SUBSTITUTED PYRAZOLE COMPOUNDS, COMPOSITIONS CONTAINING SAME, AND USE THEREOF

Abstract

The present invention provides substituted pyrazole compounds, compositions containing same, and use thereof. The substituted pyrazole compounds comprise a compound represented by formula (I) or a tautomer, stereoisomer, prodrug, crystalline form, pharmaceutically acceptable salt, hydrate, or solvate thereof. The compound represented by formula (I) can serve as a tissue selective androgen receptor modulator (SARM), particularly serving as a drug for treating prostate cancer and other AR-dependent conditions and diseases in which AR antagonism is desired.

Claims

1. A compound of formula (I), or a tautomer, stereoisomer, prodrug, crystal form, pharmaceutically acceptable salt, hydrate or solvate thereof: ##STR00053## wherein Y.sub.1, Y.sub.2, Y.sub.3, Y.sub.4, Y.sub.5 and Y.sub.6 are each independently selected from hydrogen, deuterium or halogen; R.sub.1, R.sub.2, R.sub.3 and R.sub.4 are each independently selected from hydrogen or deuterium; X.sub.1 and X.sub.2 are each independently selected from CH.sub.3, CD.sub.3, CHD.sub.2 or CH.sub.2D; provided that the compound comprises at least one deuterium atom.

2. The compound of claim 1, which is a compound of formula (I-1): ##STR00054## wherein Y.sub.1, Y.sub.2, Y.sub.3, Y.sub.4, Y.sub.5, Y.sub.6, R.sub.1, R.sub.2, R.sub.3, R.sub.4, X.sub.1 and X.sub.2 are as defined in claim 1; or a tautomer, stereoisomer, prodrug, crystal form, pharmaceutically acceptable salt, hydrate or solvate thereof.

3. The compound according to claim 1, which is a compound of formula (II-1): ##STR00055## wherein R.sub.1, R.sub.2, R.sub.3 and R.sub.4 are each independently selected from hydrogen or deuterium; X.sub.1 and X.sub.2 are each independently selected from CH.sub.3, CD.sub.3, CHD.sub.2 or CH.sub.2D; provided that the compound comprises at least one deuterium atom; or a tautomer, stereoisomer, prodrug, crystal form, pharmaceutically acceptable salt, hydrate or solvate thereof.

4. The compound according to claim 1, wherein X.sub.1 is CD.sub.3.

5. The compound according to claim 1, wherein X.sub.2 is CD.sub.3.

6. The compound according to claim 1, wherein R.sub.3 is deuterium.

7. The compound according to claim 1, wherein R.sub.1 and R.sub.2 are deuterium.

8. The compound according to claim 1, wherein R.sub.4 is deuterium.

9. The compound according to claim 1, which is selected from: ##STR00056## ##STR00057## ##STR00058## ##STR00059## or a tautomer, stereoisomer, prodrug, crystal form, pharmaceutically acceptable salt, hydrate or solvate thereof.

10. A pharmaceutical composition, comprising a pharmaceutically acceptable excipient and the compound according to claim 1, or a tautomer, stereoisomer, prodrug, crystal form, pharmaceutically acceptable salt, hydrate or solvate thereof.

11. (canceled)

12. A method of treating and/or preventing androgen receptor dependent diseases, comprising administering to a subject a compound of formula (I), or a tautomer, stereoisomer, prodrug, crystal form, pharmaceutically acceptable salt, hydrate or solvate thereof, or a pharmaceutical composition comprising a pharmaceutically acceptable excipient and a compound of formula (I), or a tautomer, stereoisomer, prodrug, crystal form, pharmaceutically acceptable salt, hydrate or solvate thereof, ##STR00060## wherein Y.sub.1, Y.sub.2, Y.sub.3, Y.sub.4, Y.sub.5 and Y.sub.6 are each independently selected from hydrogen, deuterium or halogen; R.sub.1, R.sub.2, R.sub.3 and R.sub.4 are each independently selected from hydrogen or deuterium; X.sub.1 and X.sub.2 are each independently selected from CH.sub.3, CD.sub.3, CHD.sub.2 or CH.sub.2D; provided that the compound comprises at least one deuterium atom.

13. The method of claim 11, wherein the androgen receptor dependent disease is prostate cancer.

Description

EXAMPLE

[0228] The present invention will be further illustrated below in conjunction with specific examples. It should be understood that these examples are only used to illustrate the present invention and not to limit the scope of the present invention. The experimental methods that do not indicate specific conditions in the following examples usually follow the conventional conditions or the conditions suggested by the manufacturer. Unless otherwise specified, parts and percentages are parts by weight and percentages by weight.

[0229] The abbreviations used herein have the following meanings:

TABLE-US-00001 DIAD diisopropyl azodicarboxylate EDCI 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride HBTU benzotriazol-N,N,N′,N′-tetramethyluronium hexafluorophosphate EDTA ethylenediamine tetraacetic acid DTT dithiothreitol Pd(OAc).sub.2 palladium acetate PPh.sub.3 triphenylphosphine Boc t-butoxycarbonyl NaH potassium hydride K.sub.2CO.sub.3 potassium carbonate NaBH.sub.4 sodium borohydride DIPEA N,N-diisopropylethylamine HCl hydrochloric acid ACN acetonitrile MeOH methanol EA ethyl acetate PE petroleum ether THF tetrahydrofuran DCM dichloromethane DMF N,N-dimethylformamide DMSO dimethyl sulfoxide Dioxane 1,4-dioxane CDCl.sub.3 deuterated chloroform

Preparation of Intermediate A1 (S)-4-O-(2-aminopropyl)-1H-pyrazol-3-yl)-2-chlorobenzonitrile

[0230] ##STR00023##

[0231] The following synthetic route was used.

##STR00024##

Step 1: Synthesis of Compound 2-chloro-4-(1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-5-yl)benzonitrile

[0232] Under nitrogen protection, Pd(OAc).sub.2 (400 mg) and PPh.sub.3 (800 mg) were added to a mixture of 4-bromo-2-chlorobenzonitrile (14.0 g, 65.1 mmol), 1-(tetrahydro-2H-pyran-2-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (21.0 g, 76.2 mmol) and potassium carbonate (18.6 g, 135 mmol) in acetonitrile (50 mL) and water (50 mL) at room temperature. The reaction was stirred at 60-70° C. for 3.5 h. The aqueous layer was separated while it was hot. The organic layer was added with 25% ammonia (2 mL), cooled to room temperature, and slowly added with water (80 mL). The reaction was stirred overnight. The solid was filtered out, washed with acetonitrile and water (20 mL, 1:1), and vacuum dried to yield 17.8 g of off-white solid. Yield: 95.6%. LC-MS (APCI): m/z=288.1 (M+1).sup.+.

