CS271347B2 - Method of 4-androstene-3,17-dione and 1,4-androstadiene-3,17-diene production - Google Patents
Method of 4-androstene-3,17-dione and 1,4-androstadiene-3,17-diene production Download PDFInfo
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- CS271347B2 CS271347B2 CS869282A CS928286A CS271347B2 CS 271347 B2 CS271347 B2 CS 271347B2 CS 869282 A CS869282 A CS 869282A CS 928286 A CS928286 A CS 928286A CS 271347 B2 CS271347 B2 CS 271347B2
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- dione
- androstadiene
- nrrl
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- mycobacterium
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- 238000000034 method Methods 0.000 title claims abstract description 13
- AEMFNILZOJDQLW-QAGGRKNESA-N androst-4-ene-3,17-dione Chemical compound O=C1CC[C@]2(C)[C@H]3CC[C@](C)(C(CC4)=O)[C@@H]4[C@@H]3CCC2=C1 AEMFNILZOJDQLW-QAGGRKNESA-N 0.000 title claims abstract description 11
- 238000004519 manufacturing process Methods 0.000 title claims abstract description 4
- 229960005471 androstenedione Drugs 0.000 title claims description 9
- AEMFNILZOJDQLW-UHFFFAOYSA-N androstenedione Natural products O=C1CCC2(C)C3CCC(C)(C(CC4)=O)C4C3CCC2=C1 AEMFNILZOJDQLW-UHFFFAOYSA-N 0.000 title claims description 9
- 244000005700 microbiome Species 0.000 claims abstract description 15
- LUJVUUWNAPIQQI-UHFFFAOYSA-N (+)-androsta-1,4-diene-3,17-dione Natural products O=C1C=CC2(C)C3CCC(C)(C(CC4)=O)C4C3CCC2=C1 LUJVUUWNAPIQQI-UHFFFAOYSA-N 0.000 claims description 9
- LUJVUUWNAPIQQI-QAGGRKNESA-N androsta-1,4-diene-3,17-dione Chemical compound O=C1C=C[C@]2(C)[C@H]3CC[C@](C)(C(CC4)=O)[C@@H]4[C@@H]3CCC2=C1 LUJVUUWNAPIQQI-QAGGRKNESA-N 0.000 claims description 9
- 125000002328 sterol group Chemical group 0.000 claims description 7
- 241000186359 Mycobacterium Species 0.000 claims description 5
- 238000002360 preparation method Methods 0.000 claims description 5
- 230000000593 degrading effect Effects 0.000 claims description 4
- -1 lnnn-5 Chemical compound 0.000 claims description 2
- OSVMTWJCGUFAOD-KZQROQTASA-N formestane Chemical compound O=C1CC[C@]2(C)[C@H]3CC[C@](C)(C(CC4)=O)[C@@H]4[C@@H]3CCC2=C1O OSVMTWJCGUFAOD-KZQROQTASA-N 0.000 claims 3
- DCIFNKCWKJGATM-CDJYOOERSA-N (9r,10s,13s)-10,13-dimethyl-2,4,5,6,7,9,11,12-octahydro-1h-cyclopenta[a]phenanthrene-3,17-dione Chemical compound C1C(=O)CC[C@]2(C)[C@H]3CC[C@](C)(C(C=C4)=O)C4=C3CCC21 DCIFNKCWKJGATM-CDJYOOERSA-N 0.000 abstract 1
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 description 9
- 239000000758 substrate Substances 0.000 description 8
- 229950005143 sitosterol Drugs 0.000 description 7
- 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
- 241000187488 Mycobacterium sp. Species 0.000 description 6
- 230000015556 catabolic process Effects 0.000 description 6
- 238000006731 degradation reaction Methods 0.000 description 6
- 239000003995 emulsifying agent Substances 0.000 description 6
- 238000000855 fermentation Methods 0.000 description 6
- 230000004151 fermentation Effects 0.000 description 6
- LPZCCMIISIBREI-MTFRKTCUSA-N Citrostadienol Natural products CC=C(CC[C@@H](C)[C@H]1CC[C@H]2C3=CC[C@H]4[C@H](C)[C@@H](O)CC[C@]4(C)[C@H]3CC[C@]12C)C(C)C LPZCCMIISIBREI-MTFRKTCUSA-N 0.000 description 5
- 241000187481 Mycobacterium phlei Species 0.000 description 5
- 229940055036 mycobacterium phlei Drugs 0.000 description 5
- 150000003431 steroids Chemical class 0.000 description 5
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 4
- LGJMUZUPVCAVPU-UHFFFAOYSA-N beta-Sitostanol Natural products C1CC2CC(O)CCC2(C)C2C1C1CCC(C(C)CCC(CC)C(C)C)C1(C)CC2 LGJMUZUPVCAVPU-UHFFFAOYSA-N 0.000 description 4
- NJKOMDUNNDKEAI-UHFFFAOYSA-N beta-sitosterol Natural products CCC(CCC(C)C1CCC2(C)C3CC=C4CC(O)CCC4C3CCC12C)C(C)C NJKOMDUNNDKEAI-UHFFFAOYSA-N 0.000 description 4
