A class of androgen receptor modulators, their preparation methods and applications

Abstract

This invention discloses a class of androgen receptor modulators, their preparation methods, and applications. The androgen receptor modulators are selected from compounds having the structural formula shown in Formula I. Experimental verification has shown that these compounds are all androgen receptor modulators, capable of significantly reducing the expression levels of androgen receptors in various cells and effectively inhibiting the proliferation of prostate cancer cells. They exhibit good safety profiles and have potential for further development.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, specifically relating to a class of androgen receptor modulators, their preparation methods, and applications. Background Technology

[0002] The androgen receptor (AR) belongs to the steroid receptor superfamily of nuclear receptors and is a type of transcriptional protein. An AR typically consists of four domains: the N-terminal activation domain (NTD), the DNA-binding domain (DBD), the hinge domain, and the ligand-binding domain (LBD).

[0003] Prostate cancer is a leading cause of cancer death in men. The androgen receptor (AR), a transcription factor belonging to the same family as p53, Myc, and NF-kappaB, has long been a major target in prostate cancer. Therefore, in addition to surgery and radiotherapy, androgen deprivation therapy is a first-line treatment for high-risk patients with localized prostate cancer, and many companies have developed drugs targeting AR. Currently available traditional small-molecule drugs targeting AR include Erleada (apalutamide), Xtandi (enzalutamide), and Nubeqa (darolutamide). However, these proteins have traditionally been considered challenging drug targets. Furthermore, resistance to traditional inhibitors has hindered the clinical application of these drugs.

[0004] Therefore, there is still a need in the field to develop compounds that are suitable as androgen receptor modulators and / or have better pharmacodynamics / pharmacokinetics. Summary of the Invention

[0005] The purpose of this invention is to provide a class of androgen receptor modulators, their preparation methods, and applications. This invention synthesized 20 compounds, all of which are androgen receptor modulators and have a significant effect in reducing androgen receptor expression.

[0006] To achieve the above-mentioned objectives, the present invention employs the following technical solution:

[0007] This invention provides a class of androgen receptor modulators selected from compounds having the structure shown in Formula I, or pharmaceutically acceptable salts thereof, or solvent compounds, or stereoisomers, or isotopic compounds:

[0008]

[0009]

[0010] in,

[0011] X is C or N;

[0012] Ring A and ring B are independently selected from 1, 2, 3, and 4 R groups, respectively.11 Replacement C 4-8 cycloalkyl, C 4-8 Heterocyclic alkyl, C 6-10 Aryl or C 5-10 Mixed aromatics;

[0013] L1 and L2 are independently selected from single bonds and C, respectively. 1-6 Alkyl, C 2-6 alkenyl, C 2-6 Alkyne group, O, S, NH, CO, SO, SO2; the C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 The alkynyl group contains one or two R groups. 12 Substituents;

[0014] The structural formula of the linking group is as follows, and it is selected from straight-chain or branched C1-C groups that are absent or have substituents. 12 Alkylene or C 4-8 Cycloalkyl; optionally, the carbon chain contains 1-4 heteroatoms selected from N, S, O, Si or P;

[0015]

[0016] Where, n1 = 1 - 7; n2 = 1 - 7; n3 = 1 - 7; n4 = 1 - 7;

[0017] R1, R2, R3, R4, R5, R6, R7, R8, R9, R 10 R 11 R 12 Each is independently selected from non-existent, H, halogen, and C. 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl, OH, NH2, CN, nitro, carboxyl, C 3-6 cycloalkyl; the C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl, OH, NH2, carboxyl, C 3-6 The cycloalkyl group contains one, two, or three atoms selected from halogens, C1, C2, and C3, respectively. 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Substituents of alkyl halogens, OH, NH2, CN, nitro, and carboxyl groups.

[0018] Furthermore, the androgen receptor modulator is specifically selected from compounds 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, and 20, with the following structural formulas:

[0019]

[0020]

[0021] Furthermore, the androgen receptor modulator is an androgen receptor degrader, which can reduce the expression level of androgen receptors in cells.

[0022] The present invention also provides a method for preparing the androgen receptor modulator, which includes the following steps:

[0023] The intermediate, DIPEA and DMF were reacted at high temperature under argon protection, and compounds 1, 4 and 10 were obtained by preparative liquid chromatography.

[0024] Alternatively, the intermediate can be dissolved in methanol, triethylamine can be added dropwise to adjust the pH, the reaction can be carried out at room temperature, sodium cyanoborohydride can be added under ice bath, and compounds 2, 3, 5, 6, 7, 8, 9, 11, 12, 13, 19, and 20 can be obtained by preparative liquid chromatography.

[0025] Alternatively, the intermediates, HATU, DIPEA, and DMF were reacted under argon protection, and compounds 14, 15, 16, 17, and 18 were obtained by preparative liquid chromatography.

[0026] Furthermore, the intermediate is a heat shock protein 90 inhibitor and an androgen receptor antagonist.

[0027] The present invention also provides a pharmaceutical composition comprising a compound having the structural formula shown in Formula I or a pharmaceutically acceptable salt thereof.

[0028] The present invention also provides the use of the androgen receptor modulator or the pharmaceutical composition of claim 6 in the preparation of antitumor drugs.

[0029] Furthermore, the tumor is prostate cancer.

[0030] Furthermore, the drug can inhibit the proliferation of prostate cancer cells.

[0031] Compared with the prior art, the present invention has the following advantages and beneficial effects:

[0032] This invention synthesizes 20 novel compounds from existing compounds. The synthesis method of these compounds is simple, and experimental verification shows that these compounds are all androgen receptor modulators, which can significantly reduce the expression level of androgen receptors in various cells and effectively inhibit the proliferation of prostate cancer cells. They have good safety and are worthy of further development. Detailed Implementation

[0033] The technical solution of the present invention will be further described in detail with reference to the following specific examples.

[0034] Unless otherwise specified, the experimental methods used in the following examples are all conventional methods, and the materials and reagents used can be purchased from biological or chemical reagent companies.

[0035] Example 1

[0036] 1. The synthetic route for intermediate S7 is as follows:

[0037]

[0038] Specifically:

[0039]

[0040] Compound S1 (2.5 g, 8.4 mmol) was added to a reaction flask, and HCl (20 mL, 80 mmol, 4 M 1,4-dioxane solution) was added dropwise at room temperature. The mixture was stirred for 1 h. The reaction solution was filtered, washed with 10 mL of ethyl acetate, and drained to dryness using an oil pump to give 1.96 g of white solid S2, with a yield of 100%. UPLC-MS was calculated for C8H8BrN[M+H]. + :197.98, 199.98,found:198.01, 200.02.

[0041]

[0042] S3 (863 mg, 2.3 mmol) and DCM (10 mL) were added to a reaction flask, purged three times with argon, and SOCl2 (230 mg, 2.8 mmol) and 1 drop of DMF were added. The mixture was stirred at room temperature for 2 h and concentrated to obtain acyl chloride. S2 (600 mg, 2.5 mmol), triethylamine (700 mg, 6.9 mmol), and DCM (10 mL) were added to a reaction flask, and acyl chloride was added dropwise. The mixture was stirred overnight under argon protection. 50 mL of water was added to the reaction solution, and the mixture was extracted with ethyl acetate (50 mL * 2), washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain 1.35 g of brown solid S4, with a yield of 100%. UPLC-MS calculated for C 32 H 30 BrNO3[M+H] + :556.14, 558.14,found:556.16, 558.15.