Step 2: Synthesis of Compound 2-chloro-4-(1H-pyrazol-3-yl)benzonitrile

[0233] At 10±3° C., concentrated hydrochloric acid (0.54 mL) was added to a solution of 2-chloro-4-(1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-5-yl)benzonitrile (17.8 g) in methanol (70 mL), and the reaction was stirred at 10±3° C. for 2.5 h. The reaction was added with ammonia (25%, 3 mL) and then water (20 mL), stirred overnight at room temperature and then stirred at 0-5° C. for 4 h. The solid was filtered out, washed with cold water and methanol (3:1, 40 mL), and vacuum dried to obtain 9.8 g of off-white solid. Yield: 78%. LC-MS (APCI): m/z=204.1 (M+1).sup.+.

Step 3: Synthesis of Intermediate A1

[0234] At 10±5° C., DIAD (2.97 mL, 14.8 mmol) was slowly added dropwise to a solution of 2-chloro-4-(1H-pyrazol-3-yl)benzonitrile (1.5 g, 7.4 mmol), N-Boc-L-alaninol (2.65 g, 14.8 mmol) and PPh.sub.3 (3.96 g, 14.8 mmol) in anhydrous ethyl acetate (20 mL) under nitrogen protection. After the addition, the reaction was stirred overnight at room temperature. Concentrated hydrochloric acid (3.1 mL) was slowly added, with the temperature of the reaction system not exceeding 35° C. during the addition process. After the addition was complete, the reaction was carried out at 45±5° C. until the reaction was complete. The reaction was adjusted to alkaline pH with concentrated ammonia, extracted with DCM, and dried with anhydrous sodium sulfate. 1.5 g of white solid was obtained after filtration, concentration under reduced pressure, and purification of the concentrated liquid by column chromatography (DCM/MeOH=8%). Yield: 78%. LC-MS (APCI): m/z=261.2 (M+1).sup.+.

Preparation of Intermediate A2 (S)-4-(1-(2-aminopropyl-1,1-d.SUB.2.)-1H-pyrazol-3-yl)-2-chlorobenzonitrile

[0235] ##STR00025##

[0236] The following synthetic route was used.

##STR00026##

Step 1: Synthesis of Compound tert-butyl (S)-(1-hydroxyprop-2-yl-1,1-d.SUB.2.)carbamate

[0237] Under ice bath, LiAlD.sub.4 (1.0 g, 23.8 mmol) was added in portions to Boc-L-alanine methyl ester (4.03 g, 19.8 mmol) in anhydrous THF (40 ml), and reacted at room temperature for 1 hr. It was quenched with NaSO.sub.4.10H.sub.2O, filtered, and washed twice with THF (30 mL×2), and the filtrate was rotation-dried to obtain 3.2 g of off-white solid. Yield: 91%. LC-MS (APCI): m/z=178.2 (M+1).sup.+, .sup.1H NMR (500 MHz, CDCl.sub.3) δ 4.66 (s, 1H), 3.77 (s, 1H), 2.61 (s, 1H), 1.45 (s, 9H), 1.15 (d, J=6.8 Hz, 3H).

Step 2: Synthesis of Intermediate A2

[0238] At 10±5° C., DIAD (2.97 mL, 14.8 mmol) was slowly added dropwise to a solution of 2-chloro-4-(1H-pyrazol-3-yl)benzonitrile (1.5 g, 7.4 mmol), tert-butyl (S)-(1-hydroxyprop-2-yl-1,1-d.sub.2)carbamate (2.68 g, 14.8 mmol) and PPh.sub.3 (3.96 g, 14.8 mmol) in anhydrous ethyl acetate (20 mL) under nitrogen protection. After the addition, the reaction was stirred overnight at room temperature. Concentrated hydrochloric acid (3.1 mL) was slowly added, with the temperature of the reaction system not exceeding 35° C. during the addition process. After the addition was complete, the reaction was carried out at 45±5° C. until the reaction was complete. The reaction was adjusted to alkaline pH with concentrated ammonia, extracted with DCM, and dried with anhydrous sodium sulfate. 1.52 g of white solid was obtained after filtration, concentration under reduced pressure, and purification of the concentrated liquid by column chromatography (DCM/MeOH=8%). Yield: 79%. LC-MS (APCI): m/z=263.2 (M+1).sup.+.

Preparation of Intermediate A3 (S)-4-(1-(2-aminopropyl-2-d)-1H-pyrazol-3-yl)-2-chlorobenzonitrile

[0239] ##STR00027##

[0240] The following synthetic route was used.

##STR00028##

Step 1: Synthesis of Compound 2-chloro-4-(1-(2-oxopropyl)-1H-pyrazol-3-yl)benzonitrile

[0241] Under ice bath, NaH (600 mg, 15 mmol) was added in portions to a solution of 2-chloro-4-(1H-pyrazol-3-yl)benzonitrile (2.0 g, 9.85 mmol) in DMF (20 ml), and reacted at room temperature for 0.5 h. The above system was added with bromoacetone (2.56 g, 18.8 mmol), and reacted at room temperature for 2 h. The system was quenched with water, extracted with EA (100 mL×3), and dried with anhydrous sodium sulfate. Filtration, concentration under reduced pressure, and purification of the concentrated liquid by column chromatography (PE/EA=22%) provided 1.1 g of white solid. Yield: 43%. LC-MS (APCI): m/z=260.1 (M+1).sup.+.

Step 2: Synthesis of Compound N—((S)-1-(3-(3-chloro-4-cyanophenyl)-1H-pyrazol-1-yl)prop-2-yl-2-d)-2-methylpropan-2-sulfinamide

[0242] At room temperature, (S)-tert-butylsulfinamide (185 mg, 1.53 mmol) and tetraisopropyl titanate (789 mg, 2.78 mmol) were added to a solution of 2-chloro-4-(1-(2-oxopropyl)-1H-pyrazol-3-yl)benzonitrile (360 mg, 1.39 mmol) in THF (10 mL), and reacted at 72° C. overnight. At −20° C., NaBD.sub.4 (168 mg, 5.56 mmol) was added in portions, and reacted for 2 h at room temperature. It was quenched with water, extracted with EA (100 mL×3), and dried with anhydrous sodium sulfate. Filtration, concentration under reduced pressure, and purification of the concentrated liquid by column chromatography (PE/EtOAc=64%) provided 360 mg of white solid. Yield: 65%. LC-MS (APCI): m/z=364.1 (M+1).sup.+.

Step 3: Synthesis of Intermediate A3

[0243] At room temperature, HCl (4M, MeOH solution) (3 ml) was added to a solution of N—((S)-1-(3-(3-chloro-4-cyanophenyl)-1H-pyrazol-1-yl)prop-2-yl)-2-methylpropan-2-sulfinamide (360 mg, 0.99 mmol) in methanol (5 ml), and reacted at room temperature for 1 h. Rotation-drying and purification by column chromatography (DCM/MeOH=8%) provided 116 mg of white solid. Yield: 45%. LC-MS (APCI): m/z=262.1 (M+1).sup.+.

