DE1468211B1 - 17alpha, 21-dihydroxy-20-oxo-4,6-pregnadieno- [3,2-c] -pyrazoles and 21-acylates thereof and the pharmacologically acceptable salts of these compounds and a process for their preparation - Google Patents

Description

The invention relates to new 17 ″, 21-dihydroxy-20-oxo-4,6-pregnadieno [3,2-c] pyrazoles and 21-acylates same of the general formulas (I) and (II)

CH ₂ OR ₄

I.

C = O

.OH

R ₅ -N

in which X is a hydrogen or fluorine atom, R _{1 is} a β-hydroxy group, R _{2 is} a hydrogen atom or a methyl group and R _{3 is} a hydrogen atom or an α-methyl group, with the proviso that R ₂ and R ₃ cannot be hydrogen at the same time, R ₄ denotes a hydrogen atom or an acyl group derived from an organic carboxylic acid and R ₅ denotes a phenyl or p-fluorophenyl radical, as well as the pharmacologically acceptable salts of these compounds and a process for their preparation, which is characterized in that a correspondingly substituted III-hydroxy or II-oxo-17 ", 21-dihydroxy-3,20-dioxo-2-hydroxymethylene or -2-alkoxymethylene-4,6-pregnadiene or a corresponding 17.20; 20,21-bismethylenedioxy derivative is reacted with phenylhydrazine or p-fluorophenylhydrazine or hydrochloride, any 11-oxo compound obtained is reduced with sodium borohydride to give the corresponding 11 / ^ -hydroxy compound, a 17.20 present in the compound obtained; Splitting off 20,21-bismethylenedioxy group with the aid of an organic acid and optionally converting the 21-hydroxy compound obtained into a 21-ester of an organic acid and / or into a pharmacologically acceptable salt.

The preparation of which can be used as starting compounds for the process according to the invention 17.20; 20.21 - bismethylenedioxy derivatives can be carried out as described below:

A 17a, 21-dihydroxy-4,6-pregnadiene-3,20-dione of the general formula

CH ₂ OH

in which R ₁ denotes a / I-hydroxy group and R ₂ , R ₃ and X have the meanings given above, the reaction is carried out with aqueous formaldehyde solution in the presence of a strong acid.

The corresponding 17a, 20; 20,21-bis (methylenedioxy) -11 β -hydroxy-4,6-pregnadiene-3-one obtained in this way with chromium trioxide in an anhydrous base,

(N)

like pyridine, to the corresponding 17 ", 20; 20,21 -Bis- (methylenedioxy) -4,6-pregnadiene-3, l 1 -dione is oxidized. However, this connection can also be obtained directly by reacting the 17 ", 21-dihydroxy-4,6-pregnadiene-3,11,20-trione be prepared with formaldehyde solution in the presence of an acid.

Next, by reacting with an alkyl formate and sodium hydride in one inert atmosphere in the 2-position a hydroxymethylene group stirred in. The corresponding J7 ", 20; 20,21-bis (methy! Endioxy) -2-hydroxymethylene-4,6-pregnadien-3-one is obtained in this way the general formula

H ₇ C

HO

in which R ₁ denotes a β-hydroxy or oxo group and R ₂ , R ₃ and X have the meanings given above.

By reacting this compound with a lower alkanol in the presence of an acidic reagent, like p-toluenesulfonic acid, the corresponding one forms 17a, 20; 20,21 -Bis- (methylenedioxy) -2-alkoxymethylene-4,6-pregnadien-3-one.

If the latter compound, a starting compound for the process according to the invention, with a phenylhydrazine or p-fluorophenylhydrazine or hydrochloride is converted, the corresponding 1'-N-substituted (cf. Formula I) or 2'-N-substituted 4,6-pregnadieno- [3,2-c] -pyrazoles (see Formula II). The two isomers can be separated from one another by chromatography.

