DD237170A1 - METHOD FOR PRODUCING HEART-ACTIVE 14 BETA-HYDROXY ANDROSTAN GLYCOSIDES - Google Patents

Abstract

The invention aims to develop a process for the preparation of cardiac active 14b-hydroxy-androstane glycosides of general formula I. The process according to the invention is characterized in that androstane derivatives are reacted with an 1-haloacyl sugar in an inert solvent in the presence of a suitable heavy metal salt. The reaction takes place between 0C and the boiling point of the solvent, preferably at room temperature. Subsequently, the acylated glycosides are saponified.

Description

For this 1 page formulas

Field of application of the invention

The invention relates to a process for the preparation of cardiopulmonary-14/3-hydroxy-androstane glycosides of the general formula I (see Appendix), in which R ^{1 is} an optionally substituted monosaccharide radical and R ^{2 is} a hydrogen atom, a hydroxyl or an acyloxy group in a or 0 position. Field of application of the invention is the pharmaceutical industry.

Characteristic of the known technical solutions

Compounds of the general formula I are hitherto unknown. They differ from the known cardioprotective glycosides of the cardenolide or bufadienolide type by the absence of the 17/3 permanent lactone ring, which is replaced by a hydrogen atom, a hydroxyl or an acyloxy group. Glycosides of cardenolides have already been prepared by reacting cardenolide genols with peracylated 1-bromo sugars in the presence of fetizone reagent, a celite-dispersed silver carbonate (V.Balogh, M.Fetizon and M.Golfier, J.org. Chemistry 36 [1971] , 1339-1341), and subsequent saponification of the acylated glycosides, for example by treatment with ammonia / methanol or aqueous methanolic potassium bicarbonate solution (H.P. Albrecht, Liebigs Ann. Chem. 1977, 1429-1434). For example, digitoxigenin-aL-rhamnopyranoside (Brown, J. Boutagy and R. Thomas, Arzneimittel. Forsch. [Drug Research] 31 [II], 1059-1064 [1981]) and digitoxigenin-aL -arabinofuranoside (K. Schwab and B. Tschiersch, Pharmazie 37 [1982], 827-828). The production of glycosides of Androstanreihe by this method is new. The starting compounds of general formula II (see Appendix) are not yet known. Only the reaction steps in the synthesis of the compounds of general formula II from compounds of general formula III (see Appendix) are known from other methods.

Object of the invention

The object of the invention is to find a preparation process for compounds of the general formula I and thus to open up a new approach to the development of medicaments with improved application properties in the treatment of cardiac insufficiency.

Explanation of the essence of the invention

According to the invention, compounds of the general formula I are obtained by reacting compounds of the general formula II in which R is a hydrogen atom or an acyloxy group in the a- or / 3-position, with an O-acylated 1-halo-sugar, preferably tribenzoyl-aL-rhamnosylbromide, in the presence of heavy metal salts in an inert solvent.

Inert solvents are e.g. Benzene, toluene, xylene, dioxane, 1,2-dichloroethane or mixtures thereof. As heavy metal salts, especially silver salts such as silver carbonate or silicate are used, which are optionally on a support, preferably diatomaceous earth, finely divided, z. B. Fetizone reagent.

The reaction is carried out at temperatures between 0 ^° C. and the boiling point of the solvent, preferably at room temperature.

Preferably, two to five moles of 1-haloacyl sugar and four to sixteen moles of heavy metal salt are used per mole of steroid, with both the 1-haloacyl sugar and the heavy metal salt being added in portions.

