Pharmaceutical compositions and related methods of delivery

PHARMACEUTICAL COMPOSITIONS AND RELATED METHODS OF DELIVERY

FIELD OF THE TECHNOLOGY

The present invention relates generally to pharmaceutical compositions enabling improved delivery e.g. oral delivery and methods of using such compositions.

BACKGROUND

Techniques enabling efficient transfer of a substance of interest across a biological barrier are of considerable interest in the fields of biotechnology and medicine. For example, such techniques may be used for the transport of a variety of different substances across a biological barrier regulated by tight junctions (i.e., the mucosal epithelia, which include the intestinal and respiratory epithelia, and the vascular endothelia, which include the blood-brain barrier, nasal membrane, cornea and other eye membranes, and genito-urinary membranes). In particular there is great interest in oral delivery of therapeutic agents to avoid the use of more invasive means of administration and hence improve patient convenience and compliance.

Diverse drug delivery vehicles have been employed, among them liposomes, lipidic or polymeric nanoparticles, and microemulsions. These have improved the oral bioavailability of certain drugs, mostly by the protective effect they offer. However, for most relevant drugs, bioavailability remains very low and fails to achieve the minimal therapeutic goals.

Hence, a need exists for an efficient, specific, non-invasive, low-risk means to target various biological barriers for the non invasive delivery of various therapeutic agents such as peptides and polypeptides, macromolecule drugs and other therapeutic agents which include small molecules with low bioavailability. 2

SUMMARY

This invention provides a pharmaceutical composition comprising a suspension which comprises an admixture of a hydrophobic medium and a solid form wherein the solid form comprises a therapeutically effective amount of octreotide, at least one salt of a medium chain fatty acid and a matrix forming polymer, and wherein the medium chain fatty acid salt is present in the composition at an amount of at least 11% by weight.

In some embodiments, the solid form comprises a particle and/or powder.

In some embodiments, the particle and/or powder is produced by lyophilization or by granulation.

In some embodiments, the solid form comprises a binder.

In some embodiments, the water content p is lower than 6 % by weight.

In some embodiments, the water content composition is lower than 2 % by weight.

In some embodiments, the water content in the solid form is lower than 16% by weight.

In some embodiments, the water content in the solid form is lower than 2% by weight.

In some embodiments, the medium chain fatty acid salt has a chain length from 6 to 14 carbon atoms.

In some embodiments, the medium chain fatty acid salt is sodium hexanoate, sodium heptanoate, sodium octanoate, sodium nonanoate, sodium decanoate, sodium undecanoate, sodium dodecanoate, sodium tridecanoate or sodium tetradecanoate, or a corresponding potassium or lithium or ammonium salt or a combination thereof.

In some embodiments, the fatty acid salt is sodium octanoate.

In some embodiments, the medium chain fatty acid salt is present in the composition at an amount of 11% to 40% by weight.

In some embodiments, the medium chain fatty acid salt is present in the composition at an amount of 12% to 18% by weight

In some embodiments, the medium chain of fatty acid salt is 15% by weight.

In some embodiments, the medium chain fatty acid salt is present in the solid form at an amount of 50% to 90% by weight.

In some embodiments, the medium chain aftty acid salt is present in the solid form at an amount of 70% to 80% by weight.

In some embodiments, the matrix forming polymer is dextran or polyvinylpyrrolidone (PVP).

In some embodiments, the polyvinylpyrrolidone is present in the composition at an amount of 2% to 20% by weight.

In some embodiments, the polyvinylpyrrolidone is present in the composition at an amount of 5% to 15 %by weight.

In some embodiments, the polyvinylpyrrolidone is present in the composition at an amount of 10 % by weight.

In some embodiments, the polyvinylpyrrolidone is PVP-12.

In some embodiments, the polyvinylpyrrolidone has a molecular weight of about 3000.

In some embodiments, the composition is free of a medium chain alcohol.

In some embodiments, the composition is free of a membrane fluidizing agent.

In some embodiments, the hydrophobic medium comprises castor oil or glyceryl tricaprylate or glyceryl tributyrate or a combination thereof.

In some embodiments, the pharmaceutical composition additionally comprises octanoic acid.

In some embodiments, the main component by weight of the hydrophobic medium is castor oil.

In some embodiments, the hydrophobic medium comprises additionally glyceryl tricaprylate.

In some embodiments, the main component by weight of the hydrophobic medium is glyceryl tricaprylate.

In some embodiments, the hydrophobic medium comprises additionally castor oil.

In some embodiments, the hydrophobic medium comprises an aliphatic, olefinic, cyclic or aromatic compound.

In some embodiments, the hydrophobic medium comprises an aliphatic compound.

In some embodiments, the hydrophobic medium comprises a mineral oil, a paraffin, a fatty acid such as octanoic acid, a monoglyceride, a diglyceride, a triglyceride, an ether or an ester, or a combination thereof.

In some embodiments, the triglyceride is a long chain triglyceride, a medium chain triglyceride or a short chain triglyceride.

In some embodiments, the triglyceride is a long chain triglyceride.

In some embodiments, the long chain triglyceride is castor oil or coconut oil or a combination thereof.

In some embodiments, the ester in the hydrophobic medium is a low molecular weight ester.

In some embodiments, the low molecular weight ester is ethyl isovalerate or butyl acetate.

In some embodiments, the triglyceride is a short chain triglyceride or a medium chain triglyceride or a mixture thereof.

In some embodiments, the short chain triglyceride is glyceryl tributyrate and the medium chain triglyceride is glyceryl tricaprylate.

In some embodiments, the hydrophobic medium further comprises an ionic surfactant or a non-ionic surfactant.

In some embodiments, the surfactant is lecithin or a bile salt or a detergent.

In some embodiments, the surfactant is a monoglyceride , a cremophore, a polyethylene glycol fatty alcohol ether, a sorbitan fatty acid ester, a polyoxyethylene sorbitan fatty acid ester, polyoxy ethylene esters of 12-hydroxystearic acid), or a poloxamer or a combination thereof.

In some embodiments, themonoglyceride is glyceryl monocaprylate, glyceryl monoocatnoate, glyceryl monodecanoate, glyceryl monolaurate, glyceryl monomyristate, glyceryl monopalmitate or glyceryl monooleate or glyceryl monostearate or a combination thereof.

In some embodiments, the sorbitan fatty acid ester comprises sorbitan monolaurate, sorbitan monooleate or sorbitan monopalmitate or a combination thereof.

In some embodiments the polyoxyethylene sorbitan fatty acid ester comprises polyoxyethylene sorbitan monooleate, polyoxyethylene sorbitan monostearate or polyoxyethylene sorbitan monopalmitate or a combination thereof.

In some embodiments the composition consists essentially of a therapeutic agent and a medium chain fatty acid salt and a hydrophobic medium.

In some embodiments, the solid form consists essentially of octreotide, a matrix forming polymer and a medium chain fatty acid salt.

In some embodiments, the hydrophobic medium consists essentially of glyceryl tricaprylate.

This invention provides a pharmaceutical composition comprising a suspension which comprises an admixture of a hydrophobic medium and a solid form wherein the solid form comprises a therapeutically effective amount of octreotide, at least one salt of a medium chain fatty acid and a matrix forming polymer, and wherein the matrix forming polymer is present in the composition at an amount of 3% or more by weight.

In some embodiments, the the solid form comprises a particle and/or powder.

In some embodiments the the matrix forming polymer is dextran or polyvinylpyrrolidone (PVP).

In some embodiments, medium chain fatty acid salt is sodium hexanoate, sodium heptanoate, sodium octanoate, sodium nonanoate, sodium decanoate, sodium undecanoate, sodium dodecanoate, sodium tridecanoate or sodium tetradecanoate, or corresponding potassium or lithium or ammonium salt or a combination thereof.

In some embodiments, the the fatty acid salt is sodium octanoate.

In some embodiments, the medium chain fatty acid salt is present in the composition at an amount of 11% to 40% by weight.

In some embodiments, the medium chain fatty acid salt is present in the composition at an amount of 12% to 28% by weight.

In some embodiments, the medium chain fatty acid is 15% by weight.

In some embodiments, the hydrophobic medium comprises a mineral oil, a paraffin, a fatty acid, a monoglyceride, a diglyceride, a triglyceride, an ether or an ester, or a combination thereof.

In some embodiments, the fatty acid is octanoic acid.

BRIEF DESCRIPTION OF THE DRAWINGS

Various aspects of at least one embodiment are discussed below with reference to the accompanying Figures. In the Figures, which are not intended to be drawn to scale, each identical or nearly identical component that is illustrated in various figures is represented by a like numeral. For purposes of clarity, not every component may be labeled in every drawing. The Figures are provided for the purposes of illustration and WO 20111/(13214(1 PCT/IB 2000/007155 7 explanation and are not intended as a definition of the limits of the invention. In the Figures: PIG. 1 presents a process for production of an insulin formulation of a composition in accordance with one or more embodiments as referenced in the 5 accompanying Examples; FIGS. 2-5 present data referenced in accompanying Examples 3 through 6; FIG. 6 presents data referenced in accompanying Example 8; FIG, 7 presents molecular weight marker permeability data tcfercnced in accompanying Example 33: 10 FIG. 8 presents time-course permeability data referenced in accompanying

Example 34; and FIGS. 9 and 10 present data relating to administration οΓ octreotide to monkeys referenced in accompanying Example 35.

15 DETAILED DESCRIPTION

The compositions described herein can be administered to a subject to provide for improved bioavailability of a therapeutic agent

Pharmaceutical compositions: The pharmaceutical compositions described herein include a therapeutic agent and a medium chain fatty acid salt in intimate contact 20 or association with a substantially hydrophobic medium. For example, the therapeutic agent and the medium chain fatty acid or derivative thereof may be coated, suspended, sprayed by or immersed in a substantially hydrophobic medium forming a suspension. The compositions of the invention are not emulsions. Almost all of the compositions are oiiy suspensions and die amount of water in the compositions is very low; a few of the 25 present compositions which are not suspensions incorporate a high amount (about 78% octanoic acid) and are solutions by visual analysis. The suspension may be a liquid suspension incorporating solid material, or a semi-solid suspension incorporating solid material (an ointment).

Many of the compositions described herein comprise a suspension which 30 comprises an admixture of a hydrophobic medium and a solid form wherein the solid form comprises a therapeutically effective amount of a therapeutic agent and at least one salt of a medium chain fatty acid, and wherein the medium chain fatty acid salt is present WO 2« 1 »/«3214« ΚΊ71Β2««<Ι/ΙΚ»7155 8 in the composition at an amount of 10% or more by weight. The solid form may comprise a panicle (e.g. consist essentially of.particles, or consist of panicles). 'Ihe panicle may be produced by lyophilization or by granulation. In some embodiments, preferably after milling. 90% (v/v) of the particles ure below 130 microns, and 50% (v/v) of the panicles 5 are below 45 microns, Λ cargo compound is a therupeutic agent (e.g. insulin) or a test compound fe.g, high molecular weight dextran) which is formulated as described herein within the compositions of the invention.

The inventors were particular to include in many of the compositions of the 10 invention only excipients which are generally recognized as safe, based on available data on human use. animal safety and regulatory guidelines (e.g, GRAS excipients). Some compositions of the invention may have other types of excipients (e.g. non-GRAS). In some embodiments the compositions of the invention huve amounts ol excipients that are within the maximum daily doses us noted in such available data for each specific 15 excipient.

The medium chain fatLy acid salt may generally facilitate or enhance permeability and/or absorption of the therapeutic agent In some embodiments tire medium chain fatty acid salts include derivatives of medium chain fatty acid salts. The therapeutic agent and the medium chain fatty acid salt are in solid farm, for example, a solid particle such as a 20 lyophilized particle, granulated particle, pellet or micro-sphere. In preferred embodiments, the therapeutic agent and the medium chain fatty acid salt are both in the same solid fonn, e.g., both in the same panicle. In other embodiments. the lheraj>eutie agent and the medium chain fatty acid salt may each be in a different solid form. e.g. each in a distinct particle. The compositions descrihed herein are. substantially free of any 25 “membrane fluidizing agents" defined as linear, branched, aromatic and cyclic medium chain alcohols, in particular geraniol and octanol. For example the compositions preferably include no membrane fluidizing agents but certain embodiments may include for example less than 1 % or less than 0.5% or less than 0,1 % by weight of membrane fluidizing agents. 30 Unlike emulsions, where water is an essential constituent of the formulation. the compost lions described herein provide a solid form such as a particle containing the therapeutic agent, which is then associated with the hydrophobic (oily) medium. The WO 2010/03214» PO7IB201I9/IHI7155 9 amount of water in the compositions is generally less than 3% by weight, usually less than about 2% or about 1 % or less by weight, 'Phe compositions described herein are suspensions which comprise an admixture of a hydrophobic medium and a solid form wherein die solid form comprises a 5 therapeuLically effective amount of a therapeutic agent and at least one salt of a medium chain fatty acid. The solid form may be a particle (e.g,, consist essentially of particles, or consist of particles). 'Hie particle may be produced by lyophilization or by granulation.

The medium chain fall)· acid salt is generally present in die compositions described herein at an amount of 10% or more by weight. In certain embodiments the medium chain fatty 10 acid salt is present in the composition at an amount of 10%-50%. preferably 11 %-1 8% or about 11%-17% or 12%-16% or 12%-t5% or 13%-16% or 13%-15%or 14%-16% or I4%-15% or 15%-16% or most preferably 15% or 16% by weight, and the medium chain fatty acid has a chain length from about 6 to about 14 carbon atoms preferably 8. 9 or 10 carbon atoms. 15 In some embodiments in the compositions described above, the solid form including the therapeutic agent also includes a stabilizer (e.g. u stabilizer of protein structure). Stabilizers of protein structure are compounds that stabilize protein structure under aqueous or non-aqueous conditions or can reduce or prevent aggregation of the therapeutic agent, for example during a drying process such as lyophilization or other 20 processing step. Stabilizers of structure can be polyanionic molecules, such as phytic acid, polyvalent ions such as Ca, Z.n or Mg, saccharides such as a di saccharide (e.g., trehalose, maltose) or an oligo or polysaccharide such as dextrin or dextran, or a sugar alcohol such as mannitol, or an amino acid such as glycine, or poivcationic molecules, such as spermine, or surfactants such as polyoxyethylene sorbitan monooleate ( Tween 80) 25 or pluronic acid. Uncharged polymers, such as mannitol, methyl cellulose and polyvinyl alcohol, are also suitable stabilizers.

Although polyvinylpyrrolidone (PVP) is known in the art as a stabilizer, the inventors unexpectedly found that, in die compositions of the invention described herein, PVP. in particular PVP-12. serves to increase die effect of die permeability enhancer in a 30 synergistic manner; furthermore, increasing the level of PVP-12 to I ()% increased the absorption of the therapeutic agent into the blood due to the improved activity of the formulations. The inventors demonstrated that dextran had a similar (but lower) effect as PVP did. Other matrix forming polymers have a similar effect. WO 2010/03214« RCT/1B2009/007155 10 111 some embodiments, such as when the therapeutic agent is a small molecule, a bulking agent may be added, for example, mannitol or glyein.

In certain embodiments of Ute compositions described herein die therapeutic agent is a protein, a polypcpLide, a peptide, a glycosaminoglvean, a small molecule, a 5 polysaccharide ora polynucleotide inter alia, such as octreotide, growth hormone, parathyroid hormone, parathyroid hormone amino acids 1-34 |PTH(l-34) termed teriparalideh a low molecular weight heparin or fondaparinux inter alia. Low molecular weight heparins are defined as heparin salts having an average molecular weight of less than 8(100 Da and for which at least 60% of all chains have a molecular weight less than 10 8000 Da.

In a particular embodiment of the compositions described herein the salt of the fatty acid is sodium octanoate and die hydrophobic medium is castor oil; in another particular embodiment the composition further comprises glyceryl monooieate and sorbitan monopalmitate or glyceryl monocaprylate and glyceryl tricaprylate and 15 polyoxyeihylenesorbitan monooieate; in another particular embodiment the composition further comprises glyceryl tributyrate. lecithin, ethylisovaleraie and at least one stabilizer. In particular embodiments the therapeutic agent is octreotide, growth hormone, parathyroid hormone, teriparatide, interferon-alia (lPN-α). a low molecular weight heparin, fondaparinux, siRNA. somatostatin and analogs (agonises) thereof including 20 pepudomimetics, exenatide. vancomycin or gentamicin inter alia.

Therapeutic agents: 'Ihe pharmaceutical compositions described herein can be used with a variety of therapeutic agents (also termed active pharmaceutical ingredient =API). In some embodiments, the pharmaceutical composition includes a plurality of therapeutic agents 25 (effectors). 'Hie therapeutic agents can either be in the same solid form (e.g., in the same particle), or the therapeutic agents can each be in an independent solid font) (e.g., each in different panicles. In some embodiments, the therapeutic agent is in the form of a panicle, for example, a granulated or solid panicle, The particle is associated with or is in intimate contact with a substantially hydrophobic medium, for example, a hydrophobic 30 medium described herein. 'iherapeutic agents that can be used in the compositions described herein include any molecule or compound senring as, for example, a biological, therapeut ic, pharmaceutical. or diagnostic agent including an imaging agent. The therapeutic agents WO 2010/03214(1 include drugs and other agents including, but not limited to, those listed in the United Stales Pharmacopeia and in other known pharmacopeias. 'iherapeutic agents are incorporated into the formulations of die invention without any chemical modification. Therapeutic agents include proteins, polypeptides, peptides, polynucleotides, 5 polysaccharides and small molecules.

The term "small molecule" is understood to refer to a low molecular weight organic compound which may be synthetically produced or obtained front natural sources and typically has a molecular weight of less than 2000 Da. or less than 1000 Da or even less than 600 Da e.g, less dian or about 550 Da or less than or about 500 Da or less than 10 or about 400 Da: or about 400 Da to about 2000 Da; or about 400 Da io about 1700 Da. Examples of small molecules are ergotamine (molecular weight =582 Da), fondapartnux (molecular weight = 1727 Da), leuprolide (molecular weight = 1209 Da), vancomycin (molecular weight = 1449 Du), gentamicin (molecular weight = 478 Da) and doxorubicin (molecular weight =544). ! 5 The term "polynucleotide" refers io any molecule composed of DNA nucleotides, RNA nucleotides ora combination of both types which comprises two or more of the bases guanidine, cilosine. timidine, adenine, uracil or inostne. inter alia. A polynucleotide may include natural nucleotides, chemically modified nucleotides and synthetic nucleotides, or chemical analogs thereof and may be single-stranded or double- stranded. 20 'Fite tenn includes "oligonucleotides" and encompasses "nucleic acids".

By "small interfering RNA" (siRNA) is meant an RNA molecule (ribonucleotide) which decreases or silences (prevents) the expression of a gene/ niRNA of its endogenous or cellular counterpart. Ilie term is understood to encompass "RNA interference” (RNAi’l, and "double-stranded RNA" (dsRNA). 25 By "polypeptide" is meant a molecule composed of covalently Jinked amino acids and the term includes peptides, polypeptides, proteins and peptidomimetics. A peptidoinimciic is a compound containing non-peptidic structural elements that is capable of mimicking the biological aetion(s) of a natural parent peptide. Some of the classical peptide characteristics such as enzymaLically scissile peptidic bonds are normally not 30 present in a pcpiidoniimeiic.

Ihe term "amino acid" refers to a molecule which consists of any one of the 20 naturally WO 21)10/03214(1 occurring amino acids, amino acids which have been chemically modified or synthetic amino acids.

By “polysaccharide” is mean) a linear or branched polymer composed of covalently linked monosaccharides', glucose is the most common monosaccharide and 5 them are normally at least eight monosaccharide units in a polysaccharide and usually many more. Polysaccharides have a general formula of Cx(H2O)y where x is usually a large number between 200 and 2500, Considering that the repeating units in the polymer backbone are often six-carbon monosaccharides, the general formula can also be represented as (C6H10O5)n where 40<n<3000 i.e. there are normally between 40 and 10 3000 monosaccharide units in a polysaccharide. A “glycosaminoglycan" is a polysaccharide dial contains amino containing sugars.

Exemplary anionic therapeutic agents include polynucleotides from various origins, and particularly from human, viral, animal, eukaryotic or prokaryotic,

15 plant, or synthetic origin, etc including systems for therapeutic gene deliver)'. A polynucleotide of interest may be of a variety of sizes, ranging from, for example, a simple, trace nucleotide to a gene fragment, or an entire gene. It may be a viral gene or a plasmid. Exemplary polynucleotides serving as therapeutic agents include specific DNA sequences (e.g. coding genes), specific RNA sequences (e.g,, RNA aptamers. antisense 20 RNA. short interfering RNA (siRNA) or a specific inhibiiory RNA (RNAi)J. poly CPG. or poly f:C synthetic polymers of polynucleotides.

Alternatively, the therapeutic agenL can be a protein, such as, for example, an enzyme, a hormone, an incretin, a proteoglycan, a ribozyme, a cytokine, a peptide, an ajxilipoprotein, a growth factor, a bioacuve molecule, an antigen, or an antibody or 25 fragmentis) thereof, etc. ’Hie peptide can be a small peptide e.g. from about. 2 to about 4() amino acids, examples include fibrinogen-receptor antagonists (RGD-containing peptides which ate tetrapeptides having an average molecular weight of about 600. Exemplary' peptides are somatostatin and analogs thereof e.g. octreotide and Ianreotitle (Somatulinc) which are both cyclic octapcptides and pasireotide (SOM-230) which is a cyclic 30 hexupeptide (Wcckbecker et al, 2002, Endocrinology 143(10) 4123-4130; Schmid. 2007. Molecular and Cellular Endocrinology 286. 69-74). Other exemplary peptides are glariramer acetate (Copaxone®) which is a tetrapeptide, terltpressin which is a 12 amino acid peptide analog (agonist) of lysine vasopressin (ADH) and exenatide, a 39 amino acid WO 21)1(1/032140 PC'lVlIi2009/007155 13 peptide which is an incretin mimetic agent, and oilier analogs of glucagon-like peptide-l(GIJ’-l). (Byetta® is the trade name forexenatide (Eli Lally and Company / Aiuylin Pharmaceuticals. Inc.). Other peptides include dalargin which is a hexapeptide, and kyotorphin which is a dipeptide. Peptides include growth hormone releasing peptides 5 which are peptides of about 12 amino acids or less: see for example peptides disclosed in US Patent numbers 4411890 (Momany) and 4839344 (Bowers et al)

Examples of other peptides which can be used in the practice of this invention are those disclosed in US Patent No. 4589881 (30 or more amino acid residues) of Pierschbacher et al; US Patent No, 4544500 (20-30 residues) of Bidle et al; and 10 EP0204480 ( >34 residues) of Dimarchi et al and teriparatide. In some embodiments, the therapeutic agent can include a polysaccharide, such as a glyeosatninoglvean. Exemplary glycosaminoglvcans include heparin, heparin derivatives, heparan sulfate, chondroitin sulfate, dermatan sulfate, and hyaluronic acid. Examples of heparin derivatives include, but are not limited to, low molecular weight heparins such as enoxaparin, daltcpurin and 15 tinzaparin. A therapeutic agent with a heparin-like effect is fondaparinux.