[0043]

[0044] S4 (1.2 g, 2.7 mmol), potassium (4-tert-butyloxycarbonylpiperazin-1-yl)methyltrifluoroborate (0.82 g, 2.7 mmol), XPhos (130 mg, 0.1 mmol), Cs₂CO₃ (2.6 g, 8.1 mmol), THF (20 mL), and water (2 mL) were added to a reaction flask. The mixture was purged with argon three times. Pd(OAc)₂ (30 mg, 0.135 mmol) was added, and the mixture was purged with argon three more times. The mixture was heated to reflux and stirred overnight. 200 mL of water was added to the reaction solution, and the mixture was extracted with ethyl acetate (100 mL * 2). The organic phase was washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was purified by column chromatography (DCM:EA = 3:1) to give 1.4 g of light brown solid S5, with a yield of 85%. UPLC-MS calculated for C 42 H 49 N3O5[M+H] + :676.37,found:676.39.

[0045]

[0046] S5 (530 mg) and acetic acid (10 mL) were added to a reaction flask, followed by 10% Pd / C (60 mg). The mixture was stirred overnight under hydrogen atmosphere, filtered, concentrated, and purified by column chromatography (DCM:MeOH = 25:1) to give 310 mg of pale yellow solid S6, with a yield of 83%. UPLC-MS calculated for C 28 H 37 N3O5[M+H] + :496.27,found:496.29.

[0047]

[0048] S6 (310 mg, 0.625 mmol) was added to a reaction flask, and HCl (1.6 mL, 6.4 mmol, 4 M 1,4-dioxane solution) was added dropwise at room temperature. The mixture was stirred for 1 h. 5 mL of methanol was added, the mixture was filtered, concentrated, and drained to dryness using an oil pump to give 300 mg of white solid S7, with a yield of 100%. UPLC-MS calculated for C6. 23 H 29 N3O3[M+H] + :396.22,found:396.25.

[0049] 2. The synthetic route for intermediate S13 is as follows:

[0050]

[0051] Specifically:

[0052]

[0053] NaH (50 mg, 1.23 mmol) was added to a 20 mL solution of DMF containing S9 (250 mg, 1.03 mmol) at 0 °C, and the mixture was stirred for 20 min. The starting material S8 (161 mg, 1.03 mmol) was added to the reaction system, and the mixture was allowed to react at room temperature for 4 h. TLC analysis showed that the reaction was complete. The mixture was extracted with ethyl acetate and water, washed with saturated brine, dried over anhydrous sodium sulfate, and then subjected to column chromatography (PE:EA = 8:1) to obtain 320 mg of a white solid S10, with a yield of 82%. UPLC-MS calculated for C 20 H 27 ClN2O3[M+H] + :378.90,found:378.92.

[0054]

[0055] Intermediate S10 (320 mg, 0.84 mmol) was dissolved in dichloromethane (20 mL), and 4 M hydrochloric acid / dioxane solution (2.5 mL) was added. The mixture was stirred at room temperature for 4 h. TLC showed that the reaction was complete. The solvent was removed to obtain a yellow solid, S11. UPLC-MS calculated for C 15 H 19 ClN2O[M+H] + :278.78,found:278.87.

[0056]

[0057] Intermediate S11 (75 mg, 0.24 mmol) and DIPEA (0.12 mL, 0.71 mmol) were dissolved in anhydrous dichloromethane (20 mL), and the mixture was stirred at room temperature for 20 min. Intermediate S12 (51 mg, 0.29 mmol) was then added, and the mixture was stirred overnight at room temperature under argon protection. After evaporating the solvent, column chromatography (PE:EA = 8:1) yielded 100 mg of a pale yellow solid, S13, in 99.37% yield. UPLC-MS calculated for C 20 H 20 Cl2N4O2[M+H] + :419.31,found:418.91.

[0058] 3. The synthetic route for intermediate S17 is as follows:

[0059]

[0060] Specifically:

[0061]

[0062] NaH (50 mg, 1.23 mmol) was added to a 20 mL solution of DMF containing S14 (222 mg, 1.03 mmol) at 0 °C, and the mixture was stirred for 20 min. The starting material S8 (161 mg, 1.03 mmol) was added to the reaction system, and the mixture was allowed to react at room temperature for 4 h. TLC analysis showed that the reaction was complete. The mixture was extracted with ethyl acetate and water, washed with saturated brine, dried over anhydrous sodium sulfate, and then subjected to column chromatography (PE:EA = 5:1) to obtain 280 mg of a white solid S15, with a yield of 77.48%. UPLC-MS calculated for C 18 H 23 ClN2O3[M+H] + :350.84,found:350.82.

[0063]

[0064] Intermediate S15 (280 mg, 0.80 mmol) was dissolved in dichloromethane (20 mL), and 4 M hydrochloric acid / dioxane solution (2.0 mL) was added. The mixture was stirred at room temperature for 4 h. TLC analysis showed that the reaction was complete. The solvent was evaporated to obtain a yellow solid, S16. UPLC-MS calculated for C 13 H 15 ClN2O[M+H] + :250.73,found:250.75.

[0065]

[0066] Intermediate S16 (500 mg, 1.7 mmol) and DIPEA (0.9 mL, 5.2 mmol) were dissolved in anhydrous dichloromethane (20 mL), and the mixture was stirred at room temperature for 20 min. Intermediate S12 (360 mg, 2.1 mmol) was then added, and the mixture was stirred overnight at room temperature under argon protection. After evaporating the solvent, column chromatography (PE:EA = 5:1) yielded 670 mg of a pale yellow solid, S17, in 100% yield. UPLC-MS calculated for C 18 H 16 Cl2N4O2[M+H] + :391.25,found:390.79.

[0067] 4. Synthesis of intermediate S19

[0068]

[0069] Intermediate S16 (50 mg, 0.17 mmol) and DIPEA (0.09 mL, 0.52 mmol) were dissolved in anhydrous dichloromethane (20 mL), and the mixture was stirred at room temperature for 20 min. Intermediate S18 (36 mg, 0.21 mmol) was then added, and the mixture was stirred overnight at room temperature under argon protection. After evaporating the solvent, column chromatography (PE:EA = 5:1) yielded 65 mg of a pale yellow solid, S19, in 99% yield. UPLC-MS calculated for C 18 H 16 Cl2N4O2[M+H] + :391.06,found:390.81.

[0070] 5. Synthesis of intermediate S21

[0071]

[0072] Intermediate S16 (36 mg, 0.21 mmol) and DIPEA (0.09 mL, 0.52 mmol) were dissolved in anhydrous dichloromethane (20 mL). The mixture was stirred at room temperature for 20 min. S20 (35 mg, 0.21 mmol) was then added, and the mixture was stirred overnight at room temperature under argon protection. After evaporating the solvent, column chromatography (PE:EA = 5:1) yielded 78 mg of a pale yellow solid, S21, in 99% yield. UPLC-MS calculated for C 18 H 16 Cl2N4O2[M+H] + :390.07,found:390.20.

