Inhibitors of glucocorticoid receptor

Abstract

The present invention relates generally to compositions and methods for treating cancer and hypercortisolism. Provided herein are substituted steroidal derivative compounds and pharmaceutical compositions comprising said compounds. The subject compounds and compositions are useful for inhibition of glucocorticoid receptors. Furthermore, the subject compounds and compositions are useful for the treatment of cancer and hypercortisolism.

Claims

1. A compound having the structure of Formula (I), or a pharmaceutically acceptable salt thereof: ##STR00257## wherein R.sup.1 is —NR.sup.3S(O).sub.2R.sup.6, —NR.sup.3S(O).sub.2NR.sup.3R.sup.4, -alkylNR.sup.3S(O).sub.2R.sup.6, -alkylNR.sup.3S(O).sub.2NR.sup.3R.sup.4, —NR.sup.3S(O)R.sup.6, —Oaralkyl, —C(O)NR.sup.3R.sup.4, —OC(O)OR.sup.5, —C(O)OR.sup.5, —OC(O)NR.sup.3R.sup.4, —S(O).sub.2NR.sup.3R.sup.4, —S(O).sub.2R.sup.6, or —S(O)R.sup.6; R.sup.3 and R.sup.4 are each independently —H, optionally substituted alkyl, haloalkyl, optionally substituted carbocyclyl, optionally substituted carbocyclylalkyl, optionally substituted heteroalkyl, optionally substituted aryl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl, optionally substituted heteroaryl, —S(O).sub.2R.sup.6, —C(O)N(R.sup.10).sub.2, —C(O)R.sup.5, or —C(O)OR.sup.5; or R.sup.3 and R.sup.4 attached to the same N atom are taken together with the N atom to which they are attached to form a substituted or unsubstituted heterocycle; R.sup.5 is optionally substituted alkyl, haloalkyl, optionally substituted carbocyclyl, optionally substituted aryl, optionally substituted heterocyclyl, or optionally substituted heteroaryl; R.sup.6 is optionally substituted alkyl, haloalkyl, optionally substituted carbocyclyl, optionally substituted heteroalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heterocyclyl, or optionally substituted heteroaryl; R.sup.7 and R.sup.8 are each independently —H, optionally substituted alkyl, haloalkyl, halo, optionally substituted carbocyclyl, optionally substituted carbocyclylalkyl, optionally substituted heteroalkyl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl, —OH, —OR.sup.5, —NR.sup.3R.sup.4, —C(O)NR.sup.3R.sup.4, —CN, —S(O).sub.2R.sup.6, —C(O).sub.2H, —C(O)R.sup.5, or —C(O)OR.sup.5; or R.sup.7 and R.sup.8 are taken together with the atom to which they are attached to form a substituted or unsubstituted ring containing 0-2 heteroatoms selected from the group consisting of —O—, —NH—, —NR.sup.6—, —S—, and —S(O).sub.2; and each R.sup.10 is independently H, optionally substituted alkyl, haloalkyl, optionally substituted carbocyclyl, optionally substituted heteroalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heterocyclyl, or optionally substituted heteroaryl.

2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein R.sup.1 is —NR.sup.3S(O).sub.2R.sup.6, —NR.sup.3S(O).sub.2NR.sup.3R.sup.4, -alkylNR.sup.3S(O).sub.2R.sup.6, -alkylNR.sup.3S(O).sub.2NR.sup.3R.sup.4, —C(O)NR.sup.3R.sup.4, —S(O).sub.2NR.sup.3R.sup.4, —S(O).sub.2R.sup.6; R.sup.3 and R.sup.4 are each independently —H, optionally substituted alkyl, fluoroalkyl, optionally substituted carbocyclyl, optionally substituted carbocyclylalkyl, optionally substituted aryl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl, optionally substituted heteroaryl, —C(O)N(R.sup.10).sub.2, or —S(O).sub.2R.sup.6; or R.sup.3 and R.sup.4 attached to the same N atom are taken together with the N atom to which they are attached to form a substituted or unsubstituted 3-, 4-, 5-, or 6-membered heterocycle; R.sup.5 is optionally substituted alkyl, fluoroalkyl, optionally substituted carbocyclyl, or optionally substituted heterocyclyl; R.sup.6 is optionally substituted alkyl, fluoroalkyl, optionally substituted carbocyclyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heterocyclyl, or optionally substituted heteroaryl; R.sup.7 and R.sup.8 are each independently —H, optionally substituted alkyl, fluoroalkyl, halo, —OR.sup.5, —NR.sup.3R.sup.4, —C(O)NR.sup.3R.sup.4, —CN, optionally substituted carbocyclyl, or optionally substituted carbocyclylalkyl; or R.sup.7 and R.sup.8 are taken together with the atom to which they are attached to form a substituted or unsubstituted 3-, 4-, 5-, or 6-membered ring containing 0-2 heteroatoms selected from the group consisting of —O—, —NH—, —NR.sup.6—, —S—, and —S(O).sub.2; and each R.sup.10 is independently H, optionally substituted alkyl, haloalkyl, optionally substituted carbocyclyl, optionally substituted heteroalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heterocyclyl, or optionally substituted heteroaryl.

3. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein R.sup.1 is —NR.sup.3S(O).sub.2R.sup.6, —C(O)NR.sup.3R.sup.4, —S(O).sub.2NR.sup.3R.sup.4, —S(O).sub.2R.sup.6.

4. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein R.sup.1 is —NR.sup.3S(O).sub.2R.sup.6 or —S(O).sub.2NR.sup.3R.sup.4.

5. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein R.sup.3 and R.sup.4 are each independently —H, optionally substituted alkyl, fluoroalkyl, optionally substituted carbocyclyl, optionally substituted carbocyclylalkyl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl,—C(O)N(R.sup.10).sub.2, or —S(O).sub.2R.sup.6.

6. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein R.sup.3 and R.sup.4 are each independently —H, alkyl, or —S(O).sub.2R.sup.6.

7. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein R.sup.3 and R.sup.4 attached to the same N atom are taken together with the N atom to which they are attached to form a substituted or unsubstituted 4-, 5-, or 6-membered ring heterocycle containing 0-3 heteroatoms selected from —O—, —NH—, —NR.sup.5—, —S—, and —S(O).sub.2—; and R.sup.5 is alkyl.

8. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein R.sup.6 is alkyl, carbocyclyl, optionally substituted aryl, optionally substituted aralkyl, or optionally substituted heterocyclyl.

9. The compound of claim 8, or a pharmaceutically acceptable salt thereof, wherein R.sup.6 is C.sub.1-6 alkyl, C.sub.3-6 carbocyclyl, optionally substituted phenyl, optionally substituted benzyl, or optionally substituted heterocyclyl.

10. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein R.sup.7 and R.sup.8 are each independently —H, alkyl, or carbocyclyl.

11. The compound of claim 10, or a pharmaceutically acceptable salt thereof, wherein R.sup.7 and R.sup.8 are —H.

12. A pharmaceutical composition comprising the compound of claim 1, or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable excipient.

Description

EXAMPLES

(1) I. Chemical Synthesis

(2) Unless otherwise noted, reagents and solvents were used as received from commercial suppliers. Anhydrous solvents and oven-dried glassware were used for synthetic transformations sensitive to moisture and/or oxygen. Yields were not optimized. Reaction times are approximate and were not optimized. Column chromatography and thin layer chromatography (TLC) were performed on silica gel unless otherwise noted.

(3) The substituted steroidal derivative compounds are prepared by the general synthetic route described in Scheme 1.

Example 1: Synthesis of (8S,11R,13S,14S,17S)-11-(3,5-Difluoro-4-methylphenyl)-17-hydroxy-13-methyl-17-(prop-1-yn-1-yl)-1,2,6,7,8,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenanthren-3-one

(4) ##STR00171##

Step A. (8S,13S,14S,17S)-13-Methyl-17-(prop-1-yn-1-yl)-1,2,4,6,7,8,12,13,14,15,16,17-dodecahydrospiro[cyclopenta[a]phenanthrene-3,2′-[1,3]dioxolan]-17-ol

(5) ##STR00172##

(6) To a stirred solution of (8S,13S,14S)-13-methyl-1,4,6,7,8,12,13,14,15,16-decahydrospiro[cyclopenta[a]phenanthrene-3,2′-[1,3]dioxolan]-17(2H)-one (3.4 g, 10.9 mmol) in THF (45 mL) at 0° C. was added bromo(prop-1-ynyl)magnesium (0.5 M in THF, 65.4 mL, 32.7 mmol) slowly. The reaction was allowed to warm up to rt and stirred for 60 min. The reaction mixture was quenched (sat. NH.sub.4Cl), extracted (EtOAc), and washed (brine). The combined organic layers were dried (Na.sub.2SO.sub.4) and concentrated under reduced pressure. The residue was purified by flash column chromatography to provide 3.0 g of the title compound as a white solid.

