NZ612804B2 - Treatment for lipodystrophy - Google Patents

Abstract

Provided are pyrrole derivative compounds of the general formula (I) where the variables are as defined in the specification. An example of the compounds is (S)-?-Ethoxy-4-[2-[-methyl-S-[4-(methylthio)phenyl]-1H-pyrrol-1-yl]ethoxy] benzenepropanoic acid magnesium salt. Further provided is the use of the compounds in the treatment of lipodystrophy. of the compounds in the treatment of lipodystrophy.

Description

TREATMENT FOR LIPODYSTROPHY Field of the invention The present invention is related to the development of eutic compound for prevention and treatment of strophy. In particular the ion relates to the development of therapeutic compound for prevention and treatment of lipodystrOphy in ..

HIV —infected patients (LDHIV). Specifically, the present invention further. provides a suitable composition useful in the treatment or prevention or alleviation of the symptoms of lipodystrophy in HIV infectedpatients (LDHIV) Background of the invention 1O Lipodystrophy is a very dreadful disease and has become a major global health problem. It is a er of fat metabolism which causes lipohypertrophy, Lipoatrophy and Metabolic abnormalities. er, lipohypertrophy‘ includes the enlargement of dorsocervical fat pad nly called "buffalo hump"), expansion of the circumference of the neck by 5-10 cm, rophy Occuring in breast, central truncal adiposity resulting from abdominal visceral fat accumulation, symmetric and asymmetric lipomatoses. A rare pattern of lipoaccumulation involves formation of band like lipomatosis tissue symmetrically from the breasts, laterally to the axillae, Suprapubic fat pads (pubic lipomas) and the development of multiple angiolipornas.

Lipoatrophy includes a temporal wasting and loss of subcutaneous fat from the ‘cheeks (buccal fat pad) which produces an emaciated appearance with prominent nasolabial creases. Further subcutaneous tissue is depleted fromthe arms, shoulders, , and buttocks (peripheral wasting), with prominence of the superficial veins in these sites.

Metabolic abnormalities include augmentation in cholesterol and ceride- levels and reduced high-density lipoprotein (HDL) terol levels, n resistance, type 2 diabetes mellitus, and lactic academia.

Lipodystrophy is very commonly associated with the HIV patients who. are being treated anti-retroviral medicines. Such medicines can include HIV-I protease inhibitors (PIS), Nucleoside reverse transcriptase inhibitors (NRTIs), Non-nucleoside Reverse riptase Inhibitors (NNRTls), Fusion Inhibitors, Entry Inhibitors - CCRS co-receptor nist, HIV ase strand transfer inhibitors etc. These medicines improve the survival of the patient but also produce lipohypertrophy, rophy and other Metabolic abnormalities.

HIV-] se inhibitors (Pls) appear to be the strongest link to lipodystrophy in HIV —infected patients LDHlV as it ts maturation of sterol response element binding proteins (SREBP), which affect intracellular fatty acid and glucose metabolism and adipocyte differentiation (Mallon et al, J Infecr Dis, 2005). Furthermore, the P15 also dowu-regulate peroxisome proliferator-activated receptor gamma (PPAR 7), an ant nucleartranscription factor that is' affected by SREBPs and is necessary for adipocyte differentiation and function and fatty acid metabolism.

Other factors,’ such as duration of HIV infection, age, and gender, may also contribute to the risk of development of LDHlV. The molecular basis of LDHIV is still remains unknown and no specific therapy is available for LDHlV.

Reverse transcriptase inhibitors (nRTls) like stavudine, didanosine and zidovudine may cause mitochondrial ty by inhibiting ’ mitochondrial DNA polymerase—yin. fat and other tissues and thus interfering with respiratory chain xes. The result is impaired fatty acid oxidation and intracellular accumulation of ' cerides and lactate In addition, lipodystrophy is also observed in acute HIV infection, lending support to a direct viral role as well. Potential host risk factors include age, sex, and race or ethnicity. Lipodystrophy is more common in older patients; fat accumulation is more common in women and lipoatrophy in men; and spanic black patients appear to be at lower risk of rophy. A genetic ent is indicated by a recent analysis in AIDS Clinical s Group (ACTG), study 50055, suggesting either predisposition or protection associated with mitochondrial DNA polymorphisms.

Hulgan et al, J Infect Dis, 2008 describes that patients homozygous for C/C at the HFEl87 locus (n = 71) had a 0.6-kg and 12.5% loss oflimb fat at weeks 48 to 64, with 37 (52%) of the 7l patients diagnosed with clinical rophy. By comparison, heterozygous patients with HFEl87C/G had a 0.2—kg and 6.1% increase in limb fat, with 6 (26%) of 23 patients having clinical lipoatrophy (P < .05 for all isons).

