Conjugate of androgen receptor modulator and preparation method thereof

Abstract

The invention belongs to the technical field of medicines, and particularly relates to a conjugate serving as an androgen receptor modulator as well as a preparation method and application of the conjugate. The invention provides a conjugate molecule with a brand new structure, and the conjugate molecule is structurally characterized in that the conjugate molecule is an enzalutamide / apalutamide-connection structure Luminespib and a derivative, and can be used as an AR (Augmented Reality) regulator. Experiments show that the compound provided by the invention shows a certain AR protein degradation effect, and some compounds cause down-regulated AR protein expression gt; and 50%. Meanwhile, the compound also shows certain prostate cancer cell proliferation inhibition activity, and IC50 of prostate cancer cell proliferation inhibition activity of some compounds is less than 100nM.

Description

Technical Field

[0001] The present invention belongs to the field of medical technology, and in particular relates to a conjugate as an androgen receptor modulator, and a preparation method and application thereof. Background Art

[0002] Prostate cancer is one of the most common malignant tumors of the male genitourinary system, and its incidence ranks second among all male malignant tumors, and the incidence and mortality rates show a clear upward trend year by year. Since the early symptoms of prostate cancer are not obvious, most patients develop into advanced stages due to lack of regular screening. Currently, androgen deprivation therapy (ADT) is the first-line treatment for advanced prostate cancer. However, with the widespread use of this therapy, patients will inevitably develop into more difficult-to-cure castration-resistant prostate cancer (CRPC) within a few years.

[0003] Androgen receptor (AR) and its downstream signal transduction play an important role in the occurrence and development of CRPC. Inhibiting androgen production or blocking the binding of androgen to AR through drugs is an effective means of treating CRPC. Currently, small molecule antagonists targeting AR on the market include apalutamide, enzalutamide, darolutamide, etc. However, the gradual emergence of drug resistance in patients during treatment has put more and more CRPC patients at risk of having no drugs available.

[0004] Protein degradation technology is an emerging drug development strategy in recent years. It achieves the purpose of treating diseases by degrading pathogenic proteins. It has the characteristics of low-volume and high-efficiency, and is expected to solve the problem of traditional small molecule inhibitors prone to drug resistance. It treats diseases by directly regulating the protein content in the body, providing a broad new idea for innovative drug research. However, as of now, there are no protein degradation drugs on the market. Therefore, this field still needs to develop androgen receptor modulators that can induce AR degradation, and / or compounds with better pharmacodynamics / pharmacokinetics. Summary of the invention

[0005] In view of the existing problems, the present invention provides a class of conjugate molecules with a new structure, which can be used as AR regulators to significantly inhibit the proliferation of prostate cancer cells by reducing the expression level of AR.

[0006] The present invention provides a compound represented by formula I, its optical isomers, isotopic derivatives or pharmaceutically acceptable salts thereof,

[0007]

[0008] In Formula I, Ring A is selected from C5-C10 heteroaryl; Ring B is selected from C4-10 C ring is selected from C6-C10 membered aryl, C5-C10 membered heteroaryl;

[0009] X, Y, Z are C or N;

[0010] R 1 , R 2 , R 3 They are independently selected from H, halogen, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl, OH, NH2, CN, nitro, carboxyl, and C3-6 cycloalkyl; and the C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl, OH, NH2, carboxyl, and C3-6 cycloalkyl may contain 1, 2 or 3 substituents selected from halogen, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl, OH, NH2, CN, nitro, and carboxyl, respectively.

[0011] R 4 Selected from H, halogen, C1-C6 alkyl, C1-C6 haloalkyl;

[0012] R 5 , R 6 are each independently selected from H, halogen, C1-C6 alkyl, C1-C6 haloalkyl, and R 3 and R 4 Connected to form a ring with the carbon atom shared by both;

[0013] R 7 , R 8 Selected from H, halogen, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl, OH, NH2, CN, nitro, carboxyl, C3-C6 cycloalkyl;

[0014] L 1 , L 2 , L 3 , L 4 and L 5 Each independently and compatibly selected from a single bond, a C1-C6 alkylene group, a carbonyl group, a 4-10 membered heterocycloalkylene group, -O-, -N(R B1 )R B2 -, where R B1 is H, C1-C6 alkyl, R B2 is a C0-C6 alkylene group;

[0015] Preferably, in Formula I, Ring A is selected from

[0016] Ring B is selected from The C ring is selected from X is C or N; Y is C; Z is C; R 1 Selected from -OH; R 2 Selected from -OH; R 3 Selected from R 4 Selected from -CF3; R 5 Selected from -CH3 or -CH2CH2-, one side of which is connected to R 6 Connect; R 6 Selected from -CH3 or -CH2CH2-, one side of which is connected to R 5 Connect; R 7 Selected from -F;R 8 Selected from -H; Selected from

[0017] More preferably, the compound has any one of the following structural formulas: Compound 1: Compound 2: Compound 3: Compound 4: Compound 5: Compound 6: Compound 7: Compound 8: Compound 9: Compound 10: Compound 11: Compound 12: Compound 13: Compound 14: Compound 15: Compound 16:

[0018] The present invention also provides a method for preparing the compound.

[0019] The present invention provides a new type of conjugate molecule, whose structural features are enzalutamide / apalutamide-connector structure-Luminespib and derivatives, which achieve the regulation of AR function and content through the synergistic effect of dual targets. Experiments show that the compounds provided by the present invention exhibit a certain AR protein degradation effect, and some compounds cause AR protein expression to be downregulated by >50%. At the same time, they also exhibit a certain prostate cancer cell proliferation inhibition activity, and some compounds have an IC of prostate cancer cell proliferation inhibition activity of >100%. 50 Less than 100nM. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 The proliferation inhibitory activity of some of the compounds in the examples on VCaP cells.

[0021] Figure 2 The proliferation inhibitory activity of some of the compounds in the examples on 22Rv1 cells. DETAILED DESCRIPTION

[0022] The present invention is further described below through specific examples in order to better understand the present invention, but does not constitute a limitation of the present invention.