Preparation of Intermediate A4 (S)-4-(1-(2-aminopropyl-1,1,3,3,3-d.SUB.5.)-1H-pyrazol-3-yl)-2-chlorobenzonitrile

[0244] ##STR00029##

[0245] The following synthetic route was used.

##STR00030##

Step 1: Synthesis of Compound 2-chloro-4-(1-(2-oxopropyl-1,1,3,3,3-d.SUB.5.)-1H-pyrazol-3-yl)benzonitrile

[0246] At room temperature, tetrahydropyrrole (5 mg) was added to a solution of 2-chloro-4-(1-(2-oxopropyl)-1H-pyrazol-3-yl)benzonitrile (1 g, 3.85 mmol) in a mixture of 1,4-dioxane (15 mL) and D.sub.2O (15 mL), and reacted for 72 h at room temperature. Rotation drying, concentration under reduced pressure, and purification of the concentrated liquid by column chromatography (PE/EA=23%) provided 850 mg of white solid. Yield: 86%. LC-MS (APCI): m/z=265.1 (M+1)+, .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ 8.08 (d, J=1.2 Hz, 1H), 8.00-7.90 (m, 2H), 7.77 (d, J=2.4 Hz, 1H), 7.01 (d, J=2.4 Hz, 1H).

Step 2: Synthesis of Intermediate A4

[0247] At room temperature, (S)-tert-butylsulfinamide (358 mg, 2.96 mmol) and tetraisopropyl titanate (1.53 g, 5.38 mmol) were added to a solution of 2-chloro-4-(1-(2-oxopropyl-1,1,3,3,3-d.sub.5)-1H-pyrazol-3-yl)benzonitrile (710 mg, 2.69 mmol) in THF (10 mL), and reacted overnight at 72° C. At −20° C., NaBD.sub.4 (500 mg, 13.45 mmol) was added in portions, and reacted for 2 h at room temperature. It was quenched with water, extracted with EA (100 mL×3), and dried with anhydrous sodium sulfate. Filtration, concentration under reduced pressure, and purification of the concentrated liquid by column chromatography (PE/EA=64%) provided 750 mg of white solid.

[0248] At room temperature, HCl (4M, MeOH solution) (3 mL) was added to a solution of the above product in methanol (5 mL), and reacted for 1 h at room temperature. Rotation drying and purification by column chromatography (DCM/MeOH=8%) provided 420 mg of white solid. Yield: 45%. LC-MS (APCI): m/z=266.1 (M+1).sup.+.

Preparation of Intermediate B 5-acetyl-1H-pyrazole-3-carboxylic Acid

[0249] ##STR00031##

[0250] The following synthetic route was used.

##STR00032##

Step 1: Synthesis of Compound 5-acetyl-1H-pyrazole-3-carboxylic acid ethyl ester

[0251] At room temperature, ethyl azidoacetate (12.5 g, 110 mmol) was slowly added dropwise to a solution of 3-ynyl-2-butanone (5 g, 73 mmol) in water (80 mL). The reaction was stirred at room temperature for 4 hours, and the solid was filtered out to obtain 11 g of light yellow solid. Yield: 81%. LC-MS (APCI): m/z=183.2, .sup.1H NMR (300 MHz, CDCl.sub.3) δ 11.69 (s, 1H), 7.33 (s, 1H), 4.43 (q, J=7.1 Hz, 2H), 2.61 (s, 3H), 1.41 (t, J=7.1 Hz, 3H).

Step 2: Synthesis of Intermediate B

[0252] At room temperature, NaOH (4.4 g, 108 mmol) was added to a solution of 5-acetyl-1H-pyrazole-3-carboxylic acid ethyl ester (4.0 g, 22 mmol) in a mixture of methanol (20 mL) and water (20 mL). The reaction was stirred at room temperature for 2 hours, and adjusted to pH 2 with concentrated hydrochloric acid. The solid was filtered out to obtain 3.2 g of off-white solid. Yield: 94%. LC-MS (APCI): m/z=155.1.

Example 1

Preparation of N—((S)-1-(3-(3-chloro-4-cyanophenyl)-1H-pyrazol-1-yl)prop-2-yl)-5-(1-hydroxylethyl-1-d)-1H-pyrazole-3-carboxamide

[0253] ##STR00033##

[0254] The following synthetic route was used.

##STR00034##

Step 1: Synthesis of Compound (S)-5-acetyl-N-(1-(3-(3-chloro-4-cyanophenyl)-1H-pyrazol-1-yl)prop-2-yl)-1H-pyrazole-3-carboxamide

[0255] At room temperature, HBTU (1.53 g, 4.04 mmol), EDCI (775 mg, 4.04 mmol) and Intermediate A1 (700 mg, 2.69 mmol) were added to a solution of Intermediate B (500 mg, 3.23 mmol) and DIPEA (748 mg, 5.38 mmol) in anhydrous DCM (10 mL), and the reaction was stirred at room temperature overnight. The reaction was quenched with water, extracted with EA (100×3), and dried with anhydrous sodium sulfate. Filtration, concentration under reduced pressure, and purification of the concentrated liquid by column chromatography (PE/EA=80%) provided 534 mg of white solid. Yield: 50%. LC-MS (APCI): m/z=397.1 (M+1).sup.+.

Step 2: Synthesis of Compound N—((S)-1-(3-(3-chloro-4-cyanophenyl)-1H-pyrazol-1-yl)prop-2-yl)-5-(1-hydroxylethyl-1-d)-1H-pyrazole-3-carboxamide

[0256] At room temperature, NaBD.sub.4 (32 mg, 0.76 mmol) was added to a solution of (S)-5-acetyl-N-(1-(3-(3-chloro-4-cyanophenyl)-1H-pyrazol-1-yl)prop-2-yl)-1H-pyrazole-3-carboxamide (150 mg, 0.38 mmol) in anhydrous CD.sub.3CD.sub.2OD (5 mL), and the reaction was stirred at room temperature for 2.5 h. The reaction was quenched by H.sub.2O (20 mL) and HCl (0.5M, 0.3 mL). CD.sub.3CD.sub.2OD was removed under reduced pressure, and the residue was dissolved in DCM, washed with 1M sodium bicarbonate solution and water, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Purification of the concentrated liquid by column chromatography (DCM/MeOH=6%) provided 56 mg of white solid. Yield: 51.85%, purity: 99.46%. LC-MS (APCI): m/z=400.1 (M+1).sup.+, .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ 13.04 (s, 1H), 8.21 (d, J=7.7 Hz, 1H), 8.07 (s, 1H), 7.98 (s, 2H), 7.80 (d, J=2.2 Hz, 1H), 6.93 (d, J=2.2 Hz, 1H), 6.38 (s, 1H), 5.41 (s, 1H), 4.48-4.28 (m, 3H), 1.36 (d, J=6.3 Hz, 3H), 1.09 (d, J=5.7 Hz, 3H).

