OA17459A - A novel composition for nonalcoholic fatty liver disease (NAFLD). - Google Patents

Abstract

The present invention provides a compound of formula (I) or pharmaceutical acceptable thereof, wherein 'R' is herein described. In addition, the invention relates to composition comprising effective therapeutic amount of compound of formula (I) and methods of using the compounds for treating or prevention disorder such as nonalcoholic fatty liver disease (NAFLD) including fatty liver (steatosis), nonalcoholic steatohepatitis (NASH), and cirrhosis (advanced scarring of the liver).

Description

The présent invention relates to pharmaceutical compositions containing the formula (I)

for the réduction and removal of lipid accumulated in the liver cells (hépatocytes) which is associated with nonalcoholic fatty liver disease (NAFLD). The présent invention further provides the composition of formula (I) useful in the prévention and treatment of nonalcoholic fatty liver disease (NAFLD)

Backgronnd of the Invention

Nonalcoholic fatty liver disease (NAFLD) refers to a wide spectrum of liver disease ranging from simple fatty liver (steatosis), to nonalcoholic steatohepatîtis (NASH), to cirrhosis (irréversible, advanced scarring of the liver). Ail of the stages of NAFLD hâve in common the accumulation of fat (fatty infiltration) in the liver cells (hépatocytes). In NASH, the fat accumulation is associated with varying degrees of inflammation, fibrosis and scarring of the liver.

NASH and NAFLD are frequently reported in both men and women, although it most often appears in women and is especially prévalent in the obese. Although the disease has been observed to be accompanied by several other pathological conditions, including diabètes mellitus, hyperlipidemia, hyperglycemia, ail part of the metabolic syndrome, the cause and progression of the disease, as well as the causal or temporal relation to these conditions, is not well understood.

However, in patients suffering from NAFLD and NASH in particular, certain characteristics of liver tissue and abnormalities of function are typical.

Specifically, fatty deposits, tissue degeneration, inflammation, cell degeneration, fibrosis, -_Az l

cirrhosis, élévation of free fatty acids and other such abnormalîties hâve corne to be associated with nonalcoholîc steatohepatîtis and are frequently seen in patients suffering from different forms of NAFLD.

The physiological condition that most commonly accompanies NASH is obesity, with approximately 70% and above of NASH sufferers also displaying clinically diagnosed obesity. NASH is particularly prévalent in obese patients who hâve undergone jejunal bypass to treat the obesity. In NASH patients, the extent of obesity tends to be generally correlated with the amount of steatosis and to be unrelated to non-insulindependent diabètes mellitus. However, non-insulin-dependent diabètes mellitus increases 10 the prevalence of steatohepatîtis especially în patients requiring insulin. Unless a massive amount of the excess body weight is eliminated, weight loss in patients before death does not appear to alleviate the steatosis and, somewhat paradoxically, obese patients who lost weight before death can hâve a higher incidence of steatohepatîtis.

Even in NASH patients who do not consume any alcohol at ail, liver biopsy 15 specimens tend to mimic those seen în patients suffering from alcoholic hepatitis. However, a comparison of the two conditions reveals a higher incidence of vacuolation (indicative of diabètes) and steatosis în NASH as compared to alcoholic hepatitis. Patients suffering from alcoholic hepatitis also hâve a higher incidence of periportal and pericellular fibrosis and prolifération of the bile ducts. Overall, the symptoms and 20 histological damage observed in alcoholic hepatitis patients are more severe than in NASH.

Currently, there is no established therapy for patients suffering from NASH. Weight loss is a common prescription, simply because obesity is frequently detected in patients suffering from NASH. The effect of a réduction in weight loss on NASH cannot 25 be determined with certainty, however, because obese patients seldom maintain significant weight réduction. Thus, there is a need to find a treatment for NAFLD and particularly NASH.

Objects of the Invention

In one embodiment, the présent invention discloses a pharmaceutical composition containing the compound of the Formula (I)

for réduction and removal of lipid accumulated in the liver cells (hépatocytes), required for treating and preventing certain diseases and conditions related to nonalcoholic fatty liver disease (NAFLD) încluding fatty liver (steatosis), nonalcoholic steatohepatitis (NASH), and cîrrhosis (advanced scarring of the liver) in patient in need of such treatment.

In another embodiment the présent invention provides a method and a formulation comprising an effective amount of compound of Formula (I) for treating nonalcoholic fatty liver disease (NAFLD) încluding fatty liver (steatosis), nonalcoholic steatohepatitis (NASH), and cîrrhosis (advanced scarring of the liver).

The method comprises administering to a subject an effective amount of a compound of formula (I) as a pharmaceutical formulation, as disclosed hereinafter încluding pharmaceutically acceptable salts of the compound of formula (I).

In yet another embodiments the invention further provides a pharmaceutical composition containîng effective amount of compound of formula (I) suitable for treatment of nonalcoholic fatty liver disease (NAFLD) încluding fatty liver (steatosis), nonalcoholic steatohepatitis (NASH), and cîrrhosis (advanced scarring of the liver).

In another embodiment the présent invention provides a method of treating alcoholîc steatohepatitis in a subject, comprising administering to the subject an effective amount of a compound according to Formula (I), or a pharmaceutically acceptable sait thereof as a suitable pharmaceutically acceptable composition.

In another embodiment the présent invention provides a method of treating liver failure in a subject, comprising administering to the subject an effective amount of a compound according to Formula (I), or a pharmaceutically acceptable sait thereof.

The above and other embodiments of the présent invention are disclosed further hereinafter.

