TREATMENT FOR LIPODYSTROPHY

Abstract

The present invention provides a therapeutic compound of formula (I) and their pharmaceutically acceptable salts for the prevention and treatment of lipodystrophy caused because of HIV infection or combination therapy of HIV-1 protease inhibitors (PIs) and/or reverse transcriptase inhibitors (nRTIs) by neutralizing lipohypertrophy, lipoatrophy and metabolic abnormalities in HIV patient.

Claims

1-32. (canceled)

33. A pharmaceutical composition comprising: a compound of formula (I) ##STR00004## wherein the compound of formula (I) is present in an amount between about 1 mg to about 500 mg; and a pharmaceutically acceptable excipient.

34. The pharmaceutical composition of claim 33, wherein the compound of formula (I) is present in an amount between about 1 mg to about 250 mg.

35. The pharmaceutical composition of claim 33, wherein the compound of formula (I) is present in an amount between about 4 mg to about 50 mg.

36. The pharmaceutical composition of claim 33, wherein the pharmaceutical composition is suitable to be administered orally, intravenously, or parenterally.

37. A pharmaceutical composition comprising: a) a compound of formula (I) ##STR00005## wherein the compound of formula (I) is present in an amount between about 1 mg to about 500 mg; b) a buffering agent; c) a stabilizer; d) optionally a second therapeutic agent; and e) optionally one or more pharmaceutically acceptable excipients.

38. The pharmaceutical composition of claim 37, wherein the compound of formula (I) is present in an amount between about 1 mg to about 250 mg.

39. The pharmaceutical composition of claim 37, wherein the compound of formula (I) is present in an amount between about 4 mg to about 50 mg.

40. The pharmaceutical composition of claim 37, wherein the buffering agent is selected from the group consisting of sodium acetate, ammonia solution, ammonium carbonate, sodium borate, adipic acid, glycine, and monosodium glutamate.

41. The pharmaceutical composition of claim 37, wherein the stabilizer is selected from the group consisting of polacrilin potassium, potassium chloride, and sodium stearyl fumarate.

42. The pharmaceutical composition of claim 37, wherein the one or more pharmaceutically acceptable excipients is selected from the group consisting of a carrier, a binder, an antioxidant agent, a disintegrating agent, a wetting agent, a lubricating agent, a chelating agent, and a surface active agent.

43. The pharmaceutical composition of claim 37, wherein the pharmaceutical composition is suitable to be administered orally, intravenously, or parenterally.

44. A pharmaceutical composition comprising: i) a compound of formula (I) ##STR00006## wherein the compound of formula (I) is present in an amount between about 1 mg to about 500 mg; and ii) a second therapeutic agent, wherein the second therapeutic agent is selected from the group consisting of an agent used to control blood glucose levels, an agent used to control lipid levels, an antioxidant, an appetite suppressing agent, an anti-obesity agent, an antibiotic, a probiotic, and an anti-inflammatory agent.

45. The pharmaceutical composition of claim 44, wherein the compound of formula (I) is present in an amount between about 1 mg to about 250 mg.

46. The pharmaceutical composition of claim 44, wherein the compound of formula (I) is present in an amount between about 4 mg to about 50 mg.

47. The pharmaceutical composition of claim 44, wherein the pharmaceutical composition is suitable to be administered orally, intravenously, or parenterally.

Description

DESCRIPTION OF THE INVENTION

[0028] The present invention describes compound of formula (I) which is suitable for the treatment of lipodystrophy or HIV associated lipodystrophy.

##STR00002##

wherein R is selected from hydroxy, hydroxyalkyl, acyl, alkoxy, alkylthio, thioalkyl, aryloxy, arylthio and M.sup.+ represents suitable metal cations such as Na.sup.+, K.sup.+, Ca.sup.+2, Mg.sup.+2, and the like.

[0029] In a preferred embodiment, R represents thioalkyl, alkoxy or hydroxyalkyl group; In a still preferred embodiment, R represents SCH.sub.3 or OCH.sub.3 group.