Other examples of therapeutic agents include, but are not limited to hormones such as insulin, erythropoietin (EPO), glucagon-like peptide 1 (GEP-!). melanocyte stimulating hormone (alfa-MSIT). parathyroid hormone (PTH), teriparatide. growth hormone (Gl 1). leuprolide, leuprolide acetate, factor VIII. growth hormone releasing 20 hormone (GHRH), peptide YY amino acids 3-36 (PYY^,), calcitonin, somatotropin, somatostatin, somatomedin, interleukins such as inierleukin-2 (IL-2), alfa-1 -antirypsin. granulocyte/monocytc colony stimulating factor (GM-CSF), granulocyte colony stimulating factor (G-CSF), T20. testosterone, interferons such as interferon-alfa (IFN-n) ΙΕΝ-β and ΙΕΝ-γ. luieinizing-hormone (Eli), follicle-stimulating hormone (ESH). human 25 chorionic gonadotropin (hCG), enkephalin, dalargin, kyotorphin, basic fibroblast growth factor (bFGF). hirudin, hirulog, luteinizing hormone releasing hormone (LHRH). gonadotropin releasing hormone (GnRH) analog, brain-derived natriuretic peptide (BNP), tissue plasminogen activator (TPA), oxytocin, and analogs and combinations thereof.

Oltter examplcs-of therapeutic agents include, but are not limited to analgesic 30 agents, anti-migraine agents, anti-coagulant agents, anti-emetic agents, cardiovascular, anti-hypertensive and vasodilator agents, sedatives, narcotic antagonists, chelating agents, anti-diuretic agents and anti-neoplastic agents. wo 2010/032140 IO7IB2(MWAM>7155 14

Analgesics include, but are not limited to, fentanyl, sufentanil, butorphanol. buprenorphine, levorphanol, morphine, hydromorphone, hydrocodeine, oxymorphone. methadone, lidocaine, bupivacaine, diclofenac, naproxen,, paverin, and analogs thereof. Anti-migraine ugents include, but are not limited to naratriptan. nuproxen. alnioiriptan. 5 butalbital, frovatriptan, sumatriptan, rizatriptan, acetaminophen, isomethepiene. butorphanol. dichloralphenazone, ergot alkaloids such as dihydroergotamtne and ergotamine. nonsteroidal anti-inflammatory drugs (NSAIDs) such as ketoprofen and ketorolac, eletriptan. butorphanol, topiramate, zolmitriptan, caffeine, aspirin and codeine, and analogs and combinations thereof. t() Λιπί-coagulant agents include, but are not limited to heparin, hirudin, low molecular weight heparins and analogs thereof and fondaparinux. Αηΰ-cmetic agents include but am not limited to scopolamine, ondansetron, domperidone. eloclopramide, and analogs thereof. Cardiovascular, anti-hypertensive and vasodilator agents include, but are not limited to, diltiazem. clonidine, nifedipine, verapamil, tsosorbide niononitraie. 15 organic nitrates, nitroglycerine and analogs thereof. Sedatives include, but are not limited to, benzodiazeines, phenoth iozines and analogs thereof. Narcotic antagonists include, hut are not limited to, naltrexone, naloxone and analogs thereof. Chelating agents include, but are not limited to deferoxamine and analogs thereof. Anti-diuretic agents include, but are not limited to. desmopressin, vasopressin and analogs (agonists') thereof such as 20 terlipressin; the trade name of teriipressin is glypresstn ®. Anu-neoplastic agents include, but are not limited to, 5-fluorouracil, bleomycin, vincristine, procarbazine, temezolamide, 6-thioguaninc. hydroxyurea , cytarabine, cyclophosphamide, doxorubicin, vinca alkaloid, epiruhiem. etoposide, ifosfamide, carboplatin and oilier platinum based antincoplastic drugs (such as carboplatin-(ParapIatin®, tetraplarin. oxaliplatin, aroplatin and iransplatin). 25 vinblastine, vinorelbine. chlorambucil, busulfan. mechlorethaminc, mitomycin, dacarba/tne. thiotepa, daunorubicin. idarubicin. mitoxuntrone. csperamicin A1, dactinomycin, plicamyctn. carmustine. lomustine (CCNU), tauromustine, streptozocin, melphalan. dactinomycin, procarbazine, dexamethasone, prednisone. 2-chlorodeoxyadenosine, cytarabine, docetaxcl, fludiirabine, gemcitabine. hereeptin. 30 hydroxyurea. irinotecan, methotrexate, rituxin. semustine. tomudex and topotecan, taxol and tuxol-like compounds and analogs and combinations thereof. WO 2011)/03214(1 15

Additional examples of therapeutic agents include, but are not limited to coagulation factors and neurotrophic factors, anti-TNF antibodies and fragments of 'I’Nl· receptors. 'llierapcutic agents also include pharmaceutically active agents selected from the 5 group consisting οΓ vitamin B12, a bisphosphonate (e.g., distxlium pamidronaie, alendronate, etidronate, tiludronaLe, risedronate, zoledronic ucid, sodium clodronate, or ibandronic acid), taxol, caspofungin. or an aminoglycoside antibiotic. Additional therapeutic agents include a toxin, or an antipathogenic agent, such as an antibiotic (e.g. vancomycin), an antiviral, an antifungal, or an anti-parasitic agent, ‘rhe therapeutic agent 10 can itself be directly active or can be activated in situ by the composition, by a distinct substance, or by environmental conditions.

In some embodiments, the composition can include a plurality of therapeutic agents (combination drugs). For example, the composition can include Factor VIII and vWF. Cil J3-1 and PYY, lFN-α and nucleotide analogues (i.e, ribavirin), and alendronate 15 or insulin and GLP-1. in some embodiments, the composition can include a small molecule and a peptide or protein. Exemplary combinations' include a combination of IFN-n and nucleotide analogues (i.e. ribavirin) for the treatment of hepatitis C, teriparatide and alendronate for treatment of bone disorders, a combination of Gl i plus the medications 20 for l-irv therapy (e.g. I1AART) to simultaneously treat the viral infection and the accompanying l itV lipodystrophy or AIDS wasting side effects. Combinations of two small molecules can be used when one of them generally has poor absorption or bioavai lability even if the other generally has effective absorption or bioavailability, such as some antibiotics (e.g. a combination of vancomycin and an aminoglycoside such as 25 gentamicin. Exemplary combinations for the treatment and prevention of metabolic disorders such as diabetes and obesity also include combination of insulin and metformin, insulin and rosiglitazone, GLP-1 (or exenatide) and metformin, and GIJ’-I (or exenatide) and rosiglitazone,

Indications and conditions which may be treated by (bndaparinux formulated as 30 described herein include deep vein thrombosis, hip or knee replacement, and bed-hound patients.

In some embodiments of the compositions described herein, the composition includes a combination of a protein or peptide with S'mull molecules that cither do or do WO 20111/432140 PCI71B2M19/1H17155 16 mil have good absorption or bioavailability. For example. a composition can include at least one therapeutic agent that may generally be characterized as poorly absorbable or poorly bioavailable. The composition can also be used lor the administration of therapeutic agents that are absorbed in the stomach and/or intestine, but cause irritation to 5 the stomach and/or intestine and therefore are difficult to tolerate. In such a situation, a subject could benefit if The bioavailability of the therapeutic agent were enhanced or if more of the therapeutic agent were absorbed directly into the blood stream; if less therapeutic agent is administered there will clearly he less chance of causing irritation to the stomach and /or intestine. Thus compositions of the invention arc envisaged which 10 comprises therein two or more therapeutic agents.

In general, the composition may include from ahout 0.01% to about 50% by weight of the therapeutic agent e.g. about 0.01,0,02 0,05. 1,2. 3. 4. 5, 6. 7. 8. 9. 10. 15. 20. 25. 30. 35, 40, 45, or 50% by weight The maximum included in the composition is often in the range of about 6%-33% by weight of the therapeutic agent. 15 In some embodiments of the compositions described herein, the solid form including the therapeutic agent also includes a stabilizer (e.g. a stabilizer of protein structure). Stabilizers of protein structure am compounds that stabilize protein structure under aqueous or non-aqueous conditions or can reduce or prevent aggregation of the therapeutic agent, for example (luring a drying process such as lyophilization or other 20 processing step. Stabilizers of structure can be polyanionic molecules, such as phytic acid, polyvalent ions such as Ca, Zn or Mg, saccharides such as a disaedtaride (e.g,, trehalose, maltose) or an oligo or polysaccharide such as dextrin or dextran, or a sugar aicohol such as mannitol, or an amino acid such as glycine, or polycationic molecules, such as spermine, or surfactants such as Tween 80 or Span 40 or pluronic acid. 25 Uncharged polymers, such as methyl cellulose and polyvinyl alcohol, are also suitable stabilizers.

Medium chain fatty acid salt: 'Ihe compositions described herein include the salt of a medium chain fatty acid or a derivative thereof in a solid form. For example, die salt of die medium chain fatty acid 30 is in the form of a particle such as a solid panicle. In some embodiments, the panicle may lie characterized as a granulated panicle. In at least some embodiments, the solid form may generally result from a spray drying or evaporation process. In preferred embodiments, the salt of the medium chain fatty acid is in die same panicle as the WO 21)10703214« therajicutic agent. For example, the therapeutic agent and the salt of the medium chain fatty acid can be prepared together by first preparing a solution such as an aqueous solution comprising both the therapeutic agent and the salt of' the medium chain fatty acid and co-lyophilizing the solution to provide a solid form or particle that comprises both the 5 therapeutic agent and the salt of the medium chain fatty acid (and other ingredients). As described above, the resulting solid particles arc associated with a hydrophobic medium. For example, the solid particles may be suspended or immersed in a hydrophobic medium

In different embodiments of die compositions described herein the medium chain fatty acid salt may be in the same panicle or in a different panicle than that of die API. It 10 was found that bioavailability of a cargo compound was lower if die medium chain fatty acid was in a different particle than the therapeutic agent i.e. there was improved bioavailabiiity if the medium chain fauy acid salt and the cargo compound were dried after solubilization together in the hydrophilic fraction, it is believed that if the medium chain fatty acid salt and the cargo compound are dried after solubilization together in the 15 hydrophilic fraction then they are in the same particle in the Final powder.

Mediutn chain fatty acid salts include those having a carbon chain length of from about 6 to about 14 carbon atoms. Examples of fatty acid salts are sodium hexanoate, sodium heptanoate. sodium octanoate (also termed sodium caprylate), sodium nonanoate, sodium decanoate, sodium undecanoate. sodium dodecanoate. sodium Lridecanoatc, and 20 sodium tetradecanoate. In some embodiments, the medium chain fatty acid salt contains a cation selected from the group const sting of potassium, lithium, ammonium and other monovalent cations e.g. the medium chain fatty acid salt is selected from lithium octanoate or potassium ocLanoate or arginine octanoate or other monovalent salts of the medium chain fatty acids. The inventors found that raising the amount of medium chain 25 fatty acid suit increased the bioavailability of the resulting formulation. In particular. raising the amount of medium chain fatty acid salt, in particular sodium octanoate. above 10% to a range of about 12% to L5 % increased the bioavailability of the therapeutic agents in the pharmaceutical compositions described herein.

In general, the. amount of medium chain fatty acid salt in the compositions 30 described herein may be from 10% up to about 50% by weight of the bulk pharmaceutical composition. For example, the medium chain fatty acid suit may be present at an amount of about 10% -50%. preferably about 11 %>-40% most preferably about 11 %-2K% by weight for example at about 12%-13%. 13%-14%, 14%-L5% ; 1S%-16%·, 16%-17%, WO 2011)/032140 l*C'l7l820tiy/007155 18 ! 7%-18%, 18%-19%, 1998-20%. 20%-21%, 21%-22%,2 2%-23%. 23%-24%,2 498-25%·. 2598-2698. 2698-27%. or 27%-28% by weight of the bulk pharmaceutical composition. In other embodiments the medium chain fatty acid salt may be present at an amount of at least about 11%, at least aboutl2%, at least about 13%. at least abouil4%. at least about 5 15% at least about 16%,at least about 17%. at least about 18%. at least about 19%. at least about 20%, at leust about 21%, at least about 22%. at least about 23%. at least about 24%. at least about 25%=. at least ttbout 26%, at least about 27% or at least about 2898 by weight of the bulk pharmaceutical composition. In specific embodiments die medium chain fatty acid salt (sodium, potassium, lithium or ammonium salt or a mixture thereof) is present at 10 about 12% -21 % by weight of the bulk pharmaceutical composition preferably 11 %-18%> or about 11 98-17%, or 12%-16%or 1298-15% or 1348-16% or t.3%-15%, or 1498-16%· or 1498- i s% oj- 15%-16% or most preferably 15%· or 16%. in specific embodiments the medium chain fatty acid salt (having a carbon chain length of from about 6 to about 14 carbon atoms particularly 8, 9 or 10 carbon atoms) is present at. about 12% -21%· by 15 weight of the bulk pharmaceutical composition preferably 11 %-18%· about 11 %<-17% or 12%-16% or 1298-15% or 1398-16% or 1398-15% or 14%-16%· or 1498-15%. or 1598-16% or most preferably 15% or 16%·. In specific embodiments die medium chain fatty acid salt (for example sales of octanoic acid, salts of suberic acid, salts of geranic acid) is present at about 12% -21 % by weight of the hulk pharmaceutical composition preferably I I %>-18%· 20 about t ] 98-17% or I2%-16% or 1298-15%. or 13%-16%· or 1398-15% or 1498-1698 or 1498-15%) or 1.598-16% or most preferably 15%’ or 1698. In certain embodiments the medium chain fatty' acid salt is present in die solid powder at an amount of 5Q% to 90%. preferably at an amount of 70% to 80%,

One embodiment of the invention comprises a composition comprising a 25 suspension which consists essentially of an admixture of a hydrophobic medium and a solid form wherein the solid form comprises a therapeutically effective amount of a iherapeudc agent and at least one salt of a medium chain fatty acid, and wherein the medium chain fatty acid salt is not a sodium salt. The salt may be the sals of another cation e.g, lithium, potassium or ammonium; an ammonium salt is preferred. 30 Matrix forming polymer:

In certain embodiments die composition of the invention comprises a suspension which comprises an admixture of a hydrophobic medium and a solid form wherein the solid form comprises a therapeudcally effective amount of a therapeutic agent, at least WO 201 (1/032140 one sail of a medium chain fatty acid and a matrix forming polymer, and wherein the matrix forming polymer is present in the composition ai an amount of 3% or more by weight. In certain embodiments the composition comprises a suspension which consists essentially of an admixture of a hydrophobic medium and a solid form wherein the solid 5 form comprises a therapeutically effective amount of a therapeutic agent, at least one salt of a medium chain fatty acid and a matrix forming polymer, and wherein the matrix forming polymer is present in the composition at an amount of 3% or more by weight. In particular embodiments the matrix forming polymer is dextran or polyvinylpyrrolidone (PVP). In particular embodiments the polyvinylpyrrolidone is present in the composition 10 at an amount of about 2% to about 20% by weight, preferably at an amount of alxtut 3% io about 18 % by weight, more preferably at an amount of about 5% to about 15 % by weight, most preferably at an amount of about 10 % by weight. In certain particular embodiments the polyvinylpyrrolidone is PVP- 12 and/or has a molecular weight of about 3000, Other matrix forming polymers have a similar effect in the compositions of the 15 invention: such matrix forming polymers include ionic polysaccharides (for example alginic acid and alginates) or neutral polysaccharides (for example dextran and HPMO. polyacrylic acid and poly methacrylic acid derivatives and high molecular weight organic alcohols (for example polyvinyl alcohol), 20 Protease inhibitors:

It is generally accepted in the art of delivery of proteins, polypeptides and peptides that protease inhibitors normally have to be added to the formulation to prevent degradation of the API. However in the formulations of the instant invention it is not necessary to add protease inhibitors. The formulations of die invention appear to confer 25 stability of die therapeutic agent to protease degradation within the time-frame of aciixriiy i.e, the formulations of the invention are apparently environment inhibitory for enzyme activity. Additionally, die inventors performed an experiment wherein the protease inhibitor aproiinin was added to a formulation and this had no beneficial effect on activity. A similar experiment was performed where the protease inhibitor a- 30 aminocaproic acid was added to a formulation and this too had no beneficial effect on activity. Therefore. in some embodiments, a pharmaceutical composition described herein is substantially free of a protease inhibitor. WO 2tH 0/(132140 20

Hydrophilic, fraction:

In embodiments of the invention, the above compounds. including the therapeutic agent and the medium chain fatty acid salt are solubilized in an aqueous medium and then 5 dried to produce a powder, fhe drying process may be achieved for example by lyophilization or granulation. The powder obtained is termed the “hydrophilic fraction”.

In the hydrophilic fraction water is normally present at an amount of less than 6%.

Lyophilization may be carried out as shown in die Examples herein and by methods known in the art e.g. as described in Lyophilization: Introduction.and Hasic 10 Principles . Thomas Jennings, published by Interphanu/CRC Press Ltd (1999. 20(12) Die lyophilizate may optionally be milled ( e.g.-below 150 micron) or ground in a mortar. During industrial production Ute lyophilizate is preferably milled before mixing of the hydrophilic fraction and the hydrophobic medium in order to produce bateb-io-batch re product bi lily. 15 Granulation tnay be carried out as shown in the Examples herein and by methods known in the art e.g. as described in Granulation. Salman et al , eds, Elsevier (2006) and in Handbook of Pharmaceutical Granulation Technology. 2nd edition, Dilip M, Parikh, ed. (2005

Various binders may be used in the granulation process such as celluloses 20 (including microcrystalline celluloses), lactoses (e.g. lactose monohydrate), dextroses, starch and mannitol and other binders as described in die previous two nefemnees.

Hydrophobic Medium:

Oil: As described above, in the compositions of the invention described herein the 25 therapeutic agent and the medium chain fatty acid salt are in intimate contact or association with a hydrophobic medium. For example, one or both may be coated, suspended, immersed or otherwise in association wtdi a hydrophobic medium. Suitable hydrophobic mediums can contain, for example, aliphatic, cyclic or aromatic molecules. Examples of a suitable aliphatic hydrophobic medium include, but are not limited to. 30 mineral oil. fatty acid monoglycerides, diglycerides, triglycerides, ethers, esters, and combinations thereof, Examples of a suitable fatty acid are octanoic acid, dccanoic acid and dodecanoic acid, also C7 and C9 fatty acids and cli-acidic acids such as sehacic acid, and suberic acid, and derivatives thereof. Examples of triglycerides include, but arc not 21 iintited to.long chain triglycerides, medium chain triglycerides, and short chain triglycerides. For example, die long chain triglyceride can be castor oil or coconut oil or olive oil. and the short chain triglyceride can he glyceryl tributyrate and the medium chain triglyceride cun be glyceryl tricaprylatc. Monoglycerides are considered to be 5 surfactants and are described below. Exemplary esters include ethyl iso valerate and butyl acetate. Examples of a suitable cyclic hydrophobic medium include, but are not limited to. terpenoids, cholesterol, cholesterol derivatives (e.g. cholesterol sulfate), and cholesterol esters of fatly acids. A non-limiting example of an aromatic hydrophobic medium includes benzyl benzoate. 10 In some embodiments of the compositions described herein, it is desirable that the hydrophobic medium include a plurality of hydrophobic molecules. In some embodiments of the compositions described herein die hydrophobic medium also includes one or more surfactants (see below).

In some embodiments of die compositions described herein, the hydrophobic 15 medium also includes one or more adhesive polymers such as methylcellulose. ethylcell ulose. hydroxypropyhnethylcellulose (ILPMC), or poly(acrylate) derivative Carbopol@934P (C934P). Such adhesive polymers may assist in the consolidation of the formulation and/or help its adherence to mucosal surfaces.

Surface Active Agents (surfactants): The compositions of this invention described 20 herein can further include a surface active agent. For example, the surface active agent can he a component of die hydrophobic medium as described above, und/or the surface active agent can he a component of a solid form as described above, for example in the solid form or particle that includes the therapeutic agent.

Suitable surface active agents include ionic and non-ionic surfactants. Examples 25 of ionic surfactants arc lecithin (phosphatidyl choline), bile salts and detergents.

Examples of non-ionic surfactants include monoglycerides, cremophore. a polyethylene glycol fatty alcohol ether, a sorbitan fatty acid ester, a polvoxyediylene sorbitan fatty acid ester. Solutol I IS 15, or a poloxamer or a combination thereof. Examples of monoglycerides are glyceryl monocaprylate (also termed glyceryl monooctuiioate). 30 glyceryl monodeeanoate, glyceryl monolaurate, glyceryl monotnyri.state. glyceryl monosiearate, glyceryl nionopalmitate, and glyceryl monooleate. Examples of sorbitan faily acid esters include sorbitan monolaurate, sorbitan monooleate. and sorbitan monopahnitate (Span 40), or a combination thereof. Examples of polyoxyethylene WO 2010/032140 PC171B2009/007155 22 sorbitan fatty acid esters include polyoxyethylene sorbitan ntonooieate (Tween 80), polyoxyethylene sorbitan tnonostearaie. polyoxyethylene sorbitan monopalmiiate or a combination thereof. The commercial preparations of monoglvcerides that were used also contain various amounts of diglyecrides and triglycerides. 5 Compositions described herein including a surface active agent generally include less than about 12% by weight of total surface active agent (e.g. less than about 10%., less than about 8%, less than about 6%. less than about 4%, Jess than about 2%. or less than about 1%). In particular embodiments of the invention the total sum of all the surfactants is about 6%. 10 Methods of making pharmaceutical compositions and the compositions produced:

Also included in the invention are methods of producing the compositions described herein. dhus one embodiment of the invention is a process for producing a pharmaceutical composition which comprises preparing a water-soluble composition comprising a therapeutically effective amounL of at least one therapeutic agent and a 15 medium chain fatty acid salt (as described above), drying the water soluble composition to obtain a solid powder, and suspending die solid powder in a hydrophobic medium, to produce a suspension containing in solid form the therapeutic agent and the medium chain fatty acid salt, thereby producing die pharmaceutical composition, wherein the pharmaceutical composition contains 10% or mom by weight of medium chain fatty acid 20 salt.

One embodiment is a process for producing a pharmaceutical composition which comprises providing a solid powder of a dierapeutically effective amount of at least one therapeutic agent and a solid powder comprising a medium chain fatty acid salt, and suspending the solid powders in a hydrophobic medium, to produce a suspension 25 containing in solid form die therapeutic agent and the medium chain fatty acid salt, thereby producing the pharmaceutical composition, wherein the pharmaceutical composition contains 10% or more by weight of medium chain fatty acid salt.