A number of strategies for reducing central obesity have been investigated such as stopping Pl treatment but it is not effective. Changes in diet and exercise have produced improvements, but adherence to a regimen of lifestyle change is difficult for most patients. Liposuction may be applied particularly with dorsocervical fat lation, i.e., “buffalo hump”. 2012/000069 it is evident from the l 's'tudies that thiazolidinediones show no change in VAT (Pathogenesis and treatment of lipodystrophy, vol.l6, issue 4, vember,2004) I Testosterone replacement to physiologic levels reduces visceral adipose tissue (VAT), total fat, and abdominal fat and improves insulin sensitivity and lipid profile-in older, non—HIV—infected men with upper body obesity and low testosterone levels. In a recent study, 88 HIV-infected men with l obesity (waist ference >100 cm) and low testosterone levels (< 400 ng/dL) underwent ization to testosterone as a transdermal gel at a dose of )0 g daily or placebo for'24 weeks (Bhasin et al, J. Clin Endocrinol Metab, 2007). The testosterone group had statistically significant reductions in abdominal fat {—1.5% vs +43%), abdominal aneous adipose tissue (SAT) (— 7.2% vs +8.l%), trunk fat (—9.9% vs +46%), and'limb fat (~10,l°/o vs +3.l%); the latter finding is of potential concern in a tion predisposed to lipoatrophy. No statistically significant difference in change in VAT (+0.9% vs +23%) was ed, and no statistically significant ences were observed in changes in lipid levels, ' fasting blood glucose levels, insulin levels, or insulin resistance. ' hormone Like testosterone, growth (GH) has fat-oxidizing and lipolytic prOpertiesJ A substantial proportion of HIV patients with central obesity (approximately %—40°/o) have impaired GH biology, including reduced GH mass secretion, reduced response to GH releasing e (GHRH) and free fatty acids, and increased somatostatin tone, which suppresses GH. A number of recent studies have assessed GH ent in HIV patients-with" fat accumulation. in I study, 325 HIV patients with increased waist: hip ratios and increased VAT measurements received.

Although, the growth hormone (GH) and GH releasing hormone (GHRH) therapies show some promising result as they have fat—oxidizing and lipolytic properties however, there are limitations to their use. They are parenteral therapies and either expensive (rhGH) or not FDA—approved (tesamorelin). Thus far,- there is evidence of waning durability of the reduction in VAT afier their discontinuation, short-term increases in insulin resistance with rhGH, and small term reductions.

Recent research publications have shown the use of two lipid-lowering classes of drugs, statins and fibrates, antiretroviral switching strategies and use of insulin- sensitising drugs as having some beneficial effect on lipodystrOphy. However, no single therapy is able to reach desirable al end point for HIV associated lipodystrophy. 011NZPR 303903983 It is an object of the t invention to develop a compound which can overcome the above discussed drawback associated with prior art and develop a therapy for HIV associated lipodystrophy. It is an alternative object of the present invention to provide use of a compound in the manufacture of a medicament for the treatment of lipodystrophy and/or use of the compound for the preparation of a ment for the treatment of lipohypertrophy or lipoatrophy or metabolic ality and/or a ceutical ition for the treatment of lipodystrophy and/or use of a pharmaceutical composition in the manufacture a medicament for the treatment of HIV associated lipodystrophy and/or use of a pharmaceutical composition in the manufacture of a medicament for the treatment of lipohypertrophy or lipoatrophy or metabolic abnormality and/or use of a pharmaceutical composition in the manufacture of a medicament which reduces the concentration of triglyceride, very low density otein, Apo B level and/or use of a pharmaceutical composition in the manufacture of a medicament which increases high density lipoprotein, Apo A1 level and/or a novel compound and/or use of a compound in the manufacture of a medicament which reduces the concentration of triglyceride, very low density lipoprotein, Apo B level and/or use of a compound in the manufacture of a medicament which increases high density lipoprotein, Apo A1 level and/or a pharmaceutical composition comprising the nd. It is a further or alternative object of the present invention to provide the public with a useful choice.

Hypolipidemic agents which are PPAR modulators have been disclosed in WO 91/19702, WO 94/01420, WO 94/13650, WO 95/03038, WO 94, WO 96/04260, WO 96/04261, WO 96/33998, WO 42, WO 79, WO 98/28534, WO 99/08501, WO 99/16758, WO 99/19313, WO99/20614, WO 00/23417, WO 00/23445, WO 00/23451, WO 01/53257.