[0030] In an embodiment is provided suitable pharmaceutical composition for the treatment of lipodystrophy or HIV associated lipodystrophy comprising the compound of formula (I). The pharmaceutical-composition of the present invention comprises compound of formula (I) along with suitable excipients as defined hereinafter for the treatment of lipodystrophy or HIV associated lipodystrophy.

[0031] In another embodiment, the present invention 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 (I) or suitable pharmaceutical compositions containing them.

[0032] 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.

[0033] In an embodiment the present invention provides certain new salts of compound of formula (Ia)

##STR00003##

wherein R is selected from hydroxy, hydroxyalkyl, acyl, alkoxy, alkylthio, thioalkyl, aryloxy, arylthio and M.sup.+ represents suitable metal cations selected from K.sup.+, Mg.sup.+2.

[0034] In a preferred embodiment, R represents thioalkyl and alkoxy or hydroxyalkyl group; In a still preferred embodiment, R represents SCH.sub.3 or OCH.sub.3 group. In another preferred embodiment, M.sup.+ represents Mg.sup.+2.

[0035] The effective amount of the said compound of formula (I) is selected from 1 mg to 500 mg preferably 1 mg to 250 mg and more preferably 4 mg to 50 mg. The compound of formula (I) or its suitable salts is administrated orally, intravenously, parentally in the subject who is in need of treatment.

[0036] In an embodiment the compound of formula (I) is useful for the treatment or prevention or alleviation of the symptoms of lipodystrophy. In a preferred embodiment the compound of formula (I) is useful in the treatment or prevention or alleviation of the symptoms of HIV associated lipodystrophy. In such embodiment the Lipodystrophy is a disorder of fat metabolism which causes lipohypertrophy lipoatrophy and metabolic abnormalities.

[0037] In an embodiment 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 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, a probiotic or an anti-inflammatory agent. In another embodiment the compound of formula (I) cure or prevent or alleviate at least one symptoms of lipodystrophy including, but not limited to triglyceride level, VLDL level and Apo B level in serum. In another embodiment the compound of formula (I) cure or prevent of lipodystrophy by improving at least one of the condition selected from HDL level, Apo A1 level, HOMA of beta cell function derived from c-peptide.

[0038] In an embodiment the present invention also provides a suitable pharmaceutical composition of compounds of formula (I) or their derivative. The pharmaceutical composition of the present invention essentially comprises of:

[0039] the pharmaceutically active substance;

[0040] a suitable buffering agent;

[0041] a suitable stabilizer;

[0042] optionally with one or more pharmaceutically acceptable excipients.

[0043] The suitable stabilizers used in pharmaceutical composition are selected from Polacrilin potassium, Potassium chloride, Sodium stearyl fumarate and preferably selected from Sodium stearyl fumarate. The suitable buffering agent are selected from sodium acetate, ammonia solution, ammonium carbonate, sodium borate, adipic Acid, glycine, monosodium glutamate and preferably selected from ammonia solution.

[0044] The pharmaceutically acceptable excipients are selected at least one from carriers, binders, antioxidant agents, disintegrating agents, wetting agents, lubricating agents, chelating agents, surface active agents, and the like.

[0045] Diluents include, but are not limited to lactose monohydrate, lactose, 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, starch, calcium carbonate and kaolin, crystalline cellulose, and silicic acid. Binders include, but are not limited to carbomers selected from carbopol, gellan, gum Arabic, hydrogenated vegetable oil, polymethacrylates selected from Eudragit, xanthan, lactose and Zein. Antioxidant agents include, but are not limited to, Hypophosphorous acid, Sodium formaldehyde, sodium formaldehylde sulfoxylate, sulfur dioxide, tartaric acid, thymol and methionine. Disintegrating agents include, but are not limited to, bicarbonate salt, chitin, gellan gum, polacrillin potassium and Docusate Sodium. Wetting agents include, but are not limited to, Glycerin, lactose, Docusate Sodium and Glycine, Lubricating agents used include, 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 and cetylpyridinium chloride.

