 851-922 comprises a nucleotide sequence selected from SEQ ID NO 734, 735, 736, or 737. In certain such embodiments, a short antisense compound targeted to nucleotides 851-922 of SEQ ID NO: 10 is selected from Isis No 382610,382611,382602, or 382612.

In certain embodiments, a target region is nucleotides 851-879 of SEQ ID NO: 10. In certain embodiments, a short antisense compound is targeted to nucleotides 851-879 of SEQ ID NO: 10. In certain such embodiments, a short antisense compound targeted to nucleotides 851-879 comprises a nucleotide sequence selected from SEQ ID NO 734, 735, or 736. In certain such embodiments, a short antisense compound targeted to nucleotides 851-879 of SEQ ID NO: 10 is selected from Isis No 382610, 382611, or 382602.

In certain embodiments, a target region is nucleotides 965-1007 of SEQ ID NO: 10. In certain embodiments, a short antisense compound is targeted to nucleotides 965-1007 of SEQ ID NO: 10. In certain such embodiments, a short antisense compound targeted to nucleotides 965-1007 comprises a nucleotide sequence selected from SEQ ID NO 738,739,740,741,742,743,744, or 745. In certain such embodiments, a short antisense compound targeted to nucleotides 965-1007 of SEQ ID NO: 10 is selected from Isis No 372524,372602, 382613, 382614,372525,372603,372526, or 372604.

In certain embodiments, a target region is nucleotides 965-979 of SEQ ID NO: 10. In certain embodiments, a short antisense compound is targeted to nucleotides 965-979 of SEQ ID NO: 10. In

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-137PCT/US2007/068401 certain such embodiments, a short antisense compound targeted to nucleotides 965-979 comprises a nucleotide sequence selected from SEQ ID NO 738, 739, or 740. In certain such embodiments, a short antisense compound targeted to nucleotides 965-979 of SEQ ID NO: 10 is selected from Isis No 372524, 372602, or 382613.

In certain embodiments, a target region is nucleotides 987-1007 of SEQ ID NO: 10. In certain embodiments, a short antisense compound is targeted to nucleotides 987-1007 of SEQ ID NO: 10. In certain such embodiments, a short antisense compound targeted to nucleotides 987-1007 comprises a nucleotide sequence selected from SEQ ID NO 741,742,743,744, or 745. In certain such embodiments, a short antisense compound targeted to nucleotides 987-1007 of SEQ ID NO: 10 is selected from Isis No 382614,372525,372603,372526, or 372604.

In certain embodiments, a target region is nucleotides 1106-1132 of SEQ ID NO: 10. In certain embodiments, a short antisense compound is targeted to nucleotides 1106-1132 of SEQ ID NO: 10. In certain such embodiments, a short antisense compound targeted to nucleotides 1106-1132 comprises a nucleotide sequence selected from SEQ ID NO 746,747,748,749,750,751,752,753, or 754. In certain such embodiments, a short antisense compound targeted to nucleotides 1106-1132 of SEQ ID NO: 10 is selected from Isis No 372530,372608,372531,372609,372532,372610,372533,382615, or 372611.

In certain embodiments, a target region is nucleotides 1199-1233 of SEQ ID NO: 10. In certain embodiments, a short antisense compound is targeted to nucleotides 1199-1233 of SEQ ID NO: 10. In certain such embodiments, a short antisense compound targeted to nucleotides 1199-1233 comprises a nucleotide sequence selected from SEQ ID NO 755,756,757,758,759,760,761,762, or 763. In certain such embodiments, a short antisense compound targeted to nucleotides 1199-1233 of SEQ ID NO: 10 is selected from Isis No 372536,372614,372537,372615,372538,372616, 382616,372539, or 372617.

In certain embodiments, a target region is nucleotides 1293-1394 of SEQ ID NO: 10. In certain embodiments, a short antisense compound is targeted to nucleotides 1293-1394 of SEQ ID NO: 10. In certain such embodiments, a short antisense compound targeted to nucleotides 1293-1394 comprises a nucleotide sequence selected from SEQ ID NO 764, 765, 766, 767, 768, 769, 770, 771, 772, 773, 774, 775,776, or 777. In certain such embodiments, a short antisense compound targeted to nucleotides 12931394 of SEQ ID NO: 10 is selected from Isis No 372540, 372618, 382617, 372541, 372619, 372542, 372620,372543,372621,372544,372622,382618,382619, or 382620.

In certain embodiments, a target region is nucleotides 1293-1336 of SEQ ID NO: 10. In certain embodiments, a short antisense compound is targeted to nucleotides 1293-1336 of SEQ ID NO: 10. In certain such embodiments, a short antisense compound targeted to nucleotides 1293-1336 comprises a nucleotide sequence selected from SEQ ID NO 764, 765, 766, 767, 768, 769, 770, 771, 772, 773, 774, 775, or 776. In certain such embodiments, a short antisense compound targeted to nucleotides 1293-1336 of SEQ ID NO: 10 is selected from Isis No 372540, 372618, 382617, 372541, 372619, 372542,372620, 372543,372621,372544,372622,382618, or 382619.

In certain embodiments, a target region is nucleotides 1293-1324 of SEQ ID NO: 10. In certain

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-138embodiments, a short antisense compound is targeted to nucleotides 1293-1324 of SEQ ID NO: 10. In certain such embodiments, a short antisense compound targeted to nucleotides 1293-1324 comprises a nucleotide sequence selected from SEQ ID NO 764,765,766,767,768,769,770,771,772,773,774, or 775. In certain such embodiments, a short antisense compound targeted to nucleotides 1293-1324 of SEQ ID NO: 10 is selected from Isis No 372540, 372618, 382617,372541, 372619, 372542, 372620, 372543, 372621,372544,372622, or 382618.

In certain embodiments, short antisense compounds targeted to a DGAT2 nucleic acid are 8 to 16, preferably 9 to 15, more preferably 9 to 14, more preferably 10 to 14 nucleotides in length. In certain embodiments, short antisense compounds targeted to a DGAT2 nucleic acid are 9 to 14 nucleotides in length. In certain embodiments, short antisense compounds targeted to a DGAT2 nucleic acid are 10 to 14 nucleotides in length. In certain embodiments, such short antisense compounds are short antisense oligonucleotides.

In certain embodiments, short antisense compounds targeted to a DGAT2 nucleic acid are short gapmers. In certain such embodiments, short gapmers targeted to a DGAT2 nucleic acid comprise at least one high affinity modification in one or more wings of the compound. In certain embodiments, short antisense compounds targeted to a DGAT2 nucleic acid comprise 1 to 3 high-affinity modifications in each wing. In certain such embodiments, the nucleosides or nucleotides of the wing comprise a 2’ modification. In certain such embodiments, the monomers of the wing are BNA’s. In certain such embodiments, the monomers of the wing are selected from α-L-Methyleneoxy (4’-CH_rO-2’) BNA, β-DMethyleneoxy (4’-CH₂-O-2’) BNA , Ethyleneoxy (4’-(CH₂)rO-2’) BNA , Aminooxy (4’-CH₂-O-N(R)2’) BNA and Oxyamino (4’-CH₂-N(R)-O-2’) BNA. In certain embodiments, the monomers of a wing comprise a substituent at the 2’ position selected from allyl, amino, azido, thio, O-allyl, 0-C|-Cio alkyl, OCFj, O-fCHjh-O-CHj, 2'-O(CH₂)₂SCHj,O-(CH₂)₂-O-N(R_ni)(R_n), and O-CH₂-C(=O)-N(R_K)(RJ, where each R„ and R„ is, independently, H or substituted or unsubstituted C_rC|o alkyl. In certain embodiments, the monomers of a wing are 2’MOE nucleotides.

In certain embodiments, short antisense compounds targeted to a DGAT2 nucleic acid comprise a gap between the 5’ wing and the 3’ wing. In certain embodiments the gap comprises five, six, seven, eight, nine, ten, eleven, twelve, thirteen, or fourteen monomers. In certain embodiments, the monomers of the gap are unmodified deoxyribonucleotides. In certain embodiments, the monomers of the gap are unmodified ribonucleotides. In certain embodiments, gap modifications (if any) gap result in a short antisense compound that, when bound to its target nucleic acid, supports cleavage by an RNase, including, but not limited to, RNase H.

In certain embodiments, short antisense compounds targeted to a DGAT2 nucleic acid have uniform monomeric linkages. In certain such embodiments, those linkages are all phosphorothioate linkages. In certain embodiments, the linkages are all phosphodlester linkages. In certain embodiments, short antisense compounds targeted to a DGAT2 nucleic acid have mixed backbones.

In certain embodiments, short antisense compounds targeted to a DGAT2 nucleic acid are 8

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-139monomers in length. In certain embodiments, short antisense compounds targeted to a DGAT2 nucleic acid are 9 monomers in length. In certain embodiments, short antisense compounds targeted to a DGAT2 nucleic acid are 10 monomers in length. In certain embodiments, short antisense compounds targeted to a DGAT2 nucleic acid are 11 monomers in length. In certain embodiments, short antisense compounds targeted to a DGAT2 nucleic acid are monomers in length. In certain embodiments, short antisense compounds targeted to a DGAT2 nucleic acid are 13 monomers in length. In certain embodiments, short antisense compounds targeted to a DGAT2 nucleic acid are 14 monomers in length. In certain embodiments, short antisense compounds targeted to a DGAT2 nucleic acid are 15 monomers in length. In certain embodiments, short antisense compounds targeted to a DGAT2 nucleic acid are 16 monomers in length. In certain embodiments, short antisense compounds targeted to a DGAT2 nucleic acid comprise 9 to 15 monomers. In certain embodiments, short antisense compounds targeted to a DGAT2 nucleic acid comprise 10 to 15 monomers. In certain embodiments, short antisense compounds targeted to a DGAT2 nucleic acid comprise 12 to 14 monomers. In certain embodiments, short antisense compounds targeted to a DGAT2 nucleic acid comprise 12 to 14 nucleotides or nucleosides.

In certain embodiments, the invention provides methods of modulating expression of DGAT2. In certain embodiments, such methods comprise use of one or more short antisense compound targeted to a DGAT2 nucleic acid, wherein the short antisense compound targeted to a DGAT2 nucleic acid is from about 8 to about 16, preferably 9 to 15, more preferably 9 to 14, more preferably 10 to 14 monomers (i.e. from about 8 to about 16 linked monomers). One of ordinary skill in the art will appreciate that this comprehends methods of modulating expression of DGAT2 using one or more short antisense compounds targeted to a DGAT2 nucleic acid of 8,9,10,11,12,13,14,15 or 16 monomers.

In certain embodiments, methods of modulating DGAT2 comprise use of a short antisense compound targeted to a DGAT2 nucleic acid that is 8 monomers in length In certain embodiments, methods of modulating DGAT2 comprise use of a short antisense compound targeted to a DGAT2 nucleic acid that is 9 monomers in length. In certain embodiments, methods of modulating DGAT2 comprise use of a short antisense compound targeted to a DGAT2 nucleic acid that is 10 monomers in length. In certain embodiments, methods of modulating DGAT2 comprise use of a short antisense compound targeted to a DGAT2 nucleic acid that is 11 monomers in length. In certain embodiments, methods of modulating DGAT2 comprise use of a short antisense compound targeted to a DGAT2 nucleic acid that is 12 monomers in length. In certain embodiments, methods of modulating DGAT2 comprise use of a short antisense compound targeted to a DGAT2 nucleic acid that is 13 monomers in length. In certain embodiments, methods of modulating DGAT2 comprise use of a short antisense compound targeted to a DGAT2 nucleic acid that is 14 monomers in length. In certain embodiments, methods of modulating DGAT2 comprise use of a short antisense compound targeted to a DGAT2 nucleic acid that is 15 monomers in length. In certain embodiments, methods of modulating DGAT2 comprise use of a short antisense compound targeted to a DGAT2 nucleic acid that is 16 monomers in length.

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-140In certain embodiments, methods of modulating expression of DGAT2 comprise use of a short antisense compound targeted to a DGAT2 nucleic acid comprising 9 to 15 monomers. In certain embodiments, methods of modulating expression of DGAT2 comprise use of a short antisense compound targeted to a DGAT2 nucleic acid comprising 10 to 15 monomers. In certain embodiments, methods of modulating expression of DGAT2 comprise use of a short antisense compound targeted to a DGAT2 nucleic acid comprising 12 to 14 monomers. In certain embodiments, methods of modulating expression of DGAT2 comprise use of a short antisense compound targeted to a DGAT2 nucleic acid comprising 12 or 14 nucleotides or nucleosides.

9. PTP1B

PTP1B (also known as protein phosphatase IB and PTPN1) is an endoplasmic reticulum (ER)associated enzyme originally isolated as the major protein tyrosine phosphatase of the human placenta (Tonks et al., J. Biol. Chem., 1988,263,6731 -6737; Tonks et al., J. Biol. Chem., 1988,263,6722-6730).

An essential regulatory role in signaling mediated by the insulin receptor has been established for PTP1B. In certain instances, PTP1B interacts with and dephosphorylates the activated insulin receptor both in vitro and in intact cells resulting in the downregulation of the signaling pathway (Goldstein et al., Mol. Cell. Biochem., 1998,182,91-99; Seely et al., Diabetes, 1996,45, 1379-1385). In addition, PTP1B modulates the mitogenic actions of insulin (Goldstein et al., Mol. Cell. Biochem., 1998,182,91-99). In rat adipose cells overexpressing PTP1B, the translocation of the GLUT4 glucose transporter was inhibited, implicating PTP1B as a negative regulator of glucose transport as well (Chen et al., J. Biol. Chem., 1997, 272,8026-8031).

