3-NITROGEN OR SULPHUR SUBSTITUTED OESTRA-1,3,5(10),16-TETRAENE AKR1C3 INHIBITORS

Abstract

The invention relates to AKR1C3 inhibitors of formula (I) and to processes for preparation thereof, to the use thereof for treatment and/or prophylaxis of diseases and to the use thereof for production of medicaments for treatment and/or prophylaxis of diseases, especially of bleeding disorders and endometriosis.

Claims

1. Compounds of the general formula (I) ##STR00107## where: A represents a group selected from: ##STR00108## wherein * indicates the point of attachment of said group with the rest of the molecule; X is a group selected from: ##STR00109## wherein * indicates the point of attachment of the group with the rest of the molecule and where the group is optionally substituted, one or more times, independently from each other, with a substituent selected from halogen, CN, OH, RR.sup.2N—, C.sub.1-C.sub.6-alkyl, C.sub.1-C.sub.6-haloalkyl, C.sub.1-C.sub.6-alkoxy, C.sub.1-C.sub.6-haloalkoxy, wherein C.sub.1-C.sub.6-alkyl and C.sub.1-C.sub.6-haloalkyl groups are optionally substituted with OH; R.sup.1 is C.sub.1-C.sub.6-alkyl or C.sub.1-C.sub.6-haloalkyl, and where R.sup.1 is optionally substituted with one or two substituents, independently from each other, selected from OH, CN, RO(CO)—, RR.sup.2N(CO)—, RR.sup.2NSO.sub.2—, C.sub.1-C.sub.6-alkyl-(CO)(NH)SO.sub.2— or 5-tetrazolyl; R.sup.2, R.sup.5 are, independently from each other, hydrogen or C.sub.1-C.sub.6-alkyl, where C.sub.1-C.sub.6-alkyl groups are optionally substituted, one or more times, independently from each other, with halogen, R.sup.3, R.sup.4 are, independently from each other, C.sub.1-C.sub.6-alkyl or C.sub.1-C.sub.6-haloalkyl, and whereby R.sup.3 and R.sup.4 are optionally substituted with one or two substituents, independently from each other, selected from OH, CN, RO(CO)—, RR.sup.2N(CO)—, RR.sup.2NSO.sub.2—, C.sub.1-C.sub.6-alkyl-(CO)(NH)SO.sub.2— or 5-tetrazolyl; R.sup.6, R.sup.7, R.sup.8, R.sup.9 are, independently from each other, C.sub.1-C.sub.6-alkyl or C.sub.1-C.sub.6-haloalkyl, and whereby R.sup.6, R.sup.7, R.sup.8 and R.sup.9 are optionally substituted with one or two substituents, independently from each other, selected from OH, CN, RO(CO)—, RR.sup.2N(CO)—, RR.sup.2NSO.sub.2—, C.sub.1-C.sub.6-alkyl-(CO)(NH)SO.sub.2— or 5-tetrazolyl; R is hydrogen or a C.sub.1-C.sub.6-alkyl group; or the stereoisomers, tautomers, N-oxides, hydrates, solvates or salts thereof, or a mixture consisting of the above.

2. Compounds of the general formula (I) according to claim 1, where A represents a group selected from: ##STR00110## wherein * indicates the point of attachment of said group with the rest of the molecule; X is a group selected from: ##STR00111## wherein * indicates the point of attachment of the group with the rest of the molecule and where the group is optionally substituted, one or more times, independently from each other, with a substituent selected from halogen, C.sub.1-C.sub.4-alkyl, C.sub.1-C.sub.4-haloalkyl, C.sub.1-C.sub.4-alkoxy, wherein C.sub.1-C.sub.4-alkyl and C.sub.1-C.sub.4-haloalkyl groups are optionally substituted with OH; R.sup.1 is C.sub.1-C.sub.4-alkyl, which is optionally substituted with one or two substituents, independently from each other, selected from OH, RO(CO)—, RR.sup.2N(CO)—, RR.sup.2NSO.sub.2— or 5-tetrazolyl; R.sup.2, R.sup.5 are, independently from each other, hydrogen or C.sub.1-C.sub.4-alkyl, where C.sub.1-C.sub.4-alkyl groups are optionally substituted, one or more times, independently from each other, with halogen; R.sup.3, R.sup.4 are, independently from each other, C.sub.1-C.sub.4-alkyl, and whereby R.sup.3 and R.sup.4 are optionally substituted with one or two substituents, independently from each other, selected from OH, RO(CO)—, RR.sup.2N(CO)—, RR.sup.2NSO.sub.2—, C.sub.1-C.sub.4-alkyl-(CO)(NH)SO.sub.2— or 5-tetrazolyl; R.sup.6, R.sup.7, R.sup.8, R.sup.9 are, independently from each other, C.sub.1-C.sub.4-alkyl, and whereby R.sup.6, R.sup.7, R.sup.8 and R.sup.9 are optionally substituted with one or two substituents, independently from each other, selected from OH, RO(CO)—, RR.sup.2N(CO)—, RR.sup.2NSO.sub.2— or 5-tetrazolyl; R is hydrogen or a C.sub.1-C.sub.4-alkyl group; or a stereoisomer, a tautomer, an N-oxide, a hydrate, a solvate, or a salt thereof, or a mixture consisting of the above.

3. Compounds of the general formula (I) according to claim 1, where A represents a group selected from: ##STR00112## wherein * indicates the point of attachment of said group with the rest of the molecule; X is a group selected from: ##STR00113## wherein * indicates the point of attachment of the group with the rest of the molecule and where the group is optionally substituted, one or two times, independently from each other, with a substituent selected from fluoro, chloro, methyl, trifluoromethyl or methoxy; R.sup.1 is propyl, which is optionally substituted with RO(CO)—; R.sup.2, R.sup.5 are, independently from each other, hydrogen or methyl; R.sup.3, R.sup.4 are, independently from each other, C.sub.1-C.sub.3-alkyl, and whereby R.sup.3 and R.sup.4 are optionally substituted with one or two substituents, independently from each other, selected from OH, RO(CO)—, RR.sup.2N(CO)—, RR.sup.2NSO.sub.2—, C.sub.1-C.sub.2-alkyl-(CO)(NH)SO.sub.2— or 5-tetrazolyl; R.sup.6, R.sup.7, R.sup.8, R.sup.9 are, independently from each other, C.sub.1-C.sub.4-alkyl, and whereby R.sup.6, R.sup.7, R.sup.8 and R.sup.9 are optionally substituted with one or two substituents, independently from each other, selected from OH, RO(CO)— or RR.sup.2N(CO)—; R is hydrogen or a C.sub.1-C.sub.4-alkyl group; or a stereoisomer, a tautomer, an N-oxide, a hydrate, a solvate, or a salt thereof, or a mixture consisting of the above.

4. Compounds of the general formula (I) according to claim 1, where A represents a group selected from: ##STR00114## wherein * indicates the point of attachment of said group with the rest of the molecule; X is a group selected from: ##STR00115## wherein * indicates the point of attachment of the group with the rest of the molecule and, R.sub.X is hydrogen or methyl, R.sup.Y is hydrogen, fluoro, chloro, methyl, trifluoromethyl or methoxy, R.sup.Z is hydrogen or methyl, wherein only one of R.sup.X, R.sup.Y and R.sup.Z is different from hydrogen and; R.sup.1 is —CH.sub.2—CH.sub.2—CH.sub.2—COOH; R.sup.2 is hydrogen or methyl; R.sup.3 is C.sub.1-C.sub.3-alkyl, which is optionally substituted with one substituent, selected from OH, HO(CO)—, H.sub.2N(CO)—, CH.sub.3—(CO)(NH)SO.sub.2— or 5-tetrazolyl; R.sup.4 is —CH.sub.2—CH.sub.2—COOH; R.sup.5 is hydrogen; R.sup.6 is C.sub.1-C.sub.4-alkyl, which is optionally substituted with one substituent, selected from OH, RO(CO)— or H.sub.2N(CO)—; R.sup.7 is methyl; R.sup.8 is —CH.sub.2—CH.sub.2—COOH; R.sup.9 is C.sub.2-C.sub.3-alkyl, which is substituted with one substituent, selected from OH or HO(CO)—; R is hydrogen or methyl; or a stereoisomer, a tautomer, an N-oxide, a hydrate, a solvate, or a salt thereof, or a mixture consisting of the above.

5. Compounds according to claim 1, selected from a group comprising the following compounds: tert-butyl N-{[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl]sulfonyl}-N-methyl-beta-alaninate N-{[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl]sulfonyl}-N-methyl-beta-alanine 17-(5-fluoropyridin-3-yl)-N-(3-hydroxypropyl)-N-methylestra-1(10),2,4,16-tetraene-3-sulfonamide tert-butyl N-methyl-N-{[17-(pyrimidin-5-yl)estra-1(10),2,4,16-tetraen-3-yl]sulfonyl}-beta-alaninate N-methyl-N-{[17-(pyrimidin-5-yl)estra-1(10),2,4,16-tetraen-3-yl]sulfonyl}-beta-alanine tert-butyl N-methyl-N-{[17-(6-methylpyridazin-4-yl)estra-1(10),2,4,16-tetraen-3-yl]sulfonyl}-beta-alaninate N-methyl-N-{[17-(6-methylpyridazin-4-yl)estra-1(10),2,4,16-tetraen-3-yl]sulfonyl}-beta-alanine 17-(5-fluoropyridin-3-yl)-3-(methylsulfonyl)estra-1(10),2,4,16-tetraene methyl 4-{[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl]sulfonyl}butanoate 4-{[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl]sulfonyl}butanoic acid methyl 4-{[17-(pyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl]sulfonyl}butanoate 4-{[17-(pyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl]sulfonyl}butanoic acid methyl 4-({17-[5-(trifluoromethyl)pyridin-3-yl]estra-1(10),2,4,16-tetraen-3-yl}sulfonyl) butanoate 4-({17-[5-(trifluoromethyl)pyridin-3-yl]estra-1(10),2,4,16-tetraen-3-yl}sulfonyl)butanoic acid 4-{[17-(6-methylpyridin-3-yl)estra-1,3,5(10),16-tetraen-3-yl]sulfonyl}butanoic acid 4-{[17-(5-methoxypyridin-3-yl)estra-1,3,5(10),16-tetraen-3-yl]sulfonyl}butanoic acid 4-{[17-(5-methylpyridin-3-yl)estra-1,3,5(10),16-tetraen-3-yl]sulfonyl}butanoic acid 4-{[17-(4-methylpyridin-3-yl)estra-1,3,5(10),16-tetraen-3-yl]sulfonyl}butanoic acid 4-{[17-(5-chloropyridin-3-yl)estra-1,3,5(10),16-tetraen-3-yl]sulfonyl}butanoic acid 4-{[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl]sulfonyl}butanamide 4-{[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl]sulfonyl}butan-1-ol tert-butyl 4-{[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl]amino}-4-oxobutanoate 4-{[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl]amino}-4-oxobutanoic acid tert-butyl 4-oxo-4-{[17-(pyrimidin-5-yl)estra-1(10),2,4,16-tetraen-3-yl]amino}butanoate 4-oxo-4-{[17-(pyrimidin-5-yl)estra-1(10),2,4,16-tetraen-3-yl]amino}butanoic acid N-[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl]butanediamide N-[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl]-4-hydroxybutanamide N-[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl]-3-sulfamoylpropanamide 3-(acetylsulfamoyl)-N-[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl]propanamide N-[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl]-N-methyl-3-sulfamoylpropanamide 3-(acetylsulfamoyl)-N-[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl]-N-methylpropanamide N-[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl]-3-(2H-tetrazol-5-yl)propanamide N-[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl]-N-methyl-3-(2H-tetrazol-5-yl)propanamide tert-butyl 4-{[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl](methyl)amino}-4-oxobutanoate 4-{[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl](methyl)amino}-4-oxobutanoic acid methyl 3-{[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl](methyl)sulfamoyl}propanoate methyl 3-{[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl]sulfamoyl}propanoate 3-{[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl](methyl)sulfamoyl}propanoic acid 3-{[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl]sulfamoyl}propanoic acid N-[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl]acetamide ethyl N-{[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl]carbamoyl}-beta-alaninate N-{[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl]carbamoyl}-beta-alanine methyl 4-{[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl](methyl)amino}butanoate 4-{[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl](methyl)amino}butanoic acid 4-{[17-(6-methylpyridazin-4-yl)estra-1(10),2,4,16-tetraen-3-yl]amino}-4-oxobutanoic acid 17-(5-fluoropyridin-3-yl)-3-(S-methylsulfonimidoyl)estra-1(10),2,4,16-tetraene and the stereoisomers, tautomers, N-oxides, hydrates, solvates or salts thereof, or a mixture consisting of the above.