[0046] In an embodiment the formulation is useful for the treatment or prevention or alleviation of the symptoms of lipodystrophy. In a preferred embodiment the said formulation is useful in the treatment or prevention or alleviation of the symptoms of HIV associated lipodystrophy.

[0047] Lipodystrophy is a disorder of fat metabolism which causes lipohypertrophy, lipoatrophy and metabolic abnormalities. Moreover, lipohypertrophy includes the enlargement of dorsocervical fat pad (commonly called buffalo hump), expansion of the circumference of the neck by 5-10 cm, hypertrophy occurs in breast, Central truncal adiposity results from abdominal 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.

[0048] 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, thighs, and buttocks (peripheral wasting), with prominence of the superficial veins in these sites.

[0049] Metabolic abnormalities include augmentation in cholesterol and triglyceride levels and reduced high-density lipoprotein (HDL) cholesterol levels, Insulin resistance, type 2 diabetes mellitus, and lactic academia.

[0050] The compounds of the present invention due to their beneficial effect on lipodystrophy, will have beneficial effect on Body fat redistribution (Lioatrophy or Hypertrophy or abnormal distribution), Dyslipidemia, Glucose homeostatis, Pro-inflammatory conditions, impact on morbidity and mortality, impact on quality of life, impact on patient's reported outcomes like self perception etc

[0051] 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 believe that HIV type 1 (HIV-1) protease inhibitors (PIs) and nucleoside reverse transcriptase inhibitors (NRTIs), especially stavudine and zidovudine, are implicated as follows: [0052] (i) decreased production of retinoic acid and triglyceride uptake: PIs have a high affinity for the catalytic site of HIV-1 protease, which shares a 60% sequence homology with 2 proteins involved in lipid metabolism, cytoplasmic retinoic acid-binding protein type 1 (CRABP-1) and low-density lipoprotein receptor-related protein (LDLR-RP). Inhibition of CRABP-1 impairs the production of retinoic acid, leading to decreased fat storage and adipocyte 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 removal of chylomicrons and triglycerides from the circulation. [0053] (ii) inhibition of mitochondrial DNA (mtDNA) polymerase gamma: NRTIs inhibit mtDNA polymerase gamma, leading to mtDNA depletion, respiratory chain dysfunction, and reduced energy production, 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. [0054] (iii) inhibition of lipid metabolism: Some PIs, particularly ritonavir, inhibit cytochrome P450 3A, a key enzyme in lipid metabolism. [0055] (iv) prevention of the development of adipocytes: Saquinavir, ritonavir, and nelfinavir (all PIs) directly inhibit the development of adipocytes from stem cells and increase the metabolic destruction of fat in existing adipocytes.

[0056] In an embodiment the compound of formula (I) or pharmaceutical composition containing 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 antibiotic/probiotic or an anti-inflammatory agent. In another embodiment the pharmaceutical composition cure or prevent or alleviate at least one symptoms of lipodystrophy including, but not limited to triglyceride level, VLDL level and Apo B level in serum. In another embodiment the pharmaceutical composition cure or prevent of lipodystrophy by improving at least one of the condition selected from HDL level, Apo A1 level, HOMA of beta cell function derived from c-peptide.

[0057] In another 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 according to formula (I) can be co-administered with a therapeutic 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 control cholesterol, an antioxidant, an appetite suppressing agent, an anti-obesity 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 lipohypertrophy, lipoatrophy and Metabolic abnormalities in HIV patient.

[0058] The compound of the present invention when M+ represents K, Mg can be prepared by the processes disclosed herein below along with suitable modifications known to a skilled person.

Example 1

Preparation of (S)--Ethoxy-4-[2-[-methyl-5-[4-(methylthio)phenyl]-1H-pyrrol-1-yl]ethoxy]benzene-propanoic acid ethyl ester

[0059] In a dry, 5 L round bottom flask 2.1 L toluene was taken under nitrogen. To this 366.1 g ethyl (S)--2-ethoxy-3-(4-hydroxyphenyl)propionate was added at room temperature.

