FI84487B - 1-alkyl-androsta-1,4-diene-3,17-diones used as contraceptives and a composition containing them - Google Patents

Abstract

The invention relates to 1-alkyl-androsta-1,4-diene-3,17- diones of the formula I <IMAGE> wherein R1 is a methyl, ethyl, hydroxymethyl, C1 -3 - alkoxymethyl, or a C1 -4 -alkanoyloxymethyl group, R6 is a hydrogen atom or a methyl group, and R7 is a hydrogen atom or a methyl group in the 7α- or 7β- position.

Description

1 84487

1-Alkyl-androsta-1,4-diene-3,17-diones for use in contraception and compositions containing them

Separated from Patent Application No. 842,457.

5

The invention relates to 1-alkyl-androsta-1,4-diene-3,17-diones of general formula I for use in contraception.

15 R6 where

Rx is a methyl, ethyl, hydroxymethyl, C1-4 alkoxymethyl or C1-4 alkanoyloxymethyl group, R6 is a hydrogen atom or a methyl group, and R7 is a hydrogen atom or a methyl group in the 7a or 76 position.

It was found that the new compounds of general formula I inhibit estrogen biosynthesis. This is surprising since the corresponding 1-unsubstituted androsta-1,4-diene-3,17-diones allow a clear increase in estrogen levels.

DK-A-131 786 discloses A4-3-oxo-1α-methyl steroids, while the novel compounds of the formula I are 1-alkyl-androsta-1,4-diene-3,17-diones, which thus have additional compounds compared to the known compounds. 30 double bond, which also changes the etheric environment of the carbon atom in position 1. The compounds known from DK publication 131 786 have a strong anabolic effect and a very significantly reduced androgenic effect. Thus, the novel compounds of the formula I instead inhibit the biosynthesis of estrogens by inhibiting the enzyme aromatase.

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The conversion of androgens to estrogens plays an important role in the biosynthesis of estrogens. This change occurs through a series of reactions, collectively referred to as "aromatization." The aromatase enzyme is a 5-rate enzyme in the conversion of androstenedione and testosterone to estrone and estradiol.

The compounds of the invention were tested for their aromatase inhibitory effect and endocrine pharmacological side effects. The compounds used in the studies were: A: 4-hydroxy-4-androstene-3,17-dione (as a reference in the study of aromatase inhibitory activity, estrogenic activity and in the test for inhibition of testicular function).

B: 1-methyl-androsta-1,4-diene-3,17-dione.

C: 1,7a-Dimethyl-androsta-1,4-diene-3,17-dione.

D: 1,6a-Dimethyl-androsta-1,4-diene-3,17-dione.

1) PMSG test in rats to investigate the aromatase inhibitory effect 20 Investigate estrogen production stimulated by PMSG (Pregnant Mare Serum Gonadotropin) with test substances A, B, C and D. The increased secretion of estradiol by PMSG treatment (comparison) can be reduced by administering an aromatase inhibitor.

.25 20-day-old female rats were pretreated by subcutaneous administration of a total of 7 x 100 I.U. PMSG. 1 hour before and 8 hours after the last PMSG administration, the animals were injected with the test substance. The control animals received vehicle only. 24 hours after the last PMSG administration, the animals were sacrificed. The serum was then radioimmunologically determined for estradiol. The results are expressed as the nanomolar / liter (nmol / l) estradiol concentration and the standard deviation.

35 The significance of the differences obtained compared to the control group was examined using analysis of variance. The intensity of the effect was determined by regression / covariance analysis.

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table 1

Effect of estradiol concentration in peripheral serum-5 in MPSG-pretreated rats__

Anne © Estradiol- Relative effect- mg / animal n concentration 2 x s.c. nmol / l (Λ = 1 »0) 10., ... (0.2 ml PHSG control group strain: ja_, 0 substances) o, i 10 '1.31 + 1.70 A 0.3 10 3, 19+ 1.31 10 15 1.0 10 1.50 + o, 92 3.0 10 1, 52 + 0.7 0.1 10 3.53 + 1.0¾ R 0.3 10 2.92 + 1 , 09. n ZO 1.0 10 1.80 + 1.13 3.0 10 0.78 + 0.181 0.1 10 2.21 + 1.05 25 C ° .3 10 ° '81 ί ° '32 1 6.3 1.0 10 O, '19 + 0.211 3.0 10 0.37 + 0.11 0.1 10 3.73 + 0.89 30 D 0'3 10 2 · 78 + 1.17 e li2 1.0 10 1.78+ 0.67 3.0 101.02 + 0.38 - significant difference compared to PMSG control group oc 03 n - number of animals group colitis 4 84487

Compounds B, C and D according to the invention, as well as reference substance A, cause a dose-dependent decrease in the peripheral serum estradiol concentration in rats, increased by PMSG (Table 1).

