WO2009060452A2 - Procédés et compositions pour le traitement de cristallisation de cholestérol biliaire et d'affections connexes - Google Patents

Procédés et compositions pour le traitement de cristallisation de cholestérol biliaire et d'affections connexes Download PDF

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WO2009060452A2
WO2009060452A2 PCT/IL2008/001467 IL2008001467W WO2009060452A2 WO 2009060452 A2 WO2009060452 A2 WO 2009060452A2 IL 2008001467 W IL2008001467 W IL 2008001467W WO 2009060452 A2 WO2009060452 A2 WO 2009060452A2
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scdl
inhibitor
enzymatic activity
subject
another embodiment
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WO2009060452A3 (fr
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Tuvia Gilat
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Galmed International Ltd
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Galmed International Ltd
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
    • A61K31/19—Carboxylic acids, e.g. valproic acid
    • A61K31/20—Carboxylic acids, e.g. valproic acid having a carboxyl group bound to a chain of seven or more carbon atoms, e.g. stearic, palmitic, arachidic acids
    • A61K31/201—Carboxylic acids, e.g. valproic acid having a carboxyl group bound to a chain of seven or more carbon atoms, e.g. stearic, palmitic, arachidic acids having one or two double bonds, e.g. oleic, linoleic acids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/425—Thiazoles
    • A61K31/426—1,3-Thiazoles
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/425—Thiazoles
    • A61K31/427—Thiazoles not condensed and containing further heterocyclic rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/4427—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems
    • A61K31/4439—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems containing a five-membered ring with nitrogen as a ring hetero atom, e.g. omeprazole
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/56—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids
    • A61K31/575—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids substituted in position 17 beta by a chain of three or more carbon atoms, e.g. cholane, cholestane, ergosterol, sitosterol
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/22—Hormones
    • A61K38/2264—Obesity-gene products, e.g. leptin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics

Definitions

  • the present invention is directed to the field of treating conditions related to biliary cholesterol crystallization, particularly to the administration of inhibitors of Stearoyl Coenzyme A Desaturase 1, other than fatty acid bile acid conjugates.
  • Gallstones (also known as "cholelithiasis”) chiefly cholesterol gallstones, are found in about 15% of the general population in most industrialized countries. Gallstones are concretions that form in the biliary tract, usually in the gallbladder. Their development is insidious, and they may remain asymptomatic for decades. Migration of gallstones may lead to occlusion of the biliary and pancreatic ducts, causing pain (biliary colic) and producing acute complications, such as acute cholecystitis, ascending cholangitis, or acute pancreatitis. Chronic gallstone disease may lead to fibrosis and loss of function of the gallbladder and predisposes to gallbladder cancer. Excision of the gallbladder (cholecystectomy) to cure gallstone disease is among the most frequently performed abdominal surgical procedures.
  • Gallstone formation occurs because certain substances in bile are present in concentrations that approach the limits of solubility. When bile is concentrated in the gallbladder, it can become supersaturated with these substances, which then precipitate from solution as microscopic crystals. The crystals are trapped in gallbladder mucus, producing gallbladder sludge. Over time, the crystals grow, aggregate, and fuse to form macroscopic stones. Occlusion of the ducts by sludge and stones produces the complications of gallstone disease. The only approved medical therapy for gallstone dissolution to date is the oral administration of the bile acid chenodeoxycholic acid (Cheno) or of its epimer ursodeoxycholic acid (Urso).
  • Cheno chenodeoxycholic acid
  • Urso epimer ursodeoxycholic acid
  • bile acid pool that is predominantly composed of Cheno or Urso, respectively.
  • Human bile which is usually supersaturated with cholesterol (which leads to crystallization of biliary cholesterol), becomes under-saturated with cholesterol.
  • the crystallized cholesterol in the gallstone(s) tends to dissolve in the under-saturated bile, and over a period of months or years some of the gallstones dissolve. This therapy is now little used because of its low efficacy.
  • FABACs Fatty Acid Bile Acid Conjugates
  • BAFACs Fatty Acid Bile Acid Conjugates
  • SCD 1 Stearoyl-CoA desaturase 1
  • SCDl catalyzes the rate-limiting reaction of monounsaturated fatty acid (MUFA) synthesis and plays an important role in the synthesis and degradation of fatty acids, the formation of triglycerides and the development of obesity.
  • MUFA monounsaturated fatty acid
  • a variety of SCDl inhibitors are known in the art, including conjugated linoleic acid isomers, oxadiazole pyridazine compounds, 1 -(4-phenoxypiperidin- 1 -yl)-2-arylaminoethanone compounds, cyclopropenoid fatty acids, and thia-fatty acids.
  • conjugated linoleic acid isomers including conjugated linoleic acid isomers, oxadiazole pyridazine compounds, 1 -(4-phenoxypiperidin- 1 -yl)-2-arylaminoethanone compounds, cyclopropenoid fatty acids, and thia-fatty
  • SCDl inhibitors are capable of exerting any effect on 16:1/16:0 and 18:1/18:0 fatty acid ratios in biliary phospholipids. Further, it has not been not shown or suggested that SCDl inhibitors are capable of exerting any effect on the lithogenicity of bile or solubilization and crystallization of biliary cholesteroL Conjugated linoleic acid isomers are well known in the art, and are described, for example, in United States Patents 5,760,083 and 5,770,247 and United States Patent Applications 2001/0031308, 2002/0049346, 2003/0225295, 2004/0018225, 2004/0087512, and 2006/0041017
  • Cholestasis is a condition where bile cannot flow freely from the liver to the duodenum.
  • the two basic types are obstructive cholestasis, due to a mechanical blockage in the duct system (e.g. from a gallstone or malignancy), and metabolic (intrahepatic) cholestasis, a disturbance in bile formation or secretion, due to various causes.
  • PSC Primary sclerosing cholangitis
  • PFIC Progressive familial intrahepatic cholestasis
  • High-GGT PFIC familial intrahepatic cholestasis but with high serum GGT have a condition termed "high-GGT PFIC.” These patients manifest severe progressive intrahepatic cholestasis in the first year and progress toward hepatic failure in the first few years of life. Liver biopsy results reveal expanded portal areas with proliferation of interlobular bile ducts plugged with bile, suggesting an obstructive disorder rather than a primary defect in bile formation. In type 3 PFIC, cholesterol crystallization and stones often occur. There remains an unmet medical need for new therapeutic modes of treating biliary cholesterol crystallization and related conditions (e.g. gallstones of the cholesterol type, PSC, PFIC, obstructive cholestasis, presence of biliary sludge, and cholesterolosis of gallbladder).
  • gallstones of the cholesterol type, PSC, PFIC obstructive cholestasis, presence of biliary sludge
  • the present invention relates to the field of treatment of gallstones, biliary sludge, obstructive cholestasis, and primary sclerosing cholangitis (PSC), and other conditions associated with the precipitation of cholesterol in the biliary tree or liver, particularly comprising administration to a subject of a non-FABAC inhibitor of Stearoyl Coenzyme A Desaturase 1 (SCDl) enzymatic activity or a compound or composition that decreases SCDl expression
  • SCDl Stearoyl Coenzyme A Desaturase 1
  • the present invention provides a method for treating a gallstone in a subject in need thereof, the method comprising the step of administering to the subject an inhibitor of Stearoyl Coenzyme A Desaturase 1 (SCDl) enzymatic activity, wherein the inhibitor of SCDl enzymatic activity is not a fatty acid bile acid conjugate (FABAC), thereby treating gallstones in a subject in need thereof.
  • SCDl enzymatic activity that is inhibited is a hepatic SCDl enzymatic activity.
  • SCDl enzymatic activity in general is inhibited in the subject.
  • the gallstone is a cholesterol gallstone.
  • the present invention provides a method of treating gallstones in general.
  • gallstones in general As is known in the art, over 80% of gallstones in a typical patient population contain cholesterol as their major component. Each possibility represents a separate embodiment of the present invention.
  • non-FABAC SCDl inhibitors markedly decrease gallstone formation.
  • the present invention provides a method for treating a chronic cholestatic disease in a subject in need thereof, the method comprising the step of administering to the subject an inhibitor of SCDl enzymatic activity, wherein the inhibitor of SCDl enzymatic activity is not a fatty acid bile acid conjugate (FABAC), thereby treating a chronic cholestatic disease in a subject in need thereof.
  • the SCDl enzymatic activity that is inhibited is a hepatic SCDl enzymatic activity.
  • SCDl enzymatic activity in general is inhibited in the subject.
  • the chronic cholestatic disease is a cholestasis in the liver.
  • the chronic cholestatic disease is a cholestasis in the biliary tree. In another embodiment, the chronic cholestatic disease is a cholestasis in the liver and biliary tree. In another embodiment, the chronic cholestatic disease predisposes the subject to gallstone formation. In another embodiment, the chronic cholestatic disease is an intrahepatic cholestatic disease. In another embodiment, the chronic cholestatic disease is an extrahepatic cholestatic disease. In another embodiment, the chronic cholestatic disease is an intrahepatic/extrahepatic cholestatic disease. In another embodiment, the chronic cholestatic disease is progressive familial intrahepatic cholestasis.
  • the progressive familial intrahepatic cholestasis is progressive familial intrahepatic cholestasis type 3.
  • the chronic cholestatic disease is another type of chronic cholestatic disease.