[0060] The reaction mixture was stirred under heating, using Dean-stark apparatus, to remove water azeotropically. The reaction mixture was cooled to 50 C. To this was added 319 g anhydrous potassium carbonate and stirred at 90-92 C. for 1 hr. Cooled to 65 C. and added 500 g 2-(2-methyl-5-(4-(methylthio)phenyl)-1H-pyrrol-1-yl)ethyl methanesulfonate and 22 g tetra butyl ammonium bromide. Reaction 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)--Ethoxy-4-[2-[-methyl-5-[4-(methylthio)phenyl]-1H-pyrrol-1-yl]ethoxy]benzene-propanoic acid ethyl ester.

[0061] Yield: 650 g, HPLC purity: 84.10%; % Yield 76.0%.

Example 2

Preparation of (S)--Ethoxy-4-[2-[-methyl-5-[4-(methylthio)phenyl]-1H-pyrrol-1-yl]ethoxy] benzenepropanoic acid magnesium salt

[0062] In a dry, 250 mL round bottom flask 80 mL methanol was taken. To this 20 g (S)--ethoxy-4-[2-[-methyl-5-[4-(methylthio)phenyl]-1H-pyrrol-1-yl]ethoxy]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 stirred at room temperature for 3 hours to complete 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)--ethoxy-4-[2-[-methyl-5-[4-(methylthio)phenyl]-1H-pyrrol-1-yl]ethoxy]benzenepropanoic acid magnesium salt. Solid was filtered, and dried.

[0063] Yield: 10.3 g; HPLC Purity: 98.32%; Chiral purity: 97.64%.

[0064] Following the process similar to those described in Examples 1 & 2 the following batches of )--Ethoxy-4-[2-[-methyl-5-[4-(methylthio)phenyl]-1H-pyrrol-1-yl]ethoxy] benzenepropanoic acid magnesium salt were prepared.

TABLE-US-00001 No. Batch no. Input Output % Yield HPLC purity Chiral purity 1 Example 3 10 g 5.02 g 61.21% 98.22% 98.58% 2 Example 4 10 g 4.97 g 60.68% 97.91% 3 Example 5 15 g 7.34 g 61.94% 98.20% 4 Example 6 15 g 8.38 g 67.50% 99.05% Similar reaction carried out using Magnesium chloride 5 Example 7 10 g 6.5 g 79.25% 98.53% 99.32% Similar reaction carried out using Magnesium sulfate 6 Example 8 10 g 6.8 g 82.91% 98.5%

[0065] The present invention further discloses use of said compound of formula (I) or their suitable pharmaceutical compositions for the treatment of lipohypertrophy, lipoatrophy and metabolic abnormalities in HIV patient.

Example 9

(S)--Ethoxy-4-[2-[-methyl-5-[4-(methylthio)phenyl]-1H-pyrrol-1-yl]ethoxy]benzenepropanoic acid potassium salt

[0066] In a dry, 250 mL round bottom flask 72 mL ethyl acetate was taken. To this 10 g (S)-()-1-phenylethylamine salt of (S)--ethoxy-4-[2-[-methyl-5-[4-(methylthio) phenyl]-1H-pyrrol-1-yl]ethoxy]benzene-propanoic acid was added at room temperature and subsequently 50 mL water and 4.8 mL dilute hydrochloric acid (water 1: 1:35% HCl) was added and stirred at room temperature till solid was dissolved.

[0067] Layer was separated and organic layer was washed with water, dried over sodium sulfate and solvent removed. 9.2 g oily mass obtained. To this was added 50 mL methanol and stirred under nitrogen. To this was added 1.81 g potassium t-butoxide and was stirred at room temperature for 15 min. Solvent removed and added n-Hexane. Again n-hexane was removed and added methanol. Solvent removed under vacuum. Hygroscopic material obtained. Dried it under vacuum to get (S)--ethoxy-4-[2-[-methyl-5-[4-(methylthio)phenyl]-1H-pyrrol-1-yl]ethoxy]benzenepropanoic acid potassium salt.