Step B. (5′R,8′S,10′R,13′S,14′S,17′S)-13′-Methyl-17′-(prop-1-yn-1-yl)-1′,2′,7′,8′,12′,13′,14′,15′,16′,17′-decahydro-4′H,6′H-spiro[[1,3]dioxolane-2,3′-[5,10]epoxycyclopenta[a]phenanthren]-17′-ol

(7) ##STR00173##

(8) To a stirred solution of (13S,16R,17S)-13-methyl-17-prop-1-ynyl-spiro[1,2,4,6,7,8,12,14,15,16-decahydrocyclopenta[a]phenanthrene-3,2′-1,3-dioxolane]-17-ol (Step A) (2.88 g, 8.14 mmol) in DCM (60 mL) at 0° C. was added Na.sub.2HPO.sub.4 (2.89 g, 20.35 mmol), followed by hexafluoroacetone trihydrate (1.12 mL, 8.14 mmol) and H.sub.2O.sub.2 (30% aq, 2.0 mL, 24.4 mmol). The mixture was stirred at 0° C. for 10 min and then at rt for 2 h. The resulting mixture was cooled in ice bath and 10% Na.sub.2S.sub.2O.sub.3 aq. solution (8 ml) was added slowly. After being stirred at rt for 30 min, the mixture was quenched (sat. NH.sub.4Cl), extracted (2× EtOAc), and washed (brine). The combined organic layers were dried (Na.sub.2SO.sub.4) and concentrated under reduced pressure. The residue was purified by flash column chromatography to provide 1.8 g of the title compound as a white solid.

Step C. (5R,8S,11R,13S,14S,17S)-11-(3,5-Difluoro-4-methylphenyl)-13-methyl-17-(prop-1-yn-1-yl)-1,2,6,7,8,11,12,13,14,15,16,17-dodecahydrospiro[cyclopenta[a]phenanthrene-3,2′-[1,3]dioxolane]-5,17(4H)-diol

(9) ##STR00174##

(10) A small piece of iodine was added to magnesium pieces (757.88 mg, 31.18 mmol) under argon and the mixture was heated with heat-gun to activate magnesium metal. A solution of 5-bromo-1,3-difluoro-2-methylbenzene (5.28 g, 25.5 mmol) in THF (25 mL) was added to the mixture dropwise. The mixture was stirred at rt for 30 min and then at 55° C. for 1 h. To a solution of copper(I) iodide (539.8 mg, 2.8 mmol) and (5′R,8′S,10′R,13′5,14′S,17′S)-13′-methyl-17′-(prop-1-yn-1-yl)-1′,2′,7′,8′,12′,13′,14′,15′,16′,17′-decahydro-4′H,6′H-spiro[[1,3]dioxolane-2,3′-[5,10]epoxycyclopenta[a]phenanthren]-17′-ol (Step B) (2.1 g, 5.67 mmol) in THF (60 mL) was added the Grignard reagent (26 mL) prepared above dropwise at 0° C. After being stirred at 0° C. for 30 min, the reaction mixture was quenched (sat. NH.sub.4Cl), extracted (EtOAc), and washed (brine). The combined organic layers were dried (Na.sub.2SO.sub.4) and concentrated under reduced pressure. The residue was purified by flash column chromatography provided the title compound as a colorless foam.

Step D. (8S,11R,13S,14S,17S)-11-(3,5-Difluoro-4-methylphenyl)-17-hydroxy-13-methyl-17-(prop-1-yn-1-yl)-1,2,6,7,8,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenanthren-3-one

(11) A solution of (5R,8S,11R,13S,14S,17S)-11-(3,5-Difluoro-4-methylphenyl)-13-methyl-17-(prop-1-yn-1-yl)-1,2,6,7,8,11,12,13,14,15,16,17-dodecahydrospiro[cyclopenta[a]phenanthrene-3,2′-[1,3]dioxolane]-5,17(4H)-diol (Step C) (2.0 g, 4.0 mmol) in 70% acetic acid aq. solution (8.0 mL) was stirred at 55° C. for 1.5 h. The reaction mixture was cooled to rt, quenched (sat. NaHCO.sub.3), extracted (EtOAc), and washed (brine). The combined organic layers were dried (MgSO4), and concentrated under reduced pressure. Purification of the residue by flash column chromatography provided 1.3 g of the title compound as a white solid. .sup.1H NMR (400 MHz, CHLOROFORM-d) δ ppm 6.65-6.73 (2H, m),5.82 (1H, s), 4.37 (1H, brd, J=7.45 Hz),2.69-2.81 (1H, m),2.57-2.65 (2H, m), 2.22-2.52 (6H, m), 2.16 (3H, s), 2.08-1.94 (3H, m), 1.92 (3H, s), 1.66-1.84 (2H, m), 1.32-1.53 (2H, m), 0.55 (3H, s). m/z (ESI, +ve ion) 437.3 (M+H).sup.+.

