JPH0370720B2 - - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to the general formula (): [In the formula, X represents an oxo group or a propylene dioxy group, △ represents a carbon atom bonded to an adjacent carbon atom through one double bond and two single bonds, and R ₁ represents a methyl group. Regarding the production of an estrene-3,17-dione derivative of
This is the general formula (): [In the formula, R ₁ represents the same thing as above] is fermented using a Penicillium genus bacterial culture, resulting in a general formula (): ketalizing the 15α-hydroxysteroid of the formula in which R ₁ is as defined above with an alkanediol or o-diphenol having 2 to 6 carbon atoms in any order in the presence of an acidic catalyst; and acylated with sulfonic acid chloride in the presence of a base, and the obtained general formula (): [In the formula, △ and R ₁ represent the same as above,
X' represents an alkylenedioxy group or o-phenylenedioxy group having 2 to 6 carbon atoms, and Y represents an alkylsulfonyl group or an arylsulfonyl group] by treating with a base a compound of the general formula ( ) to the ^Δ15 -steroid and, if desired, by hydrolyzing the obtained ketal by known methods.

Estrene-3,17-dione derivatives of the general formula () are substances with known pharmacological effects and/or are important intermediates for producing pharmaceutically effective steroids, This could only be produced by the conventionally known method using a very expensive multi-step synthesis method [West German Patent Application No. 1593500, ibid.
1593501 and 1643050 and J. Fried and J.A. Edwards, Organic Reaction in Steroid Chemistry
Steroid Chemistry; Van Nostrand
Reinhold Comp.New York) (1972)
See Volume 1, pages 301 et seq.].

On the contrary, the process of the invention has the advantage that estrene-3,17-dione derivatives of the general formula can be prepared in a simpler manner and with higher yields than is possible using known processes.

In the first step of the process of the invention, the steroid of the general formula is fermented with a Penicillium fungus culture. For this fermentation, the following strains are suitable, for example: Penicillium canescens
canescens) (ATCC 10419), Penicillium
Chrysogenum (Penicillium chrysogenum)
(ATCC 10003), Penicillium nigricans (ATCC 9439), Penicillium stronifelum (Penicillium
stoloniferum (ATCC 14586) or especially Penicillium raistritskii
raistrickii) (ATCC 10490). Fermentations using this strain are carried out under conditions customarily used during microbiological hydroxylation of steroids using fungal cultures.

Under the above conditions, steroids of the general formula can be hydroxylated to 15α-hydroxysteroids of the general formula in surprisingly high yields.

This favorable result was unexpected. This is because known Penicillium bacteria not only have the ability to hydroxylate steroids at the 15α-position, but also have the ability to hydroxylate steroids at the 6β-position or the 11α-position.
This is because it is also possible to hydroxylate at the - position and reduce the ketosteroid to the corresponding hydroxysteroid.

In the second and third steps of the process of the invention, the 15α-hydroxysteroids of general formula () are ketalized with alkanediols or o-diphenols in any order and acylated with sulfonic acid chlorides. Ru.

Selective ketalization typically involves 3-keto-△ ⁴ −
It is carried out under conditions used for ketalization of steroids. For example, steroids and alkanediols (e.g. glycols, 1,3-propanediol, 2,3-butanediol or 2,2-
dimethylpropanediol) or o-diphenyrol in an insoluble solvent (e.g. benzole, chloroform, methylene chloride, tetrachloroethane, diethyl ether or tetrahydrofuran) with a catalyst (e.g. hydrogen chloride, sulfuric acid, perchloric acid, trifluoroacetic acid, p- Toluolsulfonic acid) can be used for the reaction. In order to obtain high yields of ketalized products, water binders such as calcium sulfate, magnesium sulfate, molecular sieves or formic acid trialkyl esters are preferably added to the reaction mixture.

During this ketalization, it is in principle possible to also ketalize the 17-keto group under aggressive reaction conditions and over long reaction times. In order to avoid this, it is advantageous to determine the optimum reaction time in the usual manner in preliminary experiments.