[0023] Example 1: Synthesis of intermediate S10

[0024] The synthetic route of intermediate S10 is as follows:

[0025]

[0026] The specific synthesis steps are as follows:

[0027] Step 1: Add the raw material S1 (5 g, 32.8 mmol) to a 100 mL reaction bottle, add boron trifluoride etherate (28 g, 197.1 mmol) and acetic acid (3.9 g, 65.7 mmol), replace with argon three times, stir at 90 ° C overnight, add 10% NaOAc to neutralize to pH = 7 under ice bath, stir for 10 min, extract with ethyl acetate (200 mL * 2), wash the organic phase with saturated brine (100 mL * 2), dry over anhydrous sodium sulfate, filter, and concentrate to obtain 6 g of crude product, which was purified by column chromatography (PE: EA = 20: 1 ~ 10: 1) to obtain 5.4 g of light yellow solid, with a yield of 84.6%. UPLC-MS calculated for C 11 H 14 O3[M+H] + :195.09,found:195.11.

[0028] Step 2: Add S2 (5.4 g, 27.8 mmol), K2CO3 (11.5 g, 83.5 mmol) and DMF (200 mL) into a 500 mL reaction bottle, add benzyl bromide (10.5 g, 61.2 mmol) dropwise, replace with argon three times, heat to 110 ° C and stir overnight. Pour the reaction solution into 1.5 L ice water, stir for 30 min, filter the precipitated solid and wash with 300 mL water, dry to obtain 11 g crude product. Recrystallize with MeOH / DCM to obtain 9.1 g white solid, yield 87.4%. UPLC-MS calculated for C 25 H 26 O3[M+H] + :375.19,found:375.23.

[0029] Step 3: Add S3 (6.5 g, 17.4 mmol) to an ethanol (100 mL) solution of EtONa (2.4 g, 34.7 mmol), replace with argon three times, stir for 10 min, add diethyl oxalate (3.9 g, 26.1 mmol) dropwise, heat and reflux and stir for 4 h. After cooling to room temperature, neutralize with 1 M HCl to pH = 7, filter the precipitated solid, wash the filter cake with cold methanol (20 mL), and dry to obtain 7 g of yellow solid, with a yield of 84.9%. UPLC-MS calculated for C 29 H 30 O6[M+H] + :475.20,found:475.23.

[0030] Step 4: S4 (7 g, 14.8 mmol), hydroxylamine hydrochloride (1.2 g, 17.7 mmol) and ethanol (80 mL) were added to the reaction bottle, replaced with argon three times, heated to reflux and stirred for 3 h, cooled in an ice bath, stirred for 30 min, filtered, the filter cake was washed with cold ethanol (20 mL), and dried to obtain 5 g of white solid, with a yield of 71.4%. UPLC-MS calculated for C29H29NO5[M+H] + :472.20,found:472.23.

[0031] Step 5: Add S5 (5.3 g, 11.2 mmol), NBS (2 g, 11.2 mmol) and acetonitrile (70 mL) to the reaction bottle, add cerium ammonium nitrate (0.06 g, 0.1 mmol), replace the argon three times, heat and reflux and stir overnight. The reaction solution was cooled to room temperature and concentrated to obtain a crude product. Column chromatography (PE: EA = 19: 1) gave 5.6 g of a white solid, with a yield of 90.4%. UPLC-MS calculated for C 29 H28 BrNO5[M+H] + :550.12, 552.11, found:550.14, 552.15.

[0032] Step 6: Dissolve S6 (5.1 g, 9.3 mmol), 4-acylphenylboronic acid (1.67 g, 11.1 mmol), and Na2CO3 (3.4 g, 27.8 mmol) in 20 mL of water and add THF (70 mL) to the reaction bottle, replace the argon three times, add Pd(TPP)2Cl2 (0.33 g, 0.46 mmol), replace the argon three times again, heat and reflux and stir overnight. The reaction solution was cooled to room temperature, 100 mL of water was added, and extracted with ethyl acetate (100 mL*2). The organic phase was washed with saturated brine (100 mL), dried over anhydrous Na2SO4, filtered and concentrated to obtain a crude product. Purification by column chromatography (PE:EA=85%:15%) gave 3.8 g of a yellow solid with a yield of 64.9%. UPLC-MS calculated for C 36 H 33 NO6[M+H] + :576.23,found:576.25.

[0033] Step 7: Add S7 (2g, 3.5mmol), morpholine (610mg, 6.9mmol), acetic acid (310mg, 5.2mmol) and methanol (30mL) to the reaction bottle, stir at room temperature for 3h, add NaBH3CN (330mg, 5.2mmol), and stir for 1h. Add 100mL of water to quench the reaction, extract with ethyl acetate (100mL*2), wash the organic phase with saturated brine (100mL), dry with anhydrous Na2SO4, filter, and concentrate to obtain a crude product. Purify by column chromatography (DCM:MeOH=49:1) to obtain 1.5g of white solid with a yield of 60.7%. UPLC-MS calculated for C 40 H 42 N2O6[M+H] + :647.30,found:647.31.

[0034] Step 8: Add S8 (1.5 g, 2.3 mmol) and dichloromethane (20 mL) to the reaction bottle, replace the argon three times, add BCl3 (13.9 mL, 13.9 mmol, 1 M toluene solution) dropwise in an ice-water bath, and stir in an ice-water bath for 2 h. Add water (50 mL) in an ice-water bath to quench, extract with dichloromethane (50 mL*2), wash the organic phase with saturated brine (50 mL), dry over anhydrous Na2SO4, filter and concentrate to obtain 1 g of light brown solid, with a yield of 92.7%. UPLC-MS calculated for C 26 H 30 N2O6[M+H] + :466.21,found:466.24.

[0035] Step 9: S9 (1 g, 2.14 mmol) was dissolved in THF (10 mL), and a solution of LiOH (0.3 g, 6.44 mmol) in water (2 mL) was added dropwise at room temperature and stirred for 3 h. The pH was adjusted to 3 with 1 M HCl, and the aqueous phase was directly freeze-dried to obtain 1.3 g of crude product, which was dissolved in acetonitrile (20 mL), filtered and concentrated to obtain 800 mg of light brown solid S10, with a yield of 85.1%. UPLC-MS calculated for C 24 H 26 N2O6[MH] - :435.18,found:435.22.

[0036] Example 2: Synthesis of Intermediate S16

[0037] The synthetic route of intermediate S16 is as follows:

[0038]

[0039] The specific synthesis steps are as follows:

[0040] Step 1: S4 (2.7 g, 5.7 mmol), hydrazine hydrate (0.43 g, 6.8 mmol, 80%) and acetic acid (30 mL) were added to the reaction bottle, argon was replaced three times, stirred at 85 ° C overnight, cooled to room temperature, poured into 500 mL of water, stirred for 30 min, filtered, and the filter cake was washed with cold ethanol (20 mL) and dried to obtain 2.6 g of white solid S11, with a yield of 97%. UPLC-MS calculated for C 29 H 30 N2O4[M+H] + :471.22,found:471.25.