6. Compound of the formula (I) as defined in claim 1 for treatment and/or prophylaxis of diseases.

7. Compound of the formula (I) as defined in claim 1 for use in a method for treatment and/or prophylaxis of endometriosis, of leiomyoma, of uterine bleeding disorders, of dysmenorrhoea, of prostate carcinoma, of prostate hyperplasia, of acne, of seborrhoea, of hair loss, of premature sexual maturity, of polycystic ovary syndrome, of breast cancer, of lung cancer, of endometrial carcinoma, of renal cell carcinoma, of bladder carcinoma, of non-Hodgkins lymphoma, of chronic obstructive pulmonary disease (COPD), of obesity, or of inflammation-related pain.

8. Use of a compound according to claim 1 for production of a medicament for treatment and/or prophylaxis of diseases.

9. Use of a compound as defined in claim 1 for production of a medicament for prophylaxis of endometriosis, of leiomyoma, of uterine bleeding disorders, of dysmenorrhoea, of prostate carcinoma, of prostate hyperplasia, of acne, of seborrhoea, of hair loss, of premature sexual maturity, of polycystic ovary syndrome, of breast cancer, of lung cancer, of endometrial carcinoma, of renal cell carcinoma, of bladder carcinoma, of non-Hodgkins lymphoma, of chronic obstructive pulmonary disease (COPD), of obesity or of inflammation-related pain.

10. Medicament comprising a compound as defined in claim 1 in combination with one or more further active ingredients, especially with selective oestrogen receptor modulators (SERMs), oestrogen receptor (ER) antagonists, aromatase inhibitors, 17-HSD1 inhibitors, steroid sulphatase (STS) inhibitors, GnRH agonists and antagonists, kisspeptin receptor (KISSR) antagonists, selective androgen receptor modulators (SARMs), androgens, 5-reductase inhibitors, C(17,20)-lyase inhibitors, selective progesterone receptor modulators (SPRMs), gestagens, antigestagens, oral contraceptives, inhibitors of mitogen-activated protein (MAP) kinases and inhibitors of the MAP kinases (Mkk3/6, Mek1/2, Erk1/2), inhibitors of the protein kinases B (PKBα/β/γ; Akt1/2/3), inhibitors of the phosphoinositide 3-kinases (PI3K), inhibitors of cyclin-dependent kinase (CDK1/2), inhibitors of the hypoxia-induced signalling pathway (HIF1alpha inhibitors, activators of prolylhydroxylases), histone deacetylase (HDAC) inhibitors, prostaglandin F receptor (FP) (PTGFR) antagonists and non-steroidal inflammation inhibitors (NSAIDs).

11. Medicament comprising a compound of the formula (I) as defined in claim 1 in combination with an inert, nontoxic, pharmaceutically suitable excipient.

12. Medicament according to claim 10 for treatment and prophylaxis of endometriosis, of uterine leiomyoma, of uterine bleeding disorders, of dysmenorrhoea, of prostate carcinoma, of prostate hyperplasia, of acne, of seborrhoea, of hair loss, of premature sexual maturity, of polycystic ovary syndrome, of breast cancer, of lung cancer, of endometrial carcinoma, of renal cell carcinoma, of bladder carcinoma, of non-Hodgkins lymphoma, of chronic obstructive pulmonary disease (COPD), of obesity or of inflammation-related pain.

13. Use of a compound of the general formula (I), according to claim 1 in the form of a pharmaceutical formulation for enteral, parenteral, vaginal, intrauterine or oral administration.

Description

EXAMPLES TYPE IC1

Example 1

tert-butyl N-{[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl]sulfonyl}-N-methyl-beta-alaninate

[0461] ##STR00061##

[0462] To a solution of 655 mg (1.07 mmol) of tert-butyl N-methyl-N-{[17-{[(trifluoromethyl)sulfonyl]oxy}estra-1(10),2,4,16-tetraen-3-yl]sulfonyl}-beta-alaninate (Intermediate 2-A) in 6.5 ml of dioxane 258 mg (1.83 mmol) of 5-fluoropyridine-3-boronic acid, 60.5 mg (0.08 mmol) of dichlorobis(triphenylphosphine)palladium(II) and 2.15 ml of 2M sodium carbonate solution were added. The reaction mixture was degassed and then stirred in a sealed tube for 2 hours at 90° C. The reaction mixture was allowed to cool to room temperature and partitioned between water and ethyl acetate. The organic phase was further washed with brine, dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by chromatography using a prepacked silica gel cartridge (mobile phase: cyclohexane/ethyl acetate, gradient 0% to 35%). The yield was 525 mg (83% of theory) of the title compound.

[0463] LC-MS (method A): R.sub.t=4.89 min; m/z=555.4 (M+H).sup.+

[0464] .sup.1H-NMR (300 MHz, CDCl.sub.3): δ [ppm]=1.07 (s, 3H), 1.45 (s, 9H), 1.48-1.61 (m, 1H), 1.67-1.86 (m, 4H), 1.96-2.10 (m, 1H), 2.04-2.05 (m, 2H), 2.41 (ddd, 3H), 2.54 (dd, 2H), 2.77 (s, 3H), 3.00 (dd, 2H), 3.29 (dd, 2H), 6.11 (dd, 1H), 7.37-7.45 (m, 2H), 7.50-7.57 (m, 2H), 8.36 (d, 1H), 8.48 (dd, 1H)

Example 2

N-{[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl]sulfonyl}-N-methyl-beta-alanine

[0465] ##STR00062##

[0466] 525 mg (0.94 mmol) of tert-butyl N-{[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl]sulfonyl}-N-methyl-beta-alaninate (Example 1) in 15.7 ml dichloromethane were treated with 3.64 ml (47.3 mmol) of trifluoroacetic acid. The mixture was stirred for 1 h at RT and concentrated under reduced pressure. The crude residue was purified by chromatography using a prepacked silica gel cartridge (mobile phase: dichloromethane/ethyl acetate:methanol: 30% triethylamine in water 7:3, gradient 0% to 70%). The yield was 326 mg (68% of theory) of the title compound.

[0467] LC-MS (method B): R.sub.t=5.28 min, m/z=499.2 (M+H).sup.+

[0468] .sup.1H-NMR (400 MHz, DMSO-d.sub.6): δ [ppm]=1.01 (s, 3H), 1.40-1.43 (m, 1H), 1.55-1.79 (m, 4H), 1.88-1.95 (m, 1H), 2.10-2.20 (m, 2H), 2.26-2.45 (m, 5H), 2.64 (s, 3H), 2.91-2.99 (m, 2H), 3.14 (dd, 2H), 6.26-6.28 (m, 1H), 7.43-7.53 (m, 3H), 7.66-7.71 (m, 1H), 8.43-8.51 (m, 2H), 11.2-13.4 (br. s, 1H)

Example 3

17-(5-fluoropyridin-3-yl)-N-(3-hydroxypropyl)-N-methylestra-1(10),2,4,16-tetraene-3-sulfonamide

[0469] ##STR00063##

[0470] 150 mg (0.30 mmol) of N-{[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl]sulfonyl}-N-methyl-beta-alanine (Example 2) in 1.2 ml tetrahydrofuran were treated at 0° C. with 50 μl (0.36 mmol) of triethylamine, followed by dropwise addition of 47 μl (0.36 mmol) of 2-methylpropyl carbonochloridate (CAS No.: 543-27-1). The reaction mixture was stirred at 0° C. for 45 min and then allowed to warm to room temperature for 15 minutes. The reaction mixture was cooled at 0° C., filtered, rinsed with tetrahydrofuran (200 μl) and the filtrate was cooled at 0° C. 13.6 mg (0.36 mmol) of sodium borohydride was added to the reaction mixture and after 5 minutes 300 μl of methanol. The reaction mixture was stirred at 0° C. for 45 minutes and then allowed to slowly warm to room temperature over 2 hours. Water was carefully added and stirring continued for 2 minutes. The reaction mixture was partitioned between water and ethyl acetate. The organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by chromatography on silica gel (mobile phase: dichloromethane/methanol, gradient 0% to 3%). The yield was 63 mg (43% of theory) of the title compound.

[0471] LC-MS (method B): R.sub.t=5.33 min; m/z=485.1 (M+H).sup.+

[0472] .sup.1H-NMR (400 MHz, DMSO-d.sub.6): δ [ppm]=1.01 (s, 3H), 1.56-1.77 (m, 6H), 1.88-1.97 (m, 1H), 2.09-2.20 (m, 2H), 2.26-2.45 (m, 4H), 2.62 (s, 3H), 2.95 (dd, 4H), 3.39 (dd, 2H), 4.43 (dd, 1H), 6.26-6.28 (m, 1H), 7.42-7.52 (m, 3H), 7.66-7.71 (m, 1H), 8.43 (d, 1H), 8.51 (t, 1H).

Example 4

tert-butyl N-methyl-N-{[17-(pyrimidin-5-yl)estra-1(10),2,4,16-tetraen-3-yl]sulfonyl}-beta-alaninate

[0473] ##STR00064##

[0474] Analogously to the preparation of Example 1, 268 mg (0.44 mmol) of tert-butyl N-methyl-N-{[17-{[(trifluoromethyl)sulfonyl]oxy}estra-1(10),2,4,16-tetraen-3-yl]sulfonyl}-beta-alaninate (Intermediate 2-A) were reacted with 93 mg (1.83 mmol) of pyrimidin-5-ylboronic acid and 24.7 mg (0.03 mmol) of dichlorobis(triphenylphosphine)palladium(II). The residue was purified by chromatography using a prepacked silica gel cartridge (mobile phase: cyclohexane/ethyl acetate, gradient 0% to 50%). The yield was 214 mg (86% of theory) of the title compound.

[0475] LC-MS (method A): R.sub.t=4.50 min; m/z=538.4 (M+H).sup.+

[0476] .sup.1H-NMR (300 MHz, CDCl.sub.3): δ [ppm]=1.07 (s, 3H), 1.26 (dd, 2H), 1.45 (s, 1H), 1.5-1.9 (m, 7H), 2.05 (s, 3H), 2.15-2.27 (m, 2H), 2.38-2.58 (m, 6H), 2.77 (s, 3H), 3.00 (dd, 2H), 3.29 (dd, 2H), 4.12 (q, 1H), 6.15 (dd, 1H), 7.42 (d, 1H), 7.51-7.57 (m, 2H), 8.76 (s, 2H), 9.10 (s, 1H).

Example 5

N-methyl-N-{[17-(pyrimidin-5-yl)estra-1(10),2,4,16-tetraen-3-yl]sulfonyl}-beta-alanine

[0477] ##STR00065##

[0478] Analogously to the preparation of Example 2, 213 mg (0.39 mmol) of tert-butyl N-methyl-N-{[17-(pyrimidin-5-yl)estra-1(10),2,4,16-tetraen-3-yl]sulfonyl}-beta-alaninate were treated with 1.52 ml (19.8 mmol) of trifluoroacetic acid. The crude residue was purified by preparative HPLC (Method E). The yield was 142 mg (74% of theory) of the title compound.