- KZJWDPNRJALLNS-VJSFXXLFSA-N sitosterol 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)CC[C@@H](CC)C(C)C)[C@@]1(C)CC2 KZJWDPNRJALLNS-VJSFXXLFSA-N 0.000 description 4
- KZJWDPNRJALLNS-VPUBHVLGSA-N (-)-beta-Sitosterol Natural products O[C@@H]1CC=2[C@@](C)([C@@H]3[C@H]([C@H]4[C@@](C)([C@H]([C@H](CC[C@@H](C(C)C)CC)C)CC4)CC3)CC=2)CC1 KZJWDPNRJALLNS-VPUBHVLGSA-N 0.000 description 3
- CSVWWLUMXNHWSU-UHFFFAOYSA-N (22E)-(24xi)-24-ethyl-5alpha-cholest-22-en-3beta-ol Natural products C1CC2CC(O)CCC2(C)C2C1C1CCC(C(C)C=CC(CC)C(C)C)C1(C)CC2 CSVWWLUMXNHWSU-UHFFFAOYSA-N 0.000 description 3
- KLEXDBGYSOIREE-UHFFFAOYSA-N 24xi-n-propylcholesterol Natural products C1C=C2CC(O)CCC2(C)C2C1C1CCC(C(C)CCC(CCC)C(C)C)C1(C)CC2 KLEXDBGYSOIREE-UHFFFAOYSA-N 0.000 description 3
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 3
- 241000193830 Bacillus <bacterium> Species 0.000 description 3
- ARVGMISWLZPBCH-UHFFFAOYSA-N Dehydro-beta-sitosterol Natural products C1C(O)CCC2(C)C(CCC3(C(C(C)CCC(CC)C(C)C)CCC33)C)C3=CC=C21 ARVGMISWLZPBCH-UHFFFAOYSA-N 0.000 description 3
- 241000186365 Mycobacterium fortuitum Species 0.000 description 3
- 241000187480 Mycobacterium smegmatis Species 0.000 description 3
- 241000586779 Protaminobacter Species 0.000 description 3
- 241000187747 Streptomyces Species 0.000 description 3
- MJVXAPPOFPTTCA-UHFFFAOYSA-N beta-Sistosterol Natural products CCC(CCC(C)C1CCC2C3CC=C4C(C)C(O)CCC4(C)C3CCC12C)C(C)C MJVXAPPOFPTTCA-UHFFFAOYSA-N 0.000 description 3
- 230000015572 biosynthetic process Effects 0.000 description 3
- 150000001875 compounds Chemical class 0.000 description 3
- 239000000543 intermediate Substances 0.000 description 3
- 229920000136 polysorbate Polymers 0.000 description 3
- 235000015500 sitosterol Nutrition 0.000 description 3
- NLQLSVXGSXCXFE-UHFFFAOYSA-N sitosterol Natural products CC=C(/CCC(C)C1CC2C3=CCC4C(C)C(O)CCC4(C)C3CCC2(C)C1)C(C)C NLQLSVXGSXCXFE-UHFFFAOYSA-N 0.000 description 3
- 239000002904 solvent Substances 0.000 description 3
- 238000003786 synthesis reaction Methods 0.000 description 3
- LPZCCMIISIBREI-JXMPMKKESA-N (Z)-24-ethylidenelophenol Chemical compound C[C@@H]1[C@@H](O)CC[C@]2(C)[C@@H](CC[C@@]3([C@@H]([C@H](C)CC/C(=C/C)C(C)C)CC[C@H]33)C)C3=CC[C@H]21 LPZCCMIISIBREI-JXMPMKKESA-N 0.000 description 2
- XNWFRZJHXBZDAG-UHFFFAOYSA-N 2-METHOXYETHANOL Chemical compound COCCO XNWFRZJHXBZDAG-UHFFFAOYSA-N 0.000 description 2
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 2
- 241001467578 Microbacterium Species 0.000 description 2
- 229930182558 Sterol Natural products 0.000 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 description 2
- 239000003112 inhibitor Substances 0.000 description 2
- 150000003432 sterols Chemical class 0.000 description 2
- 235000003702 sterols Nutrition 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- WSLDOOZREJYCGB-UHFFFAOYSA-N 1,2-Dichloroethane Chemical compound ClCCCl WSLDOOZREJYCGB-UHFFFAOYSA-N 0.000 description 1
- VBICKXHEKHSIBG-UHFFFAOYSA-N 1-monostearoylglycerol Chemical compound CCCCCCCCCCCCCCCCCC(=O)OCC(O)CO VBICKXHEKHSIBG-UHFFFAOYSA-N 0.000 description 1
- 241000186063 Arthrobacter Species 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 241000186146 Brevibacterium Species 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- ZAFNJMIOTHYJRJ-UHFFFAOYSA-N Diisopropyl ether Chemical compound CC(C)OC(C)C ZAFNJMIOTHYJRJ-UHFFFAOYSA-N 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- IAYPIBMASNFSPL-UHFFFAOYSA-N Ethylene oxide Chemical class C1CO1 IAYPIBMASNFSPL-UHFFFAOYSA-N 0.000 description 1
- 241000186367 Mycobacterium avium Species 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-L Phosphate ion(2-) Chemical compound OP([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-L 0.000 description 1
- 235000019764 Soybean Meal Nutrition 0.000 description 1
- 240000008042 Zea mays Species 0.000 description 1
