Enantiomers of spiro-oxindole compounds and their uses as therapeutic agents.

(54) Title: ENANTIOMERS OF SPYROOXINDOL COMPOUNDS AND THEIR USES AS THERAPEUTIC AGENTS. (54) Title: ENANTIOMERS OF SPIRO-OXINDOLE COMPOUNDS AND THEIR USES AS THERAPEUTIC AGENTS.

(57) Summary

This invention is directed to the (S) -enantiomer of the compound of formula (I): (See Formula) or a pharmaceutically acceptable solvate or prodrug thereof; This (S) -enantiomer is useful in the treatment of diseases or conditions, such as pain, that are ameliorated or alleviated by modulation of voltage regulated sodium channels.

(57) Abstract

This invention is directed to the (S) -enantiomer of the compound of formula (I) or a pharmaceutically acceptable solvate or prodrug thereof. This (S) -enantiomer is useful for the treatment of diseases or conditions, such as pain, which are ameliorated or alleviated by the modulation of voltage-gated sodium channels.

_I KNOW_

StCMWÍA I heard ECONOMY

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Institute

Mexican Property

Industrial

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PATENT TITLE NO. 342298

Owner (s): XENON PHARMACEUTICALS INC.

Address: 3650 Gilmore Way, V5G 4W8, Burnaby, British Columbia, CANADA

Name: ENANTIOMERS OF SPYROOXINDOL COMPOUNDS AND THEIR USES AS THERAPEUTIC AGENTS.

Classification: lnt.CI.8: A61K31 / 404; A61P29 / 00; C07D491 / 22

Inventors): MIKHAIL CHAFEEV; JIANMIN FU; JEAN-JACQUES CADIEUX

Ι »·

Number:

MX / a'2013 / 008427

Country:

US

Validity: Twenty years

Divil fc &amp;

-I'll

SOUC1TUD.

Feeha for filing June 2010 on Patent Number: 314132

PRIORITY

Date:

June 2009

Number:

61/221,424

Date of Vencühientdft June 28, 2030 .... fe

8L. i&gt;¹ * ·· '' ® reference patent s &amp; torga con-tuidamerttojarii ^ 1st, 2nd section V, 6th section III, and 59 of the Industrial Property Law.

(In accordance with article 23 of the Law of the PrBpMMfmtetnyi, P^resentful The patent is valid for twenty years, which cannot be expired, taken from the date of the loan of the lower status, and will be subject to the payment of the fee to keep people in the stretches.

The person who endorses the present title will establish it with the foundation of the article on articles 6 and 7 and 2 of

Industnal Iopiety (Official Journal # of the Federation (DO R) apaaááfli rsfnn.mda e G2 / 08 / 18M. 25/1/1996, 12/26/1997, 01/01/2004, 06/16/2005, 2 &lt; f01 / 2006,&gt; / 05 / 2009,06 / 01/2010, 2M »X&gt; lÁ U / MOaU yrf &lt;ffÓ4 / 2012); Articles 1,: ¡a a), 4 ° and 12 ° fraccloáps I y lli h Regulations of the institute MSMRHfla'nHBN ^ riaustriai (DOF 14/12/1999,

07/07/2005, 07/07/2004, 2 (007/2004 yíy / 09/2007); articles 1, 3, 4, 5 fraction V inctsoa), 16 fractions I and III and 30 of the Estate MSMHMMMMHIMlPropiedaiMMinamiHeMKIÍ

Law of the 07/05/1999, fraction V formed the

Organic&gt; 07); 1, 3 and 5 Clause a) of the Agreement that delegates powers to the Deputy Directors General, Coordinator, Divisional Directors, Holders of the Regional Offices, Divisional Deputy Directors, Departmental Coordinators and other subordinates of the Mexican Institute of Industrial Property. (DOF 12/15/1999, amended on 02/04/2000, 07/29/2004, 08/04/2004 and 09/13/2007).

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Arenai No. 550, Floor 1.

Coi. Santa María Tepepan town. Xochimiico. CP 16020,

Mexico City

Tei. (55) 53 34 07 00 wwwitnpt ^ ob nix

Issue Date: September 23, 2016

THE DIVISIONAL DIRECTOR OF PATENTS

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NAHANNY CANAL REYES

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MX / 2016/77170 wfee / sl \ 6M.fi ··

IMPI

MEXICAN INSTITUTE OF INDUSTRIAL MONEDAD

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ENANTIOMERS OF SPYROOXINDOL COMPOUNDS AND THEIR USES

AS THERAPEUTIC AGENTS

FIELD OF THE INVENTION

This invention is directed to a specific enantiomer of a spirooxindole compound (spiro-oxindole), specifically to the use of the enantiomer in human or veterinary therapeutics, to treat diseases or conditions in mammals, preferably humans, that are ameliorated or alleviated by modulation, preferably by inhibition of voltage-gated sodium channels.

BACKGROUND OF THE INVENTION

Published PCT Patent Application No. WO 2006/110917, the disclosure of which is incorporated in its entirety by reference herein, discloses certain spirooxindole compounds, in particular T - {[5 (trifluoromethyl) furan-2-yl] methyl} spiro [furo [2,3- / J [1,3] benzodioxol-7,3'-indole] 2 '(1' / - /) - one, in other words, the compound corresponding to the following formula (I) :

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Dt LA TROFISDAD industrial

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These compounds are described in said reference as being useful in the treatment of diseases or conditions, such as pain, in mammals, preferably humans, which are improved or alleviated by modulation, preferably by inhibition, of voltage-regulated sodium channels.

BRIEF DESCRIPTION OF THE INVENTION

The present invention is directed to the discovery that the- (S) enantiomer and- (f?) Enantiomer of the following compound of formula (I):

<img file="MX342298B_D0010.tif" />

demonstrate a difference in potency for inhibition of voltage-gated sodium channel activity.

Accordingly, in one aspect, the invention provides the 1 '- {[5- (trifluoromethyl) furan-2-l] metyl} spiro [furo [2,3Z] enantiomer. [1,3] benzodioxol-7,3'-indole] -2^,(1 '/ - /) - one, that is, the enantiomer- (S) with the

1ΜΠ

MEXICAN INSTITUTE □ t THE INDUSTRIAL PROPERTY

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<img file="MX342298B_D0012.tif" />

(lS) or an acceptable solvate or prodrug thereof. Preferably, the- (S) enantiomer is substantially free of the - (/?) Enantiomer.

In another aspect, the invention provides a pharmaceutical composition comprising the (S) enantiomer, or a pharmaceutically acceptable solvate or prodrug thereof, as stated, preferable and substantially free of the - (/?) Enantiomer, and one or more pharmaceutically acceptable excipients.

In one embodiment, the present invention relates to a pharmaceutical composition comprising the (S) enantiomer, or a pharmaceutically acceptable solvate or prodrug thereof, as stated, preferably and substantially free of the - (/?) Enantiomer, in a pharmaceutically acceptable carrier and in an amount that is effective in treating pain-related diseases or conditions when administered to an animal, preferably a mammal, and more preferably, a human.

In another aspect, the invention provides pharmaceutical therapy in combination with the (S) enantiomer or a pharmaceutically acceptable solvate or prodrug thereof, as stated, preferable and substantially free of the- (ft) enantiomer, and one or more existing therapies. , or

IMPI

INSTITUTO MÍXICANO DI LA PKOPliPAO industrial

<img file="MX342298B_D0013.tif" />

any combination of these, to increase the profile of any existing or future drug therapy, or to decrease adverse events associated with existing or future drug therapy. In one embodiment, the present invention relates to a pharmaceutical composition combining the (S) -enantiomer, or a pharmaceutically acceptable solvate or prodrug thereof, as noted, preferably and substantially free of the- (ft) -enantiomer, with therapies established or future for the indications disclosed in the invention.

In another aspect, the invention provides a method for treating a disease or condition in a mammal, preferably a human, in which the disease or condition is selected from the group encompassing pain, depression, cardiovascular disease, respiratory disease, psychiatric diseases, neurological diseases and seizures, as well as combinations thereof, wherein the method consists of administering to the mammal in need a therapeutically effective amount of the (S) enantiomer, as already stated, or a pharmaceutically acceptable solvate or prodrug thereof.

In another aspect, the invention provides a method of treating pain in a mammal, preferably a human, in which the method comprises administering to the mammal in need a therapeutically effective amount of the (S) -enantiomer, or a solvate or pharmaceutically acceptable prodrug thereof as stated, preferable

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IN!&gt; IISTMal 'and substantially free of the- (R) enantiomer.

In another aspect, the present invention provides a method for treating or alleviating the severity of a disease, condition, or disorder in which the activation or hyperactivity of one or more voltage-regulated sodium channel proteins, which include but not limited to Nav1.1, NaJ.2, Na / I.3, NaJ.4, Na / I.5, Na / I.6, Nav1.7, Na / I.8 voltage regulated sodium channels , or Na / I.9, is involved in the disease, condition, or disorder, method comprising administering to the mammal in need a therapeutically effective amount of the (S) enantlomer, or a pharmaceutically acceptable solvate or prodrug thereof as stated, preferable and substantially free of the (R) enantomer.

In another aspect, the present invention provides a method for the treatment of diseases or conditions in mammals, preferably humans, that are associated with the activity of voltage regulated sodium channels. Accordingly, the invention provides a method for the treatment of diseases or conditions in mammals, preferably humans, that are ameliorated or alleviated by modulation, preferably with inhibition, of voltage-gated sodium channels. Examples of such diseases or conditions include, but are not limited to, pain of any nature and origin, pain associated with HIV, neuropathy induced by HIV treatment, trigeminal neuralgia, post-herpetic neuralgia, diabetic neuropathy, Complex Pain Syndrome Regional (CRPS), Extreme Paroxysmic Pain Disorder (PEPD, en

IMPI

MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY, English), eudinia, heat sensitivity, sarcoidosis, irritable bowel syndrome, Crohns disease, pain associated with multiple sclerosis (MS), motor disability associated with MS, lateral amyotropic sclerosis (ALS, in English), pruritus, hypercholesterolemia, benign prostatic hyperplasia, peripheral neuropathy, arthritis, rheumatoid arthritis, osteoarthritis, paroxysmic dystonia, periodic paralysis, myasthenia syndromes, myotonia, malignant hyperthermia, cystic fibrosis, pseudoaldosteronism, rhabdomyolysis, bipolar depression, anxiety, schizophrenia, disease due to exposure to insecticides or other agents that promote neuronal or muscular hyperexcitability, hereditary erythemagia, hereditary rectal pain, hereditary facial pain, migraine, headache, neuralgiform headache, hereditary hemiplegic migraine, conditions associated with headache, sinus pain, tension headache, phantom limb pain, peripheral nerve damage, cancer, epilepsy, general and partial tonic stoppages, restless leg syndrome, arrhythmias, fibromyalgia, neuroprotection under ischemic conditions caused by an attack, glaucoma or neural trauma, tachyarrhythmias , atrial fibrillation and ventricular fibrillation, in which the method comprises, administering to the mammal in need a therapeutically effective amount of the- (S) enantiomer, or a pharmaceutically acceptable solvate or prodrug thereof, as stated, preferably and substantially free of the (R) enantiomer.

In another aspect, the invention provides a method for treating a disease or condition in a mammal, preferably

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IMPI

MEXICAN INSTITUTE

OF THE PROPERTY

INDUSTRIAL a human, by inhibiting ion flow through a voltage-gated sodium channel in the mammal, in which the method comprises administering to the mammal in need a therapeutically effective amount of the (S) -enantiomer, or a solvate or a pharmaceutically acceptable prodrug thereof, as already stated, preferably and substantially free of the - (/?) enantiomer.

The invention further provides for the use of the (S) enantiomer, or a pharmaceutically acceptable solvate or prodrug thereof, as noted, preferable and substantially free of the (R) enantiomer, in the preparation of a medicinal composition for the treatment of a disease or condition associated with the activity of a voltage-regulated sodium channel. Accordingly, the invention provides for the use of the (S) enantiomer, or a pharmaceutically acceptable solvate or prodrug thereof, as stated, preferably and substantially free of the (R) enantiomer, in the preparation of a medicinal composition for the treatment of a disease or condition that is ameliorated or alleviated by modulation, preferably with the inhibition, of a voltage regulated sodium channel.

BRIEF DESCRIPTION OF THE DRAWINGS

The following drawings are part of the present specification and are included to demonstrate in more detail some aspects of the present invention. The invention can be better understood by doing

IMPI

Mexican Institute of Industrial Property

<img file="MX342298B_D0016.tif" />

Reference to one or more of the drawings in combination with the detailed description of specific proposals presented here.

FIG. 1 presents the concentration-response relationship for the - (S) and - (R) enantiomers in the Guanidine Incoming Flow Assay of Biological Example 1.

FIG. 2 shows a comparison of the efficacy of the - (S) and - (/?) Enantiomers with an oral dose in an inflammatory pain model of Biological Example 3.

FIG. 3 shows a comparison of the efficacy of - (S) and - (/?) Enantiomers with topical administration in a neuropathic pain model of Biological Example 3.

FIG. 4 shows the progress in histamino-induced itching time in untreated mice in the in vivo assay described in Biological Example 7. Data are expressed as Mean ± SD (standard deviation) of the itching periods.

FIG. 5 shows the efficacy against a histaminoinduced itch of an ointment with a content of 8% (w / v) of the enantiomer- (S), applied topically. Data are expressed as Mean ± SD (standard deviation) of the itching periods.

FIG. 6 shows the efficacy of the (S) enantiomer against histamino-induced itching when administered orally rather than topically. Data are expressed as Mean ± SD (standard deviation) of the itching periods.

IMPI

MEXICAN INSTITUTE OE THE PROPERTY

INDUSTRIAL

DETAILED DESCRIPTION OF THE INVENTION

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Definitions

As used in the specification and in the appended claims, 5 unless otherwise specified, the following terms have the indicated meaning:

"Analgesia" refers to an absence of pain in response to a stimulus that would normally be painful.

"Alodynia" refers to a condition in which a normally harmless sensation, such as pressure or a light touch, is perceived as painful.

"Enantiomers refers to asymmetric molecules that can exist in two different isomeric forms whose spatial configurations are different. Other terms used to designate or refer to enantiomers include "stereoisomers" (due to different arrangement or stereochemistry around the chiral center; although all enantiomers are stereoisomers, not all stereoisomers are enantiomers) or "optical isomers" (due to optical activity of pure enantiomers, which consists of the ability of different pure enantiomers to rotate plane polarized light in different directions). Because they do not have a plane of symmetry, enantiomers are not identical to their mirror images; molecules that exist in two enantiomeric forms are chiral, which means that they can be considered to exist in forms

IMPI

MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY

<img file="MX342298B_D0018.tif" />

"left and right. The most common cause of chirality in organic molecules is the presence of a tetrahedral carbon attached to four different substitutes or groups. Such carbon is referred to as the chiral center or stereogenic center. One method of indicating the three-dimensional arrangement of atoms 5 (or configuration) in a stereogenic center is to refer to the arrangement of the priority of the groups when the group of least priority is oriented opposite the hypothetical observer: if the arrangement of the three remaining groups, from highest to lowest priority, is clockwise (clockwise), the stereogenic center has a “K (or“ D ”) configuration; if the arrangement is in the opposite direction, the stereogenic center has an "S" (or "L") configuration.

Enantiomers have the same empirical formula and are generally chemically identical in their reactions, their physical properties, and their spectroscopic properties. However, enantiomers show different chemical reactivity towards other asymmetric compounds, and respond differently to asymmetric physical alterations. The most common asymmetric alteration is polarized light.

An enantiomer can rotate polarized light; this indicates that an enantiomer is optically active. Two different enantiomers of the same compound will rotate the polarized light in opposite directions; thus, the light can be rotated counterclockwise or counterclockwise for a hypothetical observer (this is called a levorotatory or "I, or" minus, "or" - "), or it can be rotated counterclockwise. to the right, or clockwise, (to

IMPI • Mexican MSTITinv BS INDUSTRIAL PROPERTY

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this is called a dextrorotatory or "d, or" plus, "or The sign of optical rotation (+) or (-) is not related to the R, S designation. A mixture of equal amounts of two chiral enantiomers is It is called a racemic mixture or racemate and is designated by one of the two signs (+/-) or by the prefixes "d, l" ​​to indicate a mixture of left and right ways. The compound of formula (I) described here is a racemic mixture. Racemic mixtures show zero optical rotation because equal amounts of (+) and (-) forms are present. In general, the presence of a single enantiomer rotates polarized light in only one direction; therefore, an enantiomer is only referred to as optically pure.

The "R and" S designations are used to indicate the absolute configuration of a molecule around its chiral center (s). The designation may appear as a prefix or as a suffix; they may or may not be separated from the enantiomer name by a hyphen; they may or may not be scripted; and they may or may not be enclosed in parentheses.

The designations or prefixes "(+) and (-)" are used here to designate the compound-induced polarized light rotation sign, the () indicating that the compound is left-handed (rotated counterclockwise). The compound with the prefix (+) is clockwise (rotates clockwise).

"Resolution" or "solving: when these terms are used in reference to a racemic mixture or racemic compound they refer to the separation of the racemic mixture into its two enantiomeric forms, (ie, its (+) and (-) forms; (R) and (S).

IMPI

MEXICAN INSTITUTE OF Ι.Λ industrial PROPERTY

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"Enantiomeric excess" or "ee refers to-ae-pfeducia-eu-elxLLal one enantiomer is present in excess of the other, and is defined as the absolute difference in mole fraction between both enantiomers. The enantiomeric excess is typically expressed as the percentage of a 5 enantiomer present in the mixture relative to the other enantiomer. For the purposes of this invention, the (S) enantiomer is considered to be "substantially free" of the (R) enantiomer when the- (S) enantiomer is present in enantiomeric excess greater than 80%, preferably greater than 90%, more preferably greater than 95%, and most preferably when greater than 99%.