[0479] LC-MS (method B): R.sub.t=4.57 min; m/z=482.1 (M+H)+

[0480] .sup.1H-NMR (400 MHz, DMSO-d.sub.6): δ [ppm]=1.01 (s, 3H), 1.46-1.77 (m, 6H), 1.87-1.96 (m, 1H), 2.10-2.20 (m, 2H), 2.28-2.45 (m, 4H), 2.64 (s, 3H), 2.91-2.98 (m, 2H), 3.13 (dd, 2H), 6.31 (dd, 1H), 7.43-7.53 (m, 3H), 8.83 (s, 2H), 9.05 (s, 1H), 12.3 (br. s, 1H).

Example 6

tert-butyl N-methyl-N-{[17-(6-methylpyridazin-4-yl)estra-1(10),2,4,16-tetraen-3-yl]sulfonyl}-beta-alaninate

[0481] ##STR00066##

[0482] To a solution of 267 mg (0.44 mmol) of tert-butyl N-methyl-N-{[17-{[(trifluoromethyl)sulfonyl]oxy}estra-1(10),2,4,16-tetraen-3-yl]sulfonyl}-beta-alaninate (Intermediate 2-A) in 2.2 ml of toluene and 1.4 ml of ethanol 135 mg (0.61 mmol) of 3-methyl-5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-pyridazine, 37.2 mg (0.88 mmol) of lithium chloride, 24.7 mg (0.03 mmol) of dichlorobis(triphenylphosphine)palladium(II) and 549 μl of 2M sodium carbonate solution were added. The reaction mixture was degassed and then stirred in a sealed tube for 8 hours at 100° C. The reaction mixture was allowed to cool to room temperature and partitioned between water and ethyl acetate. The organic phase was further washed with brine, dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by chromatography using a prepacked silica gel cartridge (mobile phase: cyclohexane/ethyl acetate, gradient 0% to 70%). The yield was 130 mg (51% of theory) of the title compound.

[0483] LC-MS (method A): R.sub.t=4.03 min; m/z=552.3 (M+H).sup.+

[0484] .sup.1H-NMR (300 MHz, CDCl.sub.3): δ [ppm]=1.10 (s, 3H), 1.45 (s, 9H), 1.47-1.90 (m, 5H), 1.97-2.09 (m, 1H), 2.15-2.31 (m, 2H), 2.39-2.58 (m, 5H), 2.71 (s, 3H), 2.77 (s, 3H), 3.00 (dd, 2H), 3.29 (dd, 2H), 6.36 (dd, 1H), 7.25 (s, 1H), 7.42 (d, 1H), 7.51-7.58 (m, 2H), 9.08 (d, 1H).

Example 7

N-methyl-N-{[17-(6-methylpyridazin-4-yl)estra-1(10),2,4,16-tetraen-3-yl]sulfonyl}-beta-alanine

[0485] ##STR00067##

[0486] Analogously to the preparation of Example 2, 130 mg (0.23 mmol) of tert-butyl N-methyl-N-{[17-(6-methylpyridazin-4-yl)estra-1(10),2,4,16-tetraen-3-yl]sulfonyl}-beta-alaninate were treated with 908 μl (11.7 mmol) of trifluoroacetic acid. The crude residue was purified by preparative HPLC (Method E). The yield was 78 mg (66% of theory) of the title compound.

[0487] LC-MS (method B): R.sub.t=3.95 min; m/z=496.1 (M+H).sup.+

[0488] .sup.1H-NMR (400 MHz, DMSO-d.sub.6): δ [ppm]=1.03 (s, 3H), 1.44-1.78 (m, 6H), 1.87-1.96 (m, 1H), 2.16 (dd, 1H), 2.26-2.46 (m, 5H), 2.59 (s, 3H), 2.64 (s, 3H), 2.91-2.98 (m, 2H), 3.13 (dd, 2H), 6.56-6.57 (m, 1H), 7.43-7.51 (m, 4H), 9.11 (d, 1H), 12.5 (br. s, 1H).

Examples Type IC2

Example 8

17-(5-fluoropyridin-3-yl)-3-(methylsulfonyl)estra-1(10),2,4,16-tetraene

[0489] ##STR00068##

[0490] Analogously to the preparation of Example 1, 65 mg (0.14 mmol) of 3-(methylsulfonyl)estra-1(10),2,4,16-tetraen-17-yl trifluoromethanesulfonate (Intermediate 5-B) were treated with 21.7 mg (0.15 mmol) of 5-fluoropyridine-3-boronic acid (CAS No.: 872041-86-6) and 7.85 mg (0.01 mmol) of dichlorobis(triphenylphosphine)palladium(II) (CAS No.: 13965-03-2). The residue was purified using a prepacked silica gel cartridge (mobile phase: cyclohexane/ethyl acetate, gradient 0% to 35%). The yield was 53.4 mg (91% of theory) of the title compound.

[0491] LC-MS (method B): R.sub.t=5.43 min; m/z=412.1 (M+H).sup.+

[0492] .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm]=1.01 (s, 3H), 1.45-1.77 (m, 6H), 1.89-1.97 (m, 1H), 2.09-2.21 (m, 2H), 2.26-2.45 (m, 2H), 2.92-2.98 (m, 2H), 3.14 (s, 3H), 6.28 (dd, 1H), 7.53 (d, 1H), 7.60-7.65 (m, 2H), 7.67-7.71 (m, 1H), 8.44 (d, 1H), 8.50 (t, 1H).

Example 9

methyl 4-{[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl]sulfonyl}butanoate

[0493] ##STR00069##

[0494] Analogously to the preparation of Example 1, 2.1 g (3.35 mmol) of methyl 4-{[17-{[(trifluoromethyl)sulfonyl]oxy}estra-1(10),2,4,16-tetraen-3-yl]sulfonyl}butanoate (Intermediate 5-A) were treated with 804 mg (5.70 mmol) of 5-fluoropyridine-3-boronic acid (CAS No.: 872041-86-6) and 188 mg (0.26 mmol) of dichlorobis(triphenylphosphine)palladium(II) (CAS No.: 13965-03-2). The residue was purified using a prepacked silica gel cartridge (mobile phase: cyclohexane/ethyl acetate, gradient 0% to 45%). The yield was 1.23 g (73% of theory) of the title compound.

[0495] LC-MS (method B): R.sub.t=5.61 min; m/z=498.2 (M+H).sup.+

[0496] .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm]=1.01 (s, 3H), 1.37-1.54 (m, 1H), 1.58-1.81 (m, 6H), 1.89-1.96 (m, 1H), 2.09-2.21 (m, 2H), 2.28-2.36 (m, 1H), 2.37-2.46 (m, 4H), 2.94-2.99 (m, 2H), 3.22-3.31 (m, 2H), 3.54 (s, 3H), 6.28 (dd, 1H), 7.53-7.61 (m, 3H), 7.67-7.72 (m, 1H), 8.44 (d, 1H), 8.50 (t, 1H).

Example 10

4-{[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl]sulfonyl}butanoic acid

[0497] ##STR00070##

[0498] 300 mg (0.60 mmol) of methyl 4-{[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl]sulfonyl}butanoate (Example 9) in 4.2 ml tetrahydrofuran were treated with 603 μl (1.20 mmol) of 2N lithium hydroxide solution. The mixture was stirred for 16 hours. The reaction mixture was acidified to pH 4 by addition of 1N hydrochloric acid solution and the reaction mixture was concentrated in vacuo. A part of the crude residue was purified by preparative HPLC (Method E). The yield was 66.3 mg of the title compound.

[0499] LC-MS (method B): R.sub.t=5.03 min; m/z=484.1 (M+H).sup.+

[0500] 1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm]=1.01 (s, 3H), 1.41-1.43 (m, 1H), 1.57-1.78 (m, 5H), 1.87-1.97 (m, 1H), 2.08-2.20 (m, 2H), 2.27-2.45 (m, 6H), 2.94-2.99 (m, 2H), 3.18-3.28 (m, 2H), 6.28 (dd, 1H), 7.52-7.60 (m, 3H), 7.66-7.71 (m, 1H), 8.44 (d, 1H), 8.50 (t, 1H, J=1.7 Hz), 12.2 (br. s, 1H).

Example 11

methyl 4-{[17-(pyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl]sulfonyl}butanoate

[0501] ##STR00071##

[0502] Analogously to the preparation of Example 9, 90 mg (0.16 mmol) of methyl 4-{[17-{[(trifluoromethyl)sulfonyl]oxy}estra-1(10),2,4,16-tetraen-3-yl]sulfonyl}butanoate (Intermediate 5-A) were treated with 34.1 mg (0.27 mmol) of pyridin-3-ylboronic acid (CAS No.: 1692-25-7) and 9.2 mg (0.013 mmol) of dichlorobis(triphenylphosphine)palladium(II) (CAS No.: 13965-03-2). The residue was purified using a prepacked silica gel cartridge (mobile phase: dichloromethane/ethyl acetate, gradient 0% to 40%). The yield was 41 mg (52% of theory) of the title compound.

[0503] LC-MS (method C): R.sub.t=3.48 min; m/z=498.1 (M+H).sup.+

[0504] .sup.1H NMR (300 MHz, CDCl.sub.3): δ [ppm]=1.07 (s, 3H), 1.46-1.63 (m, 1H), 1.64-1.91 (m, 5H), 2.00-2.24 (m, 3H), 2.38-2.51 (m, 6H), 3.01 (dd, 2H), 3.13-3.20 (m, 2H), 3.67 (s, 3H), 6.04 (dd, 1H), 7.22-7.26 (m, 1H), 7.47 (d, 1H), 7.61-7.71 (m, 3H), 8.49 (d, 1H), 8.64-8.67 (m, 1H).

Example 12

4-{[17-(pyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl]sulfonyl}butanoic acid

[0505] ##STR00072##

[0506] Analogously to the preparation of Example 10, 41 mg (0.085 mmol) of methyl 4-{[17-(pyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl]sulfonyl}butanoate were treated with 85 μl (0.17 mmol) of 2N lithium hydroxide solution. The crude residue was purified by chromatography using a prepacked C18 cartridge (mobile phase: acetonitrile/water 0.1% formic acid buffer, gradient 10% to 60%). The yield was 34.0 mg (84% of theory) of the title compound.

[0507] LC-MS (method B): R.sub.t=3.55 min; m/z=466.1 (M+H).sup.+

[0508] .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm]=1.00 (s, 3H), 1.41-1.53 (m, 1H), 1.57-1.80 (m, 6H), 1.89-1.96 (m, 1H), 2.07-2.17 (m, 2H), 2.27-2.33 (m, 3H), 2.33-2.45 (m, 2H), 2.92-2.99 (m, 2H), 3.10-3.19 (m, 2H), 6.12-6.14 (m, 1H), 7.33 (dd, 1H), 7.52-7.60 (m, 3H), 7.76-7.80 (m, 1H), 8.43 (dd, 1H), 8.60 (d, 1H).

Example 13

methyl 4-({17-[5-(trifluoromethyl)pyridin-3-yl]estra-1(10),2,4,16-tetraen-3-yl}sulfonyl)butanoate

[0509] ##STR00073##

[0510] Analogously to the preparation of Example 9, 90 mg (0.16 mmol) of methyl 4-{[17-{[(trifluoromethyl)sulfonyl]oxy}estra-1(10),2,4,16-tetraen-3-yl]sulfonyl}butanoate (Intermediate 5-A) were treated with 53.0 mg (0.27 mmol) of [5-(trifluoromethyl)pyridin-3-yl]boronic acid (CAS No.: 947533-51-9) and 9.2 mg (0.013 mmol) of dichlorobis(triphenylphosphine)palladium(II) (CAS No.: 13965-03-2). The residue was purified using a prepacked silica gel cartridge (mobile phase: cyclohexane/ethyl acetate, gradient 0% to 40%). The yield was 66 mg (73% of theory) of the title compound.

