MXPA04001822A

MXPA04001822A - Methylene-4-azasteroids.

Info

Publication number: MXPA04001822A
Application number: MXPA04001822A
Authority: MX
Inventors: Sch Llkopf Klaus
Original assignee: Schering Ag
Priority date: 2001-08-28
Filing date: 2002-08-28
Publication date: 2004-06-29

Abstract

Novel 17-methylene-4-azasteroids of general formula (I), where R20 and R20a independently = fluoro, chloro, bromo and cyano groups; R4 and R10 = H or methyl; R1 and R2 = each an H atom or together an additional bond, are disclosed. Said compounds are inhibitors of 5-alpha-reductase and are thus suitable for the treatment of diseases which are the result of increased testosterone and hence dihydrotestosterone levels in blood and tissue.

Description

METILEN-4-AZA-EAST OIDES DESCRIPTION The invention relates to novel 17-methylene-4-aza-steroids of the general formula I where R20 and R20a separately represent fluorine, chlorine, bromine and cyano groups. R4 and R10 denote respectively a hydrogen atom or a methyl group. R1 and R2 are respectively a hydrogen atom or an additional bond. New methods are described for the preparation of new compounds, as well as for the preparation of their pharmaceutical composition. The inventive compounds -17-methylene-4-aza-steroids are new, their preparation and biological activity have not been described so far. Inventively preferred compounds are related in claim 3. Subject of the present invention are furthermore pharmaceutical compounds containing as active substance at least one 17-methylene-4-aza-steroid of the general formula I, optionally containing auxiliary substances and suitable vehicles . The inventive compounds are inhibitors of 5 - . 5 -reductasa. They are suitable, therefore, for the treatment of diseases resulting from increased testosterone and ultimately the level of dihydrotestosterone in the blood and tissue. Diseases resulting from excessive androgenic effects can also occur with normal levels of testosterone in the blood, when the transformation of testosterone to dihydrotestosterone is increased in the tissue. This is the case, for example, in the idiopathic forms of hirsutism (REF). Progesterone plays an important role in the firm closure of the neck orifice (Mahendroo). Its softening before birth is the result of a local reduction of progesterone by 5cx-reductase to dihydroprogesterone, which is a very weak progestin. Through the inhibition of progesterone catabolism in this part of the uterus, the inventive substances, therefore, are also convenient to prevent premature maturation and opening of the neck orifice.

Progesterone metabolites reduced by 5a-reductase (Lancet's REF) and other C21 steroids and their metabolites by developing in the body,. for example, alo-pregnanolone, can act as neuro steroids and interact with neuro-steroids. Alterations of this function may result in impairment of welfare. Possible indications for the inventive substances are diseases of the prostate, alopecia of the masculine type and acne and hirsutism, as well as different gynecological pictures, such as the menstrual syndrome. During its gestation, the presence of progesterone metabolites reduced by 5a-reductase in the central nervous system plays an important role. The premature opening of the orifice of the neck can be induced by the accelerated reduction in these tissues to dihydroprogesterone. The inventive substances are convenient to inhibit this catabolism and with this the premature maturation of the cervix of the uterus. The inventive substances can exert their activity by inhibiting the 5a-reduction of testosterone or progesterone in the organs and tissues affected with the dysfunction. To this is added the decrease in the blood level of the metabolites reduced by 5a-reductase.

The inventive compounds also represent intermediates for the synthesis of other pharmacologically very active steroid products. The preparation of the inventive compounds is carried out according to claims 4 and 5. The compounds according to claim 1 are inventively accessible from the 17-methylene steroids of the general formula II and the general formula VII.

Compounds according to claim 1 are obtained by converting the compounds of the general formula II in a manner known per se with NaI04 with catalytic intermediation by ?? 04 · in a protic solvent, preferably t.-BuOH, in acid 3.5 - ketonic acid, then cyclizing with NH4OAc in glacial acetic acid to obtain unsaturated lactam and reducing with HCOOGH / K2C03 in DMF to obtain saturated lactam of general formula III, then transforming the compounds of the general formula III with Mel and NaH in an aprotic bipolar solvent, preferably DMF in 4-methyl substituted compounds of the general formula IV, as well as on the other hand transforming the compounds of the general formula III into an oxidation of DDQ (2,3-dichloro-5,6-dicyano-1,4-benzoquinone) mediated by silyl in an aprotic bipolar solvent, such as dioxane, in the dehydrated lactams of the general formula V.

The compounds of the general formula V are converted analogously to the compounds of the general formula III with Mel and NaH in the 4-methyl substituted lactams of the general formula CI.