If the starting material is a 17a, 20; 20,21-bis (methylenedioxy) -2-hydroxymethylene-4,6-pregnadien-3-one is used, is formed in the inventive reaction with p-fluorophenyl hydrazine in general an isomer in predominant amount, while in the implementation of a 17a, 20; 20,21 -Bis- (methy! endioxy) - 2 - alkoxymethylene - 4,6 - pregnadien - 3 - ons with p-fluorophenylhydrazine considerable amounts of both Isomers arise. A mixture of isomers can also be used in the implementation of the monosubstituted

Hydrazine with a 2-hydroxymethylene compound are formed when the same as a result of the manufacturing 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 varying amounts of the 2-alkoxymethylene derivative. A 17 ", 20; 20,21 - bis (methylenedioxy) -1 l / i - hydroxy-4,6-pregnadieno [3,2-c] -pyrazole, which has a 9u-fluoro substituent, is preferably prepared as follows : As described above, the 9 "-fluoro-1 l / i, 17" -21 -trihydroxy- ^ o-pregnadiene-3,20-dione is converted into the corresponding 17a, 20; 20,21 -Bis- (methylenedioxy) - 9a - fluoro-11 β - hydroxy - 4,6 - pregnadien-3-one transferred. This compound is then oxidized to 17 ", 20; 20,21 - bis (methylenedioxy) - 9" - fluoro-4,6-pregnadiene-3, l 1-dione, which with ethyl formate and sodium hydride to form the starting compound 2-hydroxymethylene derivative is implemented. According to the process, the latter is converted into the corresponding one with phenylhydrazine or p-fluorophenylhydrazine or hydrochloride

• N-substituted 17a, 20; 20,21 -Bis- (methylenedioxy) -9fi-fluoro-1 1 -oxo-4,6-pregnadieno- [3,2-c] -pyrazol compound implemented. This connection then becomes the corresponding 17 ", 20; 20,21 -Bis- (methylenedioxy) -9a-fluoro-1 l / i-hydroxy-4,6-pregnadieno [3,2-c] pyrazole compound reduced, 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 let stand overnight.

When treating any of the 17 ", 20; 20.21 - bis (methylenedioxy) - 4,6 - pregnadieno [3,2-c] pyrazole compounds with a dilute organic acid, e.g. B. with 60% aqueous formic acid, the protective 17 ", 20; 20,21-bis (methylenedioxy) group split off, and the corresponding 17 ", 21-dihydroxy-20-oxo-4,6-pregnadieno [3,2-c] pyrazoles are obtained.

Acylating agents suitable for acylating a 21-hydroxysteroid according to the invention are

»For example, acylating agents derived from lower hydrocarbon carboxylic acids, 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, like / ^ - dimethylglutaric anhydride or succinic anhydride. The acylation is carried out in the presence of an organic base such as pyridine.

The 4,6-pregnadieno- [3,2-c] pyrazoles of the invention form salts such as the hydrochloride, sulfate, chlorate, perchlorate, picrate and trichloroacetate, though they are reacted with the corresponding acids. On the way through the crystalline salts, in particular the hydrochlorides, the 4,6-pregnadieno- [3,2-c] -pyrazoles can be isolated.

The 4,6-pregnadieno- [3,2-c] pyrazoles according to the invention have a strong anti-inflammatory properties Activity and are non-toxic. They are especially useful in treating arthritis and the like Diseases effective and can exert their cortisone-like effects in low dosages are administered so that undesirable side effects are suppressed.

The new 4,6-pregnadieno- [3,2-c] -pyrazoles can be processed into pharmaceutical preparations.

example 1

1.19 g of 17 ", 20; 20,21-bis (methylenedioxy) -l / i-hydroxy - 2 - hydroxymethylene - 6.16" - dimethyl - 4,6 - pregnadien-3-one are dissolved in 25 ml of ethanol solved. 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 2η 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, the main constituent 17 ", 20; 20,21 -Bis- (methylenedioxy) - \\ ß- hydroxy - 6,16" - dimethyl - 2 '- phenyl - 4,6 - pregnadieno- [3, 2-c] -pyrazole obtained. Mp = 258-262 C US = -123 (CHCl ₃ ); Ultraviolet: AU ¹ , ¹ , '" ¹¹ 283, 315 πΐμ; f 17 200; 20 800.

430 mg 17 ", 20; 20,21-bis (methylenedioxy) -ll / i-hydroxy-6,16a-dimethyl- T- phenyl-4,6-pregnadieno- [3,2-c] -pyrazole become 30 Heated for minutes on the steam bath under nitrogen with 40 ml of 60% 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 and 0.1 molar equivalent of sodium methylate in methanol is added to the solution. The mixture is stirred for 5 minutes 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 is recrystallized from a mixture of ethyl acetate and benzene. 250 mg II / f, 17a, 21-trihydroxy-6,16a-dimethyl-20-oxo-2'-phenyl-4,6-pregnadieno- [3,2-c] -pyrazole-21-acetate are obtained. Mp. (Double) = 160 to 165 ° C and 229 to 230 ⁰ C; [a]? + 14 ° (CHCl ₃ ); Ultraviolet:) ^ _ax (CH ₃ OH) 283, 315 ηΐμ, t = 15700; 19000.