Description of the Figures:

Figure l: Effect of magnésium sait of compound of formula (I) wherein R is -SMe on ALT in PP population

Figure 2: Effect of magnésium sait of compound of formula (I) wherein R is -SMe on liver function test in Safety Population

Figure 3: Effect of magnésium sait of compound of formula (I) wherein R is -SMe on C-Peptide and HOMA Function in PP population

Figure 4: Effect of magnésium sait of compound of formula (I) wherein R is -SMe on Triglycérides in PP population

Detaïled Description of the Invention

The présent invention describes a pharmaceutical composition for réduction and removal of lipid accumulated in the liver cells (hépatocytes), required for treating and preventing certain diseases and conditions in subject suffering from nonalcoholic fatty liver disease (NAFLD) including fatty liver (steatosis), nonalcoholic steatohepatitis (NASH), and cirrhosis (advanced scarring of the liver) and methods for améliorai ing and/or treating such disease conditions.

The formulation comprises compound of formula (I) and the method comprises administering to a subject in need thereof an effective amount of a compound according to Formula (I), or a pharmaceutically acceptable sait thereof.

(I) wherein ‘R’ is selected from hydroxy, hydroxyalkyl, acyl, alkoxy, alkylthio, thioalkyl, aryloxy, arylthîo and M⁺ represents suitable métal cations such as Na\ K⁺, Ca⁺², Mg'² and the like.

Définitions and Abbreviations

As used above, and throughout this disclosure, the following terms, unless otherwise indîcated, shall be understood to hâve the following

Meanings :

Patient includes both human and animais. Mammal means humans and other mammalian animais.

A subject is a mammal, preferably a human, but can also be an animal in need of veterinary treatment, e.g., companîon animais (e.g., dogs, cats, and the like), farm animais (e.g., cows, sheep, pigs, horses, and the like) and laboratory animais (e.g., rats, mice, guinea pigs, and the like).

As used herein treating includes achieving, partially or substantially, one or more of the following results: partially or totally reducing the extent of the disease, disorder or syndrome (e.g., reducing fat deposits, increasing însulin activity/sensitivîty, reducing weight); ameliorating or împroving a clinical symptom or indicator associated with the disorder; delaying, inhibiting or preventing the progression of the disease, disorder or syndrome; or partially or totally delaying, inhibiting or preventing the onset or development of disorder. Delaying, inhibiting or preventing the progression of the disease, disorder or syndrome includes for example, delaying, inhibiting or preventing the progression of fatty liver to NASH; delaying, inhibiting or preventing the progression of NASH to cirrhosis, end-stage liver disease and/or hepatocellular carcinoma; and delaying, inhibiting or preventing the progression of pre-dîabetes to diabètes.

The term “alkyl” used herein, either alone or in combination with other radicals, dénotés a linear or branched radical containing one to twelve carbons, such as methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, tert-butyl, amyl, t-amyl, n-pentyl , n-hexyl, iso-hexyl, heptyl, octyl and the like.

The term “alkoxy” used herein, either alone or in combination with other radicals, dénotés a radical alkyl, as defined above, attached directly to an oxygen atom, such as methoxy, ethoxy, n-propoxy, iso-propoxy, n-butoxy, t-butoxy, iso-butoxy, pentyloxy, hexyloxy, and the like.

The term “aryl” or “aromatic” used herein, either alone or in combination with other radicals, refers to an optionally substituted aromatic system containing one, two or three rings wherein such rings may be attached together in a pendant manner or may be fused, such as phenyl, naphthyl, tetrahydronaphthyl, indane, biphenyl, and the like. The term ‘aralkyl” dénotés an alkyl group, as defined above, attached to an aryl, such as benzyl, phenethyl, naphthylmethyl, and the like. The term “aryloxy” dénotés an aryl radical, as defined above, attached to an alkoxy group, such as phenoxy, naphthyloxy and the like, which may be substituted. The term “aralkoxy” dénotés an arylalkyl moiety, as defined above, such as benzyloxy, phenethyloxy, naphthylmethyloxy, phenylpropyloxy, and the like, which may be substituted.

The term “acyl” used herein, either alone or în combination with other radicals, refers to a radical containing one to eight carbons such as formyl, acetyl, propanoyl, butanoyl, iso-butanoyl, pentanoyl, hexanoyl, heptanoyl, benzoyl and the like, which may be substituted.

The term “hydroxyalkyl” used herein, either alone or in combination with other radicals, refers to an alkyl group, as defined above, substituted with one or more hydroxy radicals, such as hydroxymethyl, hydroxyethyl, hydroxypropyl, hydroxybutyl, hydroxypentyl, hydroxyhexyl and the like.

The term “thio(Ci-Ci2)alkyl” or “thio((C|-C6)alkyl” used herein, either alone or în combination with other radicals, represents an alkyl group, as defined above, attached to a group of formula -SR’, where R’ represents hydrogen, alkyl or aryl group, e.g. thiomethyl, methylthiomethyl, phenylthiomethyl and the like, which may be substituted

Effective amount or therapeutically effective amount is meant to describe an amount of compound or a composition of the présent invention effective in inhibitîng the abovenoted diseases and thus producing the desired therapeutic, ameliorative, inhibitory or preventative effect.

One or more compounds of the invention may exist in unsolvated as well as solvated forms with pharmaceutically acceptable solvents such as water, éthanol, and the like, and it is intended that the invention embrace both solvated and unsolvated forms. Solvaté means a physical association of a compound of this invention with one or more solvent molécules. This physical association involves varying degrees of ionic and covalent bonding, including hydrogen bonding. In certain instances the solvaté will be capable of isolation, for example when one or more solvent molécules are incorporated în the crystal lattice of the crystalline solid. Solvaté encompasses both solution- phase and isolatable solvatés.

