EP0665845A1 - Fo-2546 substance, its production and its use as inhibitor of acyl-coa cholesterol acyltransferase - Google Patents
Fo-2546 substance, its production and its use as inhibitor of acyl-coa cholesterol acyltransferaseInfo
- Publication number
- EP0665845A1 EP0665845A1 EP93923038A EP93923038A EP0665845A1 EP 0665845 A1 EP0665845 A1 EP 0665845A1 EP 93923038 A EP93923038 A EP 93923038A EP 93923038 A EP93923038 A EP 93923038A EP 0665845 A1 EP0665845 A1 EP 0665845A1
- Authority
- EP
- European Patent Office
- Prior art keywords
- substance
- formula
- strain
- hydroxyl
- hydrogen
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Withdrawn
Links
- 239000000126 substance Substances 0.000 title claims abstract description 140
- 238000004519 manufacturing process Methods 0.000 title claims description 5
- 102000001494 Sterol O-Acyltransferase Human genes 0.000 title abstract description 13
- 108010054082 Sterol O-acyltransferase Proteins 0.000 title abstract description 13
- 239000003112 inhibitor Substances 0.000 title description 2
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 claims abstract description 26
- 244000005700 microbiome Species 0.000 claims abstract description 19
- 235000012000 cholesterol Nutrition 0.000 claims abstract description 11
- 238000012258 culturing Methods 0.000 claims abstract description 7
- 201000010099 disease Diseases 0.000 claims abstract description 5
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims abstract description 5
- 238000009825 accumulation Methods 0.000 claims abstract description 4
- 238000000034 method Methods 0.000 claims description 10
- 239000001257 hydrogen Substances 0.000 claims description 8
- 229910052739 hydrogen Inorganic materials 0.000 claims description 8
- 125000002887 hydroxy group Chemical group [H]O* 0.000 claims description 8
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 claims description 4
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 claims description 4
- 150000001875 compounds Chemical class 0.000 claims description 4
- 125000004435 hydrogen atom Chemical group [H]* 0.000 claims description 4
- 239000001301 oxygen Substances 0.000 claims description 4
- 229910052760 oxygen Inorganic materials 0.000 claims description 4
- 239000008194 pharmaceutical composition Substances 0.000 claims description 4
- 230000002401 inhibitory effect Effects 0.000 abstract description 7
- 230000002265 prevention Effects 0.000 abstract description 3
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 12
- 239000002609 medium Substances 0.000 description 11
- 238000005160 1H NMR spectroscopy Methods 0.000 description 7
- 238000001460 carbon-13 nuclear magnetic resonance spectrum Methods 0.000 description 7
- 238000002329 infrared spectrum Methods 0.000 description 7
- 238000001228 spectrum Methods 0.000 description 7
- 238000002211 ultraviolet spectrum Methods 0.000 description 7
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 6
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 6
- UHOVQNZJYSORNB-UHFFFAOYSA-N Benzene Chemical compound C1=CC=CC=C1 UHOVQNZJYSORNB-UHFFFAOYSA-N 0.000 description 6
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 6
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 6
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- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 4
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- 208000035150 Hypercholesterolemia Diseases 0.000 description 3
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 description 3
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 3
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- 229910052757 nitrogen Inorganic materials 0.000 description 3
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- 229940123324 Acyltransferase inhibitor Drugs 0.000 description 2
- DKPFZGUDAPQIHT-UHFFFAOYSA-N Butyl acetate Natural products CCCCOC(C)=O DKPFZGUDAPQIHT-UHFFFAOYSA-N 0.000 description 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
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- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 2
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 2
- 125000002252 acyl group Chemical group 0.000 description 2
- 239000002404 acyltransferase inhibitor Substances 0.000 description 2
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 2
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- WORJEOGGNQDSOE-UHFFFAOYSA-N chloroform;methanol Chemical compound OC.ClC(Cl)Cl WORJEOGGNQDSOE-UHFFFAOYSA-N 0.000 description 2
- 150000001840 cholesterol esters Chemical class 0.000 description 2
- 230000008021 deposition Effects 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-M dihydrogenphosphate Chemical compound OP(O)([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-M 0.000 description 2
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- 239000000945 filler Substances 0.000 description 2
- 235000011187 glycerol Nutrition 0.000 description 2
- FUZZWVXGSFPDMH-UHFFFAOYSA-N hexanoic acid Chemical compound CCCCCC(O)=O FUZZWVXGSFPDMH-UHFFFAOYSA-N 0.000 description 2
- 229910017053 inorganic salt Inorganic materials 0.000 description 2
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- OCKPCBLVNKHBMX-UHFFFAOYSA-N n-butyl-benzene Natural products CCCCC1=CC=CC=C1 OCKPCBLVNKHBMX-UHFFFAOYSA-N 0.000 description 2
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- 239000012138 yeast extract Substances 0.000 description 2
- 229920001817 Agar Polymers 0.000 description 1
- GUBGYTABKSRVRQ-XLOQQCSPSA-N Alpha-Lactose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@H](O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-XLOQQCSPSA-N 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- VEXZGXHMUGYJMC-UHFFFAOYSA-M Chloride anion Chemical compound [Cl-] VEXZGXHMUGYJMC-UHFFFAOYSA-M 0.000 description 1