In addition, compounds according to claim 1 are obtained by subjecting compounds of the general formula VII to the same chemical processes as the compounds of the general formula II and then obtaining compounds of the general formula VIII, IX, X and XI.

Results Evaluation of anti-androgenic activity in the castrated male rat (see images / table). Castrated immature male rats were treated for 7 days with testosterone proprionate (TP) and the inventive substances (0.1 mg TP alone and / or with 1 mg of 5a-reductase inhibitor per day / animal sc, n = 5-10 / group ). Treatment with TP caused a large increase in the weights of the associated sex glands (prostate and seminal vesicle). In animals treated with only the vehicle and the 5cc-reductase inhibitor, the weights of the organs investigated remain at low values. It has been found that the inventive substances had inhibitory activity in the selected test systems. Table XX shows that compounds J 1879 and J 1924 clearly reduce the effects of TP to prostate and seminal vesicle. Table: Inhibition of testosterone propionate (TP) activity on seminal vesicle and prostate growth. Test in sexually castrated immature male rats, n = 5-10 / group, dosage: TP 0.1 mg / animal / day s.c, 5a-reductase inhibitor: 1 mg / animal / day s.c. Vehicle difference and TP controls = 100%.

ET 01.31 prostate seminal vesicle 34 25 TP + J 1879 39 41 TP + J 1924 The tests to determine the IC50 values for 5a-reductase by steroids J1879 and J1924 outside the genital tract, were performed on 4 different lines of human bone cells (hOB cells). The cells were incubated for 6 hours in respectively 0.5 μ of androstenedione (0.1 μ? [3 H] androstenedione, 0.4 μ? Androstenedione) with or without the addition of the inhibitors (10-11 - 10 8 8 M) in incremental concentrations. After incubation, the medium was extracted with chloroform: methanol (2: 1; vol.vol) and the steroids (substrate and metabolites reduced by 5a-reductase) were separated with thin layer chromatography and the DNA content of the cells was determined. The activity of 5a-reductase (sum of the metabolites formed, reduced with 5a-reductase: 5a-androstanione, 5a-dihydrotestosterone, 5a-androstan-3ct, 17β-diol, 5a-andostran-3p, 17β- ??? 1 expressed in pinol g DNA / h) was determined in respective duplicate assays.The relative 5a-reductase activity (5oc-reductase activity of the samples with addition of the inhibitors compared to the values of the controls, is say, the samples without addition of inhibitors) FICA image value ± SEM average. The IC50 values of J1879 and J1924 for the 5cc-reductase in bone cells are in each case < 10"10 M. In the framework of the investigations to determine the IC50 values of J1879 and J1924, the inhibitory activity of LY191704, a non-spheroidal inhibitor specific to 5 -reductase type 1, finasteride, 4-5, was determined for comparative purposes. aza-steroid with inhibitory activity mainly of 5a-reductase type 2 and progesterone, a physiological substrate of 5a-reductase, in all 4 cell lines with a respective concentration of 10 ~ 8 M. From the image it turns out that the inventive substances inhibit 5cc-reductase in the cell types investigated the most. strongly and in concentration substantially lower than the natural substrates mentioned and the reference substances. The invention is explained below more closely by the following examples. Example 1 E-17-chloromethylene-4-aza-5a-androstan-3-one? a solution of 3.1 mmol (1.0 g) of E.17-chloromethylene-4-aza-androst-5-en-3-one in 140 ml of dimethylformamide was added with 59 ml of formic acid (85%) and 115.8 mmol (16 g). mg) potassium carbonate under stirring and the reaction mixture is heated to reflux for 8 hours. The reaction solution is then mixed with toluene and concentrated in vacuo. The residue is taken up in water and extracted with methylene chloride. The bound extracts are treated with a saturated solution of sodium carbonate, wash neutral with water, dry over sodium sulfate and concentrate. ?? p ?? d ?? t? ??? d? ? ßte ?? d? Is that ??? a e? ? et? a /? -hexane and 543 mg of a solid product (54%) are obtained Melting point = 148 - 151 ° C; [a] D20 = + 16 ° (CHC13) Example 2 E-17-chloromethylene-4-methyl-4-aza-5 -androstan-3-one To a suspension of 1.06 mmol (349 mg) of E-17-chloromethylene 4-aza-5a-androstan-3-one in 9 ml of dimethylformamide are added at room temperature and under an argon atmosphere, 3.08 mmol (123 mg) of sodium hydride (60% in oil). The reaction mixture is stirred for 30 minutes and then a solution of 5.3 mmol (0.33 ml) of methyl iodide in 3.0 ml of dimethylformamide is added dropwise. After about 60 minutes, 2 ml of methanol are added and after another 10 minutes 9 ml of