One or more compounds of the invention may optionally be converted to a solvaté. Préparation of solvatés is generally known. Thus, for example, M. Caira et al, J. Pharmaceutical Sci, 93(3), 601-611 (2004) describe the préparation of the solvatés of the antifungal fluconazole in ethyl acetate as well as from water. Similar préparations of solvatés, hemisolvate, hydrates and the like are described by E. C. van Tonder et al, AAPS PharmSciTech., 5(l), article 12 (2004); and A. L. Bingham et al, Chem. Commun., 603604 (2001)

The compounds of Formula (I) can form salts which are also within the scope of this invention. Reference to a compound of Formula (I) herein is understood to include reference to salts thereof, unless otherwise indicated. The term salt(s), as employed herein, dénotés acidic salts formed with inorganic and/or organic acids, as well as basic salts formed with inorganic and/or organic bases. In addition, when a compound of Formula (I) contain both a basic moiety, such as, but not limited to a pyridine or imidazole, and an acidic moiety, such as, but not limited to a carboxylic acid, zwitterions (inner salts) may be formed and are also included within the term salt(s) as used herein.

Pharmaceutically acceptable (i.e., non-toxic, physiologically acceptable) salts are preferred, although other salts are also useful. Salts of the compounds of the Formula (I) may be formed, for example, by reacting a compound of Formula I with an amount of acid or base, such as an équivalent amount, in a medium such as one in which the sait précipitâtes or in an aqueous medium followed by lyophilization.

Exemplary acid addition salts include acétates, ascorbates, benzoates, benzenesulfonates, bisulfates, borates, butyrates, citrates, camphorates, camphorsulfonates, fumarates, hydrochlorides, hydrobromîdes, hydroîodîdes, lactates, maleates, methanesulfonates, naphthalenesulfonates, nitrates, oxalates, phosphates, propîonates, salicylates, succinates, sulfates, tartarates, thiocyanates, toluenesulfonat.es (also known as tosylates,) and the like.

Exemplary basic salts include ammonium salts, alkali métal salts such as sodium, lithium, and potassium salts, alkaline earth métal salts such as calcium and magnésium salts, salts with organic bases (for example, organic amines) such as dicyclohexylamines, t-butyl amines, and salts with amino acids such as arginine, lysine and the like. Basic nitrogen-containing groups may be quartemized with agents such as lower alkyl halides (e.g., methyl, ethyl, and butyl chlorides, bromides and iodides), dialkyl sulfates (e.g., dimethyl, diethyl, and dibutyl sulfates), long chain halides (e.g., decyl, lauryl, and stearyl chlorides, bromides and iodides), aralkyl halides (e.g., benzyl and phenethyl bromides), and others.

Ail such acid salts and base salts are intended to be pharmaceutically acceptable salts within the scope of the invention and ail acid and base salts are considered équivalent to the free forms of the corresponding compounds for purposes of the invention

Polymorphie forms of the compounds of Formula (I), and of the salts, solvatés, esters and prodrugs of the compounds of Formula (I) are intended to be included in the présent invention.

As used herein, the term composition is intended to encompass a product comprising the specified ingrédients in the specifîed amounts, as well as any product which results, directly or îndirectly, from combination of the specified ingrédients in the specified amounts

In the embodiments the présent invention provides a suitable pharmaceutical composition of compounds of formula (I) or their dérivatives, which comprises one or more pharmaceutical excipients, antioxidants and chelating agents, wherein the pH of the composition is above 6, preferably in the range from about pH 6 to pH of about 10.

In such embodiments the pharmaceutical composition of the présent invention essentially comprises of • the pharmaceutically active substance;

• Suitable additives;

• a suitable stabilizer;

• optionally with one or more pharmaceutically acceptable excipients. One function of the liver is to process fats and proteins from digested food.

Fatty liver disease covers a range of conditions where there is a build-up of fat in the liver cells. The liver cells (hépatocytes) normally contain some fat and related fatty chemicals (triglycérides, fatty acids, etc). Excess fat is normally passed out of liver cells, into the bloodstream, and then taken up and stored in fat cells (adipose cells) throughout the body. In fatty liver disease, excess fat builds up in liver cells. This is thought to happen if there is some problem or disruption in the normal processing of fat and related fatty chemicals in the liver cells. Simple fatty liver (also called hepatic steatosis) is présent when the fat content inside liver cells makes up more than 5-10% of the liver's weight. Simple fatty liver is not associated with serious damage or harm to the liver.

Nonalcoholic fatty liver disease (NAFLD) refers to a wide spectrum of liver disease ranging from: i) simple fatty liver (steatosis), in which there are fat deposits on the liver; iî) nonalcoholic steatohepatitis (NASH) in which there are fat deposits on the liver along with inflammation and damage of the liver; and îii) cîrrhosis in which there is irréversible, advanced scarring of the liver.

Ail of the stages of NAFLD hâve in common the accumulation of fat (fatty infiltration) in the liver cells (hépatocytes). Fatty liver (steatosis) can progress to nonalcoholic steatohepatitis (NASH). In NASH, the fat accumulation is associated with varying degrees of inflammation and scarring of the liver, and in many cases insulin résistance, dyslipidemia and hypertension. NASH can progresses to fibrosing, steatohepatîtis and may trigger cirrhosis, end-stage liver disease, acute live failure and hepatocellular carcinoma. It most often occurs in people with excess body weight, elevated blood lipîds, such as cholestérol and triglycérides, and insulin résistance.

The présent invention also provides methods of treating liver failure. Acute liver failure occurs when the cells in the liver die or become damaged in a short period of time. This causes the liver to fail to work normally and can be fatal.

