US3616226A - Process for producing unsaturated steroids - Google Patents

Process for producing unsaturated steroids Download PDF

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Publication number
US3616226A
US3616226A US746882A US3616226DA US3616226A US 3616226 A US3616226 A US 3616226A US 746882 A US746882 A US 746882A US 3616226D A US3616226D A US 3616226DA US 3616226 A US3616226 A US 3616226A
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ifo
secogona
compound
nucleus
carbon
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Expired - Lifetime
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US746882A
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English (en)
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Masao Isono
Takeshi Takahashi
Yoshio Yamasaki
Takuichi Miki
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Takeda Pharmaceutical Co Ltd
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Takeda Chemical Industries Ltd
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C49/00—Ketones; Ketenes; Dimeric ketenes; Ketonic chelates
    • C07C49/587—Unsaturated compounds containing a keto groups being part of a ring
    • C07C49/753—Unsaturated compounds containing a keto groups being part of a ring containing ether groups, groups, groups, or groups
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
    • Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10S435/00—Chemistry: molecular biology and microbiology
    • Y10S435/8215—Microorganisms
    • Y10S435/911—Microorganisms using fungi
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
    • Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10S435/00—Chemistry: molecular biology and microbiology
    • Y10S435/8215—Microorganisms
    • Y10S435/911—Microorganisms using fungi
    • Y10S435/921—Candida
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
    • Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10S435/00—Chemistry: molecular biology and microbiology
    • Y10S435/8215—Microorganisms
    • Y10S435/911—Microorganisms using fungi
    • Y10S435/93—Hansenula
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
    • Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10S435/00—Chemistry: molecular biology and microbiology
    • Y10S435/8215—Microorganisms
    • Y10S435/911—Microorganisms using fungi
    • Y10S435/938—Pichia