[0068] Yield7.6 g, (92.77%), HPLC Purity 98.60%, Chiral purity 99.56%

Example 10

[0069] 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 safety and efficacy of 4 mg of compound of formula (I) in hypertriglyceridemia in HIV associated lipodystrophy.

[0070] Methodology: This was 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.

[0071] After obtaining informed written consent, subjects with hypertriglyceridemia in HIV associated lipodystrophy, on treatment with HAART for at least 18 months and satisfying the inclusion and exclusion criteria were enrolled in the study. The subjects received 4 mg of compound of formula (I) tablet orally, once daily for 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 (I)

Dose 4 mg

[0072] Duration of treatment: 12 weeks

Mode of administration: Oral

[0073] Batch number: EMK328

Criteria for evaluation: Efficacy:

[0074] The primary efficacy endpoint was to assess the percent change in TG levels from baseline to Week 6 and Week 12. The secondary efficacy endpoint was the assessment of LDL, VLDL, HDL, Non HDL cholesterol, Total cholesterol, Apo A1, Apo B, and C-peptide and fasting insulin for HOMA beta and HOMA IR.

Safety:

[0075] Clinical examination and recording of adverse events (AEs) was done on all visits. Electrocardiogram was recorded at screening visit and at Week 12. Urine pregnancy test was conducted at screening visit

[0076] Haematological examination included haemoglobin, haematocrit, red blood cell (RBC) count, white blood cell (WBC) count with differential (neutrophils, lymphocytes, monocytes, eosinophils and basophils) and platelet count.

[0077] Biochemistry tests included AST, ALT, ALP, total bilirubin, serum proteins, total albumin and globulin, -GTT, BUN, Serum creatinine, serum uric acid, CPK, and urine R/Ms (including microalbuminuria and ketonuria).

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

Statistical Methods

[0078] For the efficacy endpoints, treatment effect was evaluated using an analysis of variance (ANOVA) model with factors for baseline and treatment. Treatment effects were estimated using the least-square means (LSM) and 95% confidence intervals (Cis) 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.

[0079] For safety analysis the frequency tabulations of abnormal physical examination and abnormal clinical laboratory parameters were presented for each visit. Summary statistics for clinical laboratory parameters and vital signs were presented for each visit.

[0080] 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 organ class (SOC) and by MedDRA preferred term for treatment emergent adverse events (TEAEs). All AEs, including those arising 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

[0081] 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 population. The results of compound of formula (I) from phase 11 studies in Dyslipidemia subjects demonstrated that compound of formula (I) 4 mg is well tolerated and effective at once daily dosing. Phase I study demonstrated food significantly affects absorption of compound of formula (I), so drug was recommended to be consumed preferably in fasting condition. Based upon these observations 4 mg once daily in fasted condition was selected for present study

Selection of Study Population

Inclusion Criteria

[0082] Subjects who satisfied all of the following criteria were eligible for enrolment in the study: [0083] 1. Males and females aged 18-65 years. [0084] 2. Confirmed diagnosis of HIV1 and on HAART for at least 18 months. [0085] 3. On stable ART regimen for at least 8 weeks prior to inclusion in the study and ART regimen not expected to change in next 3 months. [0086] 4. Subjects clinically diagnosed as HIV lipodystrophy (at least 1 moderate or severe lipodystrophy feature identified by doctor and patient, except isolated abdominal obesity) [0087] 5. Triglycerides >200 to 500 mg %. [0088] 6. CD4 count of >50/mm.sup.3 [0089] 7. Subject who had given informed consent for participation in this trial.

Treatments

Treatments Administered

[0090] The study had a single arm. Subjects received 4 mg of compound of formula (I) orally once daily in the morning before breakfast, for a period of 12 weeks.