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 1
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 1
- 238000005273 aeration Methods 0.000 description 1
- 229940076810 beta sitosterol Drugs 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 229940041514 candida albicans extract Drugs 0.000 description 1
- 239000001569 carbon dioxide Substances 0.000 description 1
- 229910002092 carbon dioxide Inorganic materials 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 235000012000 cholesterol Nutrition 0.000 description 1
- 235000005822 corn Nutrition 0.000 description 1
- 235000014113 dietary fatty acids Nutrition 0.000 description 1
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 description 1
- 238000004945 emulsification Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 239000000194 fatty acid Substances 0.000 description 1
- 229930195729 fatty acid Natural products 0.000 description 1
- 238000005805 hydroxylation reaction Methods 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- VUZPPFZMUPKLLV-UHFFFAOYSA-N methane;hydrate Chemical compound C.O VUZPPFZMUPKLLV-UHFFFAOYSA-N 0.000 description 1
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 1
- 230000002906 microbiologic effect Effects 0.000 description 1
- 238000003798 microbiological reaction Methods 0.000 description 1
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 1
- 235000019796 monopotassium phosphate Nutrition 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 description 1
- 229920000151 polyglycol Polymers 0.000 description 1
- 239000010695 polyglycol Substances 0.000 description 1
- 238000001953 recrystallisation Methods 0.000 description 1
- 238000010898 silica gel chromatography Methods 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 239000004455 soybean meal Substances 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- 238000000844 transformation Methods 0.000 description 1
- 239000000080 wetting agent Substances 0.000 description 1
- 238000010626 work up procedure Methods 0.000 description 1
- 239000012138 yeast extract Substances 0.000 description 1
Classifications
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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
- C12P33/00—Preparation of steroids
- C12P33/005—Degradation of the lateral chains at position 17
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- Organic Chemistry (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Life Sciences & Earth Sciences (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Health & Medical Sciences (AREA)
- General Engineering & Computer Science (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Biotechnology (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Microbiology (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Steroid Compounds (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
Description
Způsob výroby 4-androsten-3,17-dionu a 1,4-androstadien-3,17-dionuProcess for preparing 4-androstene-3,17-dione and 1,4-androstadiene-3,17-dione
Způsob výroby 4-androsten-3,17-dionu a 1,4-androstadien-3,17-dionu obecného vzorce I, ve kterém ..... znamená jednoduchou vazbu nebo dvojnou vazbu, vyznačující se tím, že se -sitosterol vzorce II fermentuje kulturou kmene mikroorganismů schopných odbourávat postranní řetězce sterolů. Vyráběné sloučeniny se používají jako meziprodukty к syntéze farmakologicky účinných steroidů.A process for the preparation of 4-androstene-3,17-dione and 1,4-androstadiene-3,17-dione of the general formula I, in which it is a single bond or a double bond, characterized in that the isosterol is of the formula II fermentes with a culture of a strain of microorganisms capable of degrading the sterol side chains. The compounds produced are used as intermediates for the synthesis of pharmacologically active steroids.
