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  • C07—ORGANIC CHEMISTRY
  • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
  • C07C69/00—Esters of carboxylic acids; Esters of carbonic or haloformic acids
  • C07C69/76—Esters of carboxylic acids having a carboxyl group bound to a carbon atom of a six-membered aromatic ring
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
  • C07C50/00—Quinones
  • C07C50/02—Quinones with monocyclic quinoid structure
  • C07C50/04—Benzoquinones, i.e. C6H4O2
• • 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
  • A61P1/18—Drugs for disorders of the alimentary tract or the digestive system for pancreatic disorders, e.g. pancreatic enzymes
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P3/00—Drugs for disorders of the metabolism
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P3/00—Drugs for disorders of the metabolism
  • A61P3/04—Anorexiants; Antiobesity agents
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P3/00—Drugs for disorders of the metabolism
  • A61P3/06—Antihyperlipidemics
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P3/00—Drugs for disorders of the metabolism
  • A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
  • A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
  • A61P31/04—Antibacterial agents
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P9/00—Drugs for disorders of the cardiovascular system
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P9/00—Drugs for disorders of the cardiovascular system
  • A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
  • C07C205/00—Compounds containing nitro groups bound to a carbon skeleton
  • C07C205/49—Compounds containing nitro groups bound to a carbon skeleton the carbon skeleton being further substituted by carboxyl groups
  • C07C205/57—Compounds containing nitro groups bound to a carbon skeleton the carbon skeleton being further substituted by carboxyl groups having nitro groups and carboxyl groups bound to carbon atoms of six-membered aromatic rings of the carbon skeleton
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
  • C07C69/00—Esters of carboxylic acids; Esters of carbonic or haloformic acids
  • C07C69/76—Esters of carboxylic acids having a carboxyl group bound to a carbon atom of a six-membered aromatic ring
  • C07C69/78—Benzoic acid esters
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
  • C07C69/00—Esters of carboxylic acids; Esters of carbonic or haloformic acids
  • C07C69/76—Esters of carboxylic acids having a carboxyl group bound to a carbon atom of a six-membered aromatic ring
  • C07C69/84—Esters of carboxylic acids having a carboxyl group bound to a carbon atom of a six-membered aromatic ring of monocyclic hydroxy carboxylic acids, the hydroxy groups and the carboxyl groups of which are bound to carbon atoms of a six-membered aromatic ring
  • C07C69/92—Esters of carboxylic acids having a carboxyl group bound to a carbon atom of a six-membered aromatic ring of monocyclic hydroxy carboxylic acids, the hydroxy groups and the carboxyl groups of which are bound to carbon atoms of a six-membered aromatic ring with etherified hydroxyl groups

Definitions

• the present invention relates to novel heterocyclic compounds of benzoquinones that affect the activity of enzymes of lipase gene family.
• the present invention relates to pharmaceutical compositions that contain the benzoquinone derived compounds, process for preparing the same and their pharmaceutically acceptable salts, derivatives, isomers, polymorphs, solvates thereof having a selective activity on diseases and conditions mediated by genes of lipase family.
• the first line of treatment for individuals suffering from such metabolic disorders involves adoption of a diet low in fat and regular exercise. Compliance with such regimen however can be poor and, as the disease progresses, treatment with therapeutic drugs becomes necessary.
• Hepatic lipase and lipoprotein lipase are multifunctional proteins which mediate the binding, uptake, catabolism, and remodeling of lipoproteins and phospholipids.
• Lipoprotein lipase and hepatic lipase function while bound to the luminal surface of endothelial cells in peripheral tissues and the liver respectively. Both enzymes participate in reverse cholesterol transport, which is the movement of cholesterol from peripheral tissues to the liver either for excretion from the body or for recycling.
• Genetic defects in both hepatic lipase and lipoprotein lipase are known to be the cause of familial disorders of lipoprotein metabolism. Defects in the metabolism of lipoproteins result in serious metabolic disorders, including hypercholesterolemia, hyperlipidemia, and atherosclerosis.
• the lipase gene family enzymes are involved in a wide array of metabolic pathways, ranging from lipid digestion, absorption, fatty acid uptake, lipoprotein transportation and also in inflammation (Wong Howard et al., 2002, The lipase gene family, Journal of Lipid Research, Vol. 43: 993-999).
• Pancreatic lipase is one of the key enzymes in lipid metabolism. It is synthesized by pancreatic acinar cells where it is secreted into the intestinal lumen and aids in the intestinal absorption of long chain triglyceride fatty acids (Verger, R. 1984, Pancreatic Lipases In Lipases. B. Borgstr ⁇ m and H.L. Brockman, editors. Elsevier, New York. 83- 150; Lowe, M. E. 1997, Molecular mechanisms of rat and human pancreatic triglyceride lipases. J. Nutr. 127: 549-557).
• Lipoprotein lipase is the major enzyme responsible for the distribution and utilization of triglycerides in the body. Lipoprotein lipase hydrolyzes triglycerides in both chylomicrons and VLDL. Hepatic lipase hydrolyzes triglycerides in IDL and HDL, and is responsible for lipoprotein remodeling. Hepatic lipase also functions as a phospholipase, and hydrolyzes phospholipids in HDL.
• Lipase members function in the metabolism of circulating lipoproteins. Hepatic lipase plays a role in the uptake of HDL cholesterol (Olivecrona, T., et al. 1993, Lipoprotein lipase and hepatic lipase. Curr. Opin. Lipidol. 4: 187-196). It is synthesized exclusively in the liver, where it is predominantly found (Hixenbaugh, E. A, et al., 1989, Hepatic lipase in the rat ovary. J. Biol. Chem. 264: 4222-4230).
• LPL lipoprotein lipase
• This lipase is bound to capillary endothelium, where it functions to supply the underlying tissue with fatty acids derived from the triglyceride-rich core of circulating chylomicrons and VLDL (Olivecrona, T., and G. Bengtsson-Olivecrona. 1993. Lipoprotein lipase and hepatic lipase. Curr. Opin. Lipidol. 4: 187-196). In the process, LPL transforms these lipoproteins into remnant and HDL particles.
• LPL produced by macrophages in the vascular wall may facilitate the development of atherosclerosis by promoting lipid accumulation within the lesion.
• LPL has been shown to be involved in the pathogenesis of atherosclerosis (Mead JR, et al. 1999, "Lipoprotein Lipase, a key role in atherosclerosis?" FEBS Lett., Nov 26, 462(1-2): 1-6).
• Transgenic animals expressing human lipoprotein lipase or hepatic lipase have decreased levels of plasma triglycerides and an increased level of high density lipoprotein (HDL) (Shimada, M., et al (1993) J. Biol. Chem. 268:17924-17929; Liu, M.-S., et al. (1994) J. Biol. Chem. 269:11417-11424).