a saturated aqueous solution of ammonium chloride. The reaction mixture is diluted with water and extracted with toluene. The combined extracts are washed with water, dried over sodium sulfate and concentrated. The crude product obtained is recrystallized from acetone / n-hexane and 154 mg of a solid product (43%) are obtained. Melting point = 150 - 162 ° C; [cc] D20 = -7 ° C (CHC13) 'Example 3 E-17-chloromethylene-4-aza-5a-androst-l-eix-3-one 1.34 mmol (430 mg) of E-17-chloromethylene-4 -za-5a-androstan-3-one are suspended in an argon atmosphere at room temperature in 9 ml of dioxane and then 1.5 mmol (340 mg) of 2,3-dichloro-5,6-dicyano-p-benzoquinone are added. , as well as 6.4 mmol (1.7 ml) of bis- (trimethylsilyl) -trifluoroacetamide. It is stirred first for 3 hours at room temperature and then for 3 hours in an oil bath at approximately 100-110 ° C. The reaction mixture is diluted with methylene chloride and washed, one followed by the other, with an aqueous solution of 2% sodium hydrosulfite, 2N hydrochloric acid and water. The remaining extract is dried over sodium sulfate and concentrated. After recrystallization from acetone, 243 mg of a solid product (57%) are obtained. Melting point = 275-282 ° C; [a] D20 = -41 ° (CHC13) Example 4 E-17-chloromyl ilen-4-methylene-4-aza-5a-androst-1-en-3-one To a suspension of 0.87 mmol (297 mg) of E-17-chloromethylene-4-aza-5a-androst-1-en-3-one in 7 ml of dimethylformamide are added at room temperature and under an argon atmosphere, 2.4 mmol (98 mg) of sodium hydride (60% strength). oil). The reaction mixture is stirred for 30 minutes and then a solution of 7.5 mmol (0.47 ml) of methyl iodide in 3 ml of dimethylformamide is added dropwise. After about 60 minutes, 2 ml of methanol are added and after another 10 minutes 9 ml of a saturated aqueous solution of ammonium chloride. After diluting the reaction mixture with water, it is extracted with toluene, the extract is washed with water, dried over sodium sulfate and concentrated. The crude product obtained is recrystallized from acetic acid and 122 mg of a solid product (42%) are obtained. Melting point = 160 - 165 ° C; [a] D20 = -47 ° (CHC13) Example 5 E-17-chloromethylene-4-aza-5a-estran-3-one To a suspension of 3.27 mmol (Ig) of E-17-chloromethylene-4-aza- 3-on-3-one in 150 ml of dimethylformamide are added 73.3 ml of formic acid (85%) and 121.55 mmol (16.8 g) of potassium carbonate under stirring and the reaction mixture is refluxed for 12 hours. The reaction solution is then mixed with toluene and concentrated in vacuo. The residue is absorbed with water and extracted with methylene chloride. The combined extracts are treated with a saturated sodium carbonate solution, washed neutral with water, dried over sodium sulfate and concentrated. The crude product obtained is recrystallized from acetone / n-hexane and 461 mg of a solid product (46%) are obtained. Melting point = (178-200) 262-282 ° C; [a] D20 = -21 ° (CHC13) Example 6 E-17-chloromethylene-4-methyl-4-aza-5a-estran-3-one To a suspension of 0.65 mmol (200 mg) of E-17-chloromethylene 4-aza-5a-estran-3-one in 5.8 ml of dimethylformamide are added, at room temperature and in an argon atmosphere, 1.7 mmol (73 mg) of sodium hydride (60% in oil). The reaction mixture is stirred for 30 minutes and then a solution of 3.2 mmol (0.2 ml) is added dropwise., of methyl iodide in 2 ml of dimethylformamide. After approximately 60 minutes, 1 ml of methanol are added and after another 10 minutes, 4 ml of a saturated aqueous solution of ammonium chloride. The reaction mixture is diluted with water and extracted with methylene chloride. The combined extracts are washed with water and dried over sodium sulfate and concentrated. The crude product obtained is purified by chromatography on silica gel 60 (eluent methylene chloride / acetone = 8/2) and, after recrystallization from ethyl acetate, 131 mg of a solid product (62%) are obtained. Melting point = 161-171 ° C; [] D20 = -88 ° (CHC13) Example 7 E-17-chloro me ilen-4-aza-5a-estr-l-en-3-one 1 mmol (310 mg) of E-17-chloromethylene-4-methyl 4-aza-5a-estran-3-one are suspended at room temperature in 6.3 ml of dioxane and then 2.3 mmol (522 mg) of 2,3-dichloro-5,6-dicyan-p-benzoquinone are added, as well as 9.8 mmol (2.6 ml) of bis- (trimethylsilyl) -trifluoracetamide. It is stirred first at room temperature for 3 hours and then for 15 hours in an oil bath at 100-110 ° C. The reaction solution is diluted with methylene chloride and washed in this order with a solution of 2% sodium hydrosulphite, 2N hydrochloric acid and water. The remaining extract is dried over sodium sulfate and concentrated. After purification by chromatography on silica gel 60 (eluant chloride of

