AT253135B - Process for making new steroid compounds - Google Patents

Description

  

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  Process for making new steroid compounds
The invention relates to a process for the preparation of new N-substituted 4, 6-pregnadieno- [3, 2-c] - pyrazole compounds and in particular new 21-hydroxy-, 21-acyloxy- and 17ot-hydroxy-20-oxo- 4, 6-pregnadieno- [3, 2-c] -pyrazoles in which a hydrocarbon substituent or a substituted hydrocarbon substituent is bonded to a nitrogen atom of the pyrazole nucleus.



   The new 4,6-pregnadieno- [3, 2-c] pyrazole compounds obtainable according to the invention have the general formula
 EMI1.1
 in which Rl is an 8-hydroxy or keto radical or an β-halogen radical, but the latter only if X is halogen, R2 is fluorine, hydrogen or a methyl group, Rg is a hydrogen atom, an α-methyl, β-methyl or Methylene group, R4 a hydroxyl group, an acyloxy group containing 1-10 carbon atoms or a phosphoryloxy group and Rg an alkyl, cycloalkyl, aralkyl or aryl group, while X denotes a hydrogen or halogen atom.

   N-substituted pyrazole compounds of the formula A are hereinafter referred to as 1'-substituted 4, 6-pregnadieno- [3, -pyrazole, while N-substituted pyrazole compounds of the formula B are referred to as 2'-substituted 4, 6-pregnadieno- [ 3, 2-c] -pyrazoles are designated.



   The 4,6-pregnadieno- [3, 2-c] -pyrazoles defined above, obtainable according to the invention, have a strong anti-inflammatory activity and are particularly effective for the treatment of arthritis and the like. similar Diseases in that they can be administered in low doses to exert their cortisone-like effect, which reduces the occurrence of undesirable side effects.



   In the process according to the invention for the preparation of these new chemical compounds, compounds of the general formula are used as starting materials

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 EMI2.1
 are used in which Rl, R, Ra and X have the above meaning, Rg is hydrogen or an alkyl group, and in which the oxygen functions in C-17, C-20 and C-21 with suitable blocking groups, such as 17, 20, 20 , 21-bis (methylenedioxy), can be linked.



   The compounds used as starting materials in the process according to the invention can be obtained, for example, starting from a 17 a, 21-dihydroxy-4, 6-pregnadiene-3, 20-dione which has the following general formula
 EMI2.2
 can be represented in which Rti is an ß-hydroxy group or an ß-halogen substituent, but the latter only when X is halogen and R, R and X have the meaning given above.



   It has been found that the above-defined compounds of the general formula IV with aqueous formaldehyde solutions in the presence of a strong acid to form compounds of the general formula
 EMI2.3
 react, in which Ril, R, R. and X have the above meanings. For example, cold concentrated hydrochloric acid and formalin are added with stirring to a suspension of the steroid in chloroform that is cooled to about OOC. The mixture is then allowed to reach room temperature and is stirred for several hours. where the corresponding 17a. 20. 20. 21-bis (methylenedioxy) derivative is formed.

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   The 17 <x, 20, 20, 21-bis (methylenedioxy) -4,6-pregnadiene-3,11-dione compound of the general formula
 EMI3.1
 in which R-, Rg and X have the above meanings, oxidation of the corresponding liss-hydroxy-17 a, 20, 20, 21-bis (methylenedioxy) -4, 6-pregnadien-3-one, z . B. with chromium trioxide in an anhydrous base such as pyridine. If desired, however, the 17α, 20,20,21-bis (methylenedioxy) -4, 6-pregnadiene-3, 11-dione can also be converted directly by reaction of the 17a. 21-dihydroxy- 4, 6-pregnadiene-3, 11, 20-trione can be prepared with formaldehyde solution in the presence of an acid, as described above.



   In the reaction of an 11ss-hydroxy-, 11ss-halogen- or 11-keto-17α, 20,20,21-bis (methylene dioxy) -4, 6-pregnadien-3-one compound with an alkyl formate and Sodium hydride in an inert atmosphere forms the corresponding 17α, 20,20,21-bis- (methylenedioxy) -2-hydroxymethylene-4, 6-pregnadien-3-one of the general formula
 EMI3.2
 in which R1, R2, R3 and X have the above meanings. The steroid is preferably dissolved in a solvent such as benzene, and the solution is cooled to room temperature and ethyl formate is added. The air in the system is displaced with nitrogen, sodium hydride is added and the mixture is stirred for several hours at room temperature.



   By reacting a 17 a, 20,20, 21-bis (methylenedioxy) -2-hydroxymethylene-4, 6-pregnadien- - 3-one compound with a lower alkanol in the presence of an acidic reagent such as p-toluenesulfonic acid, can the corresponding 17α, 20,20,21-bis (methylenedioxy) -2-alkoxymethylene-4,6-pregnadien- -3-one can be obtained.



   The inventive method for the preparation of the new steroid compounds of the Pregnanreihen of the general formula I consists in its essence that one compound of the above general formula n, in which Rl - Rs, R6 and X have the meaning given, with an alkyl, cycloalkyl -, aralkyl-, arylhydrazine or a salt of such a compound, the corresponding N-substituted 4, 6-pregnadieno [3, 2-c] pyrazole compounds of the general formula

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 EMI4.1
 
 EMI4.2
 becomes
 EMI4.3
 
The resulting products can be separated from one another by chromatography.



   When a 17a, 20, 20, 21-bis (methylenedioxy) -2-hydroxymethylene - 4, 6 - pregnadien - 3 - one compound is reacted directly with an aryl hydrazine (without formation of the 2-alkoxymethylene derivative), a Isomer in predominant amount, while in the implementation of the 17 a, 20, 20, 21-bis (methylenedioxy) -2-alkoxymethylene-4, 6-pregnadien-3-one compound with an aryl hydrazine considerable amounts of both isomers are formed.



   When this reaction is carried out with alkylhydrazines, mixtures can be obtained both when starting from the 2-hydroxymethylene steroid and when starting from the 2-alkoxymethylene steroid. A mixture of isomers can also arise in the reaction of an alkyl, cycloalkyl, aralkyl, aryl hydrazine with a 2-hydroxymethylene compound, which as a result of the recovery process used, e.g. B. as a result of recrystallization in the presence of a trace of alcohol from a solution of the 2-hydroxymethylene compound which has not been completely freed from acid, contains changing amounts of the 2-alkoxymethylene derivative.



   Examples of the alkyl hydrazines which can be used in the process according to the invention are methyl hydrazine, ethyl hydrazine, propyl hydrazine, butyl hydrazine, β-hydroxyethyl hydrazine; Examples of arylhydrazines, phenylhydrazines and the substituted phenylhydrazines are o-, m- and p-halophenylhydrazines, o-, m- and p-toluylhydrazines, 0-, m- and p-alkoxyphenylhydrazines, 0-, m- and p - Ni trophenylhydrazine, 1-hydrazinonaphthalene, 2-hydrazinopyridine, 3-hydrazinopyridine, 4-hydrazinopyridine, 4-hydrazinopyridine oxide and 2-hydrazinopyrimidine;

     Examples of aralkyl hydrazines are benzyl hydrazine and phenylethylene hydrazine.
 EMI4.4
 

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 EMI5.1
 
In treating pyrazole compounds obtained after the reaction with an alkylhydrazine, cycloalkylhydrazine, aralkylhydrazine, arylhydrazine or a salt of such a compound having blocking groups at C-17, C-20 and C-21, e.g. Of 17α, 20,20,21-bis (methylenedioxy) -4,6-pregna-dieno [3, 2-c] pyrazole compounds with hydrolyzing agents such as a dilute organic acid, e.g.

   B. with 60% aqueous formic acid, the blocking groups in C-17,
 EMI5.2
 Protective 17 a, 20, 20, 21-bis (methylenedioxy) group, split off and reduced manganese-llss-hydroxy-4,6-pregnadieno [3, 2-c] pyrazole compound, e.g. B. by adding a saturated solution of sodium borohydride to a solution of the steroid in a mixture of triethylamine and isopropyl alcohol, to which a little water is preferably added, and the mixture is left to stand overnight.



   Acyl groups on the R4 residue and / or on the R- residue can be removed by reacting the steroid with sodium methylate in methanol at room temperature, the corresponding 17 a, 21-dihydroxy-20-oxo-4,6-pregnadieno- [3 , 2-c] -pyrazole forms. Acyl groups on the R radical can be split off selectively by treatment with aqueous acetic acid.



   The 21-acylates of 17α, 21-dihydroxy-20-oxo-4,6-pregnadieno- [3, -pyrazoles obtained as described above can be prepared by reacting the N-substituted 17a, 21-dihydroxy-20-oxo-4 , 6-pregnadieno- [3, 2-cl-pyrazole can be prepared with an acylating agent. Acylating agents which can be used for this purpose are e.g. B. from lower hydrocarbon carboxylic acids with 1 to 10 carbon atoms derived acylating agents such as benzoic anhydride or tert. Butylacetyl chloride, lower aliphatic carboxylic acid anhydrides, such as acetic acid anhydride or propionic acid anhydride, or polycarboxylic acid anhydrides, such as ss. ss-dimethylglutaric anhydride, succinic anhydride and the like like

   The acylation takes place in the presence of an organic base such as pyridine.



   The 21-phosphoric acid monoesters can be prepared by reacting the corresponding 21-iodine compound with a mixture of silver phosphate and phosphoric acid. Both the mono- and dialkali salts of these esters can be obtained by neutralizing the phosphoric acid monoester with an alkali metal methylate.



   Those 4, 6-Pregnadieno- [3, 2-c] -pyrazoles, which at the 1'- or the 2'-position nitrogen
 EMI5.3
 



   The following examples are intended to explain the invention further, but not to limit its scope.



     Example 1: 200 mg of 17α, 20,20,21-bis (methylenedioxy) -11ss-hydroxy-2-hydroxymethylene-16α - - methyl-4,6-pregnadien-3-one in 7 ml of absolute ethanol are mixed with 82 mg of sodium acetate and then 102 g of cyclohexyl hydrazine oxalate were added. The mixture is refluxed for 1 hour under nitrogen. The insolubles are filtered off and the filtrate is evaporated to dryness. The residue is dissolved in 3 ml of ether and the ethereal solution is first washed with 2% aqueous sodium hydroxide solution and then washed neutral with water. The ether solution is then dried over magnesium sulphate, filtered and evaporated to dryness.

   The main component of the residue is N-cyclohexyl-17a, 20.20, 21-bis- (methylenedioxy) -11ss-hydroxy-16α-methyl-4,6-pregnadieno [3,2-c] pyrazole.



   According to another method, a mixture of l'-cyclohexyl and 2'-cyclohexyl
 EMI5.4
 

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 - (methylenedioxy) -116-hydroxy-2-methoxymethylene-16ct-methyl-4, 6-pregnadien-3-one, which is then reacted with cyclohexylhydrazine according to the process described in detail in Example 2 by adding cyclohexylhydrazine in the second process stage Instead of phenylhydrazine is used.