B e i s ρ i e 1 2

111.5 mg of 17a, 20; 20,21-bis (methylenedioxy) -l 10-hydroxy-2-hydroxymethylene-6,16a-dimethyl -4,6-pregnadien-3-one are suspended in 2.5 ml of ethanol 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 refluxing

it is evaporated to dryness, the residue is dissolved in ether, the ether layer is treated three times with 2.5N hydrochloric acid, then three times with 2.5N sodium hydroxide solution and finally with water. The ether layer is dried over magnesium sulfate, filtered and evaporated in vacuo to a residue, the main component of which is 17c ^ 20; 20,2I - bis (methylenedioxy) -2 '- ρ - fluorophenyl - 1 iß - hydroxy - 6,16α - dimethyl- Is 4,6-pregnadieno [3,2-c] pyrazole. The latter compound is obtained by dissolving the reaction mixture in methanol and crystallizing.

70mg 17a, 20; 20 ^ 1-bis (mcthykndioxy) -2 '- (p-fluoro-

phenylH 1 /? - hydroxy-6,16u-dimethyl-4,6-pregnadieno- [3,2-c] -pyrazole are heated for 35 minutes on the steam bath with 2 ml of 60% formic acid. The excess of the reagent is removed in vacuo on a water bath at about 50 ⁰ C. The residue is washed thoroughly with water and then ^{dried at 80 °} C .; Yield 61.1 mg. The crude product is dissolved in 3 ml of spectral analytically pure methanol and allowed to react with 0.5 ml of a 0.1 N sodium methylate solution in methanol for 10 minutes at room temperature. The product is neutralized with acetic acid. The mixture is then evaporated to dryness, washed thoroughly with water, filtered and dried to constant weight. Il / J, 17o, 21-trihydroxy-2 '- (p-fluorophenyl) -6,16 "-dimethyl-20-oxo-4,6-pregnadieno- [3,2-c] -pyrazole is obtained. Mp 197-203 C; [«] Έ ⁵ -26 ° C (acetone); Ultraviolet: ^ _x (CH ₃ OH) 280, 313 ηΐμ,, 15800, 19300.

Example 3

A mixture of 71.6 mg 17a, 20; 20,21-bis (methylenedioxy) - 9 "- fluoro - 2 - hydroxymethylene - 6.16" - dimethyl-4,6-pregnadiene-3, ll-dione and 0, 02 ml of phenylhydrazine is dissolved in 0.97 ml of absolute ethanol for 1 hour heated to reflux temperature 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 17c ^ 20; 20,21 -Bis- (methylenedioxy) -9a-fluoro-6,16a-dimethyl-11-oxo-2'-phenyl-4,6-pregnadicno- [3 , 2-c] pyrazole.

15 mg 17a ^; 20,21-bis (inethylendioxy) -9a-fluoro-6,16a-dimethyl-11 -oxo ^ '- phenvM.o-pregnadieno- [3,2-c] -pyrazole are: dissolved in methylene chloride, with 0, 03 ml of a reagent prepared by dissolving 0.55 ml of triethylamine in 1.45 ml of isopropyl alcohol and then mixed with 2.5 ml of a solution prepared by adding 1 g of sodium borohydride to 100 ml of isopropyl alcohol and filtering off the insolubles. One 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. 17a, 2O; 2O, 21-bis (methylenedioxy) - 9a - fluorine -110- hydroxy - 6.16 «- dimethyl-2'-phenyl-4 _t 6-pregnadieiK> - [3.2-c] -pyrazole.

9.4 mg of this compound are grounded under nitrogen on the steam bath for 35 minutes with 6 ml of 60% strength heated aqueous formic acid. The product is evaporated to dryness and the residue with Water added. After filtering off, the product obtained is 9 "-Fhior-II / U7o ^ l-trihydroxy-6,16a-dimethyl-20-oxo-2'-phenyl-4,6-pregnadieno- [3.2-c] - pyrazole. Mp (decomposition) 241 to 245 ° C; [«]? -46 ° (CHCl ₃ ); Ultraviolet: Λ Ι ,, ¹ , ¹ / '"283, 3Mm ₁ Jt, ί17700; 20900.