In tme embodiment of die processes and compositions described herein, the water-soluble composition is an aqueous solution. In certain embodiments the diving of the 30 water-soluhle composition is achieved by lyophilization or by granulation. In the granulation process a binder may be added to the water soluble composition before drying. In certain embodiments die drying step removes sufficient water so that the water content in the pharmaceutical composition is lower than about 6% by weight, about 5% by weight, about 4% by weight, about 3% or about 2 % or about 1% by weight. In certain embodiments of the processes and compositions described herein the drying step removes an amount of water so that the water content in the solid powder is lower than 6% or 5% or 4% or 3% or preferably lower than 2% by weighL The water content is normally low 5 and the water may be adsorbed to the solid phase during lyophilization i.e. the water may be retained by imennoleeular bonds. In certain embodiments the water soluble composition additionally comprises a stahilizer for example methyl cellulose. In preferred embodiments' of the of die processes und compositions described herein die hydrophobic medium is castor oil or glyceryl tricaprylate or glyceryl tribuiyraie or a combination 10 thereof and may additionally contain octanoic acid; in certain embodiments the hydrophobic medium comprises an aliphatic, oleftnic. cyclic or uromatic compound, a mineral oil, a paraffin, a fatty acid such as octanoic acid, a monoglyceride, a diglyceride. a triglyceride, an ether or an ester, ora combination thereof. In certain embodiments of the processes and compositions described herein the triglyceride is a long chain 15 triglyceride, a medium chain triglyceride preferably glyceryl tricaprylaie or a short chain triglyceride preferably glyceryl tribuiyraie, and the long chain triglyceride is castor oil or coconut oil or a combination thereof. In certain embodiments of the processes and compositions described herein the hydrophobic medium comprises custor oil or glyceryl iricaprylate or glyceryl tributyrate or a combination or mixture thereof, and may 20 additionally comprise octanoic acid. In certain embodiments of the processes and compositions described herein the hydrophobic medium comprises glyceryl Lricaprylate or a low molecular weight ester for example ethyl isovalerate or butyl acetate, tn certain embodiments of the processes and compositions described herein die main component by weight of the hydrophobic medium is castor oil and may-additionally comprise glyceryl 25 lricaprylate. In certain embodiments of the processes and. compositions described herein the main component by weight of the hydrophobic medium is glyceryl lricaprylate and may additionally comprise castor oil. A basic formulation is provided as an embodiment wherein the hydrophobic medium consists essentially of castor oil, glyceryl tnonooleaie and glyceryl tributyrate; in 30 a further embodiment of the basic formulation the hydrophilic fraction consists essentially of therapeutic agent, PVP-12 and sodium octanoate. Λ particular formulation is provided as an embodiment wherein the hydrophobic medium consists essentially of glyceryl tricaprylaie, castor oil, glyceryl monocapryiaie,

anti Tween 80. anti the hydrophilic fraction consists essentially of iherapeutic agent ( e.g. octreotide), PVP-12 and sodium octanoate. Another particular formulation is provided as an embodiment wherein the hydrophobic medium comprises glyceryl trieaprylate. castor oil, glyceryl inonocaprylaie, and Tween 80, and the hydrophilic fraction comprises therapeutic agent (e.g. octreotide), PVP-12 and sodium octanoate. In certain embodiments the hydrophobic medium consists essentially of glyceryl trieaprylate and in certain embodiments additionally contains castor oil and /or glyceryl monocaprylate. in certain embodiments the composition comprises a suspension which consists essentially of an admixture of a hydrophobic medium and a solid form wherein the solid form comprises a therapeutically effective amount of a therapeutic agent and at least one salt of a medium chain fatty acid, and wherein the medium chain fatty acid salt is present in the composition at an amount of 10% or more by weight. In certain embodiments the hydrophobic medium consists essentially of castor oil. glyceryl monooieate and glyceryl tributyrate; or the hydrophobic medium consists essentially of glyceryl trieaprylate and glyceryl monocaprylate; or the hydrophobic medium consists essentially of castor oil. glyceryl σι caprylate and glyceryl monocaprylate. In certain embodiments the hydrophobic medium comprises a triglyceride and a monoglyceride and tn certain particular embodiments the monoglyceride has the same fatty acid radical as the triglyceride. In certain of these embodiments the triglyceride is glyceryl iricaprylaie and tire monoglyceride is glyceryl monocaprylate. In certain embodiments the medium chain fatty acid salt in the water-soluble composition has the same fatty acid radical as the medium chain monoglyceride or as the medium chain triglyceride or a combination thereof. In certain of these embodiments the medium chain fatty acid salt is sodium caprylate (sodium-octanoate)-and the-monoglyceride is glyceryl.monocaprylate and the triglyceride is glyceryl tricapiyiaie.

Many of the compositions described herein comprise a suspension which comprises an admixture of a hydrophobic medium and a solid form wherein the solid form comprises a therapeutically effective amount of a therapeutic agent and at least one salt of a medium chain fatty acid, and wherein the medium chain fatty acid salt is present in the composition al an amount of 10% or more by weight The solid form may be a panicle (e.g. consist essentially of particles, or consists of particles 1, The panicle may be produced by lyophilization or by granulation.

In a particular embodiment die formulation consists essentially of a suspension which comprises an admixture of a hydrophobic medium and a solid form wherein the solid Form comprises a therapeutically effective amount of a therapeutic agent and about 10-2(1% preferably 15% medium chain fatty acid salt preferably sodium octanouLe. and 5 ubout 5- 10% preferably 10% PVP- .1.2: and wherein the hydrophobic medium comprises about 20-80%- , preferably 30-70% triglyceride preferably glyceryl tricaprylate or glyceryl tributyrate or castor oil or a mixture thereof, about 3-10% surfactants, preferably about 6%. preferably glyceryl monocaprvlate and Tween 80 and about 1% water; in particular embodiments the therapeutic agent is present at an amount of less than 33%. or less than 10 25%, or less than 10%, or less than 1 % or less than 0.1 % . JTie solid form may be u panicle (e.g. consist essentially of particles, or consists of panicles). ]"hc particle may be produced bv lyophilization or by granulation. In a particular embodiment the solid form may he a particle and may he produced by lyophilization or by granulation,

In a Further embodiment the formulation consists essentially of a suspension 15 which comprises an admixture of a hydrophobic medium and a solid form wherein the solid form comprises a therapeutically effective amount of a therapeutic agent and about 10-20% preferably 1.5% medium chain fatty acid salt preferably sodium octanoate and about 5- 10% preferably 10% PVP- 12; and wherein the hydrophohic medium comprises about 20-80% , preferably 30-70% medium or shun chain triglyceride preferably glyceryl 20 tricaprylate. or glyceryl tributyrate, about 0- 50% preferably 0-30%. castor oil. about 3-10%/ surfactants, preferably about 6%), preferably glyceryl monocaprylaic and Tween 80.and about 1 % water, in particular embodiments the therapeutic agent is present at an amount of less than 33%, or less than 25%. or less than 10%. or less than 1% or less than 0.1% . 25 Ina particular embodiment the formulation consists essentially of a suspension which comprises an admixture of a hydrophobic medium and a solid form wherein the solid form comprises a therapeutically effective amount of a therapeutic agent and about 15% sodium octanoate and about 10% PVP- 12; and wherein the hydrophobic medium comprises about 41% glyceryl tricaprylate. about 27% castor oil. about 4%’ glyceryl 30 monocaprylate, about 2% Tween 80, about 1 % water and 1 % or less therapeutic agent ; when the therapeutic agent is octreotide it is present at about 0.058%).

In another particular embodiment the formulation consists essentially a suspension which comprises an admixture of a hydrophobic medium and a solid form WO 201(1/032140 H071B2009/007155 26 wherein ihe solid form comprises a therapeutically effective amount of a thenipcuLic agent and about 15% sodium octanoate and about 10% PVP- 12: and wherein the hydrophobic medium comprises about 68% glyceryl tricapryltne. about 4% glyceryl monocaprylutc. about 2% Tween 80. about 15% sodium octanoate. about 10% PVP- 12, 5 about 1 % water and less than 1 %- therapeutic agent; when the therapeutic agent is octreotide it is present at about 0.058%'.

One embodiment is a composition comprising a suspension which comprises an admixture of a hydrophobic medium and a solid form wherein the solid form comprises a therapeutically effective amount of octreotide and at least one salt of a medium chain 10 Fatty acid: in a Further embodiment the medium chain fatty acid salt is present in the composition at an amount of 10% or more by weight, preferably 15% by weight; in a further embodiment the solid form additionally comprises a matrix-forming polymer. In a further embodiment the matrix forming polymer is dextran or polyvinylpyrrolidone (1 ’VP). In a specific embodiment the.matrix forming polymer is polyvinylpyrrolidone and 15 the polyvinylpyrrolidone is present in the composition at an amount of about 2% to about 20%; by weight, preferably about 10 % by weight tn a specific embodiment the polyvinylpyrrolidone is PVP- 12 and /or the polyvinylpyrrolidone has a molecular weight (4’about 3000. In specific embodiments the hydrophobic medium consists essentially of glyceryl tricaprylate and the solid form additionally consists of PVP-12 and sodium 20 octanoate. In more specific embodiments the hydrophobic medium additionally consists of castor oil or giyceryl nionocaprylatc or a combination thereof and a surfactant in further specific embodiments the hydrophobic medium consists of glyceryl tricaprylatc. glyceryl monocaprylate, and polyoxyethylene sorbitan monooleatc (Tween 801. in a further embodiment the solid fonn consists essentially of octreotide. PVP-12 and sodium 25 octanoate. In a particular embodiment the composition contains about 41 % of glyceryl tri caprylate, about 27% castor oil, about 4% glyceryl i non ocapry late, about 2% '['ween 80. ahout 15% sodium octanoate. about 10% PVP- 12, about 1%’ water and about 0.058%; octreotide. In another particular embodiment the composition contains ahout 68% of glyceryl tricaprylatc. about 4% glyceryl tnonocaprylate, about 2% Tween 80. about 15% 30 sodium octanoate, about 10% PVP-12, about 1% water and about 0.058%» octreotide.

In all the above formulations, the percentages recited are weiglu/weight and the solid form may be a panicle (e.g. consist essentially of particles, or consists of particles). The particles may he produced by lyophilization or by granulation. WO 21)111/032141) PC'I7lB2d 1)9/007155 27

Under normal storage conditions, liie therapeutic agent within the formulations of the invention is stable over an extended period of time. The chemical and physical state of the formulation is stable. Once administered to the intestine the therapeutic agent is protected from damage by the Gl environment since the formulations are oil-based and 5 therefore a separate local environment is created in Lhe intestine where the therapeutic agent is contained in oil droplets, which confers stability in vivo.

In certain embodiments the process produces a composition which consists essentially of a therapeutic agent and a medium chain fatty acid sal) and a hydrophobic medium. In embodiments of the invention the solid powder (solid form) consists 10 essentially of a therapeutic agent and a medium chain fatly acid salt. Further embodiments of the invention are pharmaceutical compositions produced by the process describe herein. In certain pharmaceutical compositions the therapeutic agent is a protein, a polypeptide, a peptide, a glycosaminoglycan, a polysaccharide, a small molecule or a polynucleotide and in particular embodiments die dierapeutic agent is insulin, growth 15 hormone, parathyroid hormone, teriparatide, interferon-alfa (IfTM-a). a low molecular weight heparin, leuprolide, fondaparinux, octreotide, exenatidc. lerltpressin. vancomycin or gentamicin. Particular embodiments of llte invention comprise an oral dosage form comprising die pharmaceutical composition, in particular an oral dosage form which is enteric coaled. Further embodiments of die invention comprise a capsule containing the 20 compositions of the invention, and in various embodiments the capsule is a hard gel or a soft gel capsule, and generally lhe capsule is enteric-coaled. Other embodiments of the invention comprise a rectal dosage form comprising the pharmaceutical composition, in particular a suppository, or a buccal dosage form. A kit comprising instructions and die dosage form is also envisaged. 25 The therapeutic agent or medium chain fatty acid salt, or any combination of therapeutic agent and other components, such as protein stabilizers, can be prepared in a solution of a mixture (e.g. forming an aqueous solution or mixture) which can be lyophilized together and then suspended in a hydrophobic medium. Other components of die composition can also be optionally lyophilized or added during reconstitution of the 30 solid materials.

In some embodiments, die therapeutic agent is solubilized in a mixture, for example, including one or more additional components such as a medium chain fatty acid salt, a stabilizer and/or a surface active agent, and the solvent is removed to provide a WO 2010/03214» PCr/lB2000/007155 28 resulting solid powder (solid fonn), which is suspended in a hydrophobic medium. In some embodiments, the therapeutic agent and/or the medium chain fatty acid salt may be Ibnned into a granulated panicle that is then associated with the hydrophohic medium (for example suspended in the hydrophobic medium or coated with the hydrophobic 5 medium). In general, the compositions described herein are substantially free of 'membrane fluidizing agents" such as medium chain alcohols. “Membrane fluidizing agents" are defined as medium chain alcohols which have a carbon chain length of from 4 to 15 carbon atoms (e.g., including 5 to 15, 5 to 12. 6.‘ 7, 8. 9, 10, or 11 carbon atoms). For example, a membrane fluidizing agent can he a linear 10 (e.g. saturated or unsuturated). branched (e.g. saturated or unsaturaied). cyclical (e.g. saturated or unsaturaied), or aromatic alcohol. Examples of suitable linear alcohols include, but are not limited to, butanol, pentanol, hexanol, heptanol, octanol. nonanol. decanol, undecanol. dodecannl, tridecanol, tetradecanol, and pcntadecunol. Examples of branched alcohols include, but are not limited to, geraniol, farnesol. rhodinol. citronellol. 15 An example of a cyclical alcohol includes, but is not limited to, menthol, terpineol, myrtenol. perillvl and alcohol. Examples of suitable aromatic alcohols include, but are not limited to. benzyl alcohol, 4-hydroxy cinnamic acid, thymol, styrene glycol, and phenolic compounds. Examples of phenolic compounds include, but are not limited to, phenol, nt-cresol. and m-chlorocresol. 20 If desired, the pharmaceutical composition may also contain minor amounts of non-toxic auxiliary substances such pH buffering agents, and other substances such as for example, sodium acetate and triethanolamine oleate.

In at least one embodiment, a therapeutic agent, such as a protein, may be chemically modified to enhance its half-life in circulation. For example, the tlierapeutic 25 agent may undergo a process such as pegylation:

In some embodiments the process for producing a pharmaceutical composition comprises preparing a water-soluble composition comprising a therapeutically effective amount of at least one therapeutic agent and a medium chain fatty acid salt, drying the water soluble composition to obtain a solid powder, and dissolving the solid powder in a 30 solution consisting essentially of octanoic acid, thereby producing the pharmaceutical composition, which is a solution. In some embodiments, the solid fonn may he a particle (e.g., consist essentially of particles, or consists of particles), in some embodiments. the WO 2<MW»32H0 I>CT/1B2IHW/M7155 29 particle may be produced by Jyophilizaiion or by granulation. In some embodiments of ibis process ihc ocianoic acid is present in the composition at a level of about 60 % to about 90% or at a level of about 70 to about 85% preferably about 78%. In some embodiments of this process the fatty acid salt is sodium octanoatc; in further 5 embodiments of this process the medium chain fatty acid salt is present in the coniposilion al an amount of about 11%’ to about 40% by weight or at an amount of about 11 % to about 28% by weight or at an amount of about 15% by weight. In some embodiments of this process the composition additionally comprises a matrix forming polymer and in particular embodiments of this process the matrix forming polymer is 10 dextran or polyvinylpyrrolidone (PVP); in further embodiments of this process the polyvinylpyrrolidone is present in die composition at an amount of about 2% to about 20% by weight or at an amount of about. 5% to about 15 % by weight, preferably at an amount of about 10 % by weight. In certain embodiments of this process (he polyvinylpyrrolidone is PVP- 12 and /or has a molecular weight of about 3000. The 15 composition may in addition include surfactants as described above. 'ITte pharmaceutical products of these processes :ire further embodiments of the invention e.g. » composition containing ocianoic acid at a level of about 60 % to about 90%» or at a level of about 70 to about 85% preferably about 78%;. fatty acid salt, preferably sodium octanoate. present in the composition titan amount of about 11%; to about 40% by weight or at an amount of 20 about 11 % to about 28% by weight or at an amount of about 15% by weight; matrix forming polymer e.g. polyvinylpyrrolidone, preferably PVP-12, present in die composition at an amount of about 2% to about 20% by weight or preferably an amount of about 5% to about 15 % by weight, preferably at an amount of about 10 % by weight; and surfactants as described above. Ihcrc also may be small quantities of other 25 hydrophobic constituents as described above.

Capsules: Preferred pharmaceutical compositions are oral dosage forms or suppositories. Exemplary dosage forms include gelatin or vegetarian capsules like starch hydroxylpropyl-meihylcellulose ("MPMC") capsules, enteric coated, containing the bulk drug product. Capsules which may be used to encapsulate die compositions of diis 30 invention arc known in the an and are described for example in Pharmaceutical Co/wr/ex edited by Podczech and Jones. Pharmaceutical Press (2004) and in Hurd gelatin capsules today - and tomorrow, 2nd edition. Steggeman cd published by Capsugel Library (20021. WO 2010/032140 PCl71B200y/007l55 30

Additional formulations; 'ITte compositions of the invention may he formulated using additional methods known in the art. for example as described in the following publications: Pharmaceutical Dosage Forms Vols 1-3 ed. Lieberman. Laehman and 5 Schwartz, published by Marcel Dekker Inc. New York(1989): Water-insoluble Drug Formulation 2nd edition. Liu, editor, published by CRC Press, Taylor und Francis Group (2008): Therapeutic Peptides and Proteins: Formulation, Processing and Delivery Svstems. 2nd edition by Ajay K. Banga (author) published by CRC Press . Taylor and Francis Group (2006): Protein Formulation and Delivery', 2nd edition, McNally and 10 1 -lasted eds . published by Infomia Healthcare USA Inc(2008): and Advanced Drug

Formulation to Optimize Therapeutic Outcomes, Williams et al eds, published by Inlbnna Healthcare USA (2008). 1'he compositions of the invention may be formulated using microparticulaie technology for example as described in Micropart ten late Oral Drug Delivery, Gerbre- 15 Selassie ed. published by Marcel Dekker Inc (1994) and in Dey et al. Multiparticulate Drug Delivery' Systems for Controlled Release, Tropical Journal of Pharmaceutical Research. September 2008: 7 (3): 1067-1075.

Methods of treatment: The compositions described herein exhibit effective, enteral 20 delivery of an unaltered biologically active substance (Le. a therapeutic agent) and thus, have many uses. For example, the compositions described herein can be used in the treatment of diabetes.

In particular, insulin to treat and prevent subjects (patients) suffering from Type 11 diabetes (prophylaxis of diabetes), and to treat patients suffering from dysglycemia. pre- 25 diabetes and metabolic syndrome and other conditions, may be administered in

accordance with one or more embodiments of the invention. Metabolic syndrome is a combination of medical disorders that increase the risk of developing cardiovascular disease and diabetes. Metabolic syndrome is a composite of different symptoms: (1) fasting hyperglycemia (insulin resistance, type II diabetes, etc); (2) decreased HDL 30 cholesterol: (3) elevated triglycerides; (4) hi git blood pressure: (5) central obesity; and (6) proinflammaLorv stale.

One embodiment of die invention is a method of treatment or prevention of a subject suffering from the above conditions where the amount of insulin sufficient io treat WO 2(110/03214(1 PC'l71B200y/007155 31 the condition is a low dose of insulin formulated within die compositions of die invention. Low dose insulin is provided by less dian 300 or less dian 200 Units per capsule e.g 40-200 Unibs per capsule.

Terlipressin ( or odier vasopressin analogs) to Lreat subjects (patients) suffering 5 from hepaio-renal syndrome (HRS), including HRS 1 and IL bleeding esophageal varices, portal hypertension and other conditions may be administered in accordance with one or more embodiments of the invention. Such terlipressin formulations may also be used for primary and secondary prophylaxis of variceal bleeding, A composition of the invention comprises a suspension which comprises an admixture of a hydrophobic medium and a 10 solid fonn wherein die solid form comprises a therapeutically effective amount of terlipressin (or other vasopressin analogues) and at least one salt of a medium chain fatly acid,

Exenatide to improve glycemic control in subjects suffering from Type It diabetes and to treat other conditions such as obesity and for use in weight management may be 15 administered in accordance with one or more embodiments of the invention. tnierferon-alfa for the treatment of subjects suffering from chronic hepatitis C and chronic hepatitis H and to treat other conditions including cancer may be administered in accordance with one or more embodiments of the invention.

Copaxone to treat subjects suffering from multiple sclerosis and to treat tidier 2() conditions including inflammatory' diseases may be administered in accordance with one or more embodiments of the invention.

Desmopressinjo treat subjects suffering From primury nocturnal enuresis, central diabetes insipidus (DI) or bleeding disorders (Von Willebrand Disease and Hemopilia A) may tie administered in accordance with one or more embodiments of the invention. Oral 25 desmopressin preparations known in the art suffer from extremely lew oral bioa vailahi li ly.

Octreotide was first synthesized in 1979. and is an ociapeptide dial mimics natural somatostatin pharmacologically, though it is a more potent inhibitor of growth hormone, glucagon and insulin Ilian the natural hormone. Octreotide or odier analogs of 30 somatostatin may he administered in accordance with one or more embodiments of the invention for use in treating or preventing a disease or disorder in a subject suffering from a disorder such as acromegaly, abnormal Gl motility, flushing episodes associated with carcinoid syndrome, portal hypertension, an endocrine tumor (such as carcinoids, WO 21)11)/03214() PC17IB2O09/007155 VIPoma). gastroparesis, diarrhea, pancreatic leak or a pancreatic pseudo-cyst. The diarrhea may result From radiotherapy or may occur for example in subjects with vasoactive intestinal peptide-sec reting tumors (VlPomas). In addition, patients that undergo pancreatic surgery may suffer from secretion of extrinsic pancreas und are 5 vulnerable to developing pancreatic leak or pseudo-cysts which may be treated by octreotide products of the invention. Some preferred embodiments are directed to a method of treating a subject having a disorder such as acromegaly, abnormal Gl motility, Hushing episodes associated with carcinoid syndrome, portal hypertension, an endocrine tumor (such as carcinoids, VIPotna), gastroparesis, diarrhea, pancreatic leak or a 10 pancreatic pseudo-cyst which comprises administering to the subject a composition of the invention, wherein the therapeutic agent is ocueohde. in an amount sufficient to treat the disorder. Octreotide formulations of die invention may also be used for primary and secondary prophylaxis of variceal bleeding, which may be caused by portal hypertension: the varices may be gastric or esophageal. Other uses of octreotide formulations of die f 5 invention are in treatment of shock of hypovolemic (e.g. hemorrhagic) or vasodilatory (e.g. septic) origin, hepatorenal syndrome (HRS), cardiopulmonary resuscitation and anesthesia-induced hypotension. Odier analogs of somatostatin may be used in the methods and compositions in which octreotide is used,

Vancomycin (molecular weight 1449 Da ) is a glycopeptide antibiotic used in the 20 prophylaxis and treatment of infections caused by Grant-positive bacteria. The original indication for vancomycin was for the treatment of methycilin-resistant Staphylococcus aureus (MRSA). Vancomycin never became first line treatment for Staphylococcus aureus, one reason being that vancomycin must be given intravenously. Hie prior an preparations of vancomycin need to be given intravenously for systemic therapy, since 25 vancomycin does not cross through the intestinal lining. It is a large hydrophilic molecule which partitions poorly across the gastrointestinal mucosa. The only indication for oral vancomycin therapy is in the treatment of pseudomembranous colitis where it must be given orally to reach the site of infection in the colon, Vancomycin for use in treating or preventing infection in a subject may be administered orally to the subject in accordance 30 with one or more embodiments of die invention, Some preferred embodiments of the invention are directed lo a method of treating or preventing an infection in a subject which comprises administering to die subject a composition of die invention, wherein the therapeutic agent is vancomycin, in an amount sufficient to treat or prevent the infection.