Vynález se týká způsobu výroby 4-androsten-3,17-dionu a 1,4-androstadien-3,17-dionu. Tyto sloučeniny se používají к syntéze farmakologicky účinných steroidů.The invention relates to a process for the preparation of 4-androstene-3,17-dione and 1,4-androstadiene-3,17-dione. These compounds are used for the synthesis of pharmacologically active steroids.
Předmětem předloženého vynálezu je způsob výroby 4-androsten-3,17-dionu a 1,4-androstaďien-3,17-dionu obecného vzorce IThe present invention provides a process for the preparation of 4-androstene-3,17-dione and 1,4-androstadiene-3,17-dione of formula I
ve kterém , , , , , znamená jednoduchou nebo dvojnou vazbu, který spočívá v tom, vzorce II že se -sitosterolwherein,,,,, means a single or double bond consisting in that Formula II is -sitosterol
fermentuje kulturou kmene mikroorganismů, který je schopen odbourávat postranní řetězce sterilů.fermented by culture of a strain of microorganisms capable of breaking down the sterile side chains.
Výhodně se postup podle vynálezu provádí tak, že se oč-sitosterol vzorce II fermen- <Preferably, the process according to the invention is carried out by the use of acetosterosterol of formula (II)
tuje kmenem mikroorganismu, které jsou schopny odbourávat postranní řetězce sterolů rodu Mycobacterium nebo Norcardia.It is a strain of microorganisms capable of degrading the sterol side chains of Mycobacterium or Norcardia.
Výhodné provedení postupu podle vynálezu spočívá dále v tom, že se -si tosterol vzorce II fermentuje pomocí mikroorganismu Mycobacterium spec. NRRL Β-3ΘΟ5, Mycobacterium spec. NRRL B—3683, Mycobacterium phleí NRRL B-8154 nebo Mycobacterium fortuitum NRRL N-8153.A preferred embodiment of the process according to the invention is further characterized in that the tosterol of the formula II is fermented with the microorganism Mycobacterium spec. NRRL Β -3-5, Mycobacterium spec. NRRL B-3683, Mycobacterium phlei NRRL B-8154 or Mycobacterium fortuitum NRRL N-8153.
Je známo, že četné mikroorganismy (například mikroorganismy rodů Arthrobacter, Brevibacterium, Microbacterium, Protaminobacter, Bacillus, Norcardia, Streptomyces a zejména Mycobacterium) mají přirozenou schopnost odbourávat v přírodě se vyskytujícíNumerous microorganisms (e.g., microorganisms of the genera Arthrobacter, Brevibacterium, Microbacterium, Protaminobacter, Bacillus, Norcardia, Streptomyces, and especially Mycobacterium) are known to have a natural ability to degrade naturally occurring
3/З-hydroxy-A5-steroly (jako cholesterol nebo sitosterol) na oxid uhličitý a vodu, přičemž se při tomto odbourávání intermediárně tvoří 4-androsten-3,17-dion a 1,4-androstadien-3,17-dion.3/2-hydroxy- 5 5 -sterols (such as cholesterol or sitosterol) to carbon dioxide and water, the intermediate being 4-androstene-3,17-dione and 1,4-androstadiene-3,17-dione .
Oále je známo, že pomocí přísad inhibitorů nebo mutantů mikroorganismů je možno řídit odbourávání sterolu tak, že se zamezí odbourávání vzniklého 4-androsten-3,17-dionu nebo 1,4-androstadien-3,17-dionu (srov. německé zveřejněné spisy 15 43 269 a 15 93 327, jakož i americký patentový spis 3 684 657).It is further known that the addition of inhibitors or mutants of microorganisms can control the degradation of sterol by preventing the degradation of the resulting 4-androstene-3,17-dione or 1,4-androstadiene-3,17-dione (cf. German published publications). 15 43 269 and 15 93 327 as well as U.S. Pat. No. 3,684,657).