Any progressive liver disease, such as cirrhosis, can resuit in liver failure. Signs of liver failure include encephalopathy (altered brain function, jaundîce, ascites, fetor hepaticus and failure of coagulation).

Many people with simple fatty liver hâve other conditions where fatty liver is a complication. Many cases of simple fatty liver develop in people who drink more alcohol than the recommended Iimits. Over half of people who drink heavily develop simple fatty liver. In these cases simple fatty liver can progress to alcoholic steatohepatîtis. In this condition the excess fat in the liver cells is associated with, or may cause, inflammation of the liver. Alcoholic steatohepatîtis, may eventually cause scarring (cirrhosis) of the liver.

Effective amounts of such compounds are administered to a subject with one or more of these conditions.

In an embodiments the compounds according to Formula (I) can be used alone or în combination e.g., as an adjunct therapy, with at least one other therapeutic agent. Compounds according to formula (I) can be subject with NASH, a compound according to formula (I) can be co-administered with a therapeutic agent used to reduce one or more of the symptoms of NASH or NAFLD 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 or control cholestérol, an antioxidant, an appetite suppressîng agent, an anti-obesity agent, to control blood glucose levels, such as, sulfonylureas, an antibiotic/ probiotic or an antiinflammatory agent. Examples of such agents are listed herein and includes chlorpropamide, glipizîde, glyburide, and glimepiride; meglitinîdes, such as, repaglinide and nateglinide; biguanides, such as, metformîn and acarbose; thiazolîdinediones, such as, rosiglitazone, and pioglitazone; and insulin and its dérivatives, such as, pramlintide, exenatide, humalog, novolog, humulin, novolin, ultralente, and lanrus; an agent used to ιχτ~ control lipid levels, such as, vytorin, Clofibrate and Gemfibrozil, a plasma HDL-raisîng agent, a cholestérol lowering agent, a cholestérol biosynthesis inhibitor, for example an HMG-CoA reductase inhibitor (such as a statin, such as, Atorvastatin, Fluvastatin, Lovastatin, Pravastatin, Rosuvastatin, Simvastatin); an HMG-CoA synthase inhibitor, an acyl-coenzyme A: cholestérol acyltransferase (ACAT) inhibitor, such as, melinamide; probucol, niacin (nicotinic acid, Vitamin-B-3), nicotinic acid and the salts thereof and niacinamide; a cholestérol absorption inhibitor such as ezetimibe; a bile acid séquestrant, such as, cholestyramine, colestipol, and Colesevelam; fibrates such as clofibrate, fenofibrate, and gemfibrizol, vitamin B6 (also known as pyridoxine) and physiologically acceptable salts thereof, such as the HCl sait; vitamin B12 (also known as cyanocobalamîn), and angiotensin II antagonist convertîng enzyme inhibitor; a betablocker; an agent used to reduce weight or suppress appetite, such as, sibutramine, orlistat and the like.

In an embodiment, when methods of the présent invention is used to treat a subject with alcoholîc steatohepatitis, a compound according to formula (I) can be coadministered with a therapeutic agent used to reduce one or more of the symptoms of alcoholic steatohepatitis 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 cholestérol, an antioxidant, an appetite suppressing agent, an anti-obesity agent, an antibiotic or an anti-inflammatory agent, such as those described above.

In another embodiments when used in the methods of the présent invention to treat a subject with liver failure, a compound according to formula (I) can be co-administered with a therapeutic agent used to reduce one or more of the symptoms of alcoholic steatohepatitis including, but not limited to, an agent used to control blood glucose levels, an agent used to control lipid levels, such as those described above.

In a further embodiment, the présent invention discloses a suitable pharmaceutical composition of the compound of formula (I), for the treatment of one or more of the diseases disclosed above. A preferred pharmaceutical composition of the compound of formula (I) comprises of • the pharmaceutically active substance;

• Suitable additives;

• a suitable stabilîzer;

• optionally with one or more pharmaceutically acceptable excipients.

Each of the components may be selected from those known in the art.

In an embodiment suitable stabilizers may be selected from the classes of antioxidants or chelating agents.

In an embodiment the pharmaceutical excepients according to the présent invention can be selected from solubilizers, diluents, fillers, disintegrants, binder, lubricants, glidants, wetting agents, solvents and the like as is known in the art.

In an embodiment suitable additives are selected from sodium benzoate, sodium hydroxide, sodium sulfite and sodium carbonate.

In an embodiment antioxidants used according to the présent invention include, but are not limited to citric acid, alpha tocopherol, sodium sulphite, sodium metabisulphite, butylated hydroxy anisole (BHA), BHT (2,6-di-tert-butyl-4-m ethyl phénol), monothioglycerol, Vitamin C (ascorbic acid), and propyl gallate and combinations thereof and other similar material known to those of ordinary skilled in the art.

Chelating agent used according to the présent invention include, but are not limited to Disodium EDTA, citric acid and or its salts, maleic acid, chlorambutol, chlorhexidine or its salts, chlorocresol, combinations thereof and other similar material known to those of ordinary skill in the art.

As used herein, the term binders îs intended to mean substances used to cause adhesion of powder particles in tablet granulations. Such compounds include, by way of example and without limitation, acacia alginic acid, tragacanth, carboxymethylcellulose sodium, poly (vinylpyrrolidone), compressible sugar (e.g., NuTab), ethylcellulose, gelatin, liquid glucose, methyl cellulose, povidone and pregelatinized starch, combinations thereof and other similar material known to those of ordinary skill in the art.