Gentamicin (molecular weight = 478) is an aminoglycoside antibiotic, used to treat many types of bacterial infections, particularly those caused by gram-negative bacteria. When gentamicin is given orally in the prior art formulations, it is not systemieally active. This is because it is not absorbed to any appreciable extent from the 5 small intestine.

In addition, compositions of the invention also can be used io treat conditions resulting from atherosclerosis and the formation of thrombi and emboli such its myocardial infarction and cerebrovascular accidents. Specifically, the compositions can be used to deliver heparin or low molecular weight heparin or fondaparinux across the )0 mucosal epithelia, 'Hie compositions of this invention can also be used to treat hematological diseases and deficiency states such as anemia and hypoxia that are amenable to administration of hematological growth factors. The compositions of the invention can he used to deliver vitamin B12 in a subject at high bioavailability wherein the mucosal 15 epithelia of the subject lacks sufficient intrinsic factor. G-CSF may also be administered in accordance with various embodiments. Additionally, the compositions of this invention can be used to treat osteoporosis, such its through enteral administration of PTH. teriparatide or calcitonin once or twice or more daily.

Human growth hormone (hGl-l) to treat growth hormone deficiency in particular 20 in children may be administered in accordance with one or more embodiments, in some preferred embodiments, a composition described herein comprising growth hormone can be administered to a subject to treat or prevent metabolic and lipid-related disorders, e.g., obesity, abdominal obesity. hyperlipidemia or hvpercholestrolemia. For example a composition of the invention comprising growthhormone can be administered orally to a 25 subject thereby treating obesity (e.g., abdominal obesity). In some preferred embodiments, a composition described herein comprising growth hormone is administered to a subject to treat or prevent HIV lipodistrophy (AIDS wasting) or to treat Prader-Willi syndrome, growth disturbance due to insufficient secretion of' growth hormone ( e.g, associated with gonadal dysgenesis or Turner syndrome) , growth 30 disturbance in prepubertal children wiLh chronic renai insufficiency, and as replacement therapy in adults with pronounced growth hormone deficiency. Compositions of the invention comprising growth hormone can be administered orally to a subject to promote wound healing and attenuate catabolic responses In severe burns, sepsis, multiple trauma, WO 2010/(13214() major operations, acute pancreatitis and intestinal fistula. Many other conditions besides till deficiency cause poor growth, but growth benefits (height gains) are often poorer than when GH deficiency is treated. Examples of other causes of shortness which may be treated with compositions of the invemion comprising growth hormone are intrauterine 5 growth retardation, and severe idiopathic short stature. Other potential uses of compositions of the invention comprising growth hormone include treatment to reverse or prevent effects of aging in older adults, to aid muscle-bull di ng and as treatment for fibromyalgia.

Some preferred embodiments are directed to a method of treating a disorder such t() as obesity, i 11V lipodtstrophy. metabolic disorder, or growth deficiency in a subject which comprises administering to the subject a composition of the invention wherein the therapeutic agent (the effector) is growth hormone, in an amount sufficient to Lreat the disorder.

Some preferred embodiments are directed to a method of treating a bone disorder 15 in a subject which comprises administering to the subject a composition of the invention, wherein the therapeutic agent is teriparaLide or parathyroid hormone, in an amount sufficient to treat (he bone disorder.

Some preferred embodiments are directed to a method of treating or preventing a blood coagulative disorder in a subject which comprises administering to the subject a 20 composition of the invention wherein the therapeutic agent is heparin or a heparin derivative or fondaparinux, in an amount sufficient to treat or prevent the blood coagulative disorder. I.Aiuprolide (GnRH agonist) formulated in an embodiment of the invention may be delivered for treatment of female-infertility (c;g. once or twice daily dosage), prostate 25 cancer and Alzheimer's disease.

One embodiment of the invention relates to a method of treating a subject suffering from a disease or disorder which comprises administering to the subject a composition of the invention in an amount sufficient to treat the condition. Another embodiment of the invention relates to compositions of the invention for use in treating a 30 disease or disorder in a subject Another embodiment of the invention relates to the use of a therapeutic agent in the manufacture of a medicament by lire process of the invention for the treatment of a disorder. WO 2010/032140 PCI7IB2009/007155 35

The dosage regimen utilizing die compounds is selected in accordance with a variety of factors including type, species, age. weight, sex and medical condition of the patient; the severity of the condition to be treated; the route of administration: the renal and hepatic function of the patient; and the particular compound or salt thereof employed. 5 An ordinarily skilled physician or veterinarian can readily determine and prescribe die effective amount of the drug required to prevent, counter or arrest the progress of the condition. Oral dosages of the present invention, when used for the indicated effects, may he provided in the form of capsules containing 0.001. 0.0025,0.005. 0.01.0.025. 0,05. 0.1.0.25. 0.5. 1.0. 2.5, 5.0, 10.0. 15.0, 25-0. 50.0 or 100. 200. 300. 4()0. 500. 600, 700. 10 K(K) or 1000 mg of therapeutic agent.

Compounds of the present invention may be administered in a single daily dose. or the total daily dosage may be administered in divided doses of two. three, four, five or six times daily. In some embodiments, the composition is administered at a daily dose of' front about 0.01 to about 5000 mg/day, e.g., administered once daily (e.g,, in the morning 15 or before bedtime) or twice or more daily (e.g. in the morning and before bedtime). A representative product of the invention is an API- based formulation orally administered as enteric coaled-capsules: each capsule contains API co-lvophilizcd with PVP-12 and sodium octanoate. and suspended in a hydrophobic (lipophilic) medium containing: glyceryl tricaprylaie. glyceryl monocapryiaie. and Tween 80: in another 20 representative product of the invention castor oil is additionally present. The compositions described herein can be administered to a subject i.e. a human or an animal, in order to treat the subject with a pharmacologically or therapeutically effective amount of a therapeutic agent described herein. The animal may be a mammal e.g. a mouse, rat, pig horse, cow or sheep. As used herein the term ’'pharmacologically or therapeutically 25 effective amount" means that amount of a drug or pharmaceutical agent (the therapeutic agent) that will elicit the biological or medical response of a tissue, system, animal or human that is being sought by u researcher or clinician,

Ihe formulations of the invention allow incorporation of die therapeutic agent into the formulation without any chemical modification of the therapeutic agent. 30 Additionally, as shown above, many different therapeutic agents have been successfully Formulated within the formulations of the invention, including polypeptides, nucleotides, small molecules and even medium size proteins. Furthermore, the formulations of the invention allow for high flexibility in loading of the therapeutic agent. Loading capacity WO 2010/032140 J*CT/lB200y/007155 36 is dependent on the therapeutic agent. To dale, loading capacity limits have not been reached; however loading of up to 1.5 % wt/wt (polypeptides) and 6 % wt/wt (small molecules) has been achieved and higher loading up to 33% is envisaged. Finally, the formulations of the invention protect the cargo compounds from inactivation in die Gl 5 environment due to for example proteolytic degradation and oxidation.

[Tie function and advantages of these and other embodiments will be more fully understood from the following examples. These examples are intended to be illustrative in nature and are not to be considered as limiting the scope of the systems and methods discussed herein. WO 2010/)132140 PCt7IB2009/007155 37

EXAMPLES

Example I: Formulations A. Composition of an insulin formulation

Table 1Λ presents an example of a composition in accordance with one or more embodiments. More specifically, this composition is an insulin formulation. Insulin was obtained from Diosynth Biotechnology; sodium octanoate and NaOil from Merck; MgCh. MC400. Spun40, lecithin and castor oil from Spectrum; PVP-12 from BASF; ethyl iso valerate from Merck/Sigma; glyceryl tributyrate from Acros/Penta; and glycerol monooieate from Abitcc Corp.

Table 1A

Ingredient % w/w Hydrophilic Fraction Insulin 0.417 ΝαΟΗ 0.029 MgCP 0.104 PVP-12 2.083 Sodium octanoate 3.125 Methyl cellulose 0.104 Hydrophobic Medium Castor oil 52.858 Glyceryl tributyrate 28.466 Ethyl isovaleratc 8.195 Glycerol monooieate 1.779 Lecithin 1.893 Span-40 0.946 B. A formulation for leuprolide: Table 1B presents an example of a composition for an API (Active Pharmaceutical Ingredient) in accordance with one or more embodiments. More specifically, this composition is a leuprolide formulation.

Table IB

Ingredient % w/w

Hvdiuphilic fraction Lxuprolide 0.072 NaOH 0.038 MgCh 0.137 PVP-12 2.740 Sodium octanoate 12.002 Methyl cellulose 0.137 Water 0.605 Hydrophobic Medium Span-40 . 1.21 ' I^eci thin 2.43 Ethyl-iso vale rate 10.52 Glycerol monooleate 2.28 Glyceryl tributyrate 23.74 Castor Oil 44.09 C. A formulation with decreased amount of liv'd ro phobic medium (50% of 5 hydrophobic medium)

Table 1C presents an example of a composition for an API in accordance with one or more embodiments. More specifically, this composition is a formulation for dextran fI‘T)4). The FD4 is HTC-labeled dextran with a MW of 4.4kDa (Sigma, Ff)4) and this is 10 the dextran which was used throughout the Examples unless slated otherwise. This particular formulation contains coconut oil (Sigma) instead of GTB. WO 21)10/(132140 PCr/IB2(IO9/(M17155 39

Table 1C

Ingredient % w/w Hvdrophilic Fraction Dextran 0.939 NaOH 0.001 MgCh 0.235 PVP-12 4.693 Sodium ocianoate 20.662 Methyl cellulose 0.235 Water 1.071 Hydrophobic Medium Span-40 1.04 Lecithin 2.08 Ethyl-iso valerate 9.01 Glycerol-monooleate 1.95 Coconut oil 20.33 Castor oil 37.75 'flic above formula lions are used for a wide variety of therapeutic agents and give good 5 bioavailability to the cargo compound in the animal models described below.

Note that the net amount of therapeutic agent may vary as appropriate in any of the formulations and there may be minor variations in die formulations: for example NaOH is not always used; coconut oil may be used instead of glyceryl tributyrate; MgCb is not always used (e.g. with hCil-I it is not used); all ingredients may be substituted as described 10 above in the specification.

Example 2: Schematic representation of insulin formulation production KIU. I illustrates a method of producing a composition in accordance with one ,j more embodiments. For example, this method may be implemented to make the compositions presented above in Example 1. 15 WO 201(1/032140 1*071820110/007155 40

Example 3: The combination of solid particles containing sodium octanoate and hydrophobic medium is critical for permeation activity.

IdG. 2 presents data relating to serum insulin levels alter rectal administration to 5 rats. Rats were anesthetized and were administered 100 pL of bulk drug formulation ' containing an insulin dose of 328 pg/ral (9 LlJ/rai). Blood samples were collected at 0. 3. 6, 10. 15. 25, 30,40, 60 and 90 minutes post administration and serum was prepared for determination of human insulin by an immunoassay kit with no cross reactivity between rat and human insulin. 10 Data is presented as MEANiSD, n-5. The left panel of FdG. 2 relates to administration of human insulin with sodium octanoate (Na-G8) or solid hydrophilic fraction suspended in water (solid particles in water). The right panel of PIC.». 2 relates to administration of full insulin formulation (solid panicles in hydrophobic medium). Table 2 below1 presents a summary’ of AUC values calculated from die concentration vs, time 15 curves.

Table 2

Test compound AUC(o.u) Na-C8 5753 ±3569 Solid particles in water 4083 ± 2569 insulin in formulation (Solid particles in hydrophobic medium) 280933 ± 78692

Data are MEANiSD 'Die average exposure (expressed by AUC values) to insulin after rectal administration of insult n-SCD was about 50-fold higher than the exposure after 20 administration without a hydrophobic medium. Minimal exposure was detected in ruts administered insulin with sodium octanoate alone or as pan of die solid particles of the hydrophilic fraction (as listed in Example 1) suspended in water. These data demonstrate synergy between solid sodium octanoate and a hydrophobic medium.

Example 4: Intestinal absorption of insulin after Gl administration of insulin to rats 25 FIG. 3 presen is datu relating to serum insulin levels and blood glucose levels after rectal administration of insulin solution and insulin in formulation to rats. Rats were anesthetized and administered 100 μL of test article (insulin in formulation or insulin in WO 2010/0321411 PC'171112009/007155 41 PBS) containing an insulin dose of 328 pg/rat (9 Ι&amp;#971;/rat). Blood samples were collected at 0. 3. 6. 1(). 15. 25, 30. 40. 60 and 90 minutes post administration. Glucose level was immediately determined with a glucometer and serum wus prepared for determination of human insulin by an immunoassay kit with no cross reactivity between rat and human 5 insulin.

Glucose levels are presented as the percentage form basal levels measured before administration (time 0). The data of FIG. 3 is presented as MEANiSD, n=5. 1 rvels of insulin (left panel on FIG. 3) and glucose (right panel of EIG. 3) after rectal administration of human insulin solubilized in PBS (insulin solution) or 10 incorporated in the formulation are presented, insulin levels rose rapidly in rat serum after rectal administration of insulin in formulation. Maximal levels were measured within 6 minutes post administration and a gradual drop delected until reaching basal levels at about 90 min post administration. This sharp and significant rise in insulin was accompanied by a significant drop in glucose levels reaching an average of 2098 of the 15 initial levels already at 30 min post administration. By contrast, rectal administration of insulin in PBS caused only a very slight glucose reduction, which is identical io that observed following treatment with the PBS control alone.

Example 5: Insulin absorption after rectal administration of insulin in formulation to rats 20 FIG, 4 presents data relating to changes in blood glucose and serum insulin concentrations following SC (subcutaneous) administration of insulin solution (at 20 pg/rat) and rectal administration of insulin in formulation (at 328 pg/rat). Blood samples were collected at 0. 3, 6. 10, 15, 25, 30, 40, 60 and 90 minutes post rectal administration and at 0. 15. 30. 45,-60, 90 min, 2, 3, and 4 hours post-SC administration. Glucose was 25 immediately determined with a glucometer and insulin by an immunoassay kit, Glucose levels are presented as the percentage form basal levels measured before administration (time 0), The data of EIG. 4 is presented as MEAN±SD. n=5,

Hie levels of insulin absorption from rat colon after insulin in formulation administration were compared to die levels of insulin absorbed al ter SC administration. 30 Insulin exposure was calculated from the area under the serum concentration versus time curve (AGC) and the activity calculated as the relative bi oavai lability (rBA) according to die following equation: rBA = (rectal AUCrcun) /SC AUC<o-«>) * (SC dose/rectai dose) VV0 201IW«32J4« I*CT/tB21t0y/(H)7l5? 42

Insulin penetration into the bloodstream occurs during a narrow window' of lime, generally within about 10 minutes of rectal insulin in formulation administration. The rise in serum insulin is paralleled by a fall in blood glucose levels. in order to derive information about insulin bioavailability when formulated insulin is 5 presented into the colon. AUCn^was determined for rectal and SC administration and the rBA value of human insulin was 29.4±3.4% with coefficient of variance (CV) - 11.4%.

Rectal administration of various insulin-con taint ng formulations was carried out on hundreds of animals. The assay was further developed and qualified as a bioassay to support platform development and batch release tests with a linear range of 10 -200 10 pg/rat. repeatability of 39% and intermediate precision of 33%. ‘Ihe insulin formulation described herein was tested in five different studies using a total of 25 rats. The rBA was 34.1 ± 12.6% with CV of 28.9%,

Example 6: Insulin absorption after intra-jejuiial administration of insulin in 15 formulation to rats

Tlie absorption target site of the orally administered platform oft he invention is generally die small intestine. To test the activity of insulin formulation in rat intestine, two major obstacles were addressed: 1. Enteric-coated capsules for rats are not available and therefore stomach bypass enabling direct intra-jcjunal administration is needed. 2. 20 Insulin is extensively metaholized by the liver, in humans 50-80% of endogenous insulin, secreted by pancreatic β-cells, is sequestered by the liver and therefore can not be delected in the systemic circulation, Insulin administered via the intestinal route (by way of insulin formulation) mimics the endogenous route of insulin as the intestinal blood flow is drained into the portal vein which leads directly to the liver. Therefore to 25 determine insulin absorbance, blood samples must be drawn from the portal vein (portal circulation, prior to the liver) as well as the jugular vein (systemic circulation, after the liver), A specialized rat model in which three different cannulas are surgically implanted in anesthetized rats was developed: 1. Jejunal cannula - stomach bypass, enables insulin 30 formulation administration, 2. Portal vein cannula - blood sampling prior to the liver, determine insulin that cross die Crl wall into the blood, and 3, Jugular vein cannula - to WO 2010/1)32140 determine the systemic levels of insulin. Using diis model, die bioavailability of insulin in formulation (rBA) was determined. FIG. 5 presents duta from a representative study relating to insulin levels in the portal and systemic circulations after imra-jejunal administration of insulin control and 5 insulin formulation io rats. Rats (8 rats per group) were anesthetized and their jejunum exposed by abdominal surgery. The jejunum containing intestinal loop was placed on gauze and kept moist and fully intact throughout the entire study. A temporary' cannula was inserted into die jejunum and formulated insulin was administered. Blood was collected from both portal and jugular veins at die same time points, with approximately 4 10 time points per rat. The MEANiSD value of each time point was used io create a plasma concentration vs. time curve. AUC was determined and rBA was calculated.

Insulin levels in both the portal and systemic circulation rose dramatically after intra-jejunum administration of insulin in fonnuladon. ’ITits is in contrast to the minimal insulin absorbance detected when insulin control was administered. The window of 15 absorption was short and insulin levels peaked by 6 minutes. This profile is similar to that seen after rectal administration of formulated insulin (see above). Higher insulin levels were detected in the portal compared to the systemic circulation, with rBA of 10,1 % compared to 5.6%. respectively.

Example 7: Additional formulations comprising various cargo compounds 20 Table 3 A details the components of a range of dextran formulations which were prepared as described in the following Examples. The sodium caprate was obtained from bluka/Sigma . the olive oil from Fluka, the ocianoic acid from Sigma and the mineral oil from Acres.

Table 3A . \ | ·: ·υΗΓβ9;·ΤΑ· nnpBB 1 Formulation - ? : >F G H Llngradjeht J wZw) : W/W) j Tar «ο 0.545 0.939 0.565 0.546 j 0.565 0.565 0.565 0.551 WO 2« I«/«3214« 44

NuOH 0.001 0.00! 0.001 0.001 0.0« I 0.001 0.001 0.001 MgCl? 0.136 0.235 0.141 0.156 0.141 0.141 0.141 0.138 Jlvdmpliilk fraction PVP-1? 2,726 4.693 2.823 3.1 17 2.823 2.823 2.823 2.754 Sodium oci annate 12.001 20.662 9.002 9.002 9.( K12 12.125 Sodium cuprate - - 9.002 - MC. 400 0.136 0.235 0.141 0.156 0.141 0.141 0.141 ¢).138 Water 0.622 1.071 0.507 0.159 0.507 0.507 0.507 0.661 Hvdro- phobic medium Spun40 1.21 1.04 1.25 1.38 125 1.25 1.25 1 .ecilhin 2.42 2.08 2.50 2.76 2.50 2.50 2.50 Ethyl i so- valerate 10.46 9.01 10.8? 11.96 10.83 10.83 50.8? 11.23 Glyceryl inonoolenir 2.27 1.95 2.35 2.60 2.35 2.35 2.35 Glyceryl rribiitvrtue 23.62 20.33 24.46 24.29 24.46 24.46 24.46 25,35 Coconut oil - Castor oil 43.86 37.75 45.42 45.07 45,42 47.08 Oetanoii· acid 7.80 Mineral oil - • 45.42 Olive oil | * * 45.42 5 'Table 3B details the components of a range of teriparatide acetate and leuprolide formulations which were prepared as described in (he following Examples. Teriparatide was obtained from Novetide, and leuprolide was obtained from Bambio.

Table 3B ” . " V Λ v.< y \ • - ;>·<>%/·*, 11 S 88388 16Π parat io ^Formulation? &#912;'&#943;Γ&#912;&#912;5·^&#943;>>Λ*&#943;-';&#912; >y,; WVl»vi3&amp; .r.'ti 4? iti ; "&#943;'.Γ'/Κ1 ' ii.:.,. , E sw Cargo 0.118 0.118 0.050 0.050 NuOH - 0.040 o.tw MgCI; 0.137 0.137 0.142 0.15

Hydrophilic fraction PVM2 2.740 2.740 2.838 2.99 Sodium oemnoate 12.001 12.001 9.012 Sodium caprale - 4.48 MC 400 0.137 0.137 0.142 0.15 Water 0.605 0.605 0.489 0.33 Span40 1.214 1.214 1.26 1.32 Lecithin 2.428 . 2.428 2.52 2.65 flvdro- phobic medium BtbyMso* vulerate 10.515 10.515 10.89 11.46 Glyceryl nioiiooleaic 2.283 2.283 2.36 2.49 Glyceryl tnliutymte 23.740 - 24.59 25.87 Coconut oil - 23.740 - Casior oil 44.082 44.082 45.66 48.04 5 &amp;#906; able 3C details the components of hGH formulations which were prepared as described and the following Examples. The hCtll was obtained from PER. Israel (G11P-24). 10

15 Table 3C i. tom : '·:Η ^Formulation'; s1 *$:;w A*. ξ. d &amp;#912;φ&amp;#970;&amp;amp;ΑΟ •v) c ; g ; fek,T,e.- USS sms 11 vdrophil ic fraction Cargo 0.298 0.303 NaOH - - MgCl, • - pvp- 12 2.836 2.738 Sodium ociimoale 9.006 12.007 WO 2011)/032140 46

Sodium cuprate MC 400 0.)42 0.137 Witter 0,492 0.607 Span40 1.257 1.213 Lecithin 2.514 2.427 Hydro- Ethyl-iso- IO.K85 10.508 phobic valerate medium Glvucryl mon uolcaa* 2.363 2.281 Glyceryl tribuivnite 24.575 23.725 Coconut oil Castor oil 45.633 44.054

The production process for all these above formulations is essentially as described in Figure 1 and in Example 11. 5 Example 8: Effect of dose of sodium octanoate incorfiorated in formulation on formulation activity 'Hie effect of increasing the amount of sodium octanoate (Na-C8) in Lite formulation on the activity oflhc formulation was tested using formulations containing 10 dextran (average. MW - 4.4 kDa, FTFC labeled) as cargo compound and different doses ol Na-C8 namely, formulation A in Table 3A (which contains 12% sodium octanoaie by weight) and similar dextrun formulations containing different Na-C8 doses: 9%. 6% and 3% respectively.