[0073] 6. Synthesis of intermediate S25

[0074]

[0075] The synthetic route followed intermediate S17, but the starting material was replaced with 4-fluoro-2-(trifluoromethyl)benzonitrile (S22) instead of 2-chloro-4-fluorobenzonitrile (S8), yielding a pale yellow solid S25 (60 mg). UPLC-MS was calculated for C... 19 H 16 ClF3N4O2[M+H] + :425.10,found:425.24.

[0076] 7. The synthetic route for intermediate S28 is as follows:

[0077]

[0078] Specifically:

[0079]

[0080] Monomethyl terephthalate S26 (80 mg, 0.28 mmol), HATU (158 mg, 0.42 mmol), and DIPEA (0.24 mL, 1.38 mmol) were dissolved in anhydrous DMF. The mixture was stirred for 15 min under argon protection. A DMF solution of S16 (50 mg, 0.28 mmol) was added, and the mixture was stirred overnight at room temperature. The mixture was extracted with water and ethyl acetate. The organic phase was washed successively with saturated ammonium chloride, saturated sodium bicarbonate, and saturated brine. After drying with anhydrous magnesium sulfate, column chromatography (PE:EA = 2:1) yielded 77 mg of solid S27, in 67% yield. UPLC-MS calculated for C 22 H 21 ClN2O4[M+H] + :413.12,found:413.62.

[0081]

[0082] Intermediate S27 (70 mg, 0.17 mmol) was dissolved in tetrahydrofuran solution, and lithium hydroxide (72 mg, 1.7 mmol) and a small amount of water were added. The mixture was stirred overnight at room temperature. TLC was used to confirm the complete reaction of the starting material. The organic phase was evaporated to dryness, and water and ethyl acetate were added for extraction (the organic phase was discarded). The pH of the aqueous phase was adjusted to 2-3 with hydrochloric acid, and the mixture was extracted three times with ethyl acetate. The combined organic phases were washed with saturated brine and dried over anhydrous magnesium sulfate to obtain 60 mg of crude intermediate S28.

[0083] 8. The synthetic route for intermediate S32 is as follows:

[0084]

[0085] Specifically:

[0086]

[0087] 2-Chloro-4-fluorobenzonitrile (300 mg, 1.93 mmol) was dissolved in dimethylformamide (20 mL), and DIEA (1.3 g, 10 mmol) and trans-(4-aminocyclohexyl)carbamate tert-butyl ester S29 (455.6 mg, 2.12 mmol) were added. The mixture was heated to 90 °C and reacted for 18 h. After cooling to room temperature, the mixture was diluted with 100 mL of water, extracted with ethyl acetate (30 mL * 3), washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was purified by column chromatography (PE:EA = 1:1) to give 250 mg of white solid S30, yield 37%. UPLC-MS calculated for C 18 H 24 ClN3O2[M+H] + :350.16,found:350.22.

[0088] The remaining synthetic route followed intermediate S17, yielding 260 mg of a pale yellow solid, S32. UPLC-MS was calculated for C... 18 H 17 ClN2O5[M+H] + :390.08,found:390.12.

[0089] 9. The synthetic route for intermediate S36 is as follows:

[0090]

[0091] Specifically:

[0092]

[0093] 4-Fluoro-2-(trifluoromethyl)benzonitrile S22 (500 mg, 2.64 mmol) and 1-Boc-piperazine S33 (800 mg, 4.29 mmol) were dissolved in dimethylformamide (20 mL) and reacted at 80 °C for 3 h. After cooling to room temperature, water (200 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (30 mL x 3). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was purified by column chromatography (PE:EA = 5:1) to give 770 mg of white solid S34, yield 82%. UPLC-MS calculated for C 17 H 20 F3N3O2[M+H]+:356.15,found:356.30.

[0094] The remaining synthetic route followed intermediate S17, yielding 260 mg of a pale yellow solid S38. UPLC-MS was calculated for C...17 H 13 ClN3O5[M+H] + :396.08,found:396.77.

[0095] 10. The synthetic route for intermediate S45 is as follows:

[0096]

[0097] Specifically:

[0098]

[0099] Starting materials S37 (3 g, 13.8 mmol), S38 (5.8 g, 20.8 mmol), Pd(dppf)Cl2 (1 g, 1.3 mmol), and K2CO3 (6.0 g, 41.6 mmol) were dissolved in dioxane (50 ml) and H2O (5 ml). The mixture was stirred overnight at 85 °C under argon protection. The mixture was extracted with water and ethyl acetate. The organic phase was washed successively with saturated ammonium chloride and saturated brine, dried over anhydrous magnesium sulfate, and subjected to column chromatography (PE:EA = 1:1) to give solid S39 3.0 g, yield 75%. UPLC-MS calculated for C 15 H 14 ClN3O[M+H] + :288.02,found:288.15.

[0100]

[0101] Intermediate S39 (1 g, 3.48 mmol) was dissolved in MeOH (10 ml). A 4 M hydrochloric acid / dioxane solution (2 ml) was added dropwise to the system at 0 °C, and the mixture was stirred at room temperature for 4 h. The organic phase was concentrated to give 800 mg of solid S40.

[0102]

[0103] Intermediates S40 (500 mg, 2.46 mmol), S41 (470 mg, 21.61 mmol), and triphenylphosphine were dissolved in dry THF under nitrogen protection. DBAD (0.96 mL, 4.91 mmol) was added dropwise under ice bath conditions, and the mixture was stirred overnight (18 h) at room temperature. Column chromatography yielded 487 mg of solid S42, 55% yield. UPLC-MS calculated for C1. 18 H 21 ClN4O2[M+H] + :361.14,found:361.28.

[0104]

[0105] Intermediate S42 (450 mg, 1.25 mmol) was dissolved in MeOH (10 mL). A 4 M hydrochloric acid / dioxane solution (1 mL) was added dropwise to the system at 0 °C, and the mixture was stirred at room temperature for 4 h. The organic phase was concentrated to obtain solid S43360 mg.

[0106]

[0107] S44 (132 mg, 1.18 mmol) and DIPEA (0.3 mL, 1.77 mmol) were dissolved in 10 mL of dry DCM. HOBT (240 mg, 1.77 mmol), EDCI (340 mg, 1.77 mmol), and S43 (350 mg, 1.18 mmol) were added at room temperature. The reaction mixture was stirred overnight at room temperature. 40 mL of DCM was added, and the mixture was washed with (15 mL x 3) water, dried over Na2SO4, filtered, and evaporated to dryness. Column chromatography (DCM:MeOH = 50:1) yielded 153 mg of solid S45, 37% yield. UPLC-MS calculated for C 17 H 15 ClN6O[M+H] + :355.10,found:355.80.

[0108] Example 2

[0109] 1. Synthesis of Compound 1

[0110]

[0111] Intermediates S7 (70 mg, 0.16 mmol), S13 (67 mg, 0.16 mmol), DIPEA (82 mg, 0.64 mmol), and DMF (2 mL) were added to a reaction flask, purged three times with argon, stirred overnight at 85 °C, concentrated to obtain a crude product, and purified by reverse phase preparation to obtain 25 mg of compound 1 solid, yield 20%.

[0112] UPLC-MS calculated for C 43 H 48 ClN7O5[M+H] + :778.34,found:779.35.