- 239000004375 Dextrin Substances 0.000 description 1
- 229920001353 Dextrin Polymers 0.000 description 1
- PLUBXMRUUVWRLT-UHFFFAOYSA-N Ethyl methanesulfonate Chemical compound CCOS(C)(=O)=O PLUBXMRUUVWRLT-UHFFFAOYSA-N 0.000 description 1
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- 108010010803 Gelatin Proteins 0.000 description 1
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- 108010013563 Lipoprotein Lipase Proteins 0.000 description 1
- 102100022119 Lipoprotein lipase Human genes 0.000 description 1
- VZUNGTLZRAYYDE-UHFFFAOYSA-N N-methyl-N'-nitro-N-nitrosoguanidine Chemical compound O=NN(C)C(=N)N[N+]([O-])=O VZUNGTLZRAYYDE-UHFFFAOYSA-N 0.000 description 1
- 235000019483 Peanut oil Nutrition 0.000 description 1
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- 108010009736 Protein Hydrolysates Proteins 0.000 description 1
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- DBMJMQXJHONAFJ-UHFFFAOYSA-M Sodium laurylsulphate Chemical compound [Na+].CCCCCCCCCCCCOS([O-])(=O)=O DBMJMQXJHONAFJ-UHFFFAOYSA-M 0.000 description 1
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- 230000004913 activation Effects 0.000 description 1
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- 150000004781 alginic acids Chemical class 0.000 description 1
- YNQLUTRBYVCPMQ-UHFFFAOYSA-N alpha-methyl toluene Natural products CCC1=CC=CC=C1 YNQLUTRBYVCPMQ-UHFFFAOYSA-N 0.000 description 1
- PNEYBMLMFCGWSK-UHFFFAOYSA-N aluminium oxide Inorganic materials [O-2].[O-2].[O-2].[Al+3].[Al+3] PNEYBMLMFCGWSK-UHFFFAOYSA-N 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 150000003863 ammonium salts Chemical class 0.000 description 1
- FROZIYRKKUFAOC-UHFFFAOYSA-N amobam Chemical compound N.N.SC(=S)NCCNC(S)=S FROZIYRKKUFAOC-UHFFFAOYSA-N 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- 239000002518 antifoaming agent Substances 0.000 description 1
- 239000007900 aqueous suspension Substances 0.000 description 1
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- 239000005018 casein Substances 0.000 description 1
- BECPQYXYKAMYBN-UHFFFAOYSA-N casein, tech. Chemical compound NCCCCC(C(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(CC(C)C)N=C(O)C(CCC(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(C(C)O)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(COP(O)(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(N)CC1=CC=CC=C1 BECPQYXYKAMYBN-UHFFFAOYSA-N 0.000 description 1
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- QIQXTHQIDYTFRH-UHFFFAOYSA-N octadecanoic acid Chemical compound CCCCCCCCCCCCCCCCCC(O)=O QIQXTHQIDYTFRH-UHFFFAOYSA-N 0.000 description 1
- 239000003921 oil Substances 0.000 description 1
- 235000019198 oils Nutrition 0.000 description 1
- XDUHQPOXLUAVEE-BPMMELMSSA-N oleoyl-CoA Chemical compound O[C@@H]1[C@H](OP(O)(O)=O)[C@@H](COP(O)(=O)OP(O)(=O)OCC(C)(C)[C@@H](O)C(=O)NCCC(=O)NCCSC(=O)CCCCCCC\C=C/CCCCCCCC)O[C@H]1N1C2=NC=NC(N)=C2N=C1 XDUHQPOXLUAVEE-BPMMELMSSA-N 0.000 description 1
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- 229940127557 pharmaceutical product Drugs 0.000 description 1
- 239000008055 phosphate buffer solution Substances 0.000 description 1
- 150000003013 phosphoric acid derivatives Chemical class 0.000 description 1
- 229920001223 polyethylene glycol Polymers 0.000 description 1
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- 235000013855 polyvinylpyrrolidone Nutrition 0.000 description 1
- XAEFZNCEHLXOMS-UHFFFAOYSA-M potassium benzoate Chemical compound [K+].[O-]C(=O)C1=CC=CC=C1 XAEFZNCEHLXOMS-UHFFFAOYSA-M 0.000 description 1
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- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 description 1
- 235000019333 sodium laurylsulphate Nutrition 0.000 description 1
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- QORWJWZARLRLPR-UHFFFAOYSA-H tricalcium bis(phosphate) Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O QORWJWZARLRLPR-UHFFFAOYSA-H 0.000 description 1
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Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D491/00—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00
- C07D491/12—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00 in which the condensed system contains three hetero rings
- C07D491/20—Spiro-condensed systems
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D491/00—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00
- C07D491/22—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00 in which the condensed system contains four or more hetero rings
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
- C12N1/145—Fungal isolates
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P17/00—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms
- C12P17/18—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms containing at least two hetero rings condensed among themselves or condensed with a common carbocyclic ring system, e.g. rifamycin
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/645—Fungi ; Processes using fungi
Definitions
- the present invention relates to a novel FO-2546 substance having an inhibitory activity for acylcoenzyme A cholesterol acyltransferase (hereinafter designated as acyl CoA cholesterol acyltransferase).
- hyperlipemia is known to be a factor for stimulating a progress of arteriosclerosis, and hence ischemic cardiopathy can be decreased by lowering blood cholesterol level.
- ischemic cardiopathy can be decreased by lowering blood cholesterol level.
- the crisis of yocardial infraction caused by hyperlipemia is observed at a high rate. Accordingly more effective and safe medicament for treatment of hyperlipemia, in particular hypercholesterolemia, is desirable.