[0041] Step 2: Add S11 (2.6 g, 5.5 mmol), NBS (1 g, 5.6 mmol) and acetonitrile (40 mL) into a reaction flask, add cerium ammonium nitrate (0.3 g, 0.55 mmol), replace with argon three times, heat and reflux and stir overnight. The reaction solution was cooled to room temperature and concentrated to obtain a crude product. Column chromatography (PE:EA=19:1) gave 2.6 g of light yellow solid S12, yield: 85%. UPLC-MS calculated for C 29 H 29 BrN2O4[M+H] + :549.13, 551.13,found:549.14, 551.15.

[0042] Step 3: Add S12 (600 mg, 1.09 mmol), 4-formylphenylboronic acid (240 mg, 1.6 mmol), K3PO4 (300 mg, 3.3 mmol) and 1,4-dioxane (10 mL) to the reaction bottle, replace the argon three times, add Pd(dtbpf)Cl2 (84 mg, 0.11 mmol), replace the argon three times again, and stir at 95°C overnight. The reaction solution was cooled to room temperature, 100 mL of water was added, extracted with ethyl acetate (100 mL*2), the organic phase was washed with saturated brine (100 mL), dried over anhydrous Na2SO4, filtered and concentrated to obtain a crude product. Purified by column chromatography (DCM:EA=4:1) to obtain 300 mg of brown solid with a yield of 47%. UPLC-MS calculated for C 36 H 34 N2O5[M+H] + :575.25,found:575.68.

[0043] Step 4: Add S13 (300 mg, 0.52 mmol), morpholine (91 mg, 1.04 mmol), acetic acid (47 mg, 0.78 mmol) and methanol (20 mL) to the reaction bottle, stir at room temperature for 3 h, add NaBH3CN (49 mg, 0.78 mmol), and stir for 1 h. Add 100 mL of water to quench the reaction, extract with ethyl acetate (50 mL*2), wash the organic phase with saturated brine (50 mL), dry over anhydrous Na2SO4, filter, and concentrate to obtain a crude product. Purify by column chromatography (DCM: MeOH = 49: 1) to obtain 220 mg of a white solid with a yield of 65%. UPLC-MS calculated for C 40 H 43 N3O5[M+H] + :646.32,found:646.80.

[0044] Step 5: S14 (200 mg) and acetic acid (5 mL) were added to the reaction flask, and 10% Pd / C (20 mg) was added. The mixture was stirred overnight under hydrogen, filtered and concentrated, and purified by column chromatography (DCM:MeOH=20:1) to obtain 65 mg of a light yellow solid with a yield of 45%. UPLC-MS calculated for C 26 H 31 N3O5[M+H] + :466.23,found:466.55.

[0045] Step 6: S15 (60 mg, 0.13 mmol) and 1,4-dioxane were added to a reaction flask, and a 20% aqueous solution of NaOH (10 mg, 0.25 mmol) was added, stirred at room temperature overnight, 20 mL of water was added, and the solution was neutralized to pH = 5 with 1M HCl, extracted with ethyl acetate (50 mL*2), and the organic phase was washed with saturated brine (50 mL), dried over anhydrous Na2SO4, filtered and concentrated to obtain 40 mg of light yellow solid S16. UPLC-MS calculated for C 24 H 27 N3O5[MH] - :436.20,found:436.50.

[0046] Example 3: Synthesis of intermediate S19

[0047] The synthetic route of intermediate S19 is as follows:

[0048]

[0049] The synthesis method refers to the synthesis steps of intermediate S10 in Example 1, wherein the raw material is replaced by methylpiperazine instead of morpholine to obtain 60 mg of a light yellow solid. UPLC-MS calculated for C 25 H 29 N3O5[MH] - :450.21,found:450.52.

[0050] Example 4: Synthesis of Intermediate S25

[0051] The synthetic route of intermediate S25 is as follows:

[0052]

[0053] The specific synthesis steps are as follows:

[0054] Step 1: Dissolve the raw material S20 (2.0 g, 9.26 mmol) in dichloromethane (60 mL), add morpholine (819 mg, 9.4 mmol) and triethylamine (2 mL, 14.0 mmol). The reaction mixture was stirred at room temperature for 6 h, extracted with water (100 mL*3), and the organic phase was washed with saturated brine and dried over anhydrous sodium sulfate. After removing the solvent, recrystallization was performed with ethanol to obtain 1.66 g of a light yellow solid with a yield of 81%. UPLC-MS calculated for C 11 H 14 N2O3[M+H] + :223.10,found:223.24.

[0055] Step 2: Intermediate S21 (1.5 g, 6.75 mmol) and stannous chloride dihydrate (7.62 g, 33.77 mmol) were placed in ethyl acetate (50 mL) and stirred at room temperature overnight. Saturated sodium bicarbonate solution (20 mL) was added and stirred vigorously for 1 h. The solid was removed by filtration, extracted with ethyl acetate (100 mL*2), and the organic phase was washed with saturated brine (100 mL). The organic phase was dried over anhydrous magnesium sulfate and concentrated to obtain 1.02 g of solid, with a yield of 78%. UPLC-MS calculated for C 11 H 16 N2O[M+H] + :193.13,found:193.26.

[0056] Step 3: Dissolve the intermediate 2,4-dihydroxy-5-isopropylbenzodithioic acid (430.92 mg, 1.89 mmol) in DMF (5 mL), add NaHCO3 (577.31 mg, 6.87 mmol), add chloroacetic acid (162.37 mg, 1.17 mmol) under argon protection at 0°C, return to room temperature and stir for 2 h, dissolve the intermediate S22 (500 mg, 1.71 mmol) in DMF and add to the system, react at 80°C overnight, cool to room temperature, add ammonium chloride aqueous solution, extract with ethyl acetate, evaporate the organic phase solvent, separate by column chromatography (PE:EA=1:1), and obtain 350 mg of yellow solid, with a yield of 48%. UPLC-MS calculated for C 21 H 26 N2O3S[M+H] + :387.17,found:387.51.

