Title of Invention

"PROCESS FOR PREPARING HALOGEN-SUBSTITUTED AND PSEUDOHALOGEN SUBSTITUTED 17-METHYLENE-4-AZASTEROIDS"

Abstract

Process for preparing Halogen-substituted and pseudohalogen-substituted 17-methylen-4-azasteroids of general formula I whereinR20 and R20a seperately denote fluoro-, chloro-, bromo-, azido-, rhodano-, Cl-C4-alkyl or hydroxy-Cl-C4-alkyl, R10 denotes a hydrogen atom or methyl group, R4 is selected from the group consisting of hydrogen and a Cl-C4-alkyl group and R1 and R2 together denote a hydrogen atom or an additional bond, characterized in that a 17-methylensteroide of general formula II R20 denote fluoro-, chloro-, bromo-, azido-, rhodano-, C1-C4-alkyl or hydroxy-Cl-C4-alkyl, R10 denotes a hydrogen atom or methyl group is made to react in a known manner with NalO4 under catalytic mediation of KMnO4 in a protic solvent to form the 3,5-secoketo acid, said acid is cyclized with NH4OAc in glacial acetic acid to form the unsaturated lactam and said lactam is then reduced with HCOOH in Dimethyl formamid to the saturated lactam of general formula III wherein the substituents R20 and R10 are the same as in formula II, furthermore reacting the compounds of general formula III with Mel and NaH to form the 4-methyl-substituted compounds of general formula IV wherein the substituents R20 and R10 are the same as in formula II.

Full Text

(Figure Remove) wherein R20 and R30* separately denote fluoro, chloro, bromo and cyano groups, R and R denote a hydrogen atom or a methyl group, and R1 and R2 together denote a hydrogen atom or an additional bond. Methods for preparing novel compounds and for producing pharmaceutical compositions thereof are described. The compounds of the invention, 17-methylene~4-azasteroids, are new, their preparation and their biological activity have thus far not been described. Compounds preferred according to the invention are indicated in Claim 3. Another object of the present invention are pharmaceutical compositions containing as the active ingredient at least one 17-metnyIene-4-azasteroid of general formula I, said compositions optionally containing appropriate auxiliary agents and carriers. The compounds of the invention are 5a-reductase inhibitors. Hence, they are suitable for the treatment of diseases caused by elevated testosterone levels and finally dihydrotestosterone levels in the blood and tissues. Diseases induced by excessive androgen effects can also occur at normal blood testosterone levels if the conversion of testosterone to dihydrotestosterone in the tissues is elevated. This Is the case, for example, in idiopathic forms of hirsutism (REF). Progesterone plays an important role in the tight closing of the os of uterus (Mahendroo). The softening thereof before giving birth is a result of local 5ct-reductase-indiic«d degradation of progesterone to dflrydroprogesterone, which is a very weak gestagen. By inhibiting the progesterone cataboUsm in this part of the uterus, the substances of the invention are therefore also suitable for preventing premature maturation and opening of the os of uterus. 5o>Reduced metabolites of progesterone (REF from Lancet) and other C21-steroids and the metabolites thereof formed in me body, for example aUo-pregnaBolone, can act as neurosteroids and can interact with neurosteroids. Disorders of this function can result in depression. Possible indications for the substances of the invention are prostate diseases, alopecia of the masculine type, acne and hirsutism as well as various gynecological clinical conditions such as the premenstrual syndrome. The appearance of Sa-reduced metabolites of progesterone in the CNS plays an important role in the onset of such conditions. Premature opening of the os of uterus can be induced by increased degradation of progesterone to dihydroprogesterone in this tissue. The substances of the invention are suitable for preventing this catabolism and thus the premature maturation of the cervix uteri. The substances of the invention can exert their action by inhibiting the Sa-reduction of testosterone or progesterone in the organs and tissues affected by this disorder. In addition, the blood levels of the So-reduced metabolites are lowered. Moreover, the compounds of the invention constitute intermediates for the synthesis of other pharmacologically highly active steroid products. The compounds of the invention are prepared as indicated in Claims 4 and 5. According to the invention, the compounds of Claim 1 can be derived