[0113] 1H NMR (400MHz, DMSO-d6) δ10.08(s,1H),9.64(s,1H),8.06(d,J=3.4Hz,1H),8.03(d,J=0.9Hz,1H),7.59(d,J=0.6Hz,1H),7.46(dd,J=7.5,5.8 Hz,2H),7.30(h,J=1.1Hz,1H),7.25(dt,J=7.5,1.1Hz,1H),7.17(d,J=1.4Hz,1H),7.15(dq,J=7.5,1.1Hz,1H),7.03(dd,J=7.5,1.5Hz,1H),5 .37(ddq,J=4.4,3.1,1.5Hz,1H),4.55–4.52(m,2H),4.49–4.47(m,2H),3.81(dddt,J=11.8,4.1,2.9,1.4Hz,1H),3.63(t,J=1.0Hz,2H),3.57 (t,J=7.1Hz,4H),3.17(dtd,J=13.6,6.8,0.7Hz,1H),2.77–2.62(m,4H),1.22(d,J=6.7Hz,6H),1.10(t,J=1.4Hz,6H),1.05(t,J=1.4Hz,6H).

[0114] 2. The synthetic route for compound 2 is as follows:

[0115]

[0116] Specifically:

[0117]

[0118] S11 (879 mg, 2.80 mmol) was dissolved in dimethylformamide (20 mL), followed by the addition of diisopropylethylamine (1.1 g, 8.52 mmol), HATU (1.3 g, 3.42 mmol), and S44 (345 mg, 2.80 mmol). The reaction was carried out at room temperature for 18 h. Then, 1 M sodium hydroxide solution (10 mL) was added to the reaction mixture, and the reaction was carried out at room temperature for 2 h. A saturated ammonium chloride solution (200 mL) was added, and the mixture was extracted with ethyl acetate (50 mL x 3). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was evaporated to dryness and purified by column chromatography (DCM = 10:1) to give solid S46 917 mg, yield 88%. UPLC-MS calculated for C 19 H 21 ClN4O2[M+H]+:373.14,found:373.85.

[0119]

[0120] S46 (390 mg, 1.05 mmol) was dissolved in dimethylformamide (10 mL), followed by the addition of potassium carbonate (290 mg, 2.10 mg), potassium iodide (35 mg, 0.21 mol), and S47 (263 mg, 1.56 mmol). The mixture was heated to 90 °C and reacted for 48 h. After cooling to room temperature, water (100 mL) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (20 mL x 3). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was purified by column chromatography (PE:EA = 1:10) to obtain 430 mg of white solid S48, with a yield of 89%. UPLC-MS calculated for C 23 H 29 ClN4O4[M+H]+:461.19,found:461.96.

[0121]

[0122] S48 (129 mg, 0.28 mmol) was dissolved in tetrahydrofuran (20 mL), and 6 M hydrochloric acid solution (10 mL) was added. The mixture was heated to 40 °C and reacted for 18 h. After cooling to room temperature, water (100 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (30 mL x 3). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was evaporated to dryness to obtain a yellow solid crude product S49 97 mg, yield 84%. UPLC-MS calculated for C 21 H 23 ClN4O3[M+H]+:415.14,found:415.89.

[0123]

[0124] S49 (85 mg, 0.21 mmol) and S7 (88 mg, 0.21 mmol) were dissolved together in methanol (5 mL). Triethylamine was added dropwise to adjust the pH to approximately 8. The mixture was stirred at room temperature for 30 min. Sodium cyanoborohydride (10 mg, 0.26 mmol) was added under ice bath conditions, and the mixture was stirred at room temperature overnight. 51 mg of compound 2 was obtained as a white powder by preparative liquid chromatography, with a yield of 31%.

[0125] UPLC-MS calculated for C 44 H 52 ClN7O5[M+H]+:794.37,found:795.39.

[0126] 1H NMR (400MHz, DMSO-d6) δ10.08(s,1H),9.64(s,1H),8.06(d,J=1.4Hz,1H),7.84(dd,J=6.3,4.8Hz,2H),7.77(d,J=7.5Hz,1H),7.59(d ,J=0.6Hz,1H),7.29(h,J=1.0Hz,1H),7.25(dt,J=7.5,1.1Hz,1H),7.19–7.11(m,2H),7.01(dd,J=7.4,1.4Hz,1H),6.55(s,1H),5.37 (pq,J=3.1,1.5Hz,1H),4.56–4.51(m,2H),4.51–4.46(m,2H),4.31(t,J=7.1Hz,2H),3.92–3.82(m,1H),3.63(t,J=1.0Hz,2H),3.17( dtd,J=13.6,6.8,0.7Hz,1H),2.93(t,J=7.1Hz,2H),2.53(s,8H),1.22(d,J=6.7Hz,6H),1.11(t,J=1.4Hz,6H),1.06(t,J=1.4Hz,6H).

[0127] 3. The synthetic route for compound 3 is as follows:

[0128]

[0129] Specifically:

[0130]

[0131] Intermediate S13 (100 mg, 0.24 mmol) was dissolved together with 4-hydroxymethylpiperidine S50 (33 mg, 0.29 mmol) and K2CO3 (64 mg, 0.48 mmol) in DMF (20 mL), and the mixture was stirred at 80 °C for 2 h. TLC analysis showed complete reaction of the starting material. After evaporation of the solvent, column chromatography (DCM:MeOH = 25:1) yielded 100 mg of a yellow solid S51, with a yield of 84%. UPLC-MS calculated for C 26 H 32 ClN5O3[M+H] + :498.22,found:497.97.

[0132]

[0133] Intermediate S51 (70 mg, 0.14 mmol) was dissolved in dichloromethane (20 mL), and DMP (102 mg, 0.24 mmol) was slowly added to the reaction flask. The mixture was stirred at room temperature for 2 h. TLC analysis showed that the reaction proceeded completely. The organic phase was washed successively with saturated sodium bicarbonate and saturated brine, dried over anhydrous sodium sulfate, and after evaporation of the solvent, column chromatography (DCM:MeOH = 50:1) yielded a pale yellow solid, 260 mg, with a yield of 86%. UPLC-MS calculated for C 26 H 30 ClN5O3[M+H] + :496.20,found:495.91.

[0134]

[0135] S52 (50 mg, 0.10 mmol) and S7 (43 mg, 0.10 mmol) were dissolved together in methanol (5 mL). Triethylamine was added dropwise to adjust the pH to approximately 8. The mixture was stirred at room temperature for 30 min. Sodium cyanoborohydride (8 mg, 0.2 mmol) was added under ice bath conditions, and the mixture was stirred at room temperature overnight. 35 mg of compound 3 was obtained as a white powder by preparative liquid chromatography, with a yield of 40%.

[0136] UPLC-MS calculated for C 49 H 59 ClN8O5[M+H]+:875.43,found:875.62.

[0137] 1H NMR (400MHz, DMSO-d6) δ10.08(s,1H),9.64(s,1H),8.08–8.01(m,2H),7.59(d,J=0.6Hz,1H),7.46(dd,J=7.5,5.8Hz,2 H),7.30–7.21(m,2H),7.19–7.11(m,2H),7.03(dd,J=7.5,1.5Hz,1H),6.55(s,1H),5.37(pq,J=3.1,1.5Hz,1H),4.53(d ,J=0.8Hz,2H),4.48(d,J=1.1Hz,2H),3.90–3.79(m,1H),3.72–3.60(m,4H),3.41–3.29(m,2H),3.17(dtd,J=13.6,6.8, 0.7Hz, 1H), 2.66–2.43 (m, 10H), 1.95–1.83 (m, 5H), 1.22 (d, J = 6.7Hz, 6H), 1.10 (t, J = 1.5Hz, 6H), 1.05 (t, J = 1.5Hz, 6H).