WO 03009841 discloses compounds of the following general formula R1 R5 Y N (CH2)n W Ar ZR 8 R3 XR7 VIA510011NZPR 303903983 These compounds are reported to be hypolipidaemic agents. This document also discloses sodium and calcium salts of some of the nds disclosed therein. r, the sodium salts of the compounds of the present invention was difficult to isolate due to rapid ation while the Calcium salt was poorly absorbed limiting its efficacy and possibility of further development. Further, the calcium salt was also found to degrade on long term storage. It has surprisingly now been found that certain compounds and their selected salts are effective in the treatment of lipohypertrophy, lipoatrophy and metabolic abnormalities in HIV patients. ments of the invention In an embodiment the t invention provides a compound of formula (I) suitable for the ent and prevention of strophy.

In an embodiment, the conditions associated with lipodystrophy includes the symptoms of lipohypertrophy, lipoatrophy and other metabolic abnormalities.

In another embodiment, the present invention provides a compound of formula (I) for the treatment and prevention or alleviation of symptoms of lipohypertrophy, lipoatrophy and metabolic abnormalities in HIV t.

In yet another embodiment the present invention es the administration of compound of formula (I) and their pharmaceutically acceptable salts alone or in combination with other suitable agents as therapeutic agent for the treatment and prevention alleviation of symptoms of lipodystrophy.

In yet another embodiment the present invention provides a suitable ition comprising the compound of formula (I) or their suitable pharmaceutical compositions suitable for the treatment and prevention alleviation of symptoms of strophy.

In another embodiment, the present invention provides for certain pharmaceutical salts of compound of formula (I).

VIA510011NZPR 303903983 In a first aspect, the t invention provides use of a compound of formula O M+ wherein ‘R’ is selected from hydroxy, hydroxyalkyl, acyl, alkoxy, alkylthio, kyl, aryloxy, arylthio and M+ represents suitable metal cation selected from Mg+2 in the manufacture of a medicament for the treatment of lipodystrophy.

In a second aspect, the present invention provides use of the compound of a (I) as defined in the first aspect in the manufacture of a medicament for the treatment of lipohypertrophy or lipoatrophy or metabolic abnormality.

In a third aspect, the present invention provides use of a compound of formula (I) as defined in the first aspect in the manufacture of a ment which reduces the concentration of triglyceride, very low density lipoprotein, Apo B level.

In a fourth aspect, the present ion provides use of a compound of formula (I) as defined in the first aspect in the manufacture of a medicament which ses high density lipoprotein, Apo A1 level.

In a fifth aspect, the present ion provides a compound of formula (Ia) O M+ (Ia) VIA510011NZPR 303903983 wherein ‘R’ is ed from hydroxy, hydroxyalkyl, acyl, alkoxy, alkylthio, thioalkyl, aryloxy, arylthio and M+ represents suitable metal cation Mg+2.

In a sixth aspect, the present invention provides a pharmaceutical composition for the treatment of lipodystophy comprising: a) a compound of formula (Ia) as defined in the fifth ; b) a suitable stabilizer; c) a suitable buffering agent; d) optionally, with one or more pharmaceutically acceptable excipients.

In a seventh aspect, the present invention provides use of the pharmaceutical ition as defined in the sixth aspect in the manufacture of a medicament for the treatment of HIV associated lipodystrophy.

In an eighth , the present invention provides use of the pharmaceutical composition as d in the sixth aspect in the manufacture of a medicament for the treatment of pertrophy or lipoatrophy or metabolic abnormality.

VIA510011NZPR 303903983 In a ninth aspect, the present invention provides use of the pharmaceutical composition as d in the sixth aspect in the manufacture of a medicament which reduces the concentration of triglyceride, very low density lipoprotein, Apo B level.30.

In a tenth aspect, the present invention provides use of the ceutical composition as defined in the sixth aspect in the manufacture of a medicament which increases high density lipoprotein, Apo A1 level.

In an eleventh aspect, the present invention provides a pharmaceutical composition comprising a compound of formula (I) as d in the fifth .

In a twelfth aspect, the present invention provides a pharmaceutical composition comprising compound of formula (Ia) as defined in the fifth aspect for the treatment of lipodystrophy.