During this ketalization reaction, the △ ⁴ -double bond present in the steroid is transferred to the △ ⁵⁽⁶⁾ -position or △
⁵⁽¹⁰⁾ Moved to − position. This double bond transition is
It is not critical to the implementation of the method of the invention. This is because during the subsequent ketal decomposition, which is always necessary for the production of pharmaceutically effective steroids, steroids with the 3-keto-△ ⁴ -structure are again obtained. It is.

15α- using alkylsulfonic acid chloride
Acylation of the hydroxy group is carried out using known procedures for this purpose. For example, a 15α-hydroxysteroid can be combined with a sulfonic acid chloride (e.g. benzenesulfonic acid chloride, p-toluolsulfonic acid chloride or especially methanesulfonic acid chloride) and a tertiary amine (e.g. pyridine, lutidine, collidine or 4-dimethylaminopyridine). ) can cause a reaction.

The compound of general formula () obtained in this way is
It can be converted into the ^Δ15 -steroid of the general formula () by treatment with a base.

Removal of the sulfonic acid group is carried out in the presence of a base. Suitable bases are, for example, alkali metal hydroxides, alkaline earth metal hydroxides, alkali metal carbonates, alkali metal salts of lower carboxylic acids or tertiary amines.

Examples of suitable bases include: sodium hydroxide, potassium hydroxide, calcium hydroxide, sodium carbonate, potassium carbonate, sodium acetate, potassium acetate, pyridine, lutidine or collidine. When using tertiary amines for this reaction, they simultaneously serve as solvents, and when using inorganic bases,
The reaction is preferably carried out in dipolar and neutral solvents such as dimethylformamide, N-methylacetamide, N
- carried out in the presence of methylpyrrolidone, acetonitrile, dimethylsulfoxide or hexamethylphosphoric triamide.

For these removals, similar removals are known (see J.Amer.Chem.Soc.78 1956, p. 6347).
As in, there is a risk that the primarily formed △ ¹⁵ - double bond is isomerized and becomes a △ ¹⁴ - double bond. To avoid this, care should be taken to maintain exactly the optimum temperature and reaction time (these can be determined in preliminary experiments). When this removal is carried out using alkali metal salts of lower carboxylic acids (in particular sodium or potassium acetate) in dipolar neutral solvents at reaction temperatures between -20 °C and +40 °C, the isomerization There is no danger of Under these conditions, the sulfonic acid group is surprisingly not exchanged for the carboxylic acid residue of the base, but is removed under the formation of a Δ ¹⁵ -double bond.

If desired, the ketals obtained can be hydrolyzed in known manner by treatment with aqueous acids. On the other hand, the ketal itself is 17β-hydroxy-18-methyl-17α in later usage examples.
As detailed in the example of the synthesis of -ethynyl-15β,16β-methylene-4-estren-3-one, it can also be used as an intermediate for the production of pharmaceutically effective steroids.

The invention will now be explained with reference to examples.

Example 1 (a) Glucose 3.0%, corn staple 1.0, sterilized in an autoclave at 120°C for 30 minutes.
%, _NaNO3 0.2%, _{KH2PO4 0.1} %,
_{K2HPO4 0.2} %, _MgSO4 0.05%, _FeSO4 0.002%
A 2 l Erlenmeyer flask with 500 ml of nutrient solution containing 0.05% KCl is inoculated with a freeze-dried culture of Penicillium leistritskii (ATCC 10490) and shaken on a rotary stirrer for 72 hours at 30°C. Then 250 ml of this preculture was added to
Media of the same composition sterilized at 120°C and 1.1 atu¨15
Inoculate a 20 l glass fermenter filled with With the addition of silicone SH as a foam suppressant, at 29°C and with air (10/min) blowing, 0.7 atu¨pressure and stirring (220 revolutions/min).
Ferment for 24 hours.

The culture solution 1.8 is transferred under aseptic conditions into a similarly sterilized nutrient medium 26 of the same composition as the culture medium and fermented under the same conditions as the prefermentation culture. After 12 hours, nat.-18-methyl-4-
Estrene-3,17-dione (where nat.
120 g of sterile subdivided suspension (meaning this steroid has natural steroid coordination) 2
Add and further ferment.