[0057] Step 4: Dissolve the intermediate S23 (350 mg, 0.91 mmol) in 1,4-dioxane (15 mL), add hydrazine hydrate (206.18 mg, 4.13 mmol), stir and react at 80 °C for 5 h under argon protection, cool to room temperature, evaporate the solvent to obtain 320 mg of yellow oil. UPLC-MS calculated for C 21 H 28 N4O3[M+H] + :385.22,found:385.48.

[0058] Step 5: Dissolve the intermediate S24 (300 mg, 0.78 mmol) in EtOH (10 mL), add two drops of AcOH, dissolve diethyl oxalate (171 mg, 1.17 mmol) in ethanol and add dropwise to the above system at 0°C, return to room temperature under argon protection and stir overnight, evaporate the solvent, separate by column chromatography (PE:EA=2:1), and obtain 131 mg of white solid, with a yield of 36%. UPLC-MS calculated for C 25 H 30 N4O5[M+H] + :467.22,found:467.54.

[0059] Example 5: Synthesis of Intermediate S30

[0060] The synthetic route of intermediate S30 is as follows:

[0061]

[0062] The specific synthesis steps are as follows:

[0063] Step 1: Add raw material S26 (5 g, 21.5 mmol), 1-aminocyclobutanecarboxylic acid (3 g, 26 mmol) and 50 mL DMSO to a reaction bottle, replace the atmosphere with argon three times, add CuI (0.82 g, 4.3 mmol) and DBU (7.6 g, 50 mmol), replace the atmosphere with argon three times again, heat to 90 ° C and stir for 5 h, cool and pour the reaction solution into 200 mL of water, extract with ethyl acetate (50 mL*2) and discard, acidify the aqueous phase with 2M HCl to pH = 2, filter the solid, wash the filter cake with 200 mL of water, and dry to obtain 4 g of white solid. UPLC-MS calculated for C 13 H 14 FNO4[MH] - :266.09,found:266.26.

[0064] Step 2: Add the raw material S27 (3.4 g, 12.7 mmol) and 50 mL MeOH into the reaction bottle, add concentrated sulfuric acid (1.37 g, 14.0 mmol), replace the argon three times, heat to reflux and stir for 4 h, and after cooling, neutralize the reaction solution with NaHCO3 to pH = 7, filter the solid, wash the filter cake with 50 mL water and 50 mL cold MeOH, and dry to obtain 3.2 g of light brown solid. UPLC-MS calculated for C 14 H 16 FNO4[M+H] + :282.11,found:282.28.

[0065] Step 3: Add raw material S28 (2.5 g, 8.9 mmol) and 15 mL THF into the reaction bottle, replace the argon three times, add LiHMDS (1 Min THF, 10 mL, 9.8 mmol) dropwise under ice-water bath, stir for 30 min, add 10 mL THF solution of 5-isothiocyanato-3-(trifluoromethyl)pyridine-2-cyano (2.2 g, 9.3 mmol) dropwise into the reaction system, stir for 1 h under ice-water bath, acidify the reaction solution with 2 M HCl to pH = 5, extract with ethyl acetate (50 mL * 2), wash the organic phase with 50 mL saturated brine, dry, filter and concentrate over anhydrous sodium sulfate to obtain 4.5 g light brown solid S29, UPLC-MS calculated for C 21 H 14 F4N4O3S[M+H] + :479.07,found:479.42.

[0066] Step 4: S29 (4.5 g, 9.4 mmol) and THF were added to a reaction flask, and a 20% aqueous solution of LiOH (675 mg, 28.20 mmol) was added, stirred at room temperature overnight, 20 mL of water was added, pH was adjusted to 5 with 1 M HCl, and extracted with ethyl acetate (50 mL*2), and the organic phase was washed with saturated brine (50 mL), dried over anhydrous Na2SO4, filtered and concentrated to obtain 3.71 g of solid crude S30. UPLC-MS calculated for C 20 H 12 F4N4O3S[MH] - :463.06,found:463.39.

[0067] Example 6: Synthesis of Compound 1

[0068] The synthetic route of compound 1 is as follows:

[0069]

[0070] The specific synthesis steps are as follows:

[0071] Step 1: Add raw material S10 (200 mg, 0.46 mmol), N-Boc-piperazine (128 mg, 0.68 mmol), HOBt (100 mg, 0.68 mmol), DIEA (200 mg, 1.4 mmol) and DMF (10 mL) to the reaction bottle, add PyBOP (353 mg, 0.68 mmol), replace with argon three times, stir at room temperature overnight, add 50 mL of water, extract with ethyl acetate (50 mL*2), wash the organic phase with saturated brine (50 mL), dry with anhydrous sodium sulfate, filter and concentrate to obtain a crude product. Purify by column chromatography (DCM: MeOH = 20: 1) to obtain 180 mg of white solid, with a yield of 60%. UPLC-MS calculated for C 33 H 42 N4O7[M+H] + :607.31,found:607.51.

[0072] Step 2: Add S31 (180 mg, 0.297 mmol) to the reaction flask, add 4M HCl / 1,4-dioxane solution (0.74 mL, 2.97 mmol) dropwise at room temperature, and stir for 1 h. Add 2 mL of methanol, filter, concentrate, and pump dry to obtain 160 mg of white solid, with a yield of 100%. UPLC-MS calculated for C 28 H 34 N4O5[M+H] + :507.25,found:507.60.

[0073] Step 3: Dissolve the raw material S33 (4.64 g, 10 mmol) in 1,4-dioxane (50 mL), add concentrated HCl (5 mL), stir the reaction at 110 ° C for 4 h, and monitor the reaction by thin layer chromatography (TLC). The reaction mixture was cooled to ambient temperature, quenched by adding water (50 mL), extracted with dichloromethane (100 mL*2), and the organic phase was washed with saturated brine (100 mL). The organic phase was dried over anhydrous magnesium sulfate, and the solvent was evaporated and separated by column chromatography (Hexane: EA = 4: 1) to obtain 4.06 g of solid, with a yield of 90%. UPLC-MS calculated for C 20 H 13 F4N3O3S[M+H] + :452.04,found:452.40.

