CH620459A5 - Process for the preparation of D-homosteroids - Google Patents

Abstract

D-Homosteroids of the formula <IMAGE> in which R<6> is hydrogen, fluorine, chlorine or methyl; R<17a> is hydroxyl or acyloxy and R<20> is hydrogen, lower alkyl, halo-lower-alkyl, hydroxy-lower-alkyl, acyloxy-lower-alkyl or lower alkoxycarbonyl-lower-alkyl, and the dotted 1,2 bond is an optional C-C bond, are prepared by 11-hydroxylation of a corresponding D-homosteroid which is unsubstituted in position 11 using microorganisms or enzymes obtained therefrom. The compounds have an endocrine, in particular corticoid-like, action.

Description

620459

PATENT CLAIMS 1. Process for the preparation of D-homosteroids of the formula

COOR20

delt and a 6ß-substituted D-homosteroid of formula I obtained isomerized to the 6a isomer by treatment with an acid.

7. A process for the preparation of a 17a-acylated D-homo-steroid of the formula I, characterized in that a 17a-hydroxy-D-homostere-roid of the formula I is prepared by the process of claim 1 and then the 17a-hydroxy group is acylated .

8. The method according to claim 1, characterized in that 1,2-unsaturated D-homosteroids of the formula I are prepared.

9. The method according to claim 1, characterized in that 17aa-acetoxy-l l-hydroxy-3-oxo-D-homoandrosta-l, 4-diene-17 aß-carboxylic acid methyl ester is prepared.

in the R6 hydrogen, fluorine, chlorine or methyl; R17a Hydroxy 20 The invention relates to a process for the preparation of or acyloxy and R20 hydrogen, lower alkyl, halo-lower-new D-homosteroids of the formula alkyl, hydroxy-lower alkyl, acyloxy-lower alkyl or lower alkoxycarbonyl-lower alkyl and the dotted 1,2 Bond mean an optional CC bond, characterized in that

COOR20

that you have a D homosteroid of the formula

--R

II

hydroxylated in the 11-position by means of microorganisms or enzymes obtained therefrom.

2. A process for the preparation of a compound of the formula I in which R20 is different from hydrogen, characterized in that a compound of the formula I in which R20 is hydrogen is prepared by the process of claim 1, and this with a group other than hydrogen R20 donating agent treated.

3. A process for the preparation of an A4-D homosteroid of the formula I, characterized in that an A '^ - D-homosteroid of the formula I is prepared by the process of claim 1 and the 1,2-double bond is then hydrogenated.

4. A process for the preparation of an A1,4-D-homosteroid of the formula I, characterized in that a 1,2-saturated D-homosteroid of the formula I is prepared by the process of claim 1 and this is then in the 1,2-position dehydrated.

5. A process for the preparation of a D-homosteroid of the formula I in which R6 is fluorine or chlorine, characterized in that a compound of the formula I in which R6 is hydrogen is prepared by the process of claim 1, and this with a fluorination or Chlorinating agents treated.

6. A process for the preparation of a D-homosteroid of the formula I in which R6 is fluorine or chlorine, characterized in that a compound of the formula I in which R6 is hydrogen is prepared by the process of claim 1, and this with a fluorination or Chlorinating agents treated

17a

65

in the R6 hydrogen, fluorine, chlorine or methyl; R17a is hydroxy or acyloxy and R20 is hydrogen, lower alkyl, halo-lower alkyl, hydroxy lower alkyl, acyloxy-lower alkyl or lower alkoxycarbonyl-lower alkyl and the dotted 1,2 bond means an optional C-C bond.

