DE2203535A1 - 16-alpha-methyl-6, 20-dioxopregnane derivs - useful as inters for active steroids - Google Patents

Abstract

Cpds. of formula (I) : (where X is OH and Y is H or X + Y = O; R1 and R2 are H or OH), are prepd. by fermentation of a cpd. (II): (where R3 is H or lower acyl: R4 is H, OH or lower acyloxy) with a fungal culture of the genus Curcularia, pref. C. lunata. Cpds. (I) are new cpds. useful as inters. for pharmacologically active steroids.

Description

Process for the production of 6-keto steroids The invention relates to a process for the production of 6-keto steroids of the general formula I. in which X is a hydroxyl group and Y is a hydrogen atom or X and Y together are an oxo group and R1 and R2 are a hydrogen atom or a hydroxyl group, characterized in that an 85-steroid of the general formula II in which R1 has the same meaning as in formula I and R3 represents a hydrogen atom or a lower acyl group and R4 represents a hydrogen atom, a hydroxyl group or a lower acyloxy group, fermented with a mushroom culture of the genus Curvularia.

The invention also relates to the products of the general process Formula I.

The process according to the invention is carried out under the same reaction conditions carried out, which is usually carried out in the fermentation with fungal cultures of the genus Curvularia applies.

The most favorable ones are thus obtained in the generally customary preliminary tests Reaction conditions determined for aerobic, submerged fermentation. The transformation of the as a solution or Suspension of added substrate is determined by thin-layer chromatographic analysis followed by sample extracts. After the fermentation has ended, the process product becomes with a suitable water-immiscible solvent from the culture broth extracted and from the extract, e.g. B. by evaporation and other known work-up methods, for example by recrystallization and / or chromatography on silica gel.

If the starting compounds of the general formula II contain acyl groups, such as B. the formyl, acetyl, propionyl or butyryl group in the 3- and / or 21 position, these are saponified during fermentation.

Both the 3-hydroxy and the 3-ketosteroids can be used during fermentation of the general formula I are formed. Whether preferred the 3-ketosteroids or the 3-Hydroxysteroids are formed; this is due to the specific structure of the one used Output connection dependent.

The fermentation is preferably carried out using the curvularia mushroom species lunata carried out.

It is known that fermentation is used in steroid chemistry Fungal cultures of the genus Curvularia, especially with Curvularia lunata, used to in order to introduce an 11ß- and / or 14a-hydroxy group in steroids. It is therefore Surprisingly, in the fermentation of the starting compounds of the general Formula II 6-ketosteroids of the general formula I obtained, which neither in the llß- still carry a hydroxyl group in the 14a position.

The 6-ketosteroids of the general formula I are valuable intermediate products for the synthesis of pharmacologically active steroids, such as. B. 6-methyl steroids.

So you can, for example, the 6-ketosteroids, if necessary after selective reduction of a 3-keto group with lithium tri-tert. - bitoxyaluminum hydride and selective ketalization of the 20-keto group, by Grignardation into the corresponding Convert 6-methyl steroids. After the 3-hydroxy group has been oxidized and the 6-hydroxy group gives the corresponding 6 ", 16a-dimethyl-4-pregnen-3,20-dione. These can be converted in a known manner into the gestagen-active 17a-acyloxy-6a, 16a-dimethyl-4-pregnen-3,20-diones, such as B. 17a-acetoxy-6a, 16a-dimethyl-4-pregnen-3,20-dione, or in the anti-inflammatory Corticoids such as B. 11ß, 21-dihydroxy-6a, 16a-dimethyl-114-pregnadiene-3,20-dione be convicted.

The following examples serve to illustrate the invention Procedure.

B e i 5 p i e 1 1: A 2 l Erlenmeyer flask, of 500 ml for 30 minutes Nutrient solution of 1% corn steep liquor, 1% sterilized in an autoclave at 120 ° C Soya powder and 0.005% soybean oil, adjusted to pH 6.2, is made with a lyophil culture inoculated by Curvularia lunata and 72 hours at 30 ° C on a rotary shaker with a frequency of 145 rpm. shaken.

This preculture is used to inoculate a 20 l stainless steel fermenter Stahl, the 15 l of a medium sterilized at 121 oO from 1% corn steep liquor, Contains 0.5% starch sugar and 0.005% soybean oil (adjusted to pH 6.2).

With the addition of Silicon SH as an antifoam agent, at 29 0C below Aeration (10 l / min.) 0.7 atmospheric pressure and stirring (220 rpm) germinated for 24 hours.

1 liter of the culture broth is poured into 14 l of a Medium sterilized as above from 1% corn steep liquor, 1.25% soy powder and 0.005% Soybean oil transferred and grown under the same conditions.

After 6 hours, a solution of 7.5 g of 3-hydroxy-16-methyl-5-pregnen-20-one is obtained added in 200 ml of glycol monomethyl ether and 48 1/2 under the same conditions hours fermented.

The culture broth is then after separating the mycelium with methyl isobutyl ketone extracted.

The extract is concentrated at 30 ° C. in vacuo. The concentrate is concentrated to dryness in a rotary evaporator at a bath temperature of 30 ° C and the The residue was washed out with 100 ml of hot hexane. The crude product 6.7 g is on a Chromatographed silica gel column using acetone-hexane (1: 1).

2.8 g of 3a-hydroxy-16a-methyl-5ß-pregnane-6,20-dione are obtained, which can be recrystallized from ethyl acetate.

Melting point: 186/188 - 1900 Example 1 2: Under the conditions of Example 1, 18 g of 3β-hydroxy-21-acetoxy-16a-methyl-5-pregnen-20-one are dissolved Fermented in 100 ml glycol monomethyl ether with Curvularia lunata for 30 hours and as described worked up. The crude product is purified by column chromatography Purified on silica gel using an acetone-hexane gradient and separated into two components. The first eluates contain 2 g of 21-hydroxy-16a-methyl-5ß-pregnane-3,6,20-trione, which is recrystallized from ethyl acetate.

Melting point: 199-200 ° The following eluates provide 2 g of 3a, 21-dihydroxy-16amethyl-5ß-pregnane-6,20-dione, which is also recrystallized from ethyl acetate.

Melting point: 190-193 °

Claims (6)

P atentane spray b;) Process for the preparation of 6-ketosteroids of the general formula I in which 1 is a hydroxyl group and Y is a hydrogen atom or X and Y together are an oxo group and R1 and R2 are a hydrogen atom or a hydroxyl group, characterized in that an A5 steroid of the general formula II where R1 has the same meaning as in formula I and R3 represents a hydrogen atom or a lower acyl group and R4 represents a hydrogen atom, a hydroxyl group or a lower acyloxy group, fermented with a mushroom culture of Gatturg Curvularia. 2.) The method according to claim 1, characterized in that one Fungal culture of the species Curvularia lunata is used. 3.) 6-ketosteroids of the general formula I. where X is a hydroxyl group and Y is a hydrogen atom or X and Y together are an oxo group and R1 and R2 are a hydrogen atom or a hydroxyl group. 4.) 3α-Hydroxy-16α-methyl-5β-pregnane-6,20-dione. 5.) 3α, 21-dihydroxy-16α-methyl-5β-pregnane-6,20-dione. 6.) 21-Hydroxy-16a-methyl-5ß-pregnane-3,6,20-trione.