Pro odborníka je tudíž překvapující, že se za známých podmínek odbourává také postraní řetězec -sitosterolu, vzhledem к tomu, že je známo, že odbourávání postranních řetězců sterolů způsobuje velmi komplexní fermentační systém a nebylo tudíž možno očekávat, že všechny enzymy spolupůsobící při odbourávání postranních řetězců přírodních steroidů mají schopnost způsobit také odbourání postranních řetězců této sloučeniny. Kromě toho nebylo možno předpokládat, že se při tomto odbourávání hydrogenuje Δ^-dvojná vazba cL-sitosterolu a že se odštěpuje methylová skupina v poloze 4 Z*.It is therefore surprising for the skilled person that under the known conditions the side chain of -sitosterol also degrades, since it is known that the degradation of the sterol side chains causes a very complex fermentation system and it was therefore not expected that all enzymes involved in the side chain degradation Natural steroids also have the ability to cause side chain degradation of this compound. In addition, it could not be assumed that the β-double bond of α-sitosterol was hydrogenated in this degradation and that the methyl group at the 4-position of the Z @ 2 was cleaved.
Nehledě na použití jiných výchozích látek se postup podle vynálezu provádí za stejných fermentačních podmínek, které se používají také při známých mikrobiologických reak* cích sterolů к odbourání postranních řetězců.Apart from the use of other starting materials, the process according to the invention is carried out under the same fermentation conditions which are also used in the known microbiological reaction of sterols to degrade side chains.
Podle tohoto vynálezu se fermentace provádí za použití kultury mikroorganismu, která se obvykle používá к odbourávání postranních řetězců sterolů. Vhodnými kulturami jsou například kultury bakterií rodů Arhtrobacter, Bervibacterium, Microbacterium, Protaminobacter, Streptomyces nebo zejména rodů Mycobacterium, schopné odbourávání postranních řetězců sterolů. Jako vhodné mikroorganismy lze uvést například: Mycrobacterium lactum IAM-1640, Protaminobacter alboflavus IAM-1040, Bacillus roseus IAM-1257, Bacillus spharicus ATTC-7055, Norcardia gardneri IAM-105, Norcardia minima IAM-374, Norcardia cirallina IFO-3338, Streptomyces rubescens IAM-74 nebo zejména mikroorganismy Mycobacterium avium IF0-3082, Mycobacterium phlei IFO-3158, Mycobacterium phlei (Institut fiir Gesundheitswesen, Budapest Č. 29), Mycobacterium phlei ATCC-354, Mycobacterium smegmatis IFO-3084, Mycobacterium smegmatis ATTC-20, Mycobacterium smegmatis (Institut fur Gesundheitswesen, Budapest č. 27), Mycobacterium smegmatis ATCC-19979 a Mycobacterium fortuitum CBS-49566.According to the present invention, the fermentation is carried out using a culture of a microorganism which is usually used to break down the sterol side chains. Suitable cultures are, for example, cultures of bacteria of the genera Arhtrobacter, Bervibacterium, Microbacterium, Protaminobacter, Streptomyces or in particular of the genera Mycobacterium, capable of degrading the sterol side chains. Suitable microorganisms include, for example: Mycrobacterium lactum IAM-1640, Protaminobacter alboflavus IAM-1040, Bacillus roseus IAM-1257, Bacillus spharicus ATTC-7055, Norcardia gardneri IAM-105, Norcardia minima IAM-374, Norcardia cirallina IFO-3338, Streptomyces rubescens IAM-74 or especially microorganisms Mycobacterium avium IF0-3082, Mycobacterium phlei IFO-3158, Mycobacterium phlei (Institut für Gesundheitswesen, Budapest No. 29), Mycobacterium phlei ATCC-354, Mycobacterium smegmatis IFO-3084, 20 Mycobacterium smegmatis (Institut fur Gesundheitswesen, Budapest No. 27), Mycobacterium smegmatis ATCC-19979 and Mycobacterium fortuitum CBS-49566.
Zvláště vhodné jsou mikroorganismy Mycobacterium spec. NRRL B-3805, Mycobacterium spec. NRRL-3683, Mycobacterium phlei NRRL B-8154 a Mycobacterium fortuitum NRRL Β-Θ153, pomocí kterých je možno provádět fermantaci -sitosterolu bez použití přídavných inhibitorů inhibujících 9«C-hydroxylaci.Particularly suitable are Mycobacterium spec. NRRL B-3805, Mycobacterium spec. NRRL-3683, Mycobacterium phlei NRRL B-8154 and Mycobacterium fortuitum NRRL Β-Θ153, which can be used to fermantize sitosterol without the use of additional inhibitors of 9? -Hydroxylation.