  • the present invention provides a method for reducing biliary sludge in a subject in need thereof, the method comprising the step of administering to the subject an inhibitor of SCDl enzymatic activity, wherein the inhibitor of SCDl enzymatic activity is not a FABAC, thereby reducing biliary sludge in a subject in need thereof.
  • the SCDl enzymatic activity that is inhibited is a hepatic SCDl enzymatic activity.
  • SCDl enzymatic activity in general is inhibited in the subject.
  • the present invention provides a method for inhibiting formation of biliary sludge in a subject in need thereof, the method comprising the step of administering to the subject an inhibitor of SCDl enzymatic activity, wherein the inhibitor of SCDl enzymatic activity is not a FABAC, thereby inhibiting formation of biliary sludge in a subject in need thereof.
  • the SCDl enzymatic activity that is inhibited is a hepatic SCDl enzymatic activity.
  • SCDl enzymatic activity in general is inhibited in the subject.
  • the present invention provides a method for treating cholesterolosis of the gallbladder in a subject in need thereof, the method comprising the step of administering to the subject an inhibitor of SCDl enzymatic activity, wherein the inhibitor of SCDl enzymatic activity is not a FABAC, thereby treating cholesterolosis of the gallbladder in a subject in need thereof.
  • the SCDl enzymatic activity that is inhibited is a hepatic SCDl enzymatic activity.
  • SCDl enzymatic activity in general is inhibited in the subject.
  • the present invention provides a method of treating primary sclerosing cholangitis (PSC) in a subject in need thereof, the method comprising the step of administering to the subject an inhibitor of SCDl enzymatic activity, wherein the inhibitor of SCDl enzymatic activity is not a FABAC, thereby treating PSC in a subject in need thereof.
  • the SCDl enzymatic activity that is inhibited is a hepatic SCDl enzymatic activity.
  • SCDl enzymatic activity in general is inhibited in the subject.
  • the present invention provides a method of increasing solubility of cholesterol in bile of a subject in need thereof, the method comprising the step of administering to the subject an inhibitor of SCDl enzymatic activity, wherein the inhibitor of SCDl enzymatic activity is not a FABAC, thereby increasing solubility of cholesterol in bile of a subject in need thereof.
  • the SCDl enzymatic activity that is inhibited is a hepatic SCDl enzymatic activity.
  • SCDl enzymatic activity in general is inhibited in the subject.
  • the present invention provides a method of inhibiting cholesterol crystallization in the biliary tree of a subject in need thereof, the method comprising the step of administering to the subject an inhibitor of SCDl enzymatic activity, wherein the inhibitor of SCDl enzymatic activity is not a FABAC, thereby inhibiting cholesterol crystallization in the biliary tree of a subject in need thereof.
  • the SCDl enzymatic activity that is inhibited is a hepatic SCDl enzymatic activity.
  • SCDl enzymatic activity in general is inhibited in the subject.
  • the present invention provides a method of treating or inhibiting a complication of the cholesterol crystallization. Each possibility represents a separate embodiment of the present invention.
  • the present invention provides a method of reducing the lithogenicity of bile of a subject in need thereof, the method comprising the step of administering to the subject an inhibitor of SCDl enzymatic activity, wherein the inhibitor of SCDl enzymatic activity is not a FABAC, thereby reducing the lithogenicity of bile of a subject in need thereof.
  • the SCDl enzymatic activity that is inhibited is a hepatic SCDl enzymatic activity.
  • SCDl enzymatic activity in general is inhibited in the subject.
  • the present invention provides a method of treating or inhibiting a complication of the cholesterol crystallization. Each possibility represents a separate embodiment of the present invention.
  • the present invention provides a method of inhibiting formation of gallstones in a subject in need thereof, the method comprising administering to the subject an inhibitor of SCDl enzymatic activity, wherein the inhibitor of SCDl enzymatic activity is not a FABAC, thereby inhibiting formation of gallstones in a subject in need thereof.
  • the present invention provides a method of impeding progression of a chronic cholestatic disease in a subject in need thereof, the method comprising administering to the subject an inhibitor of SCDl enzymatic activity, wherein the inhibitor of SCDl enzymatic activity is not a FABAC, thereby impeding progression of a chronic cholestatic disease in a subject in need thereof.
  • the present invention provides a method of reversing the progression of a chronic cholestatic disease, or biliary sludge in a subject in need thereof, the method comprising administering to the subject an inhibitor of SCDl enzymatic activity, wherein the inhibitor of SCDl enzymatic activity is not a FABAC, thereby reversing the progression of gallstones, a chronic cholestatic disease, or biliary sludge in a subject in need thereof.
  • the present invention provides a method of reversing the gallstone formation in a subject in need thereof, the method comprising administering to the subject an inhibitor of SCDl enzymatic activity, wherein the inhibitor of SCDl enzymatic activity is not a FABAC, thereby reversing gallstone formation in a subject.
  • the present invention provides a method for treating obstructive cholestasis in a subject in need thereof, the method comprising the step of administering to the subject an inhibitor of SCDl enzymatic activity, wherein the inhibitor of SCDl enzymatic activity is not a FABAC, thereby treating obstructive cholestasis in a subject in need thereof.
  • the SCDl enzymatic activity that is inhibited is a hepatic SCDl enzymatic activity.
  • SCDl enzymatic activity in general is inhibited in the subject.
  • the obstructive cholestasis is progressive familial intrahepatic cholestasis.
  • the progressive familial intrahepatic cholestasis is type 3 progressive familial intrahepatic cholestasis.
  • the obstructive cholestasis is another type of obstructive cholestasis.
  • the present invention provides use of an inhibitor of SCDl enzymatic activity, wherein the inhibitor of SCDl enzymatic activity is not a FABAC, in the preparation of a medicament for treating gallstones, a chronic cholestatic disease, or primary sclerosing cholangitis (PSC); or reducing biliary sludge.
  • the SCDl enzymatic activity that is inhibited is a hepatic SCDl enzymatic activity.
  • SCDl enzymatic activity in general is inhibited in the subject.
  • the present invention provides a method of treating gallstones, a chronic cholestatic disease, or PSC; or reducing biliary sludge in a subject in need thereof, comprising administering to the subject a compound or composition that decreases SCDl expression, thereby treating gallstones, a chronic cholestatic disease, or PSC; or reducing biliary sludge in a subject in need thereof.
  • the SCDl expression that is decreased is a hepatic SCDl enzymatic activity.
  • SCDl expression in general is decrease in the subject.
  • Mice were fed a high fat, lithogenic, diet for 14 days, without (left bar in each series) or with orally administered 150 mg/kg/day FABAC (Aramchol) supplementation (right bar in each series). Again the monounsaturated/saturated FA ratio was markedly decreased following FABAC therapy.
  • FIG. 3 Stearoyl Coenzyme A Desaturase 1 (SCDl) activity in fresh liver microsomes of C57/BL6 mice.
  • Vertical axis: Specific activity pmol 16:l/min/mg protein. Horizontal axis: test solutions. Control, Aramchol, CLA-10t,12c and 9c,l lt at equimolar concentrations. Aramchol inhibited SCDl activity more than the established CLA inhibitor.
  • FIG. 1 Gallstone formation in C57/BL6 mice after 42 days of lithogenic high fat diet (HFD) alone (first bar) or together with Aramchol (A; second bar) or chow supplemented with 1Ot, 12c CLA at concentrations of 0.1% (third bar) or 0.01% (fourth bar) of food (wt/wt).
  • HFD high fat diet
  • A Aramchol
  • 12c CLA concentrations of 0.1%
  • wt/wt 0.01%
  • the present invention relates to the field of treatment of gallstones, biliary sludge, obstructive cholestasis, and primary sclerosing cholangitis (PSC), particularly comprising administration to a subject of a non-FABAC inhibitor of Stearoyl Coenzyme A Desaturase
  • SCDl SCD 1 enzymatic activity or a compound or composition that decreases SCDl expression.
  • the present invention provides a method for treating a gallstone in a subject in need thereof, the method comprising the step of administering to the subject an inhibitor of a Stearoyl Coenzyme A Desaturase 1 (SCDl) enzymatic activity, wherein the inhibitor of SCDl enzymatic activity is not a fatty acid bile acid conjugate (FABAC), thereby treating a gallstone in a subject in need thereof.
  • the SCDl enzymatic activity that is inhibited is a hepatic SCDl enzymatic activity.
  • SCDl enzymatic activity in general is inhibited in the subject.
  • the gallstone is a cholesterol gallstone.
  • the present invention provides a method of treating gallstones in general. As is known in the art, over 80% of gallstones in a typical patient population contain cholesterol as their major component. Each possibility represents a separate embodiment of the present invention.
  • the gallstones treated by a method of the present invention result from primary sclerosing cholangitis (PSC).
  • PSC primary sclerosing cholangitis
  • SCDl inhibitors markedly increase the saturation of biliary phospholipid fatty acids and thus significantly increase the cholesterol-solubilizing capacity of bile.
  • the present invention provides a pharmaceutical composition comprising an inhibitor of an SCDl enzymatic activity, wherein the inhibitor of SCDl enzymatic activity is not a FABAC, for treating a gallstone.
  • the SCDl enzymatic activity that is inhibited is a hepatic SCDl enzymatic activity.
  • SCDl enzymatic activity in general is inhibited in the subject.
  • “Inhibitor of SCDl enzymatic activity” preferably refers to a substance that exerts a net decrease on SCDl enzymatic activity.
  • the term should be understood to include, in another embodiment, partial antagonists and non-classical inhibitors of SCDl.