Mouse knockout models lacking the PTP1B gene also point toward the negative regulation of insulin signaling by PTP1B. Mice harboring a disrupted PTP1B gene showed increased insulin sensitivity and increased phosphorylation of the insulin receptor. When placed on a high-fat diet, PTP1B -/- mice were resistant to weight gain and remained insulin sensitive (Elchebly el al., Science, 1999,283, 1544-1548). These studies clearly establish PTP1B as a therapeutic target in the treatment of diabetes and obesity.

Diabetes and obesity (sometimes now collectively referred to as “diabesity”) are interrelated. Most human obesity is associated with insulin resistance and leptin resistance. In fact obesity may have an even greater impact on insulin action than does diabetes itself (Sindelka et al., Physiol Res., 2002,57, 85-91). Syndrome X or metabolic syndrome is a new term for a cluster of conditions, that, when occurring together, may indicate a predisposition to diabetes and cardiovascular disease. These symptoms, including high blood pressure, high triglycerides, decreased HDL and obesity, tend to appear together in some individuals. Because of its role in both diabetes and obesity, PTP1B is believed to be a therapeutic target for a range of metabolic conditions, including diabetes, obesity and metabolic syndrome. By improving blood glucose control, inhibitors of PTP1B may also be useful in slowing, preventing, delaying or ameliorating the sequelae of diabetes, which include retinopathy, neuropathy,

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-141cardiovascular complications and nephropathy.

PTP1B, which is differentially regulated during the cell cycle (Schievella et al.. Cell. Growth Differ., 1993, 4, 239-246), is expressed in insulin sensitive tissues as two different isoforms that arise from alternate splicing of the pre-mRNA (Shifrin and Neel, J. Biol. Chem., 1993, 268, 25376-25384). The ratio of the alternatively spliced products is affected by growth factors, such as insulin, and differs in various tissues examined (Sell and Reese, Mol. Genet. Metab., 1999, 66, 189-192). In these studies the levels of the variants correlated with the plasma insulin concentration and percentage body fat. These variants may therefore be used as a biomarker for patients with chronic hyperinsulinemia or type 2 diabetes.

Definitions “Protein tyrosine phosphatase IB” is the gene product or protein of which expression is to be modulated by administration of a short antisense compound. Protein tyrosine phosphatase IB is generally referred to as PTP1B but may also be referred to as protein tyrosine phosphatase; PTPN1; RKPTP; protein tyrosine phosphatase, non-receptor type 1.

“PTP1B nucleic acid means any nucleic acid encoding PTP1B. For example, in certain embodiments, a PTP1B nucleic acid includes, without limitation, a DNA sequence encoding PTP1B, an RNA sequence transcribed from DNA encoding PTP1B, and an mRNA sequence encoding PTP1B.

“PTP1B mRNA” means an mRNA encoding a PTP1B protein.

Therapeutic indications

Antisense technology is an effective means for reducing PTP1B expression and has proven to be uniquely useful in a number of therapeutic, diagnostic, and research applications. As such, in certain embodiments, the present invention provides compounds targeted to a nucleic acid encoding PTP1B, which modulate the expression of PTP1B. Further provided herein are short antisense compounds capable of effectively inhibiting PTP IB expression.

In certain therapeutics, a subject, suspected of having a disease or disorder which can be treated by modulating the expression of PTPIB is treated by administering one or more short antisense compounds targeted to a nucleic acid encoding PTP1B. For example, in one non-limiting embodiment, the methods comprise the step of administering to an animal a therapeutically effective amount of a short antisense compound. The short antisense compounds of the present invention effectively inhibit the activity of PTPIB or inhibit the expression of PTPIB. In one embodiment, the activity or expression of PTP1B in a subject is inhibited by at least 10%, by at least 20%, by at least 25%, by at least 30%, by at least 40%, by at least 50%, by at least 60%, by at least 70%, by at least 75%, by at least 80%, by at least 85%, by at least 90%, by at least 95%, by at least 98%, by at least 99%, or by 100%. In certain embodiments, activity or expression of PTP1B in a subject is inhibited by about 30%. In certain embodiments, the activity or expression of PTPIB in a subject is inhibited by 50% or more.

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-142The reduction of the expression of PTP1B may be measured, for example, in blood, plasma, serum, adipose tissue, liver or any other body fluid, tissue or organ of the animal. Preferably, the cells contained within said fluids, tissues or organs being analyzed contain a nucleic acid molecule encoding PTP1B and/or the PTP1B protein itself.

Certain pharmaceutical and other compositions comprising the compounds of the invention are also provided. In certain embodiments short antisense compounds targeted to a PTP1B nucleic acid are utilized in pharmaceutical compositions by adding an effective amount of a compound to a suitable pharmaceutically acceptable diluent or carrier.

The short antisense compounds targeting PTP1B may have any one or more properties or characteristics of the short antisense compounds generally described herein. In certain embodiments, short antisense compounds targeting a PTP1B nucleic acid have a motif (wing - deoxy gap -wing) selected from 1-12-1,1-1-10-2,2-10-1-1, 3-10-3,2-10-3,2-10-2,1-10-1,1-10-2, 3-8-3,2-8-2,1-8-1,3-63 or 1-6-1, more preferably 1-10-1,2-10-2,3-10-3, and 1-9-2.

In certain embodiments provided herein are methods of treating an individual by administering one or more short antisense compound targeted to a PTP1B nucleic acid or a pharmaceutical composition comprising such compound. Further provided are methods of treating a subject having a disease or conditions associated with PTP1B activity by administering a short antisense compound targeted to a PTP1B nucleic acid. Diseases and conditions associated with PTP1B include but are not limited to high blood glucose or hyperglycemia, prediabetes, diabetes, Type 2 diabetes, metabolic syndrome, obesity and insulin resistance. Therefore, provided herein are methods of treating to high blood glucose or hyperglycemia, prediabetes, diabetes, Type 2 diabetes, metabolic syndrome, obesity and insulin resistance by administering a short antisense compound targeted to a PTP1B nucleic acid.

In certain embodiments the present invention provides compositions and methods for decreasing blood glucose levels in a subject or for preventing or delaying the onset of a rise in blood glucose levels in a subject, by administering to the subject a short antisense inhibitor of PTP1B expression.

In certain embodiments, the present invention provides compositions and methods for improving insulin sensitivity in a subject or for preventing or delaying the onset of insulin resistance in a subject, by administering to the subject a short antisense inhibitor of PTP1B expression.

In certain embodiments, the present invention provides compositions and methods for treating a metabolic condition in a subject or for preventing or delaying the onset of a metabolic condition in a subject, by administering to the subject a short antisense compound targeted to a PTP1B nucleic acid. Such metabolic condition may be any metabolic condition associated with PTP1B expression, including but not limited to diabetes and obesity. Also provided are methods of reducing adiposity. Also provided is a method of treating obesity wherein metabolic rate is increased.

In certain embodiments, the subject has Type 2 diabetes. In certain embodiments the subject exhibits elevated HbAlc levels In certain embodiments, HbAlc levels are at least about 6%, at least about 7%, at least about 8%, at least about 9%, at least about 10% or at least about 11%. In preferred

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-143embodiments, HbA|_C levels are reduced to about 7% or below about 7%. In certain embodiments, the subject exhibits an elevated body mass index In certain embodiments, the elevated body mass index is greater than 25 kg/m2. In certain embodiments, the subject exhibits hyperglycemia or elevated blood glucose levels. In a particular embodiment, the blood glucose levels are fasting blood glucose levels. In certain embodiments, the elevated fasting blood glucose levels are at least 130 mg/dL. In certain embodiments, the subject exhibits hyperglycemia prior to the start of treatment or exhibits fasting blood glucose levels above about 130 mg/dL, baseline HbA_k levels of at least about 7%, or body mass index of greater than 25 kg/m² or any combination thereof.

In certain embodiments a method of reducing one or more such levels by administering a short antisense compound targeted to a PTP1B nucleic acid is provided. For example, provided is a method of reducing fasting glucose levels, HbA|_C levels or, body mass index levels or any combination thereof in a subject by administering to a subject a short antisense compound targeting PTP1B. Fasting glucose may be fasting blood glucose, fasting serum glucose, or fasting plasma glucose. In some embodiments, fasting plasma glucose levels are reduced by at least about 25 mg/dL or by at least about 10 mg/dL. In a certain embodiments, said subject does not achieve normal glucose levels on a therapeutic regimen of a glucoselowering agent such as insulin, sulfonylurea, or metformin.

In certain embodiments the invention provides methods of altering lipid levels. Certain such methods reduce cholesterol, LDL and/or VLDL levels or any combination thereof in a subject by administering to the subject a short antisense compound targeted to a PTP1B nucleic acid. In certain embodiments HDL levels in a subject are increased by administering to the subject a short antisense compound targeted to a PTP1B nucleic acid. In certain embodiments, LDL: HDL ratio and/or total choIesteroliHDL ratio in a subject is reduced by administering to the subject a short antisense compound targeted to a PTP1B nucleic acid. In certain embodiments HDL: LDL ratio and/or HDL:total cholesterol ratio in a subject’s increased by administering to the subject a short antisense compound targeted to a PTP1B nucleic acid. In certain embodiments lipid profile in a subject is improved by increasing HDL, lowering LDL, lowering VLDL, lowering triglycerides, lowering apolipoprotein B levels, or lowering total cholesterol levels, or a combination thereof, by administering to the subject a short antisense compound targeted to a PTP1B nucleic acid. In such embodiments, the subject is an animal, including a human.

Combination Therapy

In certain embodiments, one or more pharmaceutical compositions comprising a short antisense compound targeted to a PTP1B nucleic acid are co-administered with one or more other pharmaceutical agents. In certain embodiments, such one or more other pharmaceutical agents are designed to treat the same disease or condition as the one or more pharmaceutical compositions of the present invention. In certain embodiments, such one or more other pharmaceutical agents are designed to treat a different disease or condition as the one or more pharmaceutical compositions of the present invention. In certain

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-144embodiments, such one or more other pharmaceutical agents are designed to treat an undesired effect of one or more pharmaceutical compositions of the present invention. In certain embodiments, one or more pharmaceutical compositions of the present invention are co-administered with another pharmaceutical agent to treat an undesired effect of that other pharmaceutical agent, in certain embodiments, one or more pharmaceutical compositions of the present invention and one or more other pharmaceutical agents are administered at the same time. In certain embodiments, one or more pharmaceutical compositions of the present invention and one or more other pharmaceutical agents are administered at different times. In certain embodiments, one or more pharmaceutical compositions of the present invention and one or more other pharmaceutical agents are prepared together in a single formulation. In certain embodiments, one or more pharmaceutical compositions of the present invention and one or more other pharmaceutical agents are prepared separately.

In certain embodiments, pharmaceutical agents that may be co-administered with a pharmaceutical composition comprising a short antisense compound targeted to a PTP1B nucleic acid include glucose-lowering agents and therapies. In some embodiments, the glucose-lowering agent is a PPAR agonist (gamma, dual, or pan), a dipeptidyl peptidase (IV) inhibitor, a GLP-1 analog, insulin or an insulin analog, an insulin secretagogue, a SGLT2 inhibitor, a human amylin analog, a biguanide, an alpha-glucosidase inhibitor, a meglitinide, a thiazolidinedione, or a sulfonylurea.

In some embodiments, the glucose-lowering therapeutic is a GLP-1 analog. In some embodiments, the GLP-1 analog is exendin-4 or liraglutide.

In other embodiments, the glucose-lowering therapeutic is a sulfonylurea. In some embodiments, the sulfonylurea is acetohexamide, chlorpropamide, tolbutamide, tolazamide, glimepiride, a glipizide, a glyburide, or a gliclazide.

In some embodiments, the glucose lowering drug is a biguanide. In some embodiments, the biguanide is metformin, and in some embodiments, blood glucose levels are decreased without increased lactic acidosis as compared to the lactic acidosis observed after treatment with metformin alone.

In some embodiments, the glucose lowering drug is a meglitinide. In some embodiments, the meglitinide is nateglinide or repaglinide.

In some embodiments, the glucose-lowering drug is a thiazolidinedione. In some embodiments, the thiazolidinedione is pioglitazone, rosiglitazone, or troglitazone. In some embodiments, blood glucose levels are decreased without greater weight gain than observed with rosiglitazone treatment alone.

In some embodiments, the glucose-lowering drug is an alpha-glucosidase inhibitor. In some embodiments, the alpha-glucosidase inhibitor is acarbose or miglitol.

In a certain embodiment, a co-administered glucose-lowering agent is ISIS 113715. In a certain embodiment, glucose-lowering therapy is therapeutic lifestyle change.

In certain such embodiments, the glucose-lowering agent is administered prior to administration of a pharmaceutical composition of the present invention. In certain such embodiments, the glucose lowering agent is administered following administration of a pharmaceutical composition of the present

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-145invention. In certain such embodiments the glucose -lowering agent is administered at the same time as a pharmaceutical composition of the present invention. In certain such embodiments the dose of a coadministered glucose -lowering agent is the same as the dose that would be administered if the glucose lowering agent was administered alone. In certain such embodiments the dose of a co-administered glucose -lowering agent is lower than the dose that would be administered if the glucose -lowering agent was administered alone. In certain such embodiments the dose of a co-administered glucose -lowering agent is greater than the dose that would be administered if the glucose -lowering agent was administered alone.

In certain embodiments, pharmaceutical agents that may be co-administered with a pharmaceutical composition comprising a short antisense compound targeted to a PTP1B nucleic acid include lipid-lowering agents. Such lipid lowering agents are discussed elsewhere in the application and are included here with respect to PTP1B. Such lipid lowering agents may be administered as described above for glucose lowering agents.

In certain embodiments, pharmaceutical agents that may be co-administered with a pharmaceutical composition comprising a short antisense compound targeted to a PTP1B nucleic acid include anti-obesity agents therapeutics. Such anti-obesity agents therapeutics may be administered as described above for glucose lowering agents.