Summary of the invention The t invention provides a compound of formula (I) and their pharmaceutically acceptable salts for the tion and treatment or alleviation of symptoms of lipodystrophy. The present invention es a nd of formula (I) and their pharmaceutically acceptable salts for the prevention and ent or alleviation of symptoms of lipodystrophy caused either because of HIV infection or due to treatment with anti-retrovirals. Such anti-retrovirals can include HIV-1 protease inhibitors (PIs), Nucleoside reverse transcriptase inhibitors (NRTIs), Non-nucleoside Reverse Transcriptase tors (NNRTIs), Fusion Inhibitors, Entry Inhibitors - CCR5 co-receptor antagonist, HIV integrase strand transfer inhibitors etc. or combination y involving one or more anti-retrovirals. The compound of formula (I) neutralizes lipohypertrophy, lipoatrophy and metabolic abnormalities in HIV patient. Moreover, the present invention also provides a suitable composition comprising compound of formula (I) useful in the treatment or prevention or alleviation of the symptoms of lipodystrophy in HIV infected patients (LDHIV).

In a further embodiment are disclosed certain new salts corresponding to the compound of formula (I) wherein M represents K or Mg.

VIA510011NZPR 303903983 Description of the Invention The present invention bes compound of formula (I) which is suitable for the treatment of lipodystrophy or HIV associated lipodystrophy.

O M+ PCT/1N2012/000069 wherein ‘R’ is selected from‘hydroxy, hydroxyalkyl, acyl, alkoxy, alkylthio, thioalkyl, aryloxy, arylthio and M+ represents suitable metal cations such as Na‘, K, Ca”, Mg? and the like. , In a preferred ment, ‘R’ represents thioalkyl, alkoxy or hydroxyalkyl ‘R’ ents —SCl-l3 or -OCH3 group. group; In a still red embodiment, ln an embodiment is provided suitable pharmaceutical composition for the treatment of lipodystrophy or HIV associated lipodystrophy sing the compound of formula (I). The pharmaceutical 'composition of the present invention comprises compound of formula (I) along with suitable excipients as defined after for the treatment of strophy or. HIV associated lipodystrophy.

In another embodiment, the present ion provides a method of treating a subject suffering from lipodystrophy or HIV associated lipodystrophy which comprises treatment of a patient in need of such therapy, with compound of formula (1) or suitable pharmaceutical compositions containing them.

In a further embodiment the present invention provides use of the compound of formula (I) or their suitable pharmaceutical compositions for the treatment of lipodystrophy or HIV associated lipodystrophy.

In an ment the present invention provides certain new salts of compound of formula (la) O ' CH3 _ 0 + / /\/0 N r (1a) wherein ‘R’ is selected from hydroxy, hydroxyalkyl, acyl, alkoxy, alkylthio, thioalkyl, aryloxy, arylthio and M+ ents le metal cations selected from K”, Mg”.

In a preferred embodiment, ‘R’ represents thioalkyl and alkoxy or hydroxyalkyl ‘R’ represents —SCH3 or -OCH3 group. group; In a still preferred embodiment, In another preferred embodiment, M+ represents Mg”. is selected from 1 mg The effective amount of the said compound of formula (I) 1 mg to 250 mg and more preferably 4 mg to 50 mg. The to 500 mg preferably compound of formula (I) or its suitable salts is administrated orally, intravenously, parentally in the subject whois in need of treatment.

In an embodiment the compound of formula (1) is useful for the treatment or In a preferred embodiment prevention or ation of the symptoms of lipodystrophy. the compound of formula (I) is useful in the treatment or prevention or alleviation of embodiment the Lipodystrophy the symptoms ofHIV associated lipodystrophy. In such and metabolic is a disorder of fat metabolism which causes lipohypertrophy lipoatrophy ' abnormalities. alleviate at In an embodiment the compound of formula (1) cure or prevent or limited to, acting as an agent least one symptoms of lipodystrophy ing, but not for lowering &/or control blood glucose levels, an agent used to control lipid , agent used to lower control cholesterol, an e.g., as an antioxidant, an appetite suppressing agent, an anti-obesity agent, a‘probiotic or an anti-inflammatory agent. alleviate at another embodiment the the compound-of a (I) cure or prevent or limited to triglyceride level, least one symptoms of strophy including, but not VLDL level and Apo B level in serum. In r embodiment the compound of least one of the condition a (I) cure or prevent of lipodystrophy by improving at ed from HDL level, Apo A] level, HOMA of beta cell function derived from c- peptide. suitable pharmaceutical In an embodiment the present invention also provides a composition of compounds of formula (I) or their derivative. The pharmaceutical ition of the present inVention ially comprises of: - the pharmaceutically active substance; - 'a suitable buffering agent; - a suitable izer; - optionally with one or more-pharmaceutically acceptable excipicnts.