An acyloxy group can be derived from a saturated or unsaturated aliphatic carboxylic acid, a cycloaliphatic, araliphatic or an aromatic carboxylic acid with preferably up to 15 C atoms. Examples of such acids are: formic acid, acetic acid, pivalic acid, propionic acid, butyric acid, caproic acid, oenanthic acid, undecylenic acid, oleic acid, cyclopentylpropionic acid, cyclohexylpropionic acid, phenylacetic acid and benzoic acid. Ci_7-alkanoyloxy groups are particularly preferred. Lower alkyl groups can be straight or branched chain and contain 1 to 6 carbon atoms. Lower alkyl groups with 1-4 carbon atoms, in particular methyl and ethyl, are particularly preferred. The terms halo-lower alkyl, hydroxy-lower alkyl, acyloxy-lower alkyl and lower alkoxy-carbonyl-lower alkyl are to be understood in the same way. The term “halo” includes, unless specifically defined, fluorine, chlorine, bromine and iodine. Examples of halo-lower alkyl radicals are fluoromethyl, chloromethyl, bromomethyl, ß-fluoroethyl, ß-chloroethyl and ß-bromoethyl. Examples of hydroxy (and acyloxy) lower alkyl are ß-hydroxyethyl and ß-acetoxy-ethyl. A lower alkoxycarbonyl-lower alkyl radical is, for example, methoxycarbonylmethyl.

A preferred group of compounds of the formula I are the compounds of the formula I with a 1,2-double bond.

3rd

620 459

Examples of compounds obtainable according to the invention are:

11 ß, l 7 aa-dihydroxy-3-oxo-D-homoandrost-4-en-17 aß-carboxylic acid,

llß, 17aa-dihydroxy-3-oxo-D-homoandrosta-l, 4-diene-17ass-car-5 acid,

6a-ChIor-l 1 ß, l 7 aa-dihydroxy-3-oxo-D-homoandrost4-en-l 7aß-carboxylic acid,

11β, 17a-dihydroxy-6a-methyl-3-oxo-D-homoandrost-4-en-17ass-carboxylic acid, io the methyl, ethyl, propyl and butyl esters of these compounds and the 17aa acetates, - Propionates and butyrates.

According to the invention, the D-homosteroids of the formula I are prepared by using a D-homosteroid of the formula 15

II

hydroxylated in the 11-position by means of microorganisms or enzymes obtained therefrom.

The 1,2-dehydrogenation of a D-homosteroid of the formula I can be carried out in a manner known per se, e.g. B. microbiological 35 ways or by means of dehydrating agents such as iodine pentoxide, periodic acid or selenium dioxide, 2,3-dichloro-5,6-dicyanobenzo-quinone, chloranil or lead tetraacetate. Suitable microorganisms for 1,2-dehydrogenation are, for example, schizomycetes, in particular those of the Genera 40 Arthrobacter, e.g. B. A. simplex ATCC 6946; Bacillus, e.g. B. B. len-tus ATCC 13805 and B. sphaericus ATCC 7055; Pseudomonas, e.g. B. P. aeruginosa IFO 3505; Flavobacterium, e.g. B. F. flavens-cens IFO 3058; Lactobacillus, e.g. B. L. brevis IFO 3345 and Nocardia, e.g. B. N. opaca ATCC 4276. 45

The halogenation of a steroid of formula I in 6th position (process variant c) can be carried out in a manner known per se. A 6,7-saturated D-homosteroid of the formula I can be halogenated by reaction with a halogenating agent, such as an N-chloramide or imide (e.g. N-chlorosuccin 50 imide) or with elemental chlorine [cf. J. Am. Chem. 72, 4534 (1950)]. The halogenation in the 6-position is preferably carried out by adding a 6,7-saturated D-homosteroid of the formula I into a 3-enol ester or 3-enol ether, e.g. B. the 3-enol acetate, and then 55 with chlorine [cf. J. Am. Chem. Soc. 82, 1230 (I960)]; with an N-chloroimide [cf. J. Am. Chem. Soc. 82, 1230 (1960); 77, 3827 (1955)] or perchloryl fluoride [cf. J. Am. Chem. Soc. 81, 5259 (1959); Chem. And Ind. 1959, 1317]. Trifluoromethyl hypofluorite is also considered as a fluorinating agent in 6o.

If isomer mixtures in the halogenations described above, i. H. Mixtures of 6a and 6β-halogen steroids are formed, these can be separated into the pure isomers by known methods, such as chromatography.