To test the activity of these formulations in the jejunum of non-anesthetized rats, a 15 rat model was established in which two different cannulas are surgically implanted in male Sprague-Dowley rats 1- Jejunal cannula to bypass the stomach and enable direct formulation administration to the jejunum. 2- Jugular vein cannula to determine the systematic levels of the administered 20 dextran following jejunal administration. Rats are allowed to recover for 4 days before Lhe study and are deprived of food for 18 hours before the start of the study.

Figure 6 presents data from a study which determines FITC-labeled dextran (4.4 kDa) bioavailabiliLy in non-anesthetized ruts following intra-jejunal administration of 47 formulations containing different amounts of Na-C8 or FfJ'C-labeled dextran solubilized with the Na-C8 in saline solution (control). ’Ihe bioavailability of the different dextran formulations and the control was evaluated by administrating the different formulations directly to the jejunum of non- 5 anesthetized rats and measuring plasma dextran levels at 3. 6, 10. 25. 60 and 90 minutes post administration. Levels of plasma dextran following administration of dextran in formulation or in saline were compared to the levels of plasma dextran idler intravenous administration. Exposure values. AUC (0-90), were determined lor jejunal and intravenous administration and the absolute bioavailability (aBA) was calculated 10 according to the following equation; aBA- (jejunal AUC(0-9O))/ fix’ AUC (0-90))* (iv dose/ jejunal dose). Data are presented as Mean ± SO (n>5 rats per group). 'the results show that increasing the amount of Na-C8 incorporated in the formulation improves ihe bioavailability of the dextran in a dose-responsive manner, reaching almost 15 30% aBA at the 1 2% (w/w) dose. Dextran administered with Na-C8 at similar doses and suspended in a saline solution (i.e. not formulated) showed much lower bioavailability (-6% aBA). Further results dose- response results are shown in Example 26,

Example 9: Effect of the ratio of hydrophilic fraction/hydrophobie medium on 20 formulation activity 'Ihe effect on formulation activity of changing the ratio {weight /weight) between the hydrophilic fraction and the hydrophobic medium was tested using formulations containing dextran (average MW = 4.4 kDa, FITC labeled) as cargo (formulations A and B in fable 3A). Ihe in vivo non-anesthetized rat model described in Example 8 was used 25 in order to compare the activity of the described formulations. fable 4 presents bioavailability data following intra-jejunal administration of formulations comprising a different ratio of hydrophilic fraction to hydrophobic medium.

Table 4

Largo formulation Weight ratio between hydrophilic/ hydrophobic medium Animal model Route of administration N %aB A± SI) i)e strati A 1 /5.2 Rat Non- anestheti zed Jejunal 17 28,0 - 6.ft B 1 12.6 19 24.8 e 25 30 WO 2011)/032140 ^071820(19/007155 48

Formulations A and B were administered directly to ilie jejunum of non-anesthetized rats and plasma dextran levels were measured at 3. 6. 10, 25, 60 and 90 minutes post formulation administration. The levels of dextran absorption from rat jejunum after administration of dextran in formulation were compared to the levels of 5 dextran absorbed after intravenous administration. Exposure values. AUC (0-901, were determined for jejunal and intravenous administration and the absolute bioavailability (aBA) determined according to the following equation: aBA= (jejunal AUC(0-90)l/ (iv AUC (0-90))* (iv dose/ jejunal dose). Dula are presented as Mean ± SD in > 5 rats per group). 10 'Ihe results show that changing the ratio between the hydrophilic fraction and the hydrophobic medium in these formulations with a low % weight of therapeutic agent hud no significant effect on the bioavailability of die cargo which gives a loading flexibility in devising additional formulations.

Example )0: Activity of formulations containing different cargo compounds 15

In order to test die capability of the formulation platform, the activity of formulations containing three different cargo compounds (APIs) was tested in three different animal models: jejunal administration to non-anesthetized rats, rectal administration to anesthetized rats and jejunal administration to non-anestbetized pigs. 20 Table 5 summarizes the results of representative experiments testing the bioavailability of formulations containing different APIs in the three different animal models described above.

Table 5 Λ Pl Formu lation Animal mode) Route <4 Administration | %BA x SI) t Teriparalide 1 km nonanes ihciizcd Jejunal 5 14.0·'* xIO.S El I Pig non- anesthetized Jejunal j j 15.0** 4-tj J Hl Lcuproiidc K Rat nan- anesthetized Jejunal j 4 >0.1* ±7.5 [V hOH I1 Rai anesthetized Rectal j 5 17.9** ±3.9 * Absolute BA (compared to IV) WO 2010/03214» PCI7iB2»»9/»(!7155 49 ** Relative BA (compared to SC) A. Leuprolide absorption after jejunal administration of leuprolide in formulation to rats 5 Table 5-Π1 presents data from a representative study relating to ieuproliile % aBA following TV (intravenous) administration of leuprolide solution (at 75 pg/Kg) and jejunal administration of leuprolide in formulation (at 450 pg/Kg; formulation K. Table 3B) to non-ancsthetized rats, as previously described in Example 8 .

Blood samples were drawn from the jugular vein at 3. 6. 10. 15. 25. 40, 60 and 90 10 minutes post jejunal administration and at 3. 10. 25, 40. 90 min. 2. 3.3 and 5 hours post IV administration, plasma was prepared and leuprolide levels were determined in each sample. Leuprolide levels in systemic circulation rose dramatically after jejunal administration of leuprolide in formulation. Leuprolide blood levels peaked by 3 minutes post administration. The average aBA achieved after jejunal administration of leuprolide 15 in formulation was calculated as described in the above Examples und was 10.1%. in a control experiment, jejunal administration of leuprolide in TBS demonstrated negligible penetration to the bloodstream. A similar leuprolide formulation containing 12%: sodium oeLanoate as described in fable 1 B was prepared; it was tested in the above model and showed bioavailabiJity as 20 follows: rBA (compared to SC) = 2 L1 % ±12.0 (CV=57%). B. Teri para tide absorption after jejunal administration of teri paratide in formulation to rats 'fable 5-1 presents data from a representative study relating io plasma teri paratide

25 concent radon-time profiles following SC administration of teriparaiide solution (at 85 μ g/formulation and jejunal administration of teriparaiide (teriparaiide) in formulation (at 550 pg/Kg : formulation I. Table 3B) to non- anesthetized rats, as previously described in Example 8 . Blood samples were drawn from the jugular vein at 3. 6. 1(1. 25. (SO and 90 minutes post jejunal administration and al 3. 10. 30, 60, 90 min, 2 and 3 hours post SC 30 administration, plasma was prepared and teriparaiide levels were determined in each sample. Teriparaiide levels in systemic circulation rose dramatical])' after jejunal administration of teriparaiide in formulation. Teripararide levels peaked by 3 minutes post-administration. 'Hie average rBA achieved after jejunal administration of teriparaiide WO 2010/03214« in formulation was calculated as described in the above Examples, and was 14.0%. In a control experiment jejunal administration of teriparatide in saline demonstrated no penetration to the bloodstream. C. Teriparatide absorption after jejunal administration of teriparatide in 5 formulation to pigs

Table 5-Π presents data from a representative stud)1 relating to plasma teriparatide con centra tion-time profiles following SC administration of teriparatide solution tai 10.65 pg/Kgl and jejunal administration of teriparatide in formulation (at 100 pg/Kg: formulation I. Table 3B) to non- anesthetized pigs. 10 Λ pig model was established in which two different cannulas were surgically permanently implanted in female domestic pigs: I - jejunal cannula to bypass die stomach and enable direct formulation administration to the jejunum. 2- jugular vein catheterization to determine the systematic levels of the administered 15 cargo following jejunal administration.

Pigs were allowed to recover for 7 days before Lhe experiment and deprived of food 18-20 hours hefore the stall of the experiment

Blood samples were drawn from Lhe jugular vein at 0. 3, 6. 10. 15. 25. 40. 60, 90 minutes, 2. 2.5 and 3 hours post jejunal administration and aL 0. 3. 6. 10. 15. 20. 30, 45. 20 60. 90 min. 2, 2.5. 3 and 4 hours post SC administration, plasma was prepared and teriparatide levels were determined in each sample. Teriparatide levels in systemic circulation rose dramatically after jejunal administration of teriparatide in formulation. Teriparatide levels peaked by 10 minutes post administration. 'Hie average rBA achieved after jejunal administration oF teriparatide in formulation was calculated as described in 25 the above Examples, and was 15.0%. A similar pig experiment was performed using dextran (ED4. formulation Λ in Table 3A) and it was determined that die average bioavailability of dextran was 20% in pigs as compared to IV. D. hGH absorption after rectal administration of hGH in formulation to rats

30 Table 5- IV presents data from a representative study relating to plasma hGH concentration-time profiles following SC administration of hGH solution (at 81 pg/Kg) and rectal administration of hGH in formulation (at 800 pg/Kg; formulation P. Table 3C). io anesthetized rats. WO 201)(/032140 PC171B2009/007155 51

Male Sprague-Dowlev ruts were deprived of food for 18 hours before the start of the experiment. Rats were anesthetized by a solution of ketamine: xylazine. The formulation (lOOpIJ mi) was administered rectally using a 14G venflon. Blood samples were draw'll from the jugular vein at 3, 6, It). 15, 40. 60 and 90 minutes post rectal 5 administration and at 15. 30, 45, 60. 90 min, 2, 3, and 4 hours post SC administration, plasma was prepared and hGH levels were determined in each sample. hGH levels in systemic circulation rose dramatically after rectal administration of hGH in (brmuiaiion. hGH levels peaked by 15 minutes. 'Hie average rBA achieved after rectal administration of hGH in formulation was calculated as described in the above Examples and was 10 17,9%. In a separate experiment hGH was administered to the jejunum and the aBA was lower. In a control experiment, recta] administration of hGH in PBS demonstrated no penetration to the bloodstream.

Ibus the results presented in Table 5 demonstrate that substantial exposure was obtained for all cargo compounds tested in all animal models tested, 15 'Ibe above results demonstrate that the formulations described herein enable delivery of a wide range of different macromolecules through the intestinal epithelium in different animal models.

Example Π: Detailed production process of a formulation of teriparatide 20

Production of the hydrophilic fraction: To 200 mL water die following ingredients were slowly added one by one (with 2-3 minutes mixing between each ingredient): i 72 mg of teriparatide, 200 mg of MgCb, 4.0 g of PVP-12, 17.52 g of sodium octanoate and 10,0 g of 2% MC-400 aqueous solution, prepared as follows: 1 g of MC-400 powder was 25 added to 50 mL w'ater at 60*2 *C while mixing. After 5 min of mixing, the beaker was transferred to ice until a clear solution was obtained.

After addition of the MC-400 solution, the solution was mixed for another 5 min and then lyophilized for about 24 h. This procedure produced about 22g of hydrophilic fraction, 30 Production of the hydrophobic medium: 2 g of Span 40, 4 g of lecithin and 3.8 g of t&amp;#938;ΜΟ were dissolved in 17.3 g of ethyl isovalerale while mixing. To this solution were added 39.1 g of GTB and 72,6 g of castor oil. This procedure produced about 136-138 g of hydrophohic medium. WO 2010/03214(1 52

Prod ucl ion of the bulk drug product: Mixing of the hydrophilic fraction and the hydrophobic medium was performed at 20±2 UC. 15.7 g of the hydrophilic fraction was slowly added during mixing io 84.3 g of hydrophobic medium at 600+50 RJPM. After addition of ail the hydrophilic fraction, the 5 mixing speed was increased to 2000+200 RPM for 2-10 min followed by 4-8 cycles of 15 min mixing at 600±50 RPM and 2 min mixing at2000+200 RPM.

Degassing by vacuum was then applied as follows: 5 min at 600 nt Bar. 5 min at 500 mBar and 30 - 120 min at 400 mBar. Ihe resulting suspension was poured into a 100 ml. dark boule and stored at 2-8°C. This is Lhe teriparatide formulation designated “Γ 10 described in Table 3B.

All other formulations described herein were produced by this method, varying ingredients and quantities according to the details given in the relevant Tables (see e.g. Example 29). A diagram of this method (with insulin as cargo) is shown in Figure 1. Example 12: Effect of the oil incorporated in the formulation on formulation activity 15 The effect of the type of oil incorporated in the formulation (in the hydrophobic medium) on formulation activity was tested. Formulations containing dextran (average MW = 4.4 kDa. FFPC labeled) us cargo compound and different types of oils in the hydrophobic medium (formulations E, F and Ct in 'fable 3A).were tested in rats.

To lest the activity of these formulations in die jejunum of non-anesthetized nits, a 20 rat model was established in which two different cannulas are surgically implanted in male Sprague-Dow] ey ruts: 1- Jejunal cannula to bypass the stomach and enable direct formulation administration to the jejunum. 2- Jugular vein cannula to determine (lie systematic levels of the administered 25 dextran following jejunal administration.

Rats are allowed to recover for 4 days before the study and are deprived of food for 18 hours before the start of the study.

Table 6 presents data from a study in non-unesthetized rats following intra-jejunal administration of formulations containing different oils in the hydrophobic medium.

Table 6

Cargo Formulation Oil N % aBA ±SD Dextrun E Castor oil +GTB 14 19.8 ± 5.5 P Mineral oil + GTB 5 12.2 ±5,0 Cl Olive oil + GTB 5 ' 12.0 ±9.9

Formulations containing different oils were administered directly to the jejunum 5 of non-anestheiized rats and plasma dextran levels were measured at 3. ft. 10. 25. ft() and 90 minutes post formulation administration. The levels of dextran absorption from rat jejunum after administration of dextran in formulation were compared to die levels of dextran absorbed after intravenous administration. Exposure values. AUC (0-90). were determined for jejunal and intravenous administration and the absolute Bioa variability 10 (aBAj was determined according to the following equation: aBA= (jejunal AUC(0-90))/ (iv AUC (0-90))* (iv dose/ jejunal dose). Data arc presented as Mean ± «SD (n > 5 rats per group).

Similar bi «availability was achieved when dextran was incorporated into formulations containing castor oil or coconut oil. Good hioavailahility was also obtained 15 in rat jejunum when teriparatide was used as cargo compound using formulations I and .1: these formulations contain castor oil and GTB, and castor oil and coconut oil. respectively.

The results showed that formulations containing different kinds of oils in their hydrophobic medium are active, enabling penetration of the cargo (dextran, teriparatide) 20 earned by the formulation. Thus the data demonstrated that all tested oils enable bioa vailahility of the cargo carried by die formulation. Castor oil and coconut oil might be superior to the other tested oils.

Example 13: Preparation of a formulation using granulation instead of lyophilization 25 Production of the hydrophilic fraction: To a plastic bag. the following ingredients were added: 1.00 g of PVP-30. 6.70 g of sodium octanoate and 13.00 g of lactose WO 2011)/()32141) monohydralc its binder. After 5 nnn of mixing, all of Lhe powder was transferred into a mortar and pestle. Λ dextran ED4 aqueous solution was prepared as followed: 0.42 g dextran was dissolved in 1.2g of WFL AU of die dextran solution was dien added slowly to the powder 5 while using a low shear agitation in a morutr &amp;amp; pestle: the agitation took around 45 min, Hie mixture was then transferred into a lyophilization nay and was oven-dried for about 20 h at 50”C. This procedure produced about 20g of hydrophilic fraction, which was a fine granulate.

Production of the hydrophobic medium: 2 g of Span 40. 4 g of lecithin and 3.8 g of 10 GMO were dissolved in 17.3 g of ethyl isovaleraie while mixing. To this solution were added 39.1 g of GTB and 72.6 g of castor oil. This procedure produced ahout 136-138 g of hydrophobic medium.

Production of the bulk drug product: Mixing of the hydrophilic fraction and the hydrophobic medium was performed at 20±2 UC. 15 19.00 g (29.58% of the final BDP) of die hydrophilic fraction was slowly added during mixing to 45.23 g (70.42% of the final BDP) of hydrophobic medium ai 600±50 RPM. Aher addition of all die hydrophilic fraction, die mixing speed was increased to 2000±200 RPM for 2-10 min followed by 4-8 cycles of 15 min mixing at 600*50 RPM and 2 nnn mixing at 2000±200 RPM. 20 Degassing by vacuum was then applied as follows: 5 min at 600 mBar. 5 min at 500 mBar and 30 - 120 min at 400 tnBax. 'lhe resulting suspension was poured into a I (1(1 mb dark bottle and stored at 2-8°C.

Rat study: 'lhe above suspension was administered racially to rats as described above in the Examples and the results were as follows: 35 %BA, 12.9 %SD. Another batch of 25 suspension prepared by granulation us described above was prepared and was administered to die jejunum of rats as described above in the Examples, and the results were as follows: 21.8 %BA, 4.0 %SD.A range of formulations are prepared in a similar manner using granulation and incorporating a selection of therapeutic agents and varying the amount of sodium octanoate. 30 Example 14: Selection of capsules

In vitro experiments were carried out using separately three types of solutions: the hydrophobic medium as described in die above Examples, ethyl isovaleraie alone, and ethyl isovaleraie containing 5% of each of the following surfactants: lecithin, span 40 and WO 2010/032140 t’CT/l &amp;#970;&amp;#943;2009/007155 55 glyceryl mono-oleate. 3 types of unsealed capsules, gelatin, starch and HPMC. were each filled with each of these solutions, lhe filled capsules were then maintained in vitro for 29 days ai 22±2"C. 30-50% relative humidity. Gelatin and HPMC. capsules gave the best results, namely no deformation of die capsule. 5 Similar experiments were carried out using the same three solutions, and gelatin and HPMC capsules. The capsules were filled with die solutions, sealed (bonded) and then were maintained for 8 days at 22±2°C, 30-50% relative humidity. Both types of capsules showed stability to the solutions tested i.e. there was no leakage and iki deformation of the capsules.

Hi

Example 15: Effect of varying the cation in the medium chain fattv acid salt

Formulations were prepared with dextran (FD4) similar io Formulation A of Tabk 3A except that 12%; sodium ocianoate (0.722M) was replaced by an equal molarity of lithium ocianoate or potassium ocianoate or arginine ocianoate (the last as a model for an 15 ammonium salt). These formulations are shown below in Table 7A.

Table 7A

Formulation: cargo =dextran>? - MngrudientW£&amp; LWiiSWSJSSiif ^K^iCiajipai^^ Xs·? ^Afi^dctanoate··;??;'; ·Λ&#939;<Γ V ·4χ :«W SSSii'S ν •λ7·'?,;·'\·;·····;·,'.<·(&#943; i(%?w7w);: ν,?/·'^<· &#943;γ * , i 1 ydrttpiiilie t ract km API 0.545 0.546 0.546 MgCI2 0.134 0.136 0.124 PVP-12 2.673 2.722 2.475 Potassium ocianoate 13.617 0.00 0.00 Lithium ocianoate 0.00 10.826 0.00 Arginine ocianoate 0.00 0.00 22.989 MC 400 0.134 0.136 0.124 Water 0.684 0.627 0.919 Span40 1.185 1.206 1.097 lecithin 2,369 2.412 2.193 WO 2010/032140 PCI7IB21HW1M17155 56 I lydiuphubie medium Ethyl iso valerate 1(1.26 10.45 9.50 Glyceryl monooleate 2.227 2.268 2.062 Glyceryl tri butyrate 23.16 23.58 21.44 Castor oil 43.01 43.79 39.82 'lliese formulations were each tested in the rat jejunal model described in Example 8. TTic results were obtained and bioavailability was calculated. 'Ihe results are shown 5 below in Table 7R.

Table 7B

Medium chain fatty acid salt in formulation tested N %BA± SI) Sodium octanoate (Formulation Λ) 18 22.2 ± 10.8 Lithium octanoate 11 8.4 ± 3.8 Potassium octanoate 10 7.9 ± 6.4 Arginine octanoate 12 17.5 ± 7.4

Ihe formulation A used in the above experiment was a different batch to that used 10 in Example 8. and so die BA results given here for formulation Λ differ slightly from those recited in Table 4.

The above results show that when 12% sodium octanoate was replaced in the formulation by an equivalent molarity of lithium octanoate or potassium octanoate . the formulation still had bioavailability but at a lower level. The arginine octanoate 15 formulation had similar activity to the 12% sodium octanoate formulation. WO 20111/032140 PCT/IB 2009/007155 57

Example 16': Effect of addition of medium chain alcohols (geraniol and .oetanol) to the hydrophobic medium.

Formulations containing geraniol (BASF) and oetanol (Spectruni/MP) were 5 prepared as described above, using die ingredients shown below in Table 8. The sodium dodecanoate was obtained from Speelruni/Acros).

Formulation 0- low % medium chain fatty acid salt; A dextran (PD41 formulation was prepared essentially as described in Example 11. containing a total of 2.9% medium chain fatly acid salt - (sodium octanoate 1.042% 4- sodium dodecanoate 1.869%) -and 10 also containing geraniol and oetanol in the hydrophobic medium, all as shown in fable 8 below.

Formulation R- over 10% medium chain fatty acid salt; A dextran formulation was prepared essentially as described for Formulation A except that geraniol and oetanol were added to the hydrophobic medium, all as shown in Table 8. 15 Table 8 W^bext'rah>^'?; IWfe 7Dextran'·' ’ "' J stest KOSW -/λ?·? f 1 Hydrophilic fraction API 0.545 0.456 NaOH 0.029 0.000 MgCb 0.104 0.114 PVP-12 2.083 2.282 Sodium octanoate 1.042 10.046 Sodium dodecanoate 1.869 - MC400 0.104 0.114 Water 0,231 0?521 Hydrophobic medium Geraniol 9,148 8.39 Oetanol 8.627 7.92 Spnn40 1.041 0.96 Lecithin 2.081 1.91 Ethyl isovalerate 9.012 8.27 Glyceryl monooleate 1.956 1.80 Glyceryl tributyrate 21.825 20.03 Castor oil 40.532 37.20 WO 2010/032140 lJCI7Ui200‘J/007155 58

Formulation Q (low % MCFA suit) was tested io the intra-jejunal rat model described above and die bioavadahiJitv was calculated; aBA= 4.4%). Sl.>— 3.8 (n=l 2). Formulation R (over 10% MCFA salt) was tested in the intra-jejunal rat model described above und the bioavailability was calculated: aBA= 22.7%?. SE)= 1.6 (n=6). 'liie BA of 5 these formulations do not differ significantly from similar Fonn ul a lions, described in the above Examples, which do not contain geraniol.

Example 17: Formulations for gentamicin and for RNA

Formulations were prepared for gentamicin and for RNA essentially as described in Example 1 i. with the ingredients of the bulk drug product as shown below in 'fable 9. 10 'Die gentamicin was obtained from Applichem and the RNA was polyinosinic-polvcytidyiic acid sodium salt (Sigma).