The chemical naming protocol and structural diagrams used here are a modified form of the IUPAC naming system {International Union of Puré and Applied Chemistry¹) applied through the ACD / Name Version 9.07 program. For example, the compound of formula (I), as indicated above in the Summary of the Invention, is here called r - {[5- (trlfluoromethyl) furan-2yl] methyl} spiro [furo [2, 3 - /] [1,3] benzodioxol-7,3'-indole] -2 '(1' / - /) - one. The corresponding (S) enantlomer, that is, the- (S) enantlomer of the formula (lS), as indicated above in the Summary of the Invention, is here called (S) -1 '- {[520 (ϋ · ΙΑυοΓθΐτιβΙΙΙ) ίυΓ3η-2-ΙΙ] πιβύΙ} 8ρΐΓθ [ίυΓθ [2,3 - /] [1,3] όβηζοόΙοχοΙ-7,3'-ΙηόοΙ] 2 '(1' / - /) - one. The corresponding (R) enantlomer, the- (R) enantiomer of the following formula (I- /?):

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or a pharmaceutically acceptable solvate or prodrug thereof, is referred to herein (/?) - T - {[5- (tr¡fluoromet¡l) furan-2¡l] met¡l} sp¡ro [furo [2, 3 - /] [1,3] benzodioxol-7.3^,-¡Ndol] -2^,(1Y7) -ona.

"Prodrug is used to indicate a compound that can be converted under physiological conditions or by solvallsis into a biologically active compound of the invention. Thus, the term "prodrug" refers to a metabolic precursor of a compound of the invention that is pharmaceutically acceptable. A prodrug can be inactive when administered to a patient in need, but is converted in vivo to an active compound of the invention. Typically, prodrugs are rapidly transformed, in vivo, to generate a parent compound of the invention, for example by hydrolysis in blood. The prodrug compound often offers solubility, tissue compatibility, or delayed release benefits in a mammalian organism (see, Bundgard, H., Design of Prodrugs (1985), pp. 7-9, 21-24 (Elsevier, Amsterdam)). In Higuchi, T., et al., Pro-drugs as Novel Delivery Systems, ACS Symposium Series, Vol. 14, and in Bioreversible Carriers in Drug Design, Ed. Edward B. Roche, American Pharmaceutical Association and Pergamon Press, 1987, a discussion on

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prodrugs. Both works are incorporated in their entirety by reference in the present memory.

The term "prodrug" is also intended to include any covalently linked carrier that releases the active compound of the invention in vivo when said prodrug is administered to a mammalian subject. Prodrugs of a compound of the invention can be prepared by modifying the functional groups present in the compound of the invention in such a way that the modifications are separated (from ved), either by routine manipulation or in vivo, to the parent compound of the invention.

Prodrugs include compounds of the invention in which a hydroxyl, amino or mercaptan group is linked to any group that when the prodrug of the compound of the invention is administered to a mammalian subject, separates to form a free hydroxyl group, a free amino group or a free mercaptan group, respectively. Examples of prodrugs include, but are not limited to, derivatives of alcohol acetates, formats and benzoates or amide derivatives of amino functional groups present in the compounds of the invention and the like.

The invention disclosed herein also contemplates encompassing the (S) enantiomer and the (fR-enantiomer) disclosed herein, isotopically labeled with having one or more atoms replaced by an atom with a different atomic mass or atomic number. Examples of isotopes that can be incorporated into the compounds disclosed herein include the isotopes of hydrogen, carbon, nitrogen, oxygen, such as²H ³H ¹¹C, ¹³C, ¹⁴C, ¹³N,

IMPI

Mexican Institute of Industrial Property

<img file="MX342298B_D0023.tif" />

¹⁵N, ¹⁵OR, ¹⁷OR, ¹⁸O, and ¹⁸F, respectively. —These radiolabelled compounds could be useful in helping to determine or measure the efficacy of compounds, characterizing, for example, the site or mode of action on voltage-gated sodium channels, or the binding affinity to a site pharmacologically important action on voltage-regulated sodium channels. The isotopically identified compounds are useful in drug and distribution studies on tissue substrates. Radioactive isotopes of tritium, i.e. ³H, and carbon-14, that is, ¹⁴C, are particularly useful for this purpose given their ease of incorporation and availability of detection means. A radioligand incorporating tritium (³H) is particularly useful in studies of ligand linkages with membranes containing voltage-regulated sodium channels because tritium has a long half-life of decomposition and the emission is of relatively low energy, and therefore the radioisotope is relatively safe .

Typically, the radioligand is prepared by exchanging tritium with hydrogen in an unlabelled compound. Identification of active and non-active enantiomers of a particular racemic mixture facilitates the development of a ligand binding assay because the unlabeled inactive enantiomer can be added to the assay to somehow reduce, eliminate or control non-specific binding of the active tritiated enantiomer .

Substitution with heavier isotopes such as deuterium, i.e. ²H, can mean certain therapeutic advantages resulting from greater metabolic stability, such as, for example, an increased half-life in

IMPIOS

MEXICAN INSTITUTE D £ THE PROPERTY

INDUSTRIAL live or smaller dose requirements, and may therefore be preferred in some circumstances.

Substitution with positron-emitting isotopes, such as ¹¹C, ¹⁸F, ¹⁵0 and ¹³N, may be useful in Positron Emission Topography (PET) studies to examine occupancy in the recipient substrate. Isotopically-labeled enantlomers of the invention can generally be prepared by conventional techniques known to those skilled in the art or by processes analogous to those described herein using suitable, isotopically-labeled reagents in place of the previously unlabelled reagent.

The invention disclosed herein also contemplates encompassing the in vivo metabolic products generated by the disclosed enantlomers. Such products may be the result of, for example, oxidation, reduction, hydrolysis, amplification, esterification, and the like, of the administered compound, primarily due to enzymatic processes. Accordingly, the invention includes metabolic products generated by a process comprising contacting an enantiomer of the present invention with a mammal, for a period of time sufficient for the metabolic product to be generated. Such metabolic products can be

Identified by administering to an animal, such as a rat, a mouse, a guinea pig or acure (guinea pig), a monkey, or a human, a radiolabelled enantiomer of the invention, in a detectable dose and allowing sufficient time to metabolism is complete, and then

INSTITUTE MBXiCANf OF INDUSTRIAL PROPERTY isolating the metabolic product from urine, blood or other biological samples.

"Selectivity" and "selective" are used herein as a relative measure of the tendency of a compound of the invention to preferentially associate with one thing and not with another (or a group of others), such as between sodium channels regulated by voltage. For example, selectivity can be determined by comparative measurements of kinetics and affinity for balanced binding and / or by functional measurements of ion transport through voltage regulated sodium channels. The tendency of a compound to associate with a voltage regulated sodium channel can be measured by different techniques, and many types of association are known to those skilled in the art, as disclosed in other sections of the present specification. Selectivity means that in a particular type of association, measured in a specific way, a compound exhibits a tendency or preference to associate with one voltage-regulated sodium channel rather than one or more other voltage-regulated sodium channels. This association may be different for different types of tests or different ways of measuring.

"Stable enantiomer" and "stable structure are intended to indicate a compound robust enough to survive isolation to a useful degree of purity from a reaction mixture, and then its

I formulation to an effective therapeutic agent.

"Mammal includes both humans and pets,

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MEXICAN INST1TUT OF THE INDUSTRIAL PROPERTY

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laboratory animals and domestic pets (eg, cats, dogs, pigs, cattle, sheep, goats, horses, and rabbits) as well as non-domestic animals such as wildlife and the like.

"Pharmaceutically acceptable carrier, diluent or excipient" includes without limitation any adjuvant, carrier, excipient, "glidant", sweetening agent, diluent, preservative, dye / flavor enhancer, surfactant, wetting agent, dispersing agent, agent suspension, stabilizer, isotonic agent, solvent or emulsifier that has been approved for use in humans and pets by, as a non-limiting example, the United States Food and Drug Administration (FDA), Health Canada (Health Canada) or the European Medicines Agency (European Medicines Agency).

"Pharmaceutical composition" refers to a formulation of a compound of the invention and a generally art-acceptable medium for the delivery of the biologically active compound to mammals, eg, humans. Such a medium includes all of the aforementioned pharmaceutically acceptable carriers, diluents or excipients.

The pharmaceutical compositions of the invention comprise one or more pharmaceutically acceptable excipients, including but not limited to, any solvent, adjuvant, bioavailability enhancer, carrier, glidant, sweetening agent, diluent, preservative, dye / colorant. , flavor enhancer, surfactant, agent

<img file="MX342298B_D0025.tif" />

wetting agent, dispersing agent, self-suspending agent, ..... stabilizing agent, isotonic agent, chemical buffer (ou / fer) and / or emulsifier that has been approved for use in humans and pets by, as a non-limiting example, the United States Food and Drug Administration (FDA), Health Canada (Health Canada) or the European Medicines Agency (European Medicines Agency). By way of example, pharmaceutically acceptable excipients include but are not limited to the following:

benzyl alcohol benzyl benzoate caprylocaproyl macrogolglycerides {eg Labrasol®) dimethyl amine (DMA) ethanol

2- (2-ethoxyethoxy) ethanol {eg Transcutol®) glucose (solution) glyceryl caprylate / caprate complex and PEG-8 (polyethylene glycol) caprylate / caprate (eg ,, Labrasol®) isopropyl alcohol

Lauroyl Macrogol-32 Glycerides {eg Gelucire® 44/14) macrogol-15 hydroxystearate {eg, Solutol® HS15) medium chain triglycerides {eg, Miglyol® 810, Miglyol® 840 or

Miglyol® 812) peanut oil

IMPI

MEXICAN INSTITUTE DF. INDUSTRIAL ROPISIS polysorbate 80 (eg Tween® 80) polyethylene glycol (PEG) polyethylene glycol 400 (PEG400, eg Lutrol® E 400) polyethylene glycol 6000 polyoxyl 35 castor oil (eg Cremophor® EL) polyoxyl 40 hydrogenated castor oil (eg ,, Cremophor® RH

40) propylene glycol (PG) propylene glycol monocaprylate (Capryol® 90) soybean oil sulfobutylether-p-cyclodextrin (eg, Capitsol®)

TPGS (α-tocopherol polyethylene glycol succinate) water

Other pharmaceutically acceptable excipients are disclosed herein.

It often happens that crystallizations produce solvates of the compound of the invention. In use herein, the term "solvate" refers to an aggregate comprising one or more molecules of a compound of the invention with one or more molecules of the solvent. The solvent can be water, in which case the solvate can be a hydrate. Alternatively, the solvent can be an organic solvent. Accordingly, the compounds of the present invention may exist as hydrates, including a

<img file="MX342298B_D0026.tif" />

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MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY

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monohydrate, dihydrate, hemihydrate, sesquihydrate, trihydrate, tetrahydrate and the like, as well as the corresponding solvated forms. The compound of the invention may be true solvates, while in other cases the compound of the invention may only retain adventitious water or be a mixture of water plus some adventitious solvent.

"Therapeutically sufficient amount" refers to that amount of a compound of the invention that when administered to a mammal, preferably a human, is sufficient to serve as a treatment, as defined below, of a disease or condition of interest in the mammal, preferably a human. The amount of a compound of the invention that constitutes a "therapeutically sufficient amount" will vary depending on the compound, the condition and its severity, the form of administration and the age of the treated mammal, but may be routinely determined by someone with ordinary skill in art that applies its own knowledge and this disclosure.

"Treatment or" treatment as used herein covers the treatment of a disease or condition of a mammal, preferably a human, suffering from the disease or condition of interest, including:

(i) prevent the disease or condition from occurring in the mammal, particularly when that mammal is predisposed to the condition but has not yet been diagnosed;

(ii) inhibit the disease or condition, that is, stop (arrest) its development;

IMPIOS

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INDUSTRIAL (ii) alleviate the disease or condition, that is, cause regression of the disease or condition;

(iv) alleviate the symptoms resulting from the disease or condition, that is, alleviate pain with or without attending to the causing disease or condition.

As used herein, the terms "improving", "improved", "relieving" or "relieved" are to be understood according to their generally accepted definitions. For example, "improve" generally means making a condition better relative to the same condition that prevailed before the improving event. "Relieving" generally means making a condition more bearable relative to the same condition that prevailed before the relief event. As used herein, "improving" or "improved" may refer to a disease or condition that is best done by administration of a compound of the invention. As used herein, "alleviating" or "alleviated" may refer to a disease or condition that is made more bearable by the administration of a compound of the invention. For example, "relieving" pain would include reducing the severity or amount of pain.

As used herein, "disease" and "condition may be used interchangeably or may differ in that the particular disease or condition may not have a known causative agent (ie the etiology has not been completed) and therefore so much can still not be recognized as a disease but only as a condition

IMPI

INSTITUTO MEXICANO D £ THE INDUSTRIAL PROPERTY undesirable or syndrome, in which a set of more or less specific symptoms have been identified by clinical professionals.

Utility and tests of the compounds of the invention

The present invention relates to the (S) -enantiomer of T {[5- (trifluoromethyl) furan-2-yl] methyl} spiro [furo [2,3 - /] [1,3] benzodioxol-7,3 ' -indole] 2 '(T / - /) - one, with pharmaceutical compositions and methods making use of the (S) -enantiomer of the invention and pharmaceutical compositions for the treatment of diseases or conditions that are ameliorated or alleviated by modulation, preferably inhibition of voltage regulated sodium channels; preferably diseases and conditions related to pain and itching; central nervous system conditions such as epilepsy, restless leg syndrome, anxiety, depression, and bipolar disease; cardiovascular conditions such as arrhythmias, atrial fibrillation, and ventricular fibrillation; neuromuscular conditions such as muscle paralysis, myotonia, or tetanus; neuroprotection against attacks, neural trauma and multiple sclerosis; and channelopathies such as erythromelalgia and hereditary rectal pain syndrome, by administering to a patient in need of such treatment an effective amount of a voltage-gated sodium channel blocking agent, which modulates, and especially inhibits, and which preferably it comes from the enantiomers of the invention.

In general, the present invention provides a method for

IMPI

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<img file="MX342298B_D0028.tif" />

treatment of a mammal, preferably a human, suffering from, or to prevent a mammal, preferably a human, from developing a disease or condition associated with the activity of voltage-gated sodium channels, especially pain, in which the method comprises administration to the mammal of a therapeutically effective amount of the (S) -enantiomer, or of a pharmaceutically acceptable solvate or prodrug thereof, as set forth in the Summary of the Invention, in which the (S) enantiomer modulates, preferably inhibits, the activity of one or more voltage regulated sodium channels.

The family of voltage regulated sodium channel proteins has been extensively studied and as a result their involvement in a number of vital bodily functions has been demonstrated. Research in this area has identified variants of the alpha subunits that result in major changes in channel functions and activities, changes that may eventually lead to major pathophysiological conditions. Additionally, excessive sodium intake can arise indirectly via inflammatory agents or factors that result in hyperexcitability. Implicitly with function, this family of proteins is considered a point of first choice for therapeutic intervention. The voltage-gated sodium channel proteins Nay1.1 and Nay1.2 have high expression in the brain (Raymond, CK, et al., J. Biol. Chem. (2004), 279 (44): 46234-41) and are vital for normal brain function. In humans, mutations of Na_v1.1 and Na / I.2 result in epileptic states and in

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MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY some cases of mental decline and migraines (Rhodes, TH, et ai, Proc. Nati.

<img file="MX342298B_D0029.tif" />

Acad. Sci. USA (2004), 101 (30): 11147-52; Kamiya, K., et al., J. Blol. Chem. (2004), 24 (11): 2690-8; Pereira, S. et al., Neurology (2004), 63 (1): 191-2; Meisler, MH et al., J. Physiol. (Lond.) (In press). Portal motifs, both channels have been considered validated targets for the treatment of epilepsy (see PCT Published Patent No. WO 01/38564).

Nay1.3 is primarily expressed in the central nervous system of neonatal animals, and at low levels throughout the adult body (Raymond, CK, et al., Op. Cit.). After damage to the nervous system, its expression has been shown to be upregulated in rat dorsal horn neurons (Hains, BD, et al., J. Neuroscl. (2003), 23 (26): 8881-92). Na / I.3 has been considered by many experts in the field to be a suitable target for pain therapy because its expression is induced by nerve damage (Lai, J., et al., Curr. Opin. Neurobiol. (2003) , (3): 291-72003: Wood, JN, et al., J. Neurobiol. (2004), 61 (1): 55-71: Chung,

JM, et al., Novartis Found Symp. (2004), 261: 19-27; discussion 27-31,47-54; Priest, BT, Curr. Opin. Drug Discov. Devel. (2009) 12: 682-693).

The expression of Na / I.4 is essentially limited to the muscles (Raymond, CK, et al., Op. Cit.). Mutations of this gene have been shown to have profound effects on muscle function, including paralysis (Tamaoka A., Intem. Med. (2003), (9): 769-70). For this reason, this channel is considered a target for the treatment of periodic paralysis, myotonia, abnormal muscle contractibility, spasms or paralysis.