The isomerization of a 6β-halogen-D-homo-steroid obtained, i.e. H. a 6ß-fluorine or chlorine compound can by

Treatment with an acid, especially a mineral acid, such as hydrochloric acid, or hydrobromic acid in a solvent, e.g. B. dioxane or glacial acetic acid.

The hydroxylation of a compound of formula II according to the invention can be accomplished by means of methods known per se for the microbial 11-hydroxylation of steroids. Microorganisms of the taxonomic unit Fungi and Schizomycetes, in particular of the sub-units Ascomycetes, Phycomycetes, Basidiomycetes and Actinomycetales, are suitable for this. It can also by chemical means, e.g. B. by treatment with nitrite, or by physical means, e.g. B. by radiation, mutants generated and cell-free enzyme preparations obtained from the microorganisms can be used. Suitable microorganisms for 1β-hydroxylation are, in particular, those of the Genera Cur-vularia, e.g. B. C. lunata NRRL 2380 and NRRL 2178; ATCC 13633.13432.14678, IMI 77007, IFO 2811; Absidia, e.g. B. A. coe-rula IFO 4435; Colletotrichum, e.g. B. C. pisi ATCC 12520; Pelli-colaria, e.g. B. P. filamentosa IFO 6675; Streptomyces, e.g. B. S. fra-diae ATCC 10745; Cunninghamella, e.g. B. C. bainieri ATCC 9244, C. verticellata ATCC 8983, C. elegans NRRL 1392 and ATCC 9245, C. blakesleeana ATCC 8688,8688a, 8688b, 8983, and C. echinulata ATCC 8984; Pycnosporium, e.g. E.g. ATCC 12231; Verticillium, e.g. B. V. theobromae CBS 39858; Aspergillus, e.g. B. A. quadrilieatus JAM 2763; Trichothecium, e.g. B. T. roseum ATCC 12519; and Phoma, e.g. E.g. ATCC 13145.

The acylation of a 17a-hydroxy group can be carried out in a manner known per se, e.g. B. by treatment with an acylation agent such as an acyl chloride or anhydride in the presence of an acid binding agent, e.g. B. pyridine or triethylamine or in the presence of a strong acid catalyst, e.g. B. p-toluenesulfonic acid. Non-hydroxyl organic solvents, e.g. B. chlorinated hydrocarbons such as methylene chloride or hydrocarbons such as benzene. It is also possible to first convert a 17aa-hydroxy-17ass-carboxylic acid D-homosteroid of the formula I with a corresponding carboxylic acid anhydride into a mixed anhydride of the steroid carboxylic acid and to convert this mixed anhydride into acidic or basic (e.g. with aqueous acetic acid or aqueous pyridine), whereby the desired 17aa-acyloxy derivative of the starting compound I is obtained.

The functional modification of the group -COOR20 z. B. in an esterification of a 20-carboxyl group, a transesterification of an esterified 20-carboxyl group or an esterification of a hydroxyl group contained in the radical R20. All of these reactions can be carried out according to methods known per se. The esterification can e.g. B. by treating the free acid with a diazoalkane, e.g. B. diazomethane in ether; or with an 0-alkyl-N, N'-dicyclohexylisourea in an aprotic solvent; or by reacting a salt of the acid, e.g. An alkali salt with an alkyl halide or sulfate, e.g. As methyl or ethyl iodide or dimethyl or diethyl sulfate can be achieved.

The acylation of a hydroxyl group contained in the radical R20 can be accomplished analogously to the acylation of a 17aa-hydroxyl group described above. The transesterification of an esterified carboxyl group, e.g. B. The exchange of an alkyl group represented by R20 for another alkyl group can be carried out by reaction with the corresponding alcohol in the presence of an acidic catalyst such as perchloric acid. Compounds in which R20 represents a halogen- or hydroxy-substituted alkyl group can e.g. B. be prepared by reacting a salt of a D-homosteroid carboxylic acid of the formula XII with a sulfonyloxyalkyl halide to give a sulfonyloxyalkyl ester and hydrolyzing the latter to give a hydroxyal-

620 459 4

to obtain alkyl substituents R20; or with an alkali or total substrate use of 22.8 g alkaline earth halide, z. B. lithium chloride in dimethylformamide, and by column chromatography on silica gel with a trench to obtain a haloalkyl substituent R20, elution of methylene chloride: methylene chloride / acetone haloalkyl esters can also be obtained by (3/1) 1: 1 purification. After recrystallization from vinegar the D-homosteroid carboxylic acid with an appropriate 5 ester 8.1 g 17 aa-acetoxy-11β-hydroxy-3-oxo-D-homo-androst-the aldehyde in the presence of hydrogen halide, 4 -en-l 7ass-carboxylic acid methyl ester (34.3% of theory) melting point moderately in the presence of a catalyst such as ZnCh. 239 / 240-241 °.