Definitions

  • Tanenholtz Attorney Maisroth, Lind & Ponack ABSTRACT An optically active compound having a 13,8-carbon-substituted-l 7-hydroxy-8, l 4-secogona- 1,3,5( l0),9( l l ),l5-pentaen-l4-one nucleus is produced from a compound having a l3B-carbon-substituted-8,l4-secogona- 1,3,5(10),9(1 l),l5-pentaene-14,17-dione nucleus by the action on the latter of the redox-enzyme system obtained by culturing a Candida, Debaryomyces, Kloeckera, Pichia, Rhodotorula, Schizosaccharomyces or Hansenula micro-organism.
  • FIG 1F ZOGKMEQQ m kOm 350 WAVE LENGTH (mu)
  • FIGJE O ZOEKMEQQ m kOm INVENTORS MASAO ISONO M HA 2 A K AMM T IY H mm EHU 7 MYT Y B L m5 m ATTORNEYS PATENTEDum 26 um
  • SHEET 30F 3 24 HOURS
  • FIGZB WAVELENGTH w)
  • the present invention relates to a process for producing optically active unsaturated steroids.
  • the present invention relates to a process 5 for producing an optically active compound having a l3fl-carbon-substituted-l 7-hydroxy-8 ,14-secogona-polyenl 4-one nucleus [hereinafter referred to as compound (")1 from a compound having a l3fi-carbon-substituted-8,14-secogona- 1,3,5( 10), 9(l l),lS-pentaenel 4,1 7 dione nucleus [hereinafter referred to as compound (l)] through the utilization of a micro-organism.
  • compound (l) an optically active compound having a l3fl-carbon-substituted-l 7-hydroxy-8 ,14-secogona-polyenl 4-one nucleus
  • the compounds attainable by those methods are invariably 50:50 mixtures of a compound having a lElfi-carbon-substituted-l7a-hydroxy-8, l 4-secogonal,3,5(l0),9( l l) -tetraen-l4-one nucleus and its enantiomer, and in order to convert them into natural estrone and the like, it is necessary to subject them to an optical resolution at some stage.
  • the main object of the present invention is to provide a novel and industrially feasible process for producing compound (ll) directly from compound (I) in a good yield.
  • the object ofthe present invention is realized by incubating a micro-organism belonging to the genus Candida, Debaryomyces, Kloeckera, Pichia, Rhodotorula, Schizosaccharomyces or Hansenula, contacting the redox-enzyme system of the thus-obtained culture with a compound having a l3,B-carbonsubstituted-8,l4-secogona-1,3,5( l0),9(] l),l5-pentaenel4,l 7-dione nucleus, and recovering the objective compound having a l3fl-carbon-substituted-l7-hydroxy-8,14-secogonapolyenl 4-one nucleus from the reaction mixture.
  • substituents at the l3-carbon there may be exemplified lower alkyl, such as methyl, ethyl, propyl, butyl; aryl, such as phenyl; aralkyl such as benzyl, phenethyl, phenyl propyl, etc.
  • the compound (I) employed as the starting material which can be produced, for example, by the method described in the Netherlands Pat. application No. 6,612,205 (published on Mar. 1, 1967), has a substituent at the l3-carbon and one to three other substituents-oxygen groups, halogen atoms and lower alkyl groups-at any of the l-,2-,3-,4-,6-,7-,1l-,l2-,l5- and lo-positions.
  • the oxygen groups include, for example, the hydroxy group, a lower acyloxy group containing one to four carbon atoms, an alkoxy group having one to four carbon atoms, and the benzyloxy group while the halogens include fluorine, chlorine and bromine, for instance.
  • the lower alkyl groups are straight chain of one to four carbon atoms.
  • the objective compounds (II) can be classified as follows:
  • the nutritive media for the growth of such micro-organisms contain the sources of carbon and nitrogen which can be utilized by the said micro-organisms and they may contain inorganic salts, various vitamins, and amino acids.
  • the carbon sources include, for example, glucose, sucrose, dextrin, glycerol and others and the nitrogen sources include, for example, such organic nitrogenous materials as peptone, meat extract, casein, corn steep liquor, dry yeast and yeast extract and such inorganic nitrogen compounds as ammonium nitrate, ammonium phosphate, ammonium sulfate, and the like.
  • the above-mentioned inorganic salts include sodium chloride, potassium sulfate, magnesium sulfate and the like. Those nutrients promoting the growth of micro-organisms are employed in suitable proportions to make up a culture medium.
  • the cultivation may be carried out by any of the shake culture, the stationary culture and the submerged culture methods with agitation and aeration.
  • Addition of starting compound (I) can be made either at the start of the cultivation or at a suitable stage in the course of the cultivation, continuously or at some intervals or at one stroke, the final concentration of the compound (l) in the medium being advantageously between about 0.05 percent and about 0.6 percent (weight by volume basis).
  • the compound (I) can be employed in the form of a powder or as a solution or suspension in a suitable solvent, which may be acetone, methanol, ethanol, ethylene glycol, propylene glycol, dimethylformamide or dioxane, for instance, with or without addition of a surface active agent, dispersing agent and the like.
  • a suitable solvent which may be acetone, methanol, ethanol, ethylene glycol, propylene glycol, dimethylformamide or dioxane, for instance, with or without addition of a surface active agent, dispersing agent and the like.
  • the; microbial cells can be suspended in a buffer solution of suitable pH and ionic strength, or in water, and the starting compound (I) contacted therewith so as to convert the latter to compound (ll).
  • reaction proceeds at a pH from about 2 to about 11, and at a temperature somewhere between about 10 and about 50 C., advantageously between about 25 and about 40 C. for about 2 days.
  • the optimum conditions vary with such factors as the starting compounds and the micro-organisms and the optimum conditions are selected in each particular situation.
  • the final compound (ll) thus produced and accumulated in the reaction medium can be isolated by various per se known means.