Example 2

Preparation of N—((S)-1-(3-(3-chloro-4-cyanophenyl)-1H-pyrazol-1-yl)prop-2-yl)-5-(1-hydroxylethyl-2,2,2-d.SUB.3.)-1H-pyrazole-3-carboxamide

[0257] ##STR00035##

[0258] The following synthetic route was used.

##STR00036##

Step 1: Synthesis of Compound (S)-5-(acetyl-d.SUB.3.)-N-(1-(3-(3-chloro-4-cyanophenyl)-1H-pyrazol-1-yl)prop-2-yl)-1H-pyrazole-3-carboxamide

[0259] At room temperature, tetrahydropyrrole (5 mg) was added to a solution of (S)-5-acetyl-N-(1-(3-(3-chloro-4-cyanophenyl)-1H-pyrazol-1-yl)prop-2-yl)-1H-pyrazole-3-carboxamide (400 mg, 1.0 mmol) in a mixture of 1,4-dioxane (10 mL) and D.sub.2O (10 mL), and reacted for 72 h at room temperature. Rotation drying, concentration under reduced pressure, and purification of the concentrated liquid by column chromatography (PE/EA=81%) provided 380 mg of white solid. Yield: 95%. LC-MS (APCI): m/z=400.1 (M+1).sup.+, .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ 14.18 (d, J=39.3 Hz, 1H), 8.51 (s, 1H), 8.00 (d, J=28.9 Hz, 2H), 7.82 (d, J=2.3 Hz, 1H), 7.62 (s, 1H) 7.32 (s, 1H), 6.94 (d, J=2.1 Hz, 1H), 4.44-4.29 (m, 3H), 1.19-1.13 (m, 3H).

Step 2: Synthesis of Compound N—((S)-1-(3-(3-chloro-4-cyanophenyl)-1H-pyrazol-1-yl)prop-2-yl)-5-(1-hydroxylethyl-2,2,2-d.SUB.3.)-1H-pyrazole-3-carboxamide

[0260] At room temperature, NaBH.sub.4 (28 mg, 74 mmol) was added to (S)-5-(acetyl-d.sub.3)-N-(1-(3-(3-chloro-4-cyanophenyl)-1H-pyrazol-1-yl)prop-2-yl)-1H-pyrazole-3-carboxamide (150 mg, 0.37 mmol) in absolute ethanol (5 mL), and the reaction was stirred at room temperature for 2.5 h. The reaction was quenched by H.sub.2O (10 mL) and HCl (0.5M, 0.3 mL). Ethanol was removed under reduced pressure, and the residue was dissolved in DCM, washed with 1M sodium bicarbonate solution and water, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Purification of the concentrated liquid by column chromatography (DCM/MeOH=6%) provided 78 mg of white solid. Yield: 52%, purity: 99.44%. m/z=402.1 (M+1)+, .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ 13.04 (s, 1H), 8.21 (d, J=7.7 Hz, 1H), 8.07 (s, 1H), 7.98 (s, 2H), 7.80 (d, J=2.2 Hz, 1H), 6.93 (d, J=2.2 Hz, 1H), 6.38 (s, 1H), 5.41 (s, 1H), 4.77 (s, 1H), 4.48-4.40 (s, 3H), 1.09 (d, J=5.7 Hz, 3H).

Example 3

Preparation of N—((S)-1-(3-(3-chloro-4-cyanophenyl)-1H-pyrazol-1-yl)prop-2-yl)-5-(1-hydroxylethyl-1,2,2,2-d.SUB.4.)-1H-pyrazole-3-carboxamide

[0261] ##STR00037##

[0262] The following synthetic route was used.

##STR00038##

[0263] At room temperature, NaBD.sub.4 (31 mg, 74 mmol) was added to (S)-5-(acetyl-d.sub.3)-N-(1-(3-(3-chloro-4-cyanophenyl)-1H-pyrazol-1-yl)prop-2-yl)-1H-pyrazole-3-carboxamide (150 mg, 0.37 mmol) in anhydrous CD.sub.3CD.sub.2OD (5 mL), and the reaction was stirred at room temperature for 2.5 h. The reaction was quenched by H.sub.2O (10 mL) and HCl (0.5M, 0.3 mL). CD.sub.3CD.sub.2OD was removed under reduced pressure, and the residue was dissolved in DCM, washed with 1M sodium bicarbonate solution and water, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Purification of the concentrated liquid by column chromatography (DCM/MeOH=6%) provided 69 mg of white solid. Yield: 46%, purity: 99.37%. LC-MS (APCI): m/z=403.1 (M+1).sup.+, .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ 13.04 (s, 1H), 8.21 (d, J=7.7 Hz, 1H), 8.07 (s, 1H), 7.98 (s, 2H), 7.80 (d, J=2.2 Hz, 1H), 6.93 (d, J=2.2 Hz, 1H), 6.38 (s, 1H), 5.41 (s, 1H), 4.48-4.40 (s, 3H), 1.09 (d, J=5.7 Hz, 3H).

Example 4

Preparation of N—((S)-1-(3-(3-chloro-4-cyanophenyl)-1H-pyrazol-1-yl)prop-2-yl-1,1-d.SUB.2.)-5-(1-hydroxyethyl)-1H-pyrazole-3-carboxamide

[0264] ##STR00039##

[0265] The following synthetic route was used.

##STR00040##

Step 1: Synthesis of Compound (S)-5-acetyl-N-(1-(3-(3-chloro-4-cyanophenyl)-1H-pyrazol-1-yl)prop-2-yl-1,1-d.SUB.2.)-1H-pyrazole-3-carboxamide

[0266] At room temperature, HBTU (1215 mg, 3.21 mmol), EDCI (616 mg, 3.21 mmol) and Intermediate A2 (560 mg, 2.14 mmol) were added to a solution of Intermediate B (658 mg, 4.28 mmol) and DIPEA (594 mg, 4.28 mmol) in anhydrous DCM (10 mL), and the reaction was stirred at room temperature overnight. The reaction was quenched with water, extracted with EA (50×3), and dried with anhydrous sodium sulfate. Filtration, concentration under reduced pressure, and purification of the concentrated liquid by column chromatography (PE/EtOAc=81%) provided 500 mg of white solid. Yield: 58.2%. LC-MS (APCI): m/z=399.1 (M+1).sup.+.