[0138] 4. Synthesis of Compound 4

[0139]

[0140] Following the synthetic route of compound 1, we obtained 40 mg of white solid compound 4.

[0141] UPLC-MS calculated for C 41 H 44 ClN7O5[M+H] + :750.31,found:750.36.

[0142] 1H NMR (400MHz, DMSO-d6) δ10.08(s,1H),9.65(s,1H),8.62(d,J=8.2Hz,1H),7.84(dd,J=11.4,9.1Hz,2H),7.39(d,J=2. 4Hz,1H),7.33(d,J=9.5Hz,2H),7.25(s,2H),7.13(dd,J=8.8,2.5Hz,1H),7.04(s,1H),6.40(s,1H),4.77(s,4H),4.52 (tt,J=9.9,4.1Hz,1H),3.94–3.78(m,1H),3.69(d,J=6.2Hz,4H),3.52(s,3H),3.10(h,J=6.8Hz,1H),2.47(d,J=5.4H z,2H),2.15–2.04(m,2H),1.94–1.81(m,2H),1.72–1.57(m,2H),1.50(dt,J=14.1,10.8Hz,2H),1.13(d,J=7.0Hz,6H).

[0143] 5. Synthesis of Compound 5

[0144]

[0145] Following the synthetic route of compound 3, 5.42 mg of a white solid compound was obtained. UPLC-MS was calculated for C... 47 H 55 ClN8O5[M+H] + :847.40,found:847.48. 1H NMR (400MHz, DMSO-d6) δ10.08(s,1H),9.64(s,1H),8.59(d,J=8.2Hz,1H),7.84(d,J=8.8Hz,1H),7.78(d,J=9.5Hz,1H),7.38( d,J=2.5Hz,1H),7.30(d,J=9.7Hz,2H),7.19(s,2H),7.12(dd,J=8.8,2.4Hz,1H),7.03(s,1H),6.39(s,1H),4.75(s,4H),4.58 –4.37(m,3H),3.83(ddd,J=11.1,7.6,3.7Hz,1H),3.27(s,2H)3.07(q,J=6.9Hz,1H),2.96(d,J=24.8Hz,2H),2.34(s,7H),2.0 9(t,J=8.5Hz,5H),1.97–1.82(m,3H),1.78(s,2H),1.75(s,1H),1.69–1.56(m,2H),1.56–1.39(m,2H),1.12(d,J=6.9Hz,6H).

[0146] 6. Synthesis of Compound 6

[0147]

[0148] Following the synthetic route of compound 3, 6.37 mg of a white solid compound was obtained.

[0149] UPLC-MS calculated for C 46 H 55 F3N8O5[M+H] + :881.42,found:881.01.

[0150] 1H NMR (500MHz, DMSO-d6) δ10.08(s,1H),9.64(s,1H),7.92(d,J=7.5Hz,1H),7.78(d,J=7.4Hz,1H),7.59(d,J=0.6Hz,1H),7.51(d,J= 7.5Hz,1H),7.30–7.22(m,2H),7.19(d,J=1.5Hz,1H),7.15(dq,J=7.6,1.2Hz,1H),7.01(dd,J=7.5,1.5Hz,1H),6.55(s,1H),4.75(s ,4H),4.58–4.37(m,3H),3.83(ddd,J=11.1,7.6,3.7Hz,1H),3.27(s,2H)3.07(q,J=6.9Hz,1H),2.96(d,J=24.8Hz,2H),2.34(s,7H ), 2.09(t,J=8.5Hz,5H),1.97–1.82(m,3H),1.78(s,2H),1.75(s,1H),1.69–1.56(m,2H),1.56–1.39(m,2H),1.12(d,J=6.9Hz,6H).

[0151] 7. Synthesis of Compound 7

[0152]

[0153] Following the synthetic route of compound 3, 7.28 mg of a white solid compound was obtained.

[0154] UPLC-MS calculated for C 46 H 53 ClN8O5[M+H] + :833.38,found:833.43.

[0155] 1H NMR (400MHz, DMSO-d6) δ10.08(s,1H),9.64(s,1H),8.12–8.05(m,1H),8.03(s,1H),7.77(d,J=7.4Hz,1H),7.62–7.56( m,2H),7.30–7.21(m,2H),7.20–7.11(m,2H),6.99(dd,J=7.5,1.5Hz,1H),6.55(s,1H),4.56–4.51(m,2H),4.48(d,J=1 .1Hz,2H),4.20–4.09(m,1H),3.81(dt,J=12.5,7.1Hz,2H),3.77–3.67(m,1H),3.67–3.59(m,4H),3.17(dtd,J=13.6,6 .8,0.7Hz,1H),2.69–2.55(m,8H),2.41(p,J=7.0Hz,1H),1.98–1.71(m,10H),1.69–1.56(m,2H),1.12(d,J=6.7Hz,6H).

[0156] 8. Synthesis of Compound 8

[0157]

[0158] Following the synthetic route of compound 3, 8.47 mg of a white solid compound was obtained.

[0159] UPLC-MS calculated for C 48 H 57 ClN8O5[M+H] + :861.41,found:861.49.

[0160] 1H NMR (400MHz, DMSO-d6) δ10.08(s,1H),9.64(s,1H),8.12–8.01(m,2H),7.77(d,J=7.4Hz,1H),7.59(d,J=0.6Hz,1H),7.52(d,J= 7.5Hz,1H),7.30–7.21(m,2H),7.20–7.11(m,2H),6.99(dd,J=7.5,1.5Hz,1H),6.55(s,1H),4.53(d,J=0.8Hz,2H),4.51–4.46(m ,2H),4.20–4.09(m,1H),3.74(ddt,J=23.9,12.3,7.0Hz,3H),3.63(t,J=1.0Hz,2H),3.32(dt,J=12.4,7.1Hz,2H),3.17(dtd,J= 13.6, 6.8, 0.7Hz, 1H), 2.65–2.48 (m, 10H), 1.98–1.73 (m, 10H), 1.69–1.52 (m, 3H), 1.46 (q, J = 7.2Hz, 2H), 1.13 (d, J = 6.7Hz, 6H).

[0161] 9. Synthesis of Compound 9

[0162]

[0163] Following the synthetic route of compound 3, 9.31 mg of a white solid compound was obtained.

[0164] UPLC-MS calculated for C 48 H 56 ClN7O5[M+H] + :846.40,found:846.47.

[0165] 1H NMR (400MHz, DMSO-d6) δ10.08(s,1H),9.64(s,1H),8.76(d,J=1.5Hz,1H),8.03(d,J=11.5Hz,1H),7.92(dd,J=7.5,1.5Hz,1H),7 .77(d,J=7.4Hz,1H),7.59(d,J=0.6Hz,1H),7.30–7.21(m,2H),7.20–7.11(m,2H),7.05–6.96(m,2H),6.55(s,1H),4.75(s,4H), 4.58–4.37(m,3H),3.83(ddd,J=11.1,7.6,3.7Hz,1H),3.27(s,2H)3.07(q,J=6.9Hz,1H),2.96(d,J=24.8Hz,2H),2.34(s,7H),2 .09(t,J=8.5Hz,5H),1.97–1.82(m,3H),1.78(s,2H),1.75(s,1H),1.69–1.56(m,2H),1.56–1.39(m,2H),1.12(d,J=6.9Hz,6H).