[0074] Step 4: S34 (150 mg, 0.33 mmol), DIEA (86 mg, 0.66 mmol) and DMF (10 mL) were added to the reaction flask, argon was replaced three times, HATU (189 mg, 0.50 mmol) was added and stirred at room temperature for 1 h, intermediate S32 (190 mg, 0.35 mmol) was added and stirred at room temperature overnight, and 102 mg of white solid was obtained by preparative liquid separation, with a yield of 33%. UPLC-MS calculated for C 48 H 45 F4N7O7S[M+H] + :940.30,found:940.98. 1 H NMR (400MHz, DMSO-d6) δ9.81(s,1H),9.70(s,1H),8.90(t,J=5.8Hz,1H),8.54(t,J=5.7Hz,1H),8.41(d,J=8 .3Hz,1H),8.30(d,J=1.9Hz,1H),8.08(dd,J=8.3,1.9Hz,1H),7.79(t,J=8.0Hz,1H),7.44(dd,J=10.6,1.9H z,1H),7.34(dd,J=8.1,1.9Hz,1H),7.21(q,J=8.0Hz,4H),6.72(s,1H),6.44(s,1H),3.73(s,8H),3.54(t,J =4.5Hz, 4H), 3.40 (s, 2H), 2.96 (p, J = 6.9Hz, 1H), 2.31 (t, J = 4.4Hz, 4H), 1.54 (s, 6H), 0.89 (d, J = 6.9Hz, 6H).

[0075] Example 7: Synthesis of Compound 2

[0076] The synthetic route of compound 2 is as follows:

[0077]

[0078] The specific synthesis steps are as follows:

[0079] Step 1: Add S34 (150 mg, 0.33 mmol), DIEA (86 mg, 0.66 mmol) and DCM (10 mL) to a reaction flask, replace the argon atmosphere three times, add HATU (189 mg, 0.50 mmol) and stir at room temperature for 1 h, add 4-piperidinemethanol (41 mg, 0.36 mmol) and stir at room temperature overnight, add water (50 mL), extract with ethyl acetate (50 mL*2), wash the organic phase with saturated brine (50 mL), dry over anhydrous sodium sulfate, filter and concentrate to obtain a crude product, and obtain 132 mg of solid by column chromatography (DCM: MeOH = 20: 1), with a yield of 73%. UPLC-MS calculated for C 26 H 24 F4N4O3S[M+H] + :549.15,found:549.56.

[0080] Step 2: Dissolve the intermediate S35 (100 mg, 0.18 mmol) in dichloromethane (20 mL), slowly add DMP (155 mg, 0.36 mmol) into the reaction bottle, and stir at room temperature for 2 h. TLC detected that the raw material reaction was complete, and the organic phase was washed with saturated sodium bicarbonate and saturated brine in turn, dried over anhydrous sodium sulfate, and the solvent was evaporated and column chromatography (DCM: MeOH = 50: 1) was performed to obtain 84 mg of a light yellow solid, with a yield of 86%. UPLC-MS calculated for C 26 H 22 F4N4O3S[M+H] + :547.13,found:547.54.

[0081] Step 3: S36 (80 mg, 0.15 mmol) and MeOH (3 mL) were added to a reaction flask, triethylamine (23 mg, 0.22 mmol) was added, and the mixture was stirred at room temperature for 30 min. S32 (61 mg, 0.15 mmol) and acetic acid (27 mg, 0.45 mmol) were added in sequence, and the mixture was stirred at room temperature for 5 h. NaBH3CN (18 mg, 0.30 mmol) was added, and the mixture was stirred for 1 h. After filtration, 31 mg of a white solid was obtained by reverse phase preparative purification, with a yield of 20%. UPLC-MS calculated for C 54 H 56 F4N8O7S[M+H] + :1037.39,found:1037.42. 1H NMR (400MHz, DMSO-d6) δ9.71(s,1H),9.70(s,1H),8.90(t,J=5.8Hz,1H),8.54(t,J=5.7Hz,1H),8.41(d,J=8.3Hz,1H),8.30(d,J=1. 9Hz,1H),8.08(dd,J=8.3,1.9Hz,1H),7.79(t,J=8.0Hz,1H),7.44(dd,J=10.6,1.9Hz,1H),7.34(dd,J=8.1,1.9Hz,1H),7.21(q,J=8. 0Hz,4H),6.72(s,1H),6.44(s,1H),4.44(d,J=12.8Hz,2H),3.59(s,2H),3.55(t,J=4.6Hz,4H),3.40(s,2H),3.35(s,8H),3.12–2.98 (m,4H),2.96(p,J=6.9Hz,1H),2.31(t,J=4.4Hz,4H),2.10–2.00(m,1H),1.78(d,J=11.5Hz,2H),1.54(s,6H),0.96(d,J=6.9Hz,6H).

[0082] Example 8: Synthesis of Compound 3

[0083] The synthetic route of compound 3 is as follows:

[0084]

[0085] The specific synthesis steps are as follows:

[0086] The synthesis method was similar to that of Example 7. The raw material was replaced by 4-hydroxypiperidine from 4-piperidinol to obtain 11 mg of a white solid. UPLC-MS calculated for C 53 H 54 F4N8O7S[M+H] + :1023.38,found:1024.12. 1H NMR (400MHz, DMSO-d6) δ9.86(d,J=94.0Hz,2H),8.90(t,J=5.8Hz,1H),8.54(t,J=5.7Hz,1H),8.41(d,J=8.3Hz,1H),8.30(d,J =1.9Hz,1H),8.08(dd,J=8.3,1.9Hz,1H),7.79(t,J=8.0Hz,1H),7.44(dd,J=10.6,1.9Hz,1H),7.34(dd,J=8.1,1.9Hz,1H),7. 21(q,J=8.0Hz,4H),6.72(s,1H),6.44(s,1H),4.41(d,J=13.0Hz,2H),3.59(s,2H),3.52(t,J=4.5Hz,8H),3.40(s,2H),3.35( s,4H),3.14(d,J=4.9Hz,2H),3.02–2.82(m,3H),2.30(t,J=4.4Hz,4H),2.10–2.00(m,1H),1.54(s,6H),0.90(d,J=6.9Hz,6H).