Step 2: Synthesis of Compound N—((S)-1-(3-(3-chloro-4-cyanophenyl)-1H-pyrazol-1-yl)prop-2-yl-1,1-d.SUB.2.)-5-(1-hydroxyethyl)-1H-pyrazole-3-carboxamide

[0267] At room temperature, NaBH.sub.4 (48 mg, 1.25 mmol) was added to (S)-5-acetyl-N-(1-(3-(3-chloro-4-cyanophenyl)-1H-pyrazol-1-yl)prop-2-yl-1,1-d.sub.2)-1H-pyrazole-3-carboxamide (250 mg, 0.628 mmol) in anhydrous ethanol (5 mL), and the reaction was stirred at room temperature for 2.5 h. The reaction was quenched by H.sub.2O (10 mL) and HCl (0.5M, 0.3 mL). Ethanol was removed under reduced pressure, and the residue was dissolved in DCM, washed with 1M sodium bicarbonate solution and water, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Purification of the concentrated liquid by column chromatography (DCM/MeOH=6%) provided 90 mg of white solid. Yield: 36%, purity: 92.67%. LC-MS (APCI): m/z=401.1 (M+1).sup.+, .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ 13.04 (s, 1H), 8.21 (d, J=7.7 Hz, 1H), 8.07 (s, 1H), 7.98 (s, 2H), 7.80 (d, J=2.2 Hz, 1H), 6.93 (d, J=2.2 Hz, 1H), 6.38 (s, 1H), 5.41 (s, 1H), 5.09 (s, OH), 4.77 (s, 1H), 4.48-4.40 (s, 1H), 1.36 (d, J=6.3 Hz, 3H), 1.09 (d, J=5.7 Hz, 3H).

Example 5

Preparation of N—((S)-1-(3-(3-chloro-4-cyanophenyl)-1H-pyrazol-1-yl)prop-2-yl-1,1-d.SUB.2.)-5-(1-hydroxyethyl-1-d)-1H-pyrazole-3-carboxamide

[0268] ##STR00041##

[0269] The following synthetic route was used.

##STR00042##

[0270] At room temperature, NaBD.sub.4 (53 mg, 1.25 mmol) was added to (S)-5-acetyl-N-(1-(3-(3-chloro-4-cyanophenyl)-1H-pyrazol-1-yl)prop-2-yl-1,1-d.sub.2)-1H-pyrazole-3-carboxamide (250 mg, 0.628 mmol) in anhydrous CD.sub.3CD.sub.2OD (5 mL), and the reaction was stirred at room temperature for 2.5 h. The reaction was quenched by H.sub.2O (10 mL) and HCl (0.5M, 0.3 mL). Ethanol was removed under reduced pressure, and the residue was dissolved in DCM, washed with 1M sodium bicarbonate solution and water, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Purification of the concentrated liquid by column chromatography (DCM/MeOH=6%) provided 135 mg of white solid. Yield: 54%, purity: 97.78%. LC-MS (APCI): m/z=402.1 (M+1).sup.+, .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ 13.04 (s, 1H), 8.21 (d, J=7.7 Hz, 1H), 8.07 (s, 1H), 7.98 (s, 2H), 7.80 (d, J=2.2 Hz, 1H), 6.93 (d, J=2.2 Hz, 1H), 6.38 (s, 1H), 5.41 (s, 1H), 5.09 (s, OH), 4.48-4.40 (s, 1H), 1.36 (d, J=6.3 Hz, 3H), 1.09 (d, J=5.7 Hz, 3H).

Example 6

Preparation of N—((S)-1-(3-(3-chloro-4-cyanophenyl)-1H-pyrazol-1-yl)prop-2-yl-1,1-d.SUB.2.)-5-(1-hydroxyethyl-2,2,2-d.SUB.3.)-1H-pyrazole-3-carboxamide

[0271] ##STR00043##

[0272] The following synthetic route was used.

##STR00044##

Step 1: Synthesis of Compound (S)-5-(acetyl-d.SUB.3.)-N-(1-(3-(3-chloro-4-cyanophenyl)-1H-pyrazol-1-yl)prop-2-yl-1,1-d.SUB.2.)-1H-pyrazole-3-carboxamide

[0273] At room temperature, tetrahydropyrrole (5 mg) was added to a solution of (S)-5-(acetyl)-N-(1-(3-(3-chloro-4-cyanophenyl)-1H-pyrazol-1-yl)prop-2-yl-1,1-d.sub.2)-1 H-pyrazole-3-carboxamide (400 mg, 1.0 mmol) in a mixture of 1,4-dioxane (10 mL) and D.sub.2O (10 mL), and reacted for 72 h at room temperature. Rotation drying, concentration under reduced pressure, and purification of the concentrated liquid by column chromatography (PE/EA=81%) provided 380 mg of white solid. Yield: 95%. LC-MS (APCI): m/z=402.1 (M+1).sup.+, .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ 14.18 (d, J=39.3 Hz, 1H), 8.51 (s, 1H), 8.00 (d, J=28.9 Hz, 2H), 7.82 (d, J=2.3 Hz, 1H), 7.62 (s, 1H), 7.32 (s, 1H), 6.94 (d, J=2.1 Hz, 1H), 4.44 (s, 1H), 1.19-1.13 (m, 3H).

Step 2: Synthesis of Compound N—((S)-1-(3-(3-chloro-4-cyanophenyl)-1H-pyrazol-1-yl)prop-2-yl-1,1-d.SUB.2.)-5-(1-hydroxyethyl-2,2,2-d.SUB.3.)-1H-pyrazole-3-carboxamide

[0274] At room temperature, NaBH.sub.4 (28 mg, 74 mmol) was added to (S)-5-(acetyl-d.sub.3)-N-(1-(3-(3-chloro-4-cyanophenyl)-1H-pyrazol-1-yl)prop-2-yl-1,1-d.sub.2)-1H-pyrazole-3-carboxamide (150 mg, 0.37 mmol) in absolute ethanol (5 mL), and the reaction was stirred at room temperature for 2.5 h. The reaction was quenched by H.sub.2O (10 mL) and HCl (0.5M, 0.3 mL). Ethanol was removed under reduced pressure, and the residue was dissolved in DCM, washed with 1M sodium bicarbonate solution and water, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Purification of the concentrated liquid by column chromatography (DCM/MeOH=6%) provided 100 mg of white solid. Yield: 67%, purity: 99.6%. LC-MS (APCI): m/z=402.1 (M+1).sup.+, .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ 13.04 (s, 1H), 8.21 (d, J=7.7 Hz, 1H), 8.07 (s, 1H), 7.98 (s, 2H), 7.80 (d, J=2.2 Hz, 1H), 6.93 (d, J=2.2 Hz, 1H), 6.38 (s, 1H), 5.41 (s, 1H), 4.77 (s, 1H), 4.48-4.40 (s, 1H), 1.09 (d, J=5.7 Hz, 3H).