When needed, other binders may also be included in the présent invention. Exemplary binders include starch, poly(ethylene glycol), guar gum, polysaccharide, bentonites, sugars, invert sugars, poloxamers (PLURONIC F68, PLURONIC FI27), collagen, albumin, celluloses in non-aqueous solvents, and the like or their suitable combinations. Other binders which may be included may be, for example, poly(propylene glycol), polyoxyethylene-polypropylene copolymer, polyethylene ester, polyethylene sorbitan ester, polyfethylene oxide), microcrystalline cellulose, poly(vinylpyrrolidone), combinations thereof and other such materials known to those of ordinary skill in the art. As used herein, the term diluent or filler is intended to mean inert substances used as ll fillers to create the desired bulk, flow properties, and compression characteristics in the préparation of tablets and capsules. Such compounds include, by way of example and without limitation, dibasic calcium phosphate, kaolin, sucrose, mannitol, microcrystalline cellulose, powdered cellulose, precîpitated calcium carbonate, sorbitol, starch, combinations thereof and other such materials known to those of ordinary skill in the art.

As used herein, the term glidant is intended to mean agents used in tablet and capsule formulations to improve flow-properties during tablet compression and to produce an anti-caking effect. Such compounds include, by way of example and without limitation, colloïdal silica, calcium silicate, magnésium silicate, silicon hydrogel, comstarch, talc, combinations thereof and other such materials known to those of ordinary skill in the art.

In an embodiment, the tenu lubricant is intended to mean substances used in tablet formulations to reduce friction during tablet compression. Such compounds include, by way of example and without limitation, calcium stéarate, magnésium stéarate, minerai oil, stearic acid, zinc stéarate, suitable combinations thereof and other such materials known to those of ordinary skill in the art.

In an embodiment, the term disîntegrant îs intended to mean a compound used in solid dosage forms to promote the disruption of the solid mass into smaller particles whîch are more readily dîspersed or dissolved. Exemplary disintegrants include, by way of example and without limitation, starches such as corn starch, potato starch, pre-gelatînized and modified starched thereof, sweeteners, clays, such as bentonite, microcrystalline mi Λ ¹ i¹3L cellulose (e.g. Avicel .), carsîum (e.g. Amberlite. .), alginates, sodium starch glycolate, gums such as agar, guar, locust beau, karaya, pectin, tragacanth, combinations thereof and other such materials known to those of ordinary skill in the art.

In an embodiment, the term wetting agent is intended to mean a compound used to aid in attaining intimate contact between solid particles and liquids. Exemplary wetting agents include, by way of example and without limitation, poloxamers, gelatin, casein, Glycerol mono-oleate, lecithin (phosphatides), gum acacia, cholestérol, tragacanth, stearic acid, benzalkonium chloride, calcium stéarate, glycerol monostearate, cetostearyl alcohol,sodium lauryl sulphate, sodium dodecyl sulfate, salts of bile acids (taurocholate, glycocholate, cholate, deoxycholate, etc.), cetomacrogol emulsifying wax, sorbitan esters, polyoxyethylene alkyl ethers (e.g., macrogol ethers such as cetomacrogol 1000), polyoxyethylene castor oil dérivatives, polyoxyethylene sorbitan fatty acid esters, (e.g., TWEEN), polyethylene glycols, polyoxyethylene stéarates colloïdal silicon dioxide, phosphates, sodium dodecylsulfate, carboxymethylcellulose calcium, carboxy methyl cellulosesodium,methyl cellulose,hydroxyethylcellulose, hydroxylpropyl cellulose, hydroxy propyl methyl cellulose phthalate, noncrystalline cellulose, magnésium aluminum silicate, triethanolamine, polyvinyl alcohol, and poly vinyl pyrrolidone (PVP) and their suitable combinations and other such materials known to those of ordinary skill in the art. Tyloxapol (a nonionic liquid polymer of the alkyl aryl polyether alcohol type, also known as superinone or triton) is another useful wetting agent which may be used.The stable pharmaceutical composition according to the présent invention may be in the form of tablet or capsule or a powder or a suspension in a liquid or an aérosol formulation or solutions, preferably in the form of tablet or capsule.

In another embodiment of the présent invention, is a described process for the préparation of a stable pharmaceutical composition of compounds of formula (I) or their dérivatives.

The stable pharmaceutical composition may be made by direct compression, wet granulation or dry granulation methods by techniques known to persons skilled in the art. Thus, for example,

In wet granulation process, the drug is mixed with one or more pharmaceutical excepîents and granulated with suitable binding solution as described earlîer, to form wet granules, the wet granules are dried and optionally sieved. The dried granules are mixed with one or more suitable excipients from those described elsewhere and then compressed into tablets or filled into capsules.

In direct compression process, the drug îs mixed with ail the pharmaceutical excipients required and then is either compressed into tablets or filled in capsules.

In dry granulation process the drug is mixed with one or more pharmaceutical excipients and compressed into slugs and these slugs are passed through required sieve. The sieved granules are mixed with one or more suitable excipients from those described elsewhere and then compressed into tablets or filled into capsules.

One or more solvents used in the formulation are selected from acetone, chloroform, dichloromethane, ethyl alcohol, ethyl acetate, methyl alcohol, isopropyl alcohol and combinations thereof and other such materials known to those of ordinary skill in the art.

In an embodiment, the compound of formula (I) or pharmaceutical compositions containing the compound of formula (I) is given to a subject in need thereof at a dose of

I3 about 0.5 mg to 5 g. A skilled person is aware how to décidé the optimum dose based on the patient profile, the severity of disease, the presence of secondary medicines and the like.