Claims

CLAIMS 1. 17-methylene-4-aza-steroids substituted with halogens and pseudohalogens of the general formula I, where R20 and R20a separately represent fluorine, chloro-, bromo-, azide-, cyano-, isocyano-, rolino-, isothiocyano-, Ci-4-alkyl or hydroxy-Cx- ^ -alkyl, R10 denotes a hydrogen atom or a methyl group, R4 is selected from a group consisting of hydrogen and Ci-4-alkyl, R1 and R2 are respectively a hydrogen atom or an additional bond. 2. 17-methylene-4-aza-steroids, substituted with halogens and pseudohalogens according to claim 1, where denotes: R4 = H, Me R20 / R20a = F ^ clf BXf CN > 3. 17-methylene-4-aza-steroids according to claim 1, namely E-17-chloromethylene-4-aza-5-aza-estran-3-one E-17-chloromethylene-4-aza-5oc -androstan-3-one E-17 chloromethylene-4-methyl-4-aza-5-tetra-3-one E-17 chloromethylene-4-methyl-4-aza-5a-androstan-3-one E-17 chloromethylene -4-aza-5a-estr-l-en-3-ona E-17 chloromethylene-4-aza-5-androst-l-en-3-one E-17 chloromethylene-4-methyl-4-aza-5cc -androst-l-en-3-one E-17-bromomethylene-4-methyl-4-aza-5a-estran-3-one E-17-bromomethylene-4-methyl-4-aza-5a-estran-3 -one E-17-cyanomethylene-4-methyl-4-aza-5-oc-estran-3-one E-17-cyanomethylene-4-methyl-4-aza-5a-estran-3-one Z-17- (1 ') -chloroethylidene-4-methyl-4-aza-5a-androstan-3-one Z-17- (1') -chloroethylidene-4-methyl-4-aza-5a-estran-3-one Z-17- (1 ') -bromethylidene-4-methyl-4-aza-5a-androstan-3-one Z-17- (1') -Bromethylidene-4-methyl-4-aza-5-tetra-3-one. Process for the preparation of compounds according to claim 1, characterized in that 17-methylene steroids of the general formula II are transformed, where R20 denotes a function of fluorine, chlorine, bromine or cyano and R10 is a hydrogen atom or a methyl group, in a manner known per se with NaI04 with catalytic intermediation of KMn04 in a protic solvent, in 3, 5-dry ketoacid, this is cyclized with? ¾ ??? in glacial acetic acid to form unsaturated lactam and reduced with HCOOH in DMF to saturated lactam of the general formula III with the substituents R20 and R10 as in the formula II, on the one hand, the compounds of the general formula III with Mel and Nati are further transformed into the 4-methyl-substituted compounds of the general formula IV, with the substituents R20 and R10 as in the formula II, as well as, on the other hand, the compounds of the general formula III in a silyl-mediated oxidation, in the dehydrated compounds of the general formula V with the substituents R20 and R10 as in formula II, it produces wherein these compounds of the general formula V are converted analogously to the compounds of the general formula III with Mel and NaH in the 4-methyl-substituted compounds of the general formula VI with the substituents R20 and R10 as in formula II. 5. Process for the preparation of 17-methylene-4-aza-steroids according to claims 1, 2, 3 and 4, characterized in that 17-methylene-steroids of the general formula VII are converted, where R20a denotes a function of fluorine, chlorine, bromine or cyano and R10 is a hydrogen atom or a methyl group, as described in claim 4, to obtain the compounds of the general formula VIII, IX, X and XI. 6. A pharmaceutical composition containing at least one 7-methylene-4-aza-steroid according to claim 1, optionally together with pharmaceutically compatible auxiliaries and carriers. 7. Use of a compound according to claim 1, for preparing a medicament for the treatment of the increased level of androgens. 8. Use of a compound according to claim 1, for preparing a medicament for the treatment of acne.