[0166] 10. The synthetic route of compound 10 is as follows:

[0167]

[0168] Specifically:

[0169]

[0170] Intermediate S17 (80 mg, 0.21 mmol) was dissolved together with starting material S65 (35 mg, 0.25 mmol) and K2CO3 (69 mg, 0.50 mmol) in DMF (20 mL), and the mixture was stirred at 80 °C for 2 h. TLC analysis showed complete reaction of the starting material. After evaporation of the solvent, column chromatography (DCM:MeOH = 25:1) yielded 82 mg of solid S66, with a yield of 79%. UPLC-MS calculated for C 25 H 28 ClN5O4[M+H] + :498.18,found:497.98.

[0171]

[0172] Intermediate S66 (80 mg, 0.16 mmol) was dissolved in tetrahydrofuran solution, and lithium hydroxide (68 mg, 1.6 mmol) and a small amount of water were added. The mixture was stirred overnight at room temperature. TLC analysis showed that the reaction proceeded completely. The organic phase was evaporated to dryness, and extracted with water and ethyl acetate (the organic phase was discarded). The aqueous phase was adjusted to pH 2-3 with hydrochloric acid, and extracted three times with ethyl acetate. The combined organic phases were washed with saturated brine and dried over anhydrous magnesium sulfate to obtain 55 mg of crude intermediate S67. UPLC-MS calculated for C 24 H 26 ClN5O4[M+H] + :484.17,found:483.95.

[0173]

[0174] Intermediate S67 (50 mg, 0.10 mmol), HATU (57 mg, 0.15 mmol), and DIPEA (0.05 mL, 0.30 mmol) were dissolved in anhydrous DMF. The mixture was stirred for 15 min under argon protection. Then, a DMF solution of S7 (47 mg, 0.11 mmol) was added, and the mixture was stirred overnight at room temperature. The solution was prepared by liquid chromatography to give 48 mg of compound 10 as a white solid, in 56% yield.

[0175] UPLC-MS calculated for C 47 H 53 ClN8O6[M+H] + :861.38,found:861.44.

[0176] 1H NMR (400MHz, DMSO-d6) δ10.08(s,1H),9.64(s,1H),8.12–8.05(m,1H),8.03(s,1H),7.77(d,J=7.4Hz,1H),7.59(d,J=0.6Hz ,1H),7.46(d,J=7.5Hz,1H),7.30–7.21(m,2H),7.20–7.11(m,2H),6.99(dd,J=7.5,1.5Hz,1H),6.55(s,1H),4.56–4.51(m, 2H),4.51–4.46(m,2H),4.20–4.09(m,1H),3.78–3.59(m,5H),3.50(d,J=1.6Hz,2H),3.49–3.41(m,4H),3.17(dtd,J=13.6, 6.8, 0.7Hz, 1H), 2.55 (p, J = 7.0Hz, 1H), 2.47 (t, J = 7.1Hz, 4H), 2.04–1.73 (m, 10H), 1.69–1.56 (m, 2H), 1.11 (d, J = 6.7Hz, 6H).

[0177] 11. Synthesis of Compound 11

[0178]

[0179] Following the synthetic route of compound 3, 11.29 mg of a white solid compound was obtained.

[0180] UPLC-MS calculated for C 45 H 51 ClN8O5[M+H] + :819.37,found:819.41.

[0181] 1H NMR (400MHz, DMSO-d6) δ10.08(s,1H),9.64(s,1H),8.12–8.01(m,2H),7.77(d,J=7.4Hz,1H),7.61–7.53(m,2H),7.29(h,J= 1.0Hz,1H),7.25(dt,J=7.5,1.1Hz,1H),7.20–7.11(m,2H),6.99(dd,J=7.5,1.5Hz,1H),6.55(s,1H),4.53(d,J=0.9Hz,2H) ,4.51–4.46(m,2H),4.20–4.09(m,1H),3.81(dd,J=12.4,6.9Hz,2H),3.78–3.61(m,5H),3.17(dtd,J=13.6,6.8,0.7Hz,1H) ,2.53(d,J=1.5Hz,8H),2.45–2.39(m,2H),2.39–2.25(m,1H),1.98–1.73(m,6H),1.69–1.56(m,2H),1.13(d,J=6.7Hz,6H).

[0182] 12. Synthesis of Compound 12

[0183]

[0184] Following the synthetic route of compound 3, 12.41 mg of a white solid compound was obtained.

[0185] UPLC-MS calculated for C 47 H 55 ClN8O5[M+H] + :847.40,found:847.46.

[0186] 1H NMR (400MHz, DMSO-d6) δ10.08(s,1H),9.64(s,1H),8.79(s,2H),8.15(d,J=11.5Hz,1H),7.77(d,J=7.4Hz,1H),7.59(d,J =0.6Hz,1H),7.30–7.21(m,2H),7.20–7.11(m,2H),6.99(dd,J=7.5,1.5Hz,1H),6.55(s,1H),4.75(s,4H),4.58–4.37(m, 3H),3.83(ddd,J=11.1,7.6,3.7Hz,1H),3.27(s,2H)3.07(q,J=6.9Hz,1H),2.96(d,J=24.8Hz,2H),2.34(s,7H),2.09(t, J=8.5Hz,5H),1.97–1.82(m,3H),1.78(s,2H),1.75(s,1H),1.69–1.56(m,2H),1.56–1.39(m,2H),1.12(d,J=6.9Hz,6H).

[0187] 13. Synthesis of Compound 13

[0188]

[0189] Following the synthetic route of compound 3, 13.87 mg of a white solid compound was obtained.

[0190] UPLC-MS calculated for C 47 H 56 ClN9O4[M+H] + :847.40,found:847.46.

[0191] 1H NMR (400MHz, DMSO-d6) δ10.08(s,1H),9.64(s,1H),8.79(s,2H),8.15(d,J=11.5Hz,1H),7.77(d,J=7.4Hz,1H),7.59(d,J =0.6Hz,1H),7.30–7.21(m,2H),7.20–7.11(m,2H),6.99(dd,J=7.5,1.5Hz,1H),6.55(s,1H),4.75(s,4H),4.58–4.37(m, 3H),3.83(ddd,J=11.1,7.6,3.7Hz,1H),3.27(s,2H)3.07(q,J=6.9Hz,1H),2.96(d,J=24.8Hz,2H),2.34(s,7H),2.09(t, J=8.5Hz,5H),1.97–1.82(m,3H),1.78(s,2H),1.75(s,1H),1.69–1.56(m,2H),1.56–1.39(m,2H),1.12(d,J=6.9Hz,6H).