Kultivace se provádí za podmínek obvykle používaných pro kultivaci těchto mikroorganismů ve vhodných živných půdách za provzdušňování. Poté se ke kulturám přidá substrát (rozpuštěný nebo výhodně emulgovaný ve vhodném rozpouštědle) a provádí se fermentace až se dosáhne maximální přeměny substrátu.The cultivation is carried out under the conditions usually used to cultivate these microorganisms in suitable nutrient media under aeration. Subsequently, a substrate (dissolved or preferably emulsified in a suitable solvent) is added to the cultures and fermentation is performed until the maximum conversion of the substrate is achieved.
Vhodnými rozpouštědly pro substrát jsou například methanol, ethanol, glykolmonomethylether, dimethylformamid nebo dimethylsulfoxid. Emulgace substrátu lze dosáhnout například tím, že se tento substrát v mikronisované formě nebo (rozpuštěn) v rozpouštědle mísitelném ί s vodou (jako v methanolu, ethanolu, acetonu, glykolmonomethyletheru, dimethylformamidu nebo dimethylsulfoxidu) vstřikuje pomocí trysky za silné turbulence do vody (výhodně zbavené vápníku), která obsahuje obvyklé pomocné emulgátory. Vhodnými pomocnými emulgátory ’ jsou neionogenní emulgátory, jako například adukty ethylenoxidu nebo esterů mastných kyselin polyglykolů. Jako vhodné emulgátory lze uvést například na trhu obvyklá smáčedla Tegin Tween^R) a SpánSuitable solvents for the substrate are, for example, methanol, ethanol, glycol monomethyl ether, dimethylformamide or dimethylsulfoxide. The emulsification of the substrate can be achieved, for example, by injecting the substrate in micronized form or (dissolved) in a water-miscible solvent (such as methanol, ethanol, acetone, glycol monomethyl ether, dimethylformamide or dimethylsulfoxide) by injection into a water (preferably freed) calcium), which contains conventional auxiliary emulsifiers. Suitable auxiliary emulsifiers are nonionic emulsifiers, such as ethylene oxide adducts or polyglycol fatty acid esters. Suitable emulsifiers include, for example, the commercially available wetting agents Tegin Tween (R) and Span
Optimální koncentrace substrátu, doba přidávání substrátu a doba trvání fermentace závisí na struktuře použitého eubstnátu a na druhu použitého mikroorganismu. Tyto veličiny se musí, jak je to u mikrobiologických přeměn steroidů obecně zapotřebí, zjistit v jednotlivém případě předběžnými pokusy, které jsou odborníkovi běžné.The optimum substrate concentration, substrate addition time, and fermentation duration depend on the structure of the substrate used and the type of microorganism used. These quantities must, as is generally necessary for microbiological transformations of steroids, be determined on a case-by-case basis by preliminary experiments customary to one skilled in the art.
Deriváty 4-androsten-3,17-dionu obecného vzorce I, které lze vyrobit postupem podle vynálezu jsou jak známo cennými meziprodukty používanými к syntéze farmakologicky účinných steroidů.The 4-androstene-3,17-dione derivatives of the formula I which can be prepared by the process according to the invention are known as valuable intermediates used for the synthesis of pharmacologically active steroids.
Následující příklady slouží к bližšímu objasnění postupu podle vynálezu.The following examples serve to illustrate the process of the invention in more detail.
Příklad 1Example 1
Obsah Elenmeyerovy baňky o obsahu 2 litrů s 500 ml sterilní živné půdy obsahující % extraktu z kvasnicContent of a 2 liter Elenmeyer flask with 500 ml of sterile broth containing% yeast extract
0,45 % dinatriumhydrogenfosfátu0.45% disodium hydrogen phosphate
0,34 % kaliumdihydrogenfosfátu a0,34% of potassium dihydrogen phosphate and
0,20 % emulgátoru Tween^) 80 a upravené na pH 6,7 se naočkuje suspenzí suché kultury Mycobacterium spec. NRRL B-3805 a protřepává se po dobu 3 dnů při teplotě 30 °C a 190 otáčkách za minutu.0.20% of Tween® 80 emulsifier and adjusted to pH 6.7 were seeded with a dry culture suspension of Mycobacterium spec. NRRL B-3805 and shaken for 3 days at 30 ° C and 190 rpm.