  • the term encompasses all SCDl modulators capable of decreasing SCDl activity under conditions that will increase the saturation of biliary phospholipid fatty acids. Each possibility represents a separate embodiment of the present invention.
  • the present invention provides a method for treating a chronic cholestatic disease in a subject in need thereof, the method comprising the step of administering to the subject an inhibitor of an SCDl enzymatic activity, wherein the inhibitor of SCDl enzymatic activity is not a FABAC, thereby treating a chronic cholestatic disease in a subject in need thereof.
  • the SCDl enzymatic activity that is inhibited is a hepatic SCDl enzymatic activity.
  • SCDl enzymatic activity in general is inhibited in the subject.
  • the chronic cholestatic disease is a cholestasis in the liver.
  • the chronic cholestatic disease is a cholestasis in the biliary tree. In another embodiment, the chronic cholestatic disease is a cholestasis in the liver and biliary tree. In another embodiment, the chronic cholestatic disease predisposes the subject to gallstone formation. In another embodiment, the chronic cholestatic disease is an intrahepatic cholestatic disease. In another embodiment, the chronic cholestatic disease is an extrahepatic cholestatic disease. In another embodiment, the chronic cholestatic disease is an intrahepatic/extrahepatic cholestatic disease. In another embodiment, the chronic cholestatic disease is progressive familial intrahepatic cholestasis.
  • the progressive familial intrahepatic cholestasis is progressive familial intrahepatic cholestasis type 3.
  • the chronic cholestatic disease is Caroli's Disease.
  • the chronic cholestatic disease is biliary helminthiasis.
  • the chronic cholestatic disease is biliary strictures.
  • the chronic cholestatic disease is choledocholithiasis.
  • the chronic cholestatic disease is another type of chronic cholestatic disease.
  • the present invention provides a pharmaceutical composition comprising an inhibitor of an SCDl enzymatic activity, wherein the inhibitor of SCDl enzymatic activity is not a FABAC, for treating a chronic cholestatic disease.
  • the SCDl enzymatic activity that is inhibited is a hepatic SCDl enzymatic activity.
  • SCDl enzymatic activity in general is inhibited in the subject.
  • the present invention provides a method for reducing biliary sludge in a subject in need thereof, the method comprising the step of administering to the subject an inhibitor of an SCDl enzymatic activity, wherein the inhibitor of SCDl enzymatic activity is not a FABAC, thereby reducing biliary sludge in a subject in need thereof.
  • the SCDl enzymatic activity that is inhibited is a hepatic SCDl enzymatic activity.
  • SCDl enzymatic activity in general is inhibited in the subject.
  • the present invention provides a method for inhibiting formation of biliary sludge in a subject in need thereof, the method comprising the step of administering to the subject an inhibitor of an SCDl enzymatic activity, wherein the inhibitor of SCDl enzymatic activity is not a FABAC, thereby inhibiting formation of biliary sludge in a subject in need thereof.
  • the SCDl enzymatic activity that is inhibited is a hepatic SCDl enzymatic activity.
  • SCDl enzymatic activity in general is inhibited in the subject.
  • the present invention provides a pharmaceutical composition comprising an inhibitor of an SCDl enzymatic activity, wherein the inhibitor of SCDl enzymatic activity is not a FABAC, for reducing biliary sludge.
  • the SCDl enzymatic activity that is inhibited is a hepatic SCDl enzymatic activity.
  • SCDl enzymatic activity in general is inhibited in the subject.
  • the present invention provides a pharmaceutical composition comprising an inhibitor of an SCDl enzymatic activity, wherein the inhibitor of SCDl enzymatic activity is not a FABAC, for inhibiting formation of biliary sludge.
  • the SCDl enzymatic activity that is inhibited is a hepatic SCDl enzymatic activity.
  • SCDl enzymatic activity in general is inhibited in the subject.
  • the present invention provides a method for treating cholesterolosis of the gallbladder in a subject in need thereof, the method comprising the step of administering to the subject an inhibitor of an SCDl enzymatic activity, wherein the inhibitor of SCDl enzymatic activity is not a FABAC, thereby treating cholesterolosis of the gallbladder in a subject in need thereof.
  • the SCDl enzymatic activity that is inhibited is a hepatic SCDl enzymatic activity.
  • SCDl enzymatic activity in general is inhibited in the subject.
  • the present invention provides a pharmaceutical composition comprising an inhibitor of an SCDl enzymatic activity, wherein the inhibitor of SCDl enzymatic activity is not a FABAC, for treating cholesterolosis of the gallbladder.
  • the SCDl enzymatic activity that is inhibited is a hepatic SCDl enzymatic activity.
  • SCDl enzymatic activity in general is inhibited in the subject.
  • the present invention provides a method of treating primary sclerosing cholangitis (PSC) in a subject in need thereof, the method comprising the step of administering to the subject an inhibitor of an SCDl enzymatic activity, wherein the inhibitor of SCDl enzymatic activity is not a FABAC, thereby treating PSC in a subject in need thereof.
  • the SCDl enzymatic activity that is inhibited is a hepatic SCDl enzymatic activity.
  • SCDl enzymatic activity in general is inhibited in the subject.
  • the present invention provides a pharmaceutical composition comprising an inhibitor of an SCDl enzymatic activity, wherein the inhibitor of SCDl enzymatic activity is not a FABAC, for treating PSC.
  • the SCDl enzymatic activity that is inhibited is a hepatic SCDl enzymatic activity.
  • SCDl enzymatic activity in general is inhibited in the subject.
  • the present invention provides a method of increasing solubility of cholesterol in bile of a subject in need thereof, the method comprising the step of administering to the subject an inhibitor of an SCDl enzymatic activity, wherein the inhibitor of SCDl enzymatic activity is not a FABAC, thereby increasing solubility of cholesterol in bile of a subject in need thereof.
  • the SCDl enzymatic activity that is inhibited is a hepatic SCDl enzymatic activity.
  • SCDl enzymatic activity in general is inhibited in the subject.
  • the present invention provides a pharmaceutical composition comprising an inhibitor of an SCDl enzymatic activity, wherein the inhibitor of SCDl enzymatic activity is not a FABAC, for increasing solubility of cholesterol in bile.
  • the SCDl enzymatic activity that is inhibited is a hepatic SCDl enzymatic activity.
  • SCDl enzymatic activity in general is inhibited in the subject.
  • the present invention provides a method of inhibiting cholesterol crystallization in the biliary tree of a subject in need thereof, the method comprising the step of administering to the subject an inhibitor of an SCDl enzymatic activity, wherein the inhibitor of SCDl enzymatic activity is not a FABAC, thereby inhibiting cholesterol crystallization in the biliary tree of a subject in need thereof.
  • the SCDl enzymatic activity that is inhibited is a hepatic SCDl enzymatic activity.
  • SCDl enzymatic activity in general is inhibited in the subject.
  • the present invention provides a method of treating or inhibiting a complication of the cholesterol crystallization. Each possibility represents a separate embodiment of the present invention.
  • the present invention provides a pharmaceutical composition comprising an inhibitor of an SCDl enzymatic activity, wherein the inhibitor of SCDl enzymatic activity is not a FABAC, for inhibiting cholesterol crystallization in the biliary tree.
  • the SCDl enzymatic activity that is inhibited is a hepatic SCDl enzymatic activity.
  • SCDl enzymatic activity in general is inhibited in the subject.
  • the present invention provides a method of reducing the lithogenicity of bile of a subject in need thereof, the method comprising the step of administering to the subject an inhibitor of an SCDl enzymatic activity, wherein the inhibitor of SCDl enzymatic activity is not a FABAC, thereby reducing the lithogenicity of bile of a subject in need thereof.
  • the SCDl enzymatic activity that is inhibited is a hepatic SCDl enzymatic activity.
  • SCDl enzymatic activity in general is inhibited in the subject.
  • the present invention provides a method of treating or inhibiting a complication of the cholesterol crystallization.
  • the present invention provides a pharmaceutical composition comprising an inhibitor of an SCDl enzymatic activity, wherein the inhibitor of SCDl enzymatic activity is not a FABAC, for reducing the lithogenicity of bile.
  • the SCDl enzymatic activity that is inhibited is a hepatic SCDl enzymatic activity.
  • SCDl enzymatic activity in general is inhibited in the subject.
  • the present invention provides a method of inhibiting formation of gallstones in a subject in need thereof, the method comprising administering to the subject an inhibitor of an SCDl enzymatic activity, wherein the inhibitor of SCDl enzymatic activity is not a FABAC, thereby inhibiting formation of gallstones in a subject in need thereof.
  • the present invention provides a method of impeding progression of a chronic cholestatic disease in a subject in need thereof, the method comprising administering to the subject an inhibitor of an SCDl enzymatic activity, wherein the inhibitor of SCDl enzymatic activity is not a FABAC, thereby impeding progression of a chronic cholestatic disease in a subject in need thereof.
  • the present invention provides a method of reversing the progression of gallstones, a chronic cholestatic disease, or biliary sludge in a subject in need thereof, the method comprising administering to the subject an inhibitor of an SCDl enzymatic activity, wherein the inhibitor of SCDl enzymatic activity is not a FABAC, thereby reversing the progression of gallstones, a chronic cholestatic disease, or biliary sludge in a subject in need thereof.
  • the present invention provides a method for treating obstructive cholestasis in a subject in need thereof, the method comprising the step of administering to the subject an inhibitor of an SCDl enzymatic activity, wherein the inhibitor of SCDl enzymatic activity is not a FABAC, thereby treating obstructive cholestasis in a subject in need thereof.