Further provided is a method of administering a short antisense compound targeted to a PTP1B nucleic acid via injection and further including administering a topical steroid at the injection site.

Medicaments

Also provided herein are uses of a short antisense compound which is targeted to a PTP1B nucleic acid for the preparation of a medicament for reducing blood glucose levels including fasting glucose levels, and HbA|_C levels, body mass index levels or any combination thereof. The medicament can be administered during a loading period and a maintenance period. In some embodiments, the medicament is administered subcutaneously or intravenously. In other embodiments, the administration of said medicament occurs at least once daily, at least once weekly, or at least once monthly. In a particular embodiment the short antisense compound present in the medicament is administered in a dose lower than a short antisense compound with a longer sequence and particularly a sequence 20 or more nudeobases. The medicament may be administered to a subject that exhibits high blood glucose or hyperglycemia, prediabetes, diabetes, Type 2 diabetes, metabolic syndrome, obesity and insulin resistance.

Other aspects and advantages of short antisense compounds are provided herein. All aspect and advantages disclosed herein and specifically with regard to other targets is applicable with regard to compositions including short antisense compounds targeted to a PTP1B nucleic acid and methods of their use.

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-146Certain Short Antisense Compounds Targeted to a PTPIB Nucleic Acid

In certain embodiments, short antisense compounds are targeted to a PTPIB nucleic acid having the sequence of GENBANK® Accession No. NM 002827.2, incorporated herein as SEQ ID NO: 11 or the nucleotides 14178000 to 1425600 of the sequence of GENBANK® Accession No. NT_011362.9, incorporated herein as SEQ ID NO: 12. In certain such embodiments, a short antisense compound targeted to SEQ ID NO: 11 is at least 90% complementary to SEQ ID NO: 11. In certain such embodiments, a short antisense compound targeted to SEQ ID NO: 11 is at least 95% complementary to SEQ ID NO: 11. In certain such embodiments, a short antisense compound targeted to SEQ ID NO: 12 is 100% complementary to SEQ ID NO: 12. In certain such embodiments, a short antisense compound targeted to SEQ ID NO; 12 is at least 90% complementary to SEQ ID NO: 12. In certain such embodiments, a short antisense compound targeted to SEQ ID NO: 12 is at least 95% complementary to SEQ ID NO: 12. In certain such embodiments, a short antisense compound targeted to SEQ ID NO: 12 is 100% complementary to SEQ ID NO: 12.

In certain embodiments, a short antisense compound targeted to SEQ ID NO: 11 comprises a nucleotide sequence selected from the nucleotide sequences set forth in Tables 16 and 17. In certain embodiments, a short antisense compound targeted to SEQ ID NO: 12 comprises a nucleotide sequence selected from the nucleotide sequences set forth in Tables 18 and 19.

Each nucleotide sequence set forth in each Tables 16, 17, 18, and 19 is independent of any modification to a sugar moiety, an intemucleoside linkage, or a nucleobase. As such, short antisense compounds comprising a nucleotide sequence as set forth in Tables 16,17,18, and 19 may comprise, independently, one or more modifications to a sugar moiety, an intemucleoside linkage, or a nucleobase. Antisense compounds described by Isis Number (Isis NO.) indicate a combination of nucleobase sequence and one or more modifications to a sugar moiety, an intemucleoside linkage, or a nucleobase.

Tables 16 and 17 illustrate examples of short antisense compounds targeted to SEQ ID NO: 11. Table 16 illustrates short antisense compounds that are 100% complementary to SEQ ID NO: 11. Table 17 illustrates short antisense compounds that have one or two mismatches with respect to SEQ ID NO:

11. Table 18 illustrates short antisense compounds that are 100% complementary to SEQ ID NO: 12. Table 19 illustrates short antisense compounds that have 1 or 2 mismatches with respect to SEQ ID NO:

12. The column labeled ‘gapmer motif indicates the wing-gap-wing motif of each short antisense compounds. The gap segment comprises 2’-deoxynucleotides and each nucleotide of each wing segment comprises a 2’-modified sugar. The particular 2’-modified sugar is also indicated in the ‘gapmer motif column. For example, ‘2-10-2 MOE’ means a 2-10-2 gapmer motif, where a gap segment of ten 2’deoxynucleotides is flanked by wing segments of two nucleotides, where the nucleotides of the wing segments are 2’-MOE nucleotides. Intemucleoside linkages are phosphorothioate. The short antisense compounds comprise 5-methylcytidine in place of unmodified cytosine, unless “unmodified cytosine” is listed in the gapmer motif column, in which case the indicated cytosines are unmodified cytosines. For example, “5-mC in gap only” indicates that the gap segment has 5-methylcytosines, while the wing

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-147segments have unmodified cytosines.

Table 16: Short Antisense Compounds targeted to SEQ ID NO: 11

<td> ISIS NO.</td><td> 5’ Target Site</td><td> 3’ Target Site</td><td> Sequence (5’-3’)</td><td> Gapmer Motif</td><td> SEQ Π)ΝΟ</td>

<td> 147022</td><td> 177</td><td> 188</td><td> TTGTCGATCTCC</td><td> 1-10-1 MOE</td><td> 886</td>

<td> 147023</td><td> 178</td><td> 189</td><td> CTTGTCGATCTC</td><td> 1-10-1 MOE</td><td> 859</td>

<td> 147024</td><td> 179</td><td> 190</td><td> CCTTGTCGATCT</td><td> 1-10-1 MOE</td><td> 853</td>

<td> 147019</td><td> 195</td><td> 206</td><td> TCGATCTCCTCG</td><td> 1-10-1 MOE</td><td> 877</td>

<td> 147020</td><td> 196</td><td> 207</td><td> GTCGATCTCCTC</td><td> 1-10-1 MOE</td><td> 868</td>

<td> 147021</td><td> 197</td><td> 208</td><td> TGTCGATCTCCT</td><td> 1-10-1 MOE</td><td> 882</td>

<td> 147022</td><td> 198</td><td> 209</td><td> TTGTCGATCTCC</td><td> 1-10-1 MOE</td><td> 886</td>

<td> 147023</td><td> 199</td><td> 210</td><td> CTTGTCGATCTC</td><td> 1-10-1 MOE</td><td> 859</td>

<td> 147024</td><td> 200</td><td> 211</td><td> CCTTGTCGATCT</td><td> 1-10-1 MOE</td><td> 853</td>

<td> 147025</td><td> 201</td><td> 212</td><td> GCCTTGTCGATC</td><td> 1-10-1 MOE</td><td> 865</td>

<td> 147026</td><td> 202</td><td> 213</td><td> AGCCTTGTCGAT</td><td> 1-10-1 MOE</td><td> 835</td>

<td> 147027</td><td> 203</td><td> 214</td><td> CAGCCTTGTCGA</td><td> 1-10-1 MOE</td><td> 843</td>

<td> 147028</td><td> 204</td><td> 215</td><td> CCAGCCTTGTCG</td><td> 1-10-1 MOE</td><td> 846</td>

<td> 147073</td><td> 204</td><td> 215</td><td> CACTGATCCTGC</td><td> 1-10-1 MOE</td><td> 842</td>

<td> 147029</td><td> 205</td><td> 216</td><td> CCCAGCCTTGTC</td><td> 1-10-1 MOE</td><td> 848</td>

<td> 147030</td><td> 206</td><td> 217</td><td> TCCCAGCCTTGT</td><td> 1-10-1 MOE</td><td> 874</td>

<td> 147036</td><td> 212</td><td> 223</td><td> CCCAGTTCCCAG</td><td> 1-10-1 MOE</td><td> 849</td>

<td> 147037</td><td> 213</td><td> 224</td><td> GCCCAGTTCCCA</td><td> 1-10-1 MOE</td><td> 863</td>

<td> 147038</td><td> 214</td><td> 225</td><td> CGCCCAGTTCCC</td><td> 1-10-1 MOE</td><td> 855</td>

<td> 147039</td><td> 215</td><td> 226</td><td> CCGCCCAGTTCC</td><td> 1-10-1 MOE</td><td> 850</td>

<td> 147040</td><td> 216</td><td> 227</td><td> GCCGCCCAGTTC</td><td> 1-10-1 MOE</td><td> 864</td>

<td> 147041</td><td> 217</td><td> 228</td><td> AGCCGCCCAGTT</td><td> 1-10-1 MOE</td><td> 834</td>

<td> 147073</td><td> 311</td><td> 322</td><td> CACTGATCCTGC</td><td> 1-10-1 MOE</td><td> 842</td>

<td> 147042</td><td> 323</td><td> 334</td><td> GGTCAAAAGGGC</td><td> 1-10-1 MOE</td><td> 866</td>

<td> 147043</td><td> 324</td><td> 335</td><td> TGGTCAAAAGGG</td><td> 1-10-1 MOE</td><td> 881</td>

<td> 147044</td><td> 325</td><td> 336</td><td> GTGGTCAAAAGG</td><td> 1-10-1 MOE</td><td> 869</td>

<td> 147045</td><td> 326</td><td> 337</td><td> TGTGGTCAAAAG</td><td> 1-10-1 MOE</td><td> 883</td>

<td> 147046</td><td> 327</td><td> 338</td><td> CTGTGGTCAAAA</td><td> 1-10-1 MOE</td><td> 858</td>

<td> 147047</td><td> 328</td><td> 339</td><td> ACTGTGGTCAAA</td><td> 1-10-1 MOE</td><td> 833</td>

<td> 147051</td><td> 332</td><td> 343</td><td> TCCGACTGTGGT</td><td> 1-10-1 MOE</td><td> 875</td>

<td> 147052</td><td> 333</td><td> 344</td><td> ATCCGACTGTGG</td><td> 1-10-1 MOE</td><td> 837</td>

<td> 147053</td><td> 334</td><td> 345</td><td> AATCCGACTGTG</td><td> 1-10-1 MOE</td><td> 829</td>

<td> 147054</td><td> 335</td><td> 346</td><td> TAATCCGACTGT</td><td> 1-10-1 MOE</td><td> 871</td>

<td> 147055</td><td> 336</td><td> 347</td><td> TTAATCCGACTG</td><td> 1-10-1 MOE</td><td> 884</td>

<td> 147056</td><td> 337</td><td> 348</td><td> TTTAATCCGACT</td><td> 1-10-1 MOE</td><td> 887</td>

<td> 147057</td><td> 338</td><td> 349</td><td> ATTTAATCCGAC</td><td> 1-10-1 MOE</td><td> 839</td>

<td> 147058</td><td> 339</td><td> 350</td><td> AATTTAATCCGA</td><td> 1-10-1 MOE</td><td> 830</td>

<td> 147059</td><td> 340</td><td> 351</td><td> CAATTTAATCCG</td><td> 1-10-1 MOE</td><td> 840</td>

<td> 147060</td><td> 341</td><td> 352</td><td> GCAATTTAATCC</td><td> 1-10-1 MOE</td><td> 861</td>

<td> 147061</td><td> 342</td><td> 353</td><td> TGCAATTTAATC</td><td> 1-10-1 MOE</td><td> 879</td>

<td> 147045</td><td> 679</td><td> 690</td><td> TGTGGTCAAAAG</td><td> 1-10-1 MOE</td><td> 883</td>

<td> 147046</td><td> 680</td><td> 691</td><td> CTGTGGTCAAAA</td><td> 1-10-1 MOE</td><td> 858</td>

<td> 147045</td><td> 787</td><td> 798</td><td> TGTGGTCAAAAG</td><td> 1-10-1 MOE</td><td> 883</td>

<td> 147046</td><td> 788</td><td> 799</td><td> CTGTGGTCAAAA</td><td> 1-10-1 MOE</td><td> 858</td>

<td> 147066</td><td> 816</td><td> 827</td><td> CCTGCACTGACG</td><td> 1-10-1 MOE</td><td> 851</td>

<td> 404131</td><td> 992</td><td> 1005</td><td> ACCTTCGATCACAG</td><td> 2-10-2 MOE</td><td> 831</td>

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<td> 147062</td><td> 1024</td><td> 1035</td><td> CACTGACGAGTC</td><td> 1-10-1 MOE</td><td> 841</td>

<td> 147063</td><td> 1025</td><td> 1036</td><td> GCACTGACGAGT</td><td> 1-10-1 MOE</td><td> 862</td>

<td> 147064</td><td> 1026</td><td> 1037</td><td> TGCACTGACGAG</td><td> 1-10-1 MOE</td><td> 880</td>

<td> 147065</td><td> 1027</td><td> 1038</td><td> CTGCACTGACGA</td><td> 1-10-1 MOE</td><td> 857</td>

<td> 147066</td><td> 1028</td><td> 1039</td><td> CCTGCACTGACG</td><td> 1-10-1 MOE</td><td> 851</td>

<td> 147067</td><td> 1029</td><td> 1040</td><td> TCCTGCACTGAC</td><td> 1-10-1 MOE</td><td> 876</td>

<td> 147068</td><td> 1030</td><td> 1041</td><td> ATCCTGCACTGA</td><td> 1-10-1 MOE</td><td> 838</td>

<td> 147069</td><td> 1031</td><td> 1042</td><td> GATCCTGCACTG</td><td> 1-10-1 MOE</td><td> 860</td>

<td> 147070</td><td> 1032</td><td> 1043</td><td> TGATCCTGCACT</td><td> 1-10-1 MOE</td><td> 878</td>

<td> 147071</td><td> 1033</td><td> 1044</td><td> CTGATCCTGCAC</td><td> 1-10-1 MOE</td><td> 856</td>

<td> 147072</td><td> 1034</td><td> 1045</td><td> ACTGATCCTGCA</td><td> 1-10-1 MOE</td><td> 832</td>