Example 7

Preparation of N—((S)-1-(3-(3-chloro-4-cyanophenyl)-1H-pyrazol-1-yl)prop-2-yl-1,1-d.SUB.2.)-5-(1-hydroxyethyl-1,2,2,2-d.SUB.4.)-1H-pyrazole-3-carboxamide

[0275] ##STR00045##

[0276] The following synthetic route was used.

##STR00046##

[0277] At room temperature, NaBD.sub.4 (31 mg, 74 mmol) was added to (S)-5-(acetyl-d.sub.3)-N-(1-(3-(3-chloro-4-cyanophenyl)-1H-pyrazol-1-yl)prop-2-yl-1,1-d.sub.2)-1H-pyrazole-3-carboxamide (150 mg, 0.37 mmol) in anhydrous CD.sub.3CD.sub.2OD (5 mL), and the reaction was stirred at room temperature for 2.5 h. The reaction was quenched by H.sub.2O (10 mL) and HCl (0.5M, 0.3 mL). Ethanol was removed under reduced pressure, and the residue was dissolved in DCM, washed with 1M sodium bicarbonate solution and water, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Purification of the concentrated liquid by column chromatography (DCM/MeOH=6%) provided 104 mg of white solid. Yield: 69%, purity: 97.89%. LC-MS (APCI): m/z=402.1 (M+1).sup.+, .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ 13.04 (s, 1H), 8.21 (d, J=7.7 Hz, 1H), 8.07 (s, 1H), 7.98 (s, 2H), 7.80 (d, J=2.2 Hz, 1H), 6.93 (d, J=2.2 Hz, 1H), 6.38 (s, 1H), 5.41 (s, 1H), 4.48-4.40 (s, 1H), 1.09 (d, J=5.7 Hz, 3H).

Example 8

Preparation of N—((S)-1-(3-(3-chloro-4-cyanophenyl)-1H-pyrazol-1-yl)prop-2-yl-2-d)-5-(1-hydroxyethyl)-1H-pyrazole-3-carboxamide

[0278] ##STR00047##

[0279] The following synthetic route was used.

##STR00048##

Step 1: Synthesis of Compound (S)-5-(acetyl)-N-(1-(3-(3-chloro-4-cyanophenyl)-1H-pyrazol-1-yl)prop-2-yl-2-d)-1H-pyrazole-3-carboxamide

[0280] At room temperature, HBTU (253 mg, 0.66 mmol), EDCI (128 mg, 0.66 mmol) and Intermediate A3 (116 mg, 0.44 mmol) were added to a solution of Intermediate B (136 mg, 0.88 mmol) and DIPEA (124 mg, 0.88 mmol) in anhydrous DCM (10 mL), and the reaction was stirred at room temperature overnight. The reaction was quenched with water, extracted with EA (30×3), and dried with anhydrous sodium sulfate. Filtration, concentration under reduced pressure, and purification of the concentrated liquid by column chromatography (PE/EA=83%) provided 150 mg of white solid. Yield: 85%. LC-MS (APCI): m/z=398.1 (M+1).sup.+.

Step 2: Synthesis of Compound N—((S)-1-(3-(3-chloro-4-cyanophenyl)-1H-pyrazol-1-yl)prop-2-yl-2-d)-5-(1-hydroxyethyl)-1H-pyrazole-3-carboxamide

[0281] At room temperature, NaBH.sub.4 (29 mg, 7.2 mmol) was added to (S)-5-(acetyl)-N-(1-(3-(3-chloro-4-cyanophenyl)-1H-pyrazol-1-yl)prop-2-yl-2-d)-1H-pyrazole-3-carboxamide (150 mg, 0.38 mmol) in absolute ethanol (5 mL), and the reaction was stirred at room temperature for 2.5 h. The reaction was quenched by H.sub.2O (10 mL) and HCl (0.5M, 0.3 mL). Ethanol was removed under reduced pressure, and the residue was dissolved in DCM, washed with 1M sodium bicarbonate solution and water, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Purification of the concentrated liquid by column chromatography (DCM/MeOH=6%) provided 46 mg of white solid. Yield: 31%, purity: 97.81%. LC-MS (APCI): m/z=400.1 (M+1).sup.+, .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ 13.04 (s, 1H), 8.21 (d, J=7.7 Hz, 1H), 8.07 (s, 1H), 7.98 (s, 2H), 7.80 (d, J=2.2 Hz, 1H), 6.93 (d, J=2.2 Hz, 1H), 6.38 (s, 1H), 5.41 (s, 1H), 4.77 (s, 1H), 4.45-4.40 (s, 2H), 1.36 (d, J=6.3 Hz, 3H), 1.09 (d, J=5.7 Hz, 3H).

Example 9

Preparation of N—((S)-1-(3-(3-chloro-4-cyanophenyl)-1H-pyrazol-1-yl)prop-2-yl-1,1,3,3,3-d.SUB.5.)-5-(1-hydroxyethyl)-1H-pyrazole-3-carboxamide

[0282] ##STR00049##

[0283] The following synthetic route was used.

##STR00050##

Step 1: Synthesis of Compound (S)-5-(acetyl)-N-(1-(3-(3-chloro-4-cyanophenyl)-1H-pyrazol-1-yl)prop-2-yl-1,1,3,3,3-d.SUB.5.)-1H-pyrazole-3-carboxamide

[0284] At room temperature, HBTU (921 mg, 2.33 mmol), EDCI (446 mg, 2.33 mmol) and Intermediate A4 (410 mg, 1.55 mmol) were added to a solution of Intermediate B (284 mg, 1.86 mmol) and DIPEA (430 mg, 2.33 mmol) in anhydrous DCM (10 mL), and the reaction was stirred at room temperature overnight. The reaction was quenched with water, extracted with EA (30×3), and dried with anhydrous sodium sulfate. Filtration, concentration under reduced pressure, and purification of the concentrated liquid by column chromatography (PE/EtOAc=83%) provided 300 mg of white solid. Yield: 85%. LC-MS (APCI): m/z=402.1 (M+1).sup.+.

Step 2: Synthesis of Compound N—((S)-1-(3-(3-chloro-4-cyanophenyl)-1H-pyrazol-1-yl)prop-2-yl-1,1,3,3,3-d.SUB.5.)-5-(1-hydroxyethyl)-1H-pyrazole-3-carboxamide

[0285] At room temperature, NaBH.sub.4 (29 mg, 7.2 mmol) was added to (S)-5-(acetyl)-N-(1-(3-(3-chloro-4-cyanophenyl)-1H-pyrazol-1-yl)prop-2-yl-1,1,3,3,3-d.sub.5)-1H-pyrazole-3-carboxamide (150 mg, 0.38 mmol) in absolute ethanol (5 mL), and the reaction was stirred at room temperature for 2.5 h. The reaction was quenched by H.sub.2O (10 mL) and HCl (0.5M, 0.3 mL). Ethanol was removed under reduced pressure, and the residue was dissolved in DCM, washed with 1M sodium bicarbonate solution and water, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Purification of the concentrated liquid by column chromatography (DCM/MeOH=6%) provided 56 mg of white solid. Yield: 37%, purity: 97.81%. LC-MS (APCI): m/z=404.1 (M+1).sup.+, .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ 13.04 (s, 1H), 8.21 (d, J=7.7 Hz, 1H), 8.07 (s, 1H), 7.98 (s, 2H), 7.80 (d, J=2.2 Hz, 1H), 6.93 (d, J=2.2 Hz, 1H), 6.38 (s, 1H), 5.41 (s, 1H), 4.77 (s, 1H), 4.45-4.40 (s, 1H), 1.36 (d, J=6.3 Hz, 3H).