  • the SCDl enzymatic activity that is inhibited is a hepatic SCDl enzymatic activity.
  • SCDl enzymatic activity in general is inhibited in the subject.
  • the obstructive cholestasis is progressive familial intrahepatic cholestasis.
  • the progressive familial intrahepatic cholestasis is type 3 progressive familial intrahepatic cholestasis.
  • the obstructive cholestasis is another type of obstructive cholestasis.
  • the present invention provides a pharmaceutical composition comprising an inhibitor of an SCDl enzymatic activity, wherein the inhibitor of SCDl enzymatic activity is not a FABAC, for treating obstructive cholestasis.
  • the SCDl enzymatic activity that is inhibited is a hepatic SCDl enzymatic activity.
  • SCDl enzymatic activity in general is inhibited in the subject.
  • the present invention provides use of an inhibitor of an
  • FABAC in the preparation of a medicament for treating gallstones, a chronic cholestatic disease, or primary sclerosing cholangitis (PSC); or reducing biliary sludge.
  • the SCDl enzymatic activity that is inhibited is a hepatic SCDl enzymatic activity.
  • SCDl enzymatic activity in general is inhibited in the subject.
  • the present invention provides a method of treating gallstones, a chronic cholestatic disease, or PSC; or reducing biliary sludge in a subject in need thereof, comprising administering to the subject a compound or composition that decreases SCDl expression, thereby treating gallstones, a chronic cholestatic disease, or
  • SCDl expression that is decreased is a hepatic SCDl enzymatic activity.
  • SCDl expression in general is decrease in the subject.
  • the present invention provides use of a pharmaceutical composition comprising a compound that decreases SCDl expression for treating gallstones, a chronic cholestatic disease, or PSC; or reducing biliary sludge.
  • the SCDl expression that is decreased is a hepatic SCDl enzymatic activity.
  • SCDl expression in general is decrease in the subject.
  • the present invention provides use of a compound or composition that decreases SCDl expression in the preparation of a medicament for treating gallstones, a chronic cholestatic disease, or PSC; or reducing biliary sludge.
  • the SCDl expression that is decreased is a hepatic SCDl enzymatic activity.
  • SCDl expression in general is decrease in the subject.
  • bonding members are NH, P, S, and O-ether.
  • the bonding member can be any bonding member that is not substantially deconjugated during digestion and absorption, with the proviso that an ester bond is not suitable (these are easily degraded).
  • Non-limiting examples of bile acids are cholic acid, ursodeoxycholic acid and chenodeoxycholic acid.
  • FABACs are known in the art, and are described, for example, in WO 99/52932, WO 02/083147 and US 6,589,946, the contents of which are incorporated herein by reference, and in Gilat T et al (Fatty acid bile acid conjugates (FABACs)- new molecules for the prevention of cholesterol crystallization in bile. Gut 2001; 48(l):75-9).
  • Other definitions of FABACs specify 14-22 carbon atoms or 18-22 carbon atoms in the fatty acid radical(s).
  • FABACs The general structure of FABACs is set forth in the structure below.
  • a bile acid is conjugated using an amide bond (at position 3) with fatty acids of variable chain length.
  • Aramchol Arachidyl Amido Cholanoic Acid; also known as "3 ⁇ -arachidylamido-7 ⁇ ,12 ⁇ -dihydroxy-5 ⁇ -cholan-24-oic acid").
  • the non-FABAC SCDl enzymatic activity inhibitor of methods and compositions of the present invention is, in another embodiment, a cyclopropenoid fatty acid (CFA).
  • CFA have characteristics between those of saturated and monounsaturated fatty acids, and closer to the latter, while the strained bond angles of the ring confer a unique chemistry, as described by Christie, W. W. (1970) in Cyclopropane and Cyclopropane Fatty Acids. Topics in Lipid Chemistry 1 : 1-49.
  • the cyclopropane fatty acid synthase gene in E. coli has been cloned and sequenced (Grogan et al., J. Bacteriol. 158:286-295 and Wang et al., Biochemistry 31:11020-11028).
  • the CFA is one found in stercula or cotton seeds.
  • the CFA is sterculic acid (8-(2- octyl-cyclopropenyl)octanoic acid) or malvalic acid (7-(2-octyl-cyclopropenyl)heptanoic acid), which are Cl 8 and C16 derivatives of sterculoyl- and malvaloyl-fatty acids, respectively, having cyclopropene rings at their ⁇ 9 position.
  • sterculic acid 8-(2- octyl-cyclopropenyl)octanoic acid
  • malvalic acid (7-(2-octyl-cyclopropenyl)heptanoic acid
  • the SCDl inhibitor is a pharmaceutically acceptable salt of a CFA.
  • Each possibility represents a separate embodiment of the present invention.
  • the SCDl inhibitor is a thia-fatty acid.
  • Thia-fatty acids are well known in the art, and are described, inter alia, in United States Patent Applications
  • the SCDl inhibitor is a pharmaceutically acceptable salt of a thia-fatty acid.
  • the SCDl inhibitor has the following structure:
  • L is a polar aromatic linker
  • the SCDl inhibitor is a derivative of the above pharmacophore.
  • the SCDl inhibitor is a l-(4-phenoxypiperidin- l-yl)-2-arylaminoethanone compound.
  • the l-(4-phenoxypiperidin-l-yl)-2- arylaminoethanone compound has the following structure:
  • R is selected from the group consisting of CONHMe, CONHEt, CO 2 Me, and Cl.
  • the l-(4-phenoxypiperidin-l-yl)-2-arylaminoethanone compound has the following structure:
  • Rl iTY n1' wherein R 1 is a halide, alkyl, or haloalkyl group, and R 2 has one of the following structures:
  • R 2 is N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl
  • R 1 is selected from Cl and CF 3 .
  • the l-(4-phenoxypiperidin-l-yl)-2-arylaminoethanone compound has the following structure:
  • R 1 is a halide, alkyl, or haloalkyl group
  • R 2 is selected from CONHBn
  • R 2 is selected from CONH 2 , CONHMe, CONHEt, and CONH(CH 2 ) 2 OH.
  • R 1 is selected from Cl and CF 3 .
  • the l-(4-phenoxypiperidin-l-yl)-2-arylaminoethanone compound has the following structure:
  • R is selected from CONHBn, CONH 2 , CONHMe, CONHEt, CONH(iBu),
  • R is selected from CONHMe, CONHEt, CO 2 Me, and Cl.
  • the l-(4-phenoxypiperidin-l-yl)-2- arylaminoethanone compound has the following structure:
  • R is selected from 2-chlorophenoxyl, phenoxyl, 2-bromophenoxyl, 2,5- dichlorophenoxyl, 2-chloro-5-fluorophenoxyl, 2,3-difluorophenoxyl, 2-chloro-3,5- difluorophenoxyl, 2-trifluoromethyl-phenylamino, and Bz.
  • R is selected from 2- chlorophenoxyl, 2-bromophenoxyl, 2,5-dichlorophenoxyl, 2-chloro-5-fluorophenoxyl, 2- and chloro-3,5-difluorophenoxyl.
  • the SCDl inhibitor is a piperidyl arylcarboxamide.
  • the structure of the piperidyl arylcarboxamide is:
  • x and y are each independently 1 , 2 or 3; J and K are each independently N or C(R 11 ); L is N or C(R 4 );
  • W is a direct bond, --N(R ⁇ C(O)--, -C(O)N(R 1 )-, -OC(O)N(R 1 )-, - N(R ⁇ C(O)N(R 1 )-, ⁇ O ⁇ , -N(R 1 )-, ⁇ S(O) t - (where t is 0, 1 or 2), -N(R ⁇ S(O) P -- (where p is 1 or 2), -S(O) P N(R 1 )- (where p is 1 or 2), ⁇ C(O) ⁇ , -OS(O) 2 N(R 1 )-, -OC(O)-, - C(O)O-, -N(R ⁇ C(O)O- or ⁇ C(R 1 ) 2 ⁇ ;
  • V is -N(R 1 )-, -N(R ⁇ C(O)--, -C(O)--, -C(O)O-, -C(S)-, -C(O)N(R 1 )-, - S(O) p - (where p is 1 or 2), -S(O) P N(R 1 )- (where p is 1 or 2), or -C(R 10 )H-; each R 1 is independently selected from the group consisting of hydrogen, C 1 -C 12 alkyl, C 2 -C 12 hydroxyalkyl, C 4 -C 12 cycloalkylalkyl and C 7 -C 19 aralkyl; R 2 is selected from the group consisting of C 1 -C 12 alkyl, C 2 -C 12 alkenyl, C 2 -C 12 hydroxyalkyl, C 2 -C 12 hydroxyalkenyl, C 2 -C 12 alkoxyalkyl, C 3
  • R 3 is selected from the group consisting of C 1 -C 12 alkyl, C 2 -Ci 2 alkenyl, C 2 -Cj 2 hydroxyalkyl, C 2 -C 12 hydroxyalkenyl, C 2 -C 12 alkoxyalkyl, C 3 -C 12 cycloalkyl, C 4 -Ci 2 cycloalkylalkyl, aryl, C 7 -C 19 aralkyl, C 3 -Cj 2 heterocyclyl, C 3 -C 12 heterocyclylalkyl, C 1 -C 12 heteroaryl, and C 3 -C 12 heteroarylalkyl; or R 3 is a multi-ring structure having 2 to 4 rings wherein the rings are independently selected from the group consisting of cycloalkyl, heterocyclyl, aryl and heteroaryl and where some or all of the rings may be fused to each other; each R 4 is independently selected from hydrogen, fiuoro, chloro, C 1 -C 12 al
  • R 5 , R 5a , R 6 , R 6a , R 7 , R 7a , R 8 , and R 8a are each independently selected from hydrogen or Ci-C 3 alkyl; or R 5 and R 5a together, R 6 and R 6a together, or R 7 and R 7a together, or R 8 and R 8a together are an oxo group, provided that when V is -C(O)- , R 6 and R 6a together or R 8 and R 8a together do not form an oxo group, while the remaining R 5 , R 5a , R 6 , R 6a , R 7 , R 7a , R 8 , and R 8a are each independently selected from hydrogen or Ci-C 3 alkyl; or one of R 5 , R 5a , R 6 , and R 6a , together with one of R 7 , R 7a , R 8 , R 8a , forms a direct bond or an alkylene bridge, while the remaining R 5 , R 5a , R 6
  • R 11 is independently selected from hydrogen, fiuoro, chloro, Ci-Ci 2 alkyl or Ci-Ci 2 alkoxy.