Example 2: Synthesis of (8S,11R,13S,14S,16R,17S)-11-(4-(Dimethylamino)phenyl)-16-ethyl-17-hydroxy-13-methyl-17-(prop-1-yn-1-yl)-1,2,6,7,8,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenanthren-3-one or (8S,11R,13S,14S,16S,17S)-11-(4-(dimethylamino)phenyl)-16-ethyl-17-hydroxy-13-methyl-17-(prop-1-yn-1-yl)-1,2,6,7,8,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenanthren-3-one

(12) ##STR00175##

Step A. (8S,13S,14S,16R)-16-Ethyl-13-methyl-1,4,6,7,8,12,13,14,15,16-decahydrospiro[cyclopenta[a]phenanthrene-3,2′-[1,3]dioxolan]-17(2H)-one or (8S,13S,14S,16S)-16-ethyl-13-methyl-1,4,6,7,8,12,13,14,15,16-decahydrospiro[cyclopenta[a]phenanthrene-3,2′-[1,3]dioxolan]-17(2H)-one

(13) ##STR00176##

(14) A solution of (8S,13S,14S)-13-methyl-1,4,6,7,8,12,13,14,15,16-decahydrospiro[cyclopenta[a]phenanthrene-3,2′-[1,3]dioxolan]-17(2H)-one (1.00 g, 3.18 mmol) and 0.8 mL of DMPU in THF (3 mL) was added dropwise to a solution [bis(trimethylsilyl)amino]lithium (6.36 mL, 6.36 mmol) in THF (10 mL) at −45° C. After being stirred at −40° C. for 30 min, iodoethane (1.28 mL, 15.9 mmol) was added. The reaction was allowed to warm up to rt and stirred for another 1 h. The reaction was quenched with sat. NH.sub.4Cl, extracted with EtOAc. The organics were washed with water, brine solution, separated, and dried (Na.sub.2SO.sub.4) before concentrating to dryness. The crude was purified by flash column chromatography to give the title compound as a white solid.

Step B. (8S,11R,13S,14S,16R,17S)-11-(4-(Dimethylamino)phenyl)-16-ethyl-17-hydroxy-13-methyl-17-(prop-1-yn-1-yl)-1,2,6,7,8,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenanthren-3-one or (8S,11R,13S,14S,16S,17S)-11-(4-(dimethylamino)phenyl)-16-ethyl-17-hydroxy-13-methyl-17-(prop-1-yn-1-yl)-1,2,6,7,8,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenanthren-3-one

(15) The title compound was prepared from (8S,13S,14S,16R)-16-ethyl-13-methyl-1,4,6,7,8,12,13,14,15,16-decahydrospiro[cyclopenta[a]phenanthrene-3,2′-[1,3]dioxolan]-17(2H)-one or (8S,13S,14S,16S)-16-ethyl-13-methyl-1,4,6,7,8,12,13,14,15,16-decahydrospiro[cyclopenta[a]phenanthrene-3,2′-[1,3]dioxolan]-17(2H)-one (Step A) by procedures similar to those described in Example 1, Steps A-D. .sup.1H NMR (400 MHz, CDCl.sub.3) δ ppm 7.01 (2H, d, J=8.4 Hz), 6.60 (2H, d, J=8.4 Hz), 5.75 (1H, s), 4.36 (1H, d, J=6.2 Hz), 2.90 (6H, s), 2.80-1.91 (8H, m), 1.90 (3H, s), 1.75-1.20 (9H, m), 0.91 (3H, t, J=5.6 Hz), 0.56 (3H, s). m/z (ESI, +ve ion) 458.3 (M+H).sup.+.