<td> 147073</td><td> 1035</td><td> 1046</td><td> CACTGATCCTGC</td><td> 1-10-1 MOE</td><td> 842</td>

<td> 147067</td><td> 1199</td><td> 1210</td><td> TCCTGCACTGAC</td><td> 1-10-1 MOE</td><td> 876</td>

<td> 147040</td><td> 1288</td><td> 1299</td><td> GCCGCCCAGTTC</td><td> 1-10-1 MOE</td><td> 864</td>

<td> 147040</td><td> 1396</td><td> 1407</td><td> GCCGCCCAGTTC</td><td> 1-10-1 MOE</td><td> 864</td>

<td> 147022</td><td> 1868</td><td> 1879</td><td> TTGTCGATCTCC</td><td> 1-10-1 MOE</td><td> 886</td>

<td> 147023</td><td> 1869</td><td> 1880</td><td> CTTGTCGATCTC</td><td> 1-10-1 MOE</td><td> 859</td>

<td> 147024</td><td> 1870</td><td> 1881</td><td> CCTTGTCGATCT</td><td> 1-10-1 MOE</td><td> 853</td>

<td> 147019</td><td> 1886</td><td> 1897</td><td> TCGATCTCCTCG</td><td> 1-10-1 MOE</td><td> 877</td>

<td> 147020</td><td> 1887</td><td> 1898</td><td> GTCGATCTCCTC</td><td> 1-10-1 MOE</td><td> 868</td>

<td> 147021</td><td> 1888</td><td> 1899</td><td> TGTCGATCTCCT</td><td> 1-10-1 MOE</td><td> 882</td>

<td> 147022</td><td> 1889</td><td> 1900</td><td> TTGTCGATCTCC</td><td> 1-10-1 MOE</td><td> 886</td>

<td> 147023</td><td> 1890</td><td> 1901</td><td> CTTGTCGATCTC</td><td> 1-10-1 MOE</td><td> 859</td>

<td> 147025</td><td> 1892</td><td> 1903</td><td> GCCTTGTCGATC</td><td> 1-10-1 MOE</td><td> 865</td>

<td> 147027</td><td> 1894</td><td> 1905</td><td> CAGCCTTGTCGA</td><td> 1-10-1 MOE</td><td> 843</td>

<td> 147028</td><td> 1895</td><td> 1906</td><td> CCAGCCTTGTCG</td><td> 1-10-1 MOE</td><td> 846</td>

<td> 147030</td><td> 1897</td><td> 1908</td><td> TCCCAGCCTTGT</td><td> 1-10-1 MOE</td><td> 874</td>

<td> 147037</td><td> 1904</td><td> 1915</td><td> GCCCAGTTCCCA</td><td> 1-10-1 MOE</td><td> 863</td>

<td> 147038</td><td> 1905</td><td> 1916</td><td> CGCCCAGTTCCC</td><td> 1-10-1 MOE</td><td> 855</td>

<td> 147040</td><td> 1907</td><td> 1918</td><td> GCCGCCCAGTTC</td><td> 1-10-1 MOE</td><td> 864</td>

<td> 147041</td><td> 1908</td><td> 1919</td><td> AGCCGCCCAGTT</td><td> 1-10-1 MOE</td><td> 834</td>

<td> 147022</td><td> 1976</td><td> 1987</td><td> TTGTCGATCTCC</td><td> 1-10-1 MOE</td><td> 886</td>

<td> 147023</td><td> 1977</td><td> 1988</td><td> CTTGTCGATCTC</td><td> 1-10-1 MOE</td><td> 859</td>

<td> 147024</td><td> 1978</td><td> 1989</td><td> CCTTGTCGATCT</td><td> 1-10-1 MOE</td><td> 853</td>

<td> 147020</td><td> 1995</td><td> 2006</td><td> GTCGATCTCCTC</td><td> 1-10-1 MOE</td><td> 868</td>

<td> 147021</td><td> 1996</td><td> 2007</td><td> TGTCGATCTCCT</td><td> 1-10-1 MOE</td><td> 882</td>

<td> 147022</td><td> 1997</td><td> 2008</td><td> TTGTCGATCTCC</td><td> 1-10-1 MOE</td><td> 886</td>

<td> 147023</td><td> 1998</td><td> 2009</td><td> CTTGTCGATCTC</td><td> 1-10-1 MOE</td><td> 859</td>

<td> 147024</td><td> 1999</td><td> 2010</td><td> CCTTGTCGATCT</td><td> 1-10-1 MOE</td><td> 853</td>

<td> 147025</td><td> 2000</td><td> 2011</td><td> GCCTTGTCGATC</td><td> 1-10-1 MOE</td><td> 865</td>

<td> 147026</td><td> 2001</td><td> 2012</td><td> AGCCTTGTCGAT</td><td> 1-10-1 MOE</td><td> 835</td>

<td> 147027</td><td> 2002</td><td> 2013</td><td> CAGCCTTGTCGA</td><td> 1-10-1 MOE</td><td> 843</td>

<td> 147028</td><td> 2003</td><td> 2014</td><td> CCAGCCTTGTCG</td><td> 1-10-1 MOE</td><td> 846</td>

<td> 147029</td><td> 2004</td><td> 2015</td><td> CCCAGCCTTGTC</td><td> 1-10-1 MOE</td><td> 848</td>

<td> 147030</td><td> 2005</td><td> 2016</td><td> TCCCAGCCTTGT</td><td> 1-10-1 MOE</td><td> 874</td>

<td> 147036</td><td> 2011</td><td> 2022</td><td> CCCAGTTCCCAG</td><td> 1-10-1 MOE</td><td> 849</td>

<td> 147037</td><td> 2012</td><td> 2023</td><td> GCCCAGTTCCCA</td><td> 1-10-1 MOE</td><td> 863</td>

<td> 147038</td><td> 2013</td><td> 2024</td><td> CGCCCAGTTCCC</td><td> 1-10-1 MOE</td><td> 855</td>

<td> 147039</td><td> 2014</td><td> 2025</td><td> CCGCCCAGTTCC</td><td> 1-10-1 MOE</td><td> 850</td>

<td> 147040</td><td> 2015</td><td> 2026</td><td> GCCGCCCAGTTC</td><td> 1-10-1 MOE</td><td> 864</td>

<td> 147041</td><td> 2016</td><td> 2027</td><td> AGCCGCCCAGTT</td><td> 1-10-1 MOE</td><td> 834</td>

<td> 404199</td><td> 2366</td><td> 2379</td><td> GGTCATGCACAGGC</td><td> 2-10-2 MOE</td><td> 867</td>

<td> 404134</td><td> 2369</td><td> 2382</td><td> TCAGGTCATGCACA</td><td> 2-10-2 MOE</td><td> 873</td>

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<td> 404132</td><td> 2548</td><td> 2561</td><td> CCTTGGAATGTCTG</td><td> 2-10-2 MOE</td><td> 852</td>

<td> 147020</td><td> 2613</td><td> 2624</td><td> GTCGATCTCCTC</td><td> 1-10-1 MOE</td><td> 868</td>

<td> 147020</td><td> 2721</td><td> 2732</td><td> GTCGATCTCCTC</td><td> 1-10-1 MOE</td><td> 868</td>

<td> 404133</td><td> 3289</td><td> 3302</td><td> TATTCCATGGCCAT</td><td> 2-10-2 MOE</td><td> 872</td>

<td> 147032</td><td> 6220</td><td> 6231</td><td> GTTCCCAGCCTT</td><td> 1-10-1 MOE</td><td> 870</td>

<td> 147033</td><td> 6221</td><td> 6232</td><td> AGTTCCCAGCCT</td><td> 1-10-1 MOE</td><td> 836</td>

<td> 147034</td><td> 6222</td><td> 6233</td><td> CAGTTCCCAGCC</td><td> 1-10-1 MOE</td><td> 844</td>

<td> 147044</td><td> 6288</td><td> 6299</td><td> GTGGTCAAAAGG</td><td> 1-10-1 MOE</td><td> 869</td>

<td> 147045</td><td> 6289</td><td> 6300</td><td> TGTGGTCAAAAG</td><td> 1-10-1 MOE</td><td> 883</td>

<td> 147032</td><td> 6329</td><td> 6340</td><td> GTTCCCAGCCTT</td><td> 1-10-1 MOE</td><td> 870</td>

<td> 147033</td><td> 6330</td><td> 6341</td><td> AGTTCCCAGCCT</td><td> 1-10-1 MOE</td><td> 836</td>

<td> 147034</td><td> 6331</td><td> 6342</td><td> CAGTTCCCAGCC</td><td> 1-10-1 MOE</td><td> 844</td>

<td> 147044</td><td> 6397</td><td> 6408</td><td> GTGGTCAAAAGG</td><td> 1-10-1 MOE</td><td> 869</td>

<td> 147045</td><td> 6398</td><td> 6409</td><td> TGTGGTCAAAAG</td><td> 1-10-1 MOE</td><td> 883</td>

<td> 147058</td><td> 7057</td><td> 7068</td><td> AATTTAATCCGA</td><td> 1-10-1 MOE</td><td> 830</td>

<td> 147059</td><td> 7058</td><td> 7069</td><td> CAATTTAATCCG</td><td> 1-10-1 MOE</td><td> 840</td>

<td> 147060</td><td> 7059</td><td> 7070</td><td> GCAATTTAATCC</td><td> 1-10-1 MOE</td><td> 861</td>

<td> 147058</td><td> 7166</td><td> 7177</td><td> AATTTAATCCGA</td><td> 1-10-1 MOE</td><td> 830</td>

<td> 147059</td><td> 7167</td><td> 7178</td><td> CAATTTAATCCG</td><td> 1-10-1 MOE</td><td> 840</td>

<td> 147041</td><td> 8084</td><td> 8095</td><td> AGCCGCCCAGTT</td><td> 1-10-1 MOE</td><td> 834</td>

<td> 147041</td><td> 8192</td><td> 8203</td><td> AGCCGCCCAGTT</td><td> 1-10-1 MOE</td><td> 834</td>

<td> 147027</td><td> 8630</td><td> 8641</td><td> CAGCCTTGTCGA</td><td> 1-10-1 MOE</td><td> 843</td>

<td> 147028</td><td> 8631</td><td> 8642</td><td> CCAGCCTTGTCG</td><td> 1-10-1 MOE</td><td> 846</td>

<td> 147027</td><td> 8738</td><td> 8749</td><td> CAGCCTTGTCGA</td><td> 1-10-1 MOE</td><td> 843</td>

<td> 147028</td><td> 8739</td><td> 8750</td><td> CCAGCCTTGTCG</td><td> 1-10-1 MOE</td><td> 846</td>

<td> 147043</td><td> 10957</td><td> 10968</td><td> TGGTCAAAAGGG</td><td> 1-10-1 MOE</td><td> 881</td>

<td> 147044</td><td> 10958</td><td> 10969</td><td> GTGGTCAAAAGG</td><td> 1-10-1 MOE</td><td> 869</td>

<td> 147043</td><td> 11065</td><td> 11076</td><td> TGGTCAAAAGGG</td><td> 1-10-1 MOE</td><td> 881</td>

<td> 147044</td><td> 11066</td><td> 11077</td><td> GTGGTCAAAAGG</td><td> 1-10-1 MOE</td><td> 869</td>

<td> 147071</td><td> 11605</td><td> 11616</td><td> CTGATCCTGCAC</td><td> 1-10-1 MOE</td><td> 856</td>

<td> 147070</td><td> 11611</td><td> 11622</td><td> TGATCCTGCACT</td><td> 1-10-1 MOE</td><td> 878</td>

<td> 147071</td><td> 11612</td><td> 11623</td><td> CTGATCCTGCAC</td><td> 1-10-1 MOE</td><td> 856</td>

<td> 147072</td><td> 12294</td><td> 12305</td><td> ACTGATCCTGCA</td><td> 1-10-1 MOE</td><td> 832</td>

<td> 147072</td><td> 12299</td><td> 12310</td><td> ACTGATCCTGCA</td><td> 1-10-1 MOE</td><td> 832</td>

<td> 147030</td><td> 12805</td><td> 12816</td><td> TCCCAGCCTTGT</td><td> 1-10-1 MOE</td><td> 874</td>

<td> 147031</td><td> 12806</td><td> 12817</td><td> TTCCCAGCCTTG</td><td> 1-10-1 MOE</td><td> 885</td>

<td> 147053</td><td> 12939</td><td> 12950</td><td> AATCCGACTGTG</td><td> 1-10-1 MOE</td><td> 829</td>

<td> 147030</td><td> 12986</td><td> 12997</td><td> TCCCAGCCTTGT</td><td> 1-10-1 MOE</td><td> 874</td>

<td> 147031</td><td> 12987</td><td> 12998</td><td> TTCCCAGCCTTG</td><td> 1-10-1 MOE</td><td> 885</td>

<td> 147053</td><td> 13120</td><td> 13131</td><td> AATCCGACTGTG</td><td> 1-10-1 MOE</td><td> 829</td>

<td> 147051</td><td> 13162</td><td> 13173</td><td> TCCGACTGTGGT</td><td> 1-10-1 MOE</td><td> 875</td>

<td> 147061</td><td> 13316</td><td> 13327</td><td> TGCAATTTAATC</td><td> 1-10-1 MOE</td><td> 879</td>

<td> 147047</td><td> 13339</td><td> 13350</td><td> ACTGTGGTCAAA</td><td> 1-10-1 MOE</td><td> 833</td>

<td> 147029</td><td> 14058</td><td> 14069</td><td> CCCAGCCTTGTC</td><td> 1-10-1 MOE</td><td> 848</td>

<td> 147029</td><td> 14239</td><td> 14250</td><td> CCCAGCCTTGTC</td><td> 1-10-1 MOE</td><td> 848</td>