The saponification of the acylated glycosides is preferably carried out by adding 10% aqueous potassium bicarbonate solution in small portions at temperatures between 2O ⁰ C and the boiling point of the solvent, preferably in the range between 40 and 65 ⁰ C, in methanolic solution. The hitherto unknown starting steroids of the general formula II are obtained by known methods from known steroids of the general formula III. Thus, the 3 / 3,14,17 ^ -trihydroxy-5 ^, 14j3-androstane-17-acetate is obtained from 3/3-hydroxy-5/3-androst-14-en-17-one by first adding the 3 / 3-OH group is reacted with dihydropyran to tetrahydropyranyl (THP) ether and then the 17-keto group is reduced with sodium borohydride. After treatment with m-chloroperbenzoic acid, the 17/3-OH group of the resulting 14.1-δ-epoxide is acetoated with acetic anhydride / pyridine and the THP group is cleaved off in alcoholic solution with pyridinium p-toluenesulfonate. Similarly, the corresponding 17a isomer of 3 / 3,14-dihydroxy-5 / 3-14β-androstan-17-one is prepared by reduction of the 17-keto group of the 3/3-THP ether, acetylation of the resulting 17a OH group and subsequent ether cleavage. The 3 / 3,14-dihydroxy-5 / 3,14 / 3-androstane is obtained from the same starting steroid by oxidation of the 17-hydrazone with iodine in triethylamine-tetrahydrofuran, reduction of the resulting vinyl iodide with sodium and subsequent hydrogenation of the A ¹⁶ . Double bond with Pd / activated carbon.

It is surprising and unpredictable that, in the synthesis of the compounds of the general formulas I and II, the tertiary OH group at C-14, which is known to be highly prone to elimination in various other glycosidation reactions (see the overview of WWZorbach and KV Bhat, Adv. Carbohydr. Chem. 21 [1966], 273-321), is not or is not attacked to any significant extent.

It is equally surprising that in the described method, the 3/3-OH group of the steroids used is not or only slightly oxidized, although the used Fetizon reagent as excellent oxidizing agent for SteroidalkoholegiltfM.Fetizon and M.Golfier, C.R.Acad. Be., Ser. C267,900 [1968]).

It is furthermore surprising and unpredictable that the compounds of general formula I obtained by the method, assessed according to their inhibitory effect in the Na, K-ATPase test (K. Repke and H. J. Portius, Experientia 19, 452 [1963]), have cardiac activity. So z. B. the 3/3-aL-rhamnopyranoside of 3ß, 14,17 ^ -trihydroxy-5ß, 14ß-androstane-17-acetate on the Na, K-ATPase from human heart muscle cells approximately 50 times higher molecular biology than the aglycone ( the concentrations required for half maximum inhibition are 4.6 and 232μ.Μ respectively).

The indicated Rr values refer to the thin-layer chromatography on TLC aluminum foil Kieselgel 60 F ₂ 54 (Merck) with the eluent chloroform-methanol (8: 2).

The process is described by the following examples without being limited thereby.

example 1

1.1 g of 3/3-hydroxy-5/3-androst-14-en-17-one (1) in 22 ml of dichloromethane are stirred with 2 ml of dihydropyran and 50 mg of pyridinium-p-toluenesulfonate for 2 hours at room temperature. The solution is then diluted with 30 ml of dichloromethane, washed with saturated aqueous sodium bicarbonate solution, dried over magnesium sulfate and concentrated. The oily residue is chromatographed on 30 g of silica gel (Merck, 100-200mesh). Elution Hexane-acetone (92: 8) gives 1.3 g of 3/3-tetrahydropyranyl ether 2 as an oil.

Nuclear Magnetic Resonance Spectrum (CDCl ₃ ) 10 5.44 (1H, m, Wh / 2 5Hz, 15-H), 4.56 (1H, m, Wh / 2 7Hz, THP-H), 3.90 (1H, brs, 3-H), 4.15-3.15 (2H, m, THP-H), 2.90 (2H, brs, 16H), 1.06 and 0.96 ppm (6H, 2s, 18-H ₃ and 19-H _3).