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The voltage-gated cardiac sodium channel ^ Wai / I ^ T'Sé is expressed primarily in cardiac myocytes (Raymond, CK, et al., Op. Cit.), And is found in the atria, ventricles, the sinoatrial node, the atrioventricular node and Purkinge cells. The rapid upward stroke of cardiac action and the rapid conduction of the impulse through cardiac tissue are due to the opening of Na / 1.5. As such, Na / 1.5 is linked to cardiac arrhythmias. Mutations of human Na / 1.5 result in multiple arrhythmic syndromes, including, for example, long QT3 (LQT3), Brugada syndrome (BS), an inherited defect in cardiac conduction, death syndrome Sudden Unexpected Nocturnal (SUNDS) Sudden Infant Death Syndrome (SIDS) (Liu, H. et al., Am. J. Pharmacogenomics (2003), 3 (3): 173-9; Ruan, Y et al., Nat. Rev. Cardiol. (2009) 6: 337-48). Voltage-regulated sodium channel blocking therapy has been widely used in the treatment of cardiac arrhythmias. The first antiarrhythmic drug, quinidine, discovered in 1914, is classified as a sodium channel blocker.

Na / I.6 encodes an abundant, voltage-regulated sodium channel widely distributed in the central and peripheral nervous systems, clustered in the Ranvier nodes of the neural axons (Caldwell, JH, et al., Proc. Nati. Acad. Sci. USA (2000), 97 (10). 5616-20). As a result of mutations leading to loss of function in mice, ataxia and seizures have been detected (Papale, LA et al., Human Mol. Genetics (2009) 18, 1633-1641). Although they have not been detected

IMPI

INSTITUTO MEXICANO HE LA MONEDAD INDUSTRIAL mutations in humans, it is believed that Na / 1.6 plays a role in

<img file="MX342298B_D0030.tif" />

manifestation of symptoms associated with multiple sclerosis and therefore it has been considered a target for the treatment of this disease (Craner, MJ, et al., Proc. Nati. Acad. Sci. USA (2004), 101 (21): 8168-73).

Na / I.7 is primarily expressed in the peripheral nervous system, both in sensory and sympathetic neurons (Raymond, CK, et al., Op. Cit.). Mutations that lead to loss of function in humans cause congenital pain indifference (CIP) without impairing cognitive or motor functions (Cox, JJ et al., Nature (2006)

444 (7121), 894-8; Goldberg, YP et al., Clin. Genet. (2007) 71 (4), 311-9).

Individuals with CIP do not experience inflammatory or neuropathic pain, suggesting that the selective Nat / I.7 block would eliminate multiple forms of chronic and acute pain without a damaging effect on the central or peripheral nervous systems, or on a muscle. Furthermore, the single nucleotide polymorphism (R1150W), which has very subtle effects on time and voltage dependence of the Na / I.7 gate action, has great effects on pain perception (Reimann, F. et al., Proc. Nati. Acad. Sci. USA (2010), 107 (11), 5148-53). About 10% of patients with a variety of pain conditions are heterozygous for the allele that confers greater pain sensitivity. The involvement of Na / I.7 in mediating pain responses is also evidenced by an increase in mutations resulting in erythromelalgia or extreme paroxysmal pain disorder (DibHajj SD et al., Adv. Genet. (2009) 63 : 85-110). Although Na / I.7 is

IMPI

<img file="MX342298B_D0031.tif" />

MEXICAN INSTITUTE

Give THE ERORIETY V * · INDUSTRIAL V, expressed mainly in the peripheral nervous system, a point mutation of Nay1.7 causes febrile seizures, indicating that this channel plays a role in the central nervous system (CNS). Thus, voltage-regulated sodium channel blockers can be useful as anticonvulsant agents.

Na / 1.8 expression is predominant in the dorsal root ganglia (DRG) (Raymond, CK, et al., Op. Ct.). The elevation of the action potential of the DRG sensory neurons is transported mainly by current through Nay1.8, so that blocking this current will possibly block pain responses (Blair, NT and Bean, BP, J. Neurosd. 22: 10277-90). Consistent with this finding, the deactivation (knock-down) of Nav1.8 in rats has been achieved using antisense (antisense) DNA or small interfering RNAs, obtaining virtually complete reversal of neuropathic pain in models of ligation pain and chronic constriction of the spinal nerve. A selective Nay1.8 blocker has been reported and is effective in blocking both neuropathic and inflammatory pain (Jarvis, MF et al., Proc. Nati. Acad. Sd. USA (2007), 104 (20), 8520-5). PC Patent T Published No.

WO03 / 037274A2 describes pyrasolamides and sulfonamides for the treatment of central or peripheral nervous system conditions, particularly pain and chronic pain, by blocking sodium channels associated with the onset (recurrence) of the indicated conditions. PCT Published Patent No. W003 / 037890A2 describes piperidines for the treatment of

<img file="MX342298B_D0032.tif" />

central or peripheral nervous system conditions, particularly pain and chronic pain, by blocking sodium channels associated with the onset or recurrence of the indicated conditions. The compounds, compositions, and methods of these inventions are of particular use for the treatment of neuropathic or inflammatory pain by inhibiting the flow of ions through a channel that includes a PN3 subunit (Na / I.8).

The Na / I.9 voltage-regulated sodium channel of the peripheral nervous system, reported in Dib-Hajj, SD, et al. (see Dib-Hajj, SD, et al., Proc. Nati. Acad. Sel. USA (1998), 95 (15): 8963-8), expressed in the dorsal root ganglia. Na / I.9 has been shown to underlie neurotropin-activated depolarization and excitation (BDNF). The limited expression pattern of this channel has made it a candidate for pain management (Lai, J, et al., Op. cit .; Wood, JN, et al., Op. cit .; Chung, JM et al., Op. cit.).

NaX is a putative sodium channel for which it has not yet been shown to be voltage regulated. In addition to its expression in the lungs, heart, dorsal root ganglia, and Schwann cells of the peripheral nervous system, NaX is found in neurons and ependymal cells (ependymaf) in restricted areas of the CNS, particularly in the circumventricular organs, which participate in body fluid homeostasis (Watanabe, E., et al., J. Neurosci. (2000), 20 (20): 7743-51). Mice without NaX (NaX-null) showed abnormal inputs of hypertonic saline under both low water and low salt conditions. These

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INDUSTRIAL

<img file="MX342298B_D0033.tif" />

Findings suggest that NaX plays an important role in the central sensing of sodium level in body fluids and in the regulation of salt absorption behavior. Its pattern of expression and function suggest it as a target for the treatment of cystic fibrosis and other ailments related to salt regulation.

Studies with tetrodoxin (TTX), a voltage-regulated sodium channel blocker used to reduce neuronal activity in certain regions of the brain, point to its potential use in the treatment of addiction. Drug paired stimuli elicit cravings and relapses in addicted rats and drug-seeking behaviors in rats. The functional integrity of the bajolateral amygdala (basolateral amygdala -BLA) is necessary to restore the cocaine-seeking behavior caused by stimuli conditioned by cocaine, but not by cocaine itself. The BLA plays a similar role in the restitution of heroin-seeking behaviors. TTX induced BLA deactivation in heroin-conditioned, conditioned restitution of extinct heroin-seeking behavior in a rat model Fuchs, RA and See, RE, Psychopharmacology (2002) 160 (4): 425-33).

A subset of C fibers mediate responses to pruritogenic agents, especially histamine itching, PAR-2 ​​receptor activators, cholestasis, and viral infections Steinhoff, M. et al., J. Neurosci. 23: 6176-80; Twycross, R. et al., QJ Med. 96: 7-26). Voltage regulated sodium channels are expressed in and mediated in the impulses of

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MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY C-fiber nerves (C-fiber). ____

The overall value of the (S) -enantiomer of the invention for modulating, especially inhibiting, ion flow through voltage regulated sodium channels can be determined from the assays described below in the Biological Assays section. Alternatively, the general value of the (S) enantiomer of the invention for the treatment of conditions and diseases can be established by animal models that are industry standard to demonstrate the efficacy of pain treatment compounds. Animal models of neuropathic pain conditions in humans that result in reproducible sensory deficits (allodynia, hyperalgesia, and spontaneous pain) over a sustained period of time have been developed that can be evaluated by sensory testing. By establishing the levels of allodynia and hyperalgesia present, mechanically or chemically induced, and by temperature, various pathophysiological conditions observed in humans can be modeled, allowing the evaluation of drug therapies.

In rat peripheral nerve injury models, ectopic activity on the injured nerve correlates with behavioral signs of pain. In these models, the application of the (S) -enantiomer of the invention and local anesthetic lidocaine in concentrations that do not affect general behavior and motor function can suppress ectopic activity and reverse tactile allodynia (Mao, J. and Chen, LL, Pain (2000), 87: 717). The alimetric scaling of the effective doses in these models with

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INDUSTRIAL **: ---- 10 rats results in doses similar to those that have been shown to be effective in humans (Tanellan, DL and Brose, WG, Anesthesiology (1991), 74 (5): 949-951). Furthermore, Lldoderm®, lldocaine applied in the form of a dermal patch, is an FDA-approved treatment of postherpetic neuralgia (Devers, A. and Glaler, BS, Clin. J. Pain (2000), 16 (3 ): 205-8).

Voltage-regulated sodium channel blockers have additional clinical uses for pain. Recent evidence from animal models suggests that voltage-gated sodium channel blockers may also be useful for neuroprotection under ischemic conditions caused by seizures or neural trauma, and in patients with multiple sclerosis (MS) (Clare, JJ et al ., op. cit. and Anger, T. et al., op. cit.).

The (S) -enantiomer of the invention modulates, preferably inhibits, ion flow through a voltage regulated sodium channel in a mammal, especially a human. Any modulation, whether partial or complete inhibition, or prevention of ionic flow, is referred to herein as "blocking" and the corresponding compounds as "blockers" or "inhibitors. In general, the compound of the invention modulates the activity of a voltage-gated sodium channel down, inhibits the voltage-dependent activity of the voltage-gated sodium channel, and / or reduces or prevents sodium ion flow through a cell membrane preventing the activity of the sodium channel regulated by voltage such as ionic flow.

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INSTITUTO MEXCANO 0Γ LA PKOHEDAD ¡NDUSTMAL

<img file="MX342298B_D0034.tif" />

The (S) -enantiomer of the invention is a sodium channel blocker and is therefore useful in the treatment of diseases and conditions in mammals, preferably humans, and other organisms, including all those diseases and conditions that are the result of the aberrant activity of voltage-gated sodium channels or that can be enhanced or alleviated by modulation, preferably inhibition, of the biological activity of voltage regulated sodium channels.

As defined herein, a disease or condition that is ameliorated or alleviated by modulation, preferably inhibition, of a voltage regulated sodium channel, refers to a disease or condition that is ameliorated or ameliorated by modulation, preferably inhibition of the voltage-gated sodium channel, and includes, but is not limited to, pain and itching; central nervous system conditions such as epilepsy, anxiety, depression (Morinville et al., J. Comp.

Neurol., 504: 680-689 (2007)) and bipolar disease (Ettinger and Argoff, Neurotherapeutics, 4: 75-83 (2007)); cardiovascular conditions such as arrhythmias, atrial fibrillation, and ventricular fibrillation, neuromuscular conditions such as restless leg syndrome, and muscle or tetanus paralysis; neuroprotection against attacks, neural trauma and multiple sclerosis; and channelopathies such as erythromelalgia and hereditary rectal pain syndrome.

Additional diseases and conditions include pain associated with HIV, neuropathies induced by HIV treatment, neuralgia

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<img file="MX342298B_D0035.tif" />

trigeminal, glossopharyngeal neuralgia, neuropathy secondary to metastatic irrigation, painful adiposis, thalamic lesions, hypertension, autoimmune disease, asthma, drug addiction (eg, opiates, benzodiazepine, amphetamine, cocaine, alcohol, butane inhalation), Alzheimer's disease (Kim DY, Carey et al., Nat. Cell Biol. 9 (7): 755-764 (2007)), dementia, age-related memory disability, Korsakoff syndrome, restenosis, urinary dysfunction, incontinence, Parkinson's disease (Do and Bean, Neuron 39: 109-120 ( 2003); Puopolo et al., J. Neurosci. 27: 645-656 (2007)), cerebrovascular ischemia, neurosis, gastrointestinal disease, sickle cell anemia, heart failure, myocardial infarction, reperfusion injury (reperfusion), intermittent claudication , angina, seizures, respiratory disorders, cerebral or myocardial ischemia, long-QT syndrome, polymorphic catecholeminergic ventricular tachycardia, ophthalmic diseases, spasticity, Spastic parapiegia, myopathies, myasthenia gravis, paramyotonia congenita, periodic hyperkalemic paralysis, periodic hypokalemic paralysis, alopecia, anxiety disorders, psychotic disorders, mania, paranoia, seasonal affective disorder, panic disorder, obsessive-compulsive disorder (OCD, English) , phobias, autism, Aspergers syndrome, Retts syndrome, disintegrative disorder, attention deficit disorder, aggressiveness, impulse control disorders, thrombosis, pre-eclampsia, congestive failure, cardiac arrest, Freidrich's ataxia, spinocereberal ataxia, tremor, weakness

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<img file="MX342298B_D0036.tif" />

muscle, myelopathy, radiculopathy, systemic lupus erltomatosus, granulomatous disease, olive-ponto-cerebellar atrophy, spinocereberal ataxia, episodic ataxia, myochemy, progressive pallid atrophy, progressive supranuclear palsy and traumatic brain damage, hydroedema the spine, anorexia nervosa, bulimia, Prader-Willi syndrome, obesity, optic neuritis, cataracts, retinal hemorrhage, ischemic retinopathy, retinitis pigmentosa, Chronic and acute glaucoma, macular degeneration, retinal arterial occlusion, Korea, Hungtington's disease, Hungtington's Korea, cerebral edema, proctitis, postherpetic neuralgia, eudinia, heat sensitivity, sarcoidosis, irritable bowel syndrome, Tourette syndrome, Lesch syndrome -Nyhan, Brugado syndrome, Liddle syndrome, Crohns disease, multiple sclerosis and the pain associated with multiple sclerosis (MS), amyotropic lateral sclerosis (ALS), Disseminated sclerosis, diabetic neuropathy, peripheral neuropathy, Charcot Marie dental syndrome, arthritis, rheumatoid arthritis, osteoarthritis, chondrocalcinosis, paroxysmal dystonia, myasthenia syndromes, myotonia, mlotonic dystrophy, muscular dystrophy, cystic fibrosis, cystic fibrosis, mental, bipolar depression, anxiety, schizophrenia, diseases related to toxins in sodium channels, hereditary erythromelalgia, primary erythromelalgia, rectal pain, cancer, epilepsy, partial and general tonic attacks, febrile attacks, absence attacks (petit mal), myoclonic attacks, clonic attacks, Lennox Gastaut syndrome, West

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<img file="MX342298B_D0037.tif" />

(infantile spasms), sick sinus syndrome (Haufe V, Chamberland C, Dumaine R, J. Mol. Cell Cardiol. 42 (3): 469-477 (2007)), multi-resistant attacks, prophylactic (anti-epileptogenic) attacks, hereditary Mediterranean fever syndrome, gout, restless leg syndrome, arrhythmias, fibromyalgia, neuroprotection under ischemic conditions caused by neural attack or trauma, tachyarrhythmias, atrial fibrillation and ventricular fibrillation, and as a general or local anesthetic.

As used herein, the term "pain" refers to all categories of pain, regardless of their nature or origin, and is understood to include, but is not limited to, neuropathic pain, inflammatory pain, nociceptive pain , idiopathic pain, neuralgic pain, orofacial pain, burn pain, chronic bone pain, lower back pain, neck pain, abdominal pain, burning mouth syndrome, somatic pain, visceral pain (including abdominal pain), myofascial pain, dental pain, cancer pain, chemotherapy pain, myofascial pain syndrome, complex regional pain syndrome (CRPS), tempomandibular joint pain, trauma pain, Extreme Paroxysmal Pain Disorder, surgical pain, post-surgical pain, labor pain, labor pain, reflex sympathetic dystrophy, avulsion of the brachial plexus, neurogenic bladder, acute pain, musculoskeletal pain, post-operative pain, chronic pain, persistent pain, peripherally mediated pain, centrally mediated pain, chronic headache, tension headache, cluster headache or vasogenic facial pain (cluster headache),

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<img file="MX342298B_D0038.tif" />

migraine headache, hereditary hemiplegic migraine, conditions associated with headache, sinus headache, phantom limb pain, peripheral nerve damage, post-attack pain, thalamic lesions, radiculopathy, HIV pain, post-herpetic pain, non-cardiac pain Chest, irritable bowel syndrome and pain associated with intestinal disorders and dyspepsia, and combinations thereof.

The present invention also relates to compounds, pharmaceutical compositions, and methods of using compounds and pharmaceutical compositions in the treatment or prevention of diseases or conditions such as benign prosthetic hyperplasia (BPH), hypercholesterolemia, cancer, and pruritus (itching).

Benign Prostatic Hyperplasia (BPH), also known as Benign Prosthetic Hypertrophy, is one of the most common diseases affecting older men. BPH is a progressive condition characterized by a nodular enlargement of the prosthetic tissue that results in obstruction of the urethra. Consequences of BPH may include hypertrophy of the soft bladder muscle, an unbalanced bladder, acute retention of urine, and an increased incidence of urinary tract infections.

BPH has a high impact on public health and is one of the most common reasons for surgical intervention in older men. Attempts have been made to clarify the etiology and pathogenesis, and experimental models have been developed for this purpose. Spontaneous animal models are

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<img file="MX342298B_D0039.tif" />

They imitate the chimpanzee and the dog. The BPH in the hnmhmy.gn ai pprrp share many common characteristics. In both species the development of BHP occurs spontaneously with advanced age and can be prevented with early / prepubertal castration. A medical alternative to surgery is highly desirable for the treatment of BPH and its consequences.