<td> 147067</td><td> 15560</td><td> 15571</td><td> TCCTGCACTGAC</td><td> 1-10-1 MOE</td><td> 876</td>

<td> 147068</td><td> 15561</td><td> 15572</td><td> ATCCTGCACTGA</td><td> 1-10-1 MOE</td><td> 838</td>

<td> 147067</td><td> 15742</td><td> 15753</td><td> TCCTGCACTGAC</td><td> 1-10-1 MOE</td><td> 876</td>

<td> 147069</td><td> 15744</td><td> 15755</td><td> GATCCTGCACTG</td><td> 1-10-1 MOE</td><td> 860</td>

<td> 147042</td><td> 16561</td><td> 16572</td><td> GGTCAAAAGGGC</td><td> 1-10-1 MOE</td><td> 866</td>

<td> 147042</td><td> 16727</td><td> 16738</td><td> GGTCAAAAGGGC</td><td> 1-10-1 MOE</td><td> 866</td>

<td> 147030</td><td> 17619</td><td> 17630</td><td> TCCCAGCCTTGT</td><td> 1-10-1 MOE</td><td> 874</td>

<td> 147064</td><td> 17762</td><td> 17773</td><td> TGCACTGACGAG</td><td> 1-10-1 MOE</td><td> 880</td>

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<td> 147030</td><td> 17787</td><td> 17798</td><td> TCCCAGCCTTGT</td><td> 1-10-1 MOE</td><td> 874</td>

<td> 147064</td><td> 17930</td><td> 17941</td><td> TGCACTGACGAG</td><td> 1-10-1 MOE</td><td> 880</td>

<td> 147042</td><td> 19201</td><td> 19212</td><td> GGTCAAAAGGGC</td><td> 1-10-1 MOE</td><td> 866</td>

<td> 147042</td><td> 19369</td><td> 19380</td><td> GGTCAAAAGGGC</td><td> 1-10-1 MOE</td><td> 866</td>

<td> 147027</td><td> 21190</td><td> 21201</td><td> CAGCCTTGTCGA</td><td> 1-10-1 MOE</td><td> 843</td>

<td> 147028</td><td> 21191</td><td> 21202</td><td> CCAGCCTTGTCG</td><td> 1-10-1 MOE</td><td> 846</td>

<td> 147027</td><td> 21358</td><td> 21369</td><td> CAGCCTTGTCGA</td><td> 1-10-1 MOE</td><td> 843</td>

<td> 147028</td><td> 21359</td><td> 21370</td><td> CCAGCCTTGTCG</td><td> 1-10-1 MOE</td><td> 846</td>

<td> 147070</td><td> 22021</td><td> 22032</td><td> TGATCCTGCACT</td><td> 1-10-1 MOE</td><td> 878</td>

<td> 147070</td><td> 22189</td><td> 22200</td><td> TGATCCTGCACT</td><td> 1-10-1 MOE</td><td> 878</td>

<td> 147047</td><td> 22606</td><td> 22617</td><td> ACTGTGGTCAAA</td><td> 1-10-1 MOE</td><td> 833</td>

<td> 147043</td><td> 24318</td><td> 24329</td><td> TGGTCAAAAGGG</td><td> 1-10-1 MOE</td><td> 881</td>

<td> 147044</td><td> 24319</td><td> 24330</td><td> GTGGTCAAAAGG</td><td> 1-10-1 MOE</td><td> 869</td>

<td> 147045</td><td> 24320</td><td> 24331</td><td> TGTGGTCAAAAG</td><td> 1-10-1 MOE</td><td> 883</td>

<td> 147046</td><td> 24321</td><td> 24332</td><td> CTGTGGTCAAAA</td><td> 1-10-1 MOE</td><td> 858</td>

<td> 147043</td><td> 24486</td><td> 24497</td><td> TGGTCAAAAGGG</td><td> 1-10-1 MOE</td><td> 881</td>

<td> 147044</td><td> 24487</td><td> 24498</td><td> GTGGTCAAAAGG</td><td> 1-10-1 MOE</td><td> 869</td>

<td> 147046</td><td> 24489</td><td> 24500</td><td> CTGTGGTCAAAA</td><td> 1-10-1 MOE</td><td> 858</td>

<td> 147047</td><td> 24490</td><td> 24501</td><td> ACTGTGGTCAAA</td><td> 1-10-1 MOE</td><td> 833</td>

<td> 147040</td><td> 25065</td><td> 25076</td><td> GCCGCCCAGTTC</td><td> 1-10-1 MOE</td><td> 864</td>

<td> 147041</td><td> 25066</td><td> 25077</td><td> AGCCGCCCAGTT</td><td> 1-10-1 MOE</td><td> 834</td>

<td> 147046</td><td> 25160</td><td> 25171</td><td> CTGTGGTCAAAA</td><td> 1-10-1 MOE</td><td> 858</td>

<td> 147039</td><td> 25232</td><td> 25243</td><td> CCGCCCAGTTCC</td><td> 1-10-1 MOE</td><td> 850</td>

<td> 147040</td><td> 25233</td><td> 25244</td><td> GCCGCCCAGTTC</td><td> 1-10-1 MOE</td><td> 864</td>

<td> 147041</td><td> 25234</td><td> 25245</td><td> AGCCGCCCAGTT</td><td> 1-10-1 MOE</td><td> 834</td>

<td> 147046</td><td> 25328</td><td> 25339</td><td> CTGTGGTCAAAA</td><td> 1-10-1 MOE</td><td> 858</td>

<td> 147057</td><td> 25508</td><td> 25519</td><td> ATTTAATCCGAC</td><td> 1-10-1 MOE</td><td> 839</td>

<td> 147061</td><td> 25512</td><td> 25523</td><td> TGCAATTTAATC</td><td> 1-10-1 MOE</td><td> 879</td>

<td> 147057</td><td> 25676</td><td> 25687</td><td> ATTTAATCCGAC</td><td> 1-10-1 MOE</td><td> 839</td>

<td> 147069</td><td> 28878</td><td> 28889</td><td> GATCCTGCACTG</td><td> 1-10-1 MOE</td><td> 860</td>

<td> 147070</td><td> 28879</td><td> 28890</td><td> TGATCCTGCACT</td><td> 1-10-1 MOE</td><td> 878</td>

<td> 147053</td><td> 30133</td><td> 30144</td><td> AATCCGACTGTG</td><td> 1-10-1 MOE</td><td> 829</td>

<td> 147053</td><td> 30278</td><td> 30289</td><td> AATCCGACTGTG</td><td> 1-10-1 MOE</td><td> 829</td>

<td> 147054</td><td> 30864</td><td> 30875</td><td> TAATCCGACTGT</td><td> 1-10-1 MOE</td><td> 871</td>

<td> 147043</td><td> 30985</td><td> 30996</td><td> TGGTCAAAAGGG</td><td> 1-10-1 MOE</td><td> 881</td>

<td> 147054</td><td> 31011</td><td> 31022</td><td> TAATCCGACTGT</td><td> 1-10-1 MOE</td><td> 871</td>

<td> 147043</td><td> 31133</td><td> 31144</td><td> TGGTCAAAAGGG</td><td> 1-10-1 MOE</td><td> 881</td>

<td> 147036</td><td> 32233</td><td> 32244</td><td> CCCAGTTCCCAG</td><td> 1-10-1 MOE</td><td> 849</td>

<td> 147072</td><td> 32372</td><td> 32383</td><td> ACTGATCCTGCA</td><td> 1-10-1 MOE</td><td> 832</td>

<td> 147072</td><td> 32520</td><td> 32531</td><td> ACTGATCCTGCA</td><td> 1-10-1 MOE</td><td> 832</td>

<td> 147069</td><td> 33056</td><td> 33067</td><td> GATCCTGCACTG</td><td> 1-10-1 MOE</td><td> 860</td>

<td> 147070</td><td> 33057</td><td> 33068</td><td> TGATCCTGCACT</td><td> 1-10-1 MOE</td><td> 878</td>

<td> 147071</td><td> 33058</td><td> 33069</td><td> CTGATCCTGCAC</td><td> 1-10-1 MOE</td><td> 856</td>

<td> 147051</td><td> 33126</td><td> 33137</td><td> TCCGACTGTGGT</td><td> 1-10-1 MOE</td><td> 875</td>

<td> 147070</td><td> 33205</td><td> 33216</td><td> TGATCCTGCACT</td><td> 1-10-1 MOE</td><td> 878</td>

<td> 147071</td><td> 33206</td><td> 33217</td><td> CTGATCCTGCAC</td><td> 1-10-1 MOE</td><td> 856</td>

<td> 147051</td><td> 33274</td><td> 33285</td><td> TCCGACTGTGGT</td><td> 1-10-1 MOE</td><td> 875</td>

<td> 147046</td><td> 33318</td><td> 33329</td><td> CTGTGGTCAAAA</td><td> 1-10-1 MOE</td><td> 858</td>

<td> 147049</td><td> 33321</td><td> 33332</td><td> CGACTGTGGTCA</td><td> 1-10-1 MOE</td><td> 854</td>

<td> 147051</td><td> 33323</td><td> 33334</td><td> TCCGACTGTGGT</td><td> 1-ib-l MOE</td><td> 875</td>

<td> 147046</td><td> 33466</td><td> 33477</td><td> CTGTGGTCAAAA</td><td> 1-10-1 MOE</td><td> 858</td>

<td> 147047</td><td> 33467</td><td> 33478</td><td> ACTGTGGTCAAA</td><td> 1-10-1 MOE</td><td> 833</td>

<td> 147051</td><td> 33471</td><td> 33482</td><td> TCCGACTGTGGT</td><td> 1-10-1 MOE</td><td> 875</td>

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<td> 147046</td><td> 33640</td><td> 33651</td><td> CTGTGGTCAAAA</td><td> 1-10-1 MOE</td><td> 858</td>

<td> 147051</td><td> 33645</td><td> 33656</td><td> TCCGACTGTGGT</td><td> 1-10-1 MOE</td><td> 875</td>

<td> 147046</td><td> 33788</td><td> 33799</td><td> CTGTGGTCAAAA</td><td> 1-10-1 MOE</td><td> 858</td>

<td> 147051</td><td> 33793</td><td> 33804</td><td> TCCGACTGTGGT</td><td> 1-10-1 MOE</td><td> 875</td>

<td> 147059</td><td> 35437</td><td> 35448</td><td> CAATTTAATCCG</td><td> 1-10-1 MOE</td><td> 840</td>

<td> 147060</td><td> 35438</td><td> 35449</td><td> GCAATTTAATCC</td><td> 1-10-1 MOE</td><td> 861</td>

<td> 147060</td><td> 35586</td><td> 35597</td><td> GCAATTTAATCC</td><td> 1-10-1 MOE</td><td> 861</td>

<td> 147021</td><td> 36093</td><td> 36104</td><td> TGTCGATCTCCT</td><td> 1-10-1 MOE</td><td> 882</td>

<td> 147061</td><td> 36250</td><td> 36261</td><td> TGCAATTTAATC</td><td> 1-10-1 MOE</td><td> 879</td>

<td> 147061</td><td> 36398</td><td> 36409</td><td> TGCAATTTAATC</td><td> 1-10-1 MOE</td><td> 879</td>

<td> 147073</td><td> 37485</td><td> 37496</td><td> CACTGATCCTGC</td><td> 1-10-1 MOE</td><td> 842</td>

<td> 147073</td><td> 37633</td><td> 37644</td><td> CACTGATCCTGC</td><td> 1-10-1 MOE</td><td> 842</td>

<td> 147043</td><td> 40214</td><td> 40225</td><td> TGGTCAAAAGGG</td><td> 1-10-1 MOE</td><td> 881</td>

<td> 147061</td><td> 40353</td><td> 40364</td><td> TGCAATTTAATC</td><td> 1-10-1 MOE</td><td> 879</td>

<td> 147043</td><td> 40362</td><td> 40373</td><td> TGGTCAAAAGGG</td><td> 1-10-1 MOE</td><td> 881</td>

<td> 147061</td><td> 40501</td><td> 40512</td><td> TGCAATTTAATC</td><td> 1-10-1 MOE</td><td> 879</td>

<td> 147031</td><td> 42527</td><td> 42538</td><td> TTCCCAGCCTTG</td><td> 1-10-1 MOE</td><td> 885</td>

<td> 147032</td><td> 42528</td><td> 42539</td><td> GTTCCCAGCCTT</td><td> 1-10-1 MOE</td><td> 870</td>

<td> 147034</td><td> 42530</td><td> 42541</td><td> CAGTTCCCAGCC</td><td> 1-10-1 MOE</td><td> 844</td>

<td> 147031</td><td> 42675</td><td> 42686</td><td> TTCCCAGCCTTG</td><td> 1-10-1 MOE</td><td> 885</td>

<td> 147032</td><td> 42676</td><td> 42687</td><td> GTTCCCAGCCTT</td><td> 1-10-1 MOE</td><td> 870</td>

<td> 147033</td><td> 42677</td><td> 42688</td><td> AGTTCCCAGCCT</td><td> 1-10-1 MOE</td><td> 836</td>

<td> 147034</td><td> 42678</td><td> 42689</td><td> CAGTTCCCAGCC</td><td> 1-10-1 MOE</td><td> 844</td>

<td> 147074</td><td> 43848</td><td> 43859</td><td> CCACTGATCCTG</td><td> 1-10-1 MOE</td><td> 845</td>

<td> 147074</td><td> 43996</td><td> 44007</td><td> CCACTGATCCTG</td><td> 1-10-1 MOE</td><td> 845</td>

<td> 147051</td><td> 45402</td><td> 45413</td><td> TCCGACTGTGGT</td><td> 1-10-1 MOE</td><td> 875</td>

<td> 147051</td><td> 45550</td><td> 45561</td><td> TCCGACTGTGGT</td><td> 1-10-1 MOE</td><td> 875</td>