The suitable stabilizers used in pharmaceutical composition are selected from Potassium chloride, Sodium stearyl fumarate and Polacrilin potassium, ably selected from selected from Sodium stearyl fumarate. The suitable buffering agent are W0 2012/104869 sodium e, ammonia solution, ammonium carbonate, sodium borate, adipic Acid, glycine, monosodium glutamate and preferably selected from ammonia solution.

The ceutically acceptable excipients are selected at least one from carriers, s, antiOxidant agents, disintegrating agents, wetting agents, lubricating agents, ing agents, surface active agents, and the like.

Diluents include, but are not limited to lactose monohydrate, e, polymethacrylates selected from Eudragit, potassium chloride, sulfobutylether b- cyclodextrin, sodium chloride, spray dried lactose, and preferably sulfobutyl ether b- cyclodextrin. Carriers include, but are not limited‘to lactose, white sugar, sodium chloride, glucose, urea, , calcium carbonate and kaolin, crystalline cellulose, and silicic acid. Binders include, but are not limited to carbomers selected from ol, gellan, gum Arabic, hydrogenated vegetable oil, polymethacrylates selected from Eudragit, xanthan, lactose and Zeiri. Antioxidant agents include, but are not limited to, Hypophosphorous acid, Sodium formaldehyde, sodium forrnaldehylde sulfoxylate, sulfur dioxide, tartaric acid, thymol and methionine. egrating agents include, but are not limited to, bicarbonate salt, chitin, gellan gum, illin potassium and Docusate Sodium. Wetting agents include, but are not limited to, Glycerin, lactose, te Sodium and Glycine, Lubricating agents used e, but are not limited to, Glycerin behenate, hydrogenated vegetable oil, sodium stearyl fumarate and Myristic Acid. Chelating‘ agents include, but are not limited to, Maltol and Pentetic Acid.

‘ Surface active agents include but are not limited to, Nonionic surfactant selected from alkyl polyglucosides, cocamide DEA, cocamide MBA, cocamide TEA, decyl maltoside and octyl glucoside; Anionic surfactant selected from arachidic acid and arachidonic acid; Cationic surfactant selected from cetyl trimethylammonium bromide '25 and cetylpyridinium chloride. ln an embodiment the formulation is useful for the treatment or prevention or alleviation of the ms of lipodystrophy. In a preferred embodiment the said formulation is useful in the treatment or tion or alleviation of the symptoms of HIV ated lipodystrophy.

Lipodystrophy is a disorder of fat lism which causes lipohypertrophy, lipoatrophy and lic abnormalities. Moreover, lipohypertrophy includes the. enlargement of dorsocervical fat pad (commonly called "buffalo hump"), expansion of 4 the circumference of the neck by 5-10 cm, hypertrophy occurs in breast, Central truncal adiposity results from nal visCeral fat accumulation, symmetric and asymmetric lipomatoses. A rare pattern of lipoaccumulation involving‘bandlike lipomatosis tissue symmetrically from the breasts, laterally to the axillae, suprapubic fat pads (pubic lipomas) and the development of multiple angiolipomas.

Lipoatrophy includes a temporal wasting and loss of subcutaneous fat from the cheeks (buccal fat-pad) produces an‘ emaciated appearance with prominent nasolabial creases, subcutaneous tissue is depleted from the arms, shoulders, , and buttocks (peripheral wasting), with prominence of the superficial veins in these sites.

Metabolic abnormalities include tation in cholesterol and triglyceride levels and reduced high-density lipoprotein (HDL) cholesterol levels. n resistance, type 2 diabetes mellitus, and lactic academia.

The compounds of the t invention due to their beneficial effect on lipodystrophy, will have beneficial effect on Body fat redistribution ( Lioatrophy or Hypertrophy or al distribution), Dyslipidemia, Glucose tatis, Pro- inflammatory conditions, impact on morbidity and mortality, impact on y of life, impact on patient’s reported outcomes like self perception etc , Moreover, the precise mechanisms underlying this syndrome are not well understood, several hypotheses based on in vitro and human studies may explain the pathogenesis of the changes. Some experts presently e that HIV type 1 (HIV-1) protease inhibitors (P15) and nucleoside reverse transcriptase inhibitors ), eSpecially stavudine and dine, are implicated as follows: (i) decreased productiOn of retinoic acid and triglyceride uptake: Pls have-a high affinity for the catalytic site of HIV-1 prOtease, which shares a 60% ce homology with 2 proteins involved in lipid metabolism, cytoplasmic retinoic acid— binding protein type 1 -l) and. low-density lipoprotein receptor—related protein (LDLR-RP). Inhibition of CRABP-l impairs the production of retinoic acid, leading to decreased fat storage and yte apoptosis with the subsequent release of lipids into the circulation. Inhibition of LDLR-RP results in hyperlipidemia secondary to the failure of hepatic and endothelial l of chylomicrons and triglycerides from the circulation. (ii) inhibition of mitochondrial DNA (mtDNA) polymerase gamma: NRTls inhibit mtDNA polymerase gamma, leading to mtDNA depletion, respiratory chain dysfunction, and reduced energy tion, which, in turn, causes insulin resistance and secondary'dyslipidemia. Interestingly, mtDNA is depleted only at normal oxygen levels—hypoxic adipocytes do not take up triglycerides and are resistant to mtDNA-induced damage, except after treatment with NRTIs. (iii) inhibition of lipid metabolism: Some Pls, particularly ritonavir, inhibit cytochrome P450 3A, a key enzyme in lipid metabolism. . (iv) prevention of the development of adipocytes: Saquinavir, ritonavir, and nelfrnavir (all Pls) directly inhibit the development of ytes from stem cells and se the metabolic destruction of fat in existing ytes.