To dissolve 1.3 g of 2 in 80 ml of methanol-water (9: 1), 440 mg of sodium borohydride are added in portions within 2.5 hours. When the reaction is complete (TLC control), 200 ml of chloroform are added, the solution washed with water (3 × 20 ml), dried over MgSO4 and concentrated. The oily residue is taken up in 35 ml of chloroform and treated for 4 hours at 0-14 ⁰ C with 1.24 g of m-chloroperbenzoic acid. The mixture is then diluted with 65 ml of chloroform, washed with 5% aqueous sodium hydroxide solution (4 × 20 ml) and water (2 × 30 ml). After drying over MgSO ₄ , the solution is concentrated under reduced pressure and the residue is chromatographed on 30 g of silica gel (Merck, 100-200mesh). Elution with hexane-acetone (94: 6) gives (in addition to 270 mg of the corresponding 14a, 15a epoxide) 738 mg of 3 / 3,17 / 3-dihydroxy-14,15 / 8-epoxy-5 / 3,14 / 3. androstane-3-tetrahydropyranyl ether (3).

IR spectrum (Nujol): 3480cm " ¹ (OH).

NMR spectrum (CDCl ₃ ): δ 4.57 (1H, m, Wh / 2 7 Hz, THP-H), 3.90 (1H, br s, 3-H), 4, 10-3, 10 (3H, m, THP-H and 17a-H), 3.50 (1H, s 15-H), 1.07 and 0,97ppm (6H, 2s, _18-H3 and _19-H3 ). 591 mg of 3 in 15 ml of tetrahydrofuran are treated at room temperature with 531 mg of lithium aluminum hydride. After 2 hours, the excess LiAlH _{4 is} reacted with methanol and the mixture, after addition of water, is extracted with toluene. The organic phase is washed with water (3 × 30 ml), dried over MgSO ₄ and concentrated. The residue (570 mg) is acetylated with 6 ml of acetic anhydride in 2.5 ml of pyridine and, after usual work-up, gives 596 mg of oily S & M -dihydroxy-N-acetoxy-O & M / S-androstane-S-tetrahydropyranyl ether (4).

NMR spectrum (CDCl ₃ ): 84.98 (1H, d, J7Hz, 17a-H), 4.72 (1H, m, Wh / 2 8Hz, THP-H), 4.05 (1H, brs, 3H), 4.15 to 3.30 (2H, m, THP-H), 2.12 (3H, s, CH ₃ CO), 1,02ppm (6H, s, _18-H3 and 19- H ₃ ).

300 mg of 40 mg of Pyidin-p-toluenesulfonate in 20 ml of ethanol are added. The solution is refluxed for 30 min.

After cooling, 50 ml of chloroform are added, the solution washed with cold water (2 × 50 ml _), dried over MgSO ₄ and concentrated. Crystallization of the crude product (200 mg) from acetone-hexane gives 140 mg of 3 / 3,14,17 / 3-trihydroxy-5 / 3,14 / 3-androstane-17-acetate (5).

Q .: 138-139 ⁰ C.

IR spectrum (CHCl ₃ ): 3600 (OH), 1740 and 1 240 cm ^-1 (acetoxy).

NMR spectrum (CDCl ₃ ): δ 4.97 (1H, d, J 7Hz, Ma-H), 4.19 (1H, m, Wh / 2 8Hz, 3-H), 1.03ppm (6H , s, 18-H ₃ and 19-H ₃ ). C ₂₁ H ₃₄ O ₄ Ber. C71.96 H 9.78

Gef. C 71.89 H 10.06

100mg (0.28mmol) of 5 in 10ml anhydrous benzene are mixed with 660mg of Fetizon reagent (equivalent to 1.12mmol of Siibercarbonate) and 150mg (0.28mmol) of 2,3,4-tri-O-benzoyl-aL-rhamnopyranosylbromide in 2ml of benzene mixed and stirred under exclusion of atmospheric moisture and light at room temperature. At intervals of 30 min., 660 mg Fetizone reagent and 150 mg Tribenzoyl rhamnosylbromide are added three times. After a total reaction time of three hours, the mixture is filtered and the filtrate is evaporated to dryness in vacuo. The residue is dissolved in 10 ml of methanol and heated to 50-60 ⁰ C in a water bath. With stirring, a total of five times 0.2 ml of 10% strength aqueous potassium hydrogen carbonate solution are added at intervals of 30 minutes. When the reaction is complete after about 3 hours (TLC check), the solution is largely concentrated in vacuo, the residue dissolved in 20 ml of ethyl acetate and washed neutral with water. The organic phase is dried over Na ₂ SO ₄ and chromatographed on 15 g of silica gel 60 (Merck, 70-230 mesh). Elution with chloroform-ethyl acetate (1: 1) gives 62 mg of syrupy crude glycoside, which from dichloromethane-hexane 44mg (31% of theory) crystalline 3β-O- (αL-rhamnopyranosyl) -14,17 / 3-dihydroxy-5i3, 14/3 androstane 17-acetate (6). Q .: 212-215 ⁰ C.