Erlenmeyerových baněk (500 ml) vždy se 100 ml sterilní živné půdy obsahujícíErlenmeyer flasks (500 ml) with 100 ml sterile broth each
2,5 % kukuřičného výluhu2.5% corn steep liquor
0,3 % dinamoniumhydrogenfosfátu0.3% dinammonium hydrogen phosphate
0,25 % sojové moučky0.25% soya meal
0,25 % emulgátoru Tween ^) 80 a upravené na pH 7,0 se naočkuje vždy 5 ml kultury Mycobacterium spec, a obsah baněk se protřepává 24 hodin při teplotě 30 °C a 220 otáčkách za minutu. Potom se ke každé kultuře přidá roztok 100 mg aC -sitosterolu v 3,0 ml dimethylformamidu a fermantace se provádí dalších 96 hodin při teplotě 30 °C.0.25% of Tween® 80 emulsifier, adjusted to pH 7.0, are inoculated with 5 ml of a Mycobacterium spec culture, and the flasks are shaken for 24 hours at 30 ° C and 220 rpm. A solution of 100 mg of α-sitosterol in 3.0 ml of dimethylformamide was then added to each culture and the fermentation was continued for another 96 hours at 30 ° C.
Spojené kultury se extrahují ethylenchloridem, extrakt se zahustí ve vakuu, zbytek se čistí chromatografií na sloupci silikagelu a po překrystalování z diisopropyletheru se získá 160 mg 4-androsten-3,17-dionu.The combined cultures were extracted with ethylene chloride, the extract was concentrated in vacuo, and the residue was purified by silica gel column chromatography to give 160 mg of 4-androstene-3,17-dione after recrystallization from diisopropyl ether.
Kromě toho se získá zpět 385 mg nezreagovaného eč-sitosterolu.In addition, 385 mg of unreacted β-sitosterol is recovered.
Příklad 2Example 2
Za podmínek popsaných v příkladu 1, avšak zapoužití Mycobacterium spec. NRRL B-3683 se nechá zreagovat celkem 1 000 mg -sitosterolu. Po obvyklém zpracování se získá vedle 630 mg nezreagovaného «č -sitosterolu 140 mg 1,4-androstadien-3,17-dionu.Under the conditions described in Example 1 but using Mycobacterium spec. NRRL B-3683 was reacted with a total of 1000 mg of sitosterol. After usual work-up, in addition to 630 mg of unreacted N-sitosterol, 140 mg of 1,4-androstadiene-3,17-dione is obtained.
PŘEDMĚT VYNÁLEZUSUBJECT OF THE INVENTION
Claims (3)
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DE19853544662 DE3544662A1 (en) | 1985-12-13 | 1985-12-13 | METHOD FOR PRODUCING 4-ANDROSTEN-3,17-DION AND 1,4-ANDROSTADIEN-3,17-DION |
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| Publication Number | Publication Date |
|---|---|
| CS928286A2 CS928286A2 (en) | 1990-02-12 |
| CS271347B2 true CS271347B2 (en) | 1990-09-12 |
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CS869282A CS271347B2 (en) | 1985-12-13 | 1986-12-12 | Method of 4-androstene-3,17-dione and 1,4-androstadiene-3,17-diene production |
Country Status (17)
| Country | Link |
|---|---|
| EP (1) | EP0227588B1 (en) |
| JP (1) | JPH0720439B2 (en) |
| AT (1) | ATE69268T1 (en) |
| AU (1) | AU6772887A (en) |
| BG (1) | BG50613A3 (en) |
| CA (1) | CA1302923C (en) |
| CS (1) | CS271347B2 (en) |
| DD (1) | DD254025A5 (en) |
| DE (2) | DE3544662A1 (en) |
| DK (1) | DK419587D0 (en) |
| ES (1) | ES2038601T3 (en) |
| FI (1) | FI873489L (en) |
| GR (1) | GR3003640T3 (en) |
| HU (1) | HUT44082A (en) |
| NO (1) | NO873381D0 (en) |
| SU (1) | SU1679977A3 (en) |
| WO (1) | WO1987003620A1 (en) |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| US6071714A (en) | 1998-03-26 | 2000-06-06 | Forbes Medi-Tech, Inc. | Process for the microbial conversion of phytosterols to androstenedione and androstadienedione |