  • such separation means are available as adsorption by which the end product is adsorbed on a suitable adsorbent, e.g. silica gel, followed by elution with a suitable solvent such as benzeneacetone mixture or chloroformacetone mixture the means utilizing the difference in distribution coefficient between two liquid phases, for example, counter current distribution techniques, and conventional chromatographic techniques.
  • a suitable adsorbent e.g. silica gel
  • optically active compounds with a l3B-carbon-substituted-l7-hydroxy-8,l4-secogona-1,3,5( l),9( l l ),l5-pentaen-l4-one nucleus are converted to the corresponding l,3,5( l0),bh9( l l) -tetraenes by the use of the enzyme system of a yeast, for example, Candida solani, after the manner of the process of the present invention.
  • the optically active compounds having a 13B-carbon-substitutedl 7-hydroxy-8, l 4-secogona-l ,3,5( l0),9(l l) tetraen-l4-one nucleus prepared above, the compounds having a l 3fl-carbon-substituedl 7a-hydroxy-8, l4-secogonatetraen-l4-one nucleus are converted to estrone or other useful l9-nor steroids for example, by the method described in French Pat. No. 1,526,031.
  • the compounds having a 13B-carbonsubstitutedl 7B-hydroxy-8, l 4-secogona-tetraenl 4-one nucleus are, for example, firstly subjected to cyclization in benzene with p-toluene sulfonic acid to give the optically active compounds having a 13B-carbon-substituted-l7B-hydroxy-gona-l,3,5(lO),8,l4-pentaene nucleus, which are then led to estrone through a series of the reactions disclosed in British Pat. No. 1,064,01 l.
  • part(s) by weight and part(s) by volume is the same as that between gram(s) and milliliter(s).
  • yeast extract 0.5 percent of polypeptone, 0.2 percent of corn steep liquor, 5 percent of glucose and 5 per cent of sucrose are inoculated respectively with the 7-day slant culture of a micro-organism shown in table 1.
  • the inoculated medium is incubated at 28 C. for 2 days, at the end of which period there is added thereto a solution of 0.02 part by weight of 3-mcthoxy-8,l4-secoestra-1,3,5( l0),9( l l ),l5-pentaene-l4,l7-dione [hereinafter referred to as substrate (l)] in 0.8 part by volume of ethanol.
  • substrate (l) 3-mcthoxy-8,l4-secoestra-1,3,5( l0),9( l l ),l5-pentaene-l4,l7-dione
  • the resulting broth is mixed twice together with an equal amount of ethyl acetate, and the reaction products transferred to the ethyl acetate phase are washed with water, dehydrated and concentrated to dryness.
  • EXAMPLE 3 1n the same manner as in example 2, a solution of one part by weight of substrate (1) in 40 parts by volume of ethanol is added to 1,000 parts by volume of a 2-day culture of Pichia elchellsii UFO-1283; ATCC 20126) which is then incubated for further 2 days.
  • EXAMPLE 4 In the same manner as in example 2, eight parts by volume of 2.5 percent solution of substrate (I) in ethanol is added to 200 parts by volume of a 2-day culture of Hansenula holstii 1FO-0980 which is then incubated at 28 C. The sampling is made at 12, 24, 36 and 48hours after addition of the substrate (1), and the samples are respectively subjected to extraction with ethyl acetate. The extracts are purified by column chromatography on silica'gel and are dissolved in ethanol.
  • the product attained after 36 and 418 hours of cultivation, respectively, is crystallized from ethanol in the form of colorless plates melting at 111 to 113 C. (Yield 78 percent on weight basis).
  • the [01],, and infrared absorption spectra of the products are in perfect agreement with those of 3-methoxy-8,14-secoestra-1,3,5(10),9(11)-tetraen-17/3-o1-l4-one.
  • Hansenula holstii IFO-0980 has the property to first reduce the 17-carbonyl group of substrate (1) to yield 3-methoxy-8,l4-secoestra-l ,3,5 (l0),9(l 1),l5-pentaen-17B-ol-14-one and then reduce the A of the latter to yield 3-methoxy-8,14-secoestra-l,3,5 (10),9(1l)-tetraen-l7A-ol-14-one.
  • EXAMPLE 5 In the same manner as in example 2, a solution of 0.5 part by weight of substrate (1) in 40 parts by volume of ethanol is added to 1000 parts by volume of a 2day culture of Debaryomyces vanriji (IFO-l285; ATCC 20125) which is then cultivated as such for 2 days. The reaction product is extracted with ethyl acetate and the extract is purified by column chromatography after the manner described in example 2 to give 0.35 part of 3-methoxy-8,l4-secogona-l,3,5(10),9(l1),l5- pentaen-l 7B-ol-l4-one as an oil.
  • EXAMPLE 6 In the same manner as in example 2, 40 parts by volume ofa 2.5 percent solution of substrate (1) in. ethanol is added to 1,000 parts by volume of a 2-day culture of Kloeckera magna UFO-0868; ATCC 20131) and the reaction is allowed to proceed at 28 C. for 2 days, at the end of which period the product is extracted with ethyl acetate.
  • the extract is purified by column chromatography with silica gel after the manner described in example 2 to give 3- methoxy-8,l4-secoestra-l,3,5( l0),9,5-pentaen-l 7a-oll 4- one as an oil (yield 75 percent on weight basis),
  • the nuclear magnetic resonance spectrum of the productin CDC]; (Dzdeuterium) displays characteristic peaks of the A- l7-ol structure: pl l7-H (r 5.52, doublet), 16-l-l(r 2.62 quartet) and 15-1-1 ('r 3.89 quartet).
  • a process for producing an optically active compound having a l3B-carbon-substitutedl 7-hydroxy-8,l4-secogonapolyen-l4-one nucleus which comprises subjecting a compound having a 13/3-carbon-substituted-8,l4-secogonal,3,5(l0),9(1l),l5-pentaene-l4,l7-dione nucleus to the action of an enzyme system of Candida utilis, Debaryomyces vini, Debaryomyces vanriji, Debaryomyces nicotianae, Kloeckera magna, Pichia wickerhamii, Pichia pijperi, Pichia etchellsii, Rhaa'otorula rubra, Hansenula capsulala, Hansenula holslii or Hansenula saturnus', and recovering the objective product from the reaction mixture.