<td> 147074</td><td> 46125</td><td> 46136</td><td> CCACTGATCCTG</td><td> 1-10-1 MOE</td><td> 845</td>

<td> 147057</td><td> 46313</td><td> 46324</td><td> ATTTAATCCGAC</td><td> 1-10-1 MOE</td><td> 839</td>

<td> 147058</td><td> 46314</td><td> 46325</td><td> AATTTAATCCGA</td><td> 1-10-1 MOE</td><td> 830</td>

<td> 147059</td><td> Π46315</td><td> 46326</td><td> CAATTTAATCCG</td><td> 1-10-1 MOE</td><td> 840</td>

<td> 147061</td><td> 46317</td><td> 46328</td><td> TGCAATTTAATC</td><td> 1-10-1 MOE</td><td> 879</td>

<td> 147057</td><td> 46461</td><td> 46472</td><td> ATTTAATCCGAC</td><td> 1-10-1 MOE</td><td> 839</td>

<td> 147059</td><td> 46463</td><td> 46474</td><td> CAATTTAATCCG</td><td> 1-10-1 MOE</td><td> 840</td>

<td> 147061</td><td> 46465</td><td> 46476</td><td> TGCAATTTAATC</td><td> 1-10-1 MOE</td><td> 879</td>

<td> 147058</td><td> 47413</td><td> 47424</td><td> AATTTAATCCGA</td><td> 1-10-1 MOE</td><td> 830</td>

<td> 147073</td><td> 48221</td><td> 48232</td><td> CACTGATCCTGC</td><td> 1-10-1 MOE</td><td> 842</td>

<td> 147073</td><td> 48369</td><td> 48380</td><td> CACTGATCCTGC</td><td> 1-10-1 MOE</td><td> 842</td>

<td> 147074</td><td> 48370</td><td> 48381</td><td> CCACTGATCCTG</td><td> 1-10-1 MOE</td><td> 845</td>

<td> 147027</td><td> 48566</td><td> 48577</td><td> CAGCCTTGTCGA</td><td> 1-10-1 MOE</td><td> 843</td>

<td> 147027</td><td> 48714</td><td> 48725</td><td> CAGCCTTGTCGA</td><td> 1-10-1 MOE</td><td> 843</td>

<td> 147028</td><td> 48715</td><td> 48726</td><td> CCAGCCTTGTCG</td><td> 1-10-1 MOE</td><td> 846</td>

<td> 147067</td><td> 49050</td><td> 49061</td><td> TCCTGCACTGAC</td><td> 1-10-1 MOE</td><td> 876</td>

<td> 147068</td><td> 49051</td><td> 49062</td><td> ATCCTGCACTGA</td><td> 1-10-1 MOE</td><td> 838</td>

<td> 147067</td><td> 49198</td><td> 49209</td><td> TCCTGCACTGAC</td><td> 1-10-1 MOE</td><td> 876</td>

<td> 147073</td><td> 49524</td><td> 49535</td><td> CACTGATCCTGC</td><td> 1-10-1 MOE</td><td> 842</td>

<td> 147073</td><td> 49672</td><td> 49683</td><td> CACTGATCCTGC</td><td> 1-10-1 MOE</td><td> 842</td>

<td> 147074</td><td> 49673</td><td> 49684</td><td> CCACTGATCCTG</td><td> 1-10-1 MOE</td><td> 845</td>

<td> 147036</td><td> 50421</td><td> 50432</td><td> CCCAGTTCCCAG</td><td> 1-10-1 MOE</td><td> 849</td>

<td> 147036</td><td> 52292</td><td> 52303</td><td> CCCAGTTCCCAG</td><td> 1-10-1 MOE</td><td> 849</td>

<td> 147037</td><td> 52293</td><td> 52304</td><td> GCCCAGTTCCCA</td><td> 1-10-1 MOE</td><td> 863</td>

<td> 147036</td><td> 52438</td><td> 52449</td><td> CCCAGTTCCCAG</td><td> 1-10-1 MOE</td><td> 849</td>

<td> 147037</td><td> 52439</td><td> 52450</td><td> GCCCAGTTCCCA</td><td> 1-10-1 MOE</td><td> 863</td>

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<td> 147034</td><td> 53148</td><td> 53159</td><td> CAGTTCCCAGCC</td><td> 1-10-1 MOE</td><td> 844</td>

<td> 147034</td><td> 53294</td><td> 53305</td><td> CAGTTCCCAGCC</td><td> 1-10-1 MOE</td><td> 844</td>

<td> 147042</td><td> 53445</td><td> 53456</td><td> GGTCAAAAGGGC</td><td> 1-10-1 MOE</td><td> 866</td>

<td> 147043</td><td> 53446</td><td> 53457</td><td> TGGTCAAAAGGG</td><td> 1-10-1 MOE</td><td> 881</td>

<td> 147044</td><td> 53447</td><td> 53458</td><td> GTGGTCAAAAGG</td><td> 1-10-1 MOE</td><td> 869</td>

<td> 147042</td><td> 53591</td><td> 53602</td><td> GGTCAAAAGGGC</td><td> 1-10-1 MOE</td><td> 866</td>

<td> 147030</td><td> 53592</td><td> 53603</td><td> TCCCAGCCTTGT</td><td> 1-10-1 MOE</td><td> 874</td>

<td> 147043</td><td> 53592</td><td> 53603</td><td> TGGTCAAAAGGG</td><td> 1-10-1 MOE</td><td> 881</td>

<td> 147031</td><td> 53593</td><td> 53604</td><td> TTCCCAGCCTTG</td><td> 1-10-1 MOE</td><td> 885</td>

<td> 147044</td><td> 53593</td><td> 53604</td><td> GTGGTCAAAAGG</td><td> 1-10-1 MOE</td><td> 869</td>

<td> 147030</td><td> 53738</td><td> 53749</td><td> TCCCAGCCTTGT</td><td> 1-10-1 MOE</td><td> 874</td>

<td> 147031</td><td> 53739</td><td> 53750</td><td> TTCCCAGCCTTG</td><td> 1-10-1 MOE</td><td> 885</td>

<td> 147040</td><td> 53783</td><td> 53794</td><td> GCCGCCCAGTTC</td><td> 1-10-1 MOE</td><td> 864</td>

<td> 147041</td><td> 53784</td><td> 53795</td><td> AGCCGCCCAGTT</td><td> 1-10-1 MOE</td><td> 834</td>

<td> 147041</td><td> 53930</td><td> 53941</td><td> AGCCGCCCAGTT</td><td> 1-10-1 MOE</td><td> 834</td>

<td> 147042</td><td> 55008</td><td> 55019</td><td> GGTCAAAAGGGC</td><td> 1-10-1 MOE</td><td> 866</td>

<td> 147043</td><td> 55009</td><td> 55020</td><td> TGGTCAAAAGGG</td><td> 1-10-1 MOE</td><td> 881</td>

<td> 147042</td><td> 55154</td><td> 55165</td><td> GGTCAAAAGGGC</td><td> 1-10-1 MOE</td><td> 866</td>

<td> 147043</td><td> 55155</td><td> 55166</td><td> TGGTCAAAAGGG</td><td> 1-10-1 MOE</td><td> 881</td>

<td> 147058</td><td> 55281</td><td> 55292</td><td> AATTTAATCCGA</td><td> 1-10-1 MOE</td><td> 830</td>

<td> 147058</td><td> 55427</td><td> 55438</td><td> AATTTAATCCGA</td><td> 1-10-1 MOE</td><td> 830</td>

<td> 147019</td><td> 55682</td><td> 55693</td><td> TCGATCTCCTCG</td><td> 1-10-1 MOE</td><td> 877</td>

<td> 147021</td><td> 55684</td><td> 55695</td><td> TGTCGATCTCCT</td><td> 1-10-1 MOE</td><td> 882</td>

<td> 147021</td><td> 55830</td><td> 55841</td><td> TGTCGATCTCCT</td><td> 1-10-1 MOE</td><td> 882</td>

<td> 147054</td><td> 56275</td><td> 56286</td><td> TAATCCGACTGT</td><td> 1-10-1 MOE</td><td> 871</td>

<td> 147055</td><td> 56276</td><td> 56287</td><td> TTAATCCGACTG</td><td> 1-10-1 MOE</td><td> 884</td>

<td> 147056</td><td> 56277</td><td> 56288</td><td> TTTAATCCGACT</td><td> 1-10-1 MOE</td><td> 887</td>

<td> 147058</td><td> 56279</td><td> 56290</td><td> AATTTAATCCGA</td><td> 1-10-1 MOE</td><td> 830</td>

<td> 147059</td><td> 56280</td><td> 56291</td><td> CAATTTAATCCG</td><td> 1-10-1 MOE</td><td> 840</td>

<td> 147060</td><td> 56281</td><td> 56292</td><td> GCAATTTAATCC</td><td> 1-10-1 MOE</td><td> 861</td>

<td> 147061</td><td> 56282</td><td> 56293</td><td> TGCAATTTAATC</td><td> 1-10-1 MOE</td><td> 879</td>

<td> 147051</td><td> 56418</td><td> 56429</td><td> TCCGACTGTGGT</td><td> 1-10-1 MOE</td><td> 875</td>

<td> 147053</td><td> 56420</td><td> 56431</td><td> AATCCGACTGTG</td><td> 1-10-1 MOE</td><td> 829</td>

<td> 147054</td><td> 56421</td><td> 56432</td><td> TAATCCGACTGT</td><td> 1-10-1 MOE</td><td> 871</td>

<td> 147055</td><td> 56422</td><td> 56433</td><td> TTAATCCGACTG</td><td> 1-10-1 MOE</td><td> 884</td>

<td> 147056</td><td> 56423</td><td> 56434</td><td> TTTAATCCGACT</td><td> 1-10-1 MOE</td><td> 887</td>

<td> 147057</td><td> 56424</td><td> 56435</td><td> ATTTAATCCGAC</td><td> 1-10-1 MOE</td><td> 839</td>

<td> 147058</td><td> 56425</td><td> 56436</td><td> AATTTAATCCGA</td><td> 1-10-1 MOE</td><td> 830</td>

<td> 147061</td><td> 56428</td><td> 56439</td><td> TGCAATTTAATC</td><td> 1-10-1 MOE</td><td> 879</td>

<td> 147045</td><td> 57118</td><td> 57129</td><td> TGTGGTCAAAAG</td><td> 1-10-1 MOE</td><td> 883</td>

<td> 147045</td><td> 57264</td><td> 57275</td><td> TGTGGTCAAAAG</td><td> 1-10-1 MOE</td><td> 883</td>

<td> 147046</td><td> 57265</td><td> 57276</td><td> CTGTGGTCAAAA</td><td> 1-10-1 MOE</td><td> 858</td>

<td> 147071</td><td> 58028</td><td> 58039</td><td> CTGATCCTGCAC</td><td> 1-10-1 MOE</td><td> 856</td>

<td> 147071</td><td> 58174</td><td> 58185</td><td> CTGATCCTGCAC</td><td> 1-10-1 MOE</td><td> 856</td>

<td> 147043</td><td> 61111</td><td> 61122</td><td> TGGTCAAAAGGG</td><td> 1-10-1 MOE</td><td> 881</td>

<td> 147071</td><td> 61130</td><td> 61141</td><td> CTGATCCTGCAC</td><td> 1-10-1 MOE</td><td> 856</td>

<td> 147020</td><td> 61226</td><td> 61237</td><td> GTCGATCTCCTC</td><td> 1-10-1 MOE</td><td> 868</td>

<td> 147043</td><td> 61257</td><td> 61268</td><td> TGGTCAAAAGGG</td><td> 1-10-1 MOE</td><td> 881</td>

<td> 147071</td><td> 61276</td><td> 61287</td><td> CTGATCCTGCAC</td><td> 1-10-1 MOE</td><td> 856</td>

<td> 147035</td><td> 61277</td><td> 61288</td><td> CCAGTTCCCAGC</td><td> 1-10-1 MOE</td><td> 847</td>

<td> 147036</td><td> 61278</td><td> 61289</td><td> CCCAGTTCCCAG</td><td> 1-10-1 MOE</td><td> 849</td>

<td> 147037</td><td> 61279</td><td> 61290</td><td> GCCCAGTTCCCA</td><td> 1-10-1 MOE</td><td> 863</td>

<td> 147038</td><td> 61280</td><td> 61291</td><td> CGCCCAGTTCCC</td><td> 1-10-1 MOE</td><td> 855</td>

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<td> 147039</td><td> 61281</td><td> 61292</td><td> CCGCCCAGTTCC</td><td> 1-10-1 MOE</td><td> 850</td>

<td> 147040</td><td> 61282</td><td> 61293</td><td> GCCGCCCAGTTC</td><td> 1-10-1 MOE</td><td> 864</td>

<td> 147071</td><td> 61309</td><td> 61320</td><td> CTGATCCTGCAC</td><td> 1-10-1 MOE</td><td> 856</td>

<td> 147020</td><td> 61372</td><td> 61383</td><td> GTCGATCTCCTC</td><td> 1-10-1 MOE</td><td> 868</td>

<td> 147034</td><td> 61422</td><td> 61433</td><td> CAGTTCCCAGCC</td><td> 1-10-1 MOE</td><td> 844</td>

<td> 147035</td><td> 61423</td><td> 61434</td><td> CCAGTTCCCAGC</td><td> 1-10-1 MOE</td><td> 847</td>

<td> 147036</td><td> 61424</td><td> 61435</td><td> CCCAGTTCCCAG</td><td> 1-10-1 MOE</td><td> 849</td>

<td> 147037</td><td> 61425</td><td> 61436</td><td> GCCCAGTTCCCA</td><td> 1-10-1 MOE</td><td> 863</td>