from the 17-methylenesteroids of general formula II and general formula VH- (Figure Remove) furthetmore reacting the compounds of general foimula ffl with Mel and NaH in a dipolar aprotic solvent, preferably DMF, to fonn the 4-methyl-substituted compounds of general formula FVand reacting other compounds of general formula DI by a silyl-mediated DDQ (2,3-dichloro-5,6-dicyano-l,4-beo2oquinone) oxidation in a dipolar aprotic solvent such as dioxane to form the dehy-drogenated lactams of general formula V (Figure Remove) The compounds of general formula V, like the compounds of general formula HI, arc reacted with Mel and NaH to form the 4-methyl-substituted lactams of general formula VI (Figure Remove) Moreover, compounds according to Claim 1 are obtained by subjecting compounds of general formula VII to the same chemistry as the compounds of general formula II thus forming compounds of general formulas Vm, DC, X and XI. (Figure Remove) Evaluation of antiandrogenic activity in castrated male rats (see figures/table). Castrated immature male rats were treated for 7 days with testosterone p«»pionale (TP) and the substances of the invention (0.1 mg of TP alone and/or 1 mg of 5a-reductase inhibhor/anlrnal/day, s.c., n = 5 -10/group). The treatment with TP caused a severe increase in weight of the accessory genital glands (prostate and seminal vesicle). In the animals treated with the vehicle and with 5a-reductase inhibitor alone, the weights of the examined organs remained at low values. We found that the substances of the invention exerted an inhibitory effect hi the teat systems selected. Table XX [sic] shows that the compounds J 1879 and J 1924 weakened appreciably the effects of TP on the prostate and seminal vesicle. Table: Inhibition of the action of testosterone propionate (TP) on the growth of the seminal vesicle and prostate. Test on sexually immature, castrated male rats, n = 5 - 10/group, dosage: TP: 0.1 mg/animal/day s.c., Sa-reductase inhibitor, 1 mg/animal/day s.c., difference vehicle controls and. TP controls * 100%. (Figure Remove) The studies to determine the ICso values of the steroids J 1879 and J 1924 for the 5a-reductase outside the genital tract were carried out in four different human bone cell lines (hOB cells). The cells were incubated for 6 hours with 0.5 uM androstendione (0.1 joM [*H] androstendionc; 0.4 nM androstendione) with or without adding increasing concentrations of the inhibitor (10"11 - \ 0 M). After the incubation, the medium was extracted with chloroform : metbanol (2 : 1, vol: vol), the steroids (substrate and So-reduced metabolites) were separated by thin-layer chromatography, and the DNA content of the cells of the samples was determined. The 5a-reductase activity (sum of the So-reduced metabolites formed: 5a-androstandione, Sa-dihydrotestosterone, 3a-androstan-3a,17p-diol, 5a-androstan-3p,17p-diol expressed inpmol/jig DNA/h) was determined in duplicate samples. The relative So-reductase activity (Sa-reductase activity of the samples with added inhibitor compared to the corresponding control values, namely those for the samples without added inhibitor) is shown in the figure always as a mean ± SEM. The ICso values of J 1879 and J 1924 for the Sa-reductase in bone cells amounted to In addition, within the framework of studies to determine the ICso values of J 1879 and J 1924, the inhibiting action of LY 191704, a nonsteroidal specific inhibitor of type-1 5a-reductase. finasteride, a 4-azasteroid with inhibiting action mainly on type-2 5a-reductase, and progesterone, a physiological substrate of 5a-reductase, was determined for comparative purposes in all four cell toes at a concentration of 10^ M. It can be seen from the figures that the substances of the invention inhibited the fa-reductase in the cell types studied more strongly and at substantially lower concentrations than did the said natural substrates and reference substances. The invention will be illustrated in greater detail by the following examples. EXAMPLE 1 E-l 7-ChloromethyleMe-4-HMi-5a-aDdrostan-3-one To a solution of 3.1 mmol (1,0 g) of E-17-chloromethylene-4-aza-androst-5-cn-3-one in 140 mL of dimethylformanude were added with stirring 59 mL of formic acid (85%) and 115.8 mmol (16 g) of potassium carbonate, and the reaction mixture was heated at reflux for 8 hours. Toluene was then added to the reaction solution, and ihe mixture was evaporated under vacuum. The residue was taken up in water, and the resulting mixture was extracted