Example 10

Preparation of N—((S)-1-(3-(3-chloro-4-cyanophenyl)-1H-pyrazol-1-yl)prop-2-yl-1,1,3,3,3-d.SUB.5.)-5-(1-hydroxyethyl-1-d)-1H-pyrazole-3-carboxamide

[0286] ##STR00051##

[0287] The following synthetic route was used.

##STR00052##

[0288] At room temperature, NaBD.sub.4 (31 mg, 7.2 mmol) was added to (S)-5-(acetyl)-N-(1-(3-(3-chloro-4-cyanophenyl)-1H-pyrazol-1-yl)prop-2-yl-1,1,3,3,3-d.sub.5)-1H-pyrazole-3-carboxamide (150 mg, 0.38 mmol) in anhydrous CD.sub.3CD.sub.2OD (5 mL), and the reaction was stirred at room temperature for 2.5 h. The reaction was quenched by H.sub.2O (10 mL) and HCl (0.5M, 0.3 mL). Ethanol was removed under reduced pressure, and the residue was dissolved in DCM, washed with 1M sodium bicarbonate solution and water, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Purification of the concentrated liquid by column chromatography (DCM/MeOH=6%) provided 48 mg of white solid. Yield: 32%. LC-MS (APCI): m/z=405.1 (M+1).sup.+, .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ 13.04 (s, 1H), 8.21 (d, J=7.7 Hz, 1H), 8.07 (s, 1H), 7.98 (s, 2H), 7.80 (d, J=2.2 Hz, 1H), 6.93 (d, J=2.2 Hz, 1H), 6.38 (s, 1H), 5.41 (s, 1H), 4.45-4.40 (s, 1H), 1.36 (d, J=6.3 Hz, 3H).

[0289] Biological Activity Test

[0290] (1) Inhibition of Cellular PSA Protein Secretion

[0291] Experimental Steps:

1. Replacing the original medium with a medium containing 10% Charcoal Stripped FBS, and starving the cells in the culture flask for 24 hours;
2. Digesting the cells, counting them, seeding LNcaP cells in a 96-well plate at 10,000/well and incubating them overnight;
3. Adding DHT and the compound to the existing medium according to the set concentration, the final concentration of DHT being 1 nM and the initial concentration of compound being 50000 nM, 5-fold dilution, 8 concentration gradients, and incubating them for 48 hours; and 4. Collecting the cell culture supernatant, and detecting the PSA protein level according to the ELISA kit instructions.

[0292] According to the inhibition rate at each concentration, the IC50 was calculated with GraphPad Prism, and the results were shown in the table below.

[0293] It can be seen from the above table that the compounds of the present invention have a high inhibitory activity on cellular PSA protein, and thus can be used as medicines for treating prostate cancer.

[0294] (2) Androgen Receptor AR Affinity Test

[0295] The cytoplasm extracted from LNCap cells and commercial radioisotopes were used for AR affinity experiments.

[0296] The highest concentration of the drug to be tested was 1 μM, with 4-fold dilution and 8 concentrations. 1 μL of drug solution was added to each well of a 96-well plate (Agilent, 5042-1385) in duplicate. Each well was added with 100 μL of cytoplasm extracted from LNCap cells (600 ug/well), and 100 μL of radioisotope-labeled 3H-methyltrienone (final concentration 1.0 nM, PerkinElmer, Cat: NET590250UC, Lot: 2133648). The plate was sealed and shaken at 300 rpm at 4° C. for 24 h. 100 μL of radioligand adsorption buffer (Tris-HCl (10 mM), pH 7.4; EDTA (1.5 mM); DTT (1 mM); 0.25% activated carbon; 0.0025% dextran) was added, followed by shaking for 15 min at 4° C., and then centrifugation at 4° C., 3000 rpm for 30 min. 150 μL of the supernatant was transferred to a 6 ml Scint-tube (PerkinElmer, 6000192), and 2 mL of Ultima Gold cocktail (PerkinElmer, Cat: 6013329, Lot: 77-16371) was added. Tri-Carb 2910 TR (PerkinElmer) was used for isotope labeling reading, and the inhibition rate was calculated. Data was analyzed with GraphPad Prism 5.0 software, and fitted using nonlinear curve regression to produce the dose-effect curve and thereby calculate the IC.sub.50 value.

[0297] The compounds of the present invention were tested in the above AR affinity experiment. Compared with the non-deuterated compound Darolutamide, the compounds of the present invention have a comparable affinity and have an antagonistic effect on AR.

[0298] (3) DU145 Cell Proliferation Experiment (Negative Control)

[0299] The cell concentration was adjusted. 50 μL cell suspension was added to a 384-well plate, and incubated overnight at 37° C. and 5% CO.sub.2. Tecan D300E program was set up. The drug was added using a Tecan D300E instrument, the highest concentration of the drug to be tested being 10 μM, with a 3-fold gradient dilution, 9 concentrations, in duplicate, and the incubation was continued for 72 hours. The 384-well plate was taken out and equilibrated at room temperature for 30 minutes. Each well was added with 30 μL of CTG (Promega, G7573) reagent, and settled at room temperature for 10 minutes. After the signal was stable, the luminescence value was read on EnVision (Perkin Elmer 2104). Inhibition rate (%)=(1−Lum.sub.test drug/Lum.sub.negative control)×100. The negative control group was 0.667% DMSO. XL-fit software was used for calculation of IC50.

[0300] The compounds of the present invention were tested in the above-mentioned DU145 cell proliferation experiment. Compared with the non-deuterated compound Darolutamide, the compounds of the present invention have a comparable inhibitory effect on the proliferation of DU145 cells.

[0301] (4) Metabolic Stability Evaluation

[0302] Microsome experiment: human liver microsome: 0.5 mg/mL, Xenotech; rat liver microsome: 0.5 mg/mL, Xenotech; coenzyme (NADPH/NADH): 1 mM, Sigma Life Science; magnesium chloride: 5 mM, 100 mM phosphate buffer (pH 7.4).