Identity of Investigational Product(s)

[0091] Compound of formula (I) is divalent magnesium salt of carboxylic acid in the form of white, amorphous powder, which is freely soluble in dimethyl sulfoxide, dichloromethane, slightly soluble in methanol and insoluble in water. The drug was supplied as uncoated tablets of 4 mg of the active ingredient.

[0092] Supply from batch no EMK328 was used during the study. The study drug was manufactured and packaged in cGMP facility.

Primary Efficacy Variable(s)

[0093] The primary efficacy endpoint was to determine the percent change in TG levels from baseline to Week 6 and Week 12.

Secondary Efficacy Variables

[0094] The secondary efficacy endpoint was to determine the percent change in LDL, VLDL, HDL, total cholesterol, non-HDL Cholesterol (measured value), Apo A1, and Apo B, C-peptide and fasting 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

[0095] The demographic and baseline characteristics were summarized for compound of formula (I) 4 mg treatment arm. For continuous measurements such as age, the mean, median, standard deviation (SD) and range were tabulated. For categorical measurements such as gender, the frequencies were computed;

Efficacy Analyses:

[0096] The primary efficacy variable was the reduction in TG at Week 6 and Week 12 of the treatment period compared with baseline. The change from baseline was determined as the difference between the means for the treatment period (Weeks 6/Weeks 12) and the baseline.

For the efficacy endpoints, treatment effect was evaluated using an analysis of variance (ANOVA) model with factors for baseline and treatment. Treatment effects were estimated using the least-square means (LSM) and 95% confidence intervals (CIs) 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.

[0097] Intent-to-treat (ITT) and/or Per Protocol (PP) analysis were carried out for the study. The PP analysis was considered definitive while the ITT analysis was considered supportive during the trial analysis.

Efficacy Results and Tabulations of Individual Patient Data

Analysis of Efficacy

[0098] One subject identified as EHT004 in the study, a 35-year-old male, was reported with abnormally low levels of HDL (3.95 mg/L) and LDL (6.25 mg/L) at Visit 1. Though this subject completed the study and was assessable for efficacy, it was decided to exclude this subject from the efficacy analyses. Therefore a total of 49 subjects were analyzed for efficacy.

Primary Endpoints

[0099] The percent change from baseline in serum TG levels at Week 6 and Week 12 following compound of formula (I) 4 mg was statistically significant (40.984.89 and 45.113.60, respectively [p-value: <0.0001, each]) (Table 1).

TABLE-US-00002 TABLE 1 Analysis of change in Triglyceride (mg/dL) from baseline by visit COMPOUND OF Laboratory FORMULA (I) Test (Unit) Visit 4 mg (N = 49) TG Visit 1 n 49 (mg/dL) Mean SD 301.68 86.99 Median 275.45 Minimum 200.10 Maximum 481.42 Visit 3 n 49 (Week 6) Mean SD 172.81 106.30 Median 147.68 Minimum 42.61 Maximum 631.08 Change from Visit 1 128.87 14.96 (LS Mean SE) p-values <0.0001 % Change from Visit 1 40.98 4.89 (LS Mean SE) p-values <0.0001 Visit 4 n 49 (Week 12) Mean SD 166.97 89.17 Median 145.91 Minimum 46.88 Maximum 387.69 Change from Visit 1 134.71 10.78 (LS Mean SE) p-values <0.0001 % Change from Visit 1 45.11 3.60 (LS Mean SE) p-values <0.0001 Key to abbreviations: LSM = least square means; N = number of subjects in the treatment group; n = number of subjects having non-missing baseline and post-baseline values; SD = standard deviation; SE = standard error; TG = triglycerides. Note: p-values < 0.05 indicates significant and from ANOVA model

Secondary Endpoints

HDL Cholesterol:

[0100] There was an increase in the HDL cholesterol levels following administration of compound of formula (I) 4 mg. The percent change from baseline in HDL cholesterol following compound of formula (I) 4 mg at Week 6 and Week 12 was statistically significant (29.925.73 and 34.566.13, respectively [p-value: <0.0001 each]) (Table 2).