The hydrogenation of a 1,2-double bond in an analogous manner gave the 11β, l 7aa-dihydroxy-3-

D-homosteroid of formula I can be catalytic, e.g. B. with Pd or oxo-D-homoandrosta-1,4-diene-l 7ass carboxylic acid from mp. Tris (triphenylphosphine> rhodium chloride 10 237-238 ° C; UV: £ 243 = 14600; die6a-fluorine -llß, 17aa-dihy-den. droxy-3-oxo-D-homoandrosta-l, 4-dien-17aß-carboxylic acid; UV:

The raw materials, if not known, or 82 «= 14 900.

are described below, in analogy to known methods or described in the examples. Example 2

The D-homosteroids of the formula I are endocrine, in particular 15 A solution of 120 mg of 11β, l 7aa-dihydroxy-3-oxo-D-other anti-inflammatory activity. They have a good homoandrost-4-en-l 7ass carboxylic acid and 0.13 ml triethylamine ratio of the anti-inflammatory effect to effects in 3.1 ml CH2CI2 was added dropwise at 0 ° C with 0.88 ml Ace mineral or glucocorticoider Nature on. tyl chloride added and stirred for 40 minutes. The reaction

The process products can be mixed as a remedy in the form of methylene chloride and successively pharmaceutical preparations with direct or delayed 20 with 3% sodium hydrogen carbonate solution, IN hydrochloric acid and release of the active ingredient in a mixture with one for enteral, water washed, dried and evaporated. The back percutaneous or parenteral application suitable organism was dissolved in 3.1 ml acetone and see with 0.1 ml diethylamine or inorganic inert carrier material, such as. B. offset. The mixture was concentrated in vacuo, the water, gelatin, gum arabic, milk sugar, starch, with the precipitate which had precipitated, filtered off, dissolved in water,

Magnesium stearate, talc, vegetable oils, polyalkylene glycols, 25 acidified with 2N hydrochloric acid and extracted with ethyl acetate. Vaseline, etc. can be used. The pharmaceutical pre- After drying and evaporation of the solvent was ready parate in solid form, e.g. B. as tablets, dragees, sup-17aa-acetoxy-11ß-hydroxy-3-oxo-D-homoandrost-4-en-17aß-positorien, capsules; in semi-solid form, e.g. B. as ointments; or in carboxylic acid of mp. 167-170 ° C (from acetone / hexane) containing liquid form, for. B. as solutions, suspensions or emulsions; UV: e24i = 13,900; [a] D = + 39 ° (c = 0.1% in dioxane). NEN. If necessary, they are sterilized and / or 30 In an analogous manner it was produced: from 11β, l 7aa-dihy or contain auxiliaries such as preservative, stabilizing-hydroxy-3-oxo-D-homoandrost-4-en-17 ate -carbonsäuremethyl-

Rungs-, wetting or emulsifying agents, salts for changing the ester, the 17 aa-acetoxy-1 lß-hydroxy-3-oxo-D-homoandrost-4-osmotic pressure or buffer substances. en-17ass-carboxylic acid methyl ester, mp. 235-237 ° C (from ether);

As a guideline for the composition of topical prepa- UV: e242 = 15 750; [a] D = + 49 ° (c = 0.1% in dioxane); from a rate of about 0.01-1% content of a compound of the formula 35 1 lß, l 7aa-dihydroxy-3-oxo-D-homoandrosta-l, 4-dien-17aß-car-mel I. In the case of systemic preparations, about 0.1-10 bonic acid, the 17aa-acetoxy-1-beta-hydroxy-3-oxo-D-homoandro-mg active ingredient can be provided per application. sta-l, 4-dien-17aß-carboxylic acid of mp 251 ° C (dec.), UV: e243