• HDL high density lipoprotein
• a more recently discovered member of the lipase gene family is endothelial lipase.
• the drugs that inhibit or reduce the activity of lipases at various levels in the body form the front line of therapy for the treatment of diseases mediated by accumulation of fat at elevated levels.
• a lipase inhibitor that is marketed as anti-obesity drug include Orlistat (XENICAL ® ) is described in U.S. Patent No. 4598089.
• European Patent Application No. EP129748 relates to Orlistat and related compounds and their use in inhibiting pancreatic lipase and treating hyperlipidemia and obesity.
• Orlistat inhibits only intestinal lipases such as gastric, pancreatic and carboxylester lipases, particularly pancreatic lipase, in the gut lumen and blocks the digestion of dietary fat by preventing lipase from interacting with its lipid target.
• a plant benzoquinone embelin (2,5-dihydroxy-3-undecyl-l,4-benzoquinone) obtained from the dried fruit of Embelia ribes and known as an antifertility agent has also been reported to elevate activities of the lipogenic enzymes, malate dehydrogenase, glucose-6- phosphate dehydrogenase and hydroxymethylglutaryl-CoA reductase while essentially not affecting lipolytic enzyme activities.
• Embelin is also used as a teniacide, as having antitumor, anti-inflammatory and analgesic properties (Chitra et al.
• XIAP X-linked inhibitor of apoptosis
• the present invention provides compounds of Formula (I):
• R 1 and R 2 are each independently selected from the group consisting of hydrogen, C 3 -Ci 3 alkyl, Ci-C 20 haloalkyl, C 2 -Ci 3 alkenyl, C 2 -Ci 3 alkynyl, C 4 -C 6 cycloalkyl, C 4 -C 6 cycloalkenyl, Ci-Ci 3 alkoxyalkyl, Ci-C 5 alkylcycloalkyl, Ci-C 5 alkylcycloalkenyl, Ci-C 13 alkylamine, Ci-Ci 3 arylamine, C(O)Ci-C 6 alkyl, 0-C(O)Ci-C 6 alkyl, heterocycloalkyl, aryl, alkylaryl, C(O)aryl and O-C(O)aryl; wherein each of the foregoing groups may optionally bear 1 to 6 substituents independently selected from hydrogen, halo, nitro, amino, cyano, isocyano, thio
• the present invention also relates to the compounds of formula (I) and derivatives thereof including but not limited to polymorphs, isomers and prodrugs thereof, geometric , or optical isomers thereof, and pharmaceutically acceptable esters, ethers, carbamates of such compounds, all solvates and hydrates thereof and all salts thereof.
• present invention provides use of compounds of formula (I) in reducing or inhibiting metabolism, absorption, and accumulation of fat at various levels including fluid, cellular, and tissue levels in body by inhibiting or reducing the activity of enzymes belonging to lipase gene family.
• the invention provides compounds of formula (II):
• Ri and R 2 are each independently selected from the group consisting Of C 3 -Ci 3 alkyl, Ci- C 20 haloalkyl, C 2 -Ci 3 alkenyl, C 2 -Ci 3 alkynyl, C 4 -C 6 cycloalkyl, C 4 -C 6 cycloalkenyl, Ci- Ci 3 alkoxyalkyl, Ci-C 5 alkylcycloalkyl, Ci-C 5 alkylcycloalkenyl, Cj-Ci 3 alkylamine, Ci- Ci 3 arylamine, C(O)Ci-C 6 alkyl, heterocycloalkyl, aryl, alkylaryl, and C(O)aryl; wherein each of the foregoing groups may optionally bear 1 to 6 substituents independently selected from hydrogen, halo, nitro, amino, cyano, isocyano, thio, Ci-C 6 alkyl, cycloalkyl, aryl, alkoxy, and
• the present invention also relates to the compounds of formula (II) and derivatives thereof including but not limited to polymorphs, isomers and prodrugs thereof, geometric or optical isomers thereof, and pharmaceutically acceptable esters, ethers, carbamates of such compounds, all solvates and hydrates thereof and all salts thereof.
• present invention provides use of compounds of formula (II) in reducing or inhibiting metabolism, absorption, and accumulation of fat at various levels including fluid, cellular, and tissue levels in body by inhibiting or reducing the activity of enzymes belonging to lipase gene family.
• the invention provides compounds of formula (III):
• Ri and R 2 are each independently selected from the group consisting of hydrogen, Ci-Ci 3 alkyl, C r C 20 haloalkyl, C 2 -Ci 3 alkenyl, C 2 -Ci 3 alkynyl, C 4 -C 6 cycloalkyl, C 4 -C 6 cycloalkenyl, Ci-Ci 3 alkoxyalkyl, Ci-Ci 3 alkylamine, Ci-Ci 3 arylamine, C 1 -C 5 alkylcycloalkyl, C1-C5 alkylcycloalkenyl, C(O)Ci-C 6 alkyl, heterocycloalkyl, aryl, alkylaryl, and C(O)aryl; wherein each of the foregoing groups may optionally bear 1 to 6 substituents independently selected from hydrogen, halo, nitro, amino, cyano, isocyano, thio, Ci-C 6 alkyl, cycloalkyl, aryl, al
• the present invention also relates to the compounds of formula (III) and derivatives thereof including but not limited to polymorphs, isomers and prodrugs thereof, geometric or optical isomers thereof, and pharmaceutically acceptable esters, ethers, carbamates of such compounds, all solvates and hydrates thereof and all salts thereof.
• present invention provides use of compounds of formula (III) in reducing or inhibiting metabolism, absorption, and accumulation of fat at various levels including fluid, cellular, and tissue levels in body by inhibiting or reducing the activity of enzymes belonging to lipase gene family.
• the present invention further provides the process for preparation of compounds of formulas (I), (II) and (III) and derivatives thereof.
• the present invention provides a pharmaceutical compositions comprising any of the compounds of this invention including their polymorph, prodrug, isomer or pharmaceutically acceptable ester, ethers, carbamate, and oximes useful in reducing or inhibiting activity of enzymes of lipase gene family participating in metabolism, absorption, and accumulation of lipids in body at various levels including body fluid, cellular and tissue level for treatment, amelioration or prevention of diseases mediated by lipase gene family enzyme including but not limited to overweight or obesity, hyperlipidemia, hypercholesterolemia, hypertriglyceridemia, pancreatitis, hyperglycemia, atherosclerosis, metabolic syndromes, other cardiovascular diseases, and other metabolic disorders.