TABLE-US-00003 TABLE 2 Analysis of change in HDL Cholesterol (mg/dL) from baseline by visit COMPOUND OF Laboratory FORMULA(I) Test (Unit) Visit 4 mg (N = 49) HDL Visit 1 n 49 Cholesterol Mean SD 35.27 7.85 (mg/dL) Median 34.52 Minimum 22.23 Maximum 49.90 Visit 3 n 49 (Week 6) Mean SD 44.44 14.04 Median 43.36 Minimum 20.13 Maximum 73.50 Change from Visit 1 9.17 1.99 (LS Mean SE) p-values <0.0001 % Change from Visit 1 29.92 5.73 (LS Mean SE) p-values <0.0001 Visit 4 n 49 (Week 12) Mean SD 46.14 14.84 Median 47.70 Minimum 17.61 Maximum 82.89 Change from Visit 1 10.87 2.08 (LS Mean SE) p-values <0.0001 % Change from Visit 1 34.56 6.13 (LS Mean SE) p-values <0.0001 Key to abbreviations: LSM = least square means; N = number of subjects in the treatment group; n = number of subjects having non-missing baseline and post-baseline values; SD = standard deviation; SE = standard error; HDL = high density lipoprotein. Note: p-values < 0.05 indicates significant and from ANOVA model

C-Peptide HOMA of Insulin Resistance:

[0101] There was an increase in insulin resistance after treatment with compound of formula (I). The percent change in HOMA IR from baseline following administration of compound of formula (I) 4 mg at Week 6 and Week 12 was statistically significant (27.874.22 and 58.295.74 respectively[p-value: <0.0001 each]) (Table 3).

TABLE-US-00004 TABLE 3 Analyses of change in HOMA of insulin resistance for C-Peptide from baseline by visit COMPOUND OF Laboratory FORMULA(I) Test (Unit) Visit 4 mg (N = 49) Homa of Insulin Visit 1 n 49 Resistance Mean SD 1.59 0.82 for C-Peptide Median 1.40 Minimum 0.50 Maximum 3.80 Visit 3 n 49 (Week 6) Mean SD 1.86 0.77 Median 1.70 Minimum 0.90 Maximum 3.60 Change from Visit 1 0.27 0.05 (LS Mean SE) p-values <0.0001 % Change from Visit 1 27.87 4.22 (LS Mean SE) p-values <0.0001 Visit 4 n 49 (Week 12) Mean SD 2.15 0.62 Median 2.10 Minimum 1.10 Maximum 3.60 Change from Visit 1 0.56 0.05 (LS Mean SE) p-values <0.0001 % Change from Visit 1 58.29 5.74 (LS Mean SE) p-values <0.0001 Key to abbreviations: HOMA: homeostasis model assessment, IR: insulin resistance, LSM = least square means; N = number of subjects in the treatment group; n = number of subjects having non-missing baseline and post-baseline values; SD = standard deviation; SE = standard error Note: p-values < 0.05 indicates significant and from ANOVA model

Insulin (Fasting):

[0102] There was an increase in insulin resistance after treatment with compound of formula (I). The percent change in Insulin from baseline following administration of compound of formula (I) 4 mg at Week 6 and Week 12 was statistically significant (23.713.55 and 47.104.21 respectively [p-value: <0.0001 each]) (Table 4).