The compound of formula (I), when R is -SMe and M+ is Mg, is dosed to patients in need thereof for the treatment of one or more of the diseases described above as per the following general protocol:

Study design and Protocol:

Title of the Study - “A prospective, multi-centric, open-label, single arm study to evaluate the safety and efficacy of 4 mg of magnésium sait of compound of formula (I) wherein R is -SMe in a pharmaceutical composition as described above in Non-alcoholîc steatohepatitîs.

Objectives:

To evaluate the safety and efficacy of 4mg of magnésium sait of compound of formula (I) wherein R is SMe în Non-alcoholîc steatohepatitîs (NASH).

The following effficacy parameters were measured:

Primary Efficacy (Time frame 6 and 12 weeks):

l. Change in alanine amînotransferase (ALT) from baseline

Secondary Efficacy (Time frame 6 and 12 weeks):

• Sustained réduction in ALT level.

• C-peptide test for homeostasis model assessment (HOMA) beta and HOMA IR • Triglycéride (TG)

Criteria for Safety:

1. General and Systemic Clinîcal Examination: Cardiovascular system (CVS), respiratory system (RS), gastro-intestinal System (GIS), central nervous system (CNS) etc.

2. Laboratory Investigations: Complété blood count (CBC), aspartate aminotransferase(AST), ALT, alkalîne phosphatase (ALP), sérum bilirubîn, γglutamyl transpeptidase (GGT), Sérum proteins, blood urea nitrogen (BUN), Sérum créatinine, créatinine phosphokinase (CPR), fasting plasma glucose (FPG).

3. Frequency and severity of adverse events (AEs) for ail subjects enrolled were recorded. AU AEs, were classified using • causality • severity L· • seriousness

Methodology:

It is an interventions l, single arm, safety and effîcacy study to explore effect of magnésium sait of compound of formula (I) wherein R is -SMe suitably formulated as described above, on NASH.

Subjects was diagnosed by biopsy as suffering from NASH in last one year and willing to participate in study were invited for screening programme for inclusion in the study.

Subjects satisfying inclusion exclusion criteria will be enrolled in the study.

AU subjects were given suitable formulation of magnésium sait of compound of formula (I) wherein R is -SMe 4 mg for 12 weeks. Lifestyle modification was continued as before the study. Patient was monitored for safety and effîcacy of magnésium sait of compound of formula (I) wherein R is -SMe.

Study Schedules:

Informed consent was obtained before any trial related activity.

• Visit 1, Screening Visit/ Enrolment [Week -1 to 0]

Subjects were screened for the inclusion and exclusion criteria and those qualifying were invited to participate in the study. Clinîcal évaluation was done for baseline characteristics and anthropometry

After Clinîcal évaluations, ail baseline safety and effîcacy parameters were recorded as per Table (I) given below. Ail laboratory investigations were carried out after ovemight fasting.

During the 12 week program, a desîgnated person from the centre could interview the subjects for hîs/her general health, telephonically.

Enrolled Subjects would receive the study médication for next two weeks.

Patients were advîsed to follow same lifestyle modifications during study period as before the study.

• Visit 2, [Week 2]

Subjects were clinically examined and given the study médications for four weeks and also safety parameters were assessed as per Table (I)given below.

• Visit 3 [Week 61

Subjects were clinically examined and given the study médications for next 6 weeks.

Safety and effîcacy parameters were assessed as per Table (I) given below.

• Visit 4 IWeek 121

Subjects were clinically examined and safety and efficacy parameters were assessed as per Table (I) given below.

If further investigations are required in case of any AE, investigator will be advised to assess the AE and take necessary action, if required. Subjects will be advised to contact the investigator for any complaints within next two weeks.

During the above period, if any subject misses the drug administration up to 3 consecutive days, it will not be considered drop-out or protocol déviation.

Visit and Investigation Schedule

Inclusion / Exclusion criteria '.· 'Medical History

Cliniçal ExaminatjonjA Laboratory studies (efficacy^

--¹·· -«?&·

Lipid profile, C peptide, ALT) '-ii?

Laboratory studj

ECG ,·

s...

CSG

Pregnancy _ test for female ï ...^e. -._vf subjects >.(advise ^v ^for contracep

Dispensing Médication ai. .______., , _____„___

Study ⁵ Médication capsule

Count
RecordingôfAdverse Events
Global AssessmentsY^^^I^Y'4
Study Completion
* Laboratory Tests: • Biochemistry (laboratory) parameters to be performed include following tests: • Liver Function test (LFT): AST, ALT, ALP, total bilirubin, sérum proteins, total albumin and globulin, GGT • Rénal function test: Blood urea nitrogen (BUN), Sérum créatinine and calculated GFR • Créatinine phosphokînase (CP K) • CBC

CRITERIA FOR INCLUSION/EXCLUSION:

INCLUSION CRITERIA • Subject has given informed consent for participation in this trial · Biopsy proven NASH (Biopsy done in last one year).

• ALT > than l .5 tîmes upper normal limit.

• Patient presently on lifestyle modification for NASH for at least one month.

• BMI between 23 to 40 kg/m³ • Compensated liver disease with the following hématologie, biochemical, and serological criteria on entry into protocol:

• Hemoglobin > 9 gm/dL • White blood cell (WBC) > 2.5 K/UL • Neutrophil count > 1.5 K/UL • Platelets > 100 K/UL • Sérum bilirubin, < l .5mg% • Albumin > 3.2 g/dL • Sérum créatinine within normal limits

EXCLUSION CRITERIA:

• Pregnancy and lactation • Subjects with history of gall stone • Subjects with history of myopathies or evîdence of active muscle diseases • Subject with history of alcohol consumption > than 20 gm/week and/or drug abuse • Known allergy, sensitivity or intolérance to the study drugs and their formulation ingrédients.