Prosthetic epithelial hyperplasia, both male and dog, is sensitive to androgens: it undergoes involution due to androgen deprivation and resumes epithelial hyperplasia when the androgen is replenished. Cells from the prosthetic gland have been shown to express high levels of voltage-regulated sodium channels. Immunostaining studies clearly demonstrated evidence of voltage-gated sodium channels in prosthetic tissues {Prostate Cancer Prostatic Dis. 2005; 8 (3): 266-73). Inhibition with tetrodotoxin {tetrodotoxin), a selective blocker, of voltage-gated sodium channel function, inhibits migration of prostate-derived cells and breast cancers (Brackenbury, WJ and Djamgoz, MBA, J. Physiol. (Lond) (2006) 573: 343-56; Chloni, AM. et al., Int. J. Biochem. Cell Biol. (2009) 41: 1216-1227).

Hypercholesterolemla, that is, high blood cholesterol value, is an established risk factor in the development of, for example, atherosclerosis, coronary artery disease, hyperlipidemia, seizures, hyperlnsulinemlas, hypertension, obesity, diabetes, cardiovascular diseases ( CVD), mylocardial ischemia, and heart attacks.

<img file="MX342298B_D0040.tif" />

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Thus, lowering total serum cholesterol levels in individuals with high cholesterol levels is known to reduce the risks of these diseases. In particular, the reduction of low-density llpoprotein cholesterol is an essential step in the prevention of CVD. Although there are a variety of therapies against hypercholesterolemia, there is a continuing need and search for alternative therapies in this field of art.

The invention provides compounds that are useful as antihypercholesterolemic agents and their related conditions. The compounds present can act in various ways. Although not intended to be linked to any particular mechanism of action, the compounds may be direct or indirect inhibitors of the enzyme acyl CoA: acyl cholesterol transferase (ACAT) which results in Inhibition of esterification and transport of cholesterol through the intestinal wall. Another possibility is that the compounds of the invention may be direct or indirect inhibitors of cholesterol biosynthesis in the liver. It is possible that some compounds of the Invention act as direct and indirect inhibitors of ACAT and cholesterol biosynthesis.

Pruritus, commonly known as itching, is a common dermatological condition. There are two broad categories of itch based on their etiologies: inflammatory skin itch and neuropathic itch (Blnder et al., Natura Clinical Practice, 4: 329-337, 2008). In the first case, inflammatory mediators activate skin pruriceptors, which

<img file="MX342298B_D0041.tif" />

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INDUSTRIAL are a subset of afferent dermal nerve fibers, primarily C-fibers without myelin. Treatment for this type of itch is either blocking the receptors for inflammatory agents (such as antihistamines) or blocking subsequent electrical activity. Voltage-regulated sodium channels play a central role in the transmission of electrical activity in neurons, and modulation of voltage-regulated sodium channels is a well-known modulation medium for this type of communication. Although the causes of neuropathic pruritus are complex and less understood, there is well-established evidence, from the neuronal sensory C fibers of the dermis, of the existence of central sensitivity and hypersensitivity to influx. Regarding inflammatory pruritus, sodium channels appear to be essential for the propagation of electrical signals from the skin to the central nervous system (CNS). Transmission of the itching impulses produces the uncomfortable feeling that the desire or reflex to scratch generates.

Both inflammatory and neuropathic itching can be blocked by known voltage-regulated sodium channel blockers, with lidocaine being the most common (Villamil et al., American Journal of Medicine 118: 1160-1163, 2005; Inan et al., Euorpean Journal of

Pharmacology 616: 141-146, 2009; Fishman et al., American Journal of Medicine 102: 584-585, 1997; Ross et al., Neuron 65: 886-898, 2010). The lidocaine doses needed to relieve itching are comparable to those effective in treating pain. Both sensory circuits

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INDUSTRIAL share common mediators and related neural pathways (Ikoma et al., Nature Reviews Neuroscience, 7: 535-547, 2006). However, other pain treatments are ineffective against itching and may exacerbate itching rather than alleviate it. For example, oploids in particular, are effective in relieving pain but can cause severe itching. Thus, blocking voltage-gated sodium channels is a particularly promising therapy for both pain and itching.

Compounds of the present invention have been found to have analgesic effects in a number of animal models, in doses ranging from 1mg / Kg to 100mg / Kg. The compounds of the invention may also be useful in treating itching.

Types of itching or skin irritation Include, but are not limited to:

a) psoriatic itching, itching due to hemodialysis, "aquagenic" pruritus (aguagenic), and itching due to skin disorders (eg, contact dermatitis), systemic disorders, neuropathy, psychogenic factors, or a mixture of the previous ones;

b) itching caused by allergic reactions, insect bites, hypersensitivity (eg, dry skin, acne, eczema, psoriasis), inflammatory conditions or injury;

c) itching associated with vulvar vestibulitis;

d) skin irritation or inflammatory effect from the administration of another therapeutic such as, for example, antibiotics,

<img file="MX342298B_D0042.tif" />

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INDUSTRIAL antivirals and antihistamines; and _

e) itching due to activation of the coupled receptors of the G-protein PAR-2.

The (S) -enantiomer of the invention modulates, preferably inhibits, ion flow through a voltage dependent sodium channel. Preferably, the (S) -enantlomer of the invention is a state or frequency dependent modifier of voltage regulated sodium channels, with a low affinity for the limited / closed state and a high affinity for the deactivated state. Although not intended to be linked to any particular mechanism of action, the (S) -enantiomer of the invention will likely interact with the overlapping sites located in the internal cavity of the channel's sodium-conducting pore, similar to that described for other state-dependent sodium channel blockers (Cestéle, S., et al., op. cit.). The (S) -enantiomer of the invention will also likely interact with sites external to the internal cavity and have allosteric effects on sodium ion conduction through the channel pore.

In a preferred embodiment of the invention, the (S) -enantiomer of the invention modulates, preferably inhibits, the activity of Nay1.7. In another preferred embodiment of the invention, the (S) -enantiomer of the invention selectively modulates, preferably inhibits, the activity of Na / I.7 compared to the modulation or inhibition of other voltage regulated sodium channels (/.e .. Na / 1.1 to Na / I.6 and Na_v1.8 to Na / I.9). Because most of the other sodium channels are involved in other processes

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INSTITUTO MBXiCANO • E LA RRURIEDAD

1NDUSTRI / U

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Important physiological factors such as contraction and rhythmity of the heart (Na / 1.5), contraction of skeletal muscle (Na / 1.4), and conduction of electrical activity in the CNS and motor neurons (Na / 1.1, Nay 1.2 and Nay 1.6), it is desirable that the (S) -enantiomer of the invention avoids significant modulation of these other sodium channels.

Any of these consequences may eventually be responsible for the general benefit provided by the (S) -enantiomer of the invention.

Typically a successful therapeutic agent of the invention will meet some or all of the following criteria. Oral availability must be at 20% or more. The efficacy in an animal model is less than about 0.1 pg to about 100 mg / Kg of body weight and the target human dose is between 0.1 pg to about 100 mg / Kg of body weight, although doses outside this range may be acceptable. ("Mg / Kg" means milligrams of the compound per kilogram of body mass of the subject to whom it is being administered). The therapeutic index (or ratio of toxic dose to therapeutic dose) must be greater than 100. The power (expressed by the IC value)₅o) should be less than 10 pM, preferably less than 1 pM, and more preferably less than 50 nM. The IC50 ("Inhibitory Concentration - 50%) is a measure of the amount of the (S) -enantiomer of the invention required to achieve 50% inhibition of ion flow through a sodium channel, over a period of time specific, in an assay of the invention.

Another aspect of the invention relates to the inhibition of

ΙΜΡΙΟ&gt;

INSTITUTO MEXICANO OE LA PROPIEDAD industrial Na activity_v1.1, Nay1.2, Na / I.3, Na / 1.4, Na / 1.5, Nai / I. fi Nad. 7. Na&gt; /1.8. or Nay1.9 in a biological sample or in a mammal, preferably a human, method comprising administering to a mammal or contacting a biological sample with the (S) -enantiomer of the invention or with a composition comprising (S) - enantiomer of the invention. The term "biological sample" as used herein includes, without limitation, cell cultures or extracts thereof; biopsy material obtained from mammals or extracts thereof; and blood, saliva, urine, feces, semen, tears, or other body fluids or extracts thereof.

In addition to the preceding uses of the (S) -enantiomer of the invention, the compound may also be useful in modulating, preferably inhibiting, the activity of voltage-gated sodium channels in a biological sample, for different purposes known to a right-handed in art. Examples of such purposes include, without intending to be limiting, the study of voltage-regulated sodium ion channels in biological and pathological phenomena; and the comparative evaluation of new or other voltage regulated sodium ion channel modulators.

The (S) -enantiomer of the invention can also be used to treat non-human mammals (eg, veterinary treatment methods) for diseases or conditions that are ameliorated or alleviated by modulation, preferably inhibition, of sodium channels regulated by voltage, particularly for the treatment of inflammation and pain. Such

<img file="MX342298B_D0045.tif" />

INSTITUTO MEXICANO DE IA ΓΚΟΡΙΕΓΙΛΓ INDUSTRIAL treatment is understood to be of particular interest to companion mammals, such as dogs and cats.

Pharmaceutical compositions of the invention and its administration

The present invention also relates to pharmaceutical compositions containing the (S) -enantiomer of the invention. In one embodiment, the present invention relates to a composition of the (S) enantiomer of the invention in a pharmaceutically acceptable carrier and in an amount effective to modulate, preferably inhibit, the flow of ions through a voltage regulated sodium channel , to treat diseases, such as pain, when administered to an animal, preferably a mammal, more preferably a human patient.

Administration of the (S) -enantiomer of the invention, in pure form or in a suitable pharmaceutical composition, can be accomplished via any of the accepted forms of administration of agents that serve similar utilities. The pharmaceutical compositions of the invention can be prepared by combining a compound of the invention with a pharmaceutically suitable and acceptable carrier, diluent or excipient, and can be formulated in solid, semi-solid, liquid or gaseous preparations, such as tablets, capsules, powders, granules, ointments , solutions, suppositories, injections, inhalers, microspheres and aerosols. Typical routes for administering such pharmaceutical compositions include,

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MEXICAN INSTITUTE • F THE INDUSTRY PROPERTY!

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Unlimited, oral, topical, transdermal, inhaled, parenteral, sublingual, rectal, vaginal, and intranasal. The term parenteral as used herein includes subcutaneous, intravenous, intramuscular, intra-western (intrastemal) injections, and infusion techniques. The pharmaceutical compositions of the invention are formulated to allow the active ingredients contained therein to be bioavailable upon administration of the composition to the patient. The compositions to be administered to a subject or patient, preferably a mammal, more preferably a human, take the form of one or more dosage units, in which a tablet, for example, may be a single dose, and a container of A compound of the invention in aerosol form can contain a plurality of dosage units. The actual methods of preparing such dosage forms are known, or will become apparent, to those skilled in the art; for example, see The Science and Practice of Pharmacy, 20th Edltion (Philadelphia College of

Pharmacy and Science, 2000). The composition to be administered, in any case, will contain an effectively therapeutic amount of a compound of the invention, or a pharmaceutically acceptable salt thereof, for the treatment of a disease or condition of interest according to the teachings of this invention.

The useful pharmaceutical compositions referred to herein also contain a pharmaceutically acceptable carrier, including any suitable diluent or excipient, including any pharmaceutical agent that does not in itself induce the production of antibodies.

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MEXICAN INSTITUTE BE THE INBIISTRIAL PROPERTY

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harmful to the individual receiving the composition, and which can be administered without undue toxicity. Pharmaceutically acceptable carriers include, without limitation, liquids, such as water, saline, glycerol, ethanol, and the like. A comprehensive discussion of carriers, diluents, and other excipients is presented in REMINGTON'S PHARMACEUTICAL SCIENCES (Mack Pub. Co., NJ, current issue).

A pharmaceutical composition of the invention can be in the form of a solid or liquid. In one aspect, the carrier (s) is / are particulates, such that the compositions are, for example, in the form of tablets or powder. The carrier (s) may be liquid, with the compositions, for example, in the form of an oral syrup, an injectable liquid, or an aerosol, which is useful in, for example, inhaled administration.

When for oral administration, the pharmaceutical composition is preferably in solid or liquid form, where semi-solid, semi-liquid, suspension and gel are forms included within the forms that are considered solid or liquid herein.

As a solid composition for oral administration, the pharmaceutical composition can be formulated into powder, granules, compressed tablets, lozenges, capsules, chewing gum, biscuits or the like. Such a solid composition will typically contain one or more inert diluents or edible carriers. Additionally, one or more of the following may be present: binders such as carboxy methyl methyl cellulose, ethyl cellulose, microcrystalline cellulose, tragacanth gum or gelatin; excipients

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as starch, lactose or dextrins; disintegrating agents such as alginic acid, sodium alginate, Primogel, corn starch and the like; lubricants such as sucrose or saccharin; a flavoring agent such as peppermint, methyl salicylate, or orange flavor; and a coloring agent.

When the pharmaceutical composition is in the form of a capsule, for example a gelatin capsule, it may contain, in addition to materials of the types described above, a liquid carrier such as polyethylene glycol or oil.

The pharmaceutical composition may be in liquid form, for example an elixir, syrup, solution, emulsion, or suspension. The liquid can be for oral administration or for injected application, to name two examples. When the objective is oral administration, the preferred compositions contain, in addition to the (S) -enantiomer of the invention, one or more of a sweetening agent, preservatives, dye / colorant and a flavor improver. In a composition to be injected, one or more of a surfactant, preservative, wetting agent, dispersing agent, suspending agent, chemical buffer (buffer), stabilizer and an isotonic agent can be included.

The liquid pharmaceutical compositions of the invention, whether 20 solutions, suspensions, or the like, can include one or more of the following adjuvants: sterile diluents such as water for injection, saline, preferably physiological solution, Ringer's solution, isotonic sodium chloride, fixed oils such as mono and synthetic diglycerides,

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INSTITUTO MHUCANÜ »&amp; LA norttBAD INDUSTRIAL

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they can serve as a solvent or suspending medium, poltetilpnfliir.nl glycerin, propylene glycol or other solvents; antibacterial agents such as benzyl alcohol or methyl paraben; antioxidants like ascorbic acid or sodium bisulfite; chelating agents such as ethylenediaminetetracetic acid;

chemical buffers such as acetates, citrates, or phosphates, and tonicity-adjusting agents such as sodium chloride or dextrose. The parenteral preparation can be contained in ampoules, disposable syringes or multi-dose ampoules made of glass or plastic. A preferred adjuvant is physiological saline. An injectable pharmaceutical composition is preferably sterile.

A liquid pharmaceutical composition of the invention intended to be administered parenterally or orally must contain an amount of the (S) -enantiomer of the invention such that a suitable dose is obtained. Typically, this amount is at least 0.01% of the (S) -enantiomer of the invention in the composition. When intended for oral administration, this amount may vary between 0.1 and 70% by weight of the composition. Preferred oral pharmaceutical compositions contain between 4% and about 50% of the (S) -enantiomer of the invention. The preferred pharmaceutical compositions and preparations according to the present invention are prepared such that a parenteral dose unit contains between 0.01 to 10% by weight of the (S) -enantiomer of the invention prior to dilution.

The pharmaceutical composition of the invention may be intended for topical administration, in which case the carrier may

<img file="MX342298B_D0050.tif" />

MBXICAN INSTITUTE OF INDUSTRIAL PROPERTY comprise a solution, emulsion, ointment or gel base. The base, for example, can comprise one or more of the following; petrolatum, lanolin, polyethylene glycol, beeswax, mineral oil, diluents such as water and alcohol, emulsifiers and stabilizers. The thickening agents can be present in a pharmaceutical composition for topical administration. If it is for transdermal administration, the composition may include a transdermal patch or an iontophoresis device. Topical formulations may contain a concentration of the (S) -enantiomer of the invention between 0.1 to about 10% by weight by volume.

For topical applications it is preferred to administer an effective amount of a pharmaceutical composition prepared according to the invention to attack an area, eg, skin surfaces, mucous membranes, and the like, that are adjacent to peripheral neurons to be treated. This amount will generally be between about 0.0001 mg to approximately

one g of the (S) -enantiomer of the invention by application, depending on the area to be treated, whether the use is diagnostic, prophylactic or therapeutic, the severity of the symptoms, and the nature of the topical vehicle used. A preferred topical preparation is ointment, in which between 0.001 to about 50 mg of active ingredient is used per cc of base ointment. The pharmaceutical composition can be formulated as transdermal compositions or transdermal delivery devices ("patches"). Such compositions include, for example, a backing, a container of the active ingredient, a control membrane, a coating and contact adhesive.

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MEXICAN INSTITUTE OF PROPERTY

INDUSTRIAL

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These transdermal patches can be used to deliver contiguous pulses to deliver on demand the compounds of the present invention as desired.

The pharmaceutical composition of the invention may be intended for rectal administration in the form, for example, of a suppository, which will melt in the rectum, releasing the drug. The composition for rectal administration may contain an oil base as a suitable non-irritating carrier. Such bases include, but are not limited to, enumeration, lanolin, cocoa butter and polyethylene glycol.

A typical formulation for intramuscular or intra-spinal (intrathecal) administration will consist of a suspension or solution of an active ingredient in an oil, for example arachis oil or sesame oil. A typical formulation for intravenous or intraspinal administration will consist of a sterile isotonic aqueous solution containing, for example, an active ingredient and dextrose or sodium chloride, or a mixture of dextrose and sodium chloride.