[0511] LC-MS (method C): R.sub.t=4.35 min; m/z=548.1 (M+H).sup.+

[0512] .sup.1H NMR (300 MHz, CDCl.sub.3): δ [ppm]=1.09 (s, 3H), 1.53-1.59 (m, 2H), 1.69-1.78 (m, 2H), 1.79-1.93 (m, 1H), 1.98-2.11 (m, 3H), 2.15-2.28 (m, 2H), 2.40-2.52 (m, 5H), 3.02 (dd, 2H), 3.13-3.20 (m, 2H), 3.67 (s, 3H), 6.14-6.18 (m, 1H), 7.47 (d, 1H), 7.62-7.69 (m, 2H), 7.89 (s, 1H), 8.77 (s, 1H), 8.82-8.83 (m, 1H).

Example 14

4-({17-[5-(trifluoromethyl)pyridin-3-yl]estra-1(10),2,4,16-tetraen-3-yl}sulfonyl)butanoic acid

[0513] ##STR00074##

[0514] Analogously to the preparation of Example 10, 66 mg (0.12 mmol) of methyl 4-({17-[5-(trifluoromethyl)pyridin-3-yl]estra-1(10),2,4,16-tetraen-3-yl}sulfonyl)butanoate (Example 10) were treated with 121 μl (0.24 mmol) of 2N lithium hydroxide solution. The crude residue was purified by chromatography using a prepacked C18 cartridge (mobile phase: acetonitrile/water 0.1% formic acid buffer, gradient 20% to 80%). The yield was 55 mg of the title compound.

[0515] LC-MS (method B): R.sub.t=5.47 min, m/z=534.1 (M+H).sup.+

[0516] .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm]=1.02 (s, 3H), 1.46-1.79 (m, 6H), 1.91-1.98 (m, 1H), 2.11-2.19 (m, 2H), 2.27-2.44 (m, 6H), 2.93-3.00 (m, 2H), 3.17-3.24 (m, 2H), 6.35-6.38 (m, 1H), 7.52-7.61 (m, 3H), 8.05 (s, 1H), 8.84 (d, 1H), 8.91 (d, 1H), 12.12 (br. s, 1H).

Example 15

4-{[17-(6-methylpyridin-3-yl)estra-1,3,5(10),16-tetraen-3-yl]sulfonyl}butanoic acid

[0517] ##STR00075##

[0518] To 95 mg (0.17 mmol) methyl 4-{[17-{[(trifluoromethyl)sulfonyl]oxy}estra-1(10),2,4,16-tetraen-3-yl]sulfonyl}butanoate (Intermediate 5-A) in 2 ml dioxane 0.26 ml 2M aqueous sodium carbonate solution, 35 mg (0.26 mmol, 1.5 eq.) (6-methylpyridin-3-yl)boronic acid and 12 mg dichlorobis(triphenylphosphine)palladium(II) (CAS No.: 13965-03-2) were added and the mixture was heated in a closed vessel at 90° C. Then 0.4 ml 2M aqeous sodium hydroxide solution were added and the mixture was stirred for 17.5 h at 50° C. The mixture was diluted with water, acidified by aqueous citric acid to pH 4, extracted three times with ethyl acetate, evaporated and purified by HPLC (method G) resulting in 36 mg (43% yield) of the title compound.

[0519] LC-MS (method F): R.sub.t=0.93 min, m/z=479.21.

[0520] .sup.1H NMR (400 MHz, DMSO-d6): δ [ppm]=1.02 (s, 3H), 1.43-1.83 (m), 1.91-2.00 (m, 1H), 2.09-2.21 (m, 2H), 2.26-2.49 (m), 2.94-3.04 (m, 2H), 3.24-3.32 (m), 6.06-6.11 (m, 1H), 7.22 (d, 1H), 7.55-7.65 (m, 3H), 7.70 (dd, 1H), 8.49 (d, 1H), 12.20 (br. s., 1H).

Example 16

4-{[17-(5-methoxypyridin-3-yl)estra-1,3,5(10),16-tetraen-3-yl]sulfonyl}butanoic acid

[0521] ##STR00076##

[0522] Analogously to Example 15 reaction of 95 mg (0.17 mmol) methyl 4-{[17-{[(trifluoromethyl)sulfonyl]oxy}estra-1(10),2,4,16-tetraen-3-yl]sulfonyl}butanoate (Intermediate 5-A) with 40 mg (5-methoxypyridin-3-yl)boronic acid afforded 33 mg of the title compound.

[0523] LC-MS (method F): R.sub.t=1.12 min, m/z=495.21.

[0524] .sup.1H NMR (400 MHz, DMSO-d6): δ [ppm]=1.04 (s, 3H), 1.44-1.84 (m), 1.91-2.01 (m, 1H), 2.10-2.23 (m, 2H), 2.28-2.49 (m), 2.93-3.04 (m, 2H), 3.25-3.31 (m), 3.86 (s, 3H), 6.18-6.23 (m, 1H), 7.28-7.31 (m, 1H), 7.54-7.65 (m, 3H), 8.20 (d, 1H), 8.25 (d, 1H), 12.21 (br. s., 1H).

Example 17

4-{[17-(5-methylpyridin-3-yl)estra-1,3,5(10),16-tetraen-3-yl]sulfonyl}butanoic acid

[0525] ##STR00077##

[0526] Analogously to Example 15 reaction of 95 mg (0.17 mmol) methyl 4-{[17-{[(trifluoromethyl)sulfonyl]oxy}estra-1(10),2,4,16-tetraen-3-yl]sulfonyl}butanoate (Intermediate 5-A) with 35 mg (5-methylpyridin-3-yl)boronic acid afforded 18 mg of the title compound.

[0527] LC-MS (method F): R.sub.t=0.99 min, m/z=479.21.

[0528] .sup.1H NMR (400 MHz, DMSO-d6): δ [ppm]=1.04 (s, 3H), 1.44-1.84 (m), 1.90-2.00 (m, 1H), 2.10-2.23 (m, 2H), 2.25-2.49 (m), 2.92-3.04 (m, 2H), 3.22-3.32 (m), 6.14 (br. s., 1H), 7.53-7.67 (m, 4H), 8.29-8.33 (m, 1H), 8.43 (d, 1H), 12.21 (br. s., 1H).

Example 18

4-{[17-(4-methylpyridin-3-yl)estra-1,3,5(10),16-tetraen-3-yl]sulfonyl}butanoic acid

[0529] ##STR00078##

[0530] Analogously to Example 15 reaction of 95 mg (0.17 mmol) methyl 4-{[17-{[(trifluoromethyl)sulfonyl]oxy}estra-1(10),2,4,16-tetraen-3-yl]sulfonyl}butanoate (Intermediate 5-A) with 35 mg (4-methylpyridin-3-yl)boronic acid afforded 22 mg of the title compound.

[0531] LC-MS (method F): R.sub.t=0.93 min, m/z=479.21.

[0532] .sup.1H NMR (400 MHz, DMSO-d6): δ [ppm]=0.91 (s, 3H), 1.45-1.80 (m), 1.80-1.90 (m, 1H), 1.93-2.04 (m, 1H), 2.20 (dd, 1H), 2.29 (s, 3H), 2.31-2.47 (m), 2.53-2.58 (m, 1H), 2.95-3.05 (m, 2H), 3.24-3.31 (m), 5.77 (d, 1H), 7.28 (d, 1H), 7.52-7.65 (m, 3H), 8.26 (s, 1H), 8.33 (d, 1H), 12.21 (br. s., 1H).

Example 19

4-{[17-(5-chloropyridin-3-yl)estra-1,3,5(10),16-tetraen-3-yl]sulfonyl}butanoic acid

[0533] ##STR00079##

[0534] Analogously to Example 15 reaction of 95 mg (0.17 mmol) methyl 4-{[17-{[(trifluoromethyl)sulfonyl]oxy}estra-1(10),2,4,16-tetraen-3-yl]sulfonyl}butanoate (Intermediate 5-A) with 41 mg (5-chloropyridin-3-yl)boronic acid afforded 39 mg of the title compound.

[0535] LC-MS (method F): R.sub.t=1.41 min, m/z=499.16.

[0536] .sup.1H NMR (400 MHz, DMSO-d6): δ [ppm]=1.04 (s, 3H), 1.47-1.84 (m), 1.91-2.01 (m, 1H), 2.10-2.25 (m, 2H), 2.27-2.49 (m), 2.92-3.04 (m, 2H), 3.21-3.32 (m), 6.29-6.34 (m, 1H), 7.54-7.65 (m, 3H), 7.90 (t, 1H), 8.53 (d, 1H), 8.61 (d, 1H), 12.22 (br. s., 1H).

Example 20

4-{[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl]sulfonyl}butanamide

[0537] ##STR00080##

[0538] To a solution of 248 mg (0.51 mmol) of 4-{[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl]sulfonyl}butanoic acid (Example 10) in N,N-dimethylformamide (5.2 ml) 124 μl (1.12 mmol) of 4-methylmorpholine, 214 mg (0.56 mmol) of HATU and after 10 min, 366 μl (2.564 mmol) of 7N ammonia in methanol were added. The mixture was stirred for 2 hours. It was then partially concentrated under reduced pressure and the residue was partitioned between water and ethyl acetate.

[0539] The organic phase was further washed with brine, dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by HPLC (mobile phase: acetonitrile/water 0.1% formic acid buffer, gradient 10% to 95%). The yield was 137 mg (55% of theory) of the title compound.

[0540] LC-MS (method B): R.sub.t=4.71 min; m/z=483.2 (M+H).sup.+

[0541] .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm]=1.01 (s, 3H), 1.41-1.53 (m, 1H), 1.56-1.78 (m, 6H), 1.89-1.96 (m, 1H), 2.09-2.18 (m, 4H), 2.27-2.45 (m, 3H), 2.93-2.99 (m, 2H), 3.19-3.25 (m, 2H), 6.26-6.28 (m, 1H), 6.70-6.76 (m, 1H), 7.25-7.28 (m, 1H), 7.52-7.61 (m, 3H), 7.67-7.71 (m, 1H), 8.44 (d, 1H), 8.50 (t, 1H).

Example 21

4-{[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl]sulfonyl}butan-1-ol

[0542] ##STR00081##

[0543] To a solution of 200 mg (0.40 mmol) of methyl 4-{[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl]sulfonyl}butanoate (Example 9) in 6.7 ml of dichloromethane under argon at −78° C. 1.2 ml (1.20 mmol) of diisobutylaluminum hydride (1M in dichloromethane) was added dropwise over 5 minutes. After 10 minutes at −78° C. the reaction mixture was allowed to warm at −20° C. and stirred for 20 minutes and then allowed to warm to RT and stirred for 10 minutes. Addition of 500 μl of water dropwise and stirring for 10 minutes, then addition of 520 mg of sodium hydrogen carbonate and stirring for 30 minutes followed by sodium sulfate and stirring for 10 minutes, filtration and concentration in vacuo. The residue was purified by chromatography using a prepacked silica gel cartridge (mobile phase: acetonitrile/ethyl acetate, gradient 0% to 30%). The yield was 134 mg (69% of theory) of the title compound.

[0544] LC-MS (method B): R.sub.t=5.06 min; m/z=470.2 (M+H).sup.+

[0545] .sup.1H-NMR (400 MHz, DMSO-d.sub.6): δ [ppm]=1.01 (s, 3H), 1.40-1.77 (m, 9H), 1.87-1.97 (m, 1H), 2.09-2.20 (m, 2H), 2.27-2.45 (m, 3H), 2.92-2.99 (m, 2H), 3.18-3.25 (m, 4H), 4.40 (dd, 1H), 6.28 (dd, 1H), 7.51-7.61 (m, 3H), 7.66-7.71 (m, 1H), 8.44 (d, 1H), 8.50 (t, 1H).