In an embodiment the compound of formula (I) or pharmaceutical composition ning the compound of formula (I) cure or prevent or alleviate at least one symptoms of lipodystrophy including, but not limited to, acting as an agent for lowering &/or an agent used to control blood glucose levels, an agent used to control lipid levels, e.g., as ,an agent used to lower control cholesterol,'an antioxidant, an appetite suppressing agent, an anti-obesity agent, an otic/ tic or an anti— v inflammatory agent.. In another embodiment the pharmaceutical composition cure or prevent or alleviate at least one symptoms of lipodystrophy ing, but not limited to triglyceride. level, VLDL level and Apo B level in serum. In another ment the pharmaceutical composition cure or prevent of lipodystrophy by improving at least one of the condition selected from HDL level, Apo A] level, HOMA of beta cell function ' derived from c-peptide.

Invanother embodiment the compounds according to Formula (I) can be used alone or in combination e.g., as an adjunct therapy, with at least one other therapeutic agent. Compound ing to formula (I) can be co-administered with a eutic agent used to reduce one or more of the symptoms of lipodystrophy including, but not limited to, an agent used to control blood glucose levels, an agent used to control lipid levels, e.g., an agent used to lower l terol, an antioxidant, an appetite suppressing agent, an besity agent an antibiotic/probiotic or an anti-inflammatory agent. such combination treatment may be adjunct to anti-retroviral therapy. In a preferred embodiment the compound of formula (I) administrated alone or in combination for the treatment of Iipohypertrophy, lipoatrophy and Metabolic' abnormalities in HIV patient. 2012/000069 The compound ofthe present invention when M+ represents K, Mg can be prepared by the ses disclosed herein below along with suitable modifications known to a d person.

Example] Preparation of (S)-a-Ethoxy[2-[-methyl[4-(methylthio)phenyl]-lH-pyrrol-l- yl]ethoxy]benz_ene-propanoic acid ethyl ester In a dry,.5 L round bottom flask 2.1 L toluene was taken under nitrogen. To this 366.] g ethyl (S)- a-Z-ethoxy(4—hydroxyphenyl)propionatc was added at room ature.

The reaction mixture was stirred under heating, using Dean-stark apparatus, to remove water azeotropically. The reaction mixture was cooled ”to 50 0C. To this was added 319 g anhydrous potassium carbonate and stirred at 90-92 °C for 1 hr. Cooled to 65 0C, and added 5.00 g 2-(2-methyl(4-(methylthio)phenyl)—lH-pyrrol-l-yl)ethyl methanesulfonate and 22 g tetra butyl ammonium e. on mixture was heated to 87-92‘°C and stirred for 46 hrs. Cooled to 70-75 °C, added 1.5 L toluene, charcoalised using 75 g charcoal and cooled to room temperature. Filtrate washed with alkaline solution, washed with water, dried over sodium sulfate and concentrated under vacuum to obtain (S)-a—Ethoxy[2-[-methyl-S-[4-(methylthio)phe‘nyl]-1H-pyrrol-I- yl]ethoxy]benzene-prepanoic acid ethyl ester.

Yield: 650 g, HPLC purity: 84.10 %; % Yield 76.0 %.

Example 2 Preparation of (S)—a-Ethoxy[2-[-methyl-S-[4-(methylthio)phenyl]-1H-pyrrol-l- yl]ethoxy] benzenepropanoic acid magnesium salt.