The compositions of the invention can be formulated to provide rapid, sustained or delayed release of the active ingredient, i.e. the (S) -enantiomer of the invention, after being administered to the patient using procedures known in the art. Controlled drug delivery systems include osmotic pump systems and liquefaction (dissolutional) systems that contain polymer-coated reservoirs or drug-polymer matrix formulations. Examples of

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INSTITUTO MBXICANO r&gt; f LA PHOPIÍDAD INDUSTRIAL

<img file="MX342298B_D0052.tif" />

controlled release systems are given in US Pat. Nos. 3,845,770 and 4,326,525 and in PJ Kuzma et al., Regional Anesthesia 22 (6): 543-551 (1997), all of which are incorporated herein by reference.

The compositions of the invention can also be administered through intranasal delivery systems in local, systemic and nasocerebral medical therapies. Controlled Particle Dispersion (CPD) ™ technology, traditional nasal spray containers, inhalers or nebulizers, are known to those skilled in the art as effective providers of drug delivery in local and systemic cases, pointing to the olfactory region and the paranasal sinuses.

The invention also relates to a drug-releasing shell or core intravaginal device suitable for administration to a human or animal female. The device may be comprised of the active pharmaceutical ingredient in a polymeric matrix surrounded by a sheath and capable of releasing ei (S) -enantiomer of the invention in a substantially zero order pattern, on a daily basis similar to the devices used to apply testosterone as described in the PCT Patent

Published No. WO 98/50016.

Current eye release methods include topical administration (drops), subconjunctive injections, periocular injections, intravitreal injections, surgical implants, and iontophoresis.

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MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY

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(A small electric current is used to transport ionized drugs into and through the body.) Those skilled in the art would combine the most suitable excipients with the (S) -enantiomer of the invention for a safe and effective infraocular administration.

The most appropriate route of administration will depend on the nature and severity of the condition to be treated. Those skilled in the art are also familiar with determining methods (i.e. oral, intravenous, inhalation, subcutaneous, rectal, etc.), dosage forms, pharmaceutically suitable excipients, and other matters relevant to the release of (S) - enantiomer of the invention to a subject that requires it.

The pharmaceutical composition of the invention can include various materials that modify the physical form of a solid or liquid dosage unit. For example, the composition may include materials that form a coating layer around the active ingredients. The materials that form the coating are typically inert and can be selected from, for example, sugar, shellac and other enteric coating agents. Alternatively, the active Ingredients can be contained in a gelatin capsule.

The pharmaceutical composition of the invention in solid or liquid form may include an agent that amalgamates the (S) -enantiomer of the invention and thereby aids in the release of the compound. Suitable agents that can act in this way include a mono or polyclonal antibody, a protein, or a liposome.

IMPI

MEXICAN INSTITUTE Say THE INDUSTRIAL PROPERTY

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The pharmaceutical composition of the invention-can-eoasisploys. dosage units that can be administered as an aerosol. The term aerosol is used to indicate a variety of systems ranging from those of a colloidal nature to systems consisting of pressurized packaging. Release can be by a liquefied or compressed gas or by a suitable pump system that dispenses the active Ingredients. The aerosols of the (S) -enantlomer of the invention can be released by mono, b, or triphasic systems to release the active ingredient (s). Aerosol release Includes the necessary container, activators, valves, subcontainers, and the like, which together can form a kit / kit (kif). Someone skilled in the art, without much experimentation, can determine the preferred sprays.

The pharmaceutical compositions of the invention can be prepared by methodologies well known in the pharmaceutical art.

For example, a pharmaceutical composition intended to be administered by injection may be prepared by combining the (S) -enantiomer of the invention with sterile distilled water to form a solution. A surfactant can be added to facilitate the formation of a homogeneous solution or suspension. Surfactants are compounds that interact non-covalently with the (S) -enantlomer of the invention to facilitate the homogeneous dissolution or suspension of the compound in the aqueous delivery system.

The (S) -enantlomer of the invention should be administered in a

IJMí Ρ1

INSTITUTO MtXICANC, í&gt; k LA PKOi-'rOArj industrial therapeutically effective amount, which will vary depending on several factors including the activity of the specific compound used; of the metabolic stability and extension of the action of the (S) -enantiomer of the invention; the age, body weight, general health, gender and diet of the patient; the mode and time of administration; the excretion rate; the combination of drugs; the severity of the disorder or particular condition; and of the subject to whom the therapy is applied. In general, a therapeutically effective daily dose of the (S) -enantiomer of the invention is (for a 70 Kg mammal) from about 0.001 mg / Kg (i.e. 0.07 mg) to about 100 mg / Kg (i.e. 7.0 g); preferably a therapeutically effective daily dose is (for a 70 Kg mammal) from about 0.01 mg / Kg (ie 0.70 mg) to about 50 mg / Kg (ie 3.5 g); and more preferably a therapeutically effective daily dose is (for a 70 Kg mammal) from about 1 mg / Kg (ie 70 mg) to about 25 mg / Kg (ie 1.75 g).

The effective dose ranges presented here are not intended to be limiting and represent preferred dose ranges. However, the most preferred dose will be tailored to the individual subject, as understood and determined by someone skilled in the relevant arts, (see (see, eg, Berkowef al., Eds., The Merck Manual, 16^{, h} edition, Merck and Co., Rahway, NJ, 1992; Goodmanetna., Eds., Goodman and Cilman's The Pharmacological Basis of Therapeutics, 10^th edition, Pergamon Press, Inc., Elmsford, NY, (2001); Avery's Drug Treatment: Principles and Practice of Clinical Pharmacology and Therapeutics, 3rd edition, ADIS Press, LTD., Williams and Wilkins, Baltimore,

IMPI

<img file="MX342298B_D0055.tif" />

MD. (1987), Ebadi, Pharmacology, Little, Brown and Co., Boston, (1985); Osolci al., Eds., Remongton's Pharmaceutical Sciences, 18^{, h} edition, Mack Publishing Co., Easton, PA (1990); Katzung, Basic and Clinical Pharmacology, Appleton and Lange, Norwalk, CT (1992)).

<img file="MX342298B_D0056.tif" />

The total dose required for each treatment can be administered in multiple doses or in a single dose over the course of a day, if desired. Generally, treatment is started with smaller doses that are less than the optimal dose of the compound. Thereafter, the dose is increased in small increments until the optimal effect, under the prevailing circumstances, is achieved. The diagnosed compound or pharmaceutical composition may be administered alone or in conjunction with other diagnosed / s and / or drugs directed at pathology, or directed at other symptoms of pathology. Effective amounts of the (S) enantlomer of the invention or of the composition of the invention range from about 0.1 pg to about 100 mg / Kg of body weight, administered at intervals of 4-72 hours, over a period of 2 hours to 1 year , and / or any range or value within those ranges, such as 0.0001-0.001; 0.001-0.01; 0.01-0.1; 0.1-1.0; 1.0-10; 5-10, 10-20, 20-50 and 50-100 mg / Kg, at Intervals 1-4, 4-10, 10-16, 16-24, 24-36, 24-36, 36-48, 48-72 hours, for a period of

1-14, 14-28, or 30-44 days, or 1-24 weeks, or any range or value in between those ranges.

The receptor for administration of the (S) -enantiomer of the invention and / or compositions of the invention can be any animal, 'MPI &lt;· Η-πιτο MEXICANO&gt;' THE INDUSTRIAL PROPERTY

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like a mammal for example. Among mammals the preferred receptors are mammals of the Primates Orders (including humans, apes, and monkeys), Arterlodactyla (including horses, goats, cows, sheep, pigs), Rodenta (including mice, rats, rabbits, and hamsters), and Carnivora ( including cats and dogs). Among birds, the preferred recipients are turkeys, chickens, and other members of the same Order. The most preferred receptors are humans.

Combined Therapy

The (S) -enantiomer of the invention can be usefully combined with one or more therapeutic agents or as a combination thereof, for the treatment of diseases and conditions in mammals, preferably humans, that are ameliorated or alleviated by modulation, preferably inhibition of voltage regulated sodium channels. For example, the (S) -enantiomer of the invention can be administered simultaneously, sequentially, or separately in combination with other therapeutic agents, including, without being taken as limiting;

• opioid pain relievers, such as: morphine, heroin, cocaine, oxymorphine, levorphanol, levalorfan, oxycodone, codeine, dihydrocodeine, propoxyphene, nalmefen, fentanyl, hydrocodone, hydromorphone, meripidine, methadone, nalorphine, naloxone, naloxone, naloxone, naloxone, naloxone, naloxone, naloxone, naloxone, naloxone, naloxone, naloxone, naloxone, naloxone, naloxone, naloxone, naloxone, naloxone, naloxone, naloxone, naloxone nalbuphine and pentazocine;

• non-opioid pain relievers, such as:

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MEXICAN INSTITUTE • E LA ΡλΟΠΕΟΛΓ INDUSTRIAL

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acetaminophen, salicylates (eg aspirin).

• non-steroidal anti-inflammatory drugs (NSAIDs), such as: ibuprofen, naproxen, fenoprofen, ketoprofen, celecoxib, diclofenac, diflusinal, etodolac, fenbufen, fenoprofen, flufenisal, flurbiprofen, ibuprofen, indomethacin, indomethacin, , mefenamic acid, meloxicam, nabumetone, naproxen, nimesulide, nitroflurbiprofen, olsalazine, oxaprozine, phenylbutazone, piroxicam, sulfasalazine, sulindac, tolmetin and zomepirac;

• anticonvulsants, such as: carbamazepine, oxcarbazepine, lamotrigine, valproata, topiramate, gabapentin, and pregabalin;

• antidepressants such as tricyclic antidepressants, such as, for example, amitriptyline, clomipramine, despramine, imipramine and nortriptyline;

• selective COX-2 inhibitors, such as: celecoxib, rofecoxib, parecoxib, valdecoxib, deracoxib, etoricoxib, and lumiracoxib;

• alpha-adrenergics, such as: doxazosin, tamsulosin, clonidine, guanfacine, dexmetatomidine, modafinil, and 4-amino6,7-dimethoxy-2- (5-methane sulfonamide-1,2,3,4-tetrahydroisoquinol-2- yl) -5- (2pyridyl) quinazoline;

• barbiturate sedatives, such as: amobarbital, approbital, butabarbital, butabital, mefobarbital, metarbital, methohexital, pentobarbital, phenobartital, secobarbital, talbutal, teamilal, and thiopental;

ΪΜΡΙ

INSTITUTO MtXICANC O ». IA MOWtlMD INI'USTRIAl

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tachkinin antagonist (tachykinin) (NK, in ingtSSjrp'di liculdi 11 leirte wv— antagonist NK-3, NK-2 or NK-1, for example: (aR, 9 /?) - 7- [3,5b¡s (trifluoromethyl) benzyl)] - 8,9,10,11-tetrahydro-9-methyl-5- (4-methylphenyl) -7/7 [1 , 4] diazocine [2,1-g] [1,7] -naphthyridine-6-13-dione (TAK-637), 5 - [[2 / ?, 3S) -2 - [(1 R) 5 1 - [3,5-bis (trifluoromethylphenyl] ethoxy-3- (4-fluorophenyl) -4-morpholinyl] -methyl] -1,2-dihydro3 / - / - 1,2,4-triazol-3-one (MK-869), aprepitant, lanepitant, dapitant or 3 - [[2metox¡5- (tr¡fluorometox¡) fen¡l] -met¡lam¡no] -2-fen¡lp¡per¡d¡na ( 2S.3S);

• carbon-pitch pain relievers, particularly paracetamol;

• serotonin reuptake inhibitors, such as:

paroxetine, sertraline, norfluoxetine (fluoxetine desmethyl metabolite), metabolite demethylsertraline, '3 fluvoxamine, paroxetine, citalopram, citalopram metabolite desmethylcitalopram, escitalopram, d, l-fenfluramine, femoxetine, ifoxetine, cyanodine, cyanodine, cyanodine fluoxetine;

Norepinephrine reuptake inhibitors (norepinephrine), such as: maprotiline, lofepramine, mirtazepine, oxaprotiline, fezolamine, tomoxetine, mianserin, buproprione, buproprione, hydroxybuproprione, nomifensin, and viloxazine inhibitor (Vivaxain) (Vivalan) norepinephrine such as reboxetine, particularly (SS) -reboxetine, and venlafaxine duloxetine neuroleptic sedatives / anxiolytics;

• dual serotonin-norepinephrine reuptake inhibitors, such as venlafaxine, venlafaxine metabolite O-desmethylvenlafaxine,

<img file="MX342298B_D0060.tif" />

IMPL · 'Νίτπυτο Mexican i • ε industrial property clomipramine, clomipramine metabolite desmethylclomipramine, duloxetine, milnacipran and imipramine;

• acetylcholinesterase inhibitors such as donepezil;

• 5-HT3 antagonists like ondansetron;

• metabotropic glutamate receptor antagonists (mGluR) or allosteric glutamate agonists or enhancers in mGluR's;

• local anesthetics such as mexiletine and lidocaine;

• corticosteroids like dexamethasone;

• antiarrhythmic drugs such as; mexiletine and phenytoin;

• muscarinic antagonists, such as; tolterodine, proplverine, tropsium chloride, darifenacin, solifenacin, temiverin and ipratropium;

• muscarinic agonists or allosteric acetylcholine enhancers in muscarinic receptors;

• cannabinoids or allosteric endorphin enhancers in cannabinoid receptors;

• Vanyloid receptor agonists (eg, resinferatoxin) or antagonists (eg, capsazepine);

• sedatives, such as; glutethimide, meprobamate, methaqualone, and dichloralphenazone;

• anxiolytics such as benzodiasepins;

• antidepressants like mirtasapine;

• topical agents (eg, lidocaine, capsacin, and

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INSTITUTO MMICANO DE LA NOFIEOA · INDUSTRIAL

<img file="MX342298B_D0061.tif" />

resiniferotoxin);

• muscle relaxants such as benzodiasepins, baclofen, carisoprodol, chlorzoxazone, cyclobenzaprine, methocarbamol, and orphrenadine;

• antihistamines or H1 antagonists;

• NMDA receptor antagonists;

• 5-HT receptor agonists / antagonists;

• PDEV inhibitors;

• tramadol®;

• cholinergic pain relievers (nicotinics);

• alpha-2-delta ligands;

• prostaglandin antagonists subtype E2;

• leukotriene B4 antagonists;

• 5-lipoxygenase inhibitors; and • 5-HT3 antagonists.

Diseases and conditions that can be treated and / or prevented using such combinations include, but are not limited to: centrally and peripherally mediated pain, acute, chronic, neuropathic diseases, as well as other diseases with associated pain and other central nervous disorders such as epilepsy, anxiety, depression, and bipolar disease; or cardiovascular disorders such as arrhythmias, atrial fibrillation, and ventricular fibrillation; disorders

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MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY

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neuromuscular such as restless leg syndrome and muscular paralysis and tetanus (Hamann M, Meisler MH, Richter, A Exp. Neurol. 184 (2): 830-838 (2003)); neuroprotection against attacks, neural trauma and multiple sclerosis; and channelopathies such as erltromelalgia and hereditary rectal pain syndrome.

"Combination" as used herein refers to a mixture or exchange of the (S) -enantlomer of the invention with one or more additional therapeutic agents. Unless the context makes something different clear, "combination" may include simultaneous or sequential release of the (S) enantlomer of the invention with one or more therapeutic agents. Unless the context makes something different clear, "combination may Include dosage forms of the (S) -enantlomer of the invention with another therapeutic agent. Unless the context makes something different clear, "combination" may include routes of administration of the (S) -enantiomer of the invention with another therapeutic agent. Unless the context makes something different clear, "combination" may include formulations of the (S) -enantiomer of the Invention with another therapeutic agent. Dosage forms, routes of administration and pharmaceutical compositions include, without limitation, all those described herein.

A combination therapy of the invention includes topical application of the (S) -enantiomer of the invention with an oral agent. The topical application of the (S) -enantiomer of the invention has a very low systemic exposure and has activity that is additive with a number of oral analgesics. Another possible combination therapy includes an oral dose of (S) -

<img file="MX342298B_D0063.tif" />

IMPI

MSXiC / NQ INSTITUTE

OS THE PROPERTY

INDUSTRIAL enantiomer of the invention with an oral agent. Another additional Hp nnmhination therapy of the invention includes a topical application of the (S) -enantiomer of the invention with a topical agent.

The (S) -enantiomer of the invention can be incorporated into 5 compositions to coat an implantable medical device, such as prostheses, artificial valves, vascular grafts, stents and catheters. Accordingly, the present invention, in another aspect, includes a composition for coating an implantable device comprising a compound of the present invention as described above, and a suitable carrier for coating the implantable device. In yet another aspect, the present invention includes an implantable device coated with a composition comprising the (S) -enantiomer of the invention and a suitable carrier for coating the implantable device. Suitable coatings and the general preparation of coated implantable devices are described in US Patents No. 6,099,562; 5,886,026; and 5,304,121.

Parts Kits

The present invention also provides kits containing a pharmaceutical composition of the invention. The kit also includes Instructions for use of the pharmaceutical composition to modulate the activity of ion channels in the treatment of pain, as well as other accessories disclosed herein. Preferably, a commercial package will contain one or more dosage units of the pharmaceutical composition. By

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MEXICAN INSTITUTE • E LA PROriEBAD

INDUSTRIAL

<img file="MX342298B_D0064.tif" />

For example, such a dosage unit may be a sufficient amount for the preparation of an intravenous injection. It will be apparent to those skilled in the art that such light and air sensitive compositions may require special packaging and / or formulation. For example, a package that is opaque to light, and / or sealed in contact with ambient air, and / or formulated with suitable coatings or excipients may be used.