• the present invention in further aspect also provides the use of compounds of formulas (I), (II) and (III) and derivatives thereof for skin, hair care or cosmetic preparation.
• the present invention in still further aspect provides the use of compounds of formulas (I), (II) and (III) and derivatives thereof to prevent or treat cellular and tissue damage caused by microbial pathogens secreting Upases.
• the present invention also relates to the pharmaceutical formulations comprising of any of compound of formulas (I), (II) and (III) and derivatives thereof by themselves or in conjunction with a suitable pharmaceutically acceptable excipient.
• Such formulations are useful in reducing or inhibiting activity of enzymes of lipase gene family participating in metabolism, absorption, and accumulation of lipids in body at various levels including body fluid, cellular and tissue level for treatment, amelioration or prevention of diseases mediated by lipase gene family enzymes such as overweight or obesity, hyperlipidemia, hypercholesterolemia, hypertriglyceridemia, pancreatitis, diabetes, atherosclerosis, other cardiovascular diseases, metabolic syndromes, and metabolic disorders.
• the present invention also provides the manner of manufacture of medicaments comprising of compounds of formulas (I), (II) and (III) and derivatives thereof in a therapeutically effective amount either alone or in combination with pharmaceutically acceptable adjuvant.
• the compounds of formulas (I), (II) and (III) and derivatives thereof may further be combined with other active ingredients.
• the present invention further relates to the method of treatment of diseases mediated by lipase gene family of enzymes by administering in a therapeutically effective amount any compound of formulas (I), (II) and (III) and derivatives thereof in human or animal subjects.
• the present invention provides novel methods and compositions for use in reducing or inhibiting activity of lipase gene family enzymes for treatment, amelioration or prevention of lipase gene family enzyme mediated diseases and conditions in an individual.
• lipase gene family enzymes include but are not limited to hepatic lipase; intestinal lipases including gastric lipase, pancreatic lipase, and carboxylester lipase; endothelial lipase; phospholipase and other related lipases.
• pharmaceutically acceptable refers to the substance including carrier, diluent, vehicle excipient, or composition being compatible chemically and/or toxicologically, with the other ingredients comprising a formulation that is not deleterious to the recipient thereof.
• alkyl by itself or as part of another substituent means, unless otherwise defined, a straight or branched chain monovalent hydrocarbon radical, such as for example, methyl, ethyl, n-propyl, isopropyl, n-butyl, tertiary butyl, sec-butyl, n-pentyl and n-hexyl.
• alkenyl employed alone or in combination with other terms means a straight chain or branched monovalent hydrocarbon group having the stated number ranges of carbon atoms, and groups such as vinyl, propenyl, crotonyl, isopentenyl, and various butenyl isomers.
• alkynyl employed alone or in combination with other terms means a straight chain or branched acyclic carbon chain which contains a carbon-to-carbon triple bond hydrocarbon group having the stated number ranges of carbon atoms, and groups.
• cycloalkyl means a cyclic either monocyclic or polycyclic alkyl radical having at least 3 carbon atoms and typically 3 to 7 carbon atoms. Examples are: cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl or cycloheptyl.
• alkoxy signifies a group of the formula alkyl-O- in which the term “alkyl” has the previously given significance, such as methoxy, ethoxy, n- propoxy, isopropoxy, n-butoxy, isobutoxy, 2° butoxy and 3° butoxy, or 2- methoxyethoxy.
• C 1 to C 5 alkylcycloalkyl means any of the C 1 to C 5 alkyl group is substituted on the cycloalkyl group and the composite group is attached to the nucleus at the alkyl terminus.
• C 1 to C 6 heterocycloalkyl means a heterocycloalkyl group having 2-6 carbon atoms, preferably 3-5 carbon atoms, and including at least one heteroatom selected from N, O and/or S, which may be attached via a heteroatom or a carbon atom.
• aryl means an aromatic hydrocarbon group having a single (e.g. phenyl) or a fused ring system (e.g. naphthalene, anthracene, phenanthrene, etc.).
• a typical aryl group is aromatic carbocylic ring having 6, 7, 8, 9 or 10 carbon atoms, such as phenyl, naphthyl, tetrahydronaphthyl or indenyl, which may optionally be substituted with one or more substituents selected from hydroxy, amino, halogen, nitro, cyano, C 1 to C 4 alkyl, C 2 to C 4 alkenyl, C 2 to C 4 alkynyl, C 1 to C 4 alkoxy, Ci to C 4 dialkylamino, the alkyl moieties having the same meaning as previously defined.
• the preferred aromatic hydrocarbon group is phenyl.
• C 3 to Cg heteroaryl means a substituted or unsubstituted aromatic group having 3, 4, 5, 6, 7, 8 or 9 carbon atoms, at least including one heteroatom selected from N, O and/or S, like imidazolyl, thiadiazolyl, pyridyl, (benzo)thienyl, (benzo)furyl, quinolyl, tetrahydroquinolyl, quinoxalyl or indolyl.
• the substituents on the heteroaryl group may be selected from the group of substituents listed for the aryl group.
• the heteroaryl group may be attached via a carbon atom or a heteroatom, if feasible.
• C 6 to C 1 O aryloxy means an aryl group containing 6, 7, 8, 9, or 10 carbon atoms as defined previously, attached to an oxygen atom.
• C 3 to Cg heteroaryloxy groups are analogs of the C 6 to C 10 aryloxy groups, at least including one heteroatom selected from N, O or S.
• halo means fluoro, chloro, bromo, or iodo.
• amino signifies a primary, secondary or tertiary amino group bonded via the nitrogen atom, with the secondary amino group carrying an alkyl or cycloalkyl substituent and the tertiary amino group carrying two similar or different alkyl or cycloalkyl substituents or the two nitrogen substitutents together forming a ring, such as, for example, — -NH 2 , methylamino, ethylamino, dimethylamino, diethylamino, methyl-ethylamino, pyrrolidin-1-yl or piperidino etc., preferably amino, dimethylamino and diethylamino and particularly preferred primary amino.
• cyano alone or in combination, signifies a — CN group.
• nitro alone or in combination, signifies a — NO 2 group.
• heterocyclic group refers to radicals or groups derived from monocyclic or polycyclic saturated or unsaturated, substituted or unsubstiruted heterocyclic nuclei having 5, 6, 7, 8, 9, 10, 11, 12, 13 or 14 ring atoms and containing 1, 2 to 3 hetero atoms selected from the group consisting of nitrogen, oxygen or sulfur.
• substituent is "non-interfering" substituents.
• non-interfering is meant that the group is suitable chemically and stability wise to occupy the designated position and perform the designated or intended role. Thus unsuitable groups are excluded from the definition of "non-interfering”.