In a dry, 250 mL round bottom flask 80 mL methanol was taken. To this 20 g (S)-a-ethoxy[2—[-methyl-5—[4-(methylthio)phenyl]- 1 H-pyrroi-l hoxy]benzene— propanoic acid ethyl ester was added at room temperature, under nitrogen. To this 1.89 g sodium hydroxide dissolved in 20 mL water was added and ed at room temperature for 3 hours to te hydrolysis. SOlvent was removed under reduced pressure. 150 mL water was added to concentrate the material. Impurity was removed by solvent washing. To aqueous layer was added 5 g magnesium acetate tetra hydrate (dissolved in 20 mL water) and stirred with for 15 min. Sticky material was extracted with dichloromethane and subsequently add n-heptane to precipitate (S)-a-ethoxy WO 04869 [2-[-methyl-S-[4-(methylthio)phenyl]-lH-pyrrol-l -yl]ethoxy]benzenepropanoic . acid magnesium salt. Solid was filtered, and dried.

Yield: 10.3 g; HPLC Purity: 98.32 %; Chiral purity: 97.64 %. the similar to those described in Examples 1 & 2 the Following process following batches of )-0t-Ethoxy[2-[-methyl[4-(methylthio)phenyl]-lH-pyrrol-l- yl]ethoxy] epropanoic acid magnesium salt were prepared. -_--“— -—_-—-- II M... — Similar reaction carried out using‘Magnesium chloride -_m--5 98.53 % Example 7 6.5 g' 79.25 % _-99.32 % Similar reaction carried out using Magnesium sulfate Example 8 82.91 % The present invention further discloses use of said nd of formula (I) or ”their suitable pharmaceutical itions for the treatment of lipohypertrophy, lipoatrophy and metabolic alities in HIV patient.

Example 9 (S)-a-Ethoxy[2-[-methyl—S-[4-(methylthio)phenyl]- l H—pyrrolyl]ethoxy] benzenepropanoic acid potassium salt. this 10 In a dry, 250 m1. round bottom flask 72 mL ethyl e was taken. To g. )o.-l-phenylethylamine salt of (S)— a-ethoxy[2—[—methyl-5—[4-(methylthio) phenyl]—lH-pyrrol-l-yl]ethoxy]benzene -propanoic acid was added at room acid (water temperature and subsequently 50 mL water and 4.8 mL dilute hydrochloric 1: 135% HCl) was added and stirred at room temperature till solid was dissolved.

Layer was separated and organic layer was washed with water, dried over sodium this was added 50 mL sulfate and solvent removed. 9.2 g oily mass obtained. To methanol and stirred under nitrogen. To this was added 1.81 g potassium xide was stirred at room temperature for 15 min. Solvent removed and added n-Hexane. under vacuum.

Again n—hexane was removed and added methanol. Solvent removed HygrOscopicmaterial obtained. Dried it under vacuum to get (S)- a-ethoxy[2-[- methyl[4-(methylthio)phenyl]-‘ l H-pyrrol- l -yl]ethoxy]benzenepropanoic acid potassium salt.

Yield- 7.6 g, (92.77 %), 'HPLC Purity 98.60 %, Chiral purity 99.56 % Example 10’ Title of Study: A Prospective, Multi-Centric, Open-Label, Single Arm Study to Evaluate the Safety and Efficacy of 4 mg of compound of formula (I) in Hypertriglyceridemia in HIV Associated Lipodystrophy.

Objectives: The objective of this study was to evaluate the safetyland efficacy of 4 mg of compound of formula (I) in hypertriglyceridemia in HIV associated lipodystrophy.

Methodology: This was a prOSpective, multi-centric, open-label, single‘ann study to evaluate the safety and y of 4 mg of compound of formula (l) in riglyceridemia in HIV associated lipodystrophy.

After obtaining informed n consent, subjects with hypertriglyceridemia in HIV associated strophy, on ent ‘with HAART for at least 18 months and satisfying the ion and exclusion criteria were enrolled in the study. The subjects received 4 mg of compound of formula (I) tablet orally, once dailyfor a period of 12 weeks. During this 12-week program, safety parameters were assessed at weeks 2, 6, and 12 and the efficacy was evaluated at week 6 and 12.

Number of patients: Planned: 50, Analyzed: 50 Test product: Compound of formula (1) Dose 4 mg Duration of ent: l2 weeks Mode of administration: Oral .

Batch number: 8 Criteria for evaluation: Efficacy: The primary efficacy endpoint was to assess the percent change in TO levels from baseline to Week 6 and Week 12. The secondary efficacy endpoint was the assessment of LDL, VLDL, HDL, Non HDL terol, Total cholesterol, Apo A1, Apo B, and C—peptide and fasting insulin for HOMA beta and HOMA IR.