[0087] Example 9: Synthesis of Compound 4

[0088] The synthetic route of compound 4 is as follows:

[0089]

[0090] The specific synthesis steps are as follows:

[0091] The synthesis method was as described in Example 7. The raw material was replaced by azetidine-3-ylmethanol from 4-piperidinol to obtain 25 mg of a white solid. UPLC-MS calculated for C 52 H 52 F4N8O7S[M+H] + :1009.36,found:1009.48. 1H NMR (400MHz, DMSO-d6) δ9.79(s,1H),9.65(s,1H),8.90(t,J=5.8Hz,1H),8.54(t,J=5.7Hz,1H),8.41(d,J=8.3Hz,1H),8. 30(d,J=1.9Hz,1H),8.08(dd,J=8.3,1.9Hz,1H),7.79(t,J=8.0Hz,1H),7.44(dd,J=10.6,1.9Hz,1H),7.34(dd,J=8.1,1. 9Hz, 1H), 7.21 (q, J = 8.0Hz, 4H), 6.72 (s, 1H), 6.44 (s, 1H), 4.41 (d, J = 13.0Hz, 2H), 3.59 (s, 2H), 3.52 (t, J = 4.5Hz, 8H), 3. 40(s,2H),3.35(s,4H),2.98–2.94(m,1H),2.30(t,J=4.4Hz,4H),2.10–2.00(m,1H),1.54(s,6H),0.90(d,J=6.9Hz,6H).

[0092] Example 10: Synthesis of Compound 5

[0093] The synthetic route of compound 5 is as follows:

[0094]

[0095] The specific synthesis steps are as follows:

[0096] The synthesis method refers to Examples 6 and 7, the raw material is replaced by 1-Boc-4-aminopiperidine from N-Boc-piperazine, and 21 mg of white solid is obtained. UPLC-MS calculated for C 55 H 58 F4N8O7S[M+H] + :1051.41,found:1051.62. 1H NMR (400MHz, DMSO-d6) δ9.75(s,2H),8.90(t,J=5.8Hz,1H),8.54(t,J=5.7Hz,1H),8.41(d,J=8.3Hz,1H),8.32(s,1H),8.30(d,J=1 .9Hz,1H),8.08(dd,J=8.3,1.9Hz,1H),7.79(t,J=8.0Hz,1H),7.44(dd,J=10.6,1.9Hz,1H),7.34(dd,J=8.1,1.9Hz,1H),7.21(q,J =8.0Hz,4H),6.72(s,1H),6.44(s,1H),4.41(d,J=13.0Hz,2H),3.59(s,2H),3.52(t,J=4.5Hz,8H),3.40(s,2H),3.35(s,5H),2.98 –2.94(m,1H),2.30(t,J=4.4Hz,4H),2.10–2.00(m,1H),1.95–1.84(m,2H),1.70–1.58(m,2H),1.54(s,6H),0.90(d,J=6.9Hz,6H).

[0097] Example 11: Synthesis of Compound 6

[0098] The synthetic route of compound 6 is as follows:

[0099]

[0100] The specific synthesis steps are as follows:

[0101] Step 1: S34 (150 mg, 0.33 mmol), DIEA (86 mg, 0.66 mmol) and DCM (10 mL) were added to the reaction flask, argon was replaced three times, HATU (189 mg, 0.50 mmol) was added and stirred at room temperature for 1 h, N-Boc-piperazine (67 mg, 0.36 mmol) was added and stirred at room temperature overnight, water (50 mL) was added, and ethyl acetate (50 mL*2) was used for extraction, the organic phase was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated to obtain a crude product, and 137 mg of solid was obtained by column chromatography (DCM: MeOH = 100: 1), with a yield of 67%. UPLC-MS calculated for C 29 H 29 F4N5O4S[M+H] + :620.19,found:620.64.

[0102] Step 2: S43 (150 mg, 0.33 mmol) and DCM (10 mL) were added to a reaction flask, and 4 M HCl / 1,4-dioxane solution (0.25 mL, 0.99 mmol) was added and stirred at room temperature for 2 h. The crude product S44 (183 mg) was obtained by concentration, with a yield of 99%. UPLC-MS calculated for C 24 H 21 F4N5O2S[M+H] + :520.14,found:520.52.

[0103] Step 3: S44 (150 mg, 0.27 mmol) and MeOH (3 mL) were added to a reaction flask, triethylamine (0.06 mL, 0.41 mmol) was added, and the mixture was stirred at room temperature for 30 min. N-Boc-piperidine-4-carboxaldehyde (86 mg, 0.41 mmol) and acetic acid (0.05 mL, 0.82 mmol) were added in sequence, and the mixture was stirred at room temperature for 5 h. NaBH3CN (34 mg, 0.54 mmol) was added, and the mixture was stirred for 1 h. After concentration, 137 mg of solid was obtained by column chromatography (DCM: MeOH = 50: 1), and the yield was 56%. UPLC-MS calculated for C 35 H 40 F4N6O4S[M+H] + :717.28,found:717.80.

[0104] Step 4: S45 (130 mg, 0.18 mmol) and DCM (10 mL) were added to a reaction flask, and 4 M HCl / 1,4-dioxane solution (0.18 mL, 0.73 mmol) was added and stirred at room temperature for 2 h. The crude product S46 (111 mg) was obtained by concentration, with a yield of 95%. UPLC-MS calculated for C 30 H 32 F4N6O2S[M+H] + :617.22,found:617.68.

[0105] Step 5: Add intermediate S10 (65 mg, 0.15 mmol), intermediate S46 (100 mg, 0.15 mmol), HOBt (31 mg, 0.23 mmol), DIEA (0.08 mL, 0.45 mmol) and DMF (50 mL) to a reaction bottle, add PyBOP (120 mg, 0.23 mmol), replace the argon gas three times, stir at room temperature overnight, filter and concentrate, and purify by reverse phase preparation to obtain 39 mg of white solid, with a yield of 25%. UPLC-MS calculated for C 54 H 56F4N8O7S[M+H] + :1037.39,found:1037.66. 1 H NMR (400MHz, DMSO-d6) δ9.68(s,2H),8.90(t,J=5.8Hz,1H),8.52(t,J=5.7Hz,1H),8.39(d,J=8.3Hz,1H),8.30(d,J=1.9Hz,1H),8 .07(dd,J=8.3,1.9Hz,1H),7.77(t,J=8.0Hz,1H),7.42(dd,J=10.6,1.9Hz,1H),7.31(dd,J=8.1,1.9Hz,1H),7.20(q,J=8.0Hz,4H ),6.72(s,1H),6.43(s,1H),4.42(d,J=12.8Hz,2H),3.59(s,2H),3.55(t,J=4.6Hz,4H),3.40(s,2H),3.35(s,8H),3.12–2.98(m, 4H), 2.96 (p, J = 6.9Hz, 1H), 2.31 (t, J = 4.4Hz, 4H), 2.10–2.00 (m, 1H), 1.78 (d, J = 11.5Hz, 2H), 1.54 (s, 6H), 0.96 (d, J = 6.9Hz, 6H).