Table 9A ^Formulatipn^A^' iii^lngredtent^^e :B N ft ft ’' ' ',,&#943;; Wwiw/W T5·&#943;&#970;- 5&#970;>}>' -aL >»(Hw/wJ · · tlydruphilic fraction AP) 6.000 0.100 NaOH 0.670 MgCI, 0.127 0.137 PVP-12 2.545 2,741 Sodium octanoate 12.026 12.001 MC400 0.127 0.137 Water 0.860 0.605 Hydrophobic medium Span40 1.119 1.214 Lecithin 2.238 2.429 Ethyl isovalerate 9.69 10.52 Glyceryl monooleate 2.103 2.283 Glyceryl tri butyrate 21.88 23.74 Castor oil 40.62 44.09 15 The gentamicin formulation was tested in the rat jejunal mtxlel described above and in the rat rectal model described above (e.g. Examples 4 and 5), The gentamicin was assayed using an immunoassay (ELISA). 'Die results are shown in 'fable 9B below; % ΒΛ is calculated compared comparing to IV administration, 'Hie formulations wen* shown to provide bioavailabi lily to die gentamicin. WO 201 »/«3214» PCr/l82»«y/IMI7t55 59

Table 9B

Cargo Formulation ROA N % BA ± SD Gentamicin As Table 9A jejunal 6 12.9 + 4.5 As Table· 9A rectal 5 50.1 + 5.8

Similarly, die RNA formulation of Table 9A is tested in the rat jejunal model and in the rat rectal model described above. The RNA is assayed and the formulation is 5 expected to provide bioavailability lo the RNA,

Example 18: Effect on formulation activity of the surfactants in the hydrophobic med ill in ‘Ihe effect on formuladon activity of withdrawing surfactants from the hydrophobic medium was tested using formulations containing dextran (average MW = 10 4.4 kDa. WTC labeled) as cargo (formulations A and H in Table 3A).

Table 10 presents data from a study in non-anesthetized rats following intra-jcjunal administration of formulations with or without surfactants (e.g. Span40. lecithin, glyceryl monooleatc) in the hydrophobic medium.

Table 10

Cargo Formulation Surfactants in hydrophobic medium N % aBA ± SI) Dextran A 4- 17 28.0 ± 6.8 H - 4 ll.l +8.2

Formulations with or widiout surfactants in die hydrophobic medium were administered directly' to the jejunum of non-anesthetized rats and plasma dextran levels were measured at 3. 6. 10. 25. 60 and 90 minutes post formulation administration, The levels of dextran absorption from rat jejunum after administration of dextran in 20 formulation were compared to the levels of dextran absorbed after intra venous administration.

Exposure values. AUC (0-90), were determined for jejunal and intravenous administration and the absolute bioavailability (aBA) was determined according io the following cquaiion;aBA= (jejunal AUC(0-9()))/ (iv AUC (0-90)1* Civ dose/jejunal dose). 25 Data are presented as Mean aBA ± SD. 60 I tower bioavailabiliiy was achieved when dextran was incorporated into a formulation not containing surfactants in the hydrophobic medium (formulation II) as compared to a formulation containing surfactants in the hydrophobic medium (formulation A), 'Hie results demonstrate that withdrawing surfactants from the 5 hydrophobic medium adversely affects formulation activity.

Example 19: Effect on formulation activity of withdrawing medium chain fatty acids from the hydrophilic fraction.

The effect on formulation activity of withdrawing medium chain fatty acids (MGFA) from the hydrophilic fraction was tested using formulations containing dextran 1 () (average MW -4.4 kDa, FTI’C labeled) as cargo .

Table 11 presents data from a study in non-anesthetized rats following intra-jejunal administration of formulations with or without sodium octanoate in the hydrophilic fraction (formulations A and D in Table 3A, respectively),

Table! 1 | Cargo Formulation MCFA in hydrophilic fraction N % aBA ± SD | Dextran A + 17 28,0 ± 6.8 D - 5 0.6 x 1.0

The formulations described above were administered directly to the jejunum of non-unesthetized rats and plasma dextran levels were measured at 3. 6, 1(1, 25. 60 and 90 minutes post formulation administration, 'Hie levels of dextran absorption from rat jejunum after administration of dextran in formulation were compared to the levels of 20 dextran absorbed after intravenous administration. Exposure values. AUC (0-90). were determined for jejunal and intravenous administration and the absolute bioavailability (aBA) was determined according to the following equation: aBA= (jejunal AUC(0-90))/ (iv AUC (0-90))* (IV dose/ jejunal dose). Data are presented as Mean aBA ± SD. 25 Negligible penetration of dextran was achieved when dextran was incorporated into a formulation lacking medium chain faLty acids in the hydrophilic fraction (formulation D, % aBA=0.6 ± 1.0) as compared to a formulation containing sodium octanoate at 12% w/w in the hydrophilic fraction (formulation A, % aBA= 28,0 ± 6.8 ), WO 20111/03214(1 Ρθ7ΙΒ2009/0(Π155 61

The results demonstrate dial a formulation without medium chain fatty acids in the hydrophilic fraction is not active. A similar experiment was performed using octreotide its cargo in the improved formulation (see below). The rBA was 0.11% (CV= 158%) 5 Example 20: Effect on formulation activity of simplifying the formulation

The effect on formulation activity of simplifying the formulation was icsted using formulations containing dextran (average MW = 4.4 kl)a. FO'C laheled) or octreotide (Novctide) as cargo. The basic formulation described in die above Examples (e.g. formulations designated A. I and P) was simplified by not adding MgCB. and MC 400 to 10 the hydrophilic fraction and by not adding span40. lecithin and ediyl iso-valerate to the hydrophobic medium. There is a concomitant increase in the amounts of glyceryl tnonooleale (surfactant) and glyceryl tributyrate added to die hydrophobic medium. Such formulations are shown in Table 12A below. These simplified formulations show no precipitation visually although the particles are visible microscopically i.e. they are stable 15 suspensions.

Table 12A - fee miSimpUfted^c v· kwiiSinipiifietd ilBSsei -7 T1? »(»w?wiT-.;F 'rv· - ;. Hydrophilic fraction API 0.545 0.058 NaOH 0.001 0.000 MgCli 0.000 0.000 PVP-12 2.735 2.750 Sodium octanoate 12.000 12.019 MC 400 0.000 0.000 Water 0.S11 0.593 Hydrophobic medium Span40 0.00' 0.000 Lecithin 0.00 0.000 Ethyl isovalerate 0.00 0.000 Glyceryl monooleate 5.91 5.947 Glyceryl tributyrate 34.19 34.385 Castor oil 44.00 44.248

The production process for these above simplified formulations is essentially as 20 described in Figure 1 and in Example 11 for the basic formulations. 'Hie basic octreoude formulation is shown in Table 12B below. WO 2(110/113214(1 62

5 Table 12B W£»rK<t» iVOctrcotideA is; iEormijlaUuDL J ‘;M ' ? ^ineivdlcnt-A, .Τ·:,&#912;Τ?Λ~·;;^&#943;&#912;&#912;Λ&#912;&amp; v.” S/ffiiwAF)’·''· "·> Hydrophilic fraciion (HFP) Cargo 0.058 NaOil 0.000 MgCh 0.137 PVP- 12 2.742 Sodium Octanoate 12,003 MC 400 0.137 Wamr 0.603 Hydro phobic fraction (LFP) Span40 1.215 1-ccithin 2.430 Hihyl-Iso- valcrate 10.522 Glyceryl Monooieate 2.284 Glyceryl Tributyrate 23.756 Castor oil 44.113

Table 13 presents data from a study in non-anesthetized rats following intra-jejunaJ administration of two different dextran formulations - formulation A of Table 3 A 1(1 and the simplified formulation shown in Table 12A.

Table 13

Cargo Formulation N ADC (0-60 min l/dose/kg b.w.iSD Dextran A(hasic) 28 67062 ± 27368 Simplified 12 63897 + 24210 WO 2010/032140 I*CT/IB2OU9/&lt;M7155 63

The above results show that similar AUC values were achieved when dextran was incorporated into a formulation containing Lhe basic formulation (formulation A) as compared to a simplified formulation. 'fable 14 below presents data Γγ&amp;#972;ιιι a study in non-anestheuzed rats following 5 intra-jejuna] administration of two different octreotide formulations - the basic formulation shown in 'fable 12B and the simplified formulation shown in Table 12A. 'Phe levels of octreotide absorption from rat jejunum after administration of octreotide in basic formulation and simplified formulation were obtained. Exposure values. AUC (0-25), were detemlined. 10 Table 14

Cargo Formulation N AUC (0-25 minl/dose/kg b.w. ± SD Octreotide Basic 13 2.8 ± 1.4 Simplified 13 2.3 ±0.8 'ihe above results in Table 14 show that die AUC values were slightly less when octreotide was incorporated into a simplified formulation as compared to the full formulation. 15 Example 21: Effect on formulation activity of replacing castor oil by octanoic acid.

The effect on formulation activity of replacing castor oil (and giyceryi tributyrate and ethyl iso-valerate) by octanoic acid ( Aldritch) was tested using a formulation containing dextran as cargo. This was done to maintain the C8 motif in die formulation i.e. it was considered it might be advantageous to have C8 acid in Lhe hydrophobic 2(1 medium in addition to the C8 salt in the hydrophilic fraction .

The effect of adding ricinolcic acid (Spectrum) was also tested by making a dextran formulation containing octanoic acid/ ricinolcic acid. Ricinolcic acid was chosen since the main triglyceride component in castor oil is formed from ricinolcic acid.

Ihrec formulations of dextran were prepared as shown in Table I5A below. The basic 25 dextran formulation was prepared essentially as described in the above Examples. The dextran octanoic formulation was prepared essentially as described in the above Examples hut wherein castor oil. glyceryl tributyrate and ethyl iso-valerate were replaced by octanoic acid. This formulation was found to be a solution by visual analysis but true solubility analysis was not performed. Il seems that the octanoic acid at high 30 concentration (about 78% of this formulation) dissolves the solid hydrophilic fraction.

FdYIB2IH>«l/007l55 64 with the PVP and sodium octanoate being soluble in oetanoic acid at high concentration, dextran ricinoleic/octanoic acid formulation was prepared essentially as described in the above Examples but wherein castor oil. glyceryl iributyratc and ethyl iso-valerate were replaced by a mixture of oetanoic acid and ricinoleic acid. This formulation was a 5 suspension as is usuu! for most of the formulations of this invention.

Table ISA .1 .-. ft . ,,. Formulation,'APtivri: r &#974;·^!πβ™?!.&#912;βΜ&amp;^ Dextran»·; M ^^PeirtranjHt. ,* S?Gctiihoic'a£id -t > iSspWiK;^? 5»?; wtr; t &#971; I. ;&#943; S •«.“''i· ^Dextran ; > , : Ricinoleic/Octanoic '· c. ' ?·,>.>/ acid.·- · ; SSBS iEv·'.'.((%«/«) Hydrophilic fraction API 0.545 0.545 0.545 NaOH 0.001 0.001 0.001 MgClj 0.136 0.136 0.136 PVP-12 2.726 2.726 2.726 Sodium octanoate 12.001 12.002 12.002 MC 400 0.136 0.136 0.136 Water 0.622 0.622 0.622 Hydrophobic medium Span40 1.208 1.207 '1.207 lecithin 2.416 2.414 2.414 Ethyl isovaterate 10.46 0.00 0.00 Glyceryl monooleate 2.271 2.272 2.272 Glyceryl trlbutyrate 23.62 0.00 0.00 Castor oil 43.86 0.00 0.00 Oetanoic acid 0.000 77.94 23.38 Ricinoleic acid 0.000 0,00 46.76 Ethyl Octanoate 0.000 0.00 7.80 &#906;'he formulations described above in 'fable 15A were administered directly to the 10 jejunum of non-anesthetized rats, and plasma dextran levels were measured post formulation administration. Exposure values. AUC, were determined lor the different formulations. 'Phose results are shown below in Table 15B. 15 WO 2(ΠΙΜ»32Ι4« 65

Table 1SB

Cargo Formulation N AUC (6*60)/&lt;lose/kg h.w. ± SD Dextran Basic 12 72385 ±37827 Octanoic acid 11 180824 ± 32778 Ricinoleic/ Octanoic acid 11 113204 ±33057 10 ' Ihe results shown above in Table 15B demonstrate that Lite absorption of dextran was much improved (over two-fold) in the formulation containing octanoic acid. Additionally, the shape of the graph was changed showing slower but longer release, This may be advantageous since this allows the API to be longer-acting in the body1, The dextran ricinoleic/octanoic results showed less activity than die octanoic acid formulation, 15 hut was still improved over the basic formulation.

Since the octanoic acid and ricinoleic acid/octanoic acid formulations showed high activity, similar formulations were prepared with exenatide as cargo. Three formulations of exenatide were produced as shown in fable 16Λ below. Ihe basic exenatide formulation was prepared essentially as described in the above Examples. The 20 exenaiidu/nctanoic formulation was prepared essentially as described in the above

Examples but wherein castor oil, glyceryl tributyraie and ethyl iso-valerate were replaced by octanoic acid. This formulation containing about 78% octanoic acid was found to be a solution by visual analysts, as was the similar dextran formulation above. 'Ihe exenatide rieinoleic/oetanoic acid formulation was prepared essentially as described in the above 25 Examples but wherein castor oil, glyceryd tributyrate and ethyl iso-valerate were replaced by a mixture of octanoic acid and ricinoleic acid. 30 wo 2010/03214(1 PCT/IB 2009/00715? 66

Table 16A.- ti ,- -. f .i.v sI^Fdhn&amp;iatioivARIOi Si 1MBOT ^pjctanoKaacWijs sxiS :·&#943;.&#973; &#974;>Λ&#943;Γ?&#943;Ν t < ,< if VRicinoieic/Octanoic. Hydrophriic fraction API 0.055 0.055 0.055 NaOH 0.000 0.000 0.000 MgCi; 0.157 0.137 0.137 PVP-12 - 2.742 2.742 2.742 Sodium octanoate 12.003 12.003 12.003 MC 400 0.137 0.137 0.137 Water 0.603 0.603 0.603 Hydrophobic medium 5pan40 1.213 1.214 1.214 Lecithin 2.434 2.429 2.429 Ethyl isovalerate 10.522 0.000 0.000 Glyceryl monooleate 2.283 2.285 2.285 Glyceryl tributyrate 23.759 0.000 0.000 Castor oil 44.112 0.000 0.000 Octanoic acid 0.000 78.395 47.035 Ricinoleic acid 0.000 0.000 23.518 Ethyl Octanoate 0.000 0.000 7.B42 'Hie formulations described above in Table 16A were administered directly to the 5 iejunum of non-anesthetized rats, and plasma exenatide levels were measured post formulation administration. Exposure values, AUC, were determined for the different formulations. These results are shown below in Table 16B,

Table 16B

Cargo Formulation N AUC &lt;0-9«) ± SI) % ΒΛ * SI) Exenatide Basic 10 1961 ±1791 K.8±8.2 Octanoic acid 11 612*350 [Λ&amp;#943;/C (0-180) ±SU! 3.1 ± 1.8 Ricinoleic/ Octanoic acid 9 476*321 2.2 ± 1.5

KJ 'The results shown above in Tuble 16B demonstrate that the exenatide formulation containing octanoic acid showed bioavailabiliiy, but the absorption of exenatide was decreased compared to the basic formulation. The shape of the graph was changed wo 201(1/032141! showing slower but longer release as in die case of the dextran octanoic acid f ormulation above: this prolonged PK profile may be advantageous. Note dial in the case of die octanoic acid formulation. AUC 0-180 min was used for BA calculations due to (he prolonged PK profde. The exenatide ricinolcic/ocianoic acid formulation had even lower 5 bioavailability than the octanoic acid fonnuladon.

Example 22: Dose response for octanoic acid. A. Octreotide formulations: The effect on formulation activity of varying the amount of octanoic acid was tested using font tul alio ns containing octreotide as cargo, &amp;#943; our formulations of octreotide were prepared using 0%. 5%. 10% or 15% ocianoic acid IQ as shown in 'fable 17 below. Tire formulations are basic octreotide formulations prepared essentially as described above wherein the amount of octanoic acid varies as described and the amount of other ingredients in the hydrophobic medium, (ethyl isovalerate and glyceryl tributyrate ) was concomitantly reduced. (In these formulations the hydrophilic fraction was simplified to omit MgC^and MC400.) 15 Table 17 T FoTmulotlbnj'API ^^Inaredlent^^i MOcireotiaetSfeiC jQ%Octandlc?|f59 ictreotide.i fcOctanoiCy iUiOctrebtfae ‘.U l0%Octan0ic ·&lt; ? ^Octreotide" /ISKOctaholc./ i»wZw$i API 0.05 S [ 0.057 0.057 0.057 Hydrophilic fraction PVP-12 2.750 J 2.750 2.750 2.750 Sodium octanoate 12.019 i 12.034 12.034 12.034 Water 0.593 | 0.S94 0.594 0.594 5pan40 1.217 [ 1.219 1.219 1.219 Lecithin 2.441 [ 2.437 2.437 2.437 Ethyl isovalerate 10.554 | 0 0 0 Hydrophobl Octanoic acid 0 1 5.053 10.553 15.021 c medium Glyceryl monooleate 2.290 j 2.291 2.291 2.291 Glyceryl tributyrate 23.832 | 29.325 23.825 19.357 Castor oil 44.246 j 44.241 44.241 44.241 K. Exenatide formulations: 'Hie effect on fonnuladon activity of varying (he amount of octanoic acid was tested using fonnulations containing exenatide as cargo.

Five formulations of exenatide were prepared using 0%. 10%. 15%, 20% or 35% octanoic 20 acid as shown in Table 18 below. The fonnulations are basic exenatide formulations prepared essentially as described above wherein the amount of octanoic acid varies as described and the amount of other ingredients in the hydrophobic medium (ethyl isovalerate and glyceryl tributyrate ) was concomitantly reduced. WO 2()11)/(132140 &amp;#908;’/ΙΒ2(Κ)9/Ο()7155 68

Table 18 . rAG^^FormUtatibn^Ahl^ : : X G '4 SExenatides PSP Gctan ο&#970; c <j£ ^Exenatide·; tea ^Exenatide'· rl5% &#940;&#943;&#943;&#912;-i^OranoicT ^^teriatWe^ .&#912;·Α&#912;;20%.>Α<;:&#943; '^Octanoic’) Exenatide' U-^35%% Octanoic ;; OwZiW q%w/w) 1 API 0.055 0.055 0.055 0.055 0.055 1 MgCI, 0,137 0.137 0.137 0,137 0.137 Hydruptillic j pyp_ 2.742 2.742 2.742 2.742 2.742 J Sodium octanoate 12.003 12.003 12.003 12.003 12.003 | MC 400 0.137 0.137 0.137 0.137 0.137 [ Water 0.603 ' 0.603 0.603 0.603 0.603 t Span40 1.213 1.213 1.213 1.213 1.213 f lecithin 2.434 2.434 2.434 2.434 2.434 j Ethyl isovalerate 10.S22 0 0 0 0 ,. ,. Octanoic acid 0 10.522 15.081 20.085 34.282 c medium 1 Glyceryl monooleate 2.283 2.283 2.283 2.283 2.283 j Glyceryl tributyrate 23.759 23.759 19.201 14.197 0.000 ( Castor oil 44.112 44.112 44.112 44.112 44.112

The formulations described above in Tables 17 and 18 above were administered 5 directly to the jejunum of non-anesthetized rats, and plasma octreotide or exenatide levels were measured post formulation administration. Exposure values, AUC , were determined lor the different formulations. These results are shown below in Table 19. 10 »2009/0117155 69

Table 19

Cargo Formulation N AUC (0-60)/dose/kg h.w, ± SI) Octreotide Basic 14 2.8 ± 1.0 Basic 5 % Octanoic acid 12 2.7 ± 1.2 Basic) 10 % Octanoic acid 12 3.2 ± 1.2 Basie 15 % Octanoic acid 12 4.5 ± 2.3 Exenatide Basic 10 3.9 ±3.8 Basic, 10 % Octanoic acid 15 4.6 ± 2.8 Basic , 15 % Octanoic acid 6 3.0 ± 1.8 Basic . 20 % Octanoic acid 5 2.2 ± 0.5 Basic , 35 % Octanoic acid 6 1.9 ± 0.7 ‘Ihe results shown above in Tublc 19 demonstrate that the octreotide formulation shows increased activity compared io the basic formulation as the amount of octanoic acid is increased to 15% (the maximum amount tested). Additionally, the results shown 10 above in Table 19 demonstrate dial die exenatide formulation shows increased activity compared to the basic formulation as the amount of octanoic acid is increased to 15% and the activity decreases at higher levels of octanoic acid.

Example 23: Effect of different medium chain fatty acid salts. A. Sodium sehacate (disodium sail of decanedioic acid): Ihe effect on 15 formulation activity of replacing sodium octanoate by sodium sehacate (disodium CIO salt I in a dextran formulation was tested, Ihe sodium sehacate was prepared in situ from scbacic acid (Aldrich) and sodium hydroxide. Ihe formulation produced is described in Table 20 below. The formulation was prepared essentially as described above but 12% sodium uctanoate was replaced by sodium sehacate, al llie same molar concentration us 20 sodium octanoate i.e. an equimolar amount of sodium sehacate was used (viz., 0.72M). 0)9/007155 70

Table 20 «a»» WbfeiM LLFormulation/API ^^ngre^teit^^^s ifily'vBbejitfanr" : gy.-'iNa'iSebacaie , Hydrophilic fraction API 0.545 NaOH 0,000 MgClj 0.129 PVP-12 2.589 Sodium Sebacate 18.190 MC 400 0.129 Water 0.783 Hydrophobic medium Span40 1.147 Lecithin 2.295 Ethyl Isovalerate 9.94 Glyceryl monooleate 2.157 Gtyceryl tributyrate 22.44 Castor oil 41.66

The Ibmiulation described above in Table 20 was administered directly to the jejunum of non-anesthetized rats, and plasma dextran levels were measured post formulation administration. Exposure value. AUC. was determined for the formulation 10 and this is compared with a similar formulation prepared with sodium octanoate. JTiese results are shown below in Table 21.

Table 21

Cargo Formulation N AUC (0-60)/dose/kg b.w. ± SI) Dextran Willi Na-octiinoate 1.2 72385 ±37827 With Na-Sebacate 9 18691 ±11887

The results shown in Table 21 demonstrate that the dextran formulation containing 15 sodium sebacale showed activity, but the absorption of dextran was decneitsed compared to the. formulation containing an equimolar amount of sodium octanoate. WO 2011)/()32140 B. Mono-sodium subenne or di-sodium suberaie

Octreotide-containing formulations were prepared wherein 12% sodium ocmnoaie was replaced by an equimolar amount (0.72M) of mono-sodium subenne or of di-sodium 5 suberaie, which are C'8 salts, These sodium sails were prepared in situ from suberic acid {Tokyo Chemical Industry Co.) and sodium hydroxide.