IR spectrum (KBr): 3460.1720.1445.1 375.1 250.1 045crrT ¹ . NMR spectrum (d _e -DMSO): δ 1.97 (3H, s, CH ₃ CO), 1.10 (3H, d, J 6Hz, 6'-H ₃ ), 0.87 and 0.83 ppm (6H, 2s, 18-H ₃ and 19-H ₃ ). ES mass spectrum (high resolution): inter alia 478.2923 (M ⁺ -H ₂ O), 436.2821 (M ⁺ -acetic acid), 332.2359 (genin-H ₂ O), 147.0657 m / z (C ₆ H ₁₁ O ₄ )

 DC: R, = 0.46.

By further elution of the column with chloroform-ethyl acetate (1: 2 to 1: 4), a mixed fraction (88 mg) is obtained, which in addition to the 17/3-acetoxy derivative 6 the corresponding. 17/3-OH compound 7 im Ratio 1: 1 contains. The mixture is treated in 20 ml of methanol under reflux in the manner described four times with 1 ml of 10% potassium bicarbonate solution and gives after usual workup and crystallization from dichloromethane-methanol-hexane 65mg (50% of theory) 3/3 -O- (α-rhamnopyranosyl) -14,17 / 3-dihydroxy-5 / 3,14 / 3-androstane (7)

 F.:226-230°C.

IR spectrum (KBr): 3410.1445.1 375.1 045crrT ¹ .

NMR spectrum Id ₆ -DMSO): δ 1.10 (3H, d, J 6Hz, 6'-H ₃ ), 0.93 and 0.89 ppm (6H, 2s, 18-H ₃ and 19-H ₃ ) , ES mass spectrum (high resolution): inter alia 463.2826 (M ⁺ -H ₂ O), 290.2253 (Genin -H ₂ O), 147.0660 m / z (C ₆ H ₁₁ O ₄ ). DC: Rf = 0.38.

Example 2

To a solution of 380 mg of 3 / 3,14-dihydroxy-5 / 3,14 / 3-androstan-17-one (8) in 30 ml of dichloromethane is added successively 2 ml of dihydropyran and 40 mg of pyridinium p-toluenesulfonate. The reaction mixture is stirred for 4 hours at room temperature, then washed with saturated aqueous sodium bicarbonate solution (2 x 20 ml), dried over MgSO ₄ and concentrated to dryness in vacuo. The residue is chromatographed on 20 g of silica gel (Merck, 100-200mesh). Elution with hexane-acetone (9: 1) ergibt315mg 3 S _j, 14-dihydroxy-5 / 3.14 _/ 8-androstan-17-one-3-tetrahydropyranyl ether (9).

NMR spectrum (CDCl ₃ ): 64.60 (1H, brs, Wh / 2 8Hz, THP-H), 3.93 (1H, brs, 3-H), 4,10-3,10 (2H , m, THP-H), 2.32 (2H, m, Wh / 2 6Hz, 16-H), 1.02 and 0.95ppm (6H, 2s, 18-H ₃ and 19-H ₃ ) 315mg 9 are dissolved in 20 ml of methanol and treated with 184 mg of sodium borohydride. After 2.5 hours, water is added and allowed to crystallize. The precipitate is washed with water, dried and acetylated with 2 ml of acetic anhydride in 1 ml of pyridine for 13 hours. The usual work-up gives 300 mg of oily crude acetate. 10. NMR spectrum (CDCl ₃ ): δ 5.12 (1H, t, J 7 Hz, 17/3-H), 4.60 (1H, m, Wh / 2 8Hz, THP-H), 3.90 (1 H, br s, 3H), 4.15 to 3.20 (2H, m, THP-H), 2.00 (3H, s, COCH ₃₎ , 0.97 and 0,93ppm (6H, 2s, 18-H ₃ and 19H _3).