The preparation of the pharmaceuticals can be done in a manner known per se = 14,500; [a] D = + 25 ° (c = 0.1% in methanol); from a 6a-fluorine manner by adding the compounds of the formula I with 1ß, 17aa-dihydroxy-3-oxo-D-homoandrosta-1, 4-diene-17ass-car-suitable for therapeutic administration, non-toxic 40 bonic acid, the 17aa-acetoxy-6a-fluoro-l 1 ß-hydroxy-3-oxo-D's, solid and / or homoandrosta-l, 4-diene-l 7ass-carboxylic acid, customary in such preparations, of mp 208 ° C (from liquid carrier materials, such as the above-mentioned acetone / hexane), UV: e243 = 14,650; [a] D = + 21 ° (c = 0.1% in ten, mixed and optionally in the desired form methanol).

brings.

45 Example 3

Example 1 50 mg 11 β, 1 7 aa-dihydroxy-3-oxo-D-homoandrost-4-en-

A 2-1 Erlenmeyer flask containing 500 ml of a 30 minutes at 17ass carboxylic acid was dissolved in 3 ml of methanol and at 0 ° 120 ° sterilized in an autoclave nutrient solution from 3.0% glucose, with 2.6 ml of 0.06N ethereal diazomethane solution transferred. 1.0% Cornsteep, 0.2% NaNOî, 0.1% KH2PO4.0.2% K2HPO4, after 5 minutes a few drops of acetic acid were added containing 0.05% MgS04.0.002% Fe SO4 and 0.04% KCl, is evaporated at 50 and the mixture. Filtration in 1 g alumina of a lyophil culture from Trichothecium roseum (ATCC 12519) gave 1 lß, 17aa-dihydroxy-3-oxo-D-homoandrost-4-en-17ass-car-inoculated and 72 hours at 30 ° on a rotary shaker of methyl bonsate M.p. 186-188 ° C (from ether), UV: shaken. This preculture is then used to make a 20-1 fermenter e243 = 15 750, [a] D = + 95 ° (c = 0.1% in dioxane).

made of stainless steel, which inoculates with 151 of the sterile at 121 ° and 1.1 atm. In an analogous manner, was prepared: from 17aa-acetoxy-containing medium of the same composition. 55 11 ß-hydroxy-3-oxo-D-homoandrost-4-en-l 7ass-carboxylic acid, the addition of silicone SH as an anti-foaming agent in 17aa-acetoxy-l 1 ß-hydroxy-3-oxo-D-homoandrost -4-en-l 7aß-29 ° C with aeration (101 / min.) 0.7 atm pressure and stirring (220 carboxylic acid methyl ester, mp. 235-237 ° C (from ether), UV: U / min.) 24 Hours cleaned, 1.81 of the culture broth is under 15 750, [a] D = + 49 ° (c = 0.1% in dioxane); from 17aa-acetoxy-l Iß-sterile conditions in 251 of a medi- hydroxy-3-oxo-D-homoandrosta-l, 4-diene-17ass-carboxylic acid sterilized as above, which is converted to the same composition and under the same bo 17aa-acetoxy -l lß-hydroxy-3-oxo-D-homoandrosta-l, 4-diene conditions. After 12 hours, a sterile-17ass-carboxylic acid methyl ester of mp 235-237 ° C., UV: e242 = triated solution of 6.6 g of 17aa-acetoxy-3-oxo-D-homo-androst-15 100, [a] D = + 13 ° (c = 0.1% in dioxane); from 17aa-acetoxy-6a-4-en-17ass-carboxylic acid methyl ester in 200 ml of dimethylform-fluor-1-lß-hydroxy-3-oxo-D-homoandrosta-l, 4-diene-17ass-carbon-amide. After a contact time of 19 hours, the ferrous acid, the 17aa-acetoxy-6a-fluoro-1 1 ß-hydroxy-3-oxo-D-homo-menthol content is extracted twice with 151 methyl isobutyl ketone each 65 androsta-l, 4-diene 17ass-carboxylic acid methyl ester of mp.