TABLE-US-00005 TABLE 4 Analyses of change in Insulin (fasting) from baseline by visit COMPOUND OF Laboratory FORMULA (I) Test (Unit) Visit 4 mg (N = 49) Insulin Visit 1 n 49 (fasting) Mean SD 9.21 6.26 u/mL Median 7.40 Minimum 2.65 Maximum 28.06 Visit 3 n 49 (Week 6) Mean SD 10.42 5.74 Median 8.35 Minimum 2.14 Maximum 26.82 Change from Visit 1 1.21 0.22 (LS Mean SE) p-values <0.0001 % Change from Visit 1 23.71 3.55 (LS Mean SE) p-values <0.0001 Visit 4 n 49 (Week 12) Mean SD 11.40 4.45 Median 10.18 Minimum 5.93 Maximum 24.29 Change from Visit 1 2.20 0.21 (LS Mean SE) p-values <0.0001 % Change from Visit 1 47.10 4.21 (LS Mean SE) p-values <0.0001 Key to abbreviations: LSM = least square means; N = number of subjects in the treatment group; n = number of subjects having non-missing baseline and post-baseline values; SD = standard deviation; SE = standard error Note: p-values < 0.05 indicates significant and from ANOVA model

Insulin HOMA of Beta-Cell Function:

[0103] There was an increase in HOMA of Beta-cell function derived from Insulin after treatment with compound of formula (I). The percent change in the HOMA of Beta-cell function derived from Insulin from baseline at Week 6 and Week 12 was statistically significant (52.5014.94 and 45.646.22, respectively [p-value: 0.0010 and <0.0001, respectively])(Table 5).

TABLE-US-00006 TABLE 5 Analyses of change in HOMA of Beta Cell Function for Insulin from baseline by visit COMPOUND OF Laboratory FORMULA (I) Test (Unit) Visit 4 mg (N = 49) HOMA of Beta Visit 1 n 48 Cell Function Mean SD 107.82 52.85 for Insulin Median 97.25 Minimum 10.20 Maximum 234.50 Visit 3 n 49 (Week 6) Mean SD 136.41 76.00 Median 116.50 Minimum 34.90 Maximum 348.00 Change from Visit 1 29.55 8.76 (LS Mean SE) p-values 0.0015 % Change from Visit 1 52.50 14.94 (LS Mean SE) p-values 0.0010 Visit 4 n 49 (Week 12) Mean SD 137.56 46.11 Median 125.60 Minimum 9.80 Maximum 273.30 Change from Visit 1 30.78 4.25 (LS Mean SE) p-values <0.0001 % Change from Visit 1 45.64 6.22 (LS Mean SE) p-values <0.0001 Key to abbreviations: HOMA: homeostasis model assessment, LSM = least square means; N = number of subjects in the treatment group; n = number of subjects having non-missing baseline and post-baseline values; SD = standard deviation; SE = standard error Note: p-values < 0.05 indicates significant and from ANOVA model

Efficacy Conclusions

Primary Endpoint:

[0104] There was a statistically significant reduction from baseline in serum TG levels at Week 6 and Week 12 following compound of formula (I) 4 mg (percent change of 40.984.89 and 45.113.60, respectively [p value: <0.0001, each])

Secondary Endpoints:

[0105] There was no statistically significant change in the non-HDL cholesterol levels from baseline following administration of compound of formula (I) 4 mg at Week 6 and Week 12 (p-values: 0.3963 and 0.4646, respectively) [0106] There was a statistically significant increase in the HDL cholesterol levels from baseline following administration of compound of formula (I) 4 mg at Week 6 and Week 12 (percent change: 29.925.73 and 34.566.13, respectively [p-value: <0.0001 each]). [0107] There was a statistically significant increase in the HOMA of Beta-cell function derived from C-peptide from baseline following administration of compound of formula (I) 4 mg at Week 6 and Week 12 (68.2525.58 and 71.6716.20, respectively [p-value: 0.0104 and <0.0001, respectively]). [0108] There was a statistically significant increase in the HOMA of insulin resistance derived from insulin from baseline after treatment with compound of formula (I) at Week 6 and Week 12 (percent change: 29.103.94 and 42.653.79, respectively [p-value: <0.0001 each]).

[0109] Therefore, the compound of the present invention including pharmaceutical compositions containing the same was found to be useful for the treatment of lipohypertrophy, lipoatrophy and Metabolic abnormalities in HIV patients.