DETAILED DESCRIPTION OF THE INVENTION

The (S) -enantiomer of the invention and the corresponding (R) -enantiomer are prepared by resolution of formula (I), as indicated in the Summary of the Invention, using high pressure chiral liquid chromatography methods or methods of bed motion simulation chromatography, as described later in the Scheme of

Reaction in which "chiral HPLC (chiral HPLC)" refers to high pressure liquid chiral chromatography, and "SMB" refers to bed motion simulation chromatography:

IMPI

REACTION SCHEME

<img file="MX342298B_D0065.tif" />

<img file="MX342298B_D0066.tif" />

<img file="MX342298B_D0067.tif" />

The compound of formula (I) can be prepared by methods set forth in PCT Published Patent No. WO 2006/110917, by methods described herein, or by methods known to those skilled in the art.

Someone with ordinary skill in the art would recognize variations in the Reaction Scheme shown above that would be suitable for the resolution of the individual enantiomers.

Alternatively, the (S) -enantiomer of the formula (I) and the (/?) 15 enantiomer of the formula (\ -R) can be synthesized from materials that are known or easily prepared using processes analogous to those that are known.

Preferably, the (S) -enantiomer of the invention obtained by solving the methods set forth herein is substantially free of the (/?) 20 enantiomer or contains only traces of the (/?) -Enantiomer.

The following Synthetic Examples serve to illustrate the resolution methods set forth by the Reaction Schemes shown above and are not intended to limit the scope of the invention.

SYNTHETIC EXAMPLE 1

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INSTITUTO MtXICAN OR LA PROPIEDAD INDUSTRIAL

<img file="MX342298B_D0068.tif" />

Synthesis of 1 '- {[5- (trifluoromethyl) furan-2-yl] methyl} spiro [furo [2,3 / j [1,3] benzodioxol-7,3'-indole] -2' (T / - /) - one (Compound of formula (I))

<img file="MX342298B_D0069.tif" />

To a suspension of spiro [furo [2,3- / J [1,3] benzodioxol-7,3'10 indole] -2 '(1' / - /) - one (1.0 g, 3.6 mmol), which can be prepared according to the methods set forth in PCT Published Patent No. WO 2006/110917, and cesium carbonate (3.52 g, 11 mmol) in acetone (50 mL) was added 2-bromomethyl-5-trifluoromethylfuran (1.13 g, 3.9 mmol) in one portion, and the reaction mixture was stirred at 55-60 ° C for 16 hours.

After cooling to room temperature, the reaction mixture was filtered and the filtrate was evaporated under reduced pressure. The residue was subjected to a chromatographic column, eluting with ethyl acetate / hexane 1/9 1/1) to provide 1 '- {[5- (trifluoromethyl) furan-2-yl] methyl} spiro [furo [2, 3 /] [1,3] benzodioxol-7,3'-indole] -2 '(1^,/ - /) - ona_I the compound of formula (I), (1.17 g, 76%) as a white solid: mp 139-141 ° C; ¹H NMR (300 MHz, CDCI3) δ 7.32-6.97 (m, 5H), 6.72 (d, J = 3.3 Hz, 1H), 6.66 (s, 1H), 6.07 (s, 1H), 5,905.88 (m, 2H), 5.05, 4.86 (ABq, J_TOb = 16 1 Hz, 2H), 4.91 (d, J = 9.0 Hz, 1H), 4.66 (d, J = 9.0 Hz, 1H); ¹³C NMR (75 MHz, CDCI3) δ 176.9, 155.7, 153.5,

<img file="MX342298B_D0070.tif" />

TI 'TI TO MEXICANO F LA MONEDAD industiual

148.8, 142.2, 141.9, 140.8, 140.2, 139.7, 139.1, 132.1,428 ?, 1? D.7, 12Ú..L. 123.7, 121.1, 120.1, 117.6, 114.5, 114.4, 110.3, 109.7, 103.0, 101.9, 93.8, 80.0, 57.8, 36.9; MS (ES +) m / z 430.2 (M + 1), 452.2 (M + 23); Cal'd for C22H14F3NO5: C, 61.54%; H, 3.29%; N, 3.26%; Found: C, 61.51%; H

3.29%; N, 3.26%.

SYNTHETIC EXAMPLE 2

Resolution of Compound of Formula (I) by means of Chiral HPLC

The compound of formula (I) was resolved to the (S) -enantiomer of the invention and the corresponding (ft) -enantiomer by chiral HPLC under the following conditions:

Column: Chiralcel ® OJ-RH; 20mm ID x 250mm, 5 mic; Lot:

OJRH CJ-EH001 (Daicel Chemica l Industries, Ltd)

Eluent: Acetonitrile / Water (60/40, v / v, Socratic)

Flow rate: 10 mL / min Run time: 60 min

Loading: 100 mg of the compound of formula (I) in 1 mL of acetonitrile

Room temperature

Under the above chiral HPLC conditions, the (/?) Enantiomer of the compound of formula (I), ie (R) -T - {[5 (trifluoromethyl) furan-2-yl] methyl} spiro [furo [2 , 3- / | [1,3] benzodioxol-7,3^,-indole] 68

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MEXICAN INSTITUTE Ok LA PROCEDA · INDUSTRIAL

<img file="MX342298B_D0071.tif" />

2 '(17 - /) - one, was isolated as the first fraction as a white solid; ee (enantiomeric excess)&gt; 99% (OJ-RH analytical grade, 55% acetonitrile in water); mp 103-105 ° C;¹H NMR (300 MHz, DMSO-cfe) δ 7.32-6.99 (m, 5H), 6.71 (d, J = 3.4 Hz, 1H), 6.67 (s, 1H), 6.05 (s, 1H), 5.89 (d, J = 6.2 Hz, 2H), 5.13, 5.02 (ABq, JAB = 16.4 Hz, 2H), 4.82, 4.72 (ABq, Ab = 9.4 Hz, 2H); ¹³C NMR (75 MHz, CDCI3) δ 177.2, 155.9, 152.0, 149.0, 142.4, 142.0, 141.3,

132.0, 129.1, 123.9, 120.6, 119.2, 117.0, 112.6, 109.3, 108.9, 103.0, 101.6,

93.5, 80.3, 58.2, 36.9; MS (ES +) m / z 430.2 (M + 1), [a]_D -17.46 ° (c 0.99,

DMSO). The (S) -enantiomer of the compound of the formula (I), that is, (S) -T10 {[5- (trifluoromethyl) furan-2-yl] methyl} spiro [furo [2,3 - /] [1 , 3] benzodioxol-7,3'-indole] 2 '(rH) -one, was isolated as a second fraction as a white solid; ee (enantiomeric excess)&gt; 99% (OJ-RH analytical grade, 55% acetonitrile in water); mp 100-102 ° C;¹H NMR (300 MHz, DMSO-cfe) δ 7.32-6.99 (m, 5H), 6.71 (d, J = 3.4 Hz, 1H), 6.67 (s, 1H), 6.05 (s, 1H), 5.89 (d, J = 6.3 Hz, 2H),

5.12, 5.02 (ABq, Ab = 16.4 Hz, 2H), 4.82, 4.72 (ABq, Ab = 9.4 Hz, 2H); ¹³C

NMR (75 MHz, CDCI₃) δ 177.2, 155.9, 152.0, 149.0, 142.4, 142.0, 141.3,

132.0, 129.1, 123.9, 120.6, 119.2, 117.0, 112.6, 109.3, 108.9, 103.0, 101.6,

93.5, 80.3, 58.2, 36.9; MS (ES +) m / z 430.2 (M + 1), [_to]_D -14.04 ° (c 0.99,

DMSO).

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MEXICAN INSTITUTE

OF THE PROPERTY

INDUSTRIAL

SYNTHETIC EXAMPLE 3

<img file="MX342298B_D0072.tif" />

Resolution of Compound of Formula (I) by SMB Chromatography

The compound of formula (I) was resolved to the (S) -enantiomer of the invention and the corresponding (ft) -enantiomer by SMB chromatography under the following conditions:

Extract: 147.05 mL / min

Refined: 76.13 mL / min (Raffinate)

Eluent: 183.18 mL / min

Loading: 40 mL / min

Recycle: 407.88 mL / min

Run time: 0.57 min

Temperature: 25 ° C

Pressure: 46 bar

The loading solution (25 g of the compound of formula (I) in 1.0

L mobile phase (25: 75: 0.1 (v: v: v), acetonitrile / methanol / trifluoroacetic acid mixture)), was continuously injected into the SMB system (Novasep Licosep Lab Unit), equipped with eight identical columns in configuration 2 -2-2-2 containing 110 g (per column, 9.6 cm, 4.8 cm Internal Diameter) of

ChiralPAK-AD as a stationary phase. The first eluted enantiomer, (the (/?) -Enantiomer of the compound of formula (I)) was contained in the raffinate stream and the second eluted enantiomer (the (S) -enantiomer of the compound of formula (I)) was contained in the extraction stream. THE MEXICAN INSTITUTE 'V THE INDUSTRIAL PROPERTY

<img file="MX342298B_D0073.tif" />

Characterization data of the (S) -enantiomer and (ffl-enantiomer obtained from SMB resolution were identical to those obtained previously using chiral HPLC.

The compound of formula (I) was resolved into its 5 constituent enantiomers in a preparative LCMS Waters self-purification system. The first eluted enantiomer from the chiral column was brominated (at a site far enough from the stereogenic center) to achieve the 5'-bromine derivative, which was subsequently crystallized to generate a single crystal suitable for X-ray crystallography. The crystal structure of this brominated derivative of the eluted first enantiomer was obtained and its absolute configuration turned out to be the same as the (R) -enantiomer of the invention. Accordingly, the second eluted enantiomer from the chiral column is the (S) -enantiomer of the invention. Furthermore, the material obtained from the SMB resolution extraction stream had a specific optical rotation of the same sign (positive, that is, clockwise) as that of the material obtained from the aforementioned LC resolution.

Biological tests

Various techniques are known in the art to test the activity of the compound of the invention or determine its solubility in known pharmaceutically acceptable excipients. In order that the invention described herein may be more easily understood, the following biological tests are set forth. It should be understood that these examples are for illustrative purposes only and are not

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MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY

<img file="MX342298B_D0074.tif" />

they should in no way be taken as limiting this Invention.

BIOLOGICAL TEST 1

Guanidine inflow assay (in vitro assay)

This example describes an in vitro assay to test and profile test agents against human or rat voltage regulated sodium channels that are stably expressed in cells of an endogenously or heterogeneously expressed origin. The assay is also useful in determining the IC50 of a modulating, preferably blocking, compound of a voltage regulated sodium channel. The assay is based on guanidine influx described by Reddy, NL, et al., J. Med. Chem. (1998), 41 (17): 3298-302.

The guanidine influx assay is a radiotracer influx assay used to determine the ionic flow activity of voltage regulated sodium channels in a high throughput microplate based format. The assay uses guanidine hydrochloride-¹⁴C in combination with several known voltage regulated sodium channel modulators that produce a continuous inflow, to test the potency of test agents. Potency is determined by an IC50 calculation. Selectivity is determined by comparing the potency of the compound for the voltage-regulated sodium channel of interest with its potency against other voltage-regulated sodium channels (also called

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INSTITUTO MBdCANO ÍJt THE PROPERTY

INDUSTRIAL selectivity ').

<img file="MX342298B_D0075.tif" />

Each of the test agents is tested against cells expressing the voltage regulated sodium channels of interest. Voltage regulated sodium channels are characterized as sensitive or insensitive to TTX. This property is useful when evaluating the activities of a voltage regulated sodium channel of interest when it resides in a population mixed with other voltage regulated sodium channels. Table 1 below summarizes the cell lines that are useful for filtering for some voltage-gated sodium channel activity in the presence or absence of TTX.

TABLE 1

<td>Cellphone line</td><td>MRNA expression</td><td>Functional Characterization</td>

<td>CHO-K1 (Chinese Hamster Ovary; recommended host cell line) ATTC accession number CCL-61</td><td>• Expression of Na_v1.4 shown by RT-PCR • No other expression of Na ^ was detected</td><td>• Increased 18 to 20 times the inflow of [¹⁴C] guanidlna was completely blocked using TTX. (Na_v1.4 is a TTX sensitive channel).</td>

<td>L6 (rat myoblastic cell) ATTC Number CRL-1458</td><td>• Expression of Na / I.4y 1.5</td><td>• 10 to 15 times increase the inflow of [¹⁴C] guanidine was only partially blocked by TTX at 100 nM (the Na_v1.5 is TTX resistant)</td>

<td>SH-SY5Y (human neuroblastoma) ATTC Number CRL-2266</td><td>• Expression of Na / 1.9 and Na / 1.7 published (Blum etal.)</td><td>• Increased from 10 to 16 times the inflow of [¹⁴C] guanidine was partially blocked by TTX (Na_vone 9 is TTX resistant)</td>

<td>SK-N-BE2C (an ATCC human neuroblastoma cell line Number CRL-2268)</td><td>• Expression of Na / I.8</td><td>• Stimulation of BE2C cells with pyrethroids results in a 6-fold increase in inflow of [¹⁴C] guanidine above the reference (background). • TTX partially blocked inflow (Na_v1.8 is TTX resistant)</td>

INSTITUTO MEXICANO OE LA PROPIEDAD industrial

<img file="MX342298B_D0076.tif" />

<td rowspan="2">PC12 (rat pheochromocytoma) ATTC Number CRL-1721</td><td rowspan="2">• Expression of Nav1.2 and NaV1.7</td><td>• The increase dq_m§ 12 times the inflow</td>

<td>[14C] guanidine was completely blocked using TTX. (Nav1.2 and NaV17 are channels sensitive to TTX)</td>

<td>HEK293 (human embryonic kidney) ATTC Number CRL1573</td><td>• Expression of hNaV1.7</td><td>• Nav1.7 is a TTX sensitive channel. The TTX IC50 in the Guadinio functional assay is 8 nM.</td>

It is also possible to use immortalized cell lines that heterogeneously express voltage regulated sodium channels. Stable cloning, transfection, and propagation of such cell lines are known to those skilled in the art (see, eg, Klugbauer, N, et al., EMBO J. (1995), 14 (6): 1084- 90; and Lossin, C., et al., Neuron (2002), 34, pp. 877-884).

Cells expressing the voltage-regulated sodium channels of interest are cultured according to the supplier or in the case of a recombinant cell, in the presence of selective culture media such as G418 (Gibco / lnvitrogen). Cells are disassociated from the culture dishes with an enzyme solution (1X) Trypsin / EDTA (Gibco / lnvitrogen) and analyzed to establish their density and viability using a hemocytometer (Neubauer). Dissociated cells are washed and resuspended in their culture medium, then placed in Poly-D-Lysine (Perkin Elmer) coated Scinti dishes (Scintiplates) (approximately 100,000 cells / well) and incubated at 37 ° C / 5% CO2 for 20-24 hours. After extensive washing with low sodium saline with a HEPES chemical buffer (HEPES74

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MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY buffered (LNHBSS) (150 mM choline chloride, 20 nM HEPES (Sigma), 1 mM

<img file="MX342298B_D0077.tif" />

Calcium Chloride, 5mM Potassium Chloride, 1mM Magnesium Chloride, 10mM Glucose) Test agents are diluted with LNHBSS and then added to each well at the desired concentration. (Varied concentrations of the test agent can be used.) The radiolabelled activation / mixture contains an alkaloid such as veratridine or Aconitine (Sigma) or a pyrethroid such as delmatrine, the scorpion venom Leiurus quinquestríatus hebraeus (Sigma), and¹⁴Guanidine C-hydrochloride (ARC), to measure flow through voltage regulated sodium channels.

After loading the cells with the test agent and the radiolabelled activation / mixture, the Scinti dishes coated with Poly-D-Lysine are extensively washed with LNHBSS supplemented with Guanidine (Sigma). Scinti dishes coated with Poly-D-Lysine are dried and then counted using a Wallac MicroBeta TriLux (Perkin-Elmer Life Sciences). The ability of the test agent to block the activity of voltage regulated sodium channels is determined by comparing the amount of¹⁴Cguanidine present within cells expressing the different voltage-regulated sodium channels. Based on these data, a series of calculations, discussed elsewhere in this specification, can be performed to determine if a test agent is selective for a particular voltage regulated sodium channel.

The IC50 value of a test agent for a specific voltage regulated sodium channel can be determined using the above

IMPI

INSTITUI^-MEXICANO DE LA PI'.OPISDAL INDUSTRIAL

<img file="MX342298B_D0078.tif" />

general method. IC50 can be determined using a 3, 8, 10, 12 or 16 point curve, in duplicate or triplicate, with an initial concentration of 1, 5 or 10 μΜ diluted in series with a final concentration that reaches the sub-nanomolar ranges, nanomolar and low micromolar. Typically, the midpoint concentration of the test agent is set at 1 µΜ, and sequential concentrations of higher or lower half-dilutions are applied (eg. 0.5 μΜ; 5 µΜ and 0.25 µΜ; 10 µΜ and 0.125 µΜ; 20 μΜ etc.). The IC50 curve is calculated using the 4 Parameter Logistic Model or the Sigmoidal Dose-Response Model formula (fit = (A + ((BA) / (1 + ((C / x)^TOD)))).

The increase in selectivity, selectivity factor or selectivity multiple, is calculated by dividing the IC value₅or the test voltage regulated sodium channel between the reference voltage regulated sodium channel, for example, Na / I .5.