Examples Type IC3

Example 22

tert-butyl 4-{[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl]amino}-4-oxobutanoate

[0546] ##STR00082##

[0547] To a solution of 1.41 g (2.63 mmol) of tert-butyl 4-{[17-iodoestra-1(10),2,4,16-tetraen-3-yl]amino}-4-oxobutanoate (Intermediate 8-A) in 6.8 ml of toluene and 4.4 ml of ethanol 519 mg (3.68 mmol) of 5-fluoropyridine-3-boronic acid (CAS No.: 872041-86-6), 223 mg (5.26 mmol) of lithium chloride, 243 mg (0.21 mmol) of tetrakis(triphenylphosphine)palladium(0) and 3.3 ml of 2M sodium carbonate solution were added. The reaction mixture was degassed and then stirred in a sealed tube for 3 hours at 100° C. The reaction mixture was allowed to cool to RT and partitioned between water and ethyl acetate. The organic phase was further washed with brine, dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by chromatography using a prepacked silica gel cartridge (mobile phase: dichloromethane/diethyl ether, gradient 0% to 60%). The yield was 457 mg (34% of theory) of the title compound.

[0548] LC-MS (method D): R.sub.t=4.65 min; m/z=505.3 (M+H).sup.+

[0549] .sup.1H-NMR (400 MHz, CDCl.sub.3): δ [ppm]=1.03 (s, 3H), 1.22-1.29 (m, 1H), 1.45 (s, 8H), 1.46-1.54 (m, 1H), 1.60-1.69 (m, 3H), 1.76-1.85 (m, 1H), 1.96 (ddd, 1H), 2.10-2.19 (m, 2H), 2.27-2.41 (m, 3H), 2.54-2.68 (m, 4H), 2.87-2.94 (m, 2H), 6.08 (dd, 1H), 7.19 (s, 2H), 7.31 (s, 1H), 7.36-7.41 (m, 1H), 7.56 (s, 1H), 8.33 (d, 1H), 8.46 (dd, 1H)

Example 23

4-{[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl]amino}-4-oxobutanoic acid

[0550] ##STR00083##

[0551] To a solution of 178 mg (0.35 mmol) of tert-butyl 4-{[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl]amino}-4-oxobutanoate (Example 22) in 7.4 ml of dichloromethane 1.77 ml (22.9 mmol) of trifluoroacetic acid was added. The mixture was stirred at RT for 1 hour and concentrated under reduced pressure. 69 mg of the residue were purified by preparative HPLC (Method E). The yield was 11 mg (20% of theory) of the title compound.

[0552] LC-MS (method B): R.sub.t=4.97 min; m/z=449.2 (M+H).sup.+

[0553] .sup.1H-NMR (400 MHz, DMSO-d.sub.6): δ [ppm]=1.00 (s, 3H), 1.36-1.62 (m, 4H), 1.67-1.77 (m, 1H), 1.84-1.92 (m, 1H), 2.08-2.17 (m, 2H), 2.20-2.39 (m, 3H), 2.75-2.84 (m, 2H), 3.16-3.44 (m, 4H), 6.25-6.27 (m, 1H), 7.14 (d, 1H), 7.26 (dd, 1H), 7.30-7.32 (m, 1H), 7.66-7.71 (m, 1H), 8.43 (d, 1H), 8.49 (dd, 1H), 9.80-9.85 (m, 1H), 12.10 (br. s, 1H).

Example 24

tert-butyl 4-oxo-4-{[17-(pyrimidin-5-yl)estra-1(10),2,4,16-tetraen-3-yl]amino}butanoate

[0554] ##STR00084##

[0555] To a solution of 250 mg (0.46 mmol) of tert-butyl 4-{[17-iodoestra-1(10),2,4,16-tetraen-3-yl]amino}-4-oxobutanoate (Intermediate 8-A) in 2.8 ml of dioxane 101 mg (0.81 mmol) of pyrimidin-5-ylboronic acid (CAS No.: 109299-78-7), 29.5 mg (0.04 mmol) of dichlorobis(triphenylphosphine)palladium(II) (CAS No.: 13965-03-2) and 934 μl of 2M sodium carbonate solution were added. The reaction mixture was degassed and then stirred in a sealed tube for 18 hours at 90° C. The reaction mixture was allowed to cool to RT and partitioned between water and ethyl acetate. The organic phase was further washed with brine, dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by chromatography using a prepacked silica gel cartridge (mobile phase: cyclohexane/ethyl acetate, gradient 0% to 50%). The yield was 58 mg (25% of theory) of the title compound.

[0556] LC-MS (method A): R.sub.t=4.11 min; m/z=488.2 (M+H).sup.+

[0557] .sup.1H NMR (300 MHz, CDCl.sub.3): δ [ppm]=0.81-0.95 (m, 1H), 1.45 (s, 9H), 1.50-1.90 (m, 5H), 1.91-2.03 (m, 1H), 2.11-2.24 (m, 2H), 2.28-2.47 (m, 3H), 2.54-2.71 (m, 4H), 2.88-2.97 (m, 2H), 6.13 (dd, 1H), 7.21 (s, 2H), 7.33 (s, 1H), 7.57 (s, 1H), 8.76 (s, 2H), 9.09 (s, 1H).

Example 25

4-oxo-4-{[17-(pyrimidin-5-yl)estra-1(10),2,4,16-tetraen-3-yl]amino}butanoic acid

[0558] ##STR00085##

[0559] Analogously to the Example 23, 58 mg (0.12 mmol) of tert-butyl 4-oxo-4-{[17-(pyrimidin-5-yl)estra-1(10),2,4,16-tetraen-3-yl]amino}butanoate (Example 24) were treated with 0.45 ml (5.94 mmol) of trifluoroacetic acid. The residue was diluted with ethyl acetate and saturated sodium hydrogen carbonate solution was added. The mixture was extracted 4 times with dichloromethane/methanol 9:1. The organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated in vacuo. The yield was 39.8 mg (77% of theory) of the title compound.

[0560] LC-MS (method B): R.sub.t=4.21 min; m/z=432.2 (M+H).sup.+

[0561] .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm]=0.78-0.85 (m, 1H), 0.99 (s, 3H), 1.12-1.63 (m, 6H), 1.68-1.77 (m, 1H), 1.86-1.96 (m, 1H), 2.08-2.38 (m, 7H), 2.63-2.84 (m, 2H), 6.30 (dd, 1H), 7.12 (d, 1H), 7.22-7.28 (m, 2H), 8.82 (s, 2H), 9.04 (s, 1H), 11.52 (br. s, 1H).

Example 26

N-[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl]butanediamide

[0562] ##STR00086##

[0563] To a solution of 210 mg (0.28 mmol) of 4-{[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl]amino}-4-oxobutanoic acid (Example 23) in 4.2 ml of N,N-dimethylformamide 68 μl (0.61 mmol) of 4-methylmorpholine, 117 mg (0.30 mmol) of 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxid hexafluorophosphate were added and after 5 minutes, 200 μl (1.40 mmol) of 7N ammonia in methanol. The mixture was stirred for 2 hours at RT. It was then partially concentrated under reduced pressure and the residue was partitioned between water and ethyl acetate. The organic phase was further washed with brine, dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by chromatography using a prepacked reverse phase cartridge (mobile phase: acetonitrile/water 0.1% formic acid buffer, gradient 20% to 95%). The yield was 23 mg (18% of theory) of the title compound.

[0564] LC-MS (method B): R.sub.t=4.67 min; m/z=448.2 (M+H).sup.+

[0565] .sup.1H-NMR (400 MHz, DMSO-d.sub.6): δ [ppm]=1.00 (s, 3H), 1.36-1.62 (m, 4H), 1.67-1.77 (m, 1H), 1.84-1.92 (m, 1H), 2.04-2.36 (m, 6H), 2.43-2.50 (m, 5H), 2.72-2.84 (m, 2H), 6.25-6.27 (m, 1H), 6.68-6.74 (m, 1H), 7.14 (d, 1H), 7.26 (dd, 2H), 7.31 (s, 1H), 7.65-7.70 (m, 1H), 8.43 (d, 1H), 8.49 (dd, 1H), 9.72 (s, 1H).

Example 27

N-[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl]-4-hydroxybutanamide

[0566] ##STR00087##

[0567] To a solution of 70 mg (0.20 mmol) of 17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-amine (Intermediate 10-A) in 1.8 ml of N,N-dimethylformamide under argon 30.4 mg (0.24 mmol) of 4-hydroxybutyric acid, sodium salt (CAS No.: 502-85-2), 27 μl (0.24 mmol) of 4-methylmorpholine and 91.6 mg (0.24 mmol) of 1-[bis-(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxid hexafluorophosphate were added. The mixture was stirred at RT for 2 hours. Then further 30.4 mg (0.24 mmol) of 4-hydroxybutyric acid, sodium salt, 27 μl (0.24 mmol) of 4-methylmorpholine and 91.6 mg (0.24 mmol) of 1-[bis-(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxid hexafluorophosphate were added and stirring continued for another 16 hours at RT. The reaction mixture was partitioned between water and ethyl acetate. The organic phase was further washed with brine, dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by chromatography using a prepacked silica gel cartridge (mobile phase: dichloromethane/methanol, gradient 0% to 5%) and then by chromatography using a prepacked reverse phase cartridge (mobile phase: acetonitrile/water 0.1% formic acid buffer, gradient 40% to 98%). The yield was 25 mg (28% of theory) of the title compound.

[0568] LC-MS (method B): R.sub.t=4.89 min; m/z=435.2 (M+H).sup.+

[0569] .sup.1H-NMR (400 MHz, CD.sub.3OD): δ [ppm]=1.08 (s, 3H), 1.42-1.54 (m, 1H), 1.61-1.72 (m, 3H), 1.79-2.02 (m, 4H), 2.15-2.23 (m, 2H), 2.39-2.47 (m, 6H), 2.86-2.93 (m, 2H), 3.60 (dd, 2H), 6.20 (dd, 1H), 7.18-7.27 (m, 3H), 7.61-7.66 (m, 1H), 8.32 (d, 1H), 8.44 (dd, 1H).

Example 28

N-[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl]-3-sulfamoylpropanamide

[0570] ##STR00088##

[0571] Analogously to the preparation of Intermediate 6-A, 155 mg (0.445 mmol) of 17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-amine (Intermediate 10-A) were treated with 372 mg (0.98 mmol) of 1-[bis-(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxid hexafluorophosphate, 150 mg (0.98 mmol) of 3-sulfamoylpropanoic acid (CAS No.: 15441-10-8) and 108 μl (0.98 mmol) of 4-methylmorpholine. The residue was purified by chromatography using a prepacked silica gel cartridge (mobile phase: dichloromethane/methanol, gradient 0% to 6%). The yield was 187 mg (86% of theory) of the title compound.

[0572] LC-MS (method A): R.sub.t=3.63 min; m/z=484.2 (M+H).sup.+

[0573] .sup.1H NMR (300 MHz, CDCl.sub.3): δ [ppm]=1.00 (s, 3H), 1.38-1.46 (m, 1H), 1.52-1.68 (m, 3H), 1.72-1.78 (m, 1H), 1.87-1.94 (m, 1H), 2.05-2.18 (m, 2H), 2.19-2.49 (m, 3H), 2.77-2.95 (m, 4H), 3.50-3.61 (m, 2H), 5.08-5.33 (m, 2H), 6.08 (s, 1H), 7.14-7.21 (m, 2H), 7.22-7.27 (m, 1H), 7.38 (d, 1H), 7.77-7.93 (m, 1H), 8.33 (d, 1H), 8.45 (s, 1H).