  The components of the mixture are separated from one another by chromatography.
 EMI6.1
 heats. The mixture is evaporated to dryness and then treated with water. The product is extracted with chloroform and the chloroform solution is washed with saturated aqueous sodium chloride solution, 5% aqueous sodium bicarbonate solution and then with water. The chloroform solution is dried over magnesium sulfate and then evaporated to dryness. The residue is taken up in ether which contains some methanol, stirred with 75 mg of decolorizing charcoal, filtered and the filtrate is added
 EMI6.2
 receives N-cyclohexyl-llss, 17 <x, 21-trihydroxy-16a-methyl-20-oxo-4,6-preg-converted.



     If the above method is based on 2l-cyclohexyl-17a, 20, 20, 21-bis (methylenedioxy) -llss-hydroxy-16a-methyl-4,6-pregnadieno- [3, 2-c] pyrazole if the 1'-cyclohexyl derivative is used instead of the 2'-cyclohexyl derivative, the corresponding 1'-cyclohexyl compounds are obtained.



   If one proceeds in all of the above processes from the 17a, 20, 20, 21-bis (methylenedioxy) -2-hydroxymethylene 6-pregnadiene derivative, which is obtained from one of the starting materials listed at the beginning of the description, one obtains the corresponding l'-cyclohexyl and 2'-cyclohexyl derivatives.



   If other cycloalkylhydrazines are used instead of the cyclohexylhydrazine, the corresponding 11- and 21-cycloalkyl derivatives are obtained by the above processes.



   Example 2: To a suspension, cooled to 5 ° C in an ice bath, of 25.0 g of 11ss, 17α21-trihydroxy-6,16α-dimethyl-4,6-pregnadiene-3,20-dione in 1.5 l of non-alcoholic chloroform 750 ml of cold concentrated hydrochloric acid and then 750 ml of formalin (with a low methanol content) are added with constant stirring. The mixture is removed from the ice bath and stirred for 7 hours at room temperature. The layers are separated and the aqueous phase is extracted twice with chloroform. The combined organic layers are washed twice with 5% sodium carbonate solution and twice with saturated saline solution. The solution is dried over magnesium sulphate and evaporated to dryness under reduced pressure.

   The residue is stirred with methanol, whereupon a crystalline solid is obtained. According to paper strip chromatography, this material no longer contains any detectable amounts of starting material, but shows two ultraviolet absorption sites near the solvent front (methanol-formamide 2: 1 versus benzene-n-hexane 1: 1). 2, 425 g
 EMI6.3
 stage is used without further purification.



   500 mg of 17 a, 20.20, 21-bis (methylenedioxy) -11ss-hydroxy-6,16? -Dimethyl-4,6-pregnadien-3-one are dissolved in 25 ml of benzene, followed by 5 ml of benzene at normal Pressure to be distilled off. The solution thus obtained is cooled to room temperature. Then 0.75 ml of freshly distilled ethyl formate are added, the air is displaced by nitrogen, and 150 mg of sodium hydride (as a 57% dispersion in mineral oil) are added. The mixture is stirred for 3 hours under nitrogen at room temperature. 15 ml of saturated aqueous monosodium phosphate solution are then added and the product is extracted with ether. The ether extracts are extracted with 2N sodium hydroxide solution and the sodium hydroxide extracts are acidified with monosodium phosphate and extracted again with ether.

   After evaporation of the ether extract to dryness, 500 mg of crude product are obtained. 290 mg of 17a, 20, 20, 21-bis (methylenedioxy) -11ss-formyloxy-2-hydroxymethylene-6,16α-dimethyl-4,6-pregna-dien-3-one are obtained from the ether solution in the form of yellow crystals won; Mp = 220-2360C. The analytical sample is made from ethyl
 EMI6.4
 
 EMI6.5
 

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 EMI7.1
   -2170Cj ultraredmaximabisb, x andb, 4.



   1. 19 g of 17ct, 20, 20, 21-bis (methylenedioxy) -llss-hydroxy-2-hydroxymethylene-6, 16a-dimethyl- - 4. 6-pregnadien-3-one are dissolved in 25 ml of ethanol. 300 mg of phenylhydrazine are added and the mixture is heated to reflux temperature under nitrogen for 1 hour. After adding 25 ml of water, the product is extracted with 150 ml of ether. The extracts are washed with 2N hydrochloric acid, saturated sodium bicarbonate solution, water and saturated sodium chloride solution, dried over sodium sulfate and evaporated to dryness. 1.2 g of crude product are obtained. After recrystallization from ether, 17ct-20, 20, 21-bis (methylenedioxy) -llss-hydroxy-6, 16a-dimethyl-2'-phenyl-4, 6-pregna- dieno- [3, 2- c] pyrazole.



   In another process, a mixture of 1'-phenyl- and 2'-phenyl-17 <x, 20, 20, 21-bis (methylenedioxy) -llss-hydroxy-6, 16ct-dimethyl-4,6-pregnadieno - [3, 2-c] -pyrazole prepared as follows: A mixture of 1 17?, 20,21,21-bis (methylenedioxy) -11ss-hydroxy-2-hydroxymethylene-6,16? -Dimethyl-4, 6-pregnadien-3-one, 200 ml of methanol and 200 mg of p-toluenesulfonic acid are heated to reflux temperature and then left to stand for 1 hour at room temperature. The reaction mixture is diluted with water and extracted with ethyl acetate. The ethyl acetate extract is washed twice with 2N aqueous sodium hydroxide solution and then with water, dried and concentrated in vacuo.



   The 17α, 20,21,21-bis (methylenedioxy) -11ss-hydroxy-2-methoxymethylene-6,16α-dimethyl-4,6-pregnadien-3-one is acidified by direct crystallization or by chromatography washed aluminum oxide and eluting with mixtures of ether and chloroform.



   A mixture of 500 mg of 17α, 20,20,21-bis (methylenedioxy) -11ss-hydroxy-2-methoxymethylene-6, 16a-dimethyl-4. 6-pregnadien-3-one, 100 ml of ethanol and 1 ml of phenylhydrazine is heated under nitrogen until solution occurs and then left to stand overnight at room temperature under nitrogen.



  2 ml of acetic acid are added and the mixture is left to stand for a further 4 hours. The reaction mixture is then diluted with ethyl acetate, washed twice with 2N sulfuric acid, twice with 2.5N sodium hydroxide solution and twice with water. The ethyl acetate extract is dried, concentrated and chromatographed on aluminum oxide washed with acid.



   By eluting with benzene, a product is obtained which, when recrystallized from a mixture
 EMI7.2
 Washed water, then dried over sodium sulfate and evaporated in vacuo to give 430 mg of crude product. This is dissolved in 60 ml of absolute methanol and 0.1 equivalent of sodium methylate in methanol is added to the solution. The mixture is stirred for 15 min under nitrogen at room temperature and then acidified with acetic acid, whereupon the solvent is driven off in vacuo at room temperature. 20 ml of water are added and the product is extracted with 150 ml of ethyl acetate. The ethyl acetate solution is washed with saturated sodium bicarbonate solution and then with water, dried over sodium sulfate and evaporated to dryness. An amorphous solid is obtained.



   This crude product is dried in a high vacuum and dissolved in 4 ml of pyridine. 3 ml of acetic anhydride are added, the mixture is heated on the steam bath for 15 minutes and then evaporated to dryness in vacuo. Then 20 ml of water are added. The product is extracted with 150 ml of ethyl acetate, washed with saturated sodium bicarbonate solution and with water and dried over sodium sulfate.



  The residue obtained after evaporation of the solvent in vacuo becomes from a mixture
 EMI7.3
 razol-21-acetate are dissolved in 10 ml of absolute methanol, and the solution is treated with 1.1 equivalents of sodium methylate in methanol. The mixture is stirred for 15 minutes at room temperature under nitrogen. The product is acidified with acetic acid and then evaporated in vacuo at room temperature. The residue is taken up in 100 ml of chloroform, washed with water, saturated sodium bicarbonate solution and again with water, dried over sodium sulfate and dried to dryness

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 evaporated. 100 mg of 118, 17 ex, 21-trihydroxy-6, 16ex-dimethyl-20-oxo-21-phenyl-4, 6-pregnadieno [3, 2-c] pyrazole are obtained.



   The crude 118, 17ex, 21-trihydroxy-6, 16ex-dimethyl-20-oxo-2'-phenyl-4, 6-pregnadieno [3, 2-c] pyrazole is dissolved in 1.2 ml of dimethylformamide . The solution is cooled to OOC and 0.07 ml of methanesulfonyl chloride are added. The mixture is kept at OOC for 1 h, 3 ml of water are added and the product is extracted with 100 ml of chloroform, the extract is washed with water, dried over sodium sulfate and evaporated to dryness. Crude llss, 17a, 21-trihydroxy-6, 16ct-dimethyl- - 20-oxo-21-phenyl-4,6-pregnadieno- [3, 2-c] -pyrazole-21-mesylate are obtained.



   The llss, 17ct, 21-trihydroxy-6, 16K-dimethyl-20-oxo-21-phenyl-4, 6-pregnadieno- [3, 2-c] -pyrazole-21-mesylate is in 5.5 ml Suspended acetone and the suspension mixed with 117 mg of sodium iodide.



  The mixture is refluxed under nitrogen for 1 hour and then cooled on ice. When water is added, a white precipitate forms, which is filtered off, washed with water and dried in vacuo. 11ss, 17α-dihydroxy-21-iodo-6,16α-dimethyl-20-oxo-2'-phenyl-4,6-pregnadieno- [3, 2-c] pyrazole is obtained.



   This material is dissolved in 5 ml of ethanol. A solution of 500 mg of sodium bisulfite in 5 ml of water is added and the mixture is heated to reflux temperature for 1 hour under nitrogen. Then 10 ml of water are added and the product is extracted with 100 ml of chloroform, the extract is washed with water, dried over sodium sulfate and evaporated to dryness.



   The residue, which weighs 100 mg, is purified by chromatography on silica gel. The resulting llss, 17cx-dihydroxy-6, 16K-dimethyl-20-oxo-21-phenyl-4, 6-pregnadieno- [3, 2-e] -pyrazole (about 70 mg) is made from a mixture of acetone and Ether recrystallized.



   Monosilver phosphate is produced by reacting 32 g of trisilver phosphate with 10 μl of 100% phosphoric acid with thorough mixing in a three-necked round bottom flask. The monosilver phosphate is washed twice with diethyl ether and the diethyl ether is decanted to remove some of the phosphoric acid. The monosilver phosphate is covered with a layer of 200 ml of acetonitrile and the mixture is heated to reflux temperature. 20 gllss, 17c <-dihydroxy-21-iodo-6, 16 <x-dimethyl-20-oxo-21-phenyl-4, 6-pregnadieno [3, 2-c] -pyrazole are added, and the mixture is heated to reflux temperature for 75 minutes under nitrogen and with stirring.

   The reaction mixture is then cooled to room temperature over the course of 1 hour. 200 g of ice water are added and the acetonitrile is driven off in vacuo at a temperature below 250C. The pH of the aqueous suspension thus obtained is adjusted to 6.4 by adding 23 ml of saturated aqueous sodium bicarbonate solution. The precipitate that forms is filtered off and washed with water until no more ultraviolet-absorbing material can be found in the washing water. The filtrate and the washing water are combined together and subjected to freeze-drying to separate a solid material from the water. The solid material is stirred with a total of 770 ml of methanol in seven portions. The material insoluble in methanol is filtered off.