stirred and the extract at 50 ° bath temperature in a vacuum 238-239 ° C, UV: e242 = 15 550, [<x] D = + 31 ° (c = 0.1% in metha vapor) The residue is removed silicone oil noi); washed several times with hexane from Hβ-hydroxy-3-oxo-17aa-propionyloxy-D-homoand. Several approaches with a rust-4-en-l 7ass-carboxylic acid, the

11 ß-hydroxy-3-oxo-l 7 aa-propionyloxy-D-homoandrost-4-en-l 7-aß-carboxylic acid methyl ester, UV: e24i = 14 900, [a] D = + 48 ° (c = 0.102% in dioxane).

Example 4

A solution of 3.2 g of 1 liter, 17aa-dihydroxy-3-oxo-D-homo-androst-4-en-17ass-carboxylic acid and 4.2 ml of triethylamine in 86 ml of CH2Cl2 was added dropwise at 0 ° under argon with 29 ml of propionyl chloride were added and the mixture was stirred for 45 minutes. The mixture was poured onto ice cold dilute hydrochloric acid and extracted three times with methylene chloride. The organic solution was washed with brine, dried and evaporated. Any remaining propionic acid was removed by repeated evaporation with toluene. The 11β-hydroxy-3-oxo-l 7 aa-propionyloxy-D-homoandrost-4-en-17ass-carboxylic acid was obtained as a non-crystalline foam. UV: e24i = 15,000, [a] D = + 37 ° (c = 0.1% in dioxane).

Example 5

420 mg of 11β-hydroxy-3-oxo-17aa-propionyloxy-D-homo-androst-4-en-17ass-carboxylic acid, 0.5 ml of triethylamine and 0.5 ml of n-butyl iodide were refluxed in 10 ml of acetone for 48 hours - river cooked. The reaction mixture was poured onto ice-cold, dilute hydrochloric acid and extracted with CH2Cl2. The organic solution was washed with brine, dried and evaporated to give butyl 1β-hydroxy-3-oxo-17aa-propionyloxy-D-homoandrost-4-ene-17ass-carboxylic acid, mp 150-151 ° C (from acetone / hexane ) was obtained. UV: £ 24i = 16 150, [a] o = + 41 ° (c = 0.104% in dioxane).

In an analogous manner, 17aa-acetoxy-lß-hydroxy-3-oxo-D-homoandrosta-l, 4-diene-17ass-carboxylic acid and Methyljo-did, the 17aa-acetoxy-l lß-hydroxy-3-oxo D-homoandrosta-l, 4-diene-17ass-carboxylic acid methyl ester (identical to the compound obtained in Example 3).

Example 6

900 mg of 1 lß-hydroxy-17aa-propionyloxy-3-oxo-D-homo-androst-4-en-l 7ass-carboxylic acid were added in 3 ml of DMF with 0.5 ml of triethylamine and 1 ml of chlorine iodomethane and stirred at room temperature . ÌAfter 24 and 48 hours, another 0.5 ml of triethylamine and 1 ml of chloroiodomethane were added. After a total of 72 hours, the mixture was poured onto ice-cold, dilute hydrochloric acid, extracted with CH2Cl2, washed with dilute sodium chloride solution, dried and evaporated. Chromatography on silica gel gave 11β-hydroxy-17aa-propionyloxy-3-oxo-D-homoandrost-4-en-l 7 aß-carboxylic acid, chloromethyl ester of mp 180-181 ° C, UV: 6242 = 17350, [a] D = + 82 ° (c = 0.104% in dioxane).

Example 7

500 mg of 1 l-hydroxy-17aa-propionyloxy-3-oxo-D-homo-androst-4-en-17ass-carboxylic acid, chloromethyl ester in 1 ml of silver fluoride were added to 65 ml of acetonitrile and stirred. After 3 days, another 0.5 g of silver fluoride was added. After a further 3 days, the reaction mixture was diluted with ethyl acetate and filtered through silica gel. The eluate was washed with water, dried and evaporated. Chromatography on silica gel gave 1 lß-hydroxy-17aa-propionyloxy-3-oxo-D-homoandrost-4-en-17ass-carboxylic acid fluoromethyl ester of mp 171 ° C, UV: e240 = 23,700, [a] D = + 59 ° (c = 0.101% in dioxane).