Example 29

3-(acetylsulfamoyl)-N-[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl]propanamide

[0574] ##STR00089##

[0575] To a solution of 167 mg (0.34 mmol) of N-[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl]-3-sulfamoylpropanamide (Example 28) in dichloromethane (8.5 ml) were added 35 μl (0.60 mmol) of acetic acid, 116 mg (0.60 mmol) of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and 73.8 mg (0.60 mmol) of 4-dimethylaminopyridine. The reaction was repeated using 20 mg N-[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl]-3-sulfamoylpropanamide. The combined reaction mixtures were diluted with dichloromethane and washed with brine, the organic phase was dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by reverse phase C18 chromatography (mobile phase: acetonitrile/water 0.1% formic acid buffer, gradient 10% to 95%). The yield was 126 mg of the title compound.

[0576] LC-MS (method B): R.sub.t=4.99 min; m/z=526.1 (M+H).sup.+

[0577] .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm]=1.00 (s, 3H), 1.37-1.62 (m, 4H), 1.68-1.77 (m, 1H), 1.84-1.92 (m, 1H), 1.96 (s, 3H), 2.08-2.40 (m, 5H), 2.70-2.85 (m, 4H), 3.61 (t, 2H), 6.25-6.27 (m, 1H), 7.17 (d, 1H), 7.24-7.30 (m, 2H), 7.66-7.70 (m, 1H), 8.44 (d, 1H), 8.48-8.51 (m, 1H), 9.92 (s, 1H), 11.63 (br. s., 1H).

Example 30

N-[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl]-N-methyl-3-sulfamoylpropanamide

[0578] ##STR00090##

[0579] Analogously to the preparation of Intermediate 6-A, 230 mg (0.634 mmol) of 17-(5-fluoropyridin-3-yl)-N-methylestra-1(10),2,4,16-tetraen-3-amine (Intermediate 11-A) were treated with 531 mg (1.39 mmol) of 1-[bis-(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxid hexafluorophosphate, 214 mg (1.39 mmol) of 3-sulfamoylpropanoic acid (CAS No.: 15441-10-8) and 153 μl (1.39 mmol) of 4-methylmorpholine. The residue was purified by chromatography using a prepacked silica gel cartridge (mobile phase: dichloromethane/methanol, gradient 0% to 5%). The yield was 246 mg (77% of theory) of the title compound.

[0580] LC-MS (method C): R.sub.t=3.46 min; m/z=498.0 (M+H).sup.+

[0581] .sup.1H NMR (400 MHz, CDCl.sub.3): δ [ppm]=1.09 (s, 3H), 1.46-1.60 (m, 1H), 1.63-1.89 (m, 4H), 1.98-2.05 (m, 1H), 2.14-2.24 (m, 2H), 2.36-2.47 (m, 3H), 2.67 (t, 2H), 2.91-2.98 (m, 2H), 3.26 (s, 3H), 3.45 (t, 2H), 4.90 (s, 2H), 6.11 (dd, 1H), 6.92-6.99 (m, 2H), 7.33 (d, 1H), 7.38-7.43 (m, 1H), 8.35 (d, 1H), 8.48 (dd, 1H).

Example 31

3-(acetylsulfamoyl)-N-[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl]-N-methylpropanamide

[0582] ##STR00091##

[0583] Analogously to the preparation of Example 29, 185 mg (0.37 mmol) of N-[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl]-N-methyl-3-sulfamoylpropanamide in dichloromethane (9.25 ml) were treated with 37 μl (0.65 mmol) of acetic acid, 125 mg (0.65 mmol) of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (CAS No.: 25952-53-8) and 79.5 mg (0.65 mmol) of 4-dimethylaminopyridine. The mixture was stirred for 18 hours. The reaction was repeated analogously using 20 mg N-[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl]-N-methyl-3-sulfamoylpropanamide. The combined reaction mixtures were diluted with dichloromethane and washed with brine, organic phase was dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by preparative HPLC (Method E). The yield was 140 mg of the title compound.

[0584] LC-MS (method B): R.sub.t=5.12 min; m/z=540.2 (M+H)+

[0585] 1H NMR (400 MHz, DMSO-d6): δ [ppm]=1.02 (s, 3H), 1.38-1.56 (m, 1H), 1.56-1.76 (m, 4H), 1.91 (s, 4H), 2.08-2.20 (m, 2H), 2.27-2.37 (m, 2H), 2.38-2.45 (m, 3H), 2.83-2.91 (m, 2H), 3.10 (s, 3H), 3.50 (dd, 2H), 6.26-6.28 (m, 1H), 7.04-7.12 (m, 2H), 7.34 (d, 1H), 7.66-7.71 (m, 1H), 8.44 (d, 1H), 8.49-8.51 (m, 1H), 11.51 (br. s, 1H).

Example 32

N-[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl]-3-(2H-tetrazol-5-yl)propanamide

[0586] ##STR00092##

[0587] Analogously to the preparation of Intermediate 6-A, 85 mg (0.24 mmol) of 17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-amine (Intermediate 10-A) were treated with 185 mg (0.48 mmol) of 1-[bis-(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxid hexafluorophosphate, 69.3 mg (0.48 mmol) of 3-(2H-tetrazol-5-yl)propanoic acid (CAS No.: 100508-42-7, Afferchem Inc.) and 107 μl (0.97 mmol) of 4-methylmorpholine. The residue was purified by chromatography using a prepacked C18 cartridge (mobile phase: acetonitrile:water 0.1% formic acid buffer, gradient 10% to 90%). The yield was 54 mg (46% of theory) of the title compound.

[0588] LC-MS (method B): R.sub.t=4.81 min; m/z=473.2 (M+H).sup.+

[0589] .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm]=0.99 (s, 3H), 1.36-1.62 (m, 4H), 1.67-1.77 (m, 1H), 1.84-1.92 (m, 1H), 2.08-2.39 (m, 6H), 2.72-2.84 (m, 4H), 3.04-3.15 (m, 2H), 6.25-6.27 (m, 1H), 7.15 (d, 1H), 7.24-7.31 (m, 2H), 7.65-7.70 (m, 1H), 8.43 (d, 1H), 8.48-8.50 (m, 1H), 9.87 (s, 1H).

Example 33

N-[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl]-N-methyl-3-(2H-tetrazol-5-yl)propanamide

[0590] ##STR00093##

[0591] Analogously to the preparation of Intermediate 6-A, 90 mg (0.24 mmol) of 17-(5-fluoropyridin-3-yl)-N-methylestra-1(10),2,4,16-tetraen-3-amine (Intermediate 11-A) were treated with 123 mg (0.32 mmol) of 1-[bis-(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxid hexafluorophosphate, 46.0 mg (0.32 mmol) of 3-(2H-tetrazol-5-yl)propanoic acid (CAS No.: 100508-42-7, Afferchem Inc.) and 71 μl (0.64 mmol) of 4-methylmorpholine. The residue was purified by preparative HPLC (Method E). The yield was 35 mg (24% of theory) of the title compound.

[0592] LC-MS (method B): R.sub.t=5.05 min, m/z=487.1 (M+H).sup.+

[0593] .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm]=1.01 (s, 3H), 1.38-1.52 (m, 1H), 1.56-1.76 (m, 4H), 1.86-1.94 (m, 1H), 2.08-2.19 (m, 2H), 2.26-2.36 (m, 2H), 2.39-2.44 (m, 1H), 2.49-2.56 (m, 2H), 2.82-2.87 (m, 2H), 3.00 (t, 2H), 3.08 (s, 3H), 6.26-6.28 (m, 1H), 6.98-7.08 (m, 2H), 7.33 (d, 1H), 7.66-7.71 (m, 1H), 8.44 (d, 1H), 8.50 (t, 1H).

Examples Type IC4

Example 34

tert-butyl 4-{[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl](methyl)amino}-4-oxobutanoate

[0594] ##STR00094##

[0595] To a solution of 131 mg (0.26 mmol) of tert-butyl 4-{[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl]amino}-4-oxobutanoate (Example 22) in 800 μl of dry N,N-dimethylformamide at 0° C. under argon 81 μl (1.29 mmol) of iodomethane followed by 11.5 mg (0.28 mmol) of sodium hydride (60% in oil) were added. The mixture was stirred for 1 hour at 0° C. and then allowed to warm to RT for 30 minutes. Water was carefully added to the reaction mixture and then extraction into ethyl acetate. The organic phase was further washed with brine, dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by chromatography using a prepacked silica gel cartridge (mobile phase: dichloromethane/ethyl acetate, gradient 0% to 15%). The yield was 32 mg (24% of theory) of the title compound.

[0596] LC-MS (method D): R.sub.t=4.91 min, m/z=519.5 (M+H).sup.+

[0597] .sup.1H-NMR (400 MHz, CDCl.sub.3): δ [ppm]=1.08 (s, 3H), 1.43 (s, 9H), 1.46-1.58 (m, 1H), 1.66-1.76 (m, 3H), 1.76-1.88 (m, 1H), 1.96-2.04 (m, 1H), 2.13-2.24 (m, 2H), 2.31-2.45 (m, 5H), 2.51 (dd, 2H), 2.90-2.97 (m, 2H), 3.24 (s, 3H), 6.11 (dd, 1H), 6.94-6.95 (m, 1H), 6.98 (dd, 1H), 7.31 (d, 1H), 7.38-7.43 (m, 1H), 8.35 (d, 1H), 8.48 (dd, 1H).

Example 35

4-{[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl](methyl)amino}-4-oxobutanoic acid

[0598] ##STR00095##

[0599] Analogously to the preparation of Example 23, 32 mg (0.06 mmol) of tert-butyl 4-{[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl](methyl)amino}-4-oxobutanoate were reacted with 238 μl (3.08 mmol) of trifluoroacetic acid. The crude residue was purified by chromatography using a prepacked silica gel cartridge (mobile phase: ethyl acetate/methanol: 30% triethylamine in water 7:3, gradient 0% to 20%). The yield was 31 mg (83% of theory) of the title compound as a 1.0 equivalent trifluoroacetic acid salt.

[0600] LC-MS (method B): R.sub.t=5.19 min; m/z=463.2 (M+H).sup.+

[0601] .sup.1H-NMR (400 MHz, CD.sub.3OD): δ [ppm]=1.09 (s, 3H), 1.27 (s, 1H), 1.46-1.59 (m, 1H), 1.66-1.75 (m, 3H), 1.82-1.91 (m, 1H), 1.99-2.07 (m, 1H), 2.16-2.25 (m, 2H), 2.31-2.52 (m, 6H), 2.91-2.99 (m, 2H), 3.20 (s, 3H), 6.21 (dd, 1H), 7.01-7.07 (m, 2H), 7.38 (d, 1H), 7.62-7.67 (m, 1H), 8.33 (d, 1H), 8.45 (s, 1H).

Examples Type IC5

Example 36

methyl 3-{[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl](methyl)sulfamoyl}propanoate

[0602] ##STR00096##

[0603] To a solution of 32 mg (0.08 mmol) of 17-(5-fluoropyridin-3-yl)-N-methylestra-1(10),2,4,16-tetraen-3-amine (Intermediate 11-A) in 700 μl of N,N-dimethylformamide under argon at 0° C. 37 μl (0.26 mmol) of triethylamine and 33 mg (0.17 mmol) of 3-chlorosulfonyl-propionic acid methyl ester (CAS No.: 15441-07-3) were added. The mixture was then allowed to warm to RT and stirred for 1 hour. Water was then added and the contents extracted into ethyl acetate. The organic phase was further washed with brine, dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by chromatography using a prepacked silica gel cartridge (mobile phase: cyclohexane/ethyl acetate, gradient 0% to 40%). The yield was 30 mg (60% of theory) of the title compound.

[0604] LC-MS (method A): R.sub.t=4.31 min, m/z=513.4 (M+H).sup.+

[0605] .sup.1H-NMR (300 MHz, CDCl.sub.3): δ [ppm]=1.06 (s, 3H), 1.42-1.57 (m, 3H), 1.61-1.89 (m, 1H), 1.93-2.04 (m, 1H), 2.11-2.24 (m, 2H), 2.29-2.44 (m, 3H), 2.81-2.98 (m, 4H), 3.30-3.33 (m, 6H), 3.72 (d, 3H), 6.10 (dd, 1H), 7.09-7.17 (m, 2H), 7.27-7.32 (m, 1H), 7.36-7.43 (m, 1H), 8.30-8.36 (m, 1H), 8.48 (dd, 1H).