   The filtrate is concentrated to 200 ml in vacuo and poured through a column charged with 60 g of cation exchange resin ("IR-120") in the hydrogen form. The column is washed with methanol until the washing liquid no longer contains any ultraviolet-absorbing material. The eluate combined with the washing liquids is concentrated to 15 ml and mixed with 150 ml of ether. The precipitate is filtered off, washed with ether and left in a desiccator for 16 h.
 EMI8.1
 - [3, 2-c] pyrazole-21-dihydrogen phosphate.



   The mono- and dialkali salt of this 21-phosphoric acid monoester are obtained by neutralizing the ester with alkali metal methylate.



     A solution of 62mg 11ss, 17α, 21-trihydroxy-6,16α-dimethyl-20-oxo-2'-phenyl-4, 6-pregnadieno- [3, 2-c] -pyrazole-21-mesylate in 1 ml of freshly distilled anhydrous dimethylformamide is mixed with enough anhydrous potassium fluoride that a saturated solution is formed. The mixture is heated to 110 ° C. for 20 h. The cooled solution is mixed with water and the product is extracted with chloroform, the extract is dried over sodium sulfate and evaporated to dryness.

   The product is a mixture of 17α, 21-epoxy-11ss-hydroxy-6,16α-dimethyl-20-oxo-2'-phenyl-4, 6-pregnadieno- [3, 2-c] - - pyrazole and 21-fluoro-118, 17 ex-dihydroxy-6, 16ex-dimethyl-20-oxo - 21 - phenyl-4, 6 - pregnadieno- - [3, 2-c] -pyrazole, and these compounds are separated from each other by Partition chromatography or separated by chromatography on silica gel.
 EMI8.2
 The above-described process of 2'-phenyl-17 ex, 20, 20, 21-bis (methylenedi-1'-phenyl derivative instead of the 21-phenyl derivative, gives the corresponding 1'-phenyl compounds.

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   In all of the processes described above, using the 17 a, 20, 20, 21-bis (methylenedioxy) -2-hydroxymethylene-4, 6-pregnadiene derivative B as the starting material, which in turn is derived from any of the starting materials listed at the beginning of the description is obtained, the corresponding l'-phenyl and 2'-phenyl derivatives.
 EMI9.1
    : droxymethylene-16a-methyl-4, 6-pregnadien-3-one and 5 ml of absolute ethanol are mixed with 49 mg of sodium acetate and then with 95 mg of p-toluene hydrazine hydrochloride. The mixture is refluxed under nitrogen for 45 minutes. The solid precipitate which separates out on cooling is filtered off.



  The filtrate is evaporated to dryness and the residue is mixed with water. The product is filtered, washed with water, dilute acid and again with water until it is neutral. The main component of the solid product is 17 a, 20,20, 21-bis (methylenedioxy) -llss-hydroxy-16a-methyl-2'- (p- - toluyl) -4, 6-pregnadieno- [3, 2- c] pyrazole.



   The product is purified by dissolving 235 mg of the raw material in 30 ml of methanol and stirring with 235 mg of decolorizing charcoal at room temperature. The mixture is filtered and the filtrate is concentrated and allowed to crystallize. 17 a, 20, 20, 21-bis (methylenedioxy) -110-hydroxy-16a-methyl-2'- (p-toluyl) -4, 6-pregnadieno- [3, 2-c] pyrazole is obtained .



   According to another method, a mixture of 1 (p-toluyl) and 2'- (p-toluyl) - - 17 a, 20, 20, 21-bis (methylenedioxy) -118 -hydroxy-4, 6-pregnadieno- [3, 2-c] -pyrazole produced, in-
 EMI9.2
 then reacted with p-toluylhydrazine according to the process of Example 2, using p-toluylhydrazine instead of phenylhydrazine in the second process stage. The components of the mixture are separated from one another by chromatography.



   13 mg of 21- (p-toluyl) -17 a, 20, 20, 21-bis- (methylenedioxy) -11ss-hydroxy-16α-methyl-4,6-pregna- dieno- [3, 2-c] - pyrazole are heated for 30 minutes on the steam bath with 1 ml of 60% aqueous formic acid. The mixture is evaporated to dryness, water is added and the product is filtered off. Man
 EMI9.3
 Left to stand at room temperature overnight. The solvents are removed in vacuo, whereupon water is added and the 118, 17a, 21-trihydroxy-16α-methyl-20-oxo-2 '- (p-toluyl) -4,6-pregna-dieno- [3, 2-c] pyrazole-21-acetate was filtered off.



   After drying, the compound is dissolved in methylene chloride, whereupon a few drops of 2.5N hydrochloric acid are added and the mixture is evaporated to dryness. The 118, 17a, 21-trihydroxy-16α-methyl-20-oxo-4,6-pregnadieno- [3,2-c] -pyrazole-21-acetate hydrochloride obtained in this way is soluble in methylene chloride and can form Acetone are recrystallized.



   If, in the above process, instead of hydrochloric acid, an equivalent amount of sulfuric acid, chloric acid, perchloric acid, picric acid or trichloroacetic acid is added, the corresponding sulfate, chlorate, perchlorate, picrate or trichloroacetate is formed.



   According to the method described in detail in Example 2 for the corresponding 21-phenyl derivatives
 EMI9.4
 If droxymethylene-4,6-pregnadiene derivative is used, which was obtained from one of the starting materials listed at the beginning of the description, the corresponding l'- (p-toluyl) and 21- (p-toluyl) derivatives are obtained.
 EMI9.5
 then mixed with 105 mg of p-methoxyphenylhydrazine hydrochloride. The reaction mixture is heated to reflux temperature for 5 minutes, whereupon it takes on a dark color. The product is filtered and used for

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 Evaporated to dryness. The residue is dissolved in ether, filtered and stirred with an equal amount by weight of decolorizing charcoal. The filtrate is concentrated to 1 ml. Then petroleum ether is added and the product is filtered off.

   The main component of the solid substance obtained in this way is bis (methylenedioxy) - - ss-hydroxy-16ct-methyl-21- (p-methoxyphenyl) -4, 6-pregnadieno- [3, 2-e 1-pyrazole.
 EMI10.1
    Another method is to react a mixture of l'- (p-methoxyphenyl) -latter compound with p-methoxyphenylhydrazine according to the method given in Example 2, by using p-methoxyphenylhydrazine in place of the phenylhydrazine in the second stage of the reaction. The components of the mixture are separated from one another by chromatography.



   70 mg 17ce, 20.20, 21-bis- (methylenedioxy) -11ss-hydroxy-16α-methyl-2 '- (p-methoxyphenyl) - -4, 6-pregnadieno- [3, 2-c] -pyrazole are heated with 7 ml of 600 / o aqueous formic acid for 40 min under nitrogen on the steam bath. The mixture is evaporated to dryness, the residue is mixed with water and the product is filtered. The solid body is purified by dissolving it in acetone, adding 50 mg of decolorizing carbon and filtering.

   Then petroleum ether is added and the product is allowed to crystallize. 11ss, 17α, 21-trihydroxy-16α-methyl-20-oxo-2 '- (p-methoxyphenyl) -4,6-pregnadieno- [3, 2-c] pyrazole is obtained.
 EMI10.2
 
17 cx, 21- trihydroxy - 16cx - methyl- 20 - oxo - 21¯ (p - methoxyphenyl) - 4, 6 - pregnadieno-thoxyphenyl) -4, 6-pregnadieno- [3, 2-c] -pyrazole-21- acetate filtered off. After drying, the compound is dissolved in methylene chloride, a few drops of 2.5N hydrochloric acid are added to the solution and that
 EMI10.3
 ss, 17a, 21-trihydroxy-16a-methyl-20-oxo-2'- (p-me-counted starting materials have been prepared, the corresponding l'- (p-methoxyphenyl) and 2'- (p-methoxyphenyl) are obtained ) derivatives.



    Example 5: 111.5 mg of 17α, 20,20,21-bis (methylenedioxy) -11ss-hydroxy-2-hydroxymethylene-16cx-methyl-4, 6-pregnadien-3-one are in 2.5 ml of ethanol suspended and mixed with 24.5 mg of sodium acetate and then with 48.5 mg of p-fluorophenylhydrazine hydrochloride. After displacing the air with nitrogen, the mixture is quickly brought to reflux temperature.

   After 1 hour of heating at reflux temperature, the mixture is evaporated to dryness and the residue is dissolved in ether. treats the ether layer three times with 2.5N hydrochloric acid, then three times with 2.5N sodium hydroxide solution and then with water. The ether layer is dried over magnesium sulfate, filtered and evaporated in vacuo to a residue, the main component of which is 17 a, 20, 20, 21 -Bis- (methylenedioxy) -2'-p-fluorophenyl-11ss-hydroxy-16α-methyl- -4, 6-pregnadieno- [3, 2-c] -pyrazole. The latter compound is obtained by dissolving the reaction mixture in methanol and recrystallizing.



   According to another method, a mixture of 1 '- (p-fluorophenyl) - and 2' - (p-fluorophenyl) - - 17 et, 20, 20, 21-bis (methylenedioxy) -11ss-hydroxy-16? - methyl-4,6-pregnadieno- [3,2-c] -pyrazole prepared as follows: A mixture of 1 g of 17ce, 20.20, 21-bis- (methylenedioxy) -11ss-hydroxy-2-hydroxymethylene 16α-methyl-4,6-pregnadien-3-one, 200 ml of methanol and 200 mg of p-toluenesulfonic acid are heated to reflux temperature and then left to stand for 1 hour at room temperature. The reaction

 <Desc / Clms Page number 11>

 mixture is now diluted with water and extracted with ethyl acetate. The ethyl acetate extract is washed twice with 2N aqueous sodium hydroxide solution and then with water. It is then dried and concentrated in vacuo.

   17 <x, 20, 20, 2l-bis (methylenedioxy) -110-hydroxy-2-methoxymethylene-16a-methyl-4, 6-pregnadien-3-one is obtained by direct crystallization or by chromatography on acid-washed Aluminum oxide and eluting with mixtures of ether and chloroform.



   A mixture of 500 mg of 17ci, 20, 20, 21-bis (methylenedioxy) -118-hydroxy-2-methoxymethylene-160: -methyl-4, 6-pregnadien-3-one, 100 ml of ethanol and 1 ml of p -Fluorophenylhydrazine is heated under nitrogen until solution has occurred and then left under nitrogen at room temperature overnight. After adding 2 ml of acetic acid, the mixture is left to stand for a further 4 hours. The reaction mixture is then diluted with ethyl acetate, washed twice with 2N sulfuric acid, twice with 2.5N sodium hydroxide solution and finally twice with water.

   The ethyl acetate extract is dried, concentrated and chromatographed on aluminum oxide washed with acid. Eluting with benzene gives
 EMI11.1
    (methylenedioxy) -Iss-hydroxy-16a-methyl-ii-fluorophenyl-4,6-pregnadieno [3, 2-c 1-pyrazolophenyl] -4, 6-pregnadieno [3, 2-c] pyrazole.