with methylene chloride. The combined extracts were treated with saturated sodium carbonate solution, washed neurral with water, dried over sodium sulfste and evaporated. The resulting crude product was crystallized from acetone/n-hexane, which gave 543 mg of solid product (54%). M.p. = 148 - 151 °C; fa]D2° = +16° (CHCb). EXAMPLE! E-17-Chloromethyleac^4-methyl-4-aza-5o-androstan-3-one To a suspension of 1.06 mmol (340 mg) of B-17^Uoromethylene^-^za-5a-androstan-3-one in 9 mL of dimethylforraamide were added at room temperature and under an argon atmosphere 3.08 mmol (123 mg) of sodium hydride (60% in oil). The reaction mixture was stirred 30 min, and to it was then added dropwise a solution of 5.3 mmol (0.33 mL) of methyl iodide in 3.0 mL of dimethylformamide. After about 60 minutes, 2 mL of methanol was added followed after another 10 min by 9 mL of saturated aqueous ammonium chloride solution. The reaction mixture was diluted with water and extracted with toluene. The combined extracts were washed with water, dried over sodium sulfate and evaporated- The resulting crude product was crystallized from acetone/n-hexane, which gave 154 mg of solid product (43%). M.p. -150 - 162 °C; [a]D20 = -7- (CHCb). EXAMPLES E-17-CUoromethylette~4~aza-5a>aadrost-l-«n-3-«ne 1.34 mmol (430 mg) of E-17-chloromethylene-4-aza-5a-androstan-3-one was suspended in 9 mL of dioxane at room temperature and under an argon atmosphere and to the suspension were then added 1.5 mmol (340 mg) of 2.3^ichloro-5,6^cyano-p-benzoquinone and 6.4 mraol (1.7 mL) of bis- (trimethy!silyl)trifluoroac«ainide. The mixture was steed first 3 hours at room temperature and then 3 hours in an oil bam at about 100 -110 °C. The reaction solution was diluted with methylene chloride and then washed first with 2% aqueous sodium hydrogen sulfite solution, then with 2N hydrochloric acid and finally with water. The remaining extract was dried over sodium sulfate and evaporated. Crystallization from acetone gave 243 mg of solid product (57%). M.p. = 275 - 282 °C; [a]D2° = -41° (CHCfe). EXAMPLE 4 E-17-Chloromethyl«ne-4-metliyl-4-aza-5o-androst- To a suspension of 0.87 mmol (279 mg) of E-17-^oromethylene^-aza-5a-androst-l -en-3-one in 7 mL of dimcihylfbrmamide were added at room temperature and under an argon atmosphere 2.4 mmol (98 mg) of sodium hydride (60% in oil). The reaction mixture was stirred for 30 minutes, after which a solution of 7.5 mmol (0.47 mL) of methyl iodide in 3 mL of dimethylformamide was added dropwise. After about 60 minutes, 2 mL of methanol was added followed after an additional 10 min by 9 mL of saturated aqueous ammonium chloride solution. The reaction mixture was diluted with water and extracted with toluene, and the extract was washed with water, dried over sodium sulfate and evaporated. The resulting crude product was crystallized from ethyl acetate, which gave 122 mg of solid product (42%). M.p. = 160 - 165 °C; [a]o20 = -47° (CHCk). EXAMPLES E-17-ChIoromcthylene-4-a2a-5a-«stran-3-one To a suspension of 327 mmol (1 g) of E-17 EXAMPLE 6 E-17-Chloromethylene^4-methyl-4-aza-5a-wtrfln-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 dimethylfbimainide were added at room temperature and under an argon atmosphere 1.7 mmol (73 mg) of sodium hydride (60% in oil). The reaction mixture was stirred 30 min after which a solution of 3.2 mmol (0.2 mL) of methyl iodide in 2 mL of dimethylfortnamide was added dropwise. After about 60 minutes, 1 mL of methanol was added followed after an additional 10 min by 4 mL of saturated aqueous ammonium chloride solution. The reaction mixture was diluted with water and extracted with methykne chloride. The combined extracts were washed with water, dried over sodium sulfate and evaporated. The resulting crude product was purified by chromaiography on Silica Gel 60 (eluent; methylene chloride/acetone 8/2). Crystallization from ethyl acetate then gave 131 mg of solid product (62%). M.p. = 161-171 °C; [a]b20 = -88° (CHCb). EXAMPLE 7 E-l 7-Chloromethylfcne-4-azA-5a-€Str-l-«j! -3-on e 1 mmol (310 mg) of E-17-chloromethyl«ie-4-a2a-5a-estran-3-one was suspended in 6.3 mL of dioxanc at room temperature and under an argon, atmosphere. Thereafter, 2.3 mmol (522 mg) of 2,3-dichloro-5,6-dicyano-p-benzoquinone and 9.8 mmol (2.6 mL) of Hs^trimethylsilyl)aifluoro-acetamide were added. The mixture was stirred first 3 hours at room temperature and then 15 hours in an oil bath at 100 -110 °C. The reaction solution was diluted with methylene chloride and