Safety: W0 2012/104869 Clinical examination and recording of adverse events (AEs) was done on all . Electrocardiogram was recorded at screening visit and at Week 12. Urine pregnancy test was conducted at screening visit Haematological examination included haemoglobin, haematocrit, red blood cell (RBC) count, white blood cell (WBC) count with ential (neutrophils, cytes, monocytes, eosinophils and basophils) and et count. I Biochemistry tests ed AST, ALT, ALP, total bilirubin, serum proteins, total albumin and globulin, y- GTT, BUN, Serum creatinine, serum uric acid, CPK, and urine R/Ms (including microalbumin'uria and ketonuria).

All laboratory parameters were evaluated at ent visit (Week 0) and at Weeks 2, 6, and 12.

Statistical methods For the efficacy endpoints, treatment effect was ted using an analysis of- ce (ANOVA) model with factors for baseline and treatment. Treatment effects were estimated using the least-square means (LSM) and 95% confidence als (Cls) from the ANOVA model. Statistical significance was defined as a two-sided p-value <0.05. All other secondary endpoints were analyzed using appropriate statistical methods.

For safety analysis the frequency tabulations of abnormal physical examination and abnormal clinical tory parameters were presented for each visit. Summary statistics for al laboratory parameters and vital signs were presented for each visit.

A list of concomitant medications taken during the study period was summarised.

Adverse events were coded using the Medical Dictionary for Regulatory Activities (MedDRA) (Version 14). Adverse events and SAEs were summarized overall, by system organvclass (SOC) and by MedDRA preferred term for treatment emergent adverse events (TEAEs). All AEs, including those g before or after treatment was included in the listings. Separate listings were provided for SAEs and AEs leading to discontinuation from the study.

STUDY DESIGN This was a safety and efficacy study to evaluate 4 mg of compound of formula (I) in hypertriglyceridemia in HIV associated lipodystrophy. This was exploratory proof of concept study designed to assess the proof of safety and efficacy in intended The results of compound of formula (I) from phase ll studies in , population.

WO 04869 Dyslipidemia subjects demonstrated that compound of formula (I) 4 .mg is well tolerated and effective at once daily dosing. Phase l study demonstrated food significantly affects absorption of compound of formula (I), so drug was recommended to be consumed preferably in fasting ion. Based upon these observations 4 mg once daily in fasted condition was selected for present study SELECTION OF STUDY POPULATION Inclusion Criteria Subjects who satisfied all of the following criteria were eligible for enrolment in the study: 1. Males and females aged 18- 65 years. 2. Confirmed sis of HIV] and on HAART for at least 18 months. 3. On stable ART n for at least 8 weeks prior to inclusionin the study and ART n not expected to changein next 3 months. 4. Subjects clinically diagnosed as HIV lipodystrophy (at least 1 moderate or Severe lipodystrOphy e identified by doctor and patient, except isolated abdominal obesity) . Triglycerides >200 to 500 mg %. 6. CD4 count of>50/mm3 ‘ 7.‘ Subject who had given informed t for participation in this trial.

TREATMENTS Treatments Administered The study had a single arm. Subjects ed 4 mg of compound of formula (I) orally once daily in the morning before breakfast, for a period of 12 weeks.

Identity of lnvestigational Product(s) Compound of formula (1) is .divalent magnesium salt of carboxylic acid in the form of white, amorphous powder, which is freely soluble in dimethyl sulfoxide, dichloromethane, ly soluble in methanol and insoluble in water. The drug was supplied as uncoated tablets of 4 mg of the active ingredient.

Supply from batch no EMK328 was used during the study. The study drug was manufactured and packaged in cGMP facility.

WO 04869 2012/000069 Primary Efficacy Variable(s) The primary efficacy endpoint was to determine the percent change in T6 levels from baseline to Week 6 and Week 12.

Secondary Efficacy Variables The secondary efficacy endpoint was to determine the percent change in LDL, VLDL, HDL, total terol, non-HDL Cholesterol (measured value), Apo Al, and Apo B, C-peptide and g insulin for HOMA beta and HOMA IR levels from baseline to Week 6 and Week 12. , STATISTICAL METHODS PLANNED IN THE PROTOCOL AND DETERMINATION OF SAMPLE SIZE Statistical and Analytical Plans The demographic and baseline characteristics were summarized for nd . of formula.(I).4 mg treatment arm. For continuous measurements such as age, the mean, median, standard ion" (SD) and range were tabulated. Eor categorical measurements such as gender, the frequencies were ed.

Efficacy Analyses: The primary efficacy variable was the reduction in T0 at Week 6 and Week 12 of the treatment period compared with "baseline. The change from baseline was detennined as the difference between the means for the treatment period (Weeks