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  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
US746882A 1967-07-24 1968-07-23 Process for producing unsaturated steroids Expired - Lifetime US3616226A (en)

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US (1) US3616226A (enrdf_load_stackoverflow)
BE (1) BE718490A (enrdf_load_stackoverflow)
DE (1) DE1768985C3 (enrdf_load_stackoverflow)
FR (1) FR1581108A (enrdf_load_stackoverflow)
GB (1) GB1239602A (enrdf_load_stackoverflow)
NL (1) NL158152B (enrdf_load_stackoverflow)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3793148A (en) * 1972-07-24 1974-02-19 Syntex Corp Microbiological reduction of 1,3,-dioxo-2-alkylcycloalkanes
US5773264A (en) * 1992-03-28 1998-06-30 Schering Aktiengesellschaft Process for the production of 17 α-hydroxy-3-methoxy-8,14-seco-1,3,5(10),9(11)estratetraen-14-one by reduction of the corresponding 17-one compound
US20110020887A1 (en) * 2006-12-07 2011-01-27 Iep Gmbh Process for the enantioselective enzymatic reduction of secodione derivatives

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3697379A (en) * 1970-04-28 1972-10-10 American Home Prod Asymmetric reduction of seco-steroids

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3793148A (en) * 1972-07-24 1974-02-19 Syntex Corp Microbiological reduction of 1,3,-dioxo-2-alkylcycloalkanes
US5773264A (en) * 1992-03-28 1998-06-30 Schering Aktiengesellschaft Process for the production of 17 α-hydroxy-3-methoxy-8,14-seco-1,3,5(10),9(11)estratetraen-14-one by reduction of the corresponding 17-one compound
US20110020887A1 (en) * 2006-12-07 2011-01-27 Iep Gmbh Process for the enantioselective enzymatic reduction of secodione derivatives
EP2410047A1 (de) 2006-12-07 2012-01-25 IEP GmbH Oxidoreduktase und deren Verwendung zur Reduktion von Secodionderivaten
US8323936B2 (en) 2006-12-07 2012-12-04 Iep Gmbh Process for the enantioselective enzymatic reduction of secodione derivatives

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DE1768985C3 (de) 1980-12-18
DE1768985A1 (de) 1972-01-05
DE1768985B2 (de) 1980-04-17
FR1581108A (enrdf_load_stackoverflow) 1969-09-12
NL158152B (nl) 1978-10-16
GB1239602A (enrdf_load_stackoverflow) 1971-07-21
NL6810400A (enrdf_load_stackoverflow) 1969-01-28
BE718490A (enrdf_load_stackoverflow) 1968-12-31

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