   70 mg of 17 a, 20, 20, 21-bis (methylenedioxy) -2'- (p-fluorophenyl) -18-hydroxy-16ct-methyl-4, 6-pregnadieno- [3, 2-c] pyrazole Heated for 35 min on the steam bath with 2 ml of 60% formic acid. The excess of the reagent is driven off in a vacuum on the water bath at approx. The residue is washed thoroughly with water and then dried at 80 ° C; Yield 61.1 mg. The raw product is dissolved in 3 ml of pure methanol for spectral analysis. dissolved and allowed to react with 0.5 ml of a 0.1N sodium methylate solution in methanol for 10 min at room temperature. The product is neutralized with acetic acid.

   Then the mixture is evaporated to dryness, washed thoroughly with water, filtered
 EMI11.2
    Dried constant weight. A solution of 100 mg of 11ss, 17α, 21-trihydroxy-2 '- (p-fluorophenyl) -16α-methyl-20-oxo-4, 6-pregnadieno- [3, 2-c] -pyrazole 1 milliequivalent of acetic anhydride is added to 2 ml of pyridine. The mixture is left to stand at room temperature overnight. Then a mixture of ice and water is
 EMI11.3
 layer is 1-3), saturated aqueous bicarbonate solution (until the pH of the aqueous layer is 8) and water (until the aqueous layer is neutral).

   The ethyl acetate solution is with
 EMI11.4
 
17 (x, 21-trihydroxy-2'- (p-fluorophenyl) -16 <x-methyl-20-oxo-4,6-pregnadieno- [3, 2-c] - pyrazole-21-acetate, which is isolated by adding water and filtering.



   If, in the above process, an equivalent amount of another acylating agent is used instead of the acetic anhydride, the corresponding 21-acylate is obtained.



   A solution of 85 mg of 2 '- (p-fluorophenyl) -11ss, 17α, 21-trihydroxy-16α-methyl- - 20-oxo-4, 6-pregnadieno- [3, 2-c] -, cooled to OOC pyrazole in 0.5 ml of pyridine is mixed with 0.015 ml of methanesulfonyl chloride. The resulting mixture is left to stand at OOC for 1 hour. Then water is added, the crystalline precipitate formed is filtered off, washed with water and dried. There is obtained 2 '- (p-fluorophenyl) -11ss-17α, 21-trihydroxy-16α-methyl-20-oxo-4,6-pregnadieno [3,2-c] -pyrazole- - 21-mesylate.



   A solution of 180 mg of 21- (p-fluorophenyl) -11ss, 17α, 21-trihydroxy-16α-methyl-20-oxo-4, 6-pregnadieno [3, 2-c] -pyrazole-21-mesylate 300 mg of sodium iodide are added to 10 ml of acetone. The mixture is heated to reflux temperature for 1 hour and the reaction solution is then cooled to room temperature and diluted with water. The crystalline precipitate is filtered off, washed with water and dried. There is obtained 2 '- (p-fluorophenyl) -11ss, 17α-dihydroxy-21-iodo-16α-methyl-20-oxo-4, 6-pregnadieno [3, 3, 2-c] -pyrazole.



   The 2 '- (p-fluorophenyl) -11ss, 17α-dihydroxy-21-iodo-167alpha; -methyl-20-oxo-4,6-pregnadieno [3,2-c] - - pyrazole is used in a mixture dissolved from 5 ml of water and 5 ml of ethanol. 500 mg of sodium bisulfite are added to the suspension, and the mixture is heated to reflux temperature for 1 hour. The reaction solution is cooled, diluted with water and the crystalline precipitate is filtered off. The product is washed with water, dried and recrystallized from ethyl acetate. 2'- (p-Fluorophenyl) -11ss, 17α-dihydroxy-16α-methyl-20-oxo-4,6-pregnadieno [3,2-c] pyrazole is obtained.

 <Desc / Clms Page number 12>

 
 EMI12.1
 

 <Desc / Clms Page number 13>

 
 EMI13.1
 le of the 21-benzyl derivative, the corresponding l'-benzyl compounds are obtained.



   If other aralkylhydrazines are used in the above processes, e.g. B. phenylethylene hydrazine, instead of benzyl hydrazine, the corresponding 11- and 21-aralkyl derivatives are obtained.



   If one starts with any 17 ct, 20.20, 21-bis (methylenedioxy) -2-hydroxymethylene-4, 6-pregnadiene derivative, which was obtained from one of the starting materials listed at the beginning of the description, in all of the processes described above the corresponding 1'- and 21-aralkyl derivatives.



     Example 7: A mixture of 71.6 mg 17ct, 20.20, 21-bis (methylenedioxy) -9ct-fluoro-2-hydroxymethylene-16ct-methyl-4, 6-pregnadiene-3, ll-dione and 0.02 ml of phenylhydrazine is heated to reflux temperature for 0.97 ml of absolute ethanol. A crystalline product separates out in the heat. The reaction mixture is cooled and filtered. The product is washed with cold methanol and gives the main component 17a, 20, 20, 21-bis- (methylenedioxy) -9ct-fluoro-16ct-methyl-ll-oxo-21-phenyl-4,6-pregnadieno- [ 3, 2-c] pyrazole.



   29.5 mg of this substance are heated for 1 hour and 5 minutes on the steam bath under nitrogen with 12 ml of added aqueous formic acid. The product is evaporated to dryness, mixed with water and that
 EMI13.2
 Addition of 1 g of sodium borohydride to 100 ml of isopropyl alcohol and filtering off the insolubles produced solution. A drop of water is added and the mixture is then left to stand at room temperature overnight. The excess sodium borohydride is decomposed with acid and the residue is washed with water. 17 <x, 20, 20, 21-bis (methylenedioxy) -9ct-fluoro-1lss-hydroxy-16a-methyl-21-phenyl-4, 6-pregnadieno- [3, 2-c] - pyrazole.



     9.4 mg of this compound are heated under nitrogen on the steam bath for 35 min with 6 ml of 60% strength aqueous formic acid. The product is evaporated to dryness and the residue with water
 EMI13.3
 



   17 ct, 21-trihydroxy-16ct-methyl-Example 8: A solution of 100 mg 17ct, 21-dihydroxy-6a, 16ct-dimethyl-4-pregnen- - 3, 11, 20-trione in 2 ml of pyridine is mixed with 1 Milliequivalent acetic anhydride added. The mixture is left to stand at room temperature overnight. Then a mixture of ice and water is added. After standing for 30 minutes, the product is extracted with ethyl acetate. The ethyl acetate extract is successively mixed with water, ice-cold In sulfuric acid (until the pH of the aqueous layer is 1 to 3), saturated aqueous sodium bicarbonate solution (until the pH of the aqueous layer is 8), and water (until the aqueous layer is neutral) washed. The ethyl acetate solution is dried over anhydrous sodium sulfate and the solvent is then distilled off in vacuo at 40.degree.

   17 ct, 21-dihydroxy-6a, 16ct-dimethyl-4-pregnen-3, l1, 20-trione-21-acetate, which is isolated by adding water and filtering, is obtained.



   A mixture of 1 g of 17 ct, 21-dihydroxy-6 ct, 16 ct-dimethyl-4-pregnen-3, 11, 20-trione-21-acetate, 2 g of chloranil, 25 ml of ethyl acetate and 5 ml of acetic acid is for 10 h under nitrogen heated to boiling and then poured into water. The mixture is extracted several times with ethyl acetate, and the combined extracts are washed with cold pigerous sodium hydroxide solution until the alkaline wash liquid is colorless, and finally washed neutral with water. The ethyl acetate solution is dried and the residue is recrystallized from a mixture of acetone and ether. 17 a, 21-dihydroxy-6, 16ct-dimethyl- - 4,6-pregnadieno-3, 11, 20-trione are obtained.



   A suspension, cooled to 5 ° C. in an ice bath, of 25.0 g of 17α, 21-dihydroxy-6,16α-dimethyl-4,6-pregnadiene-3, 11, 20-trione in 1.5 l of alcohol-free chloroform is added constant stirring with 750 ml of cold, concentrated hydrochloric acid and then with 750 ml of formalin (of low methanol content).



  The mixture is removed from the ice bath and stirred for 7 hours at room temperature. The layers are separated and the aqueous phase is extracted twice with chloroform. The combined organic layers are washed twice with 5% sodium bicarbonate solution and twice with saturated saline solution. The solution is then dried over magnesium sulfate and evaporated to dryness under reduced pressure. The residue is stirred with methanol, with a

 <Desc / Clms Page number 14>

 crystalline solid material is obtained. This material does not contain detectable amounts of starting material by paper strip chromatography, but shows ultraviolet absorbing sites nearby
 EMI14.1
 is used without further purification.



   500 mg of 17ct, 20, 20, 21-bis (methylenedioxy) -6, 16ci-dimethyl-4, 6-pregnadiene-3, 11-dione are dissolved in 25 ml of benzene, whereupon 5 ml of benzene are distilled off at normal pressure. The resulting solution is cooled to room temperature and then 0.75 ml of freshly distilled ethyl formate is added. After the air has been displaced by nitrogen, 150 mg of sodium borohydride (as a 57% strength dispersion in mineral oil) are added and the mixture is stirred for 3 hours under nitrogen at room temperature. Then 15 ml of a saturated aqueous solution of monosodium phosphate are added and the product is extracted with ether.

   The ether extracts are extracted with 2N sodium hydroxide solution and the sodium hydroxide extracts are acidified with monosodium phosphate and extracted again with ether. The ether extract becomes dry
 EMI14.2
    20, 20, 21-bis (methylenedioxy) -2-hydroxymethylene, -3,11-don are dissolved in 25 ml of ethanol. 300 mg of phenylhydrazine are added and the mixture is heated to reflux temperature under nitrogen for 1 hour. Then 25 ml of water are added and the product is extracted with 150 ml of ether. The extracts are washed with 2N hydrochloric acid, saturated sodium bicarbonate solution, water and saturated sodium chloride solution, then dried over sodium sulfate and evaporated to dryness. 1.2 g of crude product are obtained.

   When recrystallizing from ether, the main component of the product 17a, 20, 20, 21-bis (methylenedioxy) -6, 16ct-dimethyl-ll-oxo-21-phenyl-4, 6-pregnadieno- [3, 2-c] pyrazole.



   Example 9: Which becomes 17a, 20,20, 21-bis (methylenedioxy) -6, 16K-dimethyl-ll-oxo-2'-phenyl- -4, 6-pregnadieno- [3, 2-c] -pyrazole suspended in a mixture of 12 ml of tetrahydrofuran and 4 ml of water. Then 1 drop of pyridine and 1 drop of dilute sodium hydroxide solution are added. After adding 160 mg of sodium borohydride, the mixture is heated to reflux temperature under nitrogen until the reduction of the 11-carbonyl group is complete. The excess sodium borohydride is decomposed with aqueous acetic acid. The mixture is evaporated to dryness and treated with water
 EMI14.3
 Nyl-4,6-pregnadieno- [3, 2-c] pyrazole are heated for 30 min on the steam bath under nitrogen with 40 ml of piger aqueous formic acid.

   After adding 40 ml of water, the mixture is extracted with 200 ml of chloroform. The chloroform solution is washed with water, saturated sodium bicarbonate solution and again with water, then dried over sodium sulfate and evaporated in vacuo to give 430 mg of crude product. This is dissolved in 60 ml of absolute methanol, whereupon 0.1 equivalent of sodium methylate in methanol is added. The mixture is stirred for 15 min under nitrogen at room temperature, then acidified with acetic acid and the solvent is driven off in vacuo at room temperature. After adding 20 ml of water, the product is extracted with 150 ml of ethyl acetate. The ethyl acetate solution is washed with saturated sodium bicarbonate solution and then with water, dried over sodium sulfate and evaporated to dryness. An amorphous solid is obtained.