Example 3: Synthesis of (8S,11R,13S,14S,17S)-11-(3,5-dichloro-4-(dimethylamino)phenyl)-17-hydroxy-13-methyl-17-(prop-1-yn-1-yl)-1,2,6,7,8,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenanthren-3-one

(16) ##STR00177##

(17) Trichloroisocyanuric acid (63.4 mg, 0.246 mmol) was added to a solution of mifepristone [TCI M1732] (0.157 g, 0.366 mmol) in DCM (1.5 ml). The slurry was allowed to stir at rt for 15 h. Water and more DCM were added and the mixture was washed with sat. NaHCO.sub.3 solution. The organic layer was dried over magnesium sulfate, filtered and concentrated under vacuum. The residue was fractionated by silica gel chromatography. Fractions containing the desired compound were combined and repurified by flash chromatography to afford 33.5 mg (18%) of the title compound. .sup.1H NMR (400 MHz, CDCl3) δ ppm 7.05 (2H, s), 5.81 (1H, s), 4.32 (1H, d, J=7.2 Hz) 2.87 (6H, s), 2.60 (1H, br s) 1.7-2.9 (17H, various m), 1.58 (2H, br s), 0.55 (3H, s). m/z (ESI, +ve ion) 498.3, 500.3 [Cl2 isotope pattern] (M+H).sup.+

Example 4: (8S,11R,13S,14S,17S)-11-(2,3-Dihydrobenzo[b][1,4]dioxin-6-yl)-17-hydroxy-13-methyl-17-(prop-1-yn-1-yl-d3)-1,2,6,7,8,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenanthren-3-one

(18) ##STR00178##

Step A. (8S,13S,14S,17R)-17-Ethynyl-13-methyl-1,2,4,6,7,8,12,13,14,15,16,17-dodecahydrospiro[cyclopenta[a]phenanthrene-3,2′-[1,3]dioxolan]-17-ol

(19) ##STR00179##

(20) To a stirred solution of (8S,13S,14S)-13-methyl-1,4,6,7,8,12,13,14,15,16-decahydrospiro[cyclopenta[a]phenanthrene-3,2′-[1,3]dioxolan]-17(2H)-one (2 g, 6.4 mmol) in THF (12 mL) at 0° C. was added ethynylmagnesium bromide (0.5 M in THF, 44.5 mL, 22.3 mmol) slowly. The reaction was allowed to warm up to rt and stirred for 60 min. The reaction mixture was quenched (sat. NH.sub.4Cl), extracted (EtOAc), and washed (brine). The combined organic layers were dried (Na.sub.2SO.sub.4) and concentrated under reduced pressure. The residue was purified by flash column chromatography to provide 1.1 g of the title compound as a white solid.

Step B. (8S,13S,14S,17S)-13-Methyl-17-(prop-1-yn-1-yl-d3)-1,2,4,6,7,8,12,13,14,15,16,17-dodecahydrospiro[cyclopenta[a]phenanthrene-3,2′-[1,3]dioxolan]-17-ol

(21) ##STR00180##

(22) N-Butyllithium (1.84 mL, 2.94 mmol) was added to a solution of (8S,13S,14S,17R)-17-ethynyl-13-methyl-spiro[1,2,4,6,7,8,12,14,15,16-decahydrocyclopenta[a]phenanthrene-3,2′-1,3-dioxolane]-17-ol (Step A) (500 mg, 1.47 mmol) in THF (2 mL) at 0° C. After being stirred at 0° C. for 10 min, trideuterio(iodo)methane (0.1 mL, 1.62 mmol) was added slowly. The reaction was stirred at rt overnight, then quenched with sat. NH.sub.4Cl, and extracted with EtOAc. The organics were washed with water and brine solution, separated and dried (MgSO.sub.4) before concentration to dryness. The crude material was purified by flash column chromatography to give the title compound as a white solid.

Step C. (8S,11R,13S,14S,17S)-11-(2,3-Dihydrobenzo[b][1,4]dioxin-6-yl)-17-hydroxy-13-methyl-17-(prop-1-yn-1-yl-d3)-1,2,6,7,8,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenanthren-3-one