According to the above, the compound of formula (I), the (S) enantiomer of the compound of formula (I), that is, the (S) -enantiomer of the invention, and the (R) -enantlomer of the Compound of formula (I), when tested in this test, demonstrated voltage-gated sodium channel blocking activity against hNa / l.7 as shown in Table

2:

TABLE 2

<img file="MX342298B_D0079.tif" />

IMPI '^nSTOTÍÍof,^Ceoad • INDUSTRIAL

<td>Compound</td><td>Chemical name '</td><td>ICso (pM)</td>

<td>(i)</td><td>1 '- {[5- (trifluoromethyl) furan-2- il] methyl) spiro [furo [2,3 - /] [1,3] benzodioxol- 7,3'-indole] -2 '(1'H) -one</td><td> 0.007</td>

<td>(lR)</td><td>(/?) - 1 '- {[5- (trifluoromethyl) furan-2- il] methyl} spiro [furo [2,3- / j [1,3] benzodioxol- 7.3^,-indole] -2^,(1'H) -one</td><td> 4.200</td>

<td>(lS)</td><td>(S) -1 '- {[5- (trifluoromethyl) furan-2- il] methyl} spiro [furo [2,3 - /] [1,3] benzodioxol- 7,3'-indole] -2 '(1^,H) -one</td><td> 0.003</td>

The concentration-response relationship for the (S) -enantiomer of the invention and the (F?) -Enantiomer are shown in FIG. 1. Solid curves indicate the best fit of least squares to a 1: 1 binder isotherm; The IC50 values ​​that describe these curves are given in Table 2. In this model, the (S) -enantiomer of the invention demonstrated significantly greater potency (i.e., a multiple&gt; 100) against hNayl.7 when compared to the inhibitory potency of the corresponding (ft) -enantiomer.

These results favor the use of the (S) -enantiomer of the invention over the (/?) -Enantiomer or the compound of formula (I) (the racemic mixture) for the applications described here because a greater pharmacological activity can be obtained by lower dosage levels with the possibility of fewer side effects. Furthermore, the (F?) - enantiomer is a very important tool for safety studies because it allows to distinguish between effects produced by mechanisms (those mediated by the

IMPI

INSTITUTO MEXICANO DF LA PROPIEDAD INDUSTRIAL

<img file="MX342298B_D0080.tif" />

blocking sodium channels) and off-target activities that can be eliminated in analogues without compromising efficacy. If an adverse effect is the product of a mechanism, then the (S) -enantiomer will be much more potent than the (R) -enantiomer, since the secondary sites of action probably do not have identical stereoselectivities and the two enantiomers probably have similar effects, including potency, on secondary sites of action.

BIOLOGICAL EXAMPLE 2 Electrophysiological test (in vitro test}

HEK293 cells expressing hNa / l.7 were cultured in DMEM media (Gibco) with 0.5 mg / mL G418, +/- 1% PSG, and 10% heat inactivated fetal serum at 37 ° C and 5% CO2 . For the electrophysiological recordings, the cells were placed in 10 mm dishes.

Whole cell records were examined with established voltage clamp methods (Bean et al., Op. Cit.) Using an Axopatch 200B amplifier and Clampex software (Axon Instruments, Union City, CA). All experiments were carried out at room temperature. The electrodes were fire polished for 2-4 Mohms resistors. Voltage errors and capacitance artifacts were minimized by compensation with series resistors and compensation capacitance, respectively. Data was acquired at 40 kHz and filtered

<img file="MX342298B_D0081.tif" />

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OF THE PROPERTY

HDUSTRIAL at 5 kHz. The external solution (bath) consisted of NaCl (140mM), KCI (5mM), CaCb (2mM), MgCI₂ (1 mM), HEPES (10 mM) at pH 7.4. The internal solution (pipette) consisted of (in mM): NaCl (5), CaCI₂ (0.1), MgCI₂ (2), CsCI (10), CsF (120), HEPES (10), EGTA (10), at pH 7.2.

To estimate the steady-state affinity of compounds for the resting and inactivated state of the channel (K_r and K, respectively), test pulses of 12.5 ms at depolarizer voltages from -60 to +90 mt / from a sustained potential of -120 ml / were used to construct the current-voltage relationships (lV curves). A voltage near the peak of the I-ν 'curve (-30 mVaO mi /) was used as the test pulse throughout the remainder of the experiment. The continuous state inactivation curves (availability) were then constructed by measuring the activated current during a test pulse of 8.75 ms after conditioning pulses of 1 second to potentials in the range of -120 to -10 mi /.

Continuous voltage dependence of the bond of a compound to a voltage regulated sodium channel was determined by measuring the blocking of the ion current under two sustained potentials. The link to the channels in idle state was determined using a sustained potential of -120 mV, so that maximum availability was obtained. The link to 20 channels in the unactivated state was evaluated under a sustained potential such that only about 10% of the channels were available to open. The membrane potential was maintained at this voltage for at least 10 seconds so that the drug bond could be balanced.

<img file="MX342298B_D0082.tif" />

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INDUSTRIAL

The apparent dissociation constant at each vo | t? Jfi r * =&gt; raicuiA—. using the equation;

% inhibition = [Drug] X 100 ([Drug] + K_d) where Kd is the dissociation constant (K_r or Ki), and [Drug] is the concentration of the test compound.

Accordingly, when the compound of formula (I), the (S) -enantiomer of the compound of formula (I), i.e., the (S) -enantiomer of the invention, were tested in this model, and the (R) -enantiomer of the compound of formula (I), showed affinities for the resting / closed state and the inactivated state of hNa / l.7, as shown in Table 3 below:

TABLE 3

<td>Compound</td><td>Chemical name</td><td>K, (μΜ)</td><td>K_r (μΜ)</td>

<td>OR)</td><td>1 '- {[5- (trifluoromethyl) furan-2- il] methyl} spiro [furo [2,3-f | [1,3] benzodioxol- 7,3'-indole] -2 '(1'H) -one</td><td> 0.142</td><td>&gt; 10uM</td>

<td>(lR)</td><td>(R) -1 '- {[5- (trifluoromethyl) furan-2- i I] metí} sp i ro [furo [2,3 - /] [1,3] benzod ioxol- y.S'-indoleJ-ZO'HJ-one</td><td> 0.869</td><td>&gt; 10uM</td>

<td>(lS)</td><td>($) - 1 '- {[5- (trifluoromethyl) furan-2- L] met¡l} spiro [furo [2,3 - /] [1,3] benzodioxol- 7,3'-indole] -2 '(1'R) -one</td><td> 0.161</td><td>&gt; 10uM</td>

As these results show, the (S) -enantiomer of the invention is a hNa / l.7 modifier depending on the state and the voltage,

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MEXICAN INSTITUTE OF THE PtOPIK'AU INDUSTRIAL

<img file="MX342298B_D0083.tif" />

with a low affinity for the resting / rest state and a high affinity for the inactive state. The results showed that the (S) -enantlomer was about 5 times more potent in binding to the Inactive state of hNa / 1.7 than the (/?) -Enantiomer. Furthermore, the results showed that the (S) 5 enantlomer is the main responsible for the potency of the racemic mixture, that is, the compound of formula (I).

BIOLOGICAL EXAMPLE 3

In vivo tests

Acute Pain (Formalin Test)

The formalin test is used as an animal model for acute pain. In the formalin test, animals are briefly habituated to a plexiglass test chamber for 20 minutes the day before the experiment. On the day of testing, the animals are randomly injected with the test items. Within 30 minutes of drug administration, 50 pL of 10% formalin is injected subcutaneously into the plantar surface of the hind paw of rats. The acquisition of video data starts immediately after the administration of formalin, with a duration of 90 minutes.

Images are captured using Actimetrix software

Limelight that saves files under the * .ll¡¡ extension, and then converts them to MPEG-4 encoding. The videos are then analyzed using the

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INSTITUTO MIXICANO DF LA PROPIEDAD INDUSTRIAL

<img file="MX342298B_D0084.tif" />

behavioral analysis software The Observer 5.1, (Version 5.0, Noldus Information Technology, Wageningen, The Netherlands). The analysis of the video is carried out observing the behavior of the animal and marking each one according to its type, and defining the extension of the behavior (Dubuisson and Dennis, 1977). The behaviors listed include: (1) normal behavior; (2) do not put weight on the leg; (3) raise the leg; (4) lick / bite and scratch his paw. Lifting, favoring, licking, biting, or scratching the injected paw excessively indicates a response to pain. An analgesic response or protection against the compounds is evidenced if both legs rest on the floor, without any favor, excessive licking, biting or scratching the injected leg.

The analysis of the formalin test data is performed based on two factors: (1) Maximum Percent Potential Inhibitory Effect (% MPIE, in English) and (2) pain rating. The% MPIE is calculated in several steps, where the first is to add the extension of the non-normal behaviors (behaviors 1,2, 3) of each animal. A unique value for the vehicle group is obtained by averaging all the scores within the treatment vehicle group. The following calculation results in an MPIE value for each animal:

MPIE (%) = 100 - [(sum of treatment / vehicle average value) X 100%]

The pain rating is calculated from the weighted scale described above. The duration of the behavior is multiplied by the

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INSTITUTO MEXICANO • E LA WOFIRBAP INBOSTUIAL

<img file="MX342298B_D0085.tif" />

weight (this ranks the severity of the response), and then it is divided by the total extension of the observation to determine the pain level of each animal. The calculation is represented by the following formula:

Pain score = [0 (To) + 1 (T1) + 2 (T2) + 3 (T3)] / (To +

T1 + T2 + T3)

CFA-Induced Chronic Inflammatory Pain

This test evaluates tactile allodynia with calibrated von Frey filaments. After a full week of air conditioning in the

Nursery facilities, 150 μ 150 of the Complete Freund's Adjuvant (CFA) emulsion (CFA suspended in an emulsion (1: 1) oil / saline saline at a concentration of 0.5 mg / mL) were injected subcutaneously into the plantar surface of the left hind legs of rats under mild anesthesia with isoflurane. Animals were allowed to recover from anesthesia and the thermal and mechanical basis nociceptive thresholds of all animals were assessed one week after CFA administration. All animals were habituated to the experimental team for 20 minutes the day before the start of the experiment. Test and control items were administered to animals, and nociceptive thresholds were measured at specific points in time after drug administration to determine analgesic responses to each of the six available treatments. The points in time were determined to show the greatest analgesic effect for each compound.

IMPIttgg

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INDUSTRIAL ------- The (S) -enantiomer of the invention and the corresponding (/?) Enantiomer were compared using oral and topical dosages. FIG. 2 shows a comparison of the efficacy of the (S) -enantiomer of the invention and the (/?) -Enantiomer with oral dosage. Each enantiomer was dosed at 10,

30, 100 or 200 mg / Kg. The plasma concentration obtained with each dose was also determined, and the reversal response to pain (as the percentage increase from the baseline threshold) was plotted as a function of the plasma concentration.

The (S) -enantiomer had a greater maximum effect when dosed at 200 mg / Kg. The (/?) - enantiomer achieved a much higher plasma concentration at an equivalent level dose. This was an unexpected and unusual finding. As a consequence, the use of the racemic mixture, that is, the compound of formula (I), would result in a 10-fold excess of the inactive enantiomer, that is, the (R) -enantiomer. Accordingly, the use of the (S) 15 enantiomer of the invention would greatly increase the possibility of achieving efficacy with a minimal probability of encountering unanticipated activities other than stereoselectives.

The (S) -enantiomer of the invention was also administered topically to animals in varying doses (1%, 2%, 4% and 8% (w / v)) and the nociceptive thresholds were measured at defined points in time after of the administration of the drug, to determine the analgesic responses to each of the available treatments. The points in time were previously determined so as to show the analgesic effect

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<img file="MX342298B_D0086.tif" />

higher for each test compound. _ ______

The response thresholds of the animals to tactile stimuli were measured using a Model 2290 Electrovonfrey anesthesiometer (IITC Life

Science, Woodland Hills, CA) after the Hargreaves test. Animals were placed in an elevated Plexiglass enclosure placed on a metal mesh surface. After 15 minutes of accommodation, a previously calibrated von Frey hair was applied perpendicularly to the sole of the hind legs on the same side of the animals, with sufficient force, measured in grams, to produce a firm response of the leg. The response indicated withdrawal of the painful stimulus and that was the end point of efficacy. The tests continued until determining which hair had the least force to induce a leg reaction or until the cutting force was 20 g. This cutting force is used because it represents approximately 10% of the animal's weight and serves to prevent the complete lifting of the limb due to the use of stiffer hairs, which would change the nature of the stimulus. Data were expressed as the percentage increase from the base threshold, in grams.

The (S) -enantiomer of the invention, when tested with this model, demonstrated an analgesic effect as shown in Table 4:

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<img file="MX342298B_D0087.tif" />

TABLE 4

<td></td><td colspan="4">Percentage increase from baseline (CFB)</td>

<td>Compound</td><td>1% topical</td><td>2% topical</td><td>4% topical</td><td>8% topical</td>

<td>(lS)</td><td> 0.62</td><td> 16.71</td><td> 28.79</td><td> 45.06</td>

The (S) -enantlomer of the Invention at 2%, 4% and 8% (w / v) showed increases in mechanical leg removal thresholds in the von Frey tests, values ​​that are expressed as a percentage increase from of the baseline (IFB) to indicate an analgesic effect. The analgesic effect of (S) -enantlomer increased with increasing doses until reaching the highest dose used of 8% w / v, for which the IFB with the highest percentage was shown at + 45.1%. However, the 1% (w / w) dosing group did not demonstrate an appreciable increase in terms of mechanical leg removal threshold. The results indicate that the (S) -enantiomer has analgesic effects in the CFA-induced Inflammatory pain model in the range of 2% to 8% (w / v).

Post-operative Non -ception Models

In this model, hyperalgesia caused by an intraplanar incision in the paw is measured by applying an incremental tactile stimulus to the paw until the animal withdraws the paw from the applied stimulus. While the animals were anesthetized with 3.5% isoflurane, which is applied via a nasal cone, a longitudinal incision was made with a No. 10 scalpel in the

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INDUSTRIAL plantar of the left hind leg through the skin and fascia, starting 0.5 cm from the proximal heel edge and extending in the direction of the toes. After the incision, the skin was joined using 2 sterile 3-0 silk sutures. The wound site was covered with Polysporin and Betadine. Animals were returned to their cage for recovery until the following day.

Animal withdrawal thresholds to tactile stimuli for both operated (ipsilateral) and non-operated (contralateral) paws can be measured using the Model 2290 anesthesiometer

Electrovonfrey (IITG Life Science, Woodland Hills, CA). Animals were placed in an elevated Plexiglass enclosure placed on a metal mesh surface. After at least 10 minutes of acclimatization, previously calibrated von Frey hairs were applied perpendicularly to the plantar surface of both legs of the animals, with enough force to produce a slight movement of the hair against the plantar surface, in ascending order starting with 10g hair. Testing continued until the hair with the least force was found to produce a rapid movement of the leg or until the cutting force was approximately 20 g. This cutting force is used because it represents approximately 10% of the animal's body weight and serves to prevent the complete lifting of the limb due to the use of stiffer hairs, which would change the nature of the stimulus.

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MEXICAN INSTITUTE

OF THE PROPERTY *·'

INDUSTRIAL

Neuropathic pain model; chronic constriction injury For this model, a 3 cm incision was made through the skin and fascia in the middle of the thigh of the animal's left rear leg, using a No. 10 scalpel. The left sciatic nerve was exposed by means of a Direct dissection of the biceps femoris, with care to minimize bleeding. Four loose ligatures were tied along the sciatic nerve using 4-0 sterile, nondegradable, silk sutures at intervals of 1 to 2mm from each other. The tension of the loose ligatures was tight enough to induce a slight constriction of the sciatic nerve, visible under the dissecting microscope with 4x magnification. In the falsely operated animal the left sciatic nerve was exposed without further manipulation. Antibacterial ointment was applied directly to the wound and the muscle was closed using sterile sutures. Betadine was applied to the muscle and its surroundings, followed by closure of the skin with surgical staples.

Animal withdrawal thresholds to tactile stimuli were measured using the Model 2290 Electrovonfrey anesthesiometer (IITC Life Science, Woodland Hills, CA). Animals were placed in an elevated Plexiglass enclosure placed on a muddy mesh surface. After at least 10 minutes of acclimatization, previously calibrated von Frey hairs were applied perpendicularly to the plantar surface of both legs of the animals, with enough force to produce a slight movement of the hair against the plantar surface, in ascending order starting with the

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INSTITUTO MBXICANO DE LA ΡΙΟΡΙΒΠΑΒ INDUSTRIAL 0.1 g hair. Testing continues until the hair with the least force that produces a rapid movement of the leg is determined or until the cutting force of approximately 20 g is reached. This cutting force is used because it represents approximately 10% of the animal's body weight and serves to prevent the complete lifting of the limb due to the use of stiffer hairs, which would change the nature of the stimulus.

The nociceptive thermal thresholds of the animals were evaluated using the Hargreaves test. After the measurement of the tactile thresholds, the animals were placed in a plexiglass enclosure placed on a raised glass platform with heating units. The glass platform was thermostatically controlled at an approximate temperature of 24-26 ° C for all test runs. Animals were allowed to settle for 10 minutes after entering the compound until all exploratory activity ceased. The Model 226 Plantar / Tail Stimulator Analgesia Meter (IITC, Woodland Hills, CA) analgesia meter was used to apply a radiant heat beam from under the glass platform to the plantar surface of the hind legs. During all test runs, the resting intensity and the active intensity of the heat source were set to 1 and 55 respectively, and a cutoff time of 20 seconds was used to avoid tissue damage.