   The crude product obtained in this way is dried in a high vacuum and then dissolved in 4 ml of pyridine. 3 ml of acetic anhydride are added to the solution and the mixture is heated on the steam bath for 15 min and then evaporated to dryness in vacuo. 20 ml of water are then added and the product is extracted with 150 ml of ethyl acetate. The extract is washed with saturated sodium bicarbonate solution and with water and dried over sodium sulfate. The solvent is removed in vacuo and the residue is recrystallized from a mixture of ethyl acetate and benzene. 250 mg of 118, 17a, 21-trihydroxy-6, 16a-dimethyl-20-oxo-21-phenyl-4,6-pregnadieno- [3, 2-c] -pyrazole-21-acetate are obtained.



   100 mg of 118, 17a, 21-trihydroxy-6, 16a-dimethyl-l1, 20-dioxo-21-phenyl-4, 6-pregnadieno [3, 2-c] - - pyrazole-21-acetate are in 10 ml Dissolved absolute methanol and the solution was treated with 1.1 equivalent of sodium methylate in methanol. The mixture is stirred for 15 minutes at room temperature under nitrogen. The product is acidified with acetic acid and then evaporated in vacuo at room temperature. The residue is taken up in 100 ml of chloroform, the solution with water, saturated

 <Desc / Clms Page number 15>

 Sodium bicarbonate solution and washed again with water, then dried over sodium sulfate and evaporated to dryness.

   100 mg of llss, 17,21-trihydroxy-6, 16a-dimethyl-20-oxo-21-phenyl-4,6-pregnadieno- [3, 2-c] pyrazole are obtained.
 EMI15.1
    20, 20, 21-bis (methylenedioxy) -6a, 16a-dimethyl-methyl-11, 20-dioxo-2'-phenyl-4, 6-pregnadieno [3, 2-c] pyrazole.



   Example 11: 440 mg of 17a, 21-dihydroxy-6, 16a-dimethyl-ll, 20-dioxo-21-phenyl-4, 6- pregnadieno [3, 2-c] pyrazole are added in a mixture of 3 , 3 ml of dimethylformamide, 10 ml of methanol, 0.32 ml of water, 255 mg of semicarbazide hydrochloride and 140 mg of sodium bicarbonate suspended. The mixture is heated to reflux temperature under nitrogen for 45 minutes and then heated to 45 ° C. for a further hour. It is then cooled and diluted with water. The resulting precipitate is filtered off and washed thoroughly with water. 17 a, 21-dihydroxy-6, 16a-dimethyl-ll, 20-dioxo- - 2'-phenyl-4, 6-pregnadienor 3, 2-c] -pyrazole-20-semicarbazone are obtained.



   Example 12: The 20-semicarbazone according to Example 4 is suspended in a mixture of 12 ml of tetrahydrofuran and 4 ml of water. 1 drop of pyridine and 1 drop of dilute sodium hydroxide solution are added and, after addition of 160 mg of sodium borohydride, the mixture is heated to reflux temperature under nitrogen until the reduction of the carbonyl group is complete. The excess sodium borohydride is decomposed with aqueous acetic acid.

   The mixture is evaporated to dryness and
 EMI15.2
    a, 21-Trihydroxy-6, 16a-dimethyl-20-oxo-21-phenyl-4, 6-preg-Example 13: A mixture of 250 mg llss, 17 (x, 21-Trihydroxy-6, 16a -dimethyl-20-oxo-21-phenyl-4, 6-pregnadieno- [3, 2-c] -pyrazole-20-semicarbazone, 2.5 ml of acetic acid, 2.5 ml of water and 2.5 ml of pyruvic acid is stirred overnight under nitrogen at room temperature. It is then carefully poured into 5% sodium bicarbonate solution and extracted with chloroform. The extracts are washed and evaporated to dryness. 118, 17 a, 21-trihydroxy-6, 16a-dimethyl- 20-oxo-21 - - phenyl-4, 6-pregnadieno- [3, 3, 2-c] -pyrazole, which is recrystallized from a suitable solvent.



   Although in a preferred embodiment of the invention the 20-position keto group is blocked by the formation of the 20-ethylenedioxy derivatives, as described in Examples 14 and 15, other cyclic ketal derivatives can also be used in the process according to the invention in order to reduce the 20-position to protect dige ketone group. In general, the lower alkylenedioxy derivatives in which the hydrocarbon group contains no more than 7 carbon atoms, such as the ethylenedioxy, trimethylenedioxy, propylenedioxy and butylenedioxy derivatives, have proven to be most suitable. However, instead of a lower alkylenedioxy substituent to block or protect the keto substituent for this purpose, other derivatives which are easily hydrolyzable to keto derivatives can be used, e.g.

   B. an enol ether monothioketal or a dithioketal derivative.



   Example 14: A mixture of 1.0 g of 17α, 21-dihydroxy-6,16α-dimethyl-11,20-dioxo-2'-phenyl-4, 6-pregnadieno- [3, 2-c] - Pyrazole, 14 ml of ethylene glycol and 70 mg of p-toluenesulphonic acid in 200 ml of benzene are heated to reflux temperature for 5-10 hours using a water trap. Excess sodium bicarbonate solution is then added to neutralize the acid. The mixture is cooled, water is added and the mixture is extracted with ethyl acetate. The extracts are washed with water, evaporated to dryness and chromatographed on silica gel in order to obtain the 20-ethylenedioxy derivative.



   The fraction which gives a negative tetrazolium test is suspended in a mixture of 12 ml of tetrahydrofuran and 4 ml of water. 1 drop of pyridine and 1 drop of dilute sodium hydroxide solution are added and, after addition of 160 mg of sodium borohydride, the mixture is heated to reflux temperature under nitrogen until the reduction of the 11-carbonyl group is complete. The excess Na-
 EMI15.3
 

 <Desc / Clms Page number 16>

 ren is cleaned.



   Example 16: A solution of 2 g llss, 17a-21-trihydroxy-6, 16c <-dimethyl-4, 6-pregnadiene--3, 20-dione-21-acetate in 100 ml of dry pyridine is mixed with 23 ml of dry formic acid ethyl ester and then treated with a solution of sodium hydride in 100 ml of pyridine under nitrogen. The reaction mixture is left to stand at room temperature for 42 hours and then poured into ice water. After adding 218 ml of glacial acetic acid, the resinous product is separated off and dissolved in ether. The ether solution is washed with a solution of 30 g of potassium hydroxide in 1.5 l of water and the aqueous layer is cooled to 50 ° C. and acidified with 6N hydrochloric acid. The precipitate is filtered off and dried over phosphorus pentoxide at 60 ° C. in vacuo.

   One receives 11ss, 17 <x, 21-trihydroxy-2-hydroxymethylene-6, 16 <x-dimethyl- - 4, 6-pregnadiene-3, 20-dione.



   The 11ss, 17ct, 21-trihydroxy-6, 16a-dimethyl-4, 6-pregnadiene-3,20-dione-21-acetate used in Example 1 is produced as follows:
A solution of 100 mg of 11ss, 17α, 21-trihydroxy-6α, 16α-dimethyl-4-pregnen-3,20-dione in 2 ml of pyridine is treated with 1 milliequivalent of acetic anhydride. The mixture is left to stand at room temperature overnight. A mixture of ice and water is then added, the mixture is left to stand for a further 30 minutes and the product is extracted with ethyl acetate.

   The ethyl acetate extract is successively mixed with water, ice-cold 1N sulfuric acid (until the pH of the aqueous layer is 1-3), saturated aqueous sodium bicarbonate solution (until the pH of the aqueous layer is 8) and water (until the aqueous layer is neutral is) washed. The ethyl acetate solution is then dried over anhydrous sodium sulfate, the solvent is distilled off at 40 ° C. under vacuum and the resulting liss, 17 ct, 21-trihydroxy-6ct, 16ct-dimethyl-4-pregnen-3, 20-dione-21-acetate is passed through Adding water and filtering isolated.



     A suspension of 300 mg of 11ss, 17α, 21-trihydroxy-6α, 16α-dimethyl-4-pregnen-3,20-dione-21-acetate, 700 mg of chloranil, 8.3 ml of ethyl acetate and 1.7 ml of acetic acid is heated to reflux temperature under nitrogen for 17 h. The reaction mixture is diluted with 40 ml of ethyl acetate and filtered. The filtrate
 EMI16.1
 washed. The aqueous washing liquids are extracted with 50 ml of ethyl acetate and treated as described above.



   The combined organic phase is dried over sodium sulfate and concentrated in vacuo. The raw material dissolved in benzene is chromatographed on 15 g of acid-washed aluminum oxide. Eluting with a mixture of 2 parts by volume of chloroform and 8 parts by volume of ether is obtained
 EMI16.2
 1 milliequivalent acetic anhydride is added to 2 ml of pyridine. The mixture is left to stand at room temperature overnight and then a mixture of ice and water is added. After standing for 30 minutes, the product is extracted with ethyl acetate. The ethyl acetate extract is successively mixed with water, ice-cold In sulfuric acid (until the pH of the aqueous layer is 1-3), saturated aqueous bicarbonate solution (until the pH of the aqueous layer is 8) and water (until the aqueous layer is neutral) washed.



   The ethyl acetate solution is dried with anhydrous sodium sulfate. Then the solvent is distilled off in vacuo at 40.degree. This gives 11ss, 17ct, 21-trihydroxy-6, 16c <-dimethyl-4, 6-pregnadiene-3, 20-dione-21-acetate, which is isolated by adding water and filtering.
 EMI16.3
    17: methyl-3, 20-dioxo-4, 6-pregnadiene-3, 20-dione in acetic acid is allowed to react with exactly one molar equivalent of phenylhydrazine at room temperature under nitrogen for 1/2 hour. Addition of water gives llss, 17α21-trihydroxy-6,16α-dimethyl-20-oxo-2'-phenyl-4,6-pregnadieno- [3,3,2-c] - -pyrazole, which is isolated and purified by recrystallization.



   Example 18: A mixture of 1 g of 6cx, 16a-dimethyl-4-pregnen-3, 20-dione, 2 g of chloranil, 25 ml of ethyl acetate and 5 ml of acetic acid is heated to boiling under nitrogen for 10 h and then poured into water. The mixture is extracted several times with ethyl acetate, and the combined extracts are washed with cold 10% sodium hydroxide solution until the alkaline washing liquid is colorless, and finally washed neutral with water. The ethyl acetate solution is dried and the residue is recrystallized from a mixture of acetone and ether. 6, 16a-dimethyl-4, 6-pregnadiene-3, 20-dione is obtained.

 <Desc / Clms Page number 17>

 



   Example 19: A spore suspension in water is prepared from an Aspergillus ochraceus slant tube (NRRL405) and used to inoculate a 25 Ornl Erlenmeyer flask containing 50 ml of a medium of the following composition:
 EMI17.1
 
<tb>
<tb> 2 <SEP>% <SEP> technical <SEP> digested <SEP> lactalbumin
<tb> 2 <SEP>% <SEP> glucose
<tb> 0.5% <SEP> corn steep water, <SEP>
<tb>
 
The inoculated flask is incubated on a rotating shaker at 280C for 24 hours. The steroid substrate, namely 6, 16a - dimethyl-4, 6 - pregnadien-3, 20-dione, is used as a solution in dimethylformamide (100 mg / ml) corresponding to a concentration of 1.0 g / l broth (0.5 ml per flask) was added.