• Participation in any other clinical trial in past 3 months • History of malignancy; active neoplasm.

• Previous liver biopsy that demonstrated presence of cirrhosis or radiologie imaging consistent with cirrhosis or portai hypertension • Type 2 diabètes treated with agents other than the secretagogues (these include, insulin thiazolidinediones, alpha-glucosidase inhibitors, exenatide, pramlintide). Metformin will be allowed provided dose is stable science last 6 months.

• Evidence of poorly-controlled diabètes [glycosylated hemoglobin (HbAlc) > 9%].

• Type I diabètes mellitus.

• Abnormal PT/INR (Prothrombin Time/Intemational normalized ratio) • Patient on fibrates (Other antidyslipidemic drugs will be allowed provided dose is stable in last 6 months) • Use of drugs associated with a clinical or histological pîcture consistent with fatty liver disease or NASH for more than 12 consecutive weeks in the I year prior to start of the study; (these include amiodarone, tamoxifen, methotrexate, glucocorticoîds, anabolic steroids, tetracyclines, estrogens, val proate/val proie acid, chloroquine, antîHIV drugs etc.) • History of thyroid disease poorly controlled on prescribed médications • History of, or current cardiac dysrhythmias and / or a history of cardiovascular *

disease, including myocardial infarction, except patients with only well controlled hypertension.

• History of bariatric surgery, or undergoing évaluation for bariatric surgery.

• History or other evidence of severe illness or any other conditions that would make the patient, in the opinion of the investigator, unsuitable for the study (such as poorly controlled psychiatrie disease, coronary artery disease, or active gastrointestinal conditions that might interfère with drug absorption) — «

• Subject on any treatment with other drugs claimed for treatment of NASH (Pentoxyphyllin, Ursodeoxycholic acid, acetyl cholinestérase enzyme (ACE) inhibitors antioxidants such as vitamin E, vitamin C, glutathione, alpha-tocopherol, or non-prescribed complementary alternative médications (including dietary suppléments, megadose vitamins, herbal préparations, and spécial teas).) or any medicine in clinical trials for NASH, • Other cause of chronic liver disease [autoimmune, primary biliary cirrhosis, hepatitis B virus (HBV), Wilson, alpha-1-antitrypsin déficit, hemochromatosis etc.] i.e. Antinuclear antibodies (ANA) > l :l 60, Anti-smooth muscle Ab positive >1:160 , Sérum hepatitis B surface antigen (HepBsAg) positive, Sérum hepatitis C antibody (HepC Ab) positive, transferrm saturation > 45%

Results

Subjects were screened for inclusion in the study after obtaining informed consent, out of which 32 subjects were enrolled into the study. Out of these 32 subjects, 29 subjects completed the study.

The effect of magnésium sait of compound of formula (1) wherein R is -SMe at week 12 on various parameters of liver function is as below.

• There was statistically significant réduction in the ALT levels from baseline in magnésium sait of compound of formula (I) wherein R is -SMe 4 mg treatment group in the PP population at visit 3 and visit 4. (Figure 1) • There was sustained réduction in ALT levels 63.16% and 78.95% of patients at visit 3 and visit 4 respectively as per PP.

• Magnésium sait of compound of formula (I) wherein R is -SMe showed a statistical significant decrease in the Aspartate Transaminase, gamma-glutamyl transpeptidase and alkaline phosphatase at Week 3 and at Week 4 in the Safety Population. (Figure 2) • There was a non-signîficant change in the C peptide levels in magnésium sait of compound of formula (I) wherein R is -SMe 4 mg at 6 and 12 weeks as per PP analysis and non-statistically significant réduction in HOMA -Beta cell function, HOMA - Insulin Résistance. (Figure 3) • Baseline TG < 150 mg/dl - There was decrease in triglycéride in magnésium sait of compound of formula (I) wherein R is -SMe 4 mg at week 12 as compared to baseline but it was not statistically significant in the PP population.(Figure 4) • Baseline TG >150 mg/dl - While decrease was observed in sérum triglycérides at week 6 and 12 as compared to baseline but it was not statistically significant in magnésium sait of compound of formula (I) wherein R is -SMe 4 mg in PP population. (Figure 4) (/-S

Safetv Conclusion:

Overall, magnésium sait of compound of formula (I) wherein R is -SMe 4 mg was safe and well tolerated.

• There were no deaths or SAEs reported in the magnésium sait of compound of formula (I) wherein R îs -SMe 4 mg treatment arm.

• The overall incidence of AEs was Nil.

• There were no persistent changes from baseline in various laboratory parameters. Few events of raised créatinine and five events of raised CPK value were reported during the study. These events were mild and none of these events were consîdered clinically significant by the investigator.

• There was no significant change in weight in magnésium sait of compound of formula (I) wherein R is -SMe 4 mg group in NASH patients at vîsit 2, 3 and 4 visits compared to baseline.

Thus, the compounds of the présent invention and the pharmaceutical composition as described in the spécification are suitable for réduction and removal of lipid accumulated in the liver cells (hépatocytes) for the treatment of Nonalcoholic fatty liver disease (NAFLD) which refers to a wide spectrum of liver diseases ranging from simple fatty liver (steatosis) to nonalcoholic steatohepatitîs (NASH).