[0192] 14. The synthetic route for compound 14 is as follows:

[0193]

[0194] Specifically:

[0195]

[0196] Starting materials S75 (100 mg, 0.46 mmol), HATU (263 mg, 0.69 mmol), and DIPEA (0.24 mL, 1.38 mmol) were dissolved in anhydrous DMF. The mixture was stirred for 15 min under argon protection. A DMF solution of S7 (198 mg, 0.46 mmol) was added, and the mixture was stirred overnight at room temperature. The mixture was extracted with water and ethyl acetate. The organic phase was washed successively with saturated ammonium chloride, saturated sodium bicarbonate, and saturated brine. After drying with anhydrous magnesium sulfate, column chromatography (DCM:MeOH = 15 / 1) yielded 123 mg of solid S76, with a yield of 45%. UPLC-MS calculated for C 33 H 46 N4O6[M+H] + :595.34,found:595.75.

[0197]

[0198] Intermediate S76 (100 mg, 0.17 mmol) was dissolved in MeOH (10 mL), and 4 M hydrochloric acid / dioxane solution (1 mL) was added dropwise to the system at 0 °C. The mixture was stirred at room temperature for 4 h. The organic phase was concentrated to obtain solid S7787 mg.

[0199]

[0200] Starting materials S28 (64 mg, 0.16 mmol), HATU (91 mg, 0.24 mmol), and DIPEA (0.08 mL, 0.48 mmol) were dissolved in anhydrous DMF. The mixture was stirred for 15 min under argon protection. Then, a DMF solution of S77 (85 mg, 0.16 mmol) was added, and the mixture was stirred overnight at room temperature. A white solid compound of 14.53 mg was obtained by liquid chromatography, with a yield of 38%.

[0201] UPLC-MS calculated for C 49 H 55 ClN6O7[M+H] + :875.38,found:875.46.

[0202] 1 H NMR (500MHz, DMSO-d6) δ10.08(s,1H),9.64(s,1H),8.01(d,J=11.5Hz,1H),7.94(t,J=6.7Hz,1H),7.89(s,4H),7.77(d,J=7.4H z,1H),7.59(d,J=0.6Hz,1H),7.30–7.22(m,2H),7.19–7.12(m,2H),6.99(dd,J=7.7,1.5Hz,1H),6.55(s,1H),4.53(d,J=1.1Hz, 2H),4.48(d,J=1.1Hz,2H),4.18–4.09(m,1H),3.65–3.54(m,3H),3.46(t,J=7.1Hz,4H),3.31(q,J=6.9Hz,2H),3.22–3.11(m,1H ), 2.47(t,J=7.1Hz,4H),2.33(t,J=6.9Hz,2H),1.97–1.87(m,2H),1.87–1.75(m,4H),1.72–1.52(m,6H),1.10(d,J=6.7Hz,6H).

[0203] 15. Synthesis of Compound 15

[0204]

[0205] Following the synthetic route of compound 14, we obtained 41 mg of white solid compound 15.

[0206] UPLC-MS calculated for C 51 H 59 ClN6O7[M+H] + :903.41,found:903.52.

[0207] 1 H NMR (500MHz, DMSO-d6) δ10.08(s,1H),9.64(s,1H),8.01(d,J=11.5Hz,1H),7.94(t,J=6.8Hz,1H),7.89(s,4H),7.77(d,J=7.4H z,1H),7.59(d,J=0.6Hz,1H),7.30–7.22(m,2H),7.19–7.12(m,2H),7.00(dd,J=7.5,1.5Hz,1H),6.55(s,1H),4.53(d,J=1.0Hz, 2H),4.48(d,J=1.1Hz,2H),4.18–4.09(m,1H),3.65–3.54(m,3H),3.46(t,J=7.1Hz,4H),3.28(q,J=7.0Hz,2H),3.22–3.11(m,1H ), 2.47(t,J=7.1Hz,4H),2.33(t,J=7.1Hz,2H),1.95–1.75(m,6H),1.67–1.53(m,6H),1.38–1.27(m,4H),1.11(d,J=6.7Hz,6H).

[0208] 16. Synthesis of Compound 16

[0209]

[0210] Following the synthetic route of compound 14, we obtained 35 mg of white solid compound 16.

[0211] UPLC-MS calculated for C 50 H 57 ClN6O7[M+H] + :889.40,found:889.49.

[0212] 1H NMR (500MHz, DMSO-d6) δ10.08(s,1H),9.64(s,1H),8.01(d,J=11.5Hz,1H),7.94(t,J=6.8Hz,1H),7.89(s,4H),7.77(d,J=7.4Hz,1H ),7.59(d,J=0.6Hz,1H),7.30–7.22(m,2H),7.19–7.12(m,2H),7.00(dd,J=7.5,1.5Hz,1H),6.55(s,1H),4.53(d,J=1.0Hz,2H),4.48 (d,J=1.1Hz,2H),4.18–4.09(m,1H),3.65–3.54(m,3H),3.46(t,J=7.1Hz,4H),3.26(q,J=7.0Hz,2H),3.22–3.11(m,1H),2.47(t,J=7 .1Hz,4H),2.35(t,J=7.1Hz,2H),1.97–1.87(m,2H),1.87–1.75(m,4H),1.72–1.52(m,6H),1.44–1.35(m,2H),1.12(d,J=6.7Hz,6H).

[0213] 17. Synthesis of Compound 17

[0214]

[0215] Following the synthetic route of compound 14, we obtained 17 52 mg of white solid compound.

[0216] UPLC-MS calculated for C 52 H 61 ClN6O7[M+H] + :917.43,found:917.54.

[0217] 1H NMR (500MHz, DMSO-d6) δ10.08(s,1H),9.64(s,1H),8.01(d,J=11.5Hz,1H),7.94(t,J=6.7Hz,1H),7.89(s,4H),7.77(d,J=7.4Hz,1H ),7.59(d,J=0.6Hz,1H),7.30–7.22(m,2H),7.19–7.12(m,2H),7.00(dd,J=7.5,1.5Hz,1H),6.55(s,1H),4.53(d,J=1.1Hz,2H),4.48 (d,J=1.1Hz,2H),4.18–4.09(m,1H),3.65–3.54(m,3H),3.46(t,J=7.1Hz,4H),3.28(q,J=7.0Hz,2H),3.22–3.11(m,1H),2.47(t,J=7 .1Hz,4H),2.34(t,J=7.1Hz,2H),1.97–1.87(m,2H),1.87–1.75(m,4H),1.72–1.51(m,6H),1.40–1.25(m,6H),1.13(d,J=6.7Hz,6H).

[0218] 18. Synthesis of Compound 18

[0219]

[0220] Following the synthetic route of compound 14, we obtained 18 52 mg of a white solid compound.

[0221] UPLC-MS calculated for C 48 H 53 ClN6O7[M+H] + :861.37,found:861.44.

[0222] 1H NMR (500MHz, DMSO-d6) δ10.08(s,1H),9.64(s,1H),8.46(t,J=6.8Hz,1H),8.01(d,J=11.5Hz,1H),7.89(s,4H),7.77(d,J= 7.4Hz,1H),7.59(d,J=0.6Hz,1H),7.30–7.22(m,2H),7.19–7.12(m,2H),6.99(dd,J=7.7,1.5Hz,1H),6.55(s,1H),4.53(d, J=1.0Hz,2H),4.48(d,J=1.1Hz,2H),4.18–4.09(m,1H),3.65–3.54(m,3H),3.49(t,J=7.1Hz,4H),3.33(q,J=7.0Hz,2H),3. 22–3.11(m,1H),2.47(t,J=7.1Hz,4H),2.36(t,J=7.1Hz,2H),1.95–1.73(m,8H),1.61–1.49(m,2H),1.13(d,J=6.7Hz,6H).