   After a further 24 hours of incubation under the same ambient conditions as those of the growth phase, the broth is harvested and the product is isolated.



   50 ml of broth to which 50 mg of 6 ct, 16 ct-dimethyl-4, 6-pregnadiene-3, 20-dione have been added, are extracted three times with 20 ml of ethyl acetate each time after incubation according to the above information. The extract is concentrated to an oil in vacuo, the oil is dissolved in a few drops of chloroform and the solution is chromatographed on paper in a system of "Skelly B" benzene (1: 1) / dimethylformamide-methanol (1: 1). Some unreacted starting material is found on the solvent front.



   325 ml of the broth from the incubation of 480 mg of 6a, 16a-dimethylprogesterone are extracted three times with 325 ml of ethyl acetate each time. The extract is washed with water and concentrated to a small volume in vacuo. In paper chromatography in the system n-hexane-benzene (1: l) / dimethylformamide-methanol (l: l), the expected position for 6, 16a-dimethyl-118-hydroxy-4, 6-pregna- dien-3, 20-dione found.



   The reaction product is dissolved in 20 ml of benzene and the solution is poured through a column containing 50 g of acid-washed aluminum oxide. The column is washed out with benzene and the unreacted starting material is eluted with a mixture of ethyl ether and benzene. Development is continued with a mixture of ethyl ether and ethyl acetate and finally with ethyl acetate alone, which elutes the 6, 16a-dimethyl-11B-hydroxy-4, 6-pregnadiene-3, 20-dione.



    Example 20: A solution of 400 mg of 60, 16a-dimethyl-llss-hydroxy-4, 6-pregnadiene-3, 20-dione in 4 ml of pyridine is added to the complex compound formed by adding 400 mg of chromium trioxide to 4 ml of pyridine. The mixture is swirled until thoroughly mixed and then allowed to stand overnight at room temperature. The reaction mixture is poured into water and the aqueous mixture is extracted with ether and then twice with ethyl acetate. The combined ether and ethyl acetate extracts are washed with dilute aqueous sulfuric acid at OOC and then washed neutral with water. The organic solvent layer is dried, the solvents are evaporated in vacuo and the crystalline residue is purified by recrystallization from a mixture of ethyl acetate and ether.

   6, 16a-dimethyl-4, 6-pregnadiene-3, 11, 20- - trione are obtained.



     Example 21: A solution of 8.2 g (0.025 mol) 6, 16a-dimethyl-4, 6-pregnadiene-3, ll, 20-trione in 125 ml of anhydrous tert. 13.6 ml (0.100 mol) of ethyl oxalate and 20 ml (0.062 mol) of a 3.15N solution of sodium methylate in methanol are added over the course of 15 minutes at a temperature above 50 ° C. with stirring, after which the temperature is reduced to 250 ° C. A heavy precipitate forms immediately. The mixture is stirred for 20 min with exclusion of moisture, after which a solution of 3.06 g of sodium acetate and 3.53 ml of glacial acetic acid in 200 ml of methanol is added.



  The precipitate is redissolved and the solution turns dark brown in color.



   The solution is cooled to 50 ° C. in an ice bath and a solution of 10.8 g (0.0675 mol) of bromine in 108 ml of methanol is added over the course of the next half hour. The amount of bromine to be added is determined by the color of the reaction mixture, which becomes progressively lighter when bromine is added until the solution is colorless. If the color of the bromine persists, no more bromine is added.



   45 ml (0.142 mol) of a 3.15N methanolic solution of sodium methylate are added to this solution over the course of 3 hours, whereupon the mixture initially turns deep orange and then brightens to a cloudy, amber-colored solution. The solution is stirred for 5 h at room temperature. A portion of 0.1 mol is dissolved in a mixture of 50 ml of benzene, 25 ml of methanol and 80 ml of acetic acid,

 <Desc / Clms Page number 18>

 whereupon 2.4 g of zinc dust are added to the solution and the whole is stirred vigorously for 30 minutes. The reaction product is poured into 1500 ml of water, washed and dried.
 EMI18.1
 
22: 1,350 g 6, 16 (x-dimethyl-3, ll-dioxo-4, 6, 17 (20) -pregnatriene-21-carboxylic acid-cooled and treated with 0.96 ml of freshly distilled ethyl formate.

   The air is displaced by nitrogen, whereupon 560 mg sodium hydride (as a 58% dispersion in mineral oil) are added. The system is evacuated again and filled with nitrogen, and the mixture is stirred with a magnetic stirrer at room temperature overnight. The mixture is then poured into an excess of a saturated aqueous solution of monosodium phosphate and the product is extracted four times with benzene. The organic extracts are washed three times with water and dried over sodium sulfate. After the solvent has been driven off, the crude product is dissolved in ether and purified as the sodium salt by extraction with 10% sodium carbonate solution. The aqueous alkaline extracts are acidified with an excess of a saturated aqueous monosodium phosphate solution and extracted with ether and chloroform.

   The combined organic extracts are dried over sodium sulfate and evaporated to dryness. The 2-hydroxymethylene derivative is obtained.



   Example 23: A solution of 1 mmol 6, 16cx-dimethyl-3, l1-dioxo-2-hydroxymethylene
 EMI18.2
 se added with stirring to a mixture of 1.5 g of lithium aluminum hydride and 50 ml of anhydrous ether. When the addition is complete, the mixture is heated to reflux temperature for 1/2 hour and then cooled. Ethyl acetate, then water and finally 250 ml of methylene chloride are then carefully added to the mixture with stirring. The mixture is centrifuged and the organic layer is dried. After distilling off the solvent, 118, 21-dihydroxy-6, 16cx-dimethyl-21-phenyl-4, 6, 17 (20) - - pregnatrieno- [3, 2-c] -pyrazole is obtained.



   Example 25: 0.5g llss, 21-dihydroxy-6, 16cx -dimethyl-21-phenyl-4, 6, 17 (20) -pregnatrieno- [3, 2-c] -pyrazole are mixed with 5 overnight at room temperature ml of pyridine and 2 ml of acetic anhydride reacted. The solvents are removed and the product is removed after the addition of water.
 EMI18.3
   21-dihydroxy-6,16of-dimethyl-21-phenyl-4, 6, 17 (20) -pregnatrieno- [3, 2-c] -pyrazole-21-acetate.



   Example 26: A mixture of 0.00033 moles of 11ss, 21-dihydroxy-6,16α-dimethyl-2'-phenyl-
 EMI18.4
 
4, 6, 17 (20) -pregnatrieno- [3, 2-c] -pyrazole-21-acetate0, 26 ml of a 2.6 molar solution of hydrogen peroxide in tert. Butyl alcohol and 0.3 ml of a mixture of 1, 0 g of osmium tetroxide in 100 ml of tert. Butyl alcohol added. A further 0.5 ml of the osmium tetroxide solution are then added in the course of 30 hours. After a further 84 hours, water and methylene chloride are added. The organic solvents are driven off in vacuo. The product is extracted with methylene chloride and the extract is evaporated to dryness. The residue is dissolved in 5 ml of methanol, 1 ml of aqueous sodium bisulfite solution is added and the mixture is heated to 100 ° C. for 1/2 hour.



  Extraction with methylene chloride gives 118,17α, 21-trihydroxy-6,16α-dimethyl-20-oxo- - 2'-phenyl-4,6-pregnadieno- [3,2-c] -pyrazole-21- acetate.



   500 mg of the above compound are dissolved in 2.4 ml of pure methanol and allowed to react for 10 minutes under nitrogen with 0.9 ml of a 1. 33N solution of sodium methylate in methanol at room temperature. The alcoholate is neutralized with acetic acid and the mixture is evaporated to dryness and the residue is rinsed with n-hexane. The residue is washed with water, filtered and dried to constant weight. This gives llss, 17 (x, 21-trihydroxy-6, 16a-dimethyl-20-oxo-2'-phenyl- - 4, 6-pregnadieno [3, 2-c] pyrazole.



   Example 27: A mixture of Ig 11ss, 17α, 21-trihydroxy-6α, 16α-dimethyl-20-oxo-2'-phenyl-5α-prepregnano- [3,2-c] -pyrazole- 21-acetate, 2g chloranil, 25 ml ethyl acetate and 5 ml acetic acid are heated to boiling under nitrogen for 10 h and then poured into water. The mixture is extracted several times with ethyl acetate and the combined extracts are washed with cold 10% sodium hydroxide solution until the alkaline washing liquid is colorless, and finally washed neutral with water.



  The ethyl acetate solution is dried and the residue is recrystallized from a mixture of acetone and ether. 118, 17cx, 21-trihydroxy-6, 16a-dimethyl-20-oxo-2'-phenyl-4, 6-pregna-

 <Desc / Clms Page number 19>

 
 EMI19.1
    2-c] -pyrazol.-6ss, 16a-dimethyl-20-oxo-21-phenyl-5a-pregnano- [3, 2-c j-pyrazole-21-acetate, the same product is obtained.



   The 11ct, 17 ct, 21-trihydroxy- 6ct, 16ct-dimethyl- used as starting material in the above reaction
 EMI19.2
 
20-oxo-21-phenyl-5ct-pregnano- [3, 2-c] -pyrazole-21-acetate become:
50 g of 11B, 17 ct, 21-trihydroxy-6 ct, 16 ct-dimethyl-4-pregnen-3-one are dissolved in 350 ml of methanol and 2 - 5 l of benzene on the steam bath. The steroid is reduced to BASSO with 45.0 g of 5% palladium at room temperature at 2.1 kg / cm2 pressure over the course of 21 hours. The product is filtered and washed three times with a hot mixture of benzene and methanol. The filtrates and washings are concentrated in vacuo on the steam bath. The residue is rinsed twice with acetone. The residue is dissolved in 2.11 acetone and the solution is concentrated to 250 ml.

   It is then left to stand at 50 ° C. overnight, during which process crystallization occurs. The product is a mixture of
 EMI19.3
 tion can be broken down into its components.



   The further procedure is explained on a 5ct pregnane. However, a 5ss-pregnane or a mixture of 5α-pregnane and 5ss-pregnane can be used in the same way. It is also possible to use 68-methyl derivatives of 5ct or 5B pregnan.
 EMI19.4
 
0 g of 11 B, 17 ct, 21-trihydroxy-6 ct, 16 ct-dimethyl-5 ct-pregnane-3, 20-dione are suspended in 735 ml of chloride, and the suspension is poured into a cold mixture of 240 ml of concentrated hydrochloric acid and 240 ml of formaldehyde (of low methanol content), keeping the temperature at 50C. After standing overnight, the layers are separated and the aqueous layer is extracted with 40 ml of methylene chloride.