[0106] Example 12: Synthesis of Compound 7

[0107] The synthetic route of compound 7 is as follows:

[0108]

[0109] The specific synthesis steps are as follows:

[0110] The synthesis method was based on Example 7, and the reverse preparation and purification gave 29 mg of a white solid. UPLC-MS calculated for C 49 H 47 F4N7O7S[M+H] + :954.32,found:954.46. 1H NMR (400MHz, DMSO-d6) δ9.75(s,2H),8.90(t,J=5.8Hz,1H),8.54(t,J=5.7Hz,1H),8.41(d,J=8.3Hz,1H),8.32(s,1H),8.30 (d,J=1.9Hz,1H),8.08(dd,J=8.3,1.9Hz,1H),7.79(t,J=8.0Hz,1H),7.44(dd,J=10.6,1.9Hz,1H),7.34(dd,J=8.1,1.9Hz,1 H),7.21(q,J=8.0Hz,4H),6.72(s,1H),6.44(s,1H),4.41(d,J=13.0Hz,2H),3.52(t,J=4.5Hz,4H),3.35(s,4H),3.02–2.94 (m,2H),2.30(t,J=4.4Hz,4H),2.10–2.00(m,1H),1.95–1.84(m,2H),1.70–1.58(m,2H),1.54(s,6H),0.90(d,J=6.9Hz,6H).

[0111] Example 13: Synthesis of Compound 8

[0112] The synthetic route of compound 8 is as follows:

[0113]

[0114] The specific synthesis steps are as follows:

[0115] The synthesis method refers to Examples 5 and 7, the raw material is replaced by 1-Boc-4-aminopiperidine with 1-Boc-4-aminomethylpiperidine, and 35 mg of white solid is obtained. UPLC-MS calculated for C 50 H 49 F4N7O7S[M+H] + :968.34,found:968.48. 1H NMR (400MHz, DMSO-d6) δ9.82(s,2H),8.90(t,J=5.8Hz,1H),8.54(t,J=5.7Hz,1H),8.41(d,J=8.3Hz,1H),8.32(s,1H),8.30 (d,J=1.9Hz,1H),8.08(dd,J=8.3,1.9Hz,1H),7.79(t,J=8.0Hz,1H),7.44(dd,J=10.6,1.9Hz,1H),7.34(dd,J=8.1,1.9Hz,1 H),7.21(q,J=8.0Hz,4H),6.72(s,1H),6.44(s,1H),4.41(d,J=13.0Hz,2H),3.52(t,J=4.5Hz,4H),3.35(s,4H),3.02–2.92 (m,4H),2.30(t,J=4.4Hz,4H),2.10–2.00(m,1H),1.95–1.84(m,2H),1.70–1.58(m,2H),1.54(s,6H),0.90(d,J=6.9Hz,6H).

[0116] Example 14: Synthesis of Compound 9

[0117] The synthetic route of compound 9 is as follows:

[0118]

[0119] The specific synthesis steps are as follows:

[0120] The synthesis method was based on Examples 7 and 10 to obtain 22 mg of a white solid. UPLC-MS calculated for C 55 H 59 F4N9O6S[M+H] + :1050.42,found:1050.62. 1H NMR (400MHz, DMSO-d6) δ9.70(s,1H),9.68(s,1H),8.90(t,J=5.8Hz,1H),8.54(t,J=5.7Hz,1H),8.41(d,J=8.3Hz,1H),8.30(d,J=1.9Hz ,1H),8.08(dd,J=8.3,1.9Hz,1H),7.79(t,J=8.0Hz,1H),7.44(dd,J=10.6,1.9Hz,1H),7.34(dd,J=8.1,1.9Hz,1H),7.21(q,J=8.0Hz,4H ),6.72(s,1H),6.44(s,1H),4.44(d,J=12.8Hz,2H),3.59(s,2H),3.55(t,J=4.6Hz,4H),3.40(s,2H),3.35(s,8H),3.12–2.98(m,4H),2 .96(p,J=6.9Hz,1H),2.78(s,3H),2.31(t,J=4.4Hz,4H),2.10–2.00(m,1H),1.78(d,J=11.5Hz,2H),1.56(s,6H),0.99(d,J=6.9Hz,6H).

[0121] Example 15: Synthesis of Compound 10

[0122] The synthetic route of compound 10 is as follows:

[0123]

[0124] The specific synthesis steps are as follows:

[0125] Step 1: Add S34 (150 mg, 0.33 mmol), DIEA (86 mg, 0.66 mmol) and DCM (10 mL) to a reaction flask, replace the argon atmosphere three times, add HATU (189 mg, 0.50 mmol) and stir at room temperature for 1 h, add mono-Boc propylenediamine (63 mg, 0.36 mmol) and stir at room temperature overnight, add water (50 mL), extract with ethyl acetate (50 mL*2), wash the organic phase with saturated brine (50 mL), dry over anhydrous sodium sulfate, filter and concentrate to obtain a crude product, and obtain 200 mg of light yellow solid S50 by column chromatography (DCM: MeOH = 20: 1), with a yield of 85%. UPLC-MS calculated for C 28 H 29 F4N5O4S[M+H] + :608.19,found:608.62.

[0126] Step 2: Add intermediate S50 (200 mg, 0.31 mmol) to the reaction flask, add HCl (0.77 mL, 3.1 mmol, 4M 1,4-dioxane solution) dropwise at room temperature, and stir for 1 h. Add 2 mL of methanol, filter, concentrate, and pump dry to obtain 210 mg of white solid, with a yield of 99%. UPLC-MS calculated for C 23 H 21 F4N5O2S[M+H] + :508.14,found:508.21.

[0127] Step 3: S8 (1 g, 1.55 mmol) was dissolved in THF (10 mL), and a solution of LiOH (111 mg, 4.64 mmol) in water (2 mL) was added dropwise at room temperature and stirred for 3 h. The pH was adjusted to 3 with 1 M HCl, and the aqueous phase was directly freeze-dried to obtain a crude product, which was dissolved in acetonitrile (20 mL), filtered and concentrated to obtain 766 mg of solid S52, with a yield of 85%. UPLC-MS calculated for C 38 H 38 N2O6[MH] - :617.24,found:617.73.