5 B and D are then approximately as potent as reference substance A, while C is about 6 times more potent than reference substance A.

2) Uterus / vaginal growth test in mice and rats to investigate the estrogenic effect Rats (200 g) and ovariectomized mice (30 g) in each case, respectively, received the test substance subcutaneously once a day for 5 days. Control animals received vehicle only.

The day after the last treatment, animals 15 were killed. The uterus and vagina were removed immediately thereafter and weighed.

For each group, the mean and standard deviation of organ weight were reported. The significance of the differences obtained compared to the control group was examined by means of analysis of variance.

5 84487

Table 2

Estrogen test in ovariectomized mice 5 _

Dose Body weight (mg) mg / animal / day Uterus Vagina s.c.

10 E2 comparison group 0.00003 126.5 ± 20.5 * 55.9 ± 7.8 3 48.4 ± 8.8 * 16.6 ± 3.3 A 10 64.7 ± 13.1 * 23, 1 ± 4.4 15 3 23.8 ± 6.7 17.7 ± 4.5 B 10 13.7 ± 3.0 10.6 ± 1.5 Oil comparison group - 32.3 ± 9.2 23, 5 ± 6.5 20 * significant difference compared to control group E2 = estradiol 6 84487

Table 3

Oestrogen test in ovariectomized rats 5 Dose Body weight (mg) mg / animal / day Uterus Vagina s.c.

E2 comparison - 0.00001 214.4 ± 25.1 * 102.3 ± 11.8 * 10 group 3 94.3 ± 21.9 52.9 ± 5.7 A 10 131.9121.0 68.1 ± 6.2 15 3 85.0 ± 7.6 55.8 ± 7.4 B 10 87.4 ± 21.1 56.6 ± 9.4 Oil comparison - 94.5 ± 22.8 66, 0 ± 20.5 group 20 _ * significant difference compared to control group E2 = estradiol 7 84487

Compound B of the invention (3 and 10 mg / animal / day, 5 x s.c.) did not show estrogenic activity in rats or mice (Tables 2 and 3). In contrast, reference substance A causes a significant increase in uterine weight at doses of 3 and 10 mg / animal / day 5 (5 x s.c.).

3) Test for the prevention of testicular function in rats to investigate the anti-gonadotropic effect

Male immature rats (30 g) were administered test substance subcutaneously once daily for 12 days. The day after the last treatment, the animals were sacrificed and testicular weights were determined. Control animals received vehicle only.

For comparison, body weights of the prostate, vesicular glands, and adrenal glands were determined.

Body weights were converted to mg per 100 g body weight. Mean and standard deviation were calculated for each group.

The significance of the differences obtained compared to the control group was examined by means of analysis of variance.

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9 84487

Compound B of the invention, at the doses tested (1 and 3 mg / animal / day 12 x sc) in the test for inhibition of testicular function, has no effect on testicular weight (no antigonadotropic effect), but at a dose of 1 mg / animal / day a slight decrease in the weight of the vesicular glands. In contrast, reference substance A causes a significant reduction in the body weights of the testes, vesicular glands and adrenal glands.

In the anabolic / androgenic assay, Compound B of the invention has no effect on the fresh weights of the vesicular glands, prostate, M. levator ani muscle, and adrenal glands.

Experiments 1-3 show that the compounds of the invention are potent aromatase inhibitors that are substantially neutral endocrine pharmacologically.

As aromatase inhibitors, the novel compounds of the general formula I are suitable for inhibiting estrogen biosynthesis.

20 These new compounds are also valuable substances that affect fertility. They can be used as fertility inhibitors to prevent ovulation or egg attachment in women of childbearing potential.