   The organic layer is washed with water and then three times with 50 ml of sodium bicarbonate solution each time. The organic layer is then dried over magnesium sulfate, filtered and evaporated to dryness in vacuo. The residue is acidified
 EMI19.5
 the dissolved in 23 ml of dry, hot benzene, and the solution is cooled to room temperature and treated with 0.96 ml of freshly distilled ethyl formate. After displacing the air with nitrogen, 560 mg of sodium hydride (as a 58% dispersion in mineral oil) are added. The system is evacuated again and filled with nitrogen and the mixture is stirred overnight with a magnetic stirrer at room temperature. Then the mixture is poured into an excess of a saturated aqueous solution of monosodium phosphate and the product is extracted four times with benzene.

   The organic extracts are washed three times with water and dried over sodium sulfate. After the solvent has been driven off, the crude product is dissolved in ether and purified as the sodium salt by extraction with 10% sodium carbonate solution. The aqueous alkaline extracts are acidified again with an excess of saturated aqueous monosodium phosphate solution and extracted with ether and chloroform. The combined organic extracts are dried over sodium sulfate and evaporated to dryness. 17α, 20,20,21-bis (methylenedioxy) -11ss-hydroxy-2-hydroxymethylene-6α, 16α-dimethyl-5α-pregnan-3-one is obtained.
 EMI19.6
    Mixture of 2'-phenyl-5? -Pregnano- [3,2-c] -pyrazole in 2 ml of pyridine is treated with 1 milliequivalent of acetic anhydride.

   The mixture is left to stand overnight at room temperature and then a mixture of ice and water is added. After standing for 30 minutes, the product is extracted with ethyl acetate. The ethyl acetate extract is successively mixed with water, ice-cold In sulfuric acid (until the pH of the aqueous layer is 1-3), saturated aqueous sodium bicarbonate solution (up to

 <Desc / Clms Page number 20>

 the pH of the aqueous layer is 8) and water (until the aqueous layer is neutral). The ethyl acetate solution is dried with anhydrous sodium sulfate. Then the solvent is distilled off at 400c in vacuo.

   17a, 20, 20, 21-bis (methylenedioxy) -11 8-hydroxy- 6a are obtained. 16a-dimethyl-21-phenyl-5a-pregnano- [3, 2-c] -pyrazole-21-acetate, which is isolated by adding water and filtering.



   Example 28: A mixture of 2 g of 11B, 17a, 21-trihydroxy-6, 16a-dimethyl-4, 6-pregnadiene- 3, 20-dione in 4 ml of dimethylformamide and 15 ml of 2,2-dimethoxypropane is used for 8 hours 10 mg of p-toluenesulfonic acid heated. The mixture is concentrated to remove the excess volatile reactants and poured into a mixture of methylene chloride and saturated saline solution. The organic layer is washed twice with saturated saline solution and once with water and evaporated to dryness.

   This gives llss-hydroxy-17 (x, 21-isopropylidenedioxy-6, 16 (x-dimethyl-4,6-pregnadiene-3, 20-dione, which is purified by chromatography on aluminum oxide.
 EMI20.1
 Methyl-4,6-pregnadiene-3, 20-dione can be produced as follows:
A solution of 100 mg of 11ss, 17α21-trihydroxy-6α, 16α-dimethyl-4-pregnen-3,20-dione in 2 ml of pyridine is treated with 1 milliequivalent of acetic anhydride. The mixture is left to stand overnight at room temperature and then a mixture of ice and water is added. After standing for 30 minutes, the product is extracted with ethyl acetate.

   The ethyl acetate extract is successively mixed with water, ice-cold sulfuric acid (until the pH of the aqueous layer is 1 - 3), saturated aqueous sodium bicarbonate solution (until the pH of the aqueous layer is 8), and water (until the aqueous layer is neutral) washed. The ethyl acetate solution is dried with anhydrous sodium sulfate. The solvent is then distilled off at 40 ° C. in vacuo, and 11ss, 17α21-trihydroxy-6α, 16α-dimethyl-4-pregnen-3,20, dione-21-acetate, which is obtained by adding water and filtering is isolated.



   A suspension of 300 mg of 11 ss, 17?, 21-trihydroxy-6?, 16? -Dimethyl-4-pregnen-3,20-dione-21-acetate, 700 mg of chloranil, 8.3 ml of ethyl acetate and 1, 7 ml of acetic acid is heated to reflux temperature under nitrogen for 17 h. The reaction mixture is then diluted with 40 ml of ethyl acetate and filtered.



  The filtrate is washed in succession twice with 20 ml of ice-cold 10% aqueous sodium bisulfite solution each time, three times with 20 ml of ice-cold 5% aqueous potassium hydroxide solution each time and finally several times with 20 ml of ice water each time. The aqueous washing liquids are back-extracted with 50 ml of ethyl acetate and treated as described above. The combined organic phase is dried over sodium sulfate and concentrated in vacuo.



   The raw material dissolved in benzene is chromatographed on 15 g of acid-washed aluminum oxide. Eluting with a mixture of 2 parts by volume of chloroform and 8 parts by volume of ether gives 11ss, 17α21-trihydroxy-6,16α-dimethyl-4,6-pregnadiene-3,20-dione-21-acetate. This product is then mixed with a solution of potassium bicarbonate in aqueous methanol to form 17 (x, 21-trihydro-
 EMI20.2
    16a-dimethyl-4. 6-pregnadiene-3.nadien-3, 20-dione are dissolved in 23 ml of dry hot benzene and the mixture is cooled to room temperature and 0.96 ml of freshly distilled ethyl formate are added. After displacing the air with nitrogen, 560 mg of sodium hydride (as a 580/0 dispersion in mineral oil) are added.

   The system is evacuated again and filled with nitrogen and the mixture is stirred with the aid of a magnetic stirrer at room temperature overnight. The mixture is then poured into excess saturated aqueous monosodium phosphate solution and the product is extracted four times with benzene. The organic extracts are washed three times with water and dried over sodium sulfate. After the solvent has been driven off, the crude product is dissolved in ether and purified as the sodium salt by extraction with 10% sodium carbonate solution. The aqueous alkaline extracts are acidified again with an excess of saturated aqueous monosodium phosphate solution and extracted with ether and chloroform.

   The combined organic extracts are dried over sodium sulfate and dried to
 EMI20.3
 methyl-4,6-pregnadiene-3, 20-dione.



   Example 30: A solution of 1 mmol of 11ss-hydroxy-2-hydroxymethylene-17α, 21-isopropylidenedioxy-6,16α-dimethyl-4,6-pregnadiene-3,20-dione in 5 ml of glacial acetic acid becomes 30 min reacted at room temperature with 1 mmol phenylhydrazine. The mixture is poured into water and the 11B-hydroxy-17a ,, 21-isopropylidenedioxy-6-16a-dimethyl-20-oxo-21-phenyl-4 6-pregnadieno- [3, 2-c] -pyrazole isolated by filtering off.

 <Desc / Clms Page number 21>

 



     Example 31: 720 mg of 110-hydroxy-17ct, 21-isopropylidenedioxy-6, 16a-dimethyl-20-oxo-2'-phenyl-4, 6-pregnadieno [3, 2-c] -pyrazole are added for 1 h heated under nitrogen on a steam bath with 24 μl 6010figer aqueous formic acid. The excess reagent is removed in vacuo using a water
 EMI21.1
 are dissolved in 2.4 ml of pure methanol and allowed to react for 10 min at room temperature under nitrogen with 0.9 ml of a 1.33N solution of sodium methylate in methanol. The alcoholate is neutralized with acetic acid and the mixture is then evaporated to dryness and rinsed with n-hexane. The residue is washed with water, filtered and dried to constant weight.

   118, 17 a, 21-trihydroxy-6, 16a-dimethyl-20-oxo-21-phenyl-4, 6-pregnadieno [3, 2-c] pyrazole are obtained.



     Example 32; A mixture of 2g 11ss, 17a, 21-trihydroxy-6,16a-dimethyl-4,6-pregnadiene- 3, 20-dione, 5 ml benzene and 15 ml methyl orthoformate is distilled in the presence of 10 mg p-toluenesulfonic acid, whereby the crude cyclic 17 a, 21-methyl orthoester of the steroid is formed, which has the following formula:
 EMI21.2
   Example 33: 1.350 g of the above crude 17K, 21-orthoester are dissolved in 23 ml of dry hot benzene and the solution is cooled to room temperature and 0.96 ml of freshly distilled ethyl formate are added. After displacing the air with nitrogen, 560 mg of sodium hydride (as a 5-8% igue dispersion in mineral oil) are added.

   The system is again evacuated and filled with nitrogen, and the mixture is stirred with the aid of a magnetic stirrer at room temperature overnight. The mixture is then poured into an excess of saturated aqueous monosodium phosphate solution and the product is extracted four times with benzene. The organic extracts are washed three times with water and dried over sodium sulfate. After the solvent has been driven off, the crude product is dissolved in ether and purified as the sodium salt by extraction with sodium carbonate solution. The aqueous alkaline extracts are acidified again with an excess of saturated aqueous monosodium phosphate solution and extracted with ether and chloroform.

   The combined organic extracts are dried over sodium sulfate and evaporated to dryness, giving the corresponding 2-hydroxymethylene derivative.



   Example 34: A solution of 1 mmol of the above 2-hydroxymethylene derivative in 5 ml of glacial acetic acid is reacted with 1 mmol of phenylhydrazine at room temperature for 30 minutes. The mixture is then poured into water and the product filtered off. The N-phenylpyrazole compound of the formula is obtained
 EMI21.3
 

 <Desc / Clms Page number 22>

 
 EMI22.1
 is heated for 10 min with a few drops of hydrochloric acid. The mixture is poured into ice water and the 11 17ct, 21-trihydroxy-6, 16a-dimethyl-20-oxo-21-phenyl-4, 6-pregnadieno [3, 2-c] pyrazole formed is filtered off.



    Example 36: 500 mg of 110, 17a-dihydroxy-6, 16 <x-methyl-4, 6-pregnadiene-3, 20-dione-21-dihydroxy phosphate are dissolved in 25 ml of benzene, whereupon 5 ml of benzene under normal pressure are distilled off. The solution thus obtained is cooled to room temperature and treated with 0.75 ml of freshly distilled ethyl formate. After displacing the air with nitrogen, 150 mg of sodium
 EMI22.2
 dried and the solvent was distilled off at 40 ° C. in vacuo. This gives 11 ss, 17 cx, 21-trihydroxy-6a, 16cc-dimethyl-4-pregnen-3, 20-dione-21-acetate, which is isolated by adding water and filtering off.



    A suspension of 300 mg 118, 17cx, 21-trihydroxy-6cx, 16cx-dimethyl-4-pregnen-3, 20-dione-21-acetate, 700 mg chloranil, 8.3 ml ethyl acetate and 1.7 ml acetic acid is 17 heated to reflux temperature under nitrogen h. The reaction mixture is then diluted with 40 ml of ethyl acetate and filtered. The filtrate is washed in succession twice with 20 ml of ice-cold 10% aqueous sodium bisulfite solution each time, three times with 20 ml of ice-cold 5% aqueous potassium hydroxide solution each time and finally several times with 20 ml of ice water each time. The aqueous washing liquids are extracted with 50 ml of ethyl acetate and treated as described above. The combined organic phase is dried over sodium sulfate and concentrated in vacuo.