• compounds of Formula (I) and derivatives thereof may be labeled with an isotope (e.g., 3 H, 14 C, 35 S 5 125 I, etc.).
• an isotope e.g., 3 H, 14 C, 35 S 5 125 I, etc.
• a “prodrug” refers to a compounds capable of being converted to compounds of the present invention by reactions of an enzyme, gastric juice, or the like, under physiological conditions in vivo, specifically compounds capable of being converted to compounds of the present invention upon enzymatic oxidation, reduction, hydrolysis, or the like, or a compounds capable of being converted to compounds of the present invention upon hydrolysis or the like by gastric juice or the like.
• a “polymorph” refers to a compound that occurs in two or more forms.
• terapéuticaally effective amount means an amount of a compound of the present invention that - treat or prevent the particular disease, condition, or disorder; or attenuates, ameliorates, or eliminates one or more symptoms of the particular disease, condition, or disorder; or prevents o delays the onset of one or more symptoms of the particular disease, condition, or disorder described herein.
• Ci -C n is used to signify each of C 1 , C 2 , C 3 , ... C n .
• C 1 -C 20 includes C 1 , C 2 , C 3 , C 4 , C 5 , C 6 , C 7 , C 8 , C 9 , C 1O , Cn, C 12 , C 13 , Ci 4 , Ci 5 , Ci 6 , Cn, Ci 8 , C 19 , and C 20 .
• Ci-Ci 3 includes each of C 1 , C 2 , C 3 , C 4 , C 5 , C 6 , C 7 , C 8 , C 9 , Ci 0 , Cn, Ci 2 , Ci 3 .
• C 2 - Cj 3 includes each of C 2 , C 3 , C 4 , C 5 , C 6 , C 7 , C 8 , C 9 , C 10 , Cn, Ci 2 , Ci 3 , and so forth.
• NMR data is in the form of delta ( ⁇ ) values for major diagnostic protons given in parts per million (ppm) relative to tetramethylsilane (TMS) as internal standard determined at 300 MHz or 400 MHz using the indicated solvent.
• TMS tetramethylsilane
• Chemical symbols have their usual meanings : the following abbreviations have also been used : v( volume), w (weight), B>p. 9 boiling point), Mp.
• pancreatic lipase pancreatic lipase
• lipoprotein lipase lipoprotein lipase
• hepatic lipase Goldberg, I. J., Le, N. -A., Ginsberg, H. N., Krauss, R. M., and Lindgren, F. T. (1988) J. Clin. Invest. 81,561-568
• pancreatic lipase is primarily responsible for the hydrolysis of dietary lipids. Variants of pancreatic lipase have been described, but their physiological role has not been determined (Giller, T., Buchwald, P., Blum-Kaelin, D., and Hunziker, W. (1992) J. Biol. Chem. 267,16509-16516).
• the lipase polypeptides encoded by these lipase genes are approximately 450 amino acids in length with leader signal peptides to facilitate secretion.
• the lipase proteins are comprised of two principal domains (Winkler, K., D'Arcy, A., and Hunziker, W. (1990) Nature 343, 771-774).
• the amino terminal domain contains the catalytic site while the carboxyl domain is believed to be responsible for substrate binding, cofactor association, and interaction with cell receptors (Wong, H., Davis, R. C, Nikazy, J., Seebart, K. E., and Schotz, M. C. (1991) Proc. Natl. Acad. Sci.
• GXSXG triacylglycerol lipase family share a number of conserved structural features.
• One such feature is the "GXSXG” motif, in which the central serine residue is one of the three residues comprising the "catalytic triad" (Winkler, K., D'Arcy, A., and Hunziker, W. (1990) Nature 343, 771-774; Faustinella, F., Smith, L. C, and Chan, L. (1992) Biochemistry 31,7219-7223).
• conserved aspartate and histidine residues make up the balance of the catalytic triad.
• lid region A short span of 19-23 amino acids (the "lid region") forms an amphipathic helix structure and covers the catalytic pocket of the enzyme (Winkler, K., D'Arcy, A., and Hunziker, W. (1990) Nature 343, 771-774). Comparisons between hepatic and lipoprotein lipase have demonstrated that differences in triacylglycerol lipase and phospholipase activities of the enzymes are in part mediated by this lid region (Dugi, K. A., Dichek H. L., and Santamarina-Fojo, S. (1995) J. Biol. Chem. 270, 25396-25401).
• Triacylglycerol lipases possess varying degrees of heparin binding activity. Lipoprotein lipase has the highest affinity for heparin, and this binding activity has been mapped to stretches of positively charged residues in the amino terminal domain (Ma, Y., Henderson, H. E., Liu, M.-S., Zhang, H., Forsythe, I. J., Clarke-Lewis, L, Hayden, M. R., and Brunzell, J. D. J. Lipid Res. 35, 2049-2059).
• the present invention relates to the compounds of formula (I)
• Ri and R 2 are each independently selected from the group consisting of hydrogen, C 3 -Ci 3 alkyl, Ci-C 20 haloalkyl, C 2 -Ci 3 alkenyl, C 2 -Ci 3 alkynyl, C 4 -C 6 cycloalkyl, C 4 -C 6 cycloalkenyl, Ci-Ci 3 alkoxyalkyl, Ci -C 5 alkylcycloalkyl, Ci-C 5 alkylcycloalkenyl, Ci-C 13 alkylamine, Ci-Ci 3 arylamine, C(O)Ci-C 6 alkyl, 0-C(O)Ci-C 6 alkyl, heterocycloalkyl, aryl, al
• the invention relates to compounds of formula (II):
• Ri and R 2 are each independently selected from the group consisting of C 3 -Ci 3 alkyl, Ci-C 20 haloalkyl, C 2 -Ci 3 alkenyl, C 2 -Ci 3 alkynyl, C 4 -C 6 cycloalkyl, C 4 -C 6 cycloalkenyl, Ci-Ci 3 alkoxyalkyl, C1-C5 alkylcycloalkyl, Ci-C 5 alkylcycloalkenyl, Ci-Ci 3 alkylamine, Ci-Ci 3 arylamine, C(O)Ci-C 6 alkyl, heterocycloalkyl, aryl, alkylaryl, and C(O)aryl; wherein each of
• the invention relates to compounds of formula (III):
• R 1 and R 2 are each independently selected from the group consisting of hydrogen, Cj-Ci 3 alkyl, Q-C 2 0 haloalkyl, C 2 -Ci 3 alkenyl, C 2 -Cj 3 alkynyl, C 4 -C 6 cycloalkyl, C 4 -C 6 cycloalkenyl, Ci-Ci 3 alkoxyalkyl, Ci-Ci 3 alkylamine, Ci-Ci 3 arylamine, Ci-C 5 alkylcycloalkyl, Ci-C 5 alkylcycloalkenyl, C(O)Ci-C 6 alkyl, heterocycloalkyl, aryl, alkylaryl, and C(O)aryl; where
• compounds of Formula (III) have Ri and R 2 independently selected from C(O)aryl, C(O)alkylaryl, C(O)haloaryl, C(O)nitroaryll, or C(O)alkoxyaryl.