Table 22A ON mHMh aaaaj ^^Octfedtid e ^monorSoa tiiin I berate^S?!· «IWZMim .^(ibareptide·-.·.,; Hydrophilic fraction API 0.058 0.059 PVP-12 2.6S0 2.620 mono-Sodium Suberate 15.087 0 di-Sodium Suberate 0 15.996 Water 0.712 0.747 Hydrophobic medium Span40 1.173 1.159 Lecithin 2.352 2.325 Ethyl isovalerate 10.169 10.055 Glyceryl monooleate 2.206 2.181 Glyceryl tributyrate 22.952 22.704 Castor oil 42.632 42.152 10 Die formulations described above in Table 22 are administered directly to the jejunum of non-anesthetized rats, and plasma octreotide levels are measured post formulation administration. Exposure values, ADC, are determined for the formulations and this is compared with a similar formulation prepared with sodium ocutnoate. 15 C - Geranic acid salt

Two octreotide-containing formulations were prepared essentially as described above wherein 12% sodium oetanoale was replaced by 18% geranic acid sodium salt (0.95M) and 14.6% (0.77M) geranic acid sodium salt, which is 3.7-dimethyl-2.6-ociadienoic acid 20 (obtained from SAHC.). The formulations produced are described in Table 22B below.

Table 22B - 1 - ;:'formufatiori;fAPU^z^;i ^Woctreotide^'^ i%fNaGerariat e!; ' -Nf! .’ds is:’· ?('Ti)Qttreotide·:· J 7 " NaGeranate - ,.i ft&amp;#943;&amp;#970;-&amp;#943;'· (% w) W pwS·' '•ι’&amp;#912; API 0.0S7 0.057 j Hydrophilic 1 NaOH 0 0.543 1 1 fraction j PVP 12 10.006 9.833 j j Sodium Geranate 18.053 14.625 J j Water 1.183 1.084 | j Tween 80 2-.001 i-970...J Hyd rophobic Glyceryl monocaprylate 4.001 3.923 J medium Glyceryl tricaprylate 63,235 65.927 j 1 Castor oil 0.000 o t 5 'fhe formulations described above in Table 22H were administered directly to die jejunum of non-unestheLized rats, and plasma octreotide levels were measured post formulation administration. Exposure values, AUC, were determined for the formulations and this was compared with a similar formulation prepared with sodium octanoate. 'Hie results are shown below in 'fable 22C and they demonstrate that the formulation with 1 K% sodium 10 geranate had similar activity as the 12%= sodium octanoate formulation, and the formulation with 14.6% sodium gcranate had increased activity.

Table 22C

Cargo formulation N AUC (0-601/dosc/kg h.w. ± SI) 15 Octreotide Sodium gcruiutte A 9 4.48 4 1.79 Sodium germane B 9 6.33 4 2.1 Improved 9 4.38 ± 1.66 20 Example 24: Effect of PVP (polyvinylpyrrolidone) on formulation activity 'Hie effect on formulation activity of replacing PVP-12 by mannitol (Sigma) was tested using formulations containing exenatide as cargo. It was understood in die art that PVP-12 is a stabilizer and could be replaced in the formulation by another stahilizer such as mannitol, fhc formulation shown in Table 23 below was prepared, This formulation is WO 2010/03214» ΡΟ71Β2009/»07155 73 a basic exenatide formulation prepared essentially as described above, but wherein PVP-12 is replaced by mannitol.

Table 23 Βββιι iTWBvfVTf ^FdrrhulatidhfAPi Pi · -;- 'i &amp;#943;^&amp;#970;. -7 *-· : ;:;®%'Hr»gredlent / ; Exenatide ? : .Mannitol ff T : (%w/wp ' Hydrophilic fraction API 0.055 tyigci; 0.137 Mannitol 2.742 Sodium octanoate 1.2.003 , MC400 0.137 Water 0.603 Hydrophobic medium Span40 1.213 Lecithin 2.434 Ethyl isovalerate 10.522 Giyceryl monooleate 2.2 S3 Giyceryl tributyrate 23.759 Castor oil 44,112 5 The formulation described above in Table 23 was administered directly io the jejunum of non-anesthetized nits, and plasma exenatide levels were measured post-formulation administration. Exposure values, AUC, were determined for the formulation compared to the basic formulation. These results are shown below in 'fable 24.

Table 24

If)

Cargo Formulation N AUC (0-60)/dose/kg b.w. ±SD Exenatide Basic 10 3.913.8 Mannitol instead of PVP-12 6 1.6 ±1.7

The results shown above in Table 24 demonstrate the surprising and unexpected 15 result tliat the exenatide formulation without PVP-12 had significantly decreased activity compared to the basic formulation. It was thus decided to investigate further the effect of PVP on bioavailability. 'Ihe effect on formulation activity of varying the molecular weight of PVP was tested using formulations containing exenatide as cargo. Three formulations of exenatide were 2(1 prepared using either PVP-12, PVP- 17 or PVP- 25 (all obtained from BASE). PVP- 12, PVP- 17 and PVP- 25 are all polyvinylpyrrolidone polymers; the average molecular WO 2010/032140 HCr/lB2009/«07155 74 weights arc about 2500-3000, 10000 and 30000 respectively. The formulations are basic exenatide formulations prepared essentially as described above wherein the PVP varies as described and wherein the hydrophilic fraction has been simplified to omit MgCband MC400. 5 Table 25 «ss SSBtt Λ'&amp;#938; ΡνΡ--12/17/25 * ,&amp;#943; Hydrophilic fraction API 0,022 PVP 12/17/ 25 2.752 Sodium octanoate 12.005 Water 0.602 Hydrophobic medium Span40 1.218 Lecithin 2.442 Ethyl isovalerate 10.551 Glyceryl monooieate 2.2S1 Glyceryl tributyrate 23.846 Castor oil 44.272

The three formulations described above in Table 25 were administered directly to the jejunum of non-anesthetized rats, and pi asi mi exenatide levels were measured post- 10 formulation administration. Exposure values, AUC. were determined for the formulations. The results are shown below in Tuhte 26.

Table 26

Cargo Formulation N AUC (0-60)/dose/kg b.w. ± SD (al Exfflytide PVP-12 1.1 8.0 ±7.7 PVP-17 δ 3,4 ±2.9 PVP- 25 5 2.δ ± 2.3

The results shown above in Table 26 demonstrate thui the exenatide formulations containing PVP- 12 showed much higher activity than the exenatide formulations 20 containing PVP- 17 and PVP- 25. Thus the effect of.PVP- 12 only was investigated further, and it was decided io perform a dose -response study using PVP- 12. The effect of increasing the amount of PVP- 12 in the formulation on the activity of the formulation was tested using formulations containing octreotide as cargo compound and different doses t'f PVP-12 as shown in Table 27 below. The PVP- 12 doses tested were 2,75% (the

WO 21U0/U3214U standard dose used in the above formulations) and 5.0%, 7.5%; and 10.0% PVP- 12; the hydrophilic fraction has been simplified to omit MgCl? and MC400. The formulation containing 10%· PVP was semi-solid i.e, it was apparently a semi-solid suspension.

Table 27 J' J^iForrnulatHonj^Pl^s1 . t - r \· T •bSrewentigEu^·;: litofii£'7SSCS$ ^Gctreotide^·^ ^PVP. 5io«'-;' i;>brtreptidfi"7 V PVP 7.5% : · .•.pctreptlile ) - RVP.iO.O%· '· i^(S6wZvi}^| iv) ’.v'? -- - ;'(%w/w} Hydrophilic fraction [ API 0.056 0.057 0.057 0.057 ] PVP-12 2.750 5,013 7.514 10.046 J 5odium octanoate 12.019 12.031 12.037 12.016 1 Water 0.593 0.6B4 O.7B4 0.685 ( Span40 1.217 1,183 1.145 1.108 t Lecithin 2.441 2.373 2.297 2.222 ( Ethyl isovalerate 10.554 10.259 9.934 9.608 Hydrophobic : Glyceryl monooleate 2.290 2.226 2.155 2.084 medium j Glyceryl tributyrate 23.832 23.166 22.431 21.694 | Castor oil 44,246 43.009 41.645 40.278 5

The formulations described above in Table 27 were administered directly io the jejunum of non-anesthetized rats, and plasma octreotide levels were measured post-foriuulalion administration. Exposure values. AUC, were determined for the four different formulations. 'Ihese results are shown below in 'fable 2KA. 10

Table 28A

Cargo Formulation N AUC (0-60)/dose/kg b.w. ± SD Octreotide 2.75% PVP-12 14 2.8 ± 1.0 5.0% PVP-12 12 3.7 ± 1.6 7.5% PVP-12 12 4.2 ± 1.5 10.0% PVP-12 11 4.7 ± 1.4 lhe results shown above in Table 28A demonstrate that the absorption of octreotide increased dramatically us die amount of PVP in the formulation increased. The WO 2010/0321441 PC'I7lB2009/007t55 76 formulation containing 10% PVP- 12 had absorption of octreotide about 1.7 times greater that the formulation containing 2.75%; PVP- 12, An improved octreotide formulation in which there. was 10 % PVP- 12 but no sodium octanoate showed virtually no activity, 'Hie rBA was 0.11 % (CV= 158¾) n=5, 5 It appears that the medium chain fatty acid salt acts as a permeability enhancer (by facilitating or enhancing permeability and/or absorption of die therapeutic agent 1, and that the PVP serves to increase the effect oflhe permeability enhancer in a synergistic manner since the PVP alone has virtually no effect. See also Example 31. A further experiment was performed to investigate if the 10% PVP-12 could be 10 replaced by dextran and still maintain activity of the formulation. Die dextran was manufactured by Fluka: the average molecular weight is -6000. The formulations were prepared essentially as described above wherein the PVP and dextran varies as described and wherein the hydrophilic fraction has been simplified to omit MgCEand MC400 and where the sodium octanoate was increased to 15%: see Example 26.

Table 28B ' : ' * J. ’ .· ; &amp;#970;Ε&amp;#912;&amp;#970;;&amp;#943;: A1' EE;/.:jngwdIent^^) VWfoCtre^ide^T::' ^vlOTtDtuttran'no'-P.VP ”···;. ' Octreotide- \ 5 ; SWDejrtramno PVP ' SiEiSi %wZwJ J: ^%w/w) Hydrophilic fraction API O.OS8 0.058 0.058 PVP-12 10.011 0,0 0.0 Dextran 0.0 10.011 5.011 Sodium octanoate IS .008 15.008 15.015 Water 1.003 1.003 0.803 Tween 80 2.027 2.027 2.169 Hydrophobic medium Glyceryl monocaprylste 4.036 4.036 4.319 Glyceryl tricaprylate 40.714 40.714 43.574 Castor oil 27.143 27.143 29,049

The three formulations described above in Table 2SB were administered direct])’ to the jejunum of non-anesthetized rats, and plasma octreotide levels were measured post- 20 WO 201()/03214(1 PC’I7IB2(IO9/OO7155 77 formulation administration. Exposure values, AUC, were determined for the formulations. 'Hie results are shown below in Table 28C.

Table 28C

Cargo Formulation 1 N AUC{h-25t/d»xc/kg b.w.±SI> I0%PVP i 9 4.4 ± 1.7 Octreotide 10% Dextran; no PVP J 5 3.3 ± L.h 5% Dextran; no PVP 1 9 3.2 ± 1.5 5 lhe results shown above in Tabic 28C demonstrate lhai die absorption of octreotide decreased when PVP in the formulation was replaced by dextran but the activity was still significant. The formulation containing 10% dextran had absorption of octreotide about 15% of the formulation containing 10% PVP, and the formulation containing 5% dextran had absorption of octreotide about 13% of the formulation 10 containing 10% PVP.

Example 25: A comparative study of C8. C9 and CIO medium chain fatty acid salts viz., sodium octanoate, sodium nonanoate and sodium decanoate 'Hie effect on formulation activity of replacing sodium octanoate wiLh other 5 medium chain Fatty acid sodium sales was tested using formulations containing octreotide as cargo. Three formulations of octreotide were prepared, as shown in Table 29 below, These are all basic formulations prepared essentially as described above where the hydrophilic fraction has been simplified to omit MgCb and MC40O and wherein the medium chain fatty acid salt is an equimolar amount of sodium octanoate. sodium 10 nonanoate or sodium decanoate.

Table 29 iiH |^F6muiaif MB &lt;^Ν&amp;#943;&amp;#943;€8512«Μ ^'Gctreotidd^ »&lt;α·72Μ)&amp;amp;Ο, ^Octreotide,'; j ®?(&amp;amp;72Μ)&amp;#943;Μ;&amp;#943; 'β!κ72Μ)β API 0.058 0.057 0.058 Hydrophilic fraction PVR-12 2.750 2,718 2.585 Sodium octanoate 12.019 0 0 Sodium nonanoate 0 13.023 0 Sodium decanoate 0 0 14.019 Water 0.593 0.632 0.670 Span40 1.217 1.203 1.188 Lecithin 2,441 2.412 2.383 Ethyl isovalerate 10.554 10.428 10.303 Hydrophobic Glyceryl monooleate 2.290 2.252 2.235 medium Glyceryl tributyrate 23.832 23.547 23.265 Castor oil 44.245 . ' 43.718 43.194 'Hie fonnulations described above in Table 29 were administered directly to the 15 jejunum of non-anesthetized rats, and plasma ocueoiide levels were measured post- formulation administration. Exposure values. AUC, were determined for the formulations. The results are shown below in Table 30. WO 20111/032140 79

Table 30 J Cargo Formulation N AUC (0-25)/dose/kg b.w. ± SD Octreotide sodium octanoate NaC8 9 2.1+0.8 sodium nonanoate NaC9 10 2.5+0.4 sttdium decanoate NaClO ' 10 1.7 + 0.4 5 'lhe results shown above in 'fable 30 demonstrate that when sodium ocianoate in the formulation is replaced by sodium nonanoate or by sodium decanoute there is similar activity. Based on statistical analysis, there is no difference in activity between ait three formulations.

Example 26: Dose response of sodium octanoate 10 lhe dose response of sodium octanoate at 12%, 15% and 18%: was tested by making the formulations shown in Table 31. These are all basic formulations prepared essentially as described above where the hydrophilic fraction has been simplified io omit MgGb and MC400 and die cargo compound was octreoLide. Additionally the formulation was corrected for viscosity i.e. the same or similar viscosity was maintained for all three 15 formulations; this was achieved by varying the amounts of castor oil and glyceryl tributyraie.

Table 31 V ' &amp;#970;&amp;#943;* p.Formulation, API/h ip^Octfeotid iiTpctreciUde·.-T-NaC8l5% :T .-Octreotide ' NaC818A / ·, ’(%w/w) J Hydrophilic fraction (simplified) API 0.058 0.058 0.058 PVP- 12 2.750 2.652 2.554 Sodium octanoate 12.019 15.040 18.016 Water 0.593 0.710 0.825 Hydrophobic medium Span40 1.217 1.173 1.130 Lecithin 2.441 2.353 2,267 Ethyl isovalerate 10.554 10.175 9.802 Glyceryl monooleate 2.290 2.207 2.126 Glyceryl tributyrate 23.832 32.816 41.090 Castor oil 44.246 32.816 22.132 WO 201(1/032140 P€’i7l82t)t)9/fH)7155 80

The formulations described above in Table 31 were administered directly lo the jejunum of' non-anesthetized nils, and plasma octreotide levels were measured post-form ulation administration. Eixposure values, AUC. were determined for the formulations. The results are shown below in Table 32.

Table 32

Cargo Formulation N AUC (0-60)/dose/kg b.w. ± SD Octreotide NaCH'12% ' 14 2.8 ± 1.0 NaCS 1'5% 12 4.1 ±1.9 5f) NaC8 18% 12 3.6 ± 1.1

The results shown above in Table 32 demonstrate that when sodium octanoate in the formulation is increased from 12% to 15% there is an increase in activity but a further increase of sodium octanoate to 18% leads no higher activity than that obtained at 15%.Thus about 15% sodium octanoate appears to be the preferred amount.

Example 27: Investigation of the effect of varying the hydrophilic /lipophilic balance of the surfactants in the formulation

Table 33 below describes various octreotide formulations. 'Hie first column, formulation (a), is Lhe basic formulation prepared essentially as described above, where the hydrophilic fraction has been simplified to omit MgCb andMC4(K). and the cargo compound is octreotide. The surfactants are Span 4(1, lecithin anti glyceryl monooieate. and by calculation the ΕΓΕ.Β is approximately 5-6. In the other formulations (formulations h, c. and d) the HLB was changed ax indicated (to 3,5. 6,7 and 14) by replacing Span 40 and lecithin by differing amounts of Tween 80 and by varying the amount of glyceryl monooieate. WO 2010/1)32140 PCI71B21HWUU7155 81

Table 33 · i'Jb'^frriutattohpARI^ :.· :[ &lt;: Ingredient ^Octreiotidafi Μ; HLB5-6: [a 1 ^Octreotide'^ F^Ottfetftide- "£ A' T-«Octreotide I; - : HLfl,14[d]‘:% "’· (%w/w) ;{%w/w) 3 Hydrophilic fraction API 0.058 0.057 0.057 0.057 PVP-12 2.750 2.748 2.748 2.748 Sodium octanoate 12.019 12.027 12.027 12.027 water 0.593 0594 0594 0.594 Hydrophobic medium '5pan40 1.217 0 0 0 Lecithin 2.441 0 0 0 Ethyl isovalerate 10.554 10.547 10.546 10547 Tween 80 0 0502 2.003 5500 Glyceryl monooteate 2.290 5500 4.002 0.502 Glyceryl tributyrate 23.832 23.811 23.811 23.811 Castor oil 44.246 44.215 44,215 44.215

The formulations described above in Table 33 were administered directly to the 5 jejunum of non-anestheiized rats, and plasma octreotide levels were measured post- ibmiulation administration. Exposure values, AUC, were determined for the formulations. The results are shown helow in Table 34. 10 15

Table 34

Cargo Formulation N AUC f0-25)/dose/kg b.w, ± SI) Octreotide HLB 5 la] 9 2.1 ±0.8 HLB 3.5-fb] 12 3.3 ±0.9 HUB 6.7 — [cl 11 3.8 ±0.9 HUB 14 - [dj 10 3.7 ±0.9 20

The results shown above in Table 34 demonstrate that all the three new formulations replacing Span 40 and lecithin with Tween 80 [h. c and dj had much belter activity than the basic formulation [al. even although the HUB in [b] was lower, in |c| was slightly higher and in [dl was much higher than die HUB of die surfactants in (a). Additionally, the activities of all the new formulations |(b, c, and d] were statistically very similar. Thus the HUB alone of the surfactants does not seem to affect activity but the characteristics of the surfactants appear to play an important role. In particular, replacing Span 40 and lecithin with Tween 80 is advantageous for activity in these octreotide formulations. 25 WO 2010/03214(1 82

Example 28: Octreotide formulations with different ratios of glyceryl tricaprvlate to castor oil.

Based on die accumulation of results described above including the PVP- 12 dose response results, the sodium octanoate dose response results and the surfactant results 5 inter alia, a series of octreotide formulations were prepared using 10% PVP- 12 and 15% sodium octanoate. and varying the ratio of glyceryl tricaprvlate κ.» castor oil. Additionally, glyceryl inonooleaie and glyceryl tri butyrate were replaced (if used) by glyceryl inonocaprylatc and glyceryl tricapryiatc (both supplied by Abitec). 'Hits is io maintain'the CK motif within the formulation. 'Hius the hydrophilic fraction contains a salt of a C8 acid 10 (ocianoaic) and the hydrophobic medium contains monoglvcerides and triglycerides incorporating the same CS acid. 'Hie inventors believe that the use of C-8 compounds in both the hydrophilic fraction and in the hydrophobic medium may be advantageous for hioavailability. 'Hie amounts of Tween 80 and glyceryl monocaprvlaie were also varied in the formulations. 'Hie formulations were prepared are shown in 'Table 35A below, 15 Formulations 1. II. V and VI were semi-solid (apparently suspensions! and formulations ΙΠ and (V were the usual liquid suspensions.

Table 35A . L -ς . ’ J, ' r j. vwGs Ltrdrmui^iohXARtjia «Octreotide «Octreotide «Octreotide 'Vsiv-fcz^i ^Octreotide; <ji Octreotide « /{Kw7wJ>; . ; , Hydrophilic fraction API 0.058 0,058 0,058 0.058 0.058 0.058 PVP-12 10.011 10,011 10.011 10.011 10.011 10.011 Sodium octanoate 15.008 15.008 15.008 15.008 ' 15.008 15.008' Water 1.003 1.003 1.003 1.003 1.003 1.003 Hydrophobic medium Tween 80 2.027 2.027 2.027 2.027 6,063 6.062 Glyceryl monocaprylate 4.036 4.036 4.036 4.036 0 0 Glyceryl tricaprylate 40.714 13:571 61.071 67.857 40.714 0 Castor oil 27.143 54.286 6.786 0.000 27.143 67.857 20 'Hie formulations described above in Table 35A were administered directly to the jejunum of non-anesthetized rats, and plasma octreotide levels were measured post-

formulation administration. Exposure values. AUC, were determined for lhe fomiulaiiuns. llie results are shown below in 'fable 35B.

Table 3SB

Cargo Formulation N AUC (0- 25)/dose/kg b.w. + SD Formulation 1( GTC xastor oil 6:4) 9 4.4 ±1.7 Formulation ll(GTC xastor oil 2:8) 8 3.0+1,7 Octreo- tide Formulation HI (GTC xastor oil 9:1) 9 3.1+0.5 Formulation !V(GTC xastor oil 10:0} 7 4.1 ±2.1 Formulation V -without GMC (GTC xastor oil 6:4) 6 1.6 + 1.0 Formulation VI -without GMC&amp;amp;GTC (GTC xastor oil 0:10) 7 1.1+0.6. lhe results shown above in Table 35B demonstrate that formulations 1 and IV have greatest activily, Since castor oil is absent in formulation IV tilts demonstrates that castor oil is not essential for activity. It seems that a high GTC: castor oil ratio e.g. 6:4 is beneficial for activity. Additionally, since formulation V (which has low activity) has the 10 same GTC: castor oil ratio as formulation I it appears that additionally CMC (or other monoglyceride) is desirable for activity. Additionally a formulation similar to formulation 1 of fable 36 was prepared but sodium octanoate was omitted. This formulation showed virtually no activity. rBA=0.1 %.

Bulk drug product of formulation IV (improved, no castor oil) was milled with a I 5 150 micron screen, and then particle size was determined using Malvern Iraser 1) i ffracti on t cch nology. Pre li min ary results i ndicated that 90% (v/v) of die parti cl es w ere below 130 microns, and 50% (v/v) of the particles were below 45 microns.