- Cholesterol is converted to cholesterol ester by an action of acyltransfer from acyl CoA which is accumulated in the cells and in blood lipoprotein.
- An enzyme which catalyses acyltransfer reaction is acyl CoA cholesterol acyltransferase, and is specifically related to adsorption of cholesterol from the intestinal tract and a formation of the foam cells in the coronary artery.
- acyl CoA cholesterol acyltransferase inhibitor is estimated to be effective for treatment of the hypercholesterolemia.
- acyl CoA cholesterol acyltransferase inhibitor is expected to be useful for treatment of the diseases of adult people such as hyperlipemia or arteriosclerosis attributed therefrom.
- An object of the present invention is to provide FO-2546 substance of the formula
- R . is hydrogen or hydroxyl, and W is no bonding or oxygen, X is, together with Y, a group
- Another object of the present invention is to provide F0-2546A substance, F0-2546B substance, F0-2546C substance, FO- 2546D substance, F0-2546I substance, F0-2546J substance and FO- 2546K substance.
- Further object of the present invention is to provide a process for production of FO-2546 substance which comprises culturing the FO-2546 substance producing microorganism belonging to Coelomycetes in a nutrient medium, accumulating the FO-2546 substance therein, and isolating the FO-2546 substance therefrom.
- More further object of the present invention is to provide a microorganism strain belonging to Coelomycetes having an ability to produce FO-2546 substance.
- More further object of the present invention is to provide are pharmaceutical composition for treatment and prevention of a disease attributed of cholesterol, and also provide of a method of treating hypercholesterolemia in mammals.
- the microorganism having an ability to produce FO-2546 substance belongs to Coelomycetes.
- FO-2546 substance producing microorganism belongs to Coelomycetes.
- a microorganism strain FO-2546 belonging to Coelomycetes isolated by the inventors of the present invention is one of the most preferable strain used in the process of the present invention.
- Taxonomical properties of the strain FO-2546 are shown in the followings.
- Taxonomical properties 1. Observation on various media: (1) Cultural observations of the strain are illustrated in Table 1. Observations were taken macroscopically after cultivation at 25°C for 14 days. Table 1
- the present Strain FO-2546 is defined as the strain belonging to Coelomycetes, however the strain is not detailed identified in its taxonomical position. Therefore, the present strain is given a temporary name Strain FO-2546.
- the strain has been deposited in the National Institute of Bioscience and Human-Technology, Agency of Industrial Science and Technology of 1-3, Higashi 1-chome Tsukuba-shi Ibaraki-ken Japan with the deposition No. FERM BP-4406. The deposition was made on July 17th, 1992.
- a microorganism FO-2546 substance producing strain belonging to Coelomycetes is cultured in a nutrient medium.
- a culturing method generally used for fungi can be used on the present invention.
- a nutrient medium containing assimilable carbon source, assimilable nitrogen source and, if required together with inorganic salt can be used.
- assimilable carbon sources are glucose, sucrose, molasses, dextrin or cellulose etc. and combinations thereof.
- assimilable nitrogen sources are organic nitrogen source such as peptone, meat extract, yeast extract, dry yeast, soybean powder, corn steep liquor, cotton seed cake, casein, soybean protein hydrolysate, amino acid or urea, and inorganic nitrogen source such as ammonium salt, and combinations thereof.
- organic nitrogen source such as peptone, meat extract, yeast extract, dry yeast, soybean powder, corn steep liquor, cotton seed cake, casein, soybean protein hydrolysate, amino acid or urea
- inorganic nitrogen source such as ammonium salt, and combinations thereof.
- inorganic salt such as sodium salt, potassium salt, calcium salt, magnesium salt and phosphate salt, and heavy metallic salt can be added to the medium.
- trace nutrient element, growth stimulant or precusor can be added in the medium, if desired.
- Cultivation should be performed under aerobic condition, e. g., by shaking or agitation. Submerged aeration agitation cultivation ispreferable for industrial production. Cultivation can be preferably performed at neutral pH. Cultivation temperature is usually at 20- 37 ⁇ C. preferably at 24 - 30°C. FO-2546 substance is accumulated in the cultured mass by culturing for 2 - 3 days, and when maximum production of the FO-2546 substance is obtained, cultivation can be terminated.
- the FO- 2546 substance is accumulated in mycelia and in the cultured broth.
- the FO-2546 substance can preferably be isolated from supernatant and mycelia separated by centrifugation of cultured broth.
- the FO-2546 substance can be extracted from supernatant with water-immiscible organic solvent such as ethyl acetate, butyl acetate or benzene, and crude FO-2546 substance can be obtained by concentrating the extract in vacuo.
- the crude substance is subjected to a known purification process for 1ipophilie substance, for example, by column chromatography using a carrier such as silica gel or alumina.
- a carrier such as silica gel or alumina.
- Each component of the FO-2546 substance such as F0-2546A, -B, -C, -D, -I, -J and -K substance can be isolated.
- the FO-2546 substance can also be isolated from mycelial cells by extracting the mycelia with water- iscible organic solvent such as aqueous acetone or aqueous methanol.
- the extract is concentrated under reduced pressure and the concentrate is subjected to further extraction with water-immiscible organic solvent such as ethyl acetate, butyl acetate or benzene.
- the extract which is optionally combined with the extract obtained from cultured filtrate hereinabove explained, can be purified, or it can be purified and separated by the same procedure as of the above described to obtained each component of the FO-2546 substance.