  • piperidyl arylcarboxamide in another piperidyl arylcarboxamide compound known in the art.
  • Methods for making and using piperidyl arylcarboxamide compounds are well known in the art, and are described, inter alia, in
  • the SCDl inhibitor is an inhibitor described in WO 05/011657 in the name of Abreo, Chafeev, et al.
  • the SCDl inhibitor is an inhibitor described in US 2005/0119251 in the name of Fu, Kodumuru, et al.
  • the SCDl inhibitor is a pyridazine carboxamide.
  • the pyridazine carboxamide has one of the following structures:
  • the SCDl inhibitor is a derivative of one of the above structures.
  • the SCDl inhibitor is a pyridazine isoamyl carboxamide.
  • the pyridazine isoamyl carboxamide has one of the structures 7a- 7b:
  • the pyridazine carboxamide has the following structure:
  • Ar is phenyl, 2-fluorophenyl, 3 -fluorophenyl, 4-fluorophenyl, 2,3- difluorophenyl, 2,4-difluorophenyl, 2,5-difluorophenyl, 2,6-difluorophenyl, 2- trifluoromethylphenyl, 2-chlorophenyl, or 2,5-dichlorophenyl.
  • the pyridazine carboxamide has the structure "15a-q":
  • R 1 is methyl, H, or one of the following structures: Y ⁇ ⁇ x * ⁇ r ⁇ r * Ou
  • Ar is 2-fluorophenyl, 2-chlorophenyl, 2-bromophenyl, 2-cyanophenyl, 2,5- dichlorophenyl, 2-chloro-5-fluorophenyl, or 2-methoxyphenyl.
  • the SCDl inhibitor is an oxadiazole pyridazine.
  • the oxadiazole pyridazine has the structure "20a-e":
  • the oxadiazole pyridazine has the structure "24a-f ':
  • Ar is phenyl, 2-fluorophenyl, 2-chlorophenyl, 2-methylphenyl, or 2- methoxyphenyl, and R 1 is Me or PhCH 2 -.
  • the oxadiazole pyridazine has the structure "28a- c":
  • Ar is 2-chlorophenyl or 2-chloro-5-fluorophenyl, and R 1 is Me or PhCH 2 -.
  • Ar is 2-chloro-5- fluorophenyl
  • R 1 is Me
  • the SCDl inhibitor is a heteroaryl pyridazine.
  • heteroaryl pyridazine has the following structure:
  • Ar is 2-chlorophenyl or 2-chloro-5-fluorophenyl, and Het is one of the following structures:
  • the compound that decreases SCDl activity of the present invention is, in another embodiment, a conjugated linoleic acid isomer.
  • Conjugated linoleic acid isomer refers, in another embodiment, to a linoleic acid isomer containing two double bonds that are conjugated, i.e. separated by a one single bond.
  • the CLA isomer is a 1Ot, 12c linoleic acid isomer having the formula:
  • the CLA isomer is a 10,12 linoleic acid isomer. In another embodiment, the CLA isomer is a 10,12 dienoic linoleic acid isomer. In another embodiment, the CLA isomer is a 1Ot, 12c dienoic linoleic acid isomer. In another embodiment, the CLA isomer is a conjugated dienoic linoleic acid derivative. More preferably, the CLA isomer is a conjugated 10,12 dienoic linoleic acid derivative. In another embodiment, the CLA isomer is a trans- 10 isomer of linoleic acid.
  • the CLA isomer is a cis-10 isomer. In another embodiment, the CLA isomer is the trans- 10,cis- 12 isomer. In another embodiment, the CLA isomer is the cis-10,cis-12 isomer. In another embodiment, the CLA isomer is the cis-10,trans-12 isomer. In another embodiment, the CLA isomer is the trans- 10,trans- 12 isomer.
  • Each possibility represents a separate embodiment of the present invention.
  • the 10,12 linoleic acid isomer is 9,12,15-octadecatrienoic acid ( ⁇ -linolenic acid) [18:3, ⁇ 3]. In another embodiment, the 10,12 linoleic acid isomer is 6,9,12-octadecatrienoic acid ( ⁇ -linolenic acid) [18:3, ⁇ 6]. In another embodiment, the 10,12 linoleic acid isomer is 8,11,14-eicosatrienoic acid (dihomo- ⁇ -linolenic acid) [20:3 co6].
  • the 10,12 linoleic acid isomer is 6,9-octadecadienoic acid [18:2, ⁇ 9]. In another embodiment, the 10,12 linoleic acid isomer is trans8,transl ⁇ ,cisl2- 18:3 linolenic acid. Each possibility represents a separate embodiment of the present invention.
  • the compound that decreases SCDl activity is a linoleic acid derivative having conjugated double bonds.
  • the compound is a linoleic acid derivative having conjugated double bonds at the 10 and 12 positions.
  • the compound is a linoleic acid derivative having conjugated double bonds at the trans- 10,cis- 12 positions.
  • the compound is selected from the group consisting of 9-HODE (9-hydroxy-trans-10,cis-12-octadecodienoic acid) and 9-HOO (9-hydroperoxy-trans-10,cis-12-octadecodienoic acid) (first and second structures below, respectively).
  • the compound is any other linoleic acid derivative known in the art that has conjugated double bonds at the trans- 10,cis- 12 positions. Each possibility represents a separate embodiment of the present invention.
  • the CLA isomer is another CLA isomer known in the art.
  • the SCDl inhibitor is a pharmaceutically acceptable salt of a CLA isomer.
  • Conjugated linoleic acid isomers and their preparation are well known in the art, and are described, for example, in United States Patents 5,892,074, 5,760,083, and 5,770,247 and United States Patent Applications 2001/0031308, 2002/0049346, 2003/0225295, 2004/0018225, 2004/0087512, 2005/0215641, 2006/0041017, 2006/0106238, 2007/0078274, and 2007/0196446, and in Ha YL et al 1990 (Inhibition of benzo(a)pyrene-induced mouse forestomach neoplasia by conjugated dienoic derivatives of linoleic acid. Cancer Res.
  • the SCDl inhibitor is dihomo-gamma linolenic acid (GLLA).
  • GLLA dihomo-gamma linolenic acid
  • the SCDl inhibitor is a pyridazine derivative.
  • Pyridazine derivatives are well known in the art, and are described, inter alia, in United States patent application 2008/0207587, the contents of which are incorporated herein by reference. Each possibility represents a separate embodiment of the present invention.
  • the SCDl inhibitor is an inhibitor described in Yee et al, Compartmentalization of stearoyl-coenzyme A desaturase 1 activity in HepG2 cells, J Lipid Res, 49: 2124-2134, 2008.
  • Each possibility represents a separate embodiment of the present invention.
  • Many strategies are available to lower SCDl protein level. For example, one can increase the degradation rate of the enzyme or inhibit rate of synthesis of the enzyme. The synthesis of the enzyme can be inhibited at transcriptional level or translational level by known genetic techniques. Since SCDl is regulated by several known transcription factors (e.g. PP AR-.
  • any agent that affects the activity of such transcription factors can be used to alter the expression of the SCDl gene at transcriptional level.
  • One group of such agents includes thiazolidine compounds which are known to activate PPAR- gamma and inhibit SCDl transcription. These compounds include Rosiglitazone, Pioglitazone, Ciglitazone, Englitazone, Troglitazone, and BRL49653. Another agent is leptin, which has been shown to inhibit SCDl expression (Cohen, P. et al., Science. 297: 240-243, 2002).
  • the compound or composition that decreases SCDl expression of the present invention is selected from the group consisting of linoleic acid, linolenic acid, arachidonic acid, and eicosapentaenoic acid.
  • the level of SCDl protein is decreased by downregulating
  • PRP 19 precursor RNA processing 19 protein
  • SCDl expression inhibitors are described in Lin J et al, CNS melanocortin and leptin effects on stearoyl-CoA desaturase-1 and resistin expression. Biochem Biophys Res Commun. 2003 Nov 14;311(2):324-8); and Sessler AM et al, Regulation of stearoyl- CoA desaturase 1 mRNA stability by polyunsaturated fatty acids in 3T3-L1 adipocytes. J Biol Chem. 1996 Nov 22;271(47):29854-8), the contents of which are incorporated herein by reference.