• Ri and R 2 are independently selected firom methyphenylcarbonyl, ethylphenylcarbonyl, propylphenylcarbonyl, butylphenylcarbonyl, chlorophenylcarbonyl, bromopheynylcabonyl, iodophenylcarbonyl, fluorophenylcarbonyl, nitrophenylcarbonyl, methoxyphenylcarbonyl, or ethoxyphenylcarbonyl.
• Ri and R 2 are independently selected from 2-methyphenylcarbonyl, 3-methyphenylcarbonyl, A- methyphenylcarbonyl, 4-ter-butylphenylcarbonyl, 2-chlorophenylcarbonyl, 3- chlorophenylcarbonyl, 4-chlorophenylcarbonyl, 2-bromopheynylcabonyl, 3- bromopheynylcabonyl, 4-bromopheynylcabonyl, 2-iodophenylcarbonyl, 3- iodophenylcarbonyl, 4-iodophenylcarbonyl, 2-fluorophenylcarbonyl, 3- fluorophenylcarbonyl, 4-fluorophenylcarbonyl, 2-nitrophenylcarbonyl, 3- nitrophenylcarbonyl, 4-nitrophenylcarbonyl, 2-methoxyphenylcarbonyl, 3- methoxyphenyl
• the present invention also encompasses prodrugs of compounds of the present invention.
• prodrug includes a compound that is transformed in vivo to yield a compound of Formulas (I), (II) or (III). Information about the use of prodrugs may be found in "Pro-drugs as Novel Delivery Systems," Vol. 14 of the A.C.S. Symposium Series, by T. Higuchi and W. Stella, and in Bioreversible Carriers in Drug Design, ed. Edward B. Roche, American Pharmaceutical Association and Pergamon Press, 1987.
• the compounds of the present invention may contain asymmetric or chiral centers, and therefore may exist in different stereoisomeric forms. All suitable optical isomers and stereoisomeric forms of the compounds of the present invention as well as mixtures thereof, including racemic mixtures, form part of the present invention.
• the present invention embraces all geometric and positional isomers. For example, if a compound of the present invention incorporates a double bond or a fused ring, the cis- and trans-forms, as well as mixtures, are embraced within the scope of the invention. With respect to such compounds, the present invention includes the use of a racemate, a single enantiomeric form, a single diastereomeric form, or mixtures thereof, as suitable.
• Such compounds may also exist as tautomers. Accordingly, the present invention relates to the use of all such suitable tautomers and mixtures thereof. Diastereomeric mixtures can be separated into their individual diastereoisomers on the basis of their physical chemical differences by methods well known to those skilled in the art, such as by chromatography and/or fractional crystallization.
• Enantiomers can be separated by converting the enantiomeric mixture into a diastereomeric mixture by reaction with an appropriate optically active compound (e.g., chiral auxiliary such as a chiral alcohol or Mosher's acid chloride), separating the diastereoisomers and converting (e.g., hydrolyzing) the individual diastereoisomers to the corresponding pure enantiomers or by resolution of the racemic form by recrystallization techniques, by synthesis from optically-active starting materials, by chiral synthesis, or by chromatographic separation using a chiral stationary phase. Also, some of the compounds of the present invention may be atropisomers (e.g., substituted biaryls) and are considered as part of this invention. Enantiomers can also be separated by use of a chiral HPLC column.
• an appropriate optically active compound e.g., chiral auxiliary such as a chiral alcohol or Mosher's acid chloride
• converting e.
• some compounds of the present invention may exhibit polymorphism.
• the scope of the present invention includes all polymorphic forms of the compounds according to the invention, which forms the further aspect of the invention. It is to be understood that the present invention encompasses any and all racemic, optically-active, polymorphic and stereoisomeric forms, or mixtures thereof, which form or forms possess properties useful in the treatment of the conditions indicated herein.
• the present invention also include isotopically-labeled compounds of the present invention which are identical to those recited herein, but for the fact that one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature.
• isotopes that can be incorporated into compounds of the invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, iodine, and chlorine, such as 2 H, 3 H 5 11 C, 13 C, 14 C, 13 N, 15 N, 15 0, 17 0, 18 0, 31 P, 32 P, 35 S, 18 F, 123 1, 125 I and 36 Cl, respectively.
• Certain isotopically-labeled compounds of the present invention are useful in compound and/or substrate tissue distribution assays.
• Tritiated (i.e.,. 3 H) and carbon-14 (i.e., 14 C) isotopes are particularly preferred for their ease of preparation and delectability.
• substitution with heavier isotopes such as deuterium (i.e., 2 H) may afford certain therapeutic advantages resulting from greater metabolic stability (e.g., increased in vivo half-life or reduced dosage requirements) and hence may be preferred in some circumstances.
• Positron emitting isotopes such as 15 O, 13 N, 11 C, and 18 F are useful for positron emission tomography (PET) studies to examine substrate receptor occupancy.
• Isotopically labeled compounds of the present invention can generally be prepared by procedures analogous to those disclosed in the Examples herein below, by substituting an isotopically labeled reagent for a non-isotopically labeled reagent.
• Scheme I depicts a general protocol for preparing compound of Formulas (I), (II) or (III) starting from 2,5-dihydroxy-3-undecyl-l,4-benzoquinone or its oxime, or substituted oxime, or suitable salt or analogs thereof.
• the starting material - 2,5-dihydroxy-3- undecyl-l,4-benzoquinone - is reacted with alkyl chloride or acyl chloride, or aryl chloride or aroyl chloride, or substituted aryl chloride or substituted aroyl chloride in a suitable inert halogenated solvent (e.g. dichloromethane) in presence of a suitable aromatic base (e.g.
• a suitable inert halogenated solvent e.g. dichloromethane
• a suitable aromatic base e.g.
• the compounds of the present invention as disclosed above are useful for reducing or inhibiting activity of lipase gene family enzymes for treatment, amelioration or prevention of lipase gene family enzyme mediated diseases.
• the compounds are useful in reducing or inhibiting metabolism, absorption, and accumulation of fat at various levels including body fluid, cellular, and tissue levels in body by inhibiting or reducing the activity of enzymes belonging to lipase gene family.