Example 37

methyl 3-{[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl]sulfamoyl}propanoate

[0606] ##STR00097##

[0607] Analogously to the preparation of Example 36, 233 mg (0.67 mmol) of 17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-amine (Intermediate 10-A) were converted into 189 mg (54% of theory) of the title compound.

[0608] LC-MS (method A): R.sub.t=4.16 min; m/z=499.4 (M+H).sup.+

[0609] .sup.1H-NMR (300 MHz, CDCl.sub.3): δ [ppm]=1.06 (s, 3H), 1.40-1.91 (m, 5H), 1.93-2.04 (m, 1H), 2.10-2.24 (m, 2H), 2.27-2.45 (m, 3H), 2.83-2.95 (m, 4H), 3.42 (dd, 2H), 3.71 (d, 3H), 6.10 (dd, 1H), 6.38-6.73 (m, 1H), 7.01 (d, 2H), 7.21-7.26 (m, 1H), 7.37-7.43 (m, 1H), 8.35 (d, 1H), 8.48 (dd, 1H).

Example 38

3-{[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl](methyl)sulfamoyl}propanoic acid

[0610] ##STR00098##

[0611] 30 mg (0.06 mmol) of methyl 3-{[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl](methyl)sulfamoyl}propanoate (Example 36) in 600 μl THF were reacted with 117 μl (0.23 mmol) of 2N lithium hydroxide solution. The mixture was stirred for 16 hours. The reaction mixture was acidified to ˜pH 4 by addition of 1N hydrochloric acid solution and the reaction mixture was concentrated in vacuo. The crude residue was purified by chromatography using a prepacked C18 cartridge (mobile phase: acetonitrile/water 0.1% formic acid buffer, gradient 40% to 98%). The yield was 5.5 mg (18% of theory) of the title compound.

[0612] LC-MS (method B): R.sub.t=5.33 min; m/z=499.2 (M+H).sup.+

[0613] .sup.1H NMR (400 MHz, CD.sub.3OD): δ [ppm]=1.08 (s, 3H), 1.44 (s, 3H), 1.46-1.57 (m, 1H), 1.66-1.74 (m, 3H), 1.80-1.89 (m, 1H), 1.97-2.05 (m, 1H), 2.15-2.24 (m, 2H), 2.32-2.50 (m, 3H), 2.64-2.72 (m, 2H), 2.90-2.97 (m, 2H), 3.35 (t, 2H), 6.21 (dd, 1H), 7.12-7.19 (m, 2H), 7.31 (d, 1H), 7.63 (d, 1H), 8.33-8.34 (m, 1H), 8.43-8.46 (m, 1H)

Example 39

3-{[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl]sulfamoyl}propanoic acid

[0614] ##STR00099##

[0615] Analogously to the preparation of Example 38, 189 mg (0.38 mmol) of methyl 3-{[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl]sulfamoyl}propanoate (Example 37) were reacted with 379 μl (0.75 mmol) of 2N lithium hydroxide solution. The crude residue was purified by preparative HPLC (Method E) to obtain 126 mg (68% of theory) of the title compound.

[0616] LC-MS (method B): R.sub.t=4.95 min; m/z=485.1 (M+H).sup.+

[0617] .sup.1H-NMR (400 MHz, DMSO-d6): δ [ppm]=1.00 (s, 3H), 1.37-1.77 (m, 6H), 1.84-1.92 (m, 1H), 2.07-2.17 (m, 1H), 2.20-2.39 (m, 3H), 2.61 (dd, 2H), 2.77-2.85 (m, 2H), 3.23 (dd, 2H), 6.27 (dd, 1H), 6.89-6.97 (m, 2H), 7.20 (d, 1H), 7.66-7.70 (m, 1H), 8.43-8.50 (m, 2H), 9.65 (s, 1H), 12.52 (br. s, 1H).

Examples Type IC6

Example 40

N-[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl]acetamide

[0618] ##STR00100##

[0619] To a solution of 100 mg (0.28 mmol) of 17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-amine

[0620] (Intermediate 10-A) in 1.5 ml of N,N-dimethylformamide under argon at 0° C. 80 μl (0.57 mmol) of triethylamine and 21 μl (0.30 mmol) of acetyl chloride were added. The mixture was then allowed to warm to RT and stirred for 1 hour. Water and ethyl acetate were added to the reaction mixture and the organic phase was separated and further washed with brine, dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by preparative HPLC (Method E). The yield was 52 mg (46% of theory) of the title compound.

[0621] LC-MS (method B): R.sub.t=5.18 min; m/z=391.2 (M+H).sup.+

[0622] .sup.1H-NMR (400 MHz, DMSO-d.sub.6): δ [ppm]=1.00 (s, 3H), 1.36-1.63 (m, 3H), 1.67-1.77 (m, 1H), 1.84-1.92 (m, 1H), 1.98 (s, 3H), 2.07-2.39 (m, 6H), 2.75-2.84 (m, 2H), 6.27 (dd, 1H), 7.15 (d, 1H), 7.24-7.30 (m, 2H), 7.66-7.70 (m, 1H), 8.43-8.50 (m, 2H), 9.72 (s, 1H).

Examples Type IC7

Example 41

ethyl N-{[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl]carbamoyl}-beta-alaninate

[0623] ##STR00101##

[0624] To a solution of 50 mg (0.14 mmol) of 17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-amine (Intermediate 10-A) in 2.5 ml of dichloromethane at 0° C. under argon 37 mg (0.25 mmol) of ethyl N-(oxomethylidene)-beta-alaninate (CAS No.: 5100-34-5) was added. The reaction mixture was allowed to warm to RT and stirred for 4 hours. It was then concentrated under reduced pressure and the residue was purified using a prepacked silica gel cartridge (mobile phase: cyclohexane/ethyl acetate, gradient 0% to 60%). The yield was 71 mg (96% of theory) of the title compound.

[0625] LC-MS (method A): R.sub.t=4.10 min; m/z=492.4 (M+H).sup.+

[0626] .sup.1H-NMR (300 MHz, CDCl.sub.3): δ [ppm]=1.05 (s, 3H), 1.22-1.28 (m, 3H), 1.42-1.51 (m, 1H), 1.60-1.89 (m, 5H), 1.91-2.02 (m, 1H), 2.09-2.23 (m, 2H), 2.26-2.45 (m, 3H), 2.55-2.60 (m, 2H), 2.86-2.95 (m, 2H), 3.53 (q, 2H), 4.10-4.18 (m, 1H), 5.41 (t, 1H), 6.09 (dd, 1H), 6.45 (s, 1H), 6.97-7.07 (m, 2H), 7.22 (d, 1H), 7.40 (ddt, 1H), 8.34 (d, 1H), 8.48 (t, 1H).

Example 42

N-{[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl]carbamoyl}-beta-alanine

[0627] ##STR00102##

[0628] To a solution of 71 mg (0.13 mmol) of ethyl N-{[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl]carbamoyl}-beta-alaninate (Example 41) in 780 μl of tetrahydrofuran under argon 137 μl (0.27 mmol) of 2N lithium hydroxide solution was added. The mixture was stirred at RT for 16 hours. The reaction mixture was acidified to pH ˜4 by addition of 1N hydrochloric acid solution and the reaction mixture was concentrated in vacuo. The crude residue was purified by preparative HPLC (Method E). The yield was 45 mg (70% of theory) of the title compound.

[0629] LC-MS (method B): R.sub.t=4.79 min; m/z=464.2 (M+H).sup.+

[0630] .sup.1H-NMR (400 MHz, DMSO-d.sub.6): δ [ppm]=1.00 (s, 3H), 1.35-1.61 (m, 4H), 1.67-1.76 (m, 1H), 1.83-1.91 (m, 1H), 2.10-2.39 (m, 9H), 2.73-2.82 (m, 2H), 6.12 (dd, 1H), 6.26 (dd, 1H), 7.08 (d, 3H), 7.65-7.70 (m, 1H), 8.35 (s, 1H), 8.42-8.50 (m, 2H), 12.2 (br. s, 1H).

Examples Type IC8

Example 43

methyl 4-{[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl](methyl)amino}butanoate

[0631] ##STR00103##

[0632] To a solution of 150 mg (0.41 mmol) of 17-(5-fluoropyridin-3-yl)-N-methylestra-1(10),2,4,16-tetraen-3-amine (Intermediate 11-A) in 600 μl of acetonitrile and 300 μl of N,N-dimethylformamide under argon 86 mg (0.62 mmol) of potassium carbonate followed by 82 mg (0.45 mmol) of methyl 4-bromobutanoate (CAS No.: 4897-84-1) were added. The reaction mixture was stirred at 65° C. for 16 hours, allowed to cool to RT and partitioned between water and ethyl acetate. The phases were separated, the organic phase was further washed with water, brine, dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by chromatography using a prepacked silica gel cartridge (mobile phase: dichloromethane/ethyl acetate, gradient 0% to 70%). The yield was 163 mg (81% of theory) of the title compound.

[0633] LC-MS (method C): R.sub.t=3.76 min; m/z=463.1 (M+H).sup.+

[0634] .sup.1H NMR (300 MHz, CDCl.sub.3): δ [ppm]=1.04 (s, 3H), 1.45-1.98 (m, 7H), 2.09-2.20 (m, 3H), 2.32-2.43 (m, 6H), 2.81-2.96 (m, 4H), 3.32 (t, 2H), 3.68 (s, 3H), 6.07-6.11 (m, 1H), 6.45-6.48 (m, 1H), 6.57 (dd, 1H), 7.15 (d, 1H), 7.37-7.43 (m, 1H), 8.34 (d, 1H), 8.48 (s, 1H).

Example 44

4-{[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl](methyl)amino}butanoic acid

[0635] ##STR00104##

[0636] Analogously to the preparation of Example 42, 160 mg (0.34 mmol) of methyl 4-{[17-(5-fluoropyridin-3-yl)estra-1(10),2,4,16-tetraen-3-yl](methyl)amino}butanoate (Example 43) were reacted with 346 μl (0.69 mmol) of 2N lithium hydroxide solution. The crude residue was purified by preparative HPLC (Method E) to obtain 76 mg (48.5% of theory) of the title compound.

[0637] LC-MS (method B): R.sub.t=4.34 min; m/z=449.2 (M+H).sup.+

[0638] .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm]=0.99 (s, 3H), 1.33-1.76 (m, 6H), 1.82-1.90 (m, 1H), 2.07-2.22 (m, 4H), 2.26-2.36 (m, 2H), 2.77-2.79 (m, 4H), 3.15-3.28 (m, 5H), 6.25-6.27 (m, 1H), 6.38 (d, 1H), 6.49 (dd, 1H), 7.02 (d, 1H), 7.65-7.70 (m, 1H), 8.43 (d, 1H), 8.49-8.50 (m, 1H).

Examples Type IC9

Example 45

4-{[17-(6-methylpyridazin-4-yl)estra-1(10),2,4,16-tetraen-3-yl]amino}-4-oxobutanoic acid

[0639] ##STR00105##

[0640] Analogously to the preparation of Example 24, 73 mg (0.14 mmol) of methyl 4-oxo-4-{[17-{[(trifluoromethyl)sulfonyl]oxy}estra-1(10),2,4,16-tetraen-3-yl]amino}butanoate (Intermediate 12-A) were reacted with 3-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridazine (CAS No.: 1350543-95-1) and 7.95 mg (0.01 mmol) of dichlorobis(triphenylphosphine)palladium(II). After 4 hours at 95° C., the reaction mixture was allowed to cool to 50° C. and 71 μl of 2N lithium hydroxide solution were added. After 2 hours of stirring the reaction mixture was allowed to cool to room temperature and concentrated in vacuo. The residue was partially dissolved in a solution of 5% of methanol in dichloromethane, filtered and purified by chromatography using a prepacked silica gel cartridge (mobile phase: dichloromethane/dichloromethane:methanol:acetic acid:water 240:20:3:2, gradient 0% to 50%). The yield was 21 mg (33% of theory) of the title compound.