  • Gallstones refers to concretions that form in the biliary tract, usually in the gallbladder. Gallstones are also known as “cholelithiasis.” Migration of gallstones may lead to occlusion of the biliary and pancreatic ducts, causing pain (biliary colic) and producing acute complications, such as acute cholecystitis, ascending cholangitis, or acute pancreatitis. Chronic gallstone disease may lead to fibrosis and loss of function of the gallbladder and predisposes to gallbladder cancer. It will be understood to one skilled in the art that the present invention encompasses methods for treatment and alleviation of these and other complications of gallstones.
  • PFIC Progressive familial intrahepatic cholestasis
  • PFIC-I the former Byler disease
  • PFIC-2 the former Byler disease
  • PFIC-I and PFIC-2 have few clinical differences, and both are caused by the absence of a gene product function for canalicular export and bile formation.
  • PFIC Progressive familial intrahepatic cholestasis
  • Patients with familial intrahepatic cholestasis but with high serum GGT have a condition termed "high-GGT PFIC.” These patients manifest severe progressive intrahepatic cholestasis in the first year and progress toward hepatic failure in the first few years of life. Liver biopsy results reveal expanded portal areas with proliferation of interlobular bile ducts plugged with bile, indicating an obstructive disorder rather than a primary defect in bile formation.
  • Cholesterolosis refers to a condition in which cholesterol is deposited in tissues in abnormal quantities.
  • Cholestasis refers to a pathologic state of reduced bile formation or flow, having intrahepatic or extrahepatic causes, wherein substances normally excreted into bile are retained. The serum concentrations of conjugated bilirubin and bile salts may be measured.
  • Primary sclerosing cholangitis (PSC), refers to a chronic cholestatic liver disease that is recognized increasingly in children. The diagnosis is based on a combination of clinical features and cholestatic biochemical profile, along with typical cholangiographic abnormalities, and confirmed by liver histology findings.
  • PSC intrahepatic and extrahepatic biliary tree characterize PSC.
  • PSC is usually progressive, leading to cirrhosis, portal hypertension, and liver failure. Gallstones can form above strictured areas.
  • Effective medical treatment modalities for childhood PSC are undetermined.
  • Liver transplantation remains the only effective therapeutic option for patients with end-stage liver disease from PSC. Measurement of SCDl enzymatic activity
  • the assay employs a microsomal assay having a measurable SCDl biological activity.
  • a suitable assay may be taken by modifying and scaling up the rat liver microsomal assay essentially as described by Shimomura et al. (Shimomura, L, Shimano, H., Korn, B. S., Bashmakov, Y., and Horton, J. D. (1998). Tissues are homogenized in 10 vol. of buffer A (0.1 M potassium buffer, pH 7.4).
  • microsomal membrane fractions (100,000 X-g pellet) are isolated by sequential centrifugation. Reactions are performed at 37° C. for 5-20 min with 100 ⁇ g of protein homogenate and 60 ⁇ M of [l-14C]-stearoyl- CoA (60,000 dpm), 2 mM of NADH, 0. IM of Tris/HCl buffer (pH 7.2). After the reaction, fatty acids are extracted and then methylated with 10% acetic chloride/methanol. Saturated fatty acid and monounsaturated fatty acid methyl esters are separated by 10% AgNO 3 - impregnated TLC using hexane/diethyl ether (9:1) as developing solution.
  • the plates are sprayed with 0.2% 2', 7'-dichlorofluorescein in 95% ethanol and the lipids are identified under UV light.
  • the fractions are scraped off the plate, and the radioactivity is measured using a liquid scintillation counter.
  • Specific embodiments of such SCDl biological activity assay take advantage of the fact that the SCD reaction produces, in addition to the monounsaturated fatty acyl-CoA product, H 2 O. If 3 H is introduced into the C-9 and C-IO positions of the fatty-acyl-CoA substrate, then some of the radioactive protons from this reaction will end up in water. Thus, the measurement of the activity would involve the measurement of radioactive water.
  • substrates such as charcoal, hydrophobic beads, or ordinary solvents in acid pH.
  • a screening assay is utilized to identify new SCDl inhibitors.
  • Methods for measuring SCDl activity are well known in the art, and are described, inter alia, in Leikin and Brenner, Cholesterol induced microsomal changes modulate desaturase activities. Biochim.Biophys.Acta 922: 294-303, 1987.
  • the SCDl enzyme catalyzes the delta 9 mono-unsaturation of fatty acids, e.g. Palmitic Acid (16:0) to Palmitoleic Acid (16:1) and stearic acid (18:0) to 18:1.
  • screening assays measure SCDl biological activity indirectly.
  • Standard high-throughput screening assays center on ligand-receptor assays. These may be fluorescence-based or luminescence-based or may utilize radiolabel detection.
  • Enzyme immunoassays can be run on a wide variety of formats for identifying compounds that interact with SCDl proteins. These assays may employ prompt fluorescence or time- resolved fluorescence immunoassays which are well known.
  • 32 P-labeled ATP is typically used for protein kinase assays. Phosphorylated products can be separated for counting by a variety of methods. Scintillation proximity assay technology is an enhanced method of radiolabel assay.
  • Cell-based desaturation assays can be used to track SCDl activity levels. By tracking the conversion of stearate to oleate in cells (or palimitic acid to palmitoleic acid) (3T3L1 adipocytes are known to have high SCDl expression and readily take up stearate when complexed to BSA) one can evaluate compounds found to be inhibitory in the primary screen for additional qualities or characteristics such as whether they are cell permeable, lethal to cells, and/or competent to inhibit SCDl activity in cells.
  • This cell- based assay may employ a recombinant cell line containing a delta-9 desaturase. The recombinant gene is optionally under control of an inducible promoter and the cell line preferably over-expresses SCDl protein.
  • Cell-based assays may be preferred, for they leave the SCDl gene in its native format.
  • Particularly promising for SCDl analysis in these types of assays are fluorescence polarization assays. The extent to which light remains polarized depends on the degree to which the tag has rotated in the time interval between excitation and emission. Since the measurement is sensitive to the tumbling rate of molecules, it can be used to measure changes in membrane fluidity characteristics that are induced by SCDl activity—namely the delta-9 desaturation activity of the cell.
  • An alternate assay for SCDl involves a FRET assay. FRET assays measure fluorescence resonance energy transfer which occurs between a fluorescent molecule donor and an acceptor, or quencher. Such an assay may be suitable to measure changes in membrane fluidity or temperature sensitivity characteristics induced by SCDl biological activity.
  • screening assays of the present invention are conducted using high-throughput robotic systems.
  • Such systems may include chip devices developed by, among others, Caliper, Inc., ACLARA BioSciences, Cellomics, Inc., Aurora Biosciences Inc., and others.
  • Preferred assays are readily adapted to the format used for drug screening, which may consist of a multi-well (e.g., 96-well, 384-well or 1536-well or greater) format. Modification of the assay to optimize it for drug screening would include scaling down and streamlining the procedure, modifying the labeling method, altering the incubation time, and changing the method of calculating SCDl biological activity and so on. In all these cases, the SCDl biological activity assay remains conceptually the same, though experimental modifications may be made.
  • An SCDl activity assay may also be carried out as a cell-free assay employing a cellular fractional, such as a microsomal fraction, obtained by conventional methods of differential cellular fractionation, most commonly by ultracentrifugation methods.
  • the screening assay may employ a vector construct comprising a SCDl promoter sequence operably linked to a reporter gene.
  • a vector construct comprising a SCDl promoter sequence operably linked to a reporter gene.
  • the assay makes use of H-stearoyl CoA (with the H on the 9 and 10 carbon atoms), the substrate for SCDl. Desaturation by SCDl, produces oleoyl CoA and 3 H-water molecules. The reaction is run at room temperature, quenched with acid and then activated charcoal is used to separate unreacted substrate from the radioactive water product. The charcoal is sedimented, and the amount of radioactivity in the supernatant is determined by liquid scintillation counting. This assay is specific for
  • the method is easily adapted to high throughput as it is cell-free, conducted at room temperature and is relatively brief (1 hour reaction time period versus previous period of 2 days.
  • Bile is devoid of triglycerides and has little free fatty acids. Thus, almost all biliary fatty acids are constituents of phospholipids, the most abundant by far being Phosphatidylcholine (PC).
  • PC Phosphatidylcholine
  • SCDl refers to a protein also referred to as “stearoyl-CoA desaturase” and “delta-9-desaturase.”
  • amino acid sequence of the SCDl protein is as set forth in SEQ ID NO: 1 :
  • the SCDl is a homologue of SEQ ID NO: 1. In another embodiment, the SCDl is a variant of SEQ ID NO: 1. In another embodiment, the SCDl is a fragment of SEQ ID NO: 1. In another embodiment, the SCDl is a homologue of a fragment of SEQ ID NO: 1. In another embodiment, the SCDl is a variant of a fragment of SEQ ID NO: 1. Each possibility represents a separate embodiment of the present invention.