300mg 10 are mixed with 23 mg pyridinium p-toluenesulfonate in 20 ml of ethanol, the resulting solution heated for 2 hours at 50-60 ° C and then worked up analogously to Example 1. The crude product (270 mg) is recrystallized twice from acetone-hexane to give 160 mg of 3 / 8,14,17a-trihydroxy-5-y-3,14-y-3-androstane-17-acetate (11). F .: 159-161 ⁰ C.

IR spectrum (CHCl ₃ ): 3600 and 3440 (OH), 1725 and 1 240 cm ^-1 (acetoxy).

NMR spectrum (CDCl ₃ ): δ 5.23 (1H, t, J 7Hz, 17/3-H), 4.20 (1H, m, Wh / 2 8Hz, 3-H), 2.10 (3H, s, COCH _3), 1.03 and 1,00ppm (6H, 2s, 18-H ₃ and 19-H _3).

C ₂ iH ₃₄ O ₄ Ber. C71.96 H 9.78

Gef. C 72.07 H 10.02

100 mg 11 are analogously to Example 1 zu53mg (37% d.Th.) 3 / 3-0- (o, -L-rhamnopyranosyl) -14,17a-dihydroxy-5 ^, 14/3-androstan-17-acetate (12 R _f = 0.44) and 45mg (35% of theory) of 3/3-O- (aL-rhamnopyranosyl) -14,17a-dihydroxy-5β, 14/3 androstane (13; R _f = 0 , 36) implemented.

Example 3

To a solution of 800 mg of 8 in 23 ml of 95% ethanol, 11 ml of triethylamine and 13 ml of 80% hydrazine are added and the mixture is refluxed for 3 hours. It is then diluted with water and allowed to crystallize. The crystalline crude 17-hydrazone is dried in air and suspended in a mixture of 15 ml of tetrahydrofuran and 2 ml of triethylamine. With slow stirring, iodine is added in small portions until the solution turns brown. The reaction mixture is diluted with 100 ml of chloroform, with aqueous sodium thiosulfate solution (30 ml) and water (2 x 30 ml).

washed and dried over MgSC> 4. After concentration of the solution on a rotary evaporator remains a crystalline residue (911 mg), which is recrystallized twice from acetone.

Yield: 800 mg 3 / 3,14-dihydroxy-17-iodo-5 / 3,14 / 3-androst-16-ene (14).

F .: 188-189 ⁰ C.

IR spectrum (Nujol): 3450 (OH), 1 600 cm ^-1 (C = C).

Nuclear Magnetic Resonance Spectrum (CDCl ₃ ): δ 6.18 (1H, m, Wh / 2 6Hz, 16-H), 4.18 (1H, m, Wh / 2 7Hz, 3a-H), 1.00 and 0.93ppm (6H, 2s, 18H ₃ and

ES mass spectrum: 416 (M ⁺ ), 289 (M ⁺ -J), 271 (289-H ₂ O), 253 m / z (289 -2H ₂ O).

C ₁₉ H ₂₉ O ₂ J Ber. C 54.82 H 7.01

Gef. C 54.85 H 7.23

To a boiling solution of 911 mg of 14 in 70 ml of anhydrous ethanol, 8 g of sodium are added portionwise over 3 hours. The reaction mixture is diluted with 30 ml of ethanol, neutralized after cooling with 2 N hydrochloric acid and then extracted with 100 ml of chloroform. The organic phase is washed with water (3 × 30 ml), dried over MgSO ₄ and concentrated. The residue gives 600 mg of 3 / 3,14-dihydroxy-5 / 3,14 / 3-androst-16-ene (15).