• the compounds of the present inventions and derivatives thereof including compositions thereof are useful in reducing or inhibiting activity of enzymes of lipase gene family participating in metabolism, absorption, and accumulation of lipids in body at various levels including body fluid, cellular and tissue level for treatment, amelioration or prevention of diseases mediated by lipase gene family enzyme including but not limited to overweight or obesity, hyperlipidemia, hypercholesterolemia, hypertriglyceridemia, pancreatitis, hyperglycemia, atherosclerosis, metabolic syndromes, other cardiovascular diseases, and other metabolic disorders.
• the compounds of the present invention and derivatives thereof including compositions thereof are useful in prevent or treat cellular and tissue damage caused by microbial pathogens secreting lipases.
• the compounds of the present invention and derivatives thereof including compositions thereof are also useful for skin, hair care or cosmetic preparation.
• the present. invention provides method for treating or preventing cellular and tissue damage caused by microbial pathogens secreting lipases by inhibiting or reducing the activity of enzymes belonging to lipase gene family in mammal including a human being which comprises administering to said mammal an effective treating amount of a compound of Formulas (I), (II) or (III) or derivatives thereof.
• a pharmaceutical composition comprising a therapeutically effective amount of a compound of Formulas (I), (II) or (III) or a derivative thereof and a pharmaceutically acceptable inert adjuvant, diluent or carrier.
• a pharmaceutical composition may comprise of at least one additional pharmaceutically active agent. Additional active pharmaceutical agent may be selected from chemically synthesized compounds or those derived from natural origin having desired pharmacological activity.
• a compound of Formulas (I), (II) or (III) or a derivative thereof can be administered in any conventional oral, buccal, nasal, by inhalation spray in unit dosage form, parenteral, (for example, intravenous, intramuscular, subcutaneous intrastemal or by infusion techniques), topical (for example, powder, ointment or drop), transdermal, intracisternal, intravaginal, intraperitoneal, intravesical, or rectal,.
• the compound of the present invention and at least one other pharmaceutically active agent may be administered either separately or in the pharmaceutical composition comprising both. It is generally preferred that such administration be oral. However, if the subject being treated is unable to swallow, or oral administration is otherwise impaired or undesirable, parenteral or transdermal administration may be appropriate.
• a compound of Formulas (I), (II) or (III) or a derivative thereof can be administered in the form of any modified release, controlled release or timed release formulations, (see, e.g., Langer, Science 249:1527-1533 (1990)).
• a pump can be used (Langer, Science 249:1527-1533 (1990); Sefton, CRC Crit. Ref. Biomed. Eng. 14:201 (1987); Buchwald et al, Surgery 88:507 (1980); and Saudek et al, N. Engl. J. Med. 321:57 r 4 (1989)).
• polymeric materials can be used (see Medical Applications of Controlled Release (Langer and Wise eds., 1974); Controlled Drug Bioavailability, Drug Product Design and Performance (Smolen and Ball eds., 1984); Ranger and Peppas, J. Macromol. Sci. Rev. Macromol. Chem. 23:61 (1983); Levy et al., Science 228:190 (1985); During et al., Ann. Neurol. 25:351 (1989); and Howard et al., J. Neurosurg. 71:105 (1989)).
• the dose of a compound of Formulas (I), (II) or (III) or derivatives thereof to be administered to a mammal including human or animal for the purposes as mentioned above is not specifically limited. Rather it is widely variable and subject to the pathologies, conditions, symptoms, or age of the subject and judgment of the attending physician or veterinarian.
• the general range of effective administration rates of the compounds of the present invention is from about 0.001 mg/kg body weight to about 100 mg/kg body weight of the subject per day.
• a preferred range of effective administration rates of the compounds of this invention is from about 0.01 mg/kg body weight to about 50 mg/kg body weight of the subject per day.
• Amounts are selected based on various factors, including the milieu to which the composition is administered, the site of the cells to be treated, the age, health, gender, and weight of a patient or animal to be treated, etc.
• Useful amounts include, 1, 5, 15, 20, 25, 30, 40, 60, 150, 200 milligrams, 1 gm, 2 gm, 3 gm, and ranges between 10 milligrams- 100 grams, 50 milligrams - 5 grams, 100 milligrams-10 grams, 250 milligrams-2.5 grams, 500 milligrams- 1.25 grams, etc., per dosage.
• the amount of compound of this invention will depend on such factors as the solubility of the compound, prodrug, isomer or pharmaceutically acceptable salt of this invention, the formulation used and the route of administration (e.g., orally, transdermally, parenterally or topically).
• Dosages of the compounds of the present invention can be administered to humans by any suitable route, with oral administration being preferable.
• Individual oral dosage form for example, tablets or capsules should generally contain from about 0.1 mg to about 100 mg of compound of this invention, in a suitable pharmaceutically acceptable vehicle, diluent or carrier.
• Dosages for intravenous administration are generally within the range of from about 0.1 mg to about 10 mg per single dose as required.
• the dosage is generally formulated as from about a 0.1% to about a 1% (w/v) solution.
• the physician will determine the actual dosage, which will be most suitable for an individual patient, and it will vary with, e.g., age, weight and response of the particular patient.
• the above dosages are exemplary of the average case but there can, of course, be individual instances where higher or lower dosage ranges are possible, such dosages of compounds of this invention, are within the scope of the present invention.
• the pharmaceutical formulation comprising a compound of Formulas (I), (II) or (III) or the derivatives thereof may be formulated in a conventional manner known to those skilled at the art using one or more pharmaceutically acceptable diluent, carrier, or vehicle.
• tablets can be prepared by methods known in pharmaceutical science by direct compression, by wet granulation, or by dry granulation.
• Their formulations usually incorporate diluents, binders, lubricants and disintegrators as well as a compound of this invention.
• Common diluents include, for example, various types of starch, lactose, mannitol, kaolin, calcium phosphate or sulfate, inorganic salts such as sodium chloride and powdered sugar. Powdered cellulose derivatives may also be used.
• Common tablet binders include substances such as starch, gelatin and sugars such as lactose, fructose, glucose and the like. Natural and synthetic gums are also convenient, including acacia, alginates, methylcellulose, polyvinylpyrrolidine and the like. Polyethylene glycol, ethylcellulose and waxes can also serve as binders.
• a lubricant is generally necessary in a tablet formulation to prevent the tablet and punches from sticking in the die.
• the lubricant is chosen from such slippery solids as talc, magnesium and calcium stearate, stearic acid and hydrogenated vegetable oils.
• Tablet disintegrators include substances, which swell when wetted to break up the tablet and release a compound, prodrug, isomer or pharmaceutically acceptable salt of this invention. They include starches, clays, celluloses, algins and gums.