  • the SCDl protein is encoded by a nucleotide molecule with a sequence set forth in SEQ ID NO: 2: ggcaggacgaggtggcaccaaattcccttcggccaatgacgagccggagtttacagaagcctcattagcatttccccagaggcag gggcaggggcagaggccgggtggtgtggtgtcggtgtcggtgtcggcagcatcccccctgctgcggtcgcgcgcgagcctcggcggcgcgagcctcggccccgcgagcctcggccccgcgagcctcggccccccccccccccccttacctccacgcgggaccgcccgcgccagtcaactcctcgccctgcttggcag
  • the SCDl is a homologue of SEQ ID NO: 2. In another embodiment, the SCDl is a variant of SEQ ID NO: 2. In another embodiment, the SCDl is a fragment of SEQ ID NO: 2. In another embodiment, the SCDl is a homologue of a fragment of SEQ ID NO: 2. In another embodiment, the SCDl is a variant of a fragment of SEQ ID NO: 2. Each possibility represents a separate embodiment of the present invention.
  • the DTA is at least 60% homologous to a sequence selected from SEQ ID NO: 1 and NO: 2. In another embodiment, the DTA is at least 65% homologous to SEQ ID NO: 1 or NO: 2. In another embodiment, the DTA is at least 70% homologous to SEQ ID NO: 1 or NO: 2. In another embodiment, the DTA is at least 72% homologous to SEQ ID NO: 1 or NO: 2. In another embodiment, the DTA is at least 74% homologous to SEQ ID NO: 1 or NO: 2. In another embodiment, the DTA is at least 76% homologous to SEQ ID NO: 1 or NO: 2. In another embodiment, the DTA is at least 78% homologous to SEQ ID NO: 1 or NO: 2.
  • the DTA is at least 80% homologous to SEQ ID NO: 1 or NO: 2. In another embodiment, the DTA is at least 82% homologous to SEQ ID NO: 1 or NO: 2. In another embodiment, the DTA is at least 84% homologous to SEQ ID NO: 1 or NO: 2. In another embodiment, the DTA is at least 86% homologous to SEQ ID NO: 1 or NO: 2. In another embodiment, the DTA is at least 88% homologous to SEQ ID NO: 1 or NO: 2. In another embodiment, the DTA is at least 90% homologous to SEQ ID NO: 1 or NO: 2. In another embodiment, the DTA is at least 92% homologous to SEQ ID NO: 1 or NO: 2.
  • the DTA is at least 94% homologous to SEQ ID NO: 1 or NO: 2. In another embodiment, the DTA is at least 95% homologous to SEQ ID NO: 1 or NO: 2. In another embodiment, the DTA is at least 96% homologous to SEQ ID NO: 1 or NO: 2. In another embodiment, the DTA is at least 97% homologous to SEQ ID NO: 1 or NO: 2. In another embodiment, the DTA is at least 98% homologous to SEQ ID NO: 1 or NO: 2. In another embodiment, the DTA is at least 99% homologous to SEQ ID NO: 1 or NO: 2. In another embodiment, the DTA is over 99% homologous to SEQ ID NO: 1 or NO: 2.
  • variant refers to a pharmaceutically acceptable salt, homologue, analogue, or fragment of a nucleotide sequence useful for the invention (e.g., vector sequences, transcriptional regulatory sequences, cloned polynucleotides of interest, etc.)- Encompassed within the term “variant” are chemically modified natural and synthetic nucleotide molecules. Also encompassed within the term “variant” are conservative substitutions within the nucleotide sequence of the molecule. In addition, non-conservative substitutions within the nucleotide sequence of the molecule are encompassed within the term "variant” as used herein.
  • a transcriptional regulatory sequence for example, a promoter or enhancer
  • a transcriptional regulatory sequence can be the complete native regulatory sequence of the gene, a portion of the native regulatory sequence, a chimeric construction of the native regulatory sequence, a combinatorial construction of one or more native regulatory sequences, or a variant of the native regulatory sequence obtained by, for example, deletion, addition or replacement of at least one nucleotide.
  • a variant regulatory sequence can comprise modified nucleotides.
  • the derived sequence preferably demonstrates properties of control/regulation (e.g., increase/decrease) of the expression of coding sequences operably linked thereto.
  • Alterations in regulatory sequences of the present invention can be generated using a variety of chemical and enzymatic methods which are well known to those skilled in the art. For example, regions of the sequences defined by restriction sites can be deleted, oligonucleotide-directed mutagenesis can be employed to alter the sequence in a defined way and/or to introduce restriction sites in specific regions within the sequence. Additionally, deletion mutants can be generated using DNA nucleases such as Bal31 or ExoIII and Sl nuclease. Progressively larger deletions in the regulatory sequences are generated by incubating the DNA with nucleases for increased periods of time.
  • the compounds or compositions of the invention are provided in packs in a form ready for administration.
  • the compounds or compositions are provided in concentrated form in packs, optionally with the diluent required to make final solution(s) for administration.
  • the product contains a compound useful in the invention in solid form and, optionally, a separate container with a suitable solvent or carrier for the compound useful in the invention.
  • the above packs/kits include other components, e.g., instructions for dilution, mixing and/or administration of the product, other containers, syringes, needles, etc. Other such pack/kit components will be readily apparent to one of skill in the art.
  • the kits further comprise instructions for administering the compound or composition of the invention.
  • a "pharmaceutical composition” refers to a preparation of one or more of the active ingredients described herein, with other components such as physiologically suitable carriers and excipients.
  • the purpose of a pharmaceutical composition is to facilitate administration of a compound to a subject.
  • a therapeutic composition refers to a carrier or a diluent that does not cause significant irritation to an organism and does not abrogate the biological activity and properties of the administered compound.
  • excipient refers to an inert substance added to a pharmaceutical composition to further facilitate administration of an active ingredient. Examples, without limitation, of excipients particularly suitable include calcium carbonate, calcium phosphate, various sugars and types of starch, cellulose derivatives, gelatin, vegetable oils, and polyethylene glycols.
  • a therapeutic composition further comprises a pharmaceutically acceptable carrier.
  • a “carrier” refers to any substance suitable as a vehicle for delivering a therapeutic agent to a suitable in vivo or in vitro site.
  • carriers can act as a pharmaceutically acceptable excipient of a therapeutic composition of the present invention.
  • non-targeting carriers include, but are not limited to water, phosphate buffered saline, Ringer's solution, dextrose solution, serum-containing solutions, Hank's solution, other aqueous physiologically balanced solutions, oils, esters and glycols.
  • Aqueous carriers can contain suitable auxiliary substances required to approximate the physiological conditions of the recipient, for example, by enhancing chemical stability and isotonicity.
  • Suitable auxiliary substances include, for example, sodium acetate, sodium chloride, sodium lactate, potassium chloride, calcium chloride, and other substances used to produce phosphate buffer, Tris buffer, and bicarbonate buffer.
  • Auxiliary substances can also include preservatives, such as thimerosal, m- and o-cresol, formalin and benzol alcohol.
  • Preferred auxiliary substances for aerosol delivery include surfactant substances non-toxic to a subject, for example, esters or partial esters of fatty acids containing from about six to about twenty-two carbon atoms.
  • Other carriers can include metal particles (e.g., gold particles) for use with, for example, a biolistic gun through the skin.
  • Therapeutic compositions of the present invention can be sterilized by conventional methods.
  • Targeting carriers are herein referred to as "delivery vehicles.”
  • Delivery vehicles of the present invention are capable of delivering a therapeutic compound of the present invention to a target site in a subject.
  • a "target site” refers to a site in a subject to which one desires to deliver a therapeutic composition.
  • Examples of delivery vehicles particularly suitable include, but are not limited to, artificial and natural lipid-containing delivery vehicles. Natural lipid-containing delivery vehicles include cells and cellular membranes. Artificial lipid-containing delivery vehicles include liposomes and micelles.
  • a delivery vehicle of the present invention can be modified to target to a particular site in a subject, thereby targeting and making use of a therapeutic compound of the present invention at that site.
  • Suitable modifications include manipulating the chemical formula of the lipid portion of the delivery vehicle and/or introducing into the vehicle a compound capable of specifically targeting a delivery vehicle to a preferred site, for example, a preferred cell type.
  • Specifically targeting refers to causing a delivery vehicle to bind to a particular cell by the interaction of the compound in the vehicle to a molecule on the surface of the cell.
  • Suitable targeting compounds include ligands capable of selectively (i.e., specifically) binding another molecule at a particular site. Examples of such ligands include antibodies, antigens, receptors and receptor ligands.
  • an antibody specific for an antigen found on the surface of a target cell can be introduced to the outer surface of a liposome delivery vehicle so as to target the delivery vehicle to the target cell.
  • Manipulating the chemical formula of the lipid portion of the delivery vehicle can modulate the extracellular or intracellular targeting of the delivery vehicle.
  • a chemical can be added to the lipid formula of a liposome that alters the charge of the lipid bilayer of the liposome so that the liposome fuses with particular cells having particular charge characteristics.
  • a delivery vehicle particularly suitable is a liposome.
  • a liposome is capable of remaining stable in a subject for a sufficient amount of time to deliver a therapeutic compound of the present invention to a preferred site in the subject.
  • a liposome of the present invention is preferably stable in the subject into which it has been administered for at least about 30 minutes, more preferably for at least about 1 hour and even more preferably for at least about 24 hours.
  • Suitable liposomes for use with the present invention include any liposome.
  • Preferred liposomes of the present invention include those liposomes standardly used in, for example, methods known to those of skill in the art.
  • more preferred liposomes comprise liposomes having a polycationic lipid composition and/or liposomes having a cholesterol backbone conjugated to polyethylene glycol.
  • terapéuticaally effective amounts means that the compounds or compositions of the invention are administered to the subject in amounts that are effective to treat that subject.
  • An effective administration protocol (i.e., administering a pharmaceutical composition in an effective manner) comprises suitable dose parameters and modes of administration that result in treatment of a disease.