The compounds of formula I may be administered orally or parenterally, for example intraperitoneally, intramuscularly, subcutaneously or percutaneously. The compounds can also be implanted in tissue.

The amount of compound to be administered will vary over a wide range and may cover any effective amount. Depending on the route of administration, the amount of the compound to be administered may be 0.01 to 100 mg / kg of body weight, preferably 0.1 to 20 mg / kg of body weight per day.

Capsules, pills, tablets, granules, etc. are suitable for oral administration. Dosage units may contain, in addition to the active ingredient, a physiologically acceptable carrier such as starch, sugar, sorbitol, gelatin, lubricant, silicic acid, talc, etc. Private dosage units for oral administration may contain, for example, 10 to 100 mg of active ingredient (aromatase inhibitor).

For parenteral administration, the active ingredients may be dissolved or suspended in a physiologically acceptable diluent. Oils are very often used as diluents, with or without a solubilizer, surfactant, suspending or emulsifying agent. Examples of oils used are, for example, olive oil, peanut oil, cottonseed oil, soybean oil, castor oil and sesame oil.

The compounds may also be used in the form of a sustained release injection or implant tablet, which may be formulated so as to provide delayed release of the active ingredient.

The implant tablets may contain as inert substances, for example, biodegradable polymers or synthetic silicones, such as silicone rubber. In addition, the active ingredients may be incorporated into a patch, for example, for percutaneous administration.

The invention therefore also relates to compositions for use in contraception which contain a compound of general formula I.

The compounds of general formula I can be prepared by: a) 17-hydroxysteroid having the general formula

30 va II

il 84487

OH

5f (II> oVy \; "6 wherein R1, R6 and R7 are as defined above, the 17-hydroxy group is oxidized to the 17-oxo group, or

b) A ring-saturated or simply unsaturated steroid of general formula III

3.5 i1 [I

- (III) 20 wherein X is O, H (OH) or H (OAc), wherein Ac is lower acyl having 1 to 4 carbon atoms, Rx, Rt and R, are as defined above and the A4 double bond is optional , dehydrated at the 1,2-, 1,2- and 4,5-positions and the free or liberated 17-25 hydroxy group is oxidized, or

c) A ring-saturated steroid of general formula IV

X

30 R1 I

I I (IV) H! 35 R 6 i 84487 wherein X is O, H (OH) or H (OAc), wherein Ac is a lower acyl group having 1 to 4 carbon atoms and R 1 (R 1 and R 2 are as defined above, dehydrated with 1,2 and 4 , In the 5-position and the free or liberated 17-hydroxy group is oxidized, or 5 d) an alkanoyloxy group in a 1-alkanoyloxymethyl steroid of general formula 1'0 CM20Acl 10 Ia J - 1 (r) R 6 1 to 4 carbon atoms and R 6 and R 7 are as defined above, are hydrolysed and optionally converted to the C 1-3 alkyl ether or C 2-4 alkanoyl ester of the liberated hydroxy group of the 1-hydroxymethyl steroid.

Reactions a) to d) are performed according to known methods of steroid chemistry.

The oxidation according to process variant a) can be carried out in a manner known per se, for example with the aid of chromium-acid reagents (Jones reagent or chromic acid-pyridine) or with pyridinium dichromate or chlorochromate.

According to the 1.2-dehydrogenation method b), it can be carried out according to known methods with dehydrating agents such as selenium dioxide or dichlorodicyanobenzoquinone.

Double bonds at the 1,2- and 4,5-positions can be obtained simultaneously, for example by bromination to 2,4-dibromo-3-ketone and dehydrobromination of the dibromide with lithium or calcium carbonate and lithium bromide in dimethylformamide.

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The 1-hydroxymethyl group liberated in the reaction can then be esterified or etherified, and the liberated 17-hydroxy group can then be oxidized.

The esterification of the 1-hydroxymethyl group in the presence of a 17-hydroxy group is preferably carried out with lead acetate or acetic anhydride in dimethylformamide.

The oxidation of the 17β-hydroxy group is carried out according to process variant a).