| KR100422161B1 (en) | 2001-05-11 | 2004-03-10 | (주)유진사이언스 | A method for preparation of Androst-4-ene-3,17-dione and Androsta-1,4-diene-3,17-dione |
Family Cites Families (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS4826549B1 (en) * | 1968-11-14 | 1973-08-11 | ||
| US3684657A (en) * | 1970-05-11 | 1972-08-15 | Searle & Co | Selective microbiological degradation of steroidal 17-alkyls |
| US3759791A (en) * | 1970-12-10 | 1973-09-18 | Searle & Co | Selective microbiological preparation of androst-4-ene-3,17-dione |
| JPS512670Y2 (en) * | 1971-06-28 | 1976-01-26 | ||
| US4293645A (en) * | 1976-03-01 | 1981-10-06 | The Upjohn Company | Process for preparing androsta-1,4-diene-3,17-dione and androst-4-ene-3,17-dione |
| DE2703645C3 (en) * | 1976-03-01 | 1981-11-12 | The Upjohn Co., 49001 Kalamazoo, Mich. | Process for the preparation of a mixture of androsta-1,4-diene-3,17-dione and androst-4-ene-3,17-dione |
| JPS52115548A (en) * | 1976-03-24 | 1977-09-28 | Matsushita Electric Ind Co Ltd | Heat pump type air conditioning system |
-
1985
- 1985-12-13 DE DE19853544662 patent/DE3544662A1/en not_active Withdrawn
-
1986
- 1986-12-09 HU HU86458A patent/HUT44082A/en unknown
- 1986-12-09 EP EP86730207A patent/EP0227588B1/en not_active Expired - Lifetime
- 1986-12-09 WO PCT/DE1986/000508 patent/WO1987003620A1/en active Application Filing
- 1986-12-09 AU AU67728/87A patent/AU6772887A/en not_active Abandoned
- 1986-12-09 DE DE86730207T patent/DE3682379D1/de not_active Expired - Fee Related
- 1986-12-09 AT AT86730207T patent/ATE69268T1/en not_active IP Right Cessation
- 1986-12-09 ES ES198686730207T patent/ES2038601T3/en not_active Expired - Lifetime
- 1986-12-09 FI FI873489A patent/FI873489L/en not_active IP Right Cessation
- 1986-12-09 JP JP62500101A patent/JPH0720439B2/en not_active Expired - Lifetime
- 1986-12-12 DD DD86297530A patent/DD254025A5/en not_active IP Right Cessation
- 1986-12-12 CA CA000525244A patent/CA1302923C/en not_active Expired - Lifetime
- 1986-12-12 CS CS869282A patent/CS271347B2/en not_active IP Right Cessation
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1987
- 1987-07-07 BG BG080473A patent/BG50613A3/en unknown
- 1987-08-10 SU SU4203113A patent/SU1679977A3/en active
- 1987-08-12 DK DK419587A patent/DK419587D0/en not_active Application Discontinuation
- 1987-08-12 NO NO873381A patent/NO873381D0/en unknown
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1992
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Also Published As
| Publication number | Publication date |
|---|---|
| DK419587A (en) | 1987-08-12 |
| GR3003640T3 (en) | 1993-03-16 |
| ATE69268T1 (en) | 1991-11-15 |
| CS928286A2 (en) | 1990-02-12 |
| FI873489A7 (en) | 1987-08-11 |
| FI873489A0 (en) | 1987-08-11 |
| DK419587D0 (en) | 1987-08-12 |
| ES2038601T3 (en) | 1993-08-01 |
| NO873381L (en) | 1987-08-12 |
| WO1987003620A1 (en) | 1987-06-18 |
| NO873381D0 (en) | 1987-08-12 |
| JPS63501922A (en) | 1988-08-04 |
| DD254025A5 (en) | 1988-02-10 |
| CA1302923C (en) | 1992-06-09 |
| HUT44082A (en) | 1988-01-28 |
| FI873489L (en) | 1987-08-11 |
| BG50613A3 (en) | 1992-09-15 |
| DE3682379D1 (en) | 1991-12-12 |
| DE3544662A1 (en) | 1987-06-19 |
| EP0227588A1 (en) | 1987-07-01 |
| SU1679977A3 (en) | 1991-09-23 |
| EP0227588B1 (en) | 1991-11-06 |
| JPH0720439B2 (en) | 1995-03-08 |
| AU6772887A (en) | 1987-06-30 |
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| IF00 | In force as of 2000-06-30 in czech republic | ||
| MK4A | Patent expired |
Effective date: 20011212 |