(23) The title compound was prepared from (8S,13S,14S,17S)-13-methyl-17-(prop-1-yn-1-yl-d3)-1,2,4,6,7,8,12,13,14,15,16,17-dodecahydrospiro[cyclopenta[a]phenanthrene-3,2′-[1,3]dioxolan]-17-ol (Step B) by procedures similar to those described in Example 1, Steps B, C and D, substituting (2,3-dihydrobenzo[b][1,4]dioxin-6-yl)magnesium bromide for (3,5-difluoro-4-methylphenyl)magnesium bromide in Step C. .sup.1H NMR (400 MHz, CHLOROFORM-d) δ ppm 6.74-6.83 (1H, m),6.65-6.67 (2H, m),5.77 (1H, s), 4.32-4.39 (1H, m),4.25 (4H, s),2.72-2.83 (1H, m),2.54-2.62 (2H, m),2.30-2.51 (4H, m),2.20-2.28 (2H, m), 1.97-2.08 (2H, m), 1.90-1.97 (1H, m), 1.57-1.83 (2H, m), 1.29-1.56 (2H, m), 0.56 (3H, s). m/z (ESI, +ve ion) 448.3 (M+H).sup.+.

(24) II. Biological Evaluation

Example 5: In Vitro GR Luciferase Reporter Assay

(25) Cell Line: CHO-K1-GR-MMTV-Luc reporter cells

(26) Culture Media: DMEM (with phenol red)+10% FBS

(27) Assay Media: DMEM (without phenol red)+10% CSS

(28) Culture CHO-K1-GR-MMTV-Luc reporter cells in 15 cm plates in Culture Media at conditions less than 90% confluence.

(29) Prepare 200×DMSO 1:5 serial dilutions of control and test compounds in 96-well non-sterile V bottom plate in DMSO, 8 serial dilutions for each compound.

(30) Prepare 5× Assay Media diluted compound serial dilutions in 96-well non-sterile V bottom plate: Add 97.5 uL/well of Assay Media into 96-well then add 2.5 ul of 200× concentration of compounds and mix well.

(31) Seed cells for Antagonist Assay: 1.5×10.sup.6 CHO-K1-GR-MMTV-Luc reporter cells were seeded in a Corning 3707 flat clear bottom 384-well white TC plate in 20 ul of Assay Media containing 12.5 nM Dexamethasone (final concentration=10 nM).

(32) Add compounds: 5 ul of assay media diluted compounds were added to appropriate wells and followed a quick spin (1000 rpm, 10 sec) to bring media and cells to the bottom of plate. The plates were covered with SealMate film to avoid evaporation and placed in 37° C. incubator for approximately 18-24 hours.

(33) Read plates: Equilibrate appropriate amount of Promega OneGlo luciferase reagent to room temperature. Remove the plates from incubator and add 25 uL of OneGlo reagent/well by multiple channel pipette and read the plates with Tecan F500 luminometer within 3 minutes.

(34) The ability of the compounds disclosed herein to inhibit GR activity was quantified and the respective IC.sub.50 value was determined. Table 2 provides the cellular IC.sub.50 values of various substituted steroidal compounds disclosed herein.

(35) TABLE-US-00002 TABLE 2 GR IC.sub.50 Structure (nM) A A B B A A A A A 0 B B A A A A A A A A 00 A 01 A 02 B 03 B 04 B 05 B 06 B 07 B 08 B 09 A 0 A A A A A A A A A A 0 A A B B A A A A A A 0 A A A A A A B B B A 0 A Note: Cellular assay IC.sub.50 data are designated within the following ranges: A: ≤100 nM B: >100 nM

Example 6: In Vitro AR Agonism Assay

(36) The AR agonism assay was done in the LNAR reporter cell line which has overexpressed AR and 4XARE-Luc genes. This cell line is sensitive to even minor partial AR agonism activity in hormone-deprived media (CSS). The assays were done in RPMI (without phenol red)+10% CSS using 6000 LNAR cells/well in 384-well plates and compounds were incubated with cells in 370 C incubator for 18-24 hrs. OneGlo reagent (25 uL/well) was added and plates were read with luminometer within 3 minutes.

(37) Mifepristone shows strong partial AR agonism in concentrations as low as 10 nM, and is known to significantly promote CRPC growth both in vivo and in vitro. To gauge the AR partial agonism activity, Mifepristone is included as a standard in the assay. AR agonism assay results are quantified by determining the ratio of the maximum response of individual compounds to the maximum response of Mifepristone (arbitrarily set to 1). Values of Table 3 are reported as Relative Maximum Response to Mifepristone. This general trend of AR agonism was shown for the class of compounds described herein.

(38) TABLE-US-00003 TABLE 3 Relative Maximum Response to Structure Mifepristone A A A A A A B A A 0 A A A A A A B Note: AR agonism response assay data relative maximum response to Mifepristone is designated within the following ranges: A: ≤0.4 B: >0.4