620459

Example 8

360 mg of 17aa-acetoxy-1-beta-hydroxy-3-oxo-D-homoandrost-4-en-17ass-carboxylic acid methyl ester and 250 mg of selenium dioxide were refluxed in 20 ml of t-butanol and 0.2 ml of glacial acetic acid under argon for 20 hours . The mixture was filtered and evaporated. The oil obtained was dissolved in ethyl acetate and washed successively with sodium hydrogen carbonate solution, water, ice-cold ammonium sulfide solution, dilute ammonia, water, dilute hydrochloric acid and water. The ethyl acetate solution was dried over sodium sulfate and evaporated. Chromatography on silica gel gave 17aa-acetoxy-11β-hydroxy-3-oxo-D-homoandrosta-l, 4-diene-l 7 aß-carboxylic acid methyl ester of mp 235-237 ° C (identical to that prepared according to example 3 Connection).

Example 9

A culture medium consisting of 0.15% corn steep, 0.5% peptone and 0.5% glucose in distilled water, pH 7.3 is inoculated with Arthrobacter simplex ATCC 6946. The culture is allowed to grow at 28 ° for 24 hours and then a solution of 25 mg of 1 liter, 17aa-dihydroxy-3-oxo-D-homoandrost-4-en-17ass-carboxylic acid in 1 ml of aqueous methanol is added. After an incubation period of 48-72 hours, the mycelium is separated from the substrate and washed with water. The washing water combined with the substrate is extracted with methylene chloride. Workup of the extract yielded 1 lß, 17aa-dihydroxy-3-oxo-D-homoandrosta-l, 4-diene-17ass-carboxylic acid (identical to the product obtained in Example 1).

Example 10

1.05 g of 11β-hydroxy-3-oxo-17aa-propionyloxy-D-homodandrost-4-en-17ass-carboxylic acid methyl ester in 10 ml of methyl orthoformate and 10 ml of methanol was mixed with 10 ml of p-toluenesulfonic acid in a little methanol and 10 Minutes at room temperature. After adding 2 drops of pyridine, the mixture was poured onto dilute sodium hydrogen carbonate solution and extracted with methylene chloride. The methylene chloride extracts were washed with dilute saline, dried and evaporated. The crude reaction product was gassed with perchloryl fluoride in 35 ml DMF and 3.5 ml water at 0 ° C for 20 minutes. The mixture was poured onto water and extracted with methylene chloride. The extracts were washed with dilute sodium chloride solution, dried and evaporated, the residue was dissolved in 40 ml of glacial acetic acid, mixed with 0.4 ml of 30% hydrogen bromide in glacial acetic acid and left to stand at room temperature for 1 hour. Then it was poured onto ice water and extracted with methylene chloride, washed neutral with dilute sodium hydrogen carbonate solution and dilute sodium chloride solution, dried and evaporated. Chromatography on silica gel yielded 6a-fluoro-1 lß-hydroxy-3-oxo-17aa-propionyloxy-D-homopregn-4-en-1 7 aß-carboxylic acid methyl ester, UV: 8234 = 15900.

Example 11

200 ml of 17aa-acetoxy-1-beta-hydroxy-3-oxo-D-homoandrosta-1,4-diene-1 7-carboxylic acid methyl ester and 200 mg of tris (triphenylphosphine) rhodium (I) chloride were added under hydrogen in 4 ml of benzene and 4 ml of alcohol dissolved and hydrogenated overnight at 20 ° C. Chromatography on silica gel yielded 17 aa-acetoxy-11β-hydroxy-3-oxo-D-homoandrost-4-en-l 7 aß-car-bonsäuremethylester, mp. 235-237 ° C (ether); UV: £ 242 = 15 750; [a] o = + 49 ° (c = 0.1% in dioxane).

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