130 mg of 15 in 12 ml of ethyl acetate are dissolved in a hydrogen atmosphere in the presence of 40 mg 5%. Pd / activated carbon stirred vigorously until after about 4 hours no more hydrogen is absorbed. The catalyst is filtered off and the filtrate is concentrated to dryness. The residue (130 mg) is recrystallized three times from acetone-hexane to give 100 mg of 3 / 3,14-dihydroxy-5 (8,14 / 3-androstane (16).

Q .: 155-156 ⁰ C.

IR spectrum (CHCl ₃ ): 3600 (tert-OH), 3460 cm ^-1 (sec-OH) NMR spectrum (CDCl ₃ ): 54.20 (1 H, m, Wh / 2 8 Hz, 3-H) , 1,00ppm (6H, s, _18-H3 and _19-H3).

ES mass spectrum: 292 (M ⁺ ), 274 (M ⁺ -H ₂ O), 259 (274 -CH ₃ ), 250m / z (M ⁺ -42). C ₁₉ H ₃₂ O ₂ Ber. C 78.03 H 11.03

Gef. C78,10 H 11,28

82 mg (0.28 mmol) of 16 are reacted analogously to Example 1. The reaction mixture is worked up in the manner described and after column chromatography on silica gel 60, in addition to 8 mg of 16, initially 16 mg (15% of theory based on reacted 16) of 3 / 3-0- (aL-rhamnopyranosyl) -5 / 3 are obtained. androst-14-en (17)

Q .: 170-174 ⁰ C.

IR spectrum (KBr): 3405, 3045, 1440, 370, 045 cm " ¹ .

NMR spectrum (CDCl ₃ ): δ 5.10 (1H, d, J 1.6Hz, 15-H), 1.28 (3H, d, J 6Hz, 6'-H ₃ ), 0.98 and 0,93ppm (6H, 2s, 18-H ₃ and 19-H _3). ES mass spectrum (high resolution): 420.2865 (M ⁺ ), 257.2264 (Genin), 147.0663m / z (C ₆ H ₁₁ O ₄ ). DC: R _f = 0.48.

Further elution gives 47 mg (42% of theory based on 16 reacted) of amorphous 3β-O- (α; -L-rhamnopyranosyl) -14-hydroxy-5 / 3-14 / 3-androstane (18)

F .: 136-138 ⁰ C.

IR spectrum (KBr): 3415, 1445, 1375, 1045 cm " ¹ .

NMR spectrum (CDCl ₃ ): δ 1.27 (3H, d, J 6Hz, 6'-H ₃ ), 0.95 and 0.91 ppm (6H, 2s, 18-H ₃ and 19-H ₃ ) ,

ES mass spectrum (high resolution): 438.2985 (M ⁺ ), 420.2874 (M ⁺ - H ₂ O), 275.2368 (Genin), 147.0661 m / z (C ₆ H ₁₁ O ₄ ). DC: R _f = 0.41.

Claims (6)

Invention claim: 1. A process for the preparation of compounds of general formula I, characterized in that an Androstanderivat of the general formula II is reacted in an inert solvent in the presence of a suitable heavy metal salt with a 1-haloacyl sugar, the acyl radicals are then saponified. 2. The method according to item 1, characterized in that serve as the solvent benzene, toluene, xylene, dioxane, 1,2-dichloroethane or mixtures thereof. 3. The method according to item 1 and 2, characterized in that serve as heavy metal salts silver salts such as the carbonate or silicate, which are optionally on a support, preferably diatomaceous earth, finely divided. 4. The method according to item 1 to 3, characterized in that as 1-haloacylated sugar z. B. Tribenzoyl-a-L-rahmnosylbromid is used. 5. The method according to item 1 to 4, characterized in that the reaction between 0 ° C and the boiling temperature of the solvent, preferably at room temperature, is performed. 6. The method according to item 1 to 5, characterized in that the saponification of the acylated glycosides in methanolic solution by adding portions of 10% aqueous potassium bicarbonate solution at temperatures between 20 ⁰ C and the boiling temperature of the solvent, preferably in the range between 40 and 65 ⁰ C, takes place.