- F0-2546A substance F0-2546B substance, F0-2546C substance, F0-2546D substance, F0-2546I substance, F0-2546J substance and F0-2546K substance are shown as follows.
- Radioactivity incorporated in the cholesterol ester fraction is analysed by RI scanner (Ambis Corp.) to assay an activity of acyl CoA cholesterol acyltransferase.
- the 50 % inhibition concentration on the enzyme is shown in Table 4.
- FO-2546 substance and the individual components thereof, of the present invention show
- the substances are useful in the treatment of high serum cholesterol in mammals, including humans.
- treatment is meant to include both the prevention and alleviation of high serum cholesterol.
- the substances may be administered to a subject in need of treatment by a variety of conventional routes of administration, including oral and parenteral. In general, these substances will be administered orally or parenterally at dosages between about 0.1 and about 10 mg/kg body weight of the subject to be treated per day, preferably from about 0.5 to 5 mg/kg. However, some variation in dosage will necessarily occur depending on the condition of the subject being treated and the activity of the compound being employed. The person responsible for administration will, in any event, determine the appropriate dose for the individual subject.
- a substance of the invention may be administered alone or in combination with pharmaceutically acceptable carriers, in either single or multiple doses.
- suitable pharmaceutically carriers include inert solid diluents or fillers, sterile aqueous solution and various organic solvents.
- the resulting pharmaceutical compositions are then readily administered in a variety of dosage forms such as tablets, powders, lozenges, syrups, injectable solutions and the like.
- These pharmaceutical compositions can, if desired, contain additional ingredients such as flavorings, binders, excipients and the like.
- tablets containing various excipients such as sodium citrate, calcium carbonate and calcium phosphate may be employed along with various disintegrants such as starch, alginic acid and certain complex silicates, together with binidng agents such as polyvinylpyrrolidone, sucrose, gelatin and acacia.
- binidng agents such as polyvinylpyrrolidone, sucrose, gelatin and acacia.
- lubricating agents such as magnesiu stearate, sodium lauryl sulfate and talc are often useful for tabletting purposes.
- Solid compositions of a similar type may also be employed as fillers in sort and hard filled gelatin capsules. Preferred materials for this include lactose or milk sugar and high molecular weight polyethylene glycols.
- the essential active ingredient therein may be combined with various sweetening or flavoring agents coloring matter or dyes and. if desired, emulsifying or suspending agents, together with diluents such as water, ethanol, propylene glycol, glycerin and combinations thereof.
- solutions of substance of the invention in seasa e or peanut oil, aqueous propylene glycol, or in sterile aqueous solution may be employed.
- aqueous solutions should be suitably buffered if necessary and the liquid diluent first rendered isotonic with sufficient saline or glucose.
- Such solutions are especially suitable for intravenous, intramuscular, subcutaneous and intraperitioneal administration.
- FIG. 1 UV spectrum of F0-2546A substance
- Figure 2 IR spectrum of F0-2546A substance
- Figure 3 'H-NMR spectrum of F0-2546A substance
- Figure 4 13 C-NMR spectrum of F0-2546A substance
- Figure 5 UV spectrum of F0-2546B substance
- Figure 6 IR spectrum of F0-2546B substance
- Figure 7 'H-NMR spectrum of F0-2546B substance
- Figure 8 13 C-NMR spectrum of F0-2546B substance.
- Figure 9 UV spectrum of F0-2546C substance
- Figure 10 IR spectrum of F0-2546C substance
- Figure 11 'H-NMR spectrum of 2546C substance.
- Figure 12 13 C-NMR spectrum of F0-2546C substance
- Figure 13 UV spectrum of F0-2546D substance.
- Figure 14 IR spectrum of F0-2546D substance
- Figure 15 'H-NMR spectrum of 2546D substance
- Figure 16 13 C-NMR spectrum of F0-2546D substance.
- Figure 17 UV spectrum of F0-2546I substance
- Figure 18 IR spectrum of F0-2546I substance
- Figure 19 'H-NMR spectrum of F0-2546I substance
- Figure 20 13 C-NMR spectrum of F0-2546I substance.
- Figure 21 UV spectrum of F0-2546J substance
- Figure 22 IR spectrum of F0-2546J substance
- Figure 23 'H-NMR spectrum of 2546J substance
- Figure 24 13 C-NMR spectrum of F0-2546J substance
- Figure 25 UV spectrum of F0-2546K substance
- Figure 26 IR spectrum of F0-2546K substance
- Figure 27 'H-NMR spectrum of 2546K substance
- Figure 28 13 C-NMR spectrum of F0-2546K substance.
- One loopful microorganism strain F0-2546 FERM BP-4406 grown on a nutrient agar medium was inoculated aseptically into the medium and shake cultured at 27°C for 48 hours to prepare seed culture liquid.
- a medium comprising soluble starch 3. 0 %, glycerol 1.0 1 soybean powder 2.01 dry yeast 0.31 potasium chloride 0.31 calcium carbonate 0.21 MgS0 4 • 7H 2 0 0.05 % and potasium dihydrogen phosphate 0.05 % (pH adjusted to 6.5) in 30 lit.
- jar- fer enter was sterilized and cooled.
- the seed culture 200 ml was transferred aseptically thereto and the inoculated medium was cultured aerobically at agitation 250 rp , aeration 10 lit./min. at 27°C for 72 hours.
- Cultured liquid 20 lit. including mycelia was extracted with ethyl acetate 18 lit. The extract was concentrated under reduced pressure to obtain crude substance 19g.