Claims (21)

1. We Claim:

   l. A pharmaceutical composition for use in the treatment of nonalcoholic fatty liver disease (NAFLD), the composition comprising (a) the pharmaceutically active substance of Formula (I);

   wherein ‘R’ is selected from hydroxy, hydroxyalkyl, acyl, alkoxy, alkylthio, thioalkyl, aryloxy, and arylthio and M⁺ represents suitable métal cations selected from Na+, K+, Ca+2, and Mg+2;

   (b) suitable additives (c) a suitable stabilizer;

   (d) optionally with one or more pharmaceutically acceptable excipients
2. 2. The pharmaceutical composition according to claim 1 for use în the réduction and removal of lipid accumulated in the liver cells (hépatocytes).
3. 3. The pharmaceutical composition according to claim 1 and 2 for use in preventing and/or reducing/ameliorating the conditions associated with non-alcoholic fatty liver disease (NAFLD) including fatty liver (steatosis), non-alcoholic steatohepatîtis (NASH), and cirrhosis (advanced scarring of the liver)
4. 4. The pharmaceutical composition for use according to claim 3 wherein the disease condition is fatty liver (steatosis).
5. 5. The pharmaceutical composition for use according to claim 3 wherein the disease condition is non-alcoholic steatohepatîtis (NASH).
6. 6. The pharmaceutical composition for use according to claim 3 wherein the disease condition is cirrhosis.
7. 7. The pharmaceutical composition for use according to claim 1 wherein the suitable stabilizer is selected from antioxidants or chelating agents,
8. 8. The phannaceutical composition for use according to claim 7 wherein the suitable antioxidants are selected from citric acid, alpha tocopherol, sodium sulphite, sodium metabisulphite, butylated hydroxy anisole (BHA), BHT (2,6-di-tert-butyl-4methylphénol), monothioglycerol, and Vitamin C (ascorbic acid).
9. 9. The pharmaceutical composition for use according to claim 7 wherein the suitable chelating agents are selected from Disodium EDTA, citric acid and or its salts, maleîc acid, chlorambutol, and chlorhexidine.
10. 10. The pharmaceutical composition for use according to any of claims I to 9 wherein the pharmaceutically active substance of Formula (I) îs used in the range of 0.5 mg to 5 g.
11. 11. The pharmaceutical composition for use according to any of claims 1 to 10 wherein the suitable excipients are selected from solubilizers, diluents, fillers, disintegrants, bînders, lubricants, glidants, wetting agents and solvents.
12. 12. The pharmaceutical composition for use according to any of claims 1 to 11 wherein the suitable additives are selected from sodium benzoate, sodium hydroxide, sodium sulfite and sodium carbonate.
13. 13. The pharmaceutical composition for use according to any of claims 11 to 12 wherein the suitable bînders are selected from acacia alginic acid, tragacanth, carboxymethylcellulose sodium, poly (vinylpyrrolidone), compressible sugar (e.g., NuTab), ethylcellulose, gelatin, liquid glucose, methyl cellulose, povidone and pregelatinized starch, combinations thereof; poly(ethylene glycol), guar gum, polysaccharide, bentonites, sugars, invert sugars, poloxamers (PLURONIC F68, PLURONIC F127), collagen, albumîn, celluloses in nonaqueous solvents, or their suitable combinations; poly(propylene glycol), polyoxyethylene-polypropylene copolymer, polyethylene ester, polyethylene sorbitan ester, poly(ethylene oxide), microcrystalline cellulose, poly(vinylpyrrolidone).
14. 14. The phannaceutical composition for use according to any of claims 11 to 13 wherein the suitable glidants are selected from glicolloîdal silica, calcium silicate, magnésium silicate, silicon hydrogel, corn starch and talc.
15. 15. The phannaceutical composition for use according to any of claims 11 to 13 wherein the suitable glidants are selected from calcium stéarate, magnésium stéarate, minerai oil, stearic acid, and zinc stéarate. Z
16. 16. The pharmaceutical composition for use according to any of claims 11 to 15 wherein the suitable lubricants are selected from calcium stéarate, magnésium stéarate, minerai oil, stearic acid and zinc stéarate.
17. 17. The pharmaceutical composition for use according to any of claims II to 16 wherein the suitable disintegrant are selected from starches such as com starch, potato starch, pre-gelatinîzed and modified starches, sweeteners, clays, microcrystalline cellulose, carsium, alginates, sodium starch glycolate, gums, guar, locust bean, karaya, pectin, and tragacanth.
18. 18. The pharmaceutical composition for use according to any of claims II to 17 wherein the suitable wetting agent are selected from poloxamers, gelatin, casein, Glycerol mono-oleate, lecithin (phosphatîdes), gum acacia, cholestérol, tragacanth, stearic acid, benzalkonium chloride, calcium stéarate, glycerol monostearate, cetostearyl alcohol,sodium lauryl sulphate, sodium dodecyl sulfate, salts of bile acids, cetomacrogol emulsifying wax, sorbitan esters, polyoxyethylene alkyl ethers, polyoxyethylene castor oil dérivatives, polyoxyethylene sorbitan fatty acid esters, polyethylene glycols, polyoxyethylene stéarates, colloïdal silicon dioxide, phosphates, sodium dodecylsulfate, carboxymethylcellulose calcium, carboxy methylcellulose, sodium methyl cellulose, hydroxyethyl cellulose, hydroxylpropyl cellulose, hydroxy propyl methyl cellulose phthalate, noncrystalline cellulose, magnésium aluminum silicate, triethanolamine, polyvinyl alcohol, and poly vinyl pyrrolidone.
19. 19. The pharmaceutical composition for use according to any of claims 1 to 18 which is formulated in tablet or capsule forms.
20. 20. The pharmaceutical composition for use according to any of claims 1 to 19 wherein the pH is maintained in the range of 6 to 10.
21. 21. The pharmaceutical composition for use according to claim 1 containing the pharmaceutically active substance of Formula (I) wherein R is -SMe and M⁺ is Mg⁺². >

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    Effect on ALT