[0303] Preparation of stock solution: a certain amount of the example compound powder and control compound powder were accurately weighed, and respectively dissolved to 5 mM with DMSO.

[0304] Preparation of phosphate buffer (100 mM, pH7.4): 150 mL of 0.5M potassium dihydrogen phosphate and 700 mL of 0.5M dipotassium hydrogen phosphate prepared beforehand were mixed, with pH of the mixture adjusted to 7.4 with 0.5M dipotassium hydrogen phosphate solution. Before use, the mixture was diluted by 5 folds with ultrapure water and magnesium chloride was added to obtain the phosphate buffer (100 mM), which contains 100 mM potassium phosphate, 3.3 mM magnesium chloride, and has pH 7.4.

[0305] NADPH regeneration system solution (containing 6.5 mM NADP, 16.5 mM G-6-P, 3 U/mL G-6-P D, 3.3 mM magnesium chloride) was prepared, and placed on wet ice before use.

[0306] Preparation of stop solution: a solution of 50 ng/mL propranolol hydrochloride and 200 ng/mL tolbutamide (internal standard) in acetonitrile. 25057.5 μL of phosphate buffer (pH 7.4) was taken into a 50 mL centrifuge tube, added with 812.5 μL of human liver microsome, and mixed well to obtain a liver microsome diluent with a protein concentration of 0.625 mg/mL. 25057.5 μL of phosphate buffer (pH 7.4) was taken into a 50 mL centrifuge tube, added with 812.5 μL of SD rat liver microsome, and mixed well to obtain a liver microsome diluent with a protein concentration of 0.625 mg/mL.

[0307] Incubation of the samples: the stock solutions of the corresponding compounds were respectively diluted to 0.25 mM with an aqueous solution containing 70% acetonitrile to obtain working solutions for later use. 398 μL of human liver microsome and rat liver microsome diluents were respectively taken and added to a 96-well incubation plate (N=2), added with 2 μL of 0.25 mM working solution respectively, and mixed well.

[0308] Determination of metabolic stability: 300 μL of pre-cooled stop solution was added to each well of a 96-well deep-well plate, and the plate was placed on ice as a stop plate. The 96-well incubation plate and the NADPH regeneration system were placed in a 37° C. water bath, shaken at 100 rpm, and pre-incubated for 5 minutes. 80 μL of incubation solution was taken out of each well of the incubation plate and added to the stop plate, mixed well, and supplemented with 20 μL of NADPH regeneration system solution to obtain the 0 min sample. Then 80 μL of NADPH regeneration system solution was added to each well of the incubation plate to initiate the reaction and start timing. The reaction concentration of the corresponding compound was 1 μM, and the protein concentration was 0.5 mg/mL. At 10, 30, and 90 minutes of reaction, 100 μL of the reaction solution was taken and added to the stop plate, and vortexed for 3 minutes to stop the reaction. The stop plate was centrifuged at 5000×g for 10 min at 4° C. 100 μL of supernatant was taken to a 96-well plate pre-added with 100 μL of distilled water, mixed well, and subjected to sample analysis by LC-MS/MS.

[0309] Data analysis: the peak areas of the corresponding compound and internal standard were detected through the LC-MS/MS system, and the ratio of the peak area of the compound to the internal standard was calculated. The natural logarithm of the remaining percentage of the compound was plotted against time to measure the slope and calculate t.sub.1/2 and CL.sub.int according to the following equations, where V/M is equal to 1/protein concentration.

[00001]$t_{1/2}=-\frac{0.693}{\mathrm{Slope}},{\mathrm{CL}}_{\mathrm{int}}=\frac{0.693}{t_{1/2}}.Math.\frac{V}{M},t_{1/2}\left(\min \right);C{L}_{\mathrm{int}}\left(\mathrm{.Math.L}\text{/}\min \text{/}\mathrm{mg}\right)$

[0310] The compounds of the present invention and the corresponding non-deuterated compounds were simultaneously tested for comparison and evaluated for the metabolic stability in human and rat liver microsomes. The non-deuterated compound Darolutamide was used as the control. In human and rat liver microsomal experiments, the compounds of the present invention significantly improved the metabolic stability, as compared with the non-deuterated compound Darolutamide. The results of representative example compounds are summarized in Table 1 below.

TABLE-US-00002 TABLE 1 RLM Example HLM CL Compounds T.sub.1/2 (min) CL (μL/min/mg) T.sub.1/2 (min) (μL/min/mg) Darolutamide 163.0 8.5 113.7 12.2 1 222.1 6.2 NA NA 2 152.8 9.1 NA NA 3 349.9 4.0 NA NA 4 162.2 8.5 NA NA 5 321.3 4.3 NA NA 6 217.0 6.4 NA NA 7 201.5 6.9 194.8 7.1 8 190.5 7.3 NA NA 9 171.2 8.1 NA NA 10 317.2 4.4 NA NA

[0311] (5) Rat Pharmacokinetics Experiment

[0312] 6 male Sprague-Dawley rats, 7-8 weeks old, weighing about 210 g, were divided into 2 groups, 3 rats in each group, and a single dose of the compound (10 mg/kg orally) was administered intravenously or orally to compare the pharmacokinetics.

[0313] The rats were fed with standard feed and given water, and fasting began 16 hours before the test. The drug was dissolved with PEG400 and dimethyl sulfoxide. Blood was collected from the orbit. The time points for blood collection were 0.083, 0.25, 0.5, 1, 2, 4, 6, 8, 12, and 24 hours after administration.

[0314] The rats were briefly anesthetized after inhaling ether, and a blood sample of 300 μL was collected from the orbit in a test tube. There was 30 μL of 1% heparin salt solution in the test tube. Before use, the test tube was dried overnight at 60° C. After the blood sample was collected at the last time point, the rats were anesthetized with ether and sacrificed.

[0315] Immediately after the blood sample was collected, the test tube was gently inverted at least 5 times to ensure thorough mixing and then placed on ice. The blood sample was centrifuged at 5000 rpm for 5 minutes at 4° C. to separate the plasma from the red blood cells. 100 μL of plasma was transferred with a pipette into a clean plastic centrifuge tube, labeled with the name of the compound and the time point. The plasma was stored at −80° C. before analysis. The concentration of the compound of the invention in the plasma was determined by LC-MS/MS. The pharmacokinetic parameters were calculated based on the blood drug concentrations of each animal at different time points.

[0316] Experiments showed that the compounds of the present invention had better pharmacokinetic properties in animals and thus had better pharmacodynamics and therapeutic effects.

[0317] The above content is a further detailed description of the present invention in conjunction with specific preferred embodiments, and it cannot be considered that the specific implementation of the present invention is just limited to the description. For those of ordinary skill in the technical field to which the present invention belongs, a number of simple deductions or substitutions can be made without departing from the concept of the present invention, which should be regarded as falling within the protection scope of the present invention.