then washed first with 2% aqueous sodium hydrogen sulfite solution, then with 2N hydrochloric acid and finally with water. The remaining residue was dried over sodium sulfate and evaporated. Purification by chromatography on Silica Gel 60 (eluent: methylene chloride/methancl 98/2) gave 57 mg of solid product (18.5%). [a]D20 = 37« (CHCb). We Claim: 1. Process for preparing Halogen-substituted and pseudohalogen-substituted 17-methylen-4-azasteroids of general formula I (Figure Remove) wherein R20 and R20a seperately denote fluoro-, chloro-, bromo-, azido-, rhodano-, Cl-C4-alkyl or hydroxy-Cl-C4-alkyl, R10 denotes a hydrogen atom or methyl group, R4 is selected from the group consisting of hydrogen and a Cl- C4- alkyl group and R1 and R2 together denote a hydrogen atom or an additional bond. characterized in that a 17-methylensteroide of general formula II (Figure Remove) wherein R20 denote fluoro-, chloro-, bromo-, azido-, rhodano-, Cl-C4-alkyl or hydroxy-C 1 -C4-alkyl, R10 denotes a hydrogen atom or methyl group is made to react in a known manner with NalO4 under catalytic mediation of KMnO4 in a protic solvent to form the 3,5-secoketo acid, said acid is cyclized with NH4OAc in glacial acetic acid to form the unsaturated lactam and said lactam is then reduced with HCOOH in Dimethyl formamid to the saturated lactam of general formula III (Figure Remove) wherein the substituents R20 and R10 are the same as in formula II, furthermore reacting the compounds of general formula III with Mel and NaH to form the 4-methyl-substituted compounds of general formula IV (Figure Remove) wherein the substituents R20 and R10 are the same as in formula II. 2. Process for preparing compounds as claimed in claim 1 wherein R20 denote fluoro-, chloro-, bromo-, azido-, rhodano-, Cl-C4-alkyl or hydroxy-C 1 -C4-alkyl, R20a denotes a hydrogen atom, R10 denotes a hydrogen atom or methyl group, R4 denotes a methylen group, R1 and R2 together denote a hydrogen atom, characterized in that a 17-methylensteroide of general formula II (Figure Remove) wherein R20 denote fluoro-, chloro-, bromo-, azido-, rhodano-, Cl-C4-alkyl or hydroxy-C 1 -C4-alkyl, R10 denotes a hydrogen atom or methyl group and, like the compounds of formula II as claimed in claim 1, reduced to the saturated lactam of general formula III (Figure Remove) wherein the substituents R20 and R10 are the same as in formula II, and by a silyl-mediated oxidation to form the dehydrogenated compounds of general formula V (Figure Remove) wherein the substituents R20 and R10 are the same as in formula II, and reacting the compounds of general formula V with Mel and NaH to form the 4-methyl-substituted compounds of general formula VI (Figure Remove) wherein the substituents R20 and R10 are the same as in formula II. 3. Process for preparing compounds as claimed in claim 1 wherein R20 denotes a hydrogen atom R20a denote fluoro-, chloro-, bromo-, azido-, rhodano-, Cl-C4-alkyl or hydroxy-C 1 -C4-alkyl, R10 denotes a hydrogen atom or methyl group, R4 denotes a methylen group, R1 and R2 together denote a hydrogen atom, characterized in that a 17-methylensteroide of general formula VII wherein (Figure Remove) R20a denote fluoro-, chloro-, bromo-, azido-, rhodano-, C l-C4-alkyl or hydroxy-C 1-C4-a/kYI, R20 denotes a methylene group R10 denotes a hydrogen atom or methyl group and, the unsaturated lactam is reduced to form to the compounds of general formula VIII (Figure Remove) furthermore, reacting the compound VIII with Mel and NaH to form to the compounds of general formula IX (Figure Remove) wherein the substituents R20a and R10 are the same as in formula VII. 4. Process for preparing compounds as claimed in claim 1, wherein R20 denotes a hydrogen atom R20a denote fluoro-, chloro-, bromo-, azido-, rhodano-, Cl-C4-alkyl or hydroxy-C 1 -C4-alkyl, R10 denotes a hydrogen atom or methyl group, R4 denotes a methylen group, R1 and R2 together denote a hydrogen atom, characterized in that a 17-methylensteroide of general formula VII (Figure Remove) wherein R20a denote fluoro-, chloro-, bromo-, azido-, rhodano-, Cl-C4-alkyl or hydroxy-C 1 -C4-alkyl, R20 denotes a methylene group R10 denotes a hydrogen atom or methyl group and, converting into unsaturated lactam to form the compounds of general formula X (Figure Remove) furthermore, dehydrogenating to form the compounds of general formula XI (Figure Remove) wherein the substituents R20a and R10 are the same as in formula VII.

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