- the crude substance was charged on a column of silica gel (400g, Merk, Art 9385) and eluted with a mixture of chloroform- methanol (99 : 1). Active fractions containing FO-2546 substance comprising each 100 ml fraction were collected and concentrated in vacuo to obtain the crude substance 1.0 g.
- the crude substance was purified separately divided with 10 times by high performance liquid chromatography (Trirota V, Nihon Bunko Co.. column YMC-Pack A-343, ODS resin. Yama ura Kagaku Kenkyusho) with linear gradient elution solvent sysytem of 75 - 90 % aqueous acetonitrile for 20 min., detection UV 225 nm, flow rate 8 ml/min.
- F0-2546B substance 4.6 mg.
- F0-2546I substance 41 mg, F0-2546J substance 134 mg and F0-2546K substance 11 mg were obtained.
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Abstract
FO-2546 substance of formula (I) comprising FO-2546A substance, FO-2546B substance, FO-2546C substance and FO-2546D substance, FO-2546I substance, FO-2546J substance and FO-2546K substance, which has inhibitory activity for acylcoenzyme A cholesterol acyltransferase is produced by culturing microorganism strain FO-2546 belonging to Coelomycetes. The substance is useful for treatment and prevention of a disease attributed by accumulation of cholesterol.
Description
FO-2546 SUBSTANCE ITS PRODUCTION AND ITS USE AS INHIBITOR, OF ACYL-COA CHOLESTEROL ACYLTRANSFERASE
The present invention relates to a novel FO-2546 substance having an inhibitory activity for acylcoenzyme A cholesterol acyltransferase (hereinafter designated as acyl CoA cholesterol acyltransferase).
Heretofore some pharmaceutical products for treatment of hyperlipemia have been reported. Among them, a drug having an action on (1) cholesterol synthesis inhibition, (2) cholesterol absorption inhibition, (3) stimulation of cholesterol catabolism and (4) activation of lipoprotein lipase (suppression of lipoprotein synthesis), has been known.
In recent years, the symptoms attributable to accumulation of cholesterol in vivo such as hyperlipemia and arteriosclerosis in adult are becoming world-wide problems. Hyperlipemia is known to be a factor for stimulating a progress of arteriosclerosis, and hence ischemic cardiopathy can be decreased by lowering blood cholesterol level. The crisis of yocardial infraction caused by hyperlipemia is observed at a high rate. Accordingly more effective and safe medicament for treatment of hyperlipemia, in particular hypercholesterolemia, is desirable.
Cholesterol is converted to cholesterol ester by an action of acyltransfer from acyl CoA which is accumulated in the cells and in blood lipoprotein. An enzyme which catalyses acyltransfer reaction is acyl CoA cholesterol acyltransferase, and is specifically related to adsorption of cholesterol from the intestinal tract
and a formation of the foam cells in the coronary artery. Thererfore acyl CoA cholesterol acyltransferase inhibitor is estimated to be effective for treatment of the hypercholesterolemia. To provide acyl CoA cholesterol acyltransferase inhibitor is expected to be useful for treatment of the diseases of adult people such as hyperlipemia or arteriosclerosis attributed therefrom.
We have found according to continued studies on the metabolic products produced by microorganisms that substances having an inhibitory activity for acyl CoA cholesterol acyltr¬ ansferase are produced in a fermentation broth of microorganism Strain FO-2546 isolated from soil sample. Further we have isolated the substances having inhibitory activity for acyl CoA cholesterol acyltransferase which have physico-chemical properties described hereinbelow. These substances are novel compounds and were designated as F0-2546A substance, F0-2546B substance, F0-2546C substance, F0-2546D substance, F0-2546I substance, F0-2546J substance and F0-2546K substance. In the present invention, these substances were given a general name FO-2546 substance.
An object of the present invention is to provide FO-2546 substance of the formula
wherein R. is hydrogen or hydroxyl, and W is no bonding or oxygen, X is, together with Y, a group
0 in which R2 is -CH= C (CH3 ) 2 or -CH - C (CH3 ) and R3 is hydrogen or hydroxyl.
Another object of the present invention is to provide F0-2546A substance, F0-2546B substance, F0-2546C substance, FO- 2546D substance, F0-2546I substance, F0-2546J substance and FO- 2546K substance.
Further object of the present invention is to provide a process for production of FO-2546 substance which comprises culturing the FO-2546 substance producing microorganism belonging to Coelomycetes in a nutrient medium, accumulating the FO-2546 substance therein, and isolating the FO-2546 substance therefrom.
More further object of the present invention is to provide a microorganism strain belonging to Coelomycetes having an ability to produce FO-2546 substance.
More further object of the present invention is to provide are pharmaceutical composition for treatment and prevention of a disease attributed of cholesterol, and also provide of a method of
treating hypercholesterolemia in mammals.
The microorganism having an ability to produce FO-2546 substance (herein designated as FO-2546 substance producing microorganism) belongs to Coelomycetes. For example a microorganism strain FO-2546 belonging to Coelomycetes isolated by the inventors of the present invention is one of the most preferable strain used in the process of the present invention.
Taxonomical properties of the strain FO-2546 are shown in the followings.