<td> 147038</td><td> 61426</td><td> 61437</td><td> CGCCCAGTTCCC</td><td> 1-10-1 MOE</td><td> 855</td>

<td> 147040</td><td> 61428</td><td> 61439</td><td> GCCGCCCAGTTC</td><td> 1-10-1 MOE</td><td> 864</td>

<td> 147071</td><td> 61455</td><td> 61466</td><td> CTGATCCTGCAC</td><td> 1-10-1 MOE</td><td> 856</td>

<td> 147073</td><td> 62003</td><td> 62014</td><td> CACTGATCCTGC</td><td> 1-10-1 MOE</td><td> 842</td>

<td> 147073</td><td> 62149</td><td> 62160</td><td> CACTGATCCTGC</td><td> 1-10-1 MOE</td><td> 842</td>

<td> 147066</td><td> 63065</td><td> 63076</td><td> CCTGCACTGACG</td><td> 1-10-1 MOE</td><td> 851</td>

<td> 147068</td><td> 63067</td><td> 63078</td><td> ATCCTGCACTGA</td><td> 1-10-1 MOE</td><td> 838</td>

<td> 147069</td><td> 63146</td><td> 63157</td><td> GATCCTGCACTG</td><td> 1-10-1 MOE</td><td> 860</td>

<td> 147062</td><td> 63207</td><td> 63218</td><td> CACTGACGAGTC</td><td> 1-10-1 MOE</td><td> 841</td>

<td> 147066</td><td> 63211</td><td> 63222</td><td> CCTGCACTGACG</td><td> 1-10-1 MOE</td><td> 851</td>

<td> 147057</td><td> 64054</td><td> 64065</td><td> ATTTAATCCGAC</td><td> 1-10-1 MOE</td><td> 839</td>

<td> 147036</td><td> 64538</td><td> 64549</td><td> CCCAGTTCCCAG</td><td> 1-10-1 MOE</td><td> 849</td>

<td> 147037</td><td> 64539</td><td> 64550</td><td> GCCCAGTTCCCA</td><td> 1-10-1 MOE</td><td> 863</td>

<td> 147037</td><td> 64685</td><td> 64696</td><td> GCCCAGTTCCCA</td><td> 1-10-1 MOE</td><td> 863</td>

<td> 147066</td><td> 64864</td><td> 64875</td><td> CCTGCACTGACG</td><td> 1-10-1 MOE</td><td> 851</td>

<td> 147067</td><td> 64865</td><td> 64876</td><td> TCCTGCACTGAC</td><td> 1-10-1 MOE</td><td> 876</td>

<td> 147066</td><td> 65010</td><td> 65021</td><td> CCTGCACTGACG</td><td> 1-10-1 MOE</td><td> 851</td>

<td> 147067</td><td> 65011</td><td> 65022</td><td> TCCTGCACTGAC</td><td> 1-10-1 MOE</td><td> 876</td>

<td> 147045</td><td> 65017</td><td> 65028</td><td> TGTGGTCAAAAG</td><td> 1-10-1 MOE</td><td> 883</td>

<td> 147045</td><td> 65163</td><td> 65174</td><td> TGTGGTCAAAAG</td><td> 1-10-1 MOE</td><td> 883</td>

<td> 147046</td><td> 65164</td><td> 65175</td><td> CTGTGGTCAAAA</td><td> 1-10-1 MOE</td><td> 858</td>

<td> 147068</td><td> 65408</td><td> 65419</td><td> ATCCTGCACTGA</td><td> 1-10-1 MOE</td><td> 838</td>

<td> 147071</td><td> 65411</td><td> 65422</td><td> CTGATCCTGCAC</td><td> 1-10-1 MOE</td><td> 856</td>

<td> 147069</td><td> 65549</td><td> 65560</td><td> GATCCTGCACTG</td><td> 1-10-1 MOE</td><td> 860</td>

<td> 147068</td><td> 65554</td><td> 65565</td><td> ATCCTGCACTGA</td><td> 1-10-1 MOE</td><td> 838</td>

<td> 147071</td><td> 65557</td><td> 65568</td><td> CTGATCCTGCAC</td><td> 1-10-1 MOE</td><td> 856</td>

<td> 147029</td><td> 67741</td><td> 67752</td><td> CCCAGCCTTGTC</td><td> 1-10-1 MOE</td><td> 848</td>

<td> 147030</td><td> 67742</td><td> 67753</td><td> TCCCAGCCTTGT</td><td> 1-10-1 MOE</td><td> 874</td>

<td> 147031</td><td> 67743</td><td> 67754</td><td> TTCCCAGCCTTG</td><td> 1-10-1 MOE</td><td> 885</td>

<td> 147028</td><td> 67886</td><td> 67897</td><td> CCAGCCTTGTCG</td><td> 1-10-1 MOE</td><td> 846</td>

<td> 147029</td><td> 67887</td><td> 67898</td><td> CCCAGCCTTGTC</td><td> 1-10-1 MOE</td><td> 848</td>

<td> 147030</td><td> 67888</td><td> 67899</td><td> TCCCAGCCTTGT</td><td> 1-10-1 MOE</td><td> 874</td>

<td> 147031</td><td> 67889</td><td> 67900</td><td> TTCCCAGCCTTG</td><td> 1-10-1 MOE</td><td> 885</td>

<td> 147043</td><td> 68867</td><td> 68878</td><td> TGGTCAAAAGGG</td><td> 1-10-1 MOE</td><td> 881</td>

<td> 147044</td><td> 68868</td><td> 68879</td><td> GTGGTCAAAAGG</td><td> 1-10-1 MOE</td><td> 869</td>

<td> 147045</td><td> 68869</td><td> 68880</td><td> TGTGGTCAAAAG</td><td> 1-10-1 MOE</td><td> 883</td>

<td> 147043</td><td> 69013</td><td> 69024</td><td> TGGTCAAAAGGG</td><td> 1-10-1 MOE</td><td> 881</td>

<td> 147044</td><td> 69014</td><td> 69025</td><td> GTGGTCAAAAGG</td><td> 1-10-1 MOE</td><td> 869</td>

<td> 147045</td><td> 69015</td><td> 69026</td><td> TGTGGTCAAAAG</td><td> 1-10-1 MOE</td><td> 883</td>

<td> 147046</td><td> 69016</td><td> 69027</td><td> CTGTGGTCAAAA</td><td> 1-10-1 MOE</td><td> 858</td>

<td> 147071</td><td> 69519</td><td> 69530</td><td> CTGATCCTGCAC</td><td> 1-10-1 MOE</td><td> 856</td>

<td> 147072</td><td> 69520</td><td> 69531</td><td> ACTGATCCTGCA</td><td> 1-10-1 MOE</td><td> 832</td>

<td> 147073</td><td> 69521</td><td> 69532</td><td> CACTGATCCTGC</td><td> 1-10-1 MOE</td><td> 842</td>

<td> 147071</td><td> 69665</td><td> 69676</td><td> CTGATCCTGCAC</td><td> 1-10-1 MOE</td><td> 856</td>

<td> 147072</td><td> 69666</td><td> 69677</td><td> ACTGATCCTGCA</td><td> 1-10-1 MOE</td><td> 832</td>

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<td> 147073</td><td> 69667</td><td> 69678</td><td> CACTGATCCTGC</td><td> 1-10-1 MOE</td><td> 842</td>

<td> 147074</td><td> 69668</td><td> 69679</td><td> CCACTGATCCTG</td><td> 1-10-1 MOE</td><td> 845</td>

<td> 147066</td><td> 69869</td><td> 69880</td><td> CCTGCACTGACG</td><td> 1-10-1 MOE</td><td> 851</td>

<td> 147066</td><td> 70015</td><td> 70026</td><td> CCTGCACTGACG</td><td> 1-10-1 MOE</td><td> 851</td>

<td> 147023</td><td> 70465</td><td> 70476</td><td> CTTGTCGATCTC</td><td> 1-10-1 MOE</td><td> 859</td>

<td> 147023</td><td> 70611</td><td> 70622</td><td> CTTGTCGATCTC</td><td> 1-10-1 MOE</td><td> 859</td>

<td> 147062</td><td> 70615</td><td> 70626</td><td> CACTGACGAGTC</td><td> 1-10-1 MOE</td><td> 841</td>

<td> 147063</td><td> 70616</td><td> 70627</td><td> GCACTGACGAGT</td><td> 1-10-1 MOE</td><td> 862</td>

<td> 147064</td><td> 70617</td><td> 70628</td><td> TGCACTGACGAO</td><td> 1-10-1 MOE</td><td> 880</td>

<td> 147065</td><td> 70618</td><td> 70629</td><td> CTGCACTGACGA</td><td> 1-10-1 MOE</td><td> 857</td>

<td> 147066</td><td> 70619</td><td> 70630</td><td> CCTGCACTGACG</td><td> 1-10-1 MOE</td><td> 851</td>

<td> 147063</td><td> 70762</td><td> 70773</td><td> GCACTGACGAGT</td><td> 1-10-1 MOE</td><td> 862</td>

<td> 147064</td><td> 70763</td><td> 70774</td><td> TGCACTGACGAG</td><td> 1-10-1 MOE</td><td> 880</td>

<td> 147065</td><td> 70764</td><td> 70775</td><td> CTGCACTGACGA</td><td> 1-10-1 MOE</td><td> 857</td>

<td> 147066</td><td> 70765</td><td> 70776</td><td> CCTGCACTGACG</td><td> 1-10-1 MOE</td><td> 851</td>

<td> 147072</td><td> 70998</td><td> 71009</td><td> ACTGATCCTGCA</td><td> 1-10-1 MOE</td><td> 832</td>

<td> 147073</td><td> 70999</td><td> 71010</td><td> CACTGATCCTGC</td><td> 1-10-1 MOE</td><td> 842</td>

<td> 147072</td><td> 71144</td><td> 71155</td><td> ACTGATCCTGCA</td><td> 1-10-1 MOE</td><td> 832</td>

<td> 147073</td><td> 71145</td><td> 71156</td><td> CACTGATCCTGC</td><td> 1-10-1 MOE</td><td> 842</td>

<td> 147074</td><td> 71146</td><td> 71157</td><td> CCACTGATCCTG</td><td> 1-10-1 MOE</td><td> 845</td>

<td> 147037</td><td> 71351</td><td> 71362</td><td> GCCCAGTTCCCA</td><td> 1-10-1 MOE</td><td> 863</td>

<td> 147038</td><td> 71352</td><td> 71363</td><td> CGCCCAGTTCCC</td><td> 1-10-1 MOE</td><td> 855</td>

<td> 147039</td><td> 71353</td><td> 71364</td><td> CCGCCCAGTTCC</td><td> 1-10-1 MOE</td><td> 850</td>

<td> 147037</td><td> 71497</td><td> 71508</td><td> GCCCAGTTCCCA</td><td> 1-10-1 MOE</td><td> 863</td>

<td> 147038</td><td> 71498</td><td> 71509</td><td> CGCCCAGTTCCC</td><td> 1-10-1 MOE</td><td> 855</td>

<td> 147039</td><td> 71499</td><td> 71510</td><td> CCGCCCAGTTCC</td><td> 1-10-1 MOE</td><td> 850</td>

<td> 147061</td><td> 71641</td><td> 71652</td><td> TGCAATTTAATC</td><td> 1-10-1 MOE</td><td> 879</td>

<td> 147061</td><td> 71787</td><td> 71798</td><td> TGCAATTTAATC</td><td> 1-10-1 MOE</td><td> 879</td>

Table 17: Short antisense compounds targeted to SEQ ID NO: 11 and having 1 or 2 mismatches

<td> ISIS NO.</td><td> 5’ Target Site</td><td> 3’ Target Site</td><td> Sequence (5’-3’)</td><td> Gapmer Motif</td><td> SEQ ID NO</td>

<td> 147022</td><td> 177</td><td> 188</td><td> TTGTCGATCTCC</td><td> 1-10-1 MOE</td><td> 886</td>

<td> 147023</td><td> 178</td><td> 189</td><td> CTTGTCGATCTC</td><td> 1-10-1 MOE</td><td> 859</td>

<td> 147020</td><td> 196</td><td> 207</td><td> GTCGATCTCCTC</td><td> 1-10-1 MOE</td><td> 868</td>

<td> 147022</td><td> 198</td><td> 209</td><td> TTGTCGATCTCC</td><td> 1-10-1 MOE</td><td> 886</td>

<td> 147024</td><td> 200</td><td> 211</td><td> CCTTGTCGATCT</td><td> 1-10-1 MOE</td><td> 853</td>

<td> 147026</td><td> 202</td><td> 213</td><td> AGCCTTGTCGAT</td><td> 1-10-1 MOE</td><td> 835</td>

<td> 147028</td><td> 204</td><td> 215</td><td> CCAGCCTTGTCG</td><td> 1-10-1 MOE</td><td> 846</td>

<td> 147029</td><td> 205</td><td> 216</td><td> CCCAGCCTTGTC</td><td> 1-10-1 MOE</td><td> 848</td>

<td> 147030</td><td> 206</td><td> 217</td><td> TCCCAGCCTTGT</td><td> 1-10-1 MOE</td><td> 874</td>

<td> 147036</td><td> 212</td><td> 223</td><td> CCCAGTTCCCAG</td><td> 1-10-1 MOE</td><td> 849</td>

<td> 147073</td><td> 311</td><td> 322</td><td> CACTGATCCTGC</td><td> 1-10-1 MOE</td><td> 842</td>

<td> 147046</td><td> 327</td><td> 338</td><td> CTGTGGTCAAAA</td><td> 1-10-1 MOE</td><td> 858</td>

<td> 147047</td><td> 328</td><td> 339</td><td> ACTGTGGTCAAA</td><td> 1-10-1 MOE</td><td> 833</td>