[0223] 19. Synthesis of Compound 19

[0224]

[0225] Following the synthetic route of compound 3, 19.39 mg of a white solid compound was obtained.

[0226] UPLC-MS calculated for C 46 H 52 F3N9O4[M+H] + :852.41,found:852.98.

[0227] 1H NMR (500MHz, DMSO-d6) δ10.08(s,1H),9.64(s,1H),7.92(d,J=7.5Hz,1H),7.78(d,J=7.4Hz,1H),7.59(d,J=0.6Hz,1H),7.51(d,J=7.5H z,1H),7.30–7.22(m,2H),7.19(d,J=1.5Hz,1H),7.15(dq,J=7.6,1.2Hz,1H),7.01(dd,J=7.5,1.5Hz,1H),6.55(s,1H),4.53(d,J=1.0H z,2H),4.48(d,J=1.1Hz,2H),3.73(t,J=7.2Hz,2H),3.70–3.61(m,6H),3.39–3.33(m,2H),3.31(t,J=7.1Hz,4H),3.22–3.11(m,1H),2. 65–2.60(m,2H),2.60–2.57(m,1H),2.57–2.54(m,5H),2.54–2.49(m,2H),2.49–2.44(m,1H),1.94–1.84(m,5H),1.12(d,J=6.7Hz,6H).

[0228] 20. Synthesis of Compound 20

[0229]

[0230] Following the synthetic route of compounds 2 and 3, 20-25 mg of a white solid compound was obtained.

[0231] UPLC-MS calculated for C 48 H 57 ClN 10 O4[M+H] + :873.42,found:873.50.

[0232] 1H NMR(500MHz,DMSO-d6)δ10.08(s,1H),9.64(s,1H),8.11(d,J=9.6Hz,1H),8.06(d,J =1.5Hz,1H),7.85(d,J=1.5Hz,1H),7.77(d,J=1.5Hz,1H),7.70(dd,J=17.0,7.5Hz,2 H),7.65–7.57(m,2H),7.30–7.22(m,2H),7.15(dq,J=7.6,1.2Hz,1H),6.66(d,J=7. 5Hz,1H),6.55(s,1H),4.53(d,J=1.0Hz,2H),4.48(d,J=1.1Hz,2H),4.35(dt,J=12.4 ,7.1Hz,1H),4.27(dt,J=12.4,7.1Hz,1H),4.15(dd,J=12.4,7.0Hz,1H),4.09–4.01 (m,1H),3.94(dh,J=9.8,6.9Hz,1H),3.63(t,J=1.0Hz,2H),3.22–3.11(m,1H),3.01( dt,J=12.4,7.2Hz,1H),2.95–2.84(m,5H),2.69(dd,J=12.4,7.0Hz,1H),2.60–2.48( m,9H),1.89–1.77(m,J=7.0Hz,1H),1.73–1.63(m,4H),1.11(dd,J=11.6,6.7Hz,9H).

[0233] Example 3: Bioevaluation Experiment

[0234] Example 1: Evaluation of the effect of reducing androgen receptor (AR) expression.

[0235] AR-positive human prostate cancer cells LNCaP were cultured in RPMI 1640 (hereinafter referred to as evaluation medium) containing 10% FBS until a concentration of 3 × 10⁻⁶ cells / mL was reached. 5The cells were seeded in 6-well microplates (Corning) and cultured overnight. Evaluation medium containing the compound from Example 2 was added to the culture to achieve a final concentration of 10 μmol / L, and the culture was continued for 24 hours. After 24 hours of culture, the medium was removed, the cells were washed with PBS, and RIPA lysis buffer containing 1% Protease Inhibitor Cocktail was added. After lysis and centrifugation, the total protein extract was obtained, and the protein concentration in the extract was detected by BCA method. Protein electrophoresis was performed by SDS-PAGE, and then the protein was transferred to a PVDF (MilliporeSigma IPVH00010) membrane by constant current electrophoresis at 200 mA for 90 min. The PVDF membrane was placed in 5% skim milk and blocked at room temperature for 1 h. Immunoreaction was performed using Anti-Androgen Receptor antibody [EPR1535(2)] (HRP) (abcam) and anti-AR-V7 Speccific antibody (Cell Signaling), respectively. After washing the membrane, ECL luminescent solution was added and the membrane was exposed. Grayscale analysis of the bands was performed using ImageJ software. The GAPDH protein band was simultaneously detected in each sample as an internal control. The degradation rate of the AR protein from the compound in the examples was calculated based on the grayscale values ​​of the protein bands.

[0236] The results for LNCaP and 22Rv1 cells are shown in Table 1. A reduction in AR expression of 50% or more is indicated as "reduction". In the compounds of this invention, compounds 1-20 all exhibited a certain degree of AR expression reduction.

[0237] Table 1: Effects of compounds on AR expression levels in LNCaP and 22Rv1 cells

[0238]

[0239]

[0240] Experimental Example 2: Inhibitory Activity on Androgen-Dependent Prostate Cancer Cell Proliferation

[0241] Androgen receptor-positive human prostate cancer cells LNCaP and 22RV1 were cultured in RPMI 1640 medium (hereinafter referred to as evaluation medium) containing 5% carbon-adsorbed serum (CCS) at a concentration of 4 × 10⁻⁶. 3The culture medium was seeded in 96-well microplates (Corning) with a clear bottom and cultured for 48 h. Evaluation medium containing R1881 (final concentration of R1881: 0.1 nmol / L) and evaluation medium containing compounds from Example 2 (final concentrations of 1.53, 4.6, 13.8, 41.1, 123.4, 370.3, 1111, 3333, and 10000 nmol / L) were added to the culture, and after 96 hours of culture, the number of viable cells was determined. The number of viable cells was determined using a WST-1 (Roche) assay. The cell proliferation activity value of 0.1 nmol / L R1881 was set as 100%, and the cell proliferation activity of the evaluation medium alone was set as 0%. The 50% inhibition concentration (IC50) was calculated using logistic regression based on the measured number of viable cells. 50 value).

[0242] The results of cell proliferation inhibition are shown in Table 2. This indicates that compounds 1-20 of the present invention exhibit certain prostate cancer cell proliferation inhibitory activity.

[0243] Table 2: Effects of compounds on the inhibitory activity of androgen-dependent prostate cancer cell proliferation.

[0244]

[0245]

[0246] The above embodiments are only used to illustrate the technical solutions of the present invention, and are not intended to limit them. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art can still modify the technical solutions described in the foregoing embodiments or make equivalent substitutions for some of the technical features. Such modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions claimed by the present invention.

Claims

1. A class of androgen receptor modulators characterized in that, The structural formula of the androgen receptor modulator is respectively: ； ； ； ； ； ； ； ； ； ； ； ； ； ； ； ； ； ； ； 。 2. A pharmaceutical composition, characterized by, The pharmaceutical composition comprises the compound or pharmaceutically acceptable salt thereof in claim 1.

3. The androgen receptor modulator in claim 1 or the pharmaceutical composition in claim 2 is used for preparing an anti-tumor drug.

4. Use according to claim 3, characterized in that, The tumor is prostate cancer.

5. Use according to claim 3, characterized in that, The drug can inhibit the proliferation of prostate cancer cells.

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