Taxonomical properties: 1. Observation on various media: (1) Cultural observations of the strain are illustrated in Table 1. Observations were taken macroscopically after cultivation at 25°C for 14 days. Table 1
(To be continued Table 1)
2. Physiological properties:
(1) Growth temperature 8.5 - 33°C
(2) Optimum growth temperature 18.5 - 28°C
(3) Growth pH 2 - 10
(4) Optimum growth pH 3 - 6
(5) Nature aerobic
In comparison with the above properties of the Strain FO- 2546 based upon the morphological properties, various properties
on culture and physiological properties, and the taxonomical properties of known microorganism species, the present Strain FO-2546 is defined as the strain belonging to Coelomycetes, however the strain is not detailed identified in its taxonomical position. Therefore, the present strain is given a temporary name Strain FO-2546. The strain has been deposited in the National Institute of Bioscience and Human-Technology, Agency of Industrial Science and Technology of 1-3, Higashi 1-chome Tsukuba-shi Ibaraki-ken Japan with the deposition No. FERM BP-4406. The deposition was made on July 17th, 1992.
Although a description of the Strain FO-2546 was given hereinabove, taxonomical properties of the microorganism strain are in general very mutable and are not stable. Spontaneous mutation and artificial mutation with ultraviolet radiation or treatment with mutagen such as N-methyl-N-nitro-N-nitrosoguanidine or ethyl methanesulfonate or microorganisms can occur. Therefore artificial and spontaneous mutants of Strain FO-2546 which belong to Coelomycetes and have FO-2546 substance producing ability are usable in the present invention. Further, the mutated strain obtained by cell technology method such as cell fusion and gene manipulation technology which produce FO-2546 substance are included within the scope of the FO-2546 substance producing microorganism of the present invention.
In the present invention, a microorganism FO-2546 substance producing strain belonging to Coelomycetes is cultured in a nutrient medium. A culturing method generally used for fungi
can be used on the present invention. A nutrient medium containing assimilable carbon source, assimilable nitrogen source and, if required together with inorganic salt can be used. Examples of assimilable carbon sources are glucose, sucrose, molasses, dextrin or cellulose etc. and combinations thereof.
Examples of assimilable nitrogen sources are organic nitrogen source such as peptone, meat extract, yeast extract, dry yeast, soybean powder, corn steep liquor, cotton seed cake, casein, soybean protein hydrolysate, amino acid or urea, and inorganic nitrogen source such as ammonium salt, and combinations thereof. If required, inorganic salt such as sodium salt, potassium salt, calcium salt, magnesium salt and phosphate salt, and heavy metallic salt can be added to the medium. Further, trace nutrient element, growth stimulant or precusor can be added in the medium, if desired.
Cultivation should be performed under aerobic condition, e. g., by shaking or agitation. Submerged aeration agitation cultivation ispreferable for industrial production. Cultivation can be preferably performed at neutral pH. Cultivation temperature is usually at 20- 37βC. preferably at 24 - 30°C. FO-2546 substance is accumulated in the cultured mass by culturing for 2 - 3 days, and when maximum production of the FO-2546 substance is obtained, cultivation can be terminated.
These culturing condition such as medium composition, pH of the medium, culturing temperature, agitation time and aeration rate can be varied depending upon the microorganism strain used
and the atmospheric conditions for obtaining the preferable result. Anti-foaming agent such as silicon oil, vegetable oil or surface active agent can also be used if necessary. The FO- 2546 substance is accumulated in mycelia and in the cultured broth. The FO-2546 substance can preferably be isolated from supernatant and mycelia separated by centrifugation of cultured broth.
The FO-2546 substance can be extracted from supernatant with water-immiscible organic solvent such as ethyl acetate, butyl acetate or benzene, and crude FO-2546 substance can be obtained by concentrating the extract in vacuo. The crude substance is subjected to a known purification process for 1ipophilie substance, for example, by column chromatography using a carrier such as silica gel or alumina. Each component of the FO-2546 substance such as F0-2546A, -B, -C, -D, -I, -J and -K substance can be isolated.
The FO-2546 substance can also be isolated from mycelial cells by extracting the mycelia with water- iscible organic solvent such as aqueous acetone or aqueous methanol. The extract is concentrated under reduced pressure and the concentrate is subjected to further extraction with water-immiscible organic solvent such as ethyl acetate, butyl acetate or benzene. The extract which is optionally combined with the extract obtained from cultured filtrate hereinabove explained, can be purified, or it can be purified and separated by the same procedure as of the above described to obtained each component of the FO-2546 substance.
Physico-chemical properties:
Physico-chemical properies of F0-2546A substance, F0-2546B substance, F0-2546C substance. F0-2546D substance, F0-2546I substance, F0-2546J substance and F0-2546K substance are illustrated in Table 2 and Table 3.
Some properties of the F0-2546A substance, F0-2546B substance, F0-2546C substance, F0-2546D substance, F0-2546I substance, F0-2546J substance and F0-2546K substance are shown as follows.
Solubility in solvent : soluble in methanol, ethanol, aceto- nitrile, ethyl acetate and benzene; insoluble in water Nature of substance : neutral Appearance : yellowish powder Table 2
(To be continued Table 2)
Table 3
(To be continued Table 3)
Chemical structure F0-2546A substance :
HJ C CH,
F0-2546B substance
H3 C CH3
F0-2546D substance
H3 C CH3
F0-2546I substance
H3 C CH3
F0-2546J substance
H3 C CH3
F0-2546K substance
Biological properties: (1) Inhibitory action on rat acyl CoA cholesterol acyltransferase:
An inhibitory action of the FO-2546 substance for acyl CoA cholesterol acyltransferase is tested according to the method of Kyoda et al. (J. Antibiotics. 44: 136, 1991). A crude enzyme prepared from rat liver microsome fraction is added in the 100 mM phosphate buffer solution (pH 7.4) 200 #M containing 300 li M bovine serum albumin, 30 /zM Cl-1 C 3 oleoyl-CoA (0.02 zCi) and 30 #M cholesterol (solubilized with 1/30 w/w Triton WR-1339) and incubated at 37°C for 30 minutes. Total lipid is extracted
with a mixture of chloroform-methanol (2 : 1), then each lipid is separated by thin layer chromatography (Kiesel gel GF264, developer : petroleum ether-diethyl ether-acetic acid = 90 : 10: 1).