Pharmacological test report

The anti-inflammatory effect of the compounds according to the invention 1 l / f, 17a, 21-trihydroxy-6,16a - dimethyl - 20 - oxo - 2 '- phenyl - 4,6 - pregnadieno- [3,2-c] -pyrazoI-21-acetal (Example 1), Ilftl7a, 21-trihydroxy - 2 '- (p-fluorophenyl) -6,1 6a - dimethyl - 20-oxo-4,6-pregnadieno- [3,2-c] -pyrazol (Example 2) and 9 "- fluorine - II / U7a, 21 - trihydroxy. - 6,16a - dimethyl-20 - oxo - 2 '- phenyl - 4,6 - pregnadieno - [ 3,2-c] pyrazole (Example 3) was compared with that of the known compounds hydrocortisone, triamcinolone, triamcinolone acetonide and dexamethasone by means of the granuloma test.

Description of the granuloma test

A weighed cotton ball was implanted under the skin on each ventral side of young adult male rats. Each steroid came in two or multiple doses administered. The selection

the dosage amounts were based on

* try in which the degree of granuloma (20 to 50%) was comparable to the inhibition, which in rats given a dose of 0.5 to 4.5 mg Hydrocortisone had been observed. The trials were made in rats at the same time, firstly no steroid, secondly hydrocortisone in Standard dosages and thirdly the test compound received. The steroids were given orally once a day Administered for 7 days.

After 7 days the rats were sacrificed; the beads were removed together with the surrounding granuloma, dried at about 90.degree and weighed. From the difference of the granuloma weight in rats given a steroid, and in rats not given a steroid, the percent inhibition of granuloma formation was for each dose of test compound and hydrocortisone standard is calculated. From a graphic Representation of the results, the dosage of the test compound was determined which is required to get the same result as with hydrocortisone. The effectiveness of the test compound is as the weight of hydrocortisone divided by that Weight of test compound, expressed that for Achievement of a given degree of inhibition (20 to 50%) is required.

The superior effectiveness of the compounds according to the invention (Examples 1 to 3) compared to the known compounds can be seen from the table below:

link Hydrocortisone

The compound of example 1 The compound of example 2 The compound of example 3

Triamcinolone Triamcinolone acetonide Dexamethasone

effectiveness 1
551
600
2000
4th
40
170

Claims (1)

Patent claims: 1. 17 «, 21-dihydroxy-2 () - oxo-4,6-pregnadieno [3.2-c] pyrazoles and 21-acylates of the same general formulas I and II CH ₂ OR ₄ (D —Ι R, CH ₂ OR ₄ C = O OH - R ₁ (Π) in which X is a hydrogen or fluorine atom. R, a / i-hydroxy group, R ₂ a hydrogen atom or a methyl group and R _{3 is} a hydrogen atom or a «-methyl group, with the proviso that R ₂ and R ₃ cannot be hydrogen at the same time, R _{4 is} a hydrogen atom or an acyl group derived from an organic carboxylic acid and R _{5 is} a phenyl or p- Fluorophenyl radical means, as well as the pharmacologically acceptable salts of these compounds. 2. ll / 117r /, 21 - trihydroxy - 6,16 «- dimethyl-20-oxo-2'-phenyl-4,6-pregnadieno- [3,2-c] -pyrazole- 21-acetate. 3. II / i.l7 ".21 -Trihydroxy-2 '- (p-fluorophenyl) -6.16" - dimethyl - 20 - oxo - 4.6 - pregnadieno - [3,2-c] pyrazole. is 4. 9 "-Fluor-II / i, 17a, 2I-trihydroxy-oJOM-di- methyl-20-oxo-2'-phenyl-4,6-pregnadieno [3,2-c] pyrazole. 5. Process for the preparation of the compounds according to claim 1, characterized in that one in a manner known per se a correspondingly substituted 1! / ^ - hydroxy or ll-oxo-17 ", 21-dihydroxyO ^ O-dioxo ^ -hydroxymethylene or -2-alkoxymethylene-4,6-pregnadiene or a corresponding 17.20; 20.2I -ismethylene dioxy derivative with phenylhydrazine or p-fluorophenylhydrazine or hydrochloride, an approximately obtained 11-oxo compound with sodium borohydride to give the corresponding 11 / i-Hydroxy compound reduced, one present in the compound obtained 17.20: 20.21 - Bismethylenedioxy group is split off with the aid of an organic acid and, if necessary the obtained 21-hydroxy compound in a 21-lister of an organic acid and or in a converts pharmacologically acceptable salt. 009 517/199