The progress of the conversion is followed by thin layer chromatographic analysis of the methyl isobutyl ketone extract of the fermentation sample. After about 70 hours of contact time, the conversion is complete. The micelles are then filtered off and the culture solution is extracted twice with 200 ml of methyl isobutyl ketone. In parallel, the micelles filtered off are vigorously stirred with a mixture of methyl isobutyl ketone, acetone and water and extracted until the steroid substances can no longer be detected.

The organic extract solutions are combined and evaporated to dryness in vacuo at a bath temperature of 50°C. The brown crystalline residue is washed several times with hexane to remove the silicone oil, dried and finally, after treatment with activated carbon, recrystallized from acetic acid ester to give pure nat.- with a melting point of 175-177 °C. 15α-hydroxy-18-methyl-4-
97.3 g of estrene-3,17-dione (theoretical amount)
76.5%).

(b) Methylene chloride in 32 g of nat.15α-hydroxy-18-methyl-4-estrene-3,17-dione
In 240 ml and 64 ml of o-formic acid ethyl ester,
2,2-dimethyl-1,3-propanediol
Add 96 g and 320 mg of p-toluolsulfonic acid and stir for 30 minutes at 50° C. under nitrogen atmosphere. It is then diluted with ether, washed with sodium bicarbonate solution and water, dried and evaporated to dryness. The residue was chromatographed on silica gel and the nat.15α-hydroxy-3,3-(2′,
2'-dimethyl-1',3'-propylenedioxy)-
37 g of 18-methyl-5- or -5(10)-estren-17-one are obtained as an oil.

(c) nat.15α-hydroxy-3,3-(2',2'-dimethyl-1',3'-propylenedioxy)-18-
Methyl-5- or -5(10)-estrene-17
27.1 ml of methylene sulfochloride was added to 37 g of -one in 370 ml of pyridine under ice cooling, and the mixture was stirred at ice bath temperature for 3 hours. Then stirred and introduced into ice water,
The precipitate is aspirated, washed thoroughly with water, then taken up in methylene chloride and dried. nat.15α−
Mesyloxy-3,3-(2',2'-dimethyl-1',
3′-propylenedioxy)-18-methyl-5-
Alternatively, -5(10)-estren-17-one is obtained as an oil.

(d) nat.−15α-mesyloxy-3,3-(2′,2′-
dimethyl-1',3'-propylenedioxy)-18
-Methyl-5- or -5(10)-estrene-
35 g of 17-one was dissolved in 350 ml of dimethylformamide with 105 g of anhydrous sodium acetate at room temperature for 20 min.
Stir for an hour. Next, stir and introduce into ice water,
The resulting precipitate is aspirated, washed and taken up in methylene chloride. After evaporative concentration crude nat.3,3
-(2',2'-dimethyl-1',3'-propylenedioxy)-18-methyl-5- or -5(10),15
28.9 g of -estradien-17-one are obtained.

Claims (1)

[Claims] 1 General formula (): [In the formula, X represents an oxo group or a propylene dioxy group, △ represents a carbon atom bonded to an adjacent carbon atom through one double bond and two single bonds, and R ₁ represents a methyl group. To prepare the estrene-3,17-dione derivative of the general formula (): [In the formula, R ₁ represents the same thing as above] is fermented using a Penicillium genus bacterial culture, resulting in a general formula (): ketalizing the 15α-hydroxysteroid of the formula in which R ₁ is as defined above with an alkanediol or o-diphenol having 2 to 6 carbon atoms in any order in the presence of an acidic catalyst; and acylated with sulfonic acid chloride in the presence of a base, and the obtained general formula (): [In the formula, △ and R ₁ represent the same as above,
X′ represents a propylene dioxy group and Y represents a mesyloxy group] is converted into a △ ¹⁵ -steroid of the general formula () by treating it with a base, and if desired, the obtained ketal can be converted into a △ 15 -steroid of the general formula () A method for producing an estrene-3,17-dione derivative, which comprises hydrolyzing it by a method.