Radioactivity incorporated in the cholesterol ester fraction is analysed by RI scanner (Ambis Corp.) to assay an activity of acyl CoA cholesterol acyltransferase. The 50 % inhibition concentration on the enzyme is shown in Table 4. Table 4
Effect of the invention:
As illustrated hereinabove, FO-2546 substance and the individual components thereof, of the present invention show
» strong inhibitory action on acyl CoA cholesterol acyltransferase. Accordingly the substances are useful in the treatment of high serum cholesterol in mammals, including humans.
As used herein, treatment is meant to include both the prevention and alleviation of high serum cholesterol. The
substances may be administered to a subject in need of treatment by a variety of conventional routes of administration, including oral and parenteral. In general, these substances will be administered orally or parenterally at dosages between about 0.1 and about 10 mg/kg body weight of the subject to be treated per day, preferably from about 0.5 to 5 mg/kg. However, some variation in dosage will necessarily occur depending on the condition of the subject being treated and the activity of the compound being employed. The person responsible for administration will, in any event, determine the appropriate dose for the individual subject.
A substance of the invention may be administered alone or in combination with pharmaceutically acceptable carriers, in either single or multiple doses. Suitable pharmaceutically carriers include inert solid diluents or fillers, sterile aqueous solution and various organic solvents. The resulting pharmaceutical compositions are then readily administered in a variety of dosage forms such as tablets, powders, lozenges, syrups, injectable solutions and the like. These pharmaceutical compositions can, if desired, contain additional ingredients such as flavorings, binders, excipients and the like. Thus, for purposes of oral administration, tablets containing various excipients such as sodium citrate, calcium carbonate and calcium phosphate may be employed along with various disintegrants such as starch, alginic acid and certain complex silicates, together with binidng agents such as polyvinylpyrrolidone, sucrose, gelatin and acacia. Additionally, lubricating agents such as
magnesiu stearate, sodium lauryl sulfate and talc are often useful for tabletting purposes. Solid compositions of a similar type may also be employed as fillers in sort and hard filled gelatin capsules. Preferred materials for this include lactose or milk sugar and high molecular weight polyethylene glycols. When aqueous suspensions or elixirs are desired for oral administration, the essential active ingredient therein may be combined with various sweetening or flavoring agents coloring matter or dyes and. if desired, emulsifying or suspending agents, together with diluents such as water, ethanol, propylene glycol, glycerin and combinations thereof.
For parenteral administration, solutions of substance of the invention in seasa e or peanut oil, aqueous propylene glycol, or in sterile aqueous solution may be employed. Such aqueous solutions should be suitably buffered if necessary and the liquid diluent first rendered isotonic with sufficient saline or glucose. Such solutions are especially suitable for intravenous, intramuscular, subcutaneous and intraperitioneal administration.
Brief explanation of the drawings;
Figure 1 UV spectrum of F0-2546A substance, Figure 2 IR spectrum of F0-2546A substance, Figure 3 'H-NMR spectrum of F0-2546A substance, Figure 4 13C-NMR spectrum of F0-2546A substance, Figure 5 UV spectrum of F0-2546B substance, Figure 6 IR spectrum of F0-2546B substance, Figure 7 'H-NMR spectrum of F0-2546B substance. Figure 8 13C-NMR spectrum of F0-2546B substance.
Figure 9 : UV spectrum of F0-2546C substance, Figure 10 : IR spectrum of F0-2546C substance, Figure 11 : 'H-NMR spectrum of 2546C substance. Figure 12 : 13C-NMR spectrum of F0-2546C substance, Figure 13 : UV spectrum of F0-2546D substance. Figure 14 : IR spectrum of F0-2546D substance, Figure 15 : 'H-NMR spectrum of 2546D substance, Figure 16 : 13C-NMR spectrum of F0-2546D substance. Figure 17 : UV spectrum of F0-2546I substance, Figure 18 : IR spectrum of F0-2546I substance, Figure 19 : 'H-NMR spectrum of F0-2546I substance, Figure 20 : 13C-NMR spectrum of F0-2546I substance. Figure 21 : UV spectrum of F0-2546J substance, Figure 22 : IR spectrum of F0-2546J substance, Figure 23 : 'H-NMR spectrum of 2546J substance, Figure 24 : 13C-NMR spectrum of F0-2546J substance Figure 25 : UV spectrum of F0-2546K substance, Figure 26 : IR spectrum of F0-2546K substance, Figure 27 : 'H-NMR spectrum of 2546K substance and Figure 28 : 13C-NMR spectrum of F0-2546K substance. Following example illustrates the present invention but is not construed as limiting. Example 1.