• corn and potato starches methylcellulose, agar, bentonite, wood cellulose, powdered natural sponge, cation-exchange resins, alginic acid, guar gum, citrus pulp and carboxymethylcellulose, for example, may be used as well as sodium lauryl sulfate. Tablets are often coated with sugar as a flavor and sealant, or with film-forming protecting agents to modify the dissolution properties of the tablet.
• the compounds of the invention may also be formulated as chewable tablets, by using large amounts of pleasant-tasting substances such as mannitol in the formulation, as is now well- established in the art.
• a slowly soluble pellet of a compound of this invention may be prepared and incorporated in a tablet or capsule.
• the technique may be improved by making pellets of several different dissolution rates with subsequent filling the mixture of these pellets in capsules.
• Tablets or capsules may be coated with a film, which resists dissolution for a predictable period of time.
• Capsules can be prepared by mixing a compound of the invention with a suitable diluent and filling the proper amount of the mixture in capsules.
• suitable diluents include inert powdered substances such as starch of many different kinds, powdered cellulose, especially crystalline and macrocrystalline cellulose, sugars such as fructose, mannitol and sucrose, grain flours and similar edible powders.
• Liquid preparations for oral administration may take the form of, for example, solutions, syrups or suspensions, or they may be presented as a dry product for constitution with water or other suitable vehicle before use.
• Such liquid preparations may be prepared by conventional means with pharmaceutically acceptable additives such as suspending/viscosity enhancing agents (e.g. sorbitol syrup, methyl cellulose or hydrogenated edible fats); emulsifying agents (e.g. lecithin or acacia); non-aqueous vehicles (e.g. almond oil, oily esters or ethyl alcohol, medium chain triglycerides); and preservatives (e.g. methyl or propyl p-hydroxybenzoates or sorbic acid).
• suspending/viscosity enhancing agents e.g. sorbitol syrup, methyl cellulose or hydrogenated edible fats
• emulsifying agents e.g. lecithin or acacia
• non-aqueous vehicles e.g. almond
• the compounds of the invention may be formulated in the form of an injection, including using conventional catheterization techniques or infusion.
• Formulations for injection may be presented in unit dosage form e.g. in ampoules or in multi-dose containers, with an added preservative.
• the compositions may take such forms as suspensions, solutions or emulsions in oily or aqueous vehicles, and may contain formulating agents such as suspending, stabilizing and/or dispersing agents.
• the injectable solutions or suspensions may be formulated according to known art, using suitable non-toxic, parenterally-acceptable diluents or solvents, such as mannitol, 1,3- butanediol, water, Ringer's solution or isotonic sodium chloride solution, or suitable dispersing or wetting and suspending agents, such as sterile, bland, fixed oils, including synthetic mono- or diglycerides, and fatty acids, including oleic acid and surfactants such as, for example, hydroxypropyl cellulose, also the pH of the solution being suitably adjusted and buffered, where necessary.
• suitable non-toxic, parenterally-acceptable diluents or solvents such as mannitol, 1,3- butanediol, water, Ringer's solution or isotonic sodium chloride solution, or suitable dispersing or wetting and suspending agents, such as sterile, bland, fixed oils, including synthetic mono- or diglycerides, and fatty acids, including
• the active ingredient may be in powder form for reconstitution with a suitable vehicle, e.g. sterile pyrogen-free water, before use.
• a suitable vehicle e.g. sterile pyrogen-free water
• the parenteral preparations may also be made long-acting by dissolving or suspending a compound, prodrug, isomer or pharmaceutically acceptable salt of this invention, as the case may be, in oily or emulsified vehicles which allow it to disperse only slowly in the serum.
• the compounds of the present invention are conveniently delivered in the form of a solution or suspension from a pump spray container that is squeezed or pumped by the patient or as an aerosol spray presentation from a pressurized container or a nebulizer, with the use of a suitable propellant, e.g. dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas.
• a suitable propellant e.g. dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas.
• the dosage unit may be determined by providing a valve to deliver a metered amount.
• the pressurized container or nebulizer may contain a solution or suspension of the active compound.
• compositions When administered by nasal aerosol or inhalation, these compositions are prepared according to techniques well-known in the art of pharmaceutical formulation and may be prepared as solutions in saline, employing benzyl alcohol or other suitable preservatives, absorption promoters to enhance bioavailability, fluorocarbons, and/or other solubilizing or dispersing agents known in the art.
• Capsules and cartridges (made, for example, from gelatin) for use in an inhaler or insufflator may be formulated containing a powder mix of a compound of the invention and a suitable powder for inhalation base such as lactose or starch.
• the compounds of this invention may also be administered topically and this may be done by way of, e.g., creams, jellies, salves, lotions, gels, pastes, ointments, and the like, in accordance with standard pharmaceutical practice.
• the compounds of the present invention may also be administered transdermally (e.g., through the use of a patch).
• any suitable formulation for transdermal application comprising a compound of the present invention may be employed and such formulations would generally also contain a suitable transdermal carrier, e.g., an absorbable pharmacologically acceptable solvent to promote and assist passage of the compounds through the subject's skin.
• suitable transdermal devices may comprise the form of a bandage having a backing member and a reservoir containing the subject compound.
• bandage-type transdermal devices may further include suitable carriers, rate-controlling barriers, and means for securing the transdermal device to the subject's skin.
• any suitable base can be used.
• Cocoa butter is a traditional suppository base, which may be modified by the addition of waxes to raise its melting point.
• Water-miscible suppository bases comprising, particularly; polyethylene glycols of various molecular weights are in wide use.
• a compound of this invention may be incorporated in food, or beverages.
• the compounds of this invention may also be administered to a mammal other than a human.
• the method of administration and the dosage to be administered to such a mammal will depend, for example, on the animal species and the disease or disorder being treated.
• the compounds of this invention may be administered to animals in any suitable manner, e.g., orally, parenterally or transdermally, in any suitable form such as, for example, a capsule, bolus, tablet, pellet, e.g., prepared by admixing a compound, prodrug, isomer or pharmaceutically acceptable salt of this invention with a suitable diluent such as carbowax or carnuba wax together with a lubricant, liquid drench or paste, e.g., prepared by dispersing a compound of this invention in a pharmaceutically acceptable oil such as peanut oil, sesame oil or corn oil.
• a suitable diluent such as carbowax or carnuba wax together with a lubricant, liquid drench
• the compounds, prodrugs, isomers or pharmaceutically acceptable salts of this invention may also be administered to animals as an implant. Such formulations are prepared in a conventional manner in accordance with standard veterinary practice. As an alternative, the compounds of this invention may be administered with the water supply, e.g., in the form of a liquid or water-soluble concentrate. In addition, the compounds of this invention, e.g., within the pharmaceutical compositions of the invention, may be administered in the animal feedstuff, e.g., a concentrated feed additive or premix may be prepared for mixing with the normal animal feed, commonly along with a suitable carrier therefore.