In the dehydration according to process variant c), both chemical dehydrating agents and microbiological dehydration are possible. Suitable microorganisms for the formation of 1,2- and 4,5-double bonds are, for example, Schizomycetes microorganisms, such as Arthrobacter simplex (ATCC 6946), Bacillus lentus 15 (ATCC 13805) or Bacillus sphaericus (ATCC 7055).

The hydrolysis of the 1-alkanoyloxymethyl group according to method d) can take place with an inorganic base in an alcoholic solution; possibly the subsequent re-esterification is preferably carried out with the corresponding 20aA acid anhydride in the presence of organic bases.

According to a preferred embodiment, in order to convert the 1-hydroxymethyl steroid of the general formula I into the corresponding 1-alkoxymethyl ethers, the 1-hydroxymethyl steroid is reacted with p-toluenesulfonic acid chloride and pyridine to give 1-tosyloxymethyl steroid followed by alkali alcohol dichloroethyl ester.

The following examples are intended to illustrate the process of the invention in more detail: Example 1 6.01 g of 17S-hydroxy-1-methyl-androsta-1,4-dien-3-one are dissolved in 100 ml of acetone. To this solution is added dropwise at room temperature 22 ml of chromic acid solution (prepared from 6.67 g of CrO3, 6 ml of concentrated H2SO4 and adding 35 ml of water to a volume of 100 ml). The solution is stirred for 1 hour and then precipitated in ice water, the product is filtered off with suction and dried. After recrystallization from acetone / hexane, 5.25 g of 1-methyl-androsta-1,4-diene-3,17-dione with a melting point of 165-166 ° C are obtained.

Example 2 30 g of 1α, 7α-dimethyl-4-androstene-3,17-dione in 700 ml of dioxane are heated with 30 g of dichlorodiacyanobenzoquinone under reflux for 8 hours. After cooling, the insolubles are filtered off with suction, washed with dioxane and the filtrate is concentrated by evaporation in vacuo. The residue is chromatographed on silica gel and recrystallized from acetone / hexane. 18 g of 1,7a-dimethyl-androsta-1,4-diene-3,17-dione with a melting point of 148-151 ° C are obtained.

The following compounds are prepared analogously: 1,78-dimethyl-androsta-1,4-diene-3,17-dione, m.p. 196-179 ° C, 1,178-dimethyl-4-androstene-3.17 from dione, 1,6a-dimethyl-androsta-1,4-diene-3,17-dione, m.p. 234-235 ° C, from 1,6a-dimethyl-4-androstene-3,17-dione 1,1-ethyl-androsta-1,4-diene-3,17-dione with a melting point of 175-176 ° C from 11α-ethyl-4-androstene-3,17-dione.

Example 3 9.1 gla-acetoxymethyl-178-acetoxy-5a-androstan-3-one (melting point 193-194 ° C) [prepared from 3,3-ethylenedioxy-1-methylene-5a-androstan-178-ol (Naturwissen-Schaften 51, (1963) 86), by hydroboration, acetylation and ketal cleavage] is dissolved in 90 ml of glacial acetic acid. To this solution is added dropwise 3.2 ml of bromine in 25 ml of glacial acetic acid and the solution is then stirred for 10 minutes. After precipitation in sodium sulfite-containing ice water, the product is filtered off with suction, washed with water and dried. 16.5 g of crude dibromide are mixed in 100 ml of dimethylformamide with 10 g of lithium bromide and 84487 with 20 g of calcium carbonate for 3.5 hours at a bath temperature of 120 ° C. After cooling, the inorganic salts are filtered off with suction and washed with 150 ml of methanol. To the filtrate is added 6 g of potassium hydroxide in 20 ml of water and stirred for 16 hours at room temperature. The reaction solution is poured into ice water, the product is filtered off with suction and dried. The crude 1-hydroxymethyl-17β-hydroxyandrosta-1,4-dien-3-one (7.5 g) is dissolved in 100 ml of dimethylformamide, 1 g of lead acetate 10 and 15 ml of acetic anhydride are added and the mixture is stirred for 6.5 g. hours at room temperature. After precipitation in water and suction filtration of the product, it is oxidized with chromic acid as described in Example 1. In this case, after recrystallization from acetone / hexane, 2.3 g of 1-acetoxy-15-methyl-androsta-1,4-diene-3,17-dione with a melting point of 121-123 ° C are obtained.