The (S) -enantiomer was compared with the corresponding (R) enantiomer and the racemic mixture (the compound of formula (I)) in this CCI model using topical application of the drug, as described for the

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IMPI tNSTIT '&lt;TO MEXICANO&gt; r THE PROPERTY »· Οίΐςτ * ΐΛΐ CFA model (see FIG. 3). Each test compound was administered as a 2% (w / v) containing ointment. Consistent with the different activities of these two voltage-regulated sodium channel inhibitor enantiomers, only the (S) -enantiomer of the invention reversed pain responses while the (/?) - enantiomer had no significant increase over the line base. Both the (S) -enantiomer and the racemic mixture show a similar percentage increase with respect to the baseline, which tends to suggest that the (S) -enantiomer is responsible for the analgesic effect.

BIOLOGICAL EXAMPLE 4

Aconltine-Induced Arrhythmia Assay

The antiarrhythmic activity of the compounds of the invention is demonstrated by the following test. Arrhythmia is caused by intravenous administration of aconitine (2.0 pg / Kg) dissolved in physiological saline. Test compounds of the invention are administered intravenously 5 minutes after the administration of aconitine. The evaluation of antiarrhythmic activity is performed by measuring the time from the administration of aconitine to the occurrence of extrasystoles (ES) and the time between the administration of aconitine until the occurrence of ventricular tachycardia (VT).

In rats under isoflurane anesthesia (1/4 to 1/3 of 2%),

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MRXICAN J INSTITUTE

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INDUSTRIAL performs a tracheotomy by first making an incision in the neck area, then isolating the trachea to make a 2mm incision where a 2 cm tracheal tube is inserted into the trachea so that the tube opening is positioned just above mouth. The tube is secured with sutures and attached to a ventilator for the duration of the experiment.

Then 2.5 cm incisions are made in the femoral areas and using a blunt dissection probe, the femoral vessels are isolated. Cannulas are inserted into both femoral veins, one to maintain anesthesia with pentobarbltal (0.02-0, 05 mL) and the other for infusion and injection of the drug and the vehicle. A cannula is inserted into the femoral artery with the transmitter's blood pressure gel catheter.

The ECG connections are attached to the chest muscles in connection position II (upper right / above heart, white tip; bottom left / below heart, red tip). The tips are secured with sutures.

All surgical areas are covered with wet gauze with 0.9% saline. This saline solution (1-1.5 mL at 0.9%) is supplemented to moisten the post-surgery areas. The animal's ECG and its ventilation are left to equilibrate for at least 30 minutes.

The arrhythmia is induced with an aconidine infusion at 2 pg / Kg / min for 5 minutes. During this time the ECG is recorded and continuously observed.

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BIOLOGICAL EXAMPLE 5

Ischemia-Induced Arrhythmia Assay

Rodent models for ventricular arrhythmias, 5 both in acute cardiac version and in prevention paradigms, have been used to test potential therapies for atrial and ventricular arrhythmias in humans. Cardiac ischemia leading to myocardial infarctions is a common cause of morbidity and mortality. The ability of a compound to prevent ischemia-induced ventricular tachycardia and fibrillation is an accepted model for determining the efficacy of a compound in a clinical situation of ventricular tachycardia and ventricular fibrillation.

Anesthesia is first induced with pentobarbital (ip) and maintained by a single intravenous infusion. Male SD rats have a cannula inserted into the trachea to provide artificial ventilation with room air at a volume per stroke of 10 mL / Kg, 60 strokes / minute. The right femoral artery and vein are cannulated using a PE50 tube to record arterial blood pressure (MAP) and intravenous administration of compounds, respectively.

The chest is opened between ribs 4 and 5 to create an opening 20 such that the heart can be seen. Each rat is placed on a slotted platform and metal retainers are attached to the rib cage to open the chest cavity. A suturing needle is used to penetrate the ventricle just below the raised atrium and is drawn diagonally down from

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MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY so that an occlusion zone (OZ) of&gt; 30% and&gt; 50% can be obtained. The exit point is ~ 0.5 cm below where the aorta connects to the left ventricle. The suture is tied so that a loose loop forms around a branch of the artery.

The animal's chest is then closed allowing the end of the occluder to be accessible from outside the chest.

Electrodes are placed in connection position II (right atrium to apex) for ECG measurements, as follows: one electrode is inserted into the right front foot and the other into the right rear foot.

Body temperature, mean arterial pressure (MAO), ECG, and heartbeat are constantly recorded throughout the experiment. Once the critical parameters have stabilized, a 1-2 minute record is made to establish the base values. Infusion of a compound of the invention or the control substance is started once the base values ​​have been established. After a 5 minute infusion of compound or control, the suture is pulled tightly to ligate the ACL and generate ischemia in the left ventricle. Critical parameters are recorded continuously for 20 minutes after ligating the track, unless MAP reaches the critical level of 20-30 mm Hg for at least 3 minutes, in which case the recording is stopped because the animal would be declared dead and would be sacrificed. The ability of the compounds of the invention to prevent arrhythmias and maintain almost normal MAP and HR (heart beat) values ​​is rated and compared to control.

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BIOLOGICAL EXAMPLE 6

Compared to the racemic mixture, i.e., the compound of formula (I), the (S) -enantiomer, substantially free of (/?) 5 enantiomer, has a better solubility profile in a variety of pharmaceutically acceptable excipients. Thus, the (S) -enantiomer can be formulated in a smaller number of dose units than the racemic mixture. This property makes it easier to dose patients to a higher level if necessary to achieve efficacy. Examples of the difference in solubility are shown in Table 5 below:

TABLE 5

<td>Excipient</td><td>Compound of the formula (1) (racemic mixture))</td><td>(S) - enantiomer</td>

<td>Labrasol®</td><td>72.5 mg / mL</td><td>231 mg / mL</td>

<td>Propylene glycol</td><td>2.7 mg / mL</td><td>9.8 mg / mL</td>

<td>PEG 400</td><td>&lt;50 mg / mL</td><td>&gt; 55 mg / mL</td>

<td>Capryol® 90</td><td>18.1 mg / mL</td><td>96 mg / mL</td>

<td>Tween® 80</td><td>64 mg / mL</td><td>&gt; 123 mg / mL</td>

<td>Ethanol</td><td>10.0 mg / mL</td><td>36.4 mg / mL</td>

<td>Labrasol® / PEG 400 60/40</td><td>70.4 mg / mL</td><td>182 mg / mL</td>

<td>Labrasol® / Capryol®90 60/40</td><td>44.4 mg / mL</td><td>191 mg / mL</td>

<td>Labrasol® / T ranscutol® 60/40</td><td>74.2 mg / mL</td><td>186 mg / mL</td>

BIOLOGICAL EXAMPLE 7

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<img file="MX342298B_D0093.tif" />

Live trial for the treatment of pruritus

Histamine causes itching (itching) in humans. Accordingly, this assay evaluates the efficacy of the (S) -enantiomer of the invention administered topically and orally on histamine-induced itching in ICR male mice.

Animals were randomly divided into test groups, including an untreated group, a group treated with a topical composition with 8% (w / v) (S) -enantiomer, and a group treated with a 50mg oral pharmaceutical composition. / Kg of (S) -enantiomer. One day before the test, the scapular regions of the animals were shaved with barber scissors. On the day of the test the animals were habituated for 60 minutes in the test chamber composed of a transparent plastic tube placed vertically on a flat surface. After the adjustment period, the animals were removed from the plastic tube, placed in restriction and injected with histamine in the shaved scapular region. Injections were made intradermally into the skin in small injection volumes (10 pL) using a Hamilton syringe. The injected solutions consisted of histamine dissolved in saline in a concentration of 100 pg / 10 pL (or 10 mg / mL). Each mouse was injected with 10 pg of the solution. Immediately after injections. Animals were returned to test chambers and observed for a total of 50 minutes.

INDUSTRIAL ^ = »- by means of cameras placed on the test chambers. The cameras ___ were connected to a computer in which digital files were created, saved and analyzed.

The number of itching attacks over 40 minutes was rated.

An act of itching was defined as the act of lifting the hind leg and using it to scratch the scapular region, and then return it to the ground. Alternatively, if instead of placing the hind paw on the ground the mouse licked the paw, then that was also counted as an itchy attack.

In the untreated group, the animals (n = 7) were habituated in the test chamber for 60 minutes before the histamine was injected. To assess the topical (S) -enantiomer in histamine-induced pruritus, animals (n = 16 / group) were habituated in the test chamber for 30 minutes, followed by application to the shaved back region of 50 mg 6% (w / v) topical (S) -enantiomer or from a vehicle. The animals were returned to the test chamber for another 30 minutes of habituation before the histamine was injected. To assess the oral (S) enantiomer, animals (n = 8 / group) were dosed by oral gavage with 50 mg / Kg of the (S) -enantiomer or a vehicle, followed by a period of 60 minutes of habituation before the histamine injection.

Data were analyzed using GraphPad Prism 5 statistical analysis software and an unpaired t-test was used for univariate (univariate) analysis. Results are expressed as the mean ± SEM. Values ​​that reached a significance level of p &lt;0.05 were

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<img file="MX342298B_D0094.tif" />

considered statistically significant

Results

Injecting histamine into the skin caused the animals to sporadically itch 5 in attacks lasting 1-2 seconds. In the untreated group, the itching attacks started immediately after the injection and lasted, more or less, 40 minutes thereafter (see FIG. 4). The group treated with the topical (S) -enantiomer at 8% (w / v) showed a significantly reduced itching (see FIG. 5). Animals treated with the vehicle alone had a total number of itch attacks of 134.3 ± 13.31 (n = 16) while mice treated with the topical (S) -enantiomer had 89.00 ± 10.51 (n = 16) itch attacks. The difference between these groups was statistically significant with a p-value of 0.0122. The group treated with 50 mg / Kg of oral (S) -enantiomer similarly showed significantly reduced itching (see FIG. 6). Animals treated with the vehicle alone had a total of 42.88 ± 6,667 (n = 8) itch attacks, while mice treated with the (S) -enantiomer had 17.25 ± 6,310 (n = 8) itch attacks. The difference between the orally treated groups was also statistically significant with a p value of 0.0144. The results showed that the (S) -enantiomer administered orally and topically reduced itching.

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<img file="MX342298B_D0095.tif" />

BIOLOGICAL EXAMPLE 8

Human clinical trial for the treatment of primary / hereditary ritromelalgia (IEM)

Primary / Hereditary Erythromelalgia (IEM) is a rare inherited painful condition. The underlying cause of EMI may be one or more gain-of-function mutations in the Na / 1.7 voltage-gated sodium channel, which has been shown to be Inhibited by the (S) -enantlomer of the Invention.

Human patients with IEM have recurrent episodes of intense burning pain associated with redness and warming of the hands and feet, but eventually the pain becomes constant. The pain calms down on cooling but has long been resistant to pharmacological intervention. However, there are reports of voltage regulated sodium channel blockers showing moderate to outstanding relief for this condition.

A clinical trial to determine the efficacy of the (S) enantlomer of the Invention in improving or alleviating EMI can be designed to be a three-period, double-blind, multiple dose, cross-over chromosomal study to minimize the Participant dropout rate, and will take into account that enrolled patients will only be available for a 10-day study. Each patient enrolled in the study will serve as their own control, receiving the placebo and the N »DTWAL

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400 mg of the (S) -enantiomer of the invention two times a day with a chromosomal crosslinked modcrdr * ^ (w / oss-over).

BIOLOGICAL EXAMPLE 9

Clinical trial in humans for the treatment of dental pain

The purpose of this clinical trial was to compare the safety and efficacy (baseline, duration of relief, and overall efficacy) of a single 500 mg dose of the (S) -enantiomer versus a placebo dose for pain relief after extraction of a third molar.

Sixty-one subjects were enrolled in the study. The mean age of the subjects was 20.4 years and all males. Most of the subjects were Caucasian (95.1%).

Pain severity and relief was measured using an 11-point Numerical Pain Intensity Rating Scale of 15 (graded from 0 = no pain at all to 10 = worst pain imaginable) (PINRS) and a Relief Scale 5-Point Categorical Pain (REL). Subjects passed the PINRS test after surgery but before administration of the (S) -enantiomer of the invention. Efficacy variables were derived from the REL and PINRS scores, including total pain relief (TOTPAR), difference in pain intensity (PID), and summed pain intensity difference (SPID). They were evaluated at time points at 4, 6, 8 and 12 hours after administration.

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^{, Ν? Π}£? Α,&gt;_R; .F! TI) To&gt; INDUSTRIAL of the (S) -enantiomer of the invention.

However, the primary and all secondary endpoints showed a trend consistent with a clear separation between the (S) -enantiomer and placebo. These results suggest that the (S) enantiomer has analgesic properties but a statistical significance of the placebo was not obtained due to two reasons; (1) a relatively high response to placebo, and (2) the slow onset of action of the (S) -enantiomer. The dental model used is designed for and best suited for evaluation of rapid-onset drugs such as the NSAID class of anti-inflammatory agents. It was evident from this study that the (S) enantiomer of the invention does not have a fast start activity like the NSAID. However, the pain relief shown by those subjects who received the (S) -enantiomer was greater compared to the subjects who only received the placebo, enough that the efficacy in the total population showed a consistent analgesic signal in all the evaluated endpoints.

BIOLOGICAL EXAMPLE 10

Human clinical trials of the Safety of the (S) -enantiomer of the

Invention

This clinical trial was Phase 1, randomized, double-blind, placebo-controlled trial of a study in healthy subjects to assess the

100

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Safety and Pharmacokinetics of a Topically Applied Ointment Containing the (S) -enantlomer of the Invention.

The (S) -enantlomer ointment was applied daily for 21 consecutive days to determine the toxicity / local toxicity of the (S) enantlomer. Pharmacokinetics and local drug levels were also evaluated. Systemic exposure to the (S) -enantlomer after topical applications and local irritation after multiple doses of the ointment with (S) -enantiomer were evaluated. Each subject received 5 treatments on 21 consecutive days: (S) -enantlomer as ointment with 4% and 8% (w / w) (1 x 100 pL; Treatments A and B, respectively), placebo in ointment (Treatment C), 0.9% saline (1 x 100 pL; negative control; Treatment D), and 0.1% sodium lauryl sulfate (SLS) solution (1 x 100 pL; positive control; treatment E). The treatments were applied at two different sites on the upper back of each subject in a closed site (5 treatments) and in a partially closed site (the first three treatments). The location for each treatment at each site (Treatments A, B, C, D, and E in a closed site and treatments A, B, and C in a partially closed site) was randomized (ramdomised). Subjects were confined in the Clinical Investigation Facility from approximately 18 hours before the first dose on Day 1 to approximately 8 hours after the second dose (Day 2). Subjects returned each day for 19 continuous days (Days 3 to 21) for study doses and procedures.

No Serious Adverse Events (SAE) were reported or

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deaths. All Adverse Events (AE) were of minor or moderate severity, the majority of AE being related to local skin reactions due to the adhesive tape used to close the bandages. All subjects reacted to the positive control. The positive control was stopped in all subjects on Day 4 after complaints of excessive discomfort raised by the subjects. Skin irritation scores were low for all treatments administered (highest score was 3 on a scale of 0-7) indicating that (S) enantiomer ointment was well tolerated locally. No difference was seen between the cumulative irritation scores for the (S) -enantiomer 4% (w / w), the (S) -enantiomer 8% (w / w), the placebo ointments and the negative control ( 0.9% saline solution). The signs of irritation had completely resolved by Day 28 (7 days after the last dose) in most subjects.

. Electrocardiogram traces showed no significant clinical changes in pulse, quiescent resting state, or QT intervals._C of subjects, or any significant clinical changes from baseline in subjects' vital signs, physical examinations, or laboratory evaluations. Systemic exposure to (S) -enantiomer was ruled out, since plasma (S) -enantiomer concentrations were below the limit of quantification (LLOQ) (0.1 ng / mL or 100 pg / mL) in most of the samples (489 of 546 = ~ 90%). The highest observed level of (S) enantiomer in a subject during the dosing period (Day 22) was

102

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994 pg / mL. Based on the minimal local irritation and the favorable safety profile, together with the low systemic exposure of the (S) -enantiomer, it was concluded that the (S) -enantiomer of the Invention was well tolerated and safe as a topical analgesic.

BIOLOGICAL EXAMPLE 11

Clinical trials in humans for the treatment of po therpetic neuralgia

Post-Herpetic Neuralgia (PHN) is a well-established and well-recognized model for the study of neuropathic pain. Furthermore, PHN shows strong evidence of efficacy by sodium channel blockers. The following study represents a randomized, double-blind, placebo-controlled, two-treatment, two-period chromosomal cross-over study to assess the safety, tolerability, primary efficacy, and systemic exposure to the (S) -enantiomer of the invention topically administered to patients with PHN. The primary objectives are (a) to compare the safety and efficacy of an ointment containing the (S) -enantiomer with that of a placebo, in pain relief in patients with PHN, and (b) to evaluate the extent of systemic exposure of the (S) -enantiomer after topical application of the (S) -enantiomer to patients with PHN. Treatments will consist of 8% (w / w) (S) -enantiomer ointment and the corresponding placebo ointment.

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<img file="MX342298B_D0100.tif" />

The study will include the following four periods:

one. An initial filtering and washing period (up to 3 weeks);

2. A single-blind and placebo period (1 week);

3. A chromosomal cross-over treatment period consisting of 2 treatment periods each of 3 weeks separated by 2 weeks of wash / mono-blind runs (total of 8 weeks); and

Four. A safety monitoring period (2 weeks).

All US patents, US patent application publications, patent applications, foreign patents, foreign patent applications, and non-patent publications, referred to herein herein are incorporated by reference in their entirety.

Although the preceding invention has been described in some detail for ease of understanding, it will be apparent that some changes and modifications can be made within the scope of the appended claims. Accordingly, the disclosed embodiments are to be considered as illustrative and not restrictive, and the invention is not to be limited to the details disclosed herein, but may be modified within the scope and equivalents of the appended claims.

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