• the animal feedstuff e.g., a concentrated feed additive or premix may be prepared for mixing with the normal animal feed, commonly along with a suitable carrier therefore.
• the carrier facilitates uniform distribution of the compound, prodrug, isomer or pharmaceutically acceptable salt of this invention in the, e.g., finished feed with which the premix is blended.
• Suitable carriers include, but are not limited to, liquids, e.g., water, oils such as soybean, corn, cottonseed, or volatile organic solvents, and solids, e.g., a small portion of the feed or various suitable meals including alfalfa, soybean, cottonseed oil, linseed oil, corncob, corn, molasses, urea and bone, and mineral mixes.
• the biological assays carried out using compounds of Formulas (I), (II) and (III) demonstrate potent inhibition of lipases.
• Powdered berries of Embelia ribes were extracted successively with petroleum ether, chloroform, ethyl acetate, methanol and water.
• the chloroform extract was subjected to repeated crystallization using petroleum ether as the crystallizing solvent.
• the crystallization step was repeated to increase the yield.
• the mother liquor was fractionated by silica gel column chromatography (100-200 mesh) using petroleum ether-chloroform as the eluting solvents (gradient elution) to further improve the yield. All fractions were monitored on Silica gel TLC plates (Silica gel 60 F 254 , Merck) with n-propanol:n- Butanoldiquor ammonia (6:1:3) as the TLC solvent system.
• test samples Structures #1 - #19 were dissolved in ImL DMSO to obtain a 10 mM stock solution, and were vortexed to dissolution and stored at 4°C. The various concentration of working samples were prepared in 0.2 M phosphate buffer, pH-8.0. This sample was used as test sample for all further assays.
• Lipase assay Lipase assay was performed by method described by Winkler and Stuckmann., 1979, with modification. (Winkler, U.K. & Stuckmann, M. Glycogen, hyaluronate, and some other polysaccharides greatly enhance the formation of exolipase by Serratia marcescens. J. Bacteriol.
• Assay was designed, using a 96-well format.
• the substrate used in this assay was p-nitro phenol palmitate (Sigma, Cat No- N-2752).
• 4.5 mg of p-nitro phenol palmitate was dissolved in 200 ⁇ l of N, N- dimethyl formamide (Sigma, Cat No, D-4551) and volume made up to 10 ml with 0.1 M pH 8.0 phosphate buffer.
• Pancreatic Lipase (Sigma, Cat No. L-3126) sample was prepared by dissolving the enzyme in 0. IM phosphate buffer at a concentration of 5 mg/ml.
• the reaction mixture consisted of 150 ⁇ l substrate solution; 40 ⁇ l phosphate buffer (pH 8.0, 0.2 M) and 10 ⁇ l lipase solution.
• the reaction mixture was incubated at 37 0 C and optical density was measured at 405 run after incubation. Enzyme activity was presented in the form of international unit (IU).
• Lipase activity One enzyme unit of lipase is defined as that quantity releasing lnm of free phenol from the substrate (p-nitro phenol palmitate) per min per ml under the standard assay condition (Winkler and Stuckmann, 1979; Yadav RP, Saxena RK, Gupta R, Davidson WS., Purification and characterization of a region-specific lipase from Aspergillus terreus. Biotechnol. Appl. Biochem. (1998) 28, (243-249)). It is derived from standard graph of p-nitro phenol (Sigma, 104-8).
• Lipase inhibition assay Enzyme inhibition assay was performed in a dose dependent manner . The concentration of the synthetic analogs checked were lOO ⁇ M & 200 ⁇ M. The assay was similar to assay described above except 40 ⁇ l of test sample was used instead of phosphate buffer in control. Optical density was measured at 0 hr and following incubation at 37 0 C.
• Enzyme inhibition was presented in the term of relative activity and percentage inhibition simply on the basis of change in international unit (IU).
• Table 1 Effect of compounds of structures #1 - #19 on pancreatic lipase inhibition.
• test samples Synthetic analogues were dissolved in 1ml DMSO to obtain a 10 mM stock solution. The samples were vortexed to dissolution and stored at 4 0 C. The various concentration of working samples were prepared in O.2 M phosphate buffer, pH- 8.0. This sample was used as test sample for all further assays.
• Lipase inhibition assay Enzyme inhibition assay was performed in a dose dependent manner . The concentration of the synthetic analogs checked were 6.25 ⁇ M-200 ⁇ M. The assay was similar to assay described above except 40 ⁇ l of test sample was used instead of phosphate buffer in control. Optical density was measured at 0 hr and following incubation at 37 0 C. The enzyme activity was measured in terms of international unit (IU).
• IU international unit
• Enzyme inhibition was presented in the term of % inhibition simply on the basis of change in international unit (IU).
• IC 5O calculation IC 50 of each analogue was calculated manually from dose dependent graph (6.25 ⁇ M-200 ⁇ M) of each analogue at the concentration, where the % inhibition of lipase was measured as 50% in two near straight points (above & below 50% inhibition). The value of IC5 0 was derived from linear regression. The value derived is based on interpolated data.
• Table 2 IC 50 of compounds of Structure #1 - Structure #19 for pancreatic lipase inhibition.
• Example 22 Inhibition of Lipid absorption.
• Overnight fasting 4-week-old male Wistar rats (weight range: 150-200 grams) were used for the lipid absorption study.
• the rats were divided into two groups of 6 rats each and lOO ⁇ l of blood was drawn from orbital sinus for estimation of plasma lipid profile at 0 hour.
• 1 ml of fat rich liquid diet was administered PO (par orally) by gavages.
• experimental group was given orally 100 ⁇ g of test compound (2,5-di-O- aroyl-3-undecyl-l,4-benzoquinone derivative Structures #1 - #19) dissolved in 500 ⁇ l of vehicle; control group received same volume of vehicle.
• Mouse macrophage cell line (J774A.1) were plated in 6- well culture plates (1 x 10 5 cells per well) in Dulbecco's Minimum Essential Medium (DMEM) containing 10% Fetal bovine serum. 100 ⁇ g of oxidized low density lipoproteins (LDL) was added to each well. In each plate one of the compounds (Structure #1- #19) (10 ⁇ M) was added to wells in triplicate, while 3 wells were maintained as control. Experiment was terminated after 48 hours. Cells were stained with oil Red and counterstained with hematoxylin and observed under microscope. As compared to control, treatment with Structure #2 - #19 significantly inhibited uptake of LDL by macrophages.
• DMEM Dulbecco's Minimum Essential Medium

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