Example 4 2.2 g of 1-acetoxymethyl-androsta-1,4-diene-3,17-dione are mixed in 20 ml of methylene chloride and 20 ml of methanol with 250 mg of potassium hydroxide at 0-5 ° C under an argon atmosphere. under 5 hours. It is then neutralized with acetic acid, concentrated by evaporation and precipitated in water. The product is filtered off with suction, washed with water and dried. After recrystallization from acetone / hexane, 1.7 g of 1-hydroxymethyl-androsta-1,4-diene-3,17-dione with a melting point of 201-202 ° C are obtained.

Example 5 500 mg of 1-hydroxymethyl-androsta-1,4-diene-3,17-30 dione are dissolved in 5 ml of pyridine, the solution is cooled to 0 ° C, 350 mg of p-toluenesulphonyl chloride are added and stirred for 3 hours. at room temperature. After precipitation of the product with water, suction filtration, washing with water and drying, 700 mg of crude tosylate are obtained. This crude product is stirred in 5 ml of toluene with 500 mg ie 84487 of potassium methylate for 5 hours at 20 ° C. It is then diluted with ether, washed with dilute hydrochloric acid and water and concentrated by evaporation in vacuo. After chromatography on silica gel and recrystallization from hexane / acetone, 325 mg of 1-ethoxymethyl-androsta-1,4-diene-3,17-dione with a melting point of 95-98 ° C are obtained.

Claims (9)

1. 1-alkyl-androsta-1,4-diene-3,17-diones of general formula I for use in contraception, in which Rx is methyl, ethyl, hydroxymethyl, A C 1-3 alkoxymethyl or C 1-4 alkanoyloxymethyl group, R 6 is a hydrogen atom or a methyl group, and R 7 is a hydrogen atom or a methyl group in the 7a or 7B position. 2. A compound according to claim 1, which comprises 1-methyl-androsta-1,4-diene-3,17-dione. Compound according to Claim 1, characterized in that it is 1-methylandrosta-1,4-diene-3,17-dione. 3. A compound according to claim 1, which comprises 1,7α-dimethyl-androsta-1,4-diene-3,17-dione or 1,7β-dimethyl-androsta-1,4-diene-3,17 -dione. 4. A compound according to claim 1, which is 1,6a-dimethyl-androsta-1,4-diene-3,17-dione. Compound according to Claim 1, characterized in that it is 1,7a-dimethylandrosta-1,4-diene-3,17-dione or 1,76-dimethylandrosta-1,4-diene-3,17-dione. Compound according to Claim 1, characterized in that it is 1,6a-dimethylandrost-1,4-diene-3,17-dione. 5. A compound according to claim 1, which is 1-ethyl-androsta-1,4-diene-30,17-dione. Compound according to Claim 1, characterized in that it is 1-ethylandrosta-1,4-diene-3,17-dione. 6. A compound according to claim 1, which comprises 1-acetoxymethyl-androsta-1,4-diene-3,17-dione. 20 84487 Compound according to Claim 1, characterized in that it is 1-acetoxymethylandrost-1,4-diene-3,17-dione. 7. A compound according to claim 1, which is 1-hydroxymethyl-androsta-1,4-diene-3,17-dione. A compound according to claim 1, characterized in that it is 1-hydroxymethyl-androsta-1,4-diene-3,17-dione. 84487 BC 8. A compound according to claim 1, which comprises 1-ethoxymethyl-androsta-1,4-diene-3,17-dione. Compound according to Claim 1, characterized in that it is 1-ethoxymethylandrost-1,4-diene-3,17-dione. Composition for use in contraception, characterized in that it contains a compound according to one of Claims 1 to 8. i9 84487 1. 1-alkyl-androsta-1,4-diene-3,17-dione having an anti-inflammatory effect and having all of the formulas I 5 0 10 * 6 color Rj är en methyl, ethyl-, hydroxymethyl-, C1-3-alkoxy ± methyl or C1-4-alkanoyloxymethyl, 15 R6 is a hydrogen atom or a methyl group and R7 is a hydrogen atom or a methyl group 1 7α- or 7B-ställning. 9. A composition comprising a counter-reception, a transverse approach, an embodiment having a face to the face of claims 1-8.