  • Effective dose parameters and modes of administration can be determined using methods standard in the art for a particular disease. Such methods include, for example, determination of survival rates, side effects (i.e., toxicity) and progression or regression of disease.
  • a suitable single dose size is a dose that is capable of treating a subject with disease when administered one or more times over a suitable time period.
  • compositions of the present invention are administered orally.
  • Other suitable routes of administration may, for example, include oral, rectal, transmucosal, especially transnasal, intestinal or parenteral delivery.
  • Solutions or suspensions used for parenteral, intradermal, or subcutaneous application can include the following components: a sterile diluent such as water for injection, saline solution, fixed oils, polyethylene glycols, glycerine, propylene glycol (or other synthetic solvents), antibacterial agents (e.g., benzyl alcohol, methyl parabens), antioxidants (e.g., ascorbic acid, sodium bisulfite), chelating agents (e.g., ethylenediaminetetraacetic acid), buffers (e.g., acetates, citrates, phosphates), and agents that adjust tonicity (e.g., sodium chloride, dextrose).
  • the pH can be adjusted with acids or bases, such as hydrochloric acid or sodium hydroxide, for example.
  • the parenteral preparation can be enclosed in ampules, disposable syringes or multiple dose vials made of glass or plastic.
  • compositions adapted for parenteral administration include, but are not limited to, aqueous and non-aqueous sterile injectable solutions or suspensions, which can contain antioxidants, buffers, bacteriostats and solutes that render the compositions substantially isotonic with the blood of an intended recipient.
  • Such compositions can also comprise water, alcohols, polyols, glycerine and vegetable oils, for example.
  • Extemporaneous injection solutions and suspensions can be prepared from sterile powders, granules and tablets.
  • Such compositions should comprise a therapeutically effective amount of a vector of the invention and/or other therapeutic agent, together with a suitable amount of carrier so as to provide the form for proper administration to the subject.
  • the formulation should suit the mode of administration.
  • Wild type C57/BL6/J mice were administered a High-Fat Diet (HFD), (containing 1% cholesterol, 0.5% cholic acid, 15% butter and 2% corn oil) over 10 days.
  • HFD High-Fat Diet
  • Half of the animals in each group received FABAC (Aramchol, 150 mg/kg/day) by gavage, while the other half received a gavage with the same volume of vehicle (saline).
  • FABAC Amphos, 150 mg/kg/day
  • mice were sacrificed under anesthesia, and bile was extracted from the gallbladder. Bile from mice receiving each treatment were pooled. Palmitic and palmitoleic phosphatidylcholine fatty acids were isolated and analyzed by gas chromatograph (GC).
  • GC gas chromatograph
  • the second fraction was applied to thin-layer chromatography (TLC) plates.
  • Bile phospholipids were separated with the developing solvent, chloroform:methanol:acetic acid:DDW, (50:30:8:4,v:v:v:v). Once separated and identified, individual phospholipids were extracted twice from the silica gel with 5 ml of the solvent mixture: chloroform:methanol:DDW (5:5:1). The two extracts were combined, 5 ml DDW was added, the mixture was vortexed and centrifuged at 5000 rpm for 10 min, and the upper water phase was discarded. The lower phase was used for esterification and measurement of the fatty acids of each individual phospholipid.
  • Fatty Acid Bile Acid Conjugates Fatty Acid Bile Acid Conjugates (FABACs) on the saturation of biliary phospholipid fatty acids was tested.
  • Rodents were administered a High-Fat Diet (HFD), with or without oral administration of Aramchol (arachidyl amido cholanoic acid; also known as "3 ⁇ -arachidylamido-7 ⁇ ,12 ⁇ -dehydroxy-5 ⁇ -cholan-24-oic acid”) over the same time period.
  • Aramchol significantly increased 16:0 saturated fatty acid levels, decreased 16:1 monounsaturated fatty acid levels (Figure IA), and reduced the 16:1/16:0 fatty acid ratio ( Figure IB).
  • FABACs markedly increase saturation of biliary phospholipids.
  • AA Liver microsomal suspensions of 20-30 mg/ml protein were prepared from the experimental mice.
  • 0.25 ml Incubation Solution (IS) was prepared (Table 1 below sets forth the composition per assay point) and placed in chilled 2.5ml Eppendorf tubes, and lO ⁇ l propylene glycol, containing 16.6 nmol/0.25 ⁇ Ci/tube substrate 14 C palmitic acid, was added. The volume was brought up to 0.5 ml with DDW, and tubes were vortexed and transferred to a 37°C incubation bath. 1.6 mg microsomes/ml in up to 80 ⁇ l was added.
  • the substrate/inhibitor ratio was 4.7.
  • the assay was run for 20 min, after which it was stopped with 0.5ml 10% KOH in Ethanol.
  • TLC plates were impregnated with 10% AgNO 3 and activated for 1 h at 120°C. Subsequently, samples with the methylated radiolabeled fatty acids together with unlabeled methylated carrier were applied to the plate. This process separated substrate (palmitic acid) from the unsaturated product palmitoleic acid. Plates were developed in developing solvent (hexane:ethyl ethe ⁇ acetic acid, 90:10:l,v:v:v) until the solvent front was about 2 cm from the top of the plate, then were dried and exposed to iodine overnight. Bands were visualized, scraped, and counted in scintillation counter.
  • SCDl inhibitors increase saturation of biliary phosphatidyl choline fatty acids.
  • Increasing saturation of biliary phospholipid fatty acids is known to increase enormously the cholesterol solubilizing capacity of bile.
  • SCDl inhibitors are very effective in treatment and prevention of cholesterol gallstones and other disorders of cholesterol crystallization in the biliary tree.
  • SCDl is the only enzyme in liver or the entire body performing delta 9-monodesaturation of 16:0 and 18:0 fatty acids.
  • the reduction in 16:1 and 18:1 observed in Examples 1-2 was expected to be due to SCDl inhibition, which was in fact demonstrated directly in liver microsomes in the present Example.
  • EXAMPLE 4 NON-FABAC SCDl INHIBITORS PREVENT GALLSTONE FORMATION
  • C57/BL6 mice were divided into 4 groups. All groups were fed the High Fat, Lithogenic, Diet described in Examples 1-2 above.
  • CLA Conjugated Linoleic Acid
  • RD regular diet
  • group 1 group 1
  • group 2 group 2
  • HFD high-fat diet
  • Six (6) mice in each group are sacrificed. No gallstones are found in the former group, while close to 100% of mice in the HFD group exhibit cholesterol gallstones.
  • Attie AD et al Relationship between stearoyl-CoA desaturase activity and plasma triglycerides in human and mouse hypertriglyceridemia. J Lipid Res. 2002 Nov;43(l l): 1899-907.

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Abstract

L'invention concerne le traitement d'affections liées à la cristallisation de cholestérol biliaire, particulièrement l'administration d'inhibiteurs de stéaroyl coenzyme A désaturase 1, sachant que l'inhibiteur de l'activité enzymatique de SCD1 n'est pas un conjugué d'acide biliaire et d'acide gras (FABAC).
PCT/IL2008/001467 2007-11-08 2008-11-06 Procédés et compositions pour le traitement de cristallisation de cholestérol biliaire et d'affections connexes Ceased WO2009060452A2 (fr)

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Publication number Priority date Publication date Assignee Title
WO2013056148A2 (fr) 2011-10-15 2013-04-18 Genentech, Inc. Procédés d'utilisation d'antagonistes de scd1
US20160175324A1 (en) * 2013-08-08 2016-06-23 Galderm Therapeutics Ltd. Anti-acne compositions comprising bile acid-fatty acid conjugates
US11571431B2 (en) 2013-12-04 2023-02-07 Galmed Research And Development Ltd Aramchol salts

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DE3430366A1 (de) * 1983-08-19 1985-03-07 Kao Corp., Tokio/Tokyo Mittel zur aufloesung von gallensteinen
JPS60172925A (ja) * 1984-02-17 1985-09-06 Kao Corp 胆石溶解剤
JPH0623099B2 (ja) * 1984-02-17 1994-03-30 花王株式会社 胆石溶解剤
GB9026648D0 (en) * 1990-12-07 1991-01-23 Efamol Holdings Nutrition
CN1110313C (zh) * 1998-03-18 2003-06-04 许秋菊 一种防治胆石病的药物
HUP0102048A3 (en) * 1998-04-16 2002-12-28 Eurovita As Novel synergistic compositions containing aromatic compounds and terpenoids present in alpinia galanga and their use
US20030064950A1 (en) * 2001-02-23 2003-04-03 Ntambi James M. Methods for reducing body fat and increasing lean body mass by reducing stearoyl-CoA desaturase 1 activity
WO2006023342A2 (fr) * 2004-08-20 2006-03-02 Tishcon Corp Combinaison synergique d'acide linoleique conjugue et de carnitine

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2013056148A2 (fr) 2011-10-15 2013-04-18 Genentech, Inc. Procédés d'utilisation d'antagonistes de scd1
US9358250B2 (en) 2011-10-15 2016-06-07 Genentech, Inc. Methods of using SCD1 antagonists
US20160175324A1 (en) * 2013-08-08 2016-06-23 Galderm Therapeutics Ltd. Anti-acne compositions comprising bile acid-fatty acid conjugates
US11571431B2 (en) 2013-12-04 2023-02-07 Galmed Research And Development Ltd Aramchol salts

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