A medium 100 ml (pH adjudted to 6.0) comprising glucose 21 yeast extract 0.21 MgS04 • 7H20 0.051 polypeptone 0.51 potasium dihydrogen phosphate 0.1% and agar 0.1 % in 500 ml Erlenmeyer flask with cotton seal was steam sterilized. One loopful
microorganism strain F0-2546 FERM BP-4406 grown on a nutrient agar medium was inoculated aseptically into the medium and shake cultured at 27°C for 48 hours to prepare seed culture liquid.
A medium comprising soluble starch 3. 0 %, glycerol 1.0 1 soybean powder 2.01 dry yeast 0.31 potasium chloride 0.31 calcium carbonate 0.21 MgS04 • 7H20 0.05 % and potasium dihydrogen phosphate 0.05 % (pH adjusted to 6.5) in 30 lit. jar- fer enter was sterilized and cooled. The seed culture 200 ml was transferred aseptically thereto and the inoculated medium was cultured aerobically at agitation 250 rp , aeration 10 lit./min. at 27°C for 72 hours. Cultured liquid 20 lit. including mycelia was extracted with ethyl acetate 18 lit. The extract was concentrated under reduced pressure to obtain crude substance 19g. The crude substance was charged on a column of silica gel (400g, Merk, Art 9385) and eluted with a mixture of chloroform- methanol (99 : 1). Active fractions containing FO-2546 substance comprising each 100 ml fraction were collected and concentrated in vacuo to obtain the crude substance 1.0 g.
The crude substance was purified separately divided with 10 times by high performance liquid chromatography (Trirota V, Nihon Bunko Co.. column YMC-Pack A-343, ODS resin. Yama ura Kagaku Kenkyusho) with linear gradient elution solvent sysytem of 75 - 90 % aqueous acetonitrile for 20 min., detection UV 225 nm, flow rate 8 ml/min. The F0-2546A substance 13 mg. F0-2546B substance 4.6 mg. F0-2546C substance 13 mg, F0-2546D substance 335 mg, F0-2546I substance 41 mg, F0-2546J substance 134 mg and F0-2546K substance 11 mg were obtained.
Claims
(1) A FO-2546 substance of the formula
wherein Ri is hydrogen or hydroxyl, and W is no bonding or oxygen, X is, together with Y, a group
O in which R2 is -CH=C (CH3)2 or -CH-C (CH3 ) _ and R3 is hydrogen or hydroxyl.
(2) A process for production of FO-2546 substance which comprises culturing FO-2546 substance producing microorganism belonging to
Coelomycetes in a nurtrient medium, accumulating the FO-2546 substance in the cultured mass, and isolating the FO-2546 substance therefrom.
(3) A process according to claim 2, wherein the FO-2546 substance producing microorganism is belonging to Coelomycetes is Strain FO-2546 FERM BP-4406.
(4) A microorganism strain belonging to Coelomycetes which has ability to produce FO-2546 substance.
(5) A microorganism strain according to claim 4, which is Strain FO-2546 FERM BP-4406.
(6) A F0-2546A substance of the formula
(7) A F0-2546B subs tance of the f ormu la
H3 C CH3
(8) A F0-2546D substance of the formula
H3 C CH3
(9) A F0-2546I subs tance of the formu la
H3 C CH3
(10) A F0-2546J substance of the formula
H3 C CH3
(11) A F0-2546K substance of the formula
H3 C CH3 (12) A pharmaceutical composition for the treatment of a disease attributed by accumulation of cholesterol in a mammalian subject which comprises a therapeutically effective amount of a compound of the formula
wherein Ri is hydrogen or hydroxyl, and W is no bonding or oxygen. X is, together with Y, a group
Ha C CHa
0
V in which R2 is -CH= C (CH3)2 or -C'H-t (CH3 )2 and R3 is hydrogen or hydroxyl, and a pharmaceutical acceptable carrier. (13) A method for the treatment of a disease attributed by accumulation of cholesterol in a mammalian subject, which comprises administering to said subject a therapeutically effective amount of a compound of the formula,
wherein Ki is hydrogen or hydroxyl, and W is no bonding or oxygen, X is, together with Yι a group
in which R2 is -CH = C (CH3)2
(CH3 ) _ and R3 is hydrogen or hydroxyl.
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP284760/92 | 1992-10-22 | ||
JP28476092 | 1992-10-22 | ||
PCT/JP1993/001518 WO1994009004A1 (en) | 1992-10-22 | 1993-10-21 | Fo-2546 substance, its production and its use as inhibitor of acyl-coa cholesterol acyltransferase |
Publications (1)
Publication Number | Publication Date |
---|---|
EP0665845A1 true EP0665845A1 (en) | 1995-08-09 |
Family
ID=17682657
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
EP93923038A Withdrawn EP0665845A1 (en) | 1992-10-22 | 1993-10-21 | Fo-2546 substance, its production and its use as inhibitor of acyl-coa cholesterol acyltransferase |
Country Status (2)
Country | Link |
---|---|
EP (1) | EP0665845A1 (en) |
WO (1) | WO1994009004A1 (en) |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5541208A (en) * | 1994-01-24 | 1996-07-30 | Merck & Co., Inc. | Indole diterpene alkaloid compounds |
-
1993
- 1993-10-21 WO PCT/JP1993/001518 patent/WO1994009004A1/en not_active Application Discontinuation
- 1993-10-21 EP EP93923038A patent/EP0665845A1/en not_active Withdrawn
Non-Patent Citations (1)
Title |
---|
See references of WO9409004A1 * |
Also Published As
Publication number | Publication date |
---|---|
WO1994009004A1 (en) | 1994-04-28 |
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