<td> 147048</td><td> 329</td><td> 340</td><td> GACTGTGGTCAA</td><td> 1-10-1 MOE</td><td> 888</td>

<td> 147049</td><td> 330</td><td> 341</td><td> CGACTGTGGTCA</td><td> 1-10-1 MOE</td><td> 854</td>

<td> 147050</td><td> 331</td><td> 342</td><td> CCGACTGTGGTC</td><td> 1-10-1 MOE</td><td> 889</td>

<td> 147051</td><td> 332</td><td> 343</td><td> TCCGACTGTGGT</td><td> 1-10-1 MOE</td><td> 875</td>

<td> 147052</td><td> 333</td><td> 344</td><td> ATCCGACTGTGG</td><td> 1-10-1 MOE</td><td> 837</td>

<td> 147053</td><td> 334</td><td> 345</td><td> AATCCGACTGTG</td><td> 1-10-1 MOE</td><td> 829</td>

<td> 147054</td><td> 335</td><td> 346</td><td> TAATCCGACTGT</td><td> 1-10-1 MOE</td><td> 871</td>

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<td> 147055</td><td> 336</td><td> 347</td><td> TTAATCCGACTG</td><td> 1-10-1 MOE</td><td> 884</td>

<td> 147056</td><td> 337</td><td> 348</td><td> TTTAATCCGACT</td><td> 1-10-1 MOE</td><td> 887</td>

<td> 147057</td><td> 338</td><td> 349</td><td> ATTTAATCCGAC</td><td> 1-10-1 MOE</td><td> 839</td>

<td> 147058</td><td> 339</td><td> 350</td><td> AATTTAATCCGA</td><td> 1-10-1 MOE</td><td> 830</td>

<td> 147060</td><td> 341</td><td> 352</td><td> GCAATTTAATCC</td><td> 1-10-1 MOE</td><td> 861</td>

<td> 147061</td><td> 342</td><td> 353</td><td> TGCAATTTAATC</td><td> 1-10-1 MOE</td><td> 879</td>

<td> 147062</td><td> 1024</td><td> 1035</td><td> CACTGACGAGTC</td><td> 1-10-1 MOE</td><td> 841</td>

<td> 147063</td><td> 1025</td><td> 1036</td><td> GCACTGACGAGT</td><td> 1-10-1 MOE</td><td> 862</td>

<td> 147068</td><td> 1030</td><td> 1041</td><td> ATCCTGCACTGA</td><td> 1-10-1 MOE</td><td> 838</td>

<td> 147071</td><td> 1033</td><td> 1044</td><td> CTGATCCTGCAC</td><td> 1-10-1 MOE</td><td> 856</td>

<td> 147073</td><td> 1035</td><td> 1046</td><td> CACTGATCCTGC</td><td> 1-10-1 MOE</td><td> 842</td>

<td> 147074</td><td> 1036</td><td> 1047</td><td> CCACTGATCCTG</td><td> 1-10-1 MOE</td><td> 845</td>

<td> 147067</td><td> 1091</td><td> 1102</td><td> TCCTGCACTGAC</td><td> 1-10-1 MOE</td><td> 876</td>

<td> 147024</td><td> 1891</td><td> 1902</td><td> CCTTGTCGATCT</td><td> 1-10-1 MOE</td><td> 853</td>

<td> 147026</td><td> 1893</td><td> 1904</td><td> AGCCTTGTCGAT</td><td> 1-10-1 MOE</td><td> 835</td>

<td> 147029</td><td> 1896</td><td> 1907</td><td> CCCAGCCTTGTC</td><td> 1-10-1 MOE</td><td> 848</td>

<td> 147036</td><td> 1903</td><td> 1914</td><td> CCCAGTTCCCAG</td><td> 1-10-1 MOE</td><td> 849</td>

<td> 147039</td><td> 1906</td><td> 1917</td><td> CCGCCCAGTTCC</td><td> 1-10-1 MOE</td><td> 850</td>

<td> 147019</td><td> 1994</td><td> 2005</td><td> TCGATCTCCTCG</td><td> 1-10-1 MOE</td><td> 877</td>

<td> 401385</td><td> 2815</td><td> 2828</td><td> CCCAGTGGGTTTGA</td><td> 2-10-2 MOE</td><td> 890</td>

<td> 147033</td><td> 5265</td><td> 5276</td><td> AGTTCCCAGCCT</td><td> 1-10-1 MOE</td><td> 836</td>

<td> 147033</td><td> 5373</td><td> 5384</td><td> AGTTCCCAGCCT</td><td> 1-10-1 MOE</td><td> 836</td>

<td> 147060</td><td> 7168</td><td> 7179</td><td> GCAATTTAATCC</td><td> 1-10-1 MOE</td><td> 861</td>

<td> 147053</td><td> 10527</td><td> 10538</td><td> AATCCGACTGTG</td><td> 1-10-1 MOE</td><td> 829</td>

<td> 147053</td><td> 10635</td><td> 10646</td><td> AATCCGACTGTG</td><td> 1-10-1 MOE</td><td> 829</td>

<td> 147070</td><td> 11604</td><td> 11615</td><td> TGATCCTGCACT</td><td> 1-10-1 MOE</td><td> 878</td>

<td> 147071</td><td> 11612</td><td> 11623</td><td> CTGATCCTGCAC</td><td> 1-10-1 MOE</td><td> 856</td>

<td> 147072</td><td> 12294</td><td> 12305</td><td> ACTGATCCTGCA</td><td> 1-10-1 MOE</td><td> 832</td>

<td> 147072</td><td> 12299</td><td> 12310</td><td> ACTGATCCTGCA</td><td> 1-10-1 MOE</td><td> 832</td>

<td> 147052</td><td> 12938</td><td> 12949</td><td> ATCCGACTGTGG</td><td> 1-10-1 MOE</td><td> 837</td>

<td> 147052</td><td> 13119</td><td> 13130</td><td> ATCCGACTGTGG</td><td> 1-10-1 MOE</td><td> 837</td>

<td> 147047</td><td> 13158</td><td> 13169</td><td> ACTGTGGTCAAA</td><td> 1-10-1 MOE</td><td> 833</td>

<td> 147048</td><td> 13159</td><td> 13170</td><td> GACTGTGGTCAA</td><td> 1-10-1 MOE</td><td> 888</td>

<td> 147049</td><td> 13160</td><td> 13171</td><td> CGACTGTGGTCA</td><td> 1-10-1 MOE</td><td> 854</td>

<td> 147048</td><td> 13340</td><td> 13351</td><td> GACTGTGGTCAA</td><td> 1-10-1 MOE</td><td> 888</td>

<td> 147049</td><td> 13341</td><td> 13352</td><td> CGACTGTGGTCA</td><td> 1-10-1 MOE</td><td> 854</td>

<td> 147051</td><td> 13343</td><td> 13354</td><td> TCCGACTGTGGT</td><td> 1-10-1 MOE</td><td> 875</td>

<td> 147061</td><td> 13497</td><td> 13508</td><td> TGCAATTTAATC</td><td> 1-10-1 MOE</td><td> 879</td>

<td> 147069</td><td> 15562</td><td> 15573</td><td> GATCCTGCACTG</td><td> 1-10-1 MOE</td><td> 860</td>

<td> 147068</td><td> 15743</td><td> 15754</td><td> ATCCTGCACTGA</td><td> 1-10-1 MOE</td><td> 838</td>

<td> 147049</td><td> 17181</td><td> 17192</td><td> CGACTGTGGTCA</td><td> 1-10-1 MOE</td><td> 854</td>

<td> 147049</td><td> 17349</td><td> 17360</td><td> CGACTGTGGTCA</td><td> 1-10-1 MOE</td><td> 854</td>

<td> 147047</td><td> 22438</td><td> 22449</td><td> ACTGTGGTCAAA</td><td> 1-10-1 MOE</td><td> 833</td>

<td> 147047</td><td> 24322</td><td> 24333</td><td> ACTGTGGTCAAA</td><td> 1-10-1 MOE</td><td> 833</td>

<td> 147045</td><td> 24488</td><td> 24499</td><td> TGTGGTCAAAAG</td><td> 1-10-1 MOE</td><td> 883</td>

<td> 147039</td><td> 25064</td><td> 25075</td><td> CCGCCCAGTTCC</td><td> 1-10-1 MOE</td><td> 850</td>

<td> 147057</td><td> 25508</td><td> 25519</td><td> ATTTAATCCGAC</td><td> 1-10-1 MOE</td><td> 839</td>

<td> 147057</td><td> 25676</td><td> 25687</td><td> ATTTAATCCGAC</td><td> 1-10-1 MOE</td><td> 839</td>

<td> 147061</td><td> 25680</td><td> 25691</td><td> TGCAATTTAATC</td><td> 1-10-1 MOE</td><td> 879</td>

<td> 147069</td><td> 28731</td><td> 28742</td><td> GATCCTGCACTG</td><td> 1-10-1 MOE</td><td> 860</td>

<td> 147052</td><td> 30132</td><td> 30143</td><td> ATCCGACTGTGG</td><td> 1-10-1 MOE</td><td> 837</td>

<td> 147052</td><td> 30277</td><td> 30288</td><td> ATCCGACTGTGG</td><td> 1-10-1 MOE</td><td> 837</td>

<td> 147036</td><td> 32085</td><td> 32096</td><td> CCCAGTTCCCAG</td><td> 1-10-1 MOE</td><td> 849</td>

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<td> 147072</td><td> 32520</td><td> 32531</td><td> ACTGATCCTGCA</td><td> 1-10-1 MOE</td><td> 832</td>

<td> 147071</td><td> 33058</td><td> 33069</td><td> CTGATCCTGCAC</td><td> 1-10-1 MOE</td><td> 856</td>

<td> 147050</td><td> 33125</td><td> 33136</td><td> CCGACTGTGGTC</td><td> 1-10-1 MOE</td><td> 889</td>

<td> 147069</td><td> 33204</td><td> 33215</td><td> GATCCTGCACTG</td><td> 1-10-1 MOE</td><td> 860</td>

<td> 147050</td><td> 33273</td><td> 33284</td><td> CCGACTGTGGTC</td><td> 1-10-1 MOE</td><td> 889</td>

<td> 147047</td><td> 33319</td><td> 33330</td><td> ACTGTGGTCAAA</td><td> 1-10-1 MOE</td><td> 833</td>

<td> 147050</td><td> 33322</td><td> 33333</td><td> CCGACTGTGGTC</td><td> 1-10-1 MOE</td><td> 889</td>

<td> 147052</td><td> 33324</td><td> 33335</td><td> ATCCGACTGTGG</td><td> 1-10-1 MOE</td><td> 837</td>

<td> 147049</td><td> 33469</td><td> 33480</td><td> CGACTGTGGTCA</td><td> 1-10-1 MOE</td><td> 854</td>

<td> 147050</td><td> 33470</td><td> 33481</td><td> CCGACTGTGGTC</td><td> 1-10-1 MOE</td><td> 889</td>

<td> 147052</td><td> 33472</td><td> 33483</td><td> ATCCGACTGTGG</td><td> 1-10-1 MOE</td><td> 837</td>

<td> 147047</td><td> 33641</td><td> 33652</td><td> ACTGTGGTCAAA</td><td> 1-10-1 MOE</td><td> 833</td>

<td> 147047</td><td> 33789</td><td> 33800</td><td> ACTGTGGTCAAA</td><td> 1-10-1 MOE</td><td> 833</td>

<td> 147059</td><td> 35585</td><td> 35596</td><td> CAATTTAATCCG</td><td> 1-10-1 MOE</td><td> 840</td>

<td> 147021</td><td> 36241</td><td> 36252</td><td> TGTCGATCTCCT</td><td> 1-10-1 MOE</td><td> 882</td>

<td> 147073</td><td> 37633</td><td> 37644</td><td> CACTGATCCTGC</td><td> 1-10-1 MOE</td><td> 842</td>

<td> 147033</td><td> 42529</td><td> 42540</td><td> AGTTCCCAGCCT</td><td> 1-10-1 MOE</td><td> 836</td>

<td> 147050</td><td> 45401</td><td> 45412</td><td> CCGACTGTGGTC</td><td> 1-10-1 MOE</td><td> 889</td>

<td> 147050</td><td> 45549</td><td> 45560</td><td> CCGACTGTGGTC</td><td> 1-10-1 MOE</td><td> 889</td>

<td> 147074</td><td> 46125</td><td> 46136</td><td> CCACTGATCCTG</td><td> 1-10-1 MOE</td><td> 845</td>

<td> 147057</td><td> 46313</td><td> 46324</td><td> ATTTAATCCGAC</td><td> 1-10-1 MOE</td><td> 839</td>

<td> 147058</td><td> 46462</td><td> 46473</td><td> AATTTAATCCGA</td><td> 1-10-1 MOE</td><td> 830</td>

<td> 147058</td><td> 47413</td><td> 47424</td><td> AATTTAATCCGA</td><td> 1-10-1 MOE</td><td> 830</td>

<td> 147058</td><td> 47561</td><td> 47572</td><td> AATTTAATCCGA</td><td> 1-10-1 MOE</td><td> 830</td>

<td> 147073</td><td> 48221</td><td> 48232</td><td> CACTGATCCTGC</td><td> 1-10-1 MOE</td><td> 842</td>

<td> 147073</td><td> 48369</td><td> 48380</td><td> CACTGATCCTGC</td><td> 1-10-1 MOE</td><td> 842</td>

<td> 147028</td><td> 48567</td><td> 48578</td><td> CCAGCCTTGTCG</td><td> 1-10-1 MOE</td><td> 846</td>

<td> 147068</td><td> 49199</td><td> 49210</td><td> ATCCTGCACTGA</td><td> 1-10-1 MOE</td><td> 838</td>