[0641] LC-MS (method B): R.sub.t=3.57 min, m/z=446.2 (M+H).sup.+

[0642] .sup.1H NMR (400 MHz, DMSO-d6): δ [ppm]=1.02 (s, 3H), 1.36-1.72 (m, 6H), 1.83-1.92 (m, 1H), 2.09-2.40 (m, 6H), 2.43-2.51 (m, 1H), 2.59 (s, 3H), 2.63-2.84 (m, 3H), 6.56 (dd, 1H), 7.15 (d, 1H), 7.24-7.31 (m, 2H), 7.49 (d, 1H), 9.11 (d, 1H), 9.79 (s, 1H), 12.03 (br. s, 1H).

Examples Type IC10

Example 46

17-(5-fluoropyridin-3-yl)-3-(S-methylsulfonimidoyl)estra-1(10),2,4,16-tetraene

[0643] ##STR00106##

[0644] Analogously to the preparation of Example 24, 106 mg (0.161 mmol) of 3-[S-methyl-N-(trifluoroacetyl)sulfonimidoyl]estra-1(10),2,4,16-tetraen-17-yl trifluoromethanesulfonate (Intermediate 15-A) were treated with 31.7 mg (0.22 mmol) of 5-fluoropyridine-3-boronic acid (CAS No.: 872041-86-6) and 9.0 mg (0.013 mmol) of dichlorobis(triphenylphosphine)palladium(II) (CAS No.: 13965-03-2). The residue was purified using a prepacked silica gel cartridge (mobile phase: dichloromethane/ethyl acetate, gradient 0% to 80%) and then by chromatography using a prepacked C18 cartridge (mobile phase: acetonitrile/water 0.1% formic, gradient 10% to 90%). The yield was 22.2 mg (33% of theory) of the title compound.

[0645] LC-MS (method B): R.sub.t=4.59 min; m/z=411.1 (M+H).sup.+

[0646] .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ [ppm]=1.01 (s, 3H), 1.45-1.77 (m, 6H), 1.89-1.96 (m, 1H), 2.09-2.20 (m, 2H), 2.29-2.43 (m, 3H), 2.94 (dd, 2H), 3.00 (s, 3H), 4.02 (s, 1H), 6.28 (dd, 1H), 7.47 (d, 1H), 7.60-7.66 (m, 2H), 7.67-7.72 (m, 1H), 8.49-8.51 (m, 1H).

[0647] Biological Assays:

[0648] Example compounds were tested in selected biological assays one or more times. When tested more than once, data are reported as either average values or as median values, wherein [0649] the average value, also referred to as the arithmetic mean value, represents the sum of the values obtained divided by the number of times tested, and [0650] the median value represents the middle number of the group of values when ranked in ascending or descending order. If the number of values in the data set is odd, the median is the middle value. If the number of values in the data set is even, the median is the arithmetic mean of the two middle values.

[0651] Example compounds were synthesized one or more times. When synthesized more than once, data from biological assays represent average values or median values calculated utilizing data sets obtained from testing of one or more synthetic batch.

Example 47 (AKR1C3-Inhibitory Action)

[0652] The AKR1C3-inhibitory action of the substances of this invention was measured in the AKR1C3 assay described in the paragraphs which follow.

[0653] Essentially, the enzyme activity is measured by quantifying the coumberol formed from coumberone (Halim, M., Yee, D., J., and Sames, D., J. A., M. (2008). Chem Soc 130, 14123-14128 and Yee, D., J., Balsanek, V., Bauman, D., R., Penning, T., M., and Sames, D. (2006). Proc Natl Acad Sci USA, 103, 13304-13309). In this assay, the increase in the highly fluorescent coumberol can be determined by NADPH (nicotinamide adenine dinucleotide phosphate)-dependent reduction of the nonfluorescent coumberone by AKR1C3.

[0654] The enzyme used was recombinant human AKR1C3 (aldo-keto reductase family 1 member C.sub.3) (GenBank Accession No. NM_003739). This was expressed as the GST (glutathione S-transferase) fusion protein in E. coli and purified by means of glutathione-Sepharose affinity chromatography. The GST was removed by thrombin digestion with subsequent size exclusion chromatography (Dufort, I., Rheault, P., Huang, X., F., Soucy, P., and Luu-The, V. (1999). Endocrinology,140, 568-574).

[0655] For the assay, 40-50 nl of a 100-fold concentrated solution of the test substance in DMSO were pipetted into a black low-volume 384-well microtitre plate (Greiner Bio-One, Frickenhausen, Germany), 2-2.5 μl of a solution of AKR1C3 in assay buffer [50 mM potassium phosphate buffer pH 7, 1 mM DTT, 0.0022% (w/v) Pluronic F-127, 0.01% BSA (w/v) and protease inhibitor cocktail (complete, EDTA-free Protease Inhibitor Cocktail from Roche)] were added and the mixture was incubated for 20 min, in order to enable preliminary binding of the substances to the enzyme prior to the enzyme reaction. Then the enzyme reaction was started by adding 2-2.5 μl of a solution of NADPH and coumberone in assay buffer (final assay concentration was 10 μM NADPH and 0.3 μM coumberone) and the resulting mixture was incubated at room temperature for 3 h. Typical concentrations were depending on the linear range of the enzyme reaction during the reaction time about 1 nM. The reaction was stopped by adding 2 μl of a stop solution consisting of the AKR1C3 inhibitor EM-1404 to a final concentration of 1 μM. Subsequently, the fluorescence of coumberol was measured at 520 nm (excitation at 380 nm) with a suitable measuring instrument (Pherastar, BMG). The intensity of the fluorescence was used as a parameter for the amount of coumberol formed and hence for the enzyme activity of AKR1C3. The data were normalized (enzyme+coumberon substrate in assay buffer=0% inhibition; coumberone substrate only in assay buffer=100% inhibition). Substances were usually tested at 11 different concentrations on the same microtiter plate in a range between 96.8 pM to 20 μM (20 μM, 5.9 μM, 1.7 μM, 0.5 μM, 0.15 μM, 44 nM, 12.9 nM, 3.8 nM, 1.1 nM, 0.3 nM and 96.8 pM). The dilutions were carried out on a 100 fold concentrated solution by serial dilution in duplicates. From the obtained data IC.sub.50 values were calculated by a 4-parameter fit.

[0656] As described, the pharmacological substances claimed were tested for their inhibitory effect on the AKR1C3 enzyme (see Table 1). The compounds tested show strong inhibition of AKR1C3 in vitro (IC.sub.50 values<500 nM) and predominantly even IC.sub.50 values<20 nM.

TABLE-US-00002 TABLE 1 Inhibition of AKR1C3 by the compounds of the present invention (mean values) Example compound AKR1C3 enzyme inhibition IC.sub.50 [nmol/l] 01 02 3.5 03 2.4 04 05 91.7 06 07 0.5 08 4.1 09 7.7 10 2.9 11 12 12.0 13 14 14.3 15 73.0 16 9.7 17 1.6 18 33.5 19 1.7 20 3.2 21 5.3 22 23 3.8 24 25 313 26 8.6 27 5.0 28 29 3.2 30 31 4.1 32 11.8 33 2.5 34 35 3.2 36 37 38 4.9 39 3.0 40 5.3 41 42 8.3 43 44 18.5 45 1.8 46 2.2

Example 48 (Test of AKR1C3 Inhibition in a Cell-Based System)

[0657] The inhibition of AKR1C3 by the substances described in this invention was measured in a cell-based assay using coumberon as the substrate for AKR1C3 (Halim, M., Yee, D., J., and Sames, D., J. (2008). Am Chem Soc, 130, 14123-14128 and Yee, D., J., Balsanek, V., Bauman, D., R., Penning, T., M., and Sames, D. (2006), Proc Natl Acad Sci USA 103, 13304-13309) (cf. Example 48).

[0658] The cell system used was HEK293 cells (ATCC, USA) (DMEM/F12, 10% FCS, 2 mM L-Glutamine, PSG). The cells were transfected with an AKR1C3 expression plasmid (pCMV6-AC-AKR1C3, GenBank Accession No. NM 003739.4), harvested after 24 hours and subsequently frozen. For assays, the frozen cells were thawed in assay medium (DMEM/F12 w/o phenolred, 0.005% Tween, 10% FCS, 2 mM L-Glutamine, PSG). Cells were seeded at 2500 cells/well into 384-well low volume or 1536-well plates (Greiner Bio-One, Frickenhausen, Germany) that contained 40-50 nl of a 100 fold concentrated compound solution in DMSO. After incubation at room temperature for 15 minutes, coumberon in PBS was added to a final concentration of 6×10.sup.−6M. Following a 3 h incubation at room temperature, the reaction was stopped by lysing the cells with 0.4% Triton X-100. Coumberol fluorescence intensity was measured at 520 nm (excitation 380 nm) with a suitable measuring instrument (Pherastar, BMG). The intensity of the fluorescence was used as a parameter for the amount of coumberol formed and hence for the enzyme activity of AKR1C3. The data were normalized (transfected cells+coumberon=0% inhibition; no cells+coumberone=100% inhibition). Substances were usually tested at 11 different concentrations on the same microtiter plate in a range between 96.8 pM to 20 μM (20 μM, 5.9 μM, 1.7 μM, 0.5 μM, 0.15 μM, 44 nM, 12.9 nM, 3.8 nM, 1.1 nM, 0.3 nM and 96.8 pM). The dilutions were carried out on a 100 fold concentrated solution by serial dilution in duplicates. From the obtained data IC.sub.50 values were calculated by a 4-parameter fit.

[0659] The pharmacological substances claimed were tested for their inhibitory action on the AKR1C3 enzyme by means of the cell-based assay described above (see Table 2). The compounds tested exhibited a significant inhibition of cellular AKR1C3 in vitro, most of them show IC50<1 μM.

TABLE-US-00003 TABLE 2 Inhibition of AKR1C3 by the compounds of the present invention in a cellular assay (mean values) Cellular AKR1C3 Example compound inhibition IC.sub.50 [μmol/l] 01 02 0.38 03 0.05 04 05 2.71 06 07 0.80 08 0.08 09 0.05 10 0.49 11 12 0.70 13 14 0.49 15 3.18 16 0.54 17 0.17 18 1.79 19 0.15 20 0.03 21 0.04 22 23 0.78 24 25 8.34 26 0.15 27 0.11 28 29 5.00 30 31 1.54 32 2.17 33 0.26 34 35 0.21 36 37 38 0.86 39 5.10 40 0.10 41 42 8.09 43 44 1.12 45 0.98 46 0.03

Example 49: Determination of Antiandrogenic Action

[0660] The antiandrogenic action of the substance was measured in adult monkeys (Macaca fascicularis), as a surrogate for the antiproliferative effects in prostate cancer and metastases thereof. The monkeys (4 per group) were treated by the oral route by means of a gavage with 1, 3 or 10 mg/kg substance or with vehicle over 4 weeks. The size of the prostate and of the seminal vesicle was determined by ultrasound at the start of the experiment and after one, two, three and four weeks. The decrease in the weight of these organs was taken as evidence for the antiandrogenicity of the substances. In addition, the blood concentrations (in the serum or in the plasma) of various steroids (DHEA, testosterone, androstenedione, hydroxyprogesterone) and prostaglandins (PGD2, PGD2, PGF2alpha) were determined at the start of the experiment and after one, two, three or four weeks. Since AKR1C3 is involved both in the steroid synthesis route and in the prostaglandin synthesis route, changes in the blood concentrations of these steroids and prostaglandins are taken as an indication of the in vivo effect of the substance.