Iheltminary experiments using similar formulations io formulation 1. but with varying increased amounts of octreotide all gave similar BA i.e. there was approximately 20 linear exposure independent of API loading. Λ preliminary experiment using a similar formulation to formulation IV at even higher octreotide loading - 1.5% (wl/wt) - also gave similar ΒΛ. A similar improved formulation to formulation 1 above was prepared using HD4 as cargo instead of (xlreotide, and it was compared to a basic formulation. These 25 formulations are described in Table 36A below. WO 2010/03214« 84

Table 36A - '· /TTj.’rf MHR .TiBauc (noMg/M Μη» «itw^flrnprowed^i:· '7 j.tSiiiitSSWfe'hiM-iS.i-S;·:· · ifif'·' ii-JiiXA:! it HydroDhtiic fraction API 0.545 0.545 NaOH 0.001 0 PVP-12 2.734 10.012 Sodium octanoate 12.036 15.009 Water 0.613 1.023 Hydrophobic medium Tween 80 0 2.013 Glyceryl monocapryiate 0 4,008 Giyceryl tricaprylate 0 40.434 Span40 1.21 0 lecithin 2.42 0 Ethyi-lso-vaierate 10.49 0 Giyceryl mono-oleate 2.28 0 Giyceryl tributyrate 23.69 0 Castor oil 43.98 26.956

The formulations described above in Table 36A were administered directly to the jejunum of non-anesthetized rats, and plasma HD4 levels were measured post- formulation 5 administration. Exposure values, AUC. were determined for the formulations. 'ihe results are shown below in Table 36B.

Table 36B I Cargo 1 Formulation N AUC (0-901/duse/kg b.w, ± SD 1 HD 4 (dextran) Basic 6 67448+ 16977 Improved 6 95374 + 47490 10 The results shown above in Table 36B demonstrate that the improved formulation has much greater activity than the basic formulation.

Example 29: Detailed production process for a selected (improved) octreotide formulation

Ihe octreotide formulation in Example 28 (Table 6, first column) was prepared 15 essentially as described in the above Examples. Below follows the detailed production process for this formulation. WO 2010/032140 !&amp;#908;7ΙΒ21ΙΟ9/&lt;ΗΙ7Ι5$ 85

Production of the hydrophilic fraction:

To 150 ml-, water the following ingredients were slowly added and mixed: 24.05 g of sodium octanoate, 16.04 g of PVP-12 and 92.4 g of 10 mg/mL aqueous octreotide solution. The resulting solution wus lyophilized. 5 Production of the hvdrophohic medium: 3.25 g Tween 80. 6.47 g of glyceryl monocaprylate, 65.25 g of glyceryl trieaprylate and 43.50 g of castor oil were mixed together.

Production of the bulk drug product: . 26.08 g of the hydrophilic fraction was slowly added to 73.92 g of the 10 hydrophobic medium at 20±2 "C while mixing. After addition of the entire hydrophilic fraction, the mixing speed was increased. Degassing by vacuum was dien applied and the resulting suspension was stored at 2-8°C.

To enahle larger amounts of octreotide to he dissolved die following method was devised: 15 1. ’Die amount of water of the hydrophilic fraction preparation was the same as the calculated volume of die final bulk drug product, 2. PVP-12 was dissolved in half of the above amount of water. 3. Sodium octanoate was dissolved in the second half amount of water. 4. Octreotide was dissolved in die PVP-12 solution (from paragraph 2). 2() 5. The sodium octanoaie solution was added to the octreotide and PVP-12 solution.

At this stage there was some precipitation, but it became soluble after mixing.

Example 30: Experiments in pigs using capsules 25 In order to test the activity of the formulations of the invention when administrated in capsules, an animal model allowing capsule administration to pigs (domestic swine) was established. In order to bypass the stomach and allow direct administration of capsules to the small intestine of the pig. a well established model in dogs {“Nipple Valve model**: Wilssun-Rahniberg &amp;amp; O. Jonsson, Laboratory Animals 30 (1997), 31. 231 -240) was adapted lo the commercial pig.

The two octreotide formulations shown below in 'fable 37 were prepured, 'lhe octreotide (x) formulation was prepared essentially as described above lor the basic formulation wherein die hydrophilic fraction has been simplified to omit MgCb and MC400. 'fhe octreotide (y) fonnulation was prepared essentially as described above for WO 2« 10/032140 lhe improved octreotide formulation. The formulations were filled into gelatin capsules (fn>iu Capsugeh, basic formulation(x) at 0.42 mL/capsule and improved formulation (y) at 0.44ml7capsulc. resulting in 5nig net octreotide content in both types of filled capsules. 'Hie capsules were not enteric-coated i.e. they were uncoaled.

Table 37 aMtil ^Octrebtide:(x): y •^pttfeotideiy);;/ &lt;.77fitnpr0ved.?.&lt; Hydrophilic fraction API 1.357 1.277 PVP-12 2.717 10.011 Sodium octanoate 12.011 15.00B Water 0.643 1,052 Hydrophobi c medium Tween 80 0 1.992 Glyceryl monocaprylate 0 3.967 Glyceryl tricaprylate 0 40.016 Castor olt 43.562 26.677 Span40 1.198 0 Lecithin 2.403 0 Ethyl isovalerate 10.391 0 Glyceryl monooleate 2.254 0 Glyceryl tributyrate 23.463 0

The formulations described ahovc in Tuble 37 were administered directly to the 10 small intestine of the non-anesthetized pigs via the gastric bypass described above, and plasma octreotide levels were measured post-administration. Exposure values. AUC were determined for the formulations. The % BA was calculated compared to the exposure to octreotide after subcutaneous administration. ITe results obtained are shown below in 'fable 38. 15

Table 38

Cargo Formulation N AUC (0-240) ± SD | % SA± SD Octreotide Octreotide(x) 4 896 ± 305 j 2.1 ±0.7 Octreotide(y) 4 2574 + 889 1 6.2 ±2.1

The above results in Table 38 show that there was bioavai lability in the pig model for encapsulated formulations, for both the basic and improved formulations. Octreotide 87 bioavai lability of the improved formulation was about three times the level of bioavai lability of the basic formulation. 'Ihe results given here for bioavailability are underestimated because sampling time was not sufficient for octreotide levels to go back to baseline (0 ng/niL). This was 5 due to the unexpectedly longer exposure time in pigs as compared to what had been previously measured in ruts, The shape of the graph was changed compared to the rat results show'ing longer lime to reach maxi nt al peak levels and extended time in which octreotide is resident in the blood. This may he advantageous since this allows the octreotide to he longer-acting in the body. 'Ihus the actual bioavailability in pigs must be 10 higher than the numbers given.

Based on the results in rats, the level of bioavuilabilitv in pigs of octreotide administered in aqueous solution is extrapolated to be about 0.1 %. This level of bioavailability is below' the level of sensitivity of the bioassay used for pigs. 15 Example 31: Dose-response results for PVP in the Improved Formulation.

Further to the PVP results in Example 24. the effect on activity of increasing die amount of PVP-12 in Lhe improved formulation was studied. The improved formulations, made essentially as described above, contained octreotide as cargo compound and different doses of PVP-12 as shown in Table 39 below. The PVP- 12 doses tested were 20 7.5%. 10.0% and 15.0% PVP-12. The formulations containing 10% and 15.0% PVP were semi-solid i.e. they were apparently semi-solid suspensions- and the formulation containing 7.5% PVP was a viscous suspension.

Table 39 ‘7 p ' f-z Vil Formulattoni'API sasaWBw Wdctreotke tamfe ^QdtnwtWe ++3 % -Octreotide. Hydrophilic fraction APS 0.058 0.058 0.058 PVP- 12 7.SO6 10.011 15.009 Sodium octanoate 15.012 15.008 15.009 Water 0.903 1.003 1.203 Hydrophobic medium Tween 80 2.098 2.027 1.884 Glyceryl monocaprylnte 4.178 4.036 3.752 Glyceryl tricaprylate 42.147 40.714 37.851 Castor oil 28.098 27.143 22.234 WO 20111/032140 88

The formulations described above in Table 39 were administered directly to the jejunum of non-anesthetized rats, and plasma octreotide levels were measured post-formulation administration. Exposure values. AUC, were determined for the three 5 formulations. These results are shown below in 'fable 40.

Table 40

T

Cargo Formulation N AUC (0-25)/dose/kg b.w. + SD Octreotide 7.5% PVP-12 7 2.9 + 2.2 10O% PVP-12 9 4.4 + 1.7 15% PVP-12 10 2.1 + 1.2 lhe resuIls shown above in Table 40 demonstrate that the absorption of octreotide was greatest when PVP in the formulation wus 10%. and increasing the amount to 15% results 15 in significant decrease in activity. This confirms the choice of 10% PVP in the improved formulation.

Experiment 32: Activity of API packed in formulation compared to API administered concomitant to formulation 20 Three different basic formulations of three different cargo compounds were prepared (dextran, gentamicin and exenatide), essentially as described above (wherein the basic formulation is die basic non-simplilied hydrophilic fraction). Each of these three formulations was administered diiecdy to the jejunum of non-anesthetized ntus. and plasma cargo levels were measured post- formulation administration. Exposure values, 25 AUC. were determined for the formulations. Additionally, a similar formulation was prepared with a non-relevant cargo compound (a mock formulation). Separately, the mock formulation was administered concomitantly with dextran, gentamicin or exenatidc in aqueous solution and exposure vulues, AUC, were determined. Concomitant administration was achieved by administrating cargo in aqueous solution immediately 30 followed by mock formulation administration via a jejunal- implanted cannula (gastric bypass). For each compound, exposure after administration of die formulated cargo was compared to exposure after administration of the unfonuulated cargo (concomitant), lhe WO 2010/032141) comparative results are shown below in Table 41. The results show that there is higher activity (bioavailability) when the cargo is formulated compared to unformulated (concomitant) in all three cases, and that exenatide showed by far the greatest increase in activity dueto formulation. Note that dextran and gentamicin are compounds that are not 5 sensitive to protease degradation, whereas exenatide being a peptide is subject to degradation by intestinal enzymes. The large difference in aetivity between the formulated exenatide compared to unformulated exenatide may be due to the protective effect of the formulation against degradation.

Table 41 APl/cargo Formulated versus unformulated (fold activity) Dextran 1.7 Gentamicin 1.5 Exenatide 4.4 10

Example 33: Intestinal hyperpernieabiiity evaluation A. Size limitation: The technology and formulations described above are

intended to enhance the permeability of the intestine, allowing specific deliver;' of proteins, peptides and other otherwise impermeable molecules across this barrier. A 15 certain degree of non-specific penetration of intestinal content may result as a side-effect of this enhancement of specific permeability. The size of molecules which could possibly penetrate the intestine in a non-specific manner was evaluated using different molecular size markers. hi order to evaluate the molecular- size limit of increased (11 permeability, five 20 different HTC-labeled dextrans of different molecular weight were chosen to serve as molecular markers to lest increased intestinal permeability: the average molecular weight of the five dextrans was 4.4. 10, 20, 40 and 70 kDa. equivalent to a radius of 14. 23, 33. 45 and 60 A respectively. These different size markers were administered directly to the jejunum of non-anaesthetized rats, through an intestinal implanted cannula, and showed 25 virtually no basal intestinal penetration when tested alone. Each of these markers was then administered directly to the jejunum of non-ancsthetized rats together with 300pL of WO 2011)/()3214(1 ΚΤ/ΙΒ2009/1Μ)7Ι55 90 basic formulation. and the degree of its penetration was evaluated by testing dextran levels in blood.

The plasma dextran levels were measured pre-dosing and at 3'. 6', 10'. 25’. 60'. 90' minutes post formulation administration. Exposure values. AUC (0-90). were 5 determined and the results are shown in Figure 7. Data is presented as MEAN ± SD. n>4. lhe results show that while the smallest molecular marker tested (dextran of average MW=<4 kDa). penetrates the intestine when administered concomitantly with a formulation, as the molecule size increases, penetration extent decreases: a marker, molecule of 10 kDa penetrates to a smaller extent and a 20 kDa marker to an even smaller 10 extent. Λ marker molecule of 40 kDa shows minimal penetration, while a marker molecule of 70 kDa shows no penetration al. all (basal penetration). These results indicate that 40-70 kDa is a cutoff size for non- specific permeability enhancement by formulations οΓ the invention. Thus administration of a large volume of fonnulation (300pL) to the jejunum of rats resulted in permeability enhancement of the intestinal 15 barrier, and this enhanced permeability is restricted by molecular size, showing a cutoff size of 40-70kDa and minimal penetration at 40kDa.

Published values of the size of hazardous molecules (molecular weight and radius) which could potentially he present in the intestine are shown below in Table 42.

Table 42

MW Q ) Radius (A) Macro moloculos i A o r I arg er LPS » 1 oo , Stno rt - 1 OO Long - 1 OOO Enterobacterial Toxins 70- 900 - Viruses - eoo - i ooo | Bacteria - 1 0,000 or | larger H

Table 42 demonstrates that potentially hazardous molecules present in die intestine are above the cutoff size of permeability enhancement by the tested formulations. as shown above. 'JTius these results suggest that die tested formulations will 25 not facilitate penetration of hazardous molecules through the intestinal barrier anti these WO 2010/03214» formulations can therefore be considered as safe. Other formulations of the invention give simitar results. B. Formulation repeated dosing: In order to investigate if repeated dosing of formulation affects intestinal permeability, the octreotide improved formulation (12% 5 sodium oetanoale with cuslor oil) was dosed to rats for 14 sequential days using the above in vivo model (rat implanted with two cunnulas in die jejunum). At days 1.7 and 14 of administration, a dextran permeability marker (FITC-dextran of 4.4 kDa MW: FD41, was administered 60 minutes post formulation administration. This was to assess the permeability of the intestine by the penetration of the FD4 from the intestine to blood. No 10 significant difference in FD4 exposure following 14 days of formulation repeated dosing was found. 'These results suggest there is no increase in intestinal permeability following this period of repeated dosing of formulation, and intestinal enhanced permeability remains a reversible process during this period. 'Hie results suggest that the formulation causes no damage to the intestinal tissue. 15 hut acts by specifically opening the intestinal barrier, showing no additive permeability enhancing effect.

Example 34: Intestinal hyper permeability evaluation: time-course and reversibility

Further to the study in the above Example, a study was designed in order to define 20 the time-course of increased intestinal permeability due to Lhe formulations of the invention, and the reversibility of this process, using dextran as a permeability marker. in Older io define the time window of increased intestinal permeability, an in vivo model was developed in rats, in which one or two cannulas are implanted in the jejunum of the rats. HTC-laheled dextran (average molecular weight 4.4kDa. FD4), which has 25 virtually no basal intestinal penetration, served as a molecular marker to test intestinal permeability. An experiment was designed in which foe dextran marker was administered concomitant io Lhe formulation (by a jejunal implanted cannula), or at different time intervals from the formulation administration (by a second separate jejunal implanted cannula), Intestinal permeability was evaluated by testing FD4 penetration to blood. Rats 30 were administered a basic formulation concomitant with the dextran marker, or the basic formulation and then the dextran marker at different intervals of time (10. 30 and 60 minutes). Blood samples were analyzed for dextran concentration pre-administration and WO 2010/1132140 FCT/IB2009/IMI7155 92 at 3. 6. 10, 25, 60 and 90 min following dextran administration. The results are shown in Figure K. Data is presented as Mean ± SD, n>5.

Figure 8 demonstrates dial the dextran marker penetrates the intestine to the highest extent when administered together with the formulation. An interval of 10 5 minutes between administration of the. formulation and administration of the dextran marker results in significantly decreased amount of murker penetration, and increasing the interval further results in exponential reduction of marker penetration.

These results show that while there is some degree of non-specific permeability enhancing by die formulation, it is restricted to a short period of lime following 10 administration of die formulation. The permeability of the intestine decreases sharply with time, and 60 minutes from administration of the formulation diere is no more marker jieneirution. Thus administration of the formulation to the rat intestine results in a very' short period of hyperpenneability of the intestinal barrier. Other formulutions of the invention gave similar results. 15 Example. 35: Oral administration of octreotide to monkeys

In order to test the pharmacokinetics of octreotide following oral administration of formulated octreotide to monkeys, five Cynomologus monkeys were orally dosed with capsules containing an improved castor oil formulation of octreotide (similar to formulation 1 of Table 35 - but with higher load of octreotide). 'Hie capsules used were 20 size 1 gelatin capsules coated with 6.7% Acryl-EZE® enteric coating; tliis coating prevents capsule disintegration in the stomach and allows opening of the capsules in the small intestine of the dosed animals. The octreotide dose used was 5mg/capsule.

Monkeys were fasted overnight prior to capsule administration, Following oral administration, blood samples were withdrawn over a period of 9.75 bouts, processed for 25 plasma and analyzed for octreotide content bj' the LC/MS/MS method: see Figure 9. Similar experiments were performed with the improved no castor oil/ GTC formulation (simitar io formulation IV of Table 35 but with higher load of API) and similar results were obtained. Similar experiments were also performed with several different enteric coatings and similar results were obtained. 30 In order to compare the pharmacokinetics of octreotide following administration of the improved octreotide formulation, to the pharmacokinetics of injected octreotide, octreotide acetate solution (0.1 mg/ monkey) was administered subcutaneously to two monkeys from the above group to serve as a reference. Blood samples were withdrawn WO 2011)/03214(1 over a period of four hours, processed for plasma and analyzed for octreotide content by the tC/MS/MS method.

The pharmacokinetics of oetreoLide following oral octreotide and subcutaneous injected octreotide solution were compared (see Figures 9 and 10(. The results of the oral 5 formulation showed absorption over a period of a few hours. The shape of the graph was changed compared to subcutaneous, showing slower but longer release of octreotide into the blood, 'litis may be advantageous since this allows die persistence of octreotide for a longer lime in the blood potentially prolonging the activity window. '

An approved dose- for injected octreotide acetate in humans is 0.1 mg/ patient. Ihc 10 above results in the monkeys suggest that the improved formulation containing about 10 mg octreotide per dose will generate therapeutic exposure in humans.

Example 36: Stability data

Basic and improved octreotide formulations of the invention were maintained 15 both at 4 UC and at 25 °C and were tested for octreotide content periodically. Both formulations were found to be stable.

Example. 37: Formulations incorporating vancomycin, interferon-alfa and terlipressin 20 A. Vancomycin; Tabic 43 below describes a vancomycin improved formulation. containing 10% PVP and 15% sodium octanoate in the hydrophilic fraction, and containing glyceryl tricaprylatc as the main constituent of die hydrophobic medium. The vancomycin was obtained from Gold Biotechnology.

Table 43

Formulation, API Ingredient Vancomycin (%w/w) Hydrophilic fraction API 6.267 NaOH 0.082 PVP-12 10.005 Sodium octanoate 15.016 Water 1.216 Hydrophobic medium Tween 80 2.004 Glyceryl monocaprylate 4.008 Glyceryl tricaprytate 61.400 Castor oil 0.000 WO 2010/03214(1 94

In a preliminary experiment, the formulation described ubove in Table 43 was administered directly to the jejunum of non-anesthetized rats, and plasma vancomycin levels were measured post- formulation administration. Exposure value. AUC, was 5 determined for liie formulation. The results are that the absolute BA is around 5% (comparative to TV. n-6). When vancomycin in saline solution wus administered to the jejunum of non-anestheLized rats no BA was detected. liiterfei'on-nlfa: Table 44 below describes an interferon-alfa improved formulation, containing 10% PVP and 15% sodium octanoate in Lhe hydrophilic fraction, and 10 containing glyceryl tricapryiate as the main constituent of the hydniphohic medium. The interferon-alfa is supplied in a buffer (from int as Biophamiaceuticais) and the ingredients of the interferon-alfa buffer in the formulation are marked by an asterisk (*).

Table 44

Formulation, API Ingredient IFN-o (%w/w) Hydrophilic fraction API 0.Q50 •Na;HPO4 0.032 *NaHjPO&lt; 0.030 ‘Polysorbate (Tween) 80 0.002 ‘Disodium EDTA 0.002 PVP-12 10.025 Sodium Octanoate 14.997 Water 1.006 Hydrophobic medium Tween 80 2.005 Glycery) monocaprylate 4.005 Glyceryl tricapryiate 67.84 Castor oil 0 15 Phe formulation described above in 'fable 44 is administered directly to the jejunum of nnn-anesthetized rats. Plasma interferon-alfa levels are measured post-formulation administration. C. Terlipressin: 'fable 45 below describes a lerli press in basic formulation and a teriipressin improved formulation containing 10% PVP and 15%· sodium octanoate in Liie 20 hydrophilic fraction, and containing glyceryl tri caprylate as the main constituent of the hydrophobic medium. The terlipressin was obtained from Bambio. 'Phe basic formulation 95 was prepared essentially as described above and die improved formulation is also prepared essentially as described above.&amp;#906;

Table 45 / if :(=¾ ,· ti . -.z ,0. · >>v;Forrnulation,>APIp;. erlipressini T basic^iyy, '.^yTeflifirMsin y\ fS:3improtfed **<$<<&amp;*·**$ sssee Hydrophilic fraction API 0.235 0.235 MgCl, 0.137 0.000 PVP 12 2.735 10.004 Sodium octanoate 12.004 15.015 MC 400 0.137 0.000 Water 0.610 1.010 Hydrophobic medium Span40 1.211 0.000 lecithin 2.428 0.000 Ethyl isovalerate 10.500 0.000 Gtyceryl monooleate 2.278 0.000 Glyceryl tributyrate 23.708 0.000 Castor oil 44.016 0.000 Tween 80 0.000 2.002 GMC 0.000 4.004 GTC 0.000 67.731 'Phe formulations described above in Table 45 are administered directly to the ! 0 jejunum of non’anesthetized rats. Plasma terlipressin levels are; measured post-formulation administration.

Example 38: Inhibition of growth hormone in vivo by octreotide

One of die best characterized effects of octreotide is the inhibition of growth hormone release. In order to test for the efficacy of an octreotide formulation of the invention on 15 growth hormone inhibition, a rat model was used in which endogenous rat growth hormone (rGH) levels were monitored following octreotide formulation administration io the jejunum of the non·anesthetized rat model (described above). Administration of a basic octreotide formulation (containing 12% sodium octanoate) to the jejunum of rats was shown to reduce rGH levels by 87.4% compared to administration of a saline control, WO 2010/03214(1 PC'I71B200y/007155 96 'Hiis result demonstrates dial the octreotide formulations described herein enable deli very of octreotide in its active form from the intestinal lumen into the blood stream. 5 Example 39: Toxicology studies Λ 28-day toxicity administration study of fonnutation control (excipiunis only, no cargo) was performed in Wistar rats. The animals in die lest group were daily administered rectally witii the maximal feasible dose of formulation (100 pL/animal/day) for 28 1(1 consecutive days. The lest group was compared to two control groups: a naive group (non- treated) and a saline administered group, (n= 1.5/ group).

General clinical observations were made twice daily, and detailed clinical observations were performed weekly. Body weight and food consumption were measured weekly. Clinical pathology and gross pathology were conducted one day after the lust 15 treatment. Λ histological examination was performed on rectum, colon, liver and kidneys, and no toxic effects were detected. There was clean histopathology with no local Gl or systemic Findings, no formulation related clinical findings, no changes in hematological and blood chemistry parameters, no macroscopic findings at necropsy and no mortality, in conclusion, this experiment demonstrated that there was no observed toxicity during a 20 daily recud dosing of formulation to nils for 28 consecutive days. I laving thus described several aspects of at least one embodiment, it is to be appreciated that various alterations, modifications, and improvements will readily occur to those skilled in the art. Such alterations, modifications, and improvements are intended 25 to be pan of this disclosure and are intended to be within the scope of the invention.

Accordingly, the foregoing description and drawings are by way of example only, and the scope of the invention should be determined from proper construction of the appended claims, and their equivalents.