   The crude material dissolved in benzene is chromatographed on 15 g of acid-washed aluminum oxide. By eluting with a mixture of 2 parts by volume of chloroform and 8 parts by volume of ether, 118,17α, 21-trihydroxy-6,16α-dimethyl-4,6-pregndiene-3,20-dione-21-acetate is obtained. The product is then mixed with a solution of potassium bicarbonate in aqueous methanol to give 118, 17 cx, 21-trihydro-
 EMI22.3
 Water washed and dried. 11 17 <x, 21-trihydroxy-6, 16a-dimethyl-4, 6-pregnadiene- 3, 20-dione-21-mesylate are obtained.



   180 mg of 11B, 17 a, 21-trihydroxy-6, 16ct-dimethyl-4, 6-pregnadiene-3,20-dione-21-mesylate, dissolved in 10 ml of acetone, are mixed with 300 mg of sodium iodide. The mixture is heated to reflux temperature for 1 hour and the reaction solution is cooled to room temperature and diluted with water. The precipitate is filtered off, washed with water and dried. It consists of 110, 17 a "dihydroxy-21-iodo- -6,16a-dimethyl-4, 6-pregnadiene-3,20-dione.



   Monosilver phosphate is produced by reacting 32 g of trisilver phosphate with 10 ml of 100% phosphoric acid with thorough mixing in a 1 l three-necked round bottom flask. The monosilver phosphate is washed twice with diethyl ether and the ether is decanted to remove some of the phosphoric acid. The monosilver phosphate is then covered with a layer of 200 ml of acetonitrile and the mixture is heated to reflux temperature. Then 20 g of 11 17? -Dihydroxy-21-iodo-6,16? -Dimethyl- -4,6-pregnadiene-3,20-dione are added and the mixture is refluxed for 75 minutes with stirring in a nitrogen atmosphere. The reaction mixture is cooled to room temperature over the course of 1 hour.

   Then 200 g of ice water are added, and the acetonitrile is driven off at a temperature below 250 ° C. in vacuo. The pH of the aqueous suspension obtained becomes

 <Desc / Clms Page number 23>

 adjusted to 6.4 by adding 23 ml of saturated aqueous sodium carbonate solution. The precipitate that forms is filtered off and washed with water until no more ultraviolet-absorbing material can be found in the washing water. The filtrate and the washing water are combined and subjected to freeze-drying to separate the solid material from the water. The solid is stirred with a total of 770 ml of methanol in seven portions.



  The methanol insoluble is filtered off. The filtrate is then concentrated in vacuo to 200 ml and poured through a column charged with 60 g of a cation exchange resin ("IR-120") in the hydrogen ion form. The column is washed out with methanol until the washing liquid no longer contains any ultraviolet absorbing material. The combined eluates and washing liquids are concentrated to 15 ml and mixed with 150 ml of ether. The precipitate that forms is filtered off, washed with ether and dried in a desiccator for 16 h. 118,17a-dihydroxy-6, 16a-dimethyl- 4, 6-pregnadiene-3, 20-dione-21-dihydrogen phosphate are obtained.
 EMI23.1
 the mixture is heated to reflux temperature under nitrogen for 1 h. Then the ethanol is driven off.

   After adding 25 ml of water to the residue, the product is filtered, freed from unreacted phenylhydrazine by washing with hydrochloric acid and then washed with water. 11 ss, 17 (x, 21-trihydroxy-6, 16ci-dimethyl-20-oxo-21-phenyl-4,6-pregnadiene [3, 2-c] -pyrazole-21-phosphate are obtained.
 EMI23.2
 Worthington Corporation) the mixture is incubated at 37 ° C. for 3 hours. The 118, 17 a, 21-trihydroxy- - 6, 16a-dimethyl-20-oxo-21-phenyl-4, 6-pregnadieno- [3, 2-c] -pyrazole is then isolated by extraction with methylene chloride and by recrystallization cleaned.



   Example 39: A mixture of 100 mg 11ss, 17ct, 21-trihydroxy-6ct, 16a-dimethyl-20-oxo- - 2'-phenyl-4-pregneno- [3, 2-c] -pyrazole-21-acetate, 5 ml of dioxane, 50 mg of dicyandichlorobenzoquinone and 10 mg of p-toluenesulfonic acid are heated to reflux temperature under nitrogen overnight. The product is extracted with ethyl acetate, washed with sodium hydroxide and then with water and evaporated to dryness. This gives llss, 17a-21-trihydroxy-6, 16-x-dimethyl-20-oxo-2'-phenyl-4,6-pregnadieno- [3, -pyrazole.



   The above procedure approaches 11ss, 17α, 21-trihydroxy-6ss, 16α-dimethyl-20-oxo-21-phenyl-4-pregneno- [3, 2-c] -pyrazole-21-acetate Instead of the llss, 17e <, 21-trihydroxy-6ct, 16a-dimethyl-20-oxo-21-phenyl-4-pregneno [3, 2-c] pyrazole-21-acetate, the same product is obtained .



   Example 40: A mixture of 1g 11ss, 17α, 21-trihydroxy-6α, -16α-dimethyl-20-oxo-2'-phenyl-4-pregneno- [3, 2-c] -pyrazole- 21-acetate, 2g chloranil, 25 ml ethyl acetate and 5 ml acetic acid are heated to boiling under nitrogen for 10 h and then poured into water. The mixture is extracted several times with ethyl acetate, and the combined extracts are washed with cold pigerous sodium hydroxide solution until the alkaline washing liquid is colorless, and finally washed neutral with water. The ethyl acetate solution is dried and the residue is recrystallized from a mixture of acetone and ether. One obtains llss, 17a, 21-trihydroxy-6, 16α-dimethyl-20-oxo-2'-phenyl-4,6-pregnadieno- [3,2-c] - - pyrazole.
 EMI23.3
 duct



   The 11 a, 17 a, 21-trihydroxy-66a, 16a (x-dimethyl-20-oxo-2'-phenyl-4-pregneno- [3, 2-c] pyrazole used as starting material in the above reaction -21-acetate can be produced as follows:
12, 0g 11ss, 17α, 21-trihydroxy-6α, 16α-dimethyl-4-pregnen-3,20-dione are suspended in 735 ml of methylene chloride, and the suspension is poured into a cold mixture of 240 ml of concentrated hydrochloric acid and 240 ml of formaldehyde (with low methanol content) poured, the temperature being kept at 50C. After standing overnight, the layers are separated from each other. and

 <Desc / Clms Page number 24>

 the aqueous layer is extracted with 40 ml of methylene chloride. The organic layer is washed with water and then three times with 50 ml of sodium bicarbonate solution each time.

   The organic layer is then dried over magnesium sulfate, filtered and evaporated to dryness in vacuo. The residue is chromatographed on acid-washed alumina to give 17ca, 20, 20, 21-bis (methylenedioxy) -11ss-hydroxy-6α, 16α-dimethyl-4-pregnen-3-one.



   1,350 g of 17α, 20,20,21-bis (methylenedioxy) -11ss-hydroxy-6α, 16α-dimethyl-4-pregnen-3-one are dissolved in 23 ml of dry hot benzene, and the solution is cooled to room temperature and treated with 0.96 ml of freshly distilled ethyl formate. After displacing the air with nitrogen, 560 mg of sodium hydride (as a 58% dispersion in mineral oil) are added. The system is again evacuated and filled with nitrogen, and the mixture is stirred with the aid of a magnetic stirrer at room temperature overnight. The mixture is then poured into excess saturated aqueous monosodium phosphate solution and the product is extracted four times with benzene. The organic extracts are washed three times with water and dried over sodium sulfate.

   After removing the solvent, the crude product is dissolved in ether and purified as the sodium salt by extraction with 1 oily sodium carbonate solution. The aqueous alkaline extracts are acidified again with an excess of a saturated aqueous monosodium phosphate solution and extracted with ether and chloroform.



  The combined organic extracts are dried over sodium sulfate and evaporated to dryness giving 17α, 20,20,21-bis- (methylenedioxy) -11ss-hydroxy-2-hydroxymethylene-6α, 16α-dimethyl- -4-precipitation -3-one receives.



   A mixture of 71.6mg 17a. 20. 20, 21-bis (methylenedioxy) -llss-hydroxy-2-hydroxymethylene- 6 <x, 16 <x -dimethyl-4-pregnen-3-one and 0.02 ml of phenylhydrazine is dissolved in 0.97 ml absolute ethanol heated to reflux temperature for 1 h. The reaction mixture is cooled and filtered. The product will
 EMI24.1
    ct, 20, 20, 21-bis (methylenedioxy) -11 ß-hydroxy-6 ct, nyl-4-pregneno - [3, 2-c] -pyrazole in 2 ml of pyridine is mixed with 1 milliequivalent of acetic anhydride. The mixture is left to stand overnight at room temperature and then a mixture of ice and water is added. After standing for 30 minutes, the product is extracted with ethyl acetate.

   The ethyl acetate extract is successively mixed with water, ice-cold In sulfuric acid (until the pH of the aqueous layer is 1 - 3), saturated aqueous sodium bicarbonate solution (until the pH of the aqueous layer is 8) and water (until the aqueous layer is neutral) washed. The ethyl acetate solution is dried over anhydrous sodium sulfate and the solvent is distilled off at 40 ° C. in vacuo. 17α, 20,20,21-bis (methylenedioxy) -11ss-hydroxy-6α, 16α-dimethyl--2'-phenyl-4-pregneno- [3,2-c] -pyrazole-21 is obtained acetate, which is isolated by adding water and filtering off.
 EMI24.2
 

** WARNING ** End of DESC field may overlap beginning of CLMS **.

 

Claims (1)

WeisePATENT CLAIMS: 1. Process for the preparation of new steroid compounds of the pregnan series of the general formula EMI24.3 in which Rl is a β-hydroxy or keto radical or an β-halogen radical, the latter only if X is halogen, Riz is fluorine, hydrogen or a methyl group, R is a hydrogen atom, a <Desc / Clms Page number 25> a-methyl, ss-methyl or methylene group, R4 a hydroxyl group, an acyloxy group containing 1 - 10 carbon atoms or a phosphoryloxy group, R5 an alkyl, cycloalkyl, aralkyl or aryl EMI25.1 EMI25.2 EMI25.3 and- (methylenedioxy) can be linked to an alkylhydrazine, cycloalkylhydrazine, aralkylhydrazine, Arylhydrazine or a salt of such a compound to a compound of the formula EMI25.4 in which R 1, R, Rg, Rs and X have the above meaning and blocking groups can be present in C-17, C-20 and C-21, and any blocking groups present in C-17, C-20 and C-21 removed, and optionally a II-keto group to a hydroxy group before removing the blocking groups in C-17, C-20 and C-21 with a hydrolyzing agent, such as an aqueous solution of an organic acid, and optionally the 21-hydroxy compound in one 21-org. Acylate, in which the acyl group contains 1 - 10 carbon atoms, or a 21-phosphate converts. 2. The method according to claim l, characterized by the use of a starting compound of the formula EMI25.5 <Desc / Clms Page number 26> in which Rl, R, R3 and X have the meaning according to claim 1. 3. The method according to claim l, characterized by using a starting compound of the formula EMI26.1 in which P ", R, Rs and X have the above meanings and R is an alkyl group. EMI26.2 hydrazine converts. 5. The method according to claim 2, characterized in that a starting compound of the formula according to claim 2 is used and the 21-hydroxy compound in a21-org. Acylate or a 21-phosphate converts.