[0128] Step 4: S52 (100 mg, 0.16 mmol) and DCM (5 mL) were added to the reaction flask, argon was replaced three times, SOCl2 (100 mg, 0.32 mmol) and 1 drop of DMF were added, stirred at room temperature for 2 h, concentrated to obtain the acid chloride, S51 (82 mg, 0.16 mmol), triethylamine (50 mg, 0.48 mmol) and DCM (3 mL) were added to the reaction flask, the obtained acid chloride was dissolved in 2 mL of DCM, slowly added dropwise, stirred at room temperature for 2 h, the reaction solution was directly concentrated, and the oil pump was pulled dry to obtain 200 mg of light yellow solid, with a yield of 100%. UPLC-MS calculated for C 61 H 57 F4N7O7S[M+H] + :1108.40,found:M / 2+1=554.81.

[0129] Step 5: S53 (200 mg) was dissolved in 10 mL DCM, argon was replaced three times, BCl3 (2 mL, 1 M toluene solution) was added dropwise in an ice-water bath, stirred for 2 h in an ice-water bath, and NaHCO3 was added to neutralize to pH = 7, acetonitrile was added to dissolve, and the crude product was filtered and concentrated. 56 mg of white solid was obtained by reverse phase preparative purification, with a yield of 33%. UPLC-MS calculated for C 47 H 45 F4N7O7S[M+H]+ :928.30,found:928.41. 1 H NMR (400MHz, DMSO-d6) δ9.76(s,2H),8.90(t,J=5.8Hz,1H),8.54(t,J=5.7Hz,1H),8.41(d,J=8.3Hz,1H),8.30(d ,J=1.9Hz,1H),8.08(dd,J=8.3,1.9Hz,1H),7.79(t,J=8.0Hz,1H),7.44(dd,J=10.6,1.9Hz,1H),7.34(dd,J=8.1 ,1.9Hz,1H),7.21(q,J=8.0Hz,4H),6.72(s,1H),6.44(s,1H),3.54(t,J=4.5Hz,4H),3.40(s,2H),3.29(q,J=6.6 Hz, 4H), 2.96 (p, J = 6.9Hz, 1H), 2.31 (t, J = 4.4Hz, 4H), 1.73 (p, J = 6.8Hz, 2H), 1.54 (s, 6H), 0.89 (d, J = 6.9Hz, 6H).

[0130] Example 16: Synthesis of Compound 11

[0131] The synthetic route of compound 11 is as follows:

[0132]

[0133] The specific synthesis steps are as follows:

[0134] The synthesis method was based on Example 15 to obtain 32 mg of a white solid. UPLC-MS calculated for C 48 H 47 F4N7O7S[M+H] + :941.32,found:941.86. 1H NMR (400MHz, DMSO-d6) δ9.81(s,1H),9.70(s,1H),8.85(t,J=5.7Hz,1H),8.53(t,J=5.7Hz,1H),8.41(d,J=8. 3Hz,1H),8.30(s,1H),8.08(d,J=8.3Hz,1H),7.76(t,J=8.0Hz,1H),7.42(d,J=10.5Hz,1H),7.32(d,J=8.2Hz ,1H),7.25–7.17(m,4H),6.72(s,1H),6.44(s,1H),3.54(t,J=4.5Hz,4H),3.40(s,2H),3.21(dt,J=13.0,6.5 Hz, 4H), 2.96 (p, J = 6.9Hz, 1H), 2.32 (t, J = 4.5Hz, 4H), 1.54 (s, 6H), 1.37–1.29 (m, 2H), 0.89 (d, J = 6.8Hz, 6H).

[0135] Example 17: Synthesis of Compound 12

[0136] The synthetic route of compound 12 is as follows:

[0137]

[0138] The specific synthesis steps are as follows:

[0139] The synthesis method was based on Example 15 to obtain 40 mg of a white solid. UPLC-MS calculated for C 49 H 49 F4N7O7S[M+H] + :956.34,found:956.42. 1H NMR (400MHz, DMSO-d6) δ9.81(s,1H),9.70(s,1H),8.85(t,J=5.7Hz,1H),8.53(t,J=5.7Hz,1H),8.41(d,J=8.3 Hz,1H),8.30(s,1H),8.08(d,J=8.3Hz,1H),7.76(t,J=8.0Hz,1H),7.42(d,J=10.5Hz,1H),7.32(d,J=8.2Hz,1H ),7.25–7.17(m,4H),6.72(s,1H),6.44(s,1H),3.54(t,J=4.5Hz,4H),3.40(s,2H),3.21(dt,J=13.0,6.5Hz,4 H), 2.96 (p, J = 6.9Hz, 1H), 2.32 (t, J = 4.5Hz, 4H), 1.53–1.45 (m, 10H), 1.37–1.29 (m, 2H), 0.88 (d, J = 6.8Hz, 6H).

[0140] Example 18: Synthesis of Compound 13

[0141] The synthetic route of compound 13 is as follows:

[0142]

[0143] The specific synthesis steps are as follows:

[0144] The synthesis method was based on Example 15 to obtain 25 mg of a white solid. UPLC-MS calculated for C 50 H 51 F4N7O7S[M+H] + :970.35,found:970.62. 1H NMR (400MHz, DMSO-d6) δ9.80(s,1H),9.73(s,1H),8.85(t,J=5.7Hz,1H),8.53(t,J=5.7Hz,1H),8.41(d,J=8.3 Hz,1H),8.30(s,1H),8.08(d,J=8.3Hz,1H),7.76(t,J=8.0Hz,1H),7.42(d,J=10.5Hz,1H),7.32(d,J=8.2Hz,1H ),7.25–7.17(m,4H),6.72(s,1H),6.44(s,1H),3.54(t,J=4.5Hz,4H),3.40(s,2H),3.21(dt,J=13.0,6.5Hz,4 H), 2.96 (p, J = 6.9Hz, 1H), 2.32 (t, J = 4.5Hz, 4H), 1.53–1.45 (m, 10H), 1.37–1.29 (m, 4H), 0.89 (d, J = 6.8Hz, 6H).

[0145] Example 19: Synthesis of Compound 14

[0146] The synthetic route of compound 14 is as follows:

[0147]

[0148] The specific synthesis steps are as follows:

[0149] The synthesis method was based on Example 15 to obtain 32 mg of a white solid. UPLC-MS calculated for C 47 H 44 F4N8O7S[M+H] + :941.30,found:941.97. 1H NMR(400MHz, DMSO-d6)δ9.76(s,2H),8.90(t,J=5.8Hz,1H),8.54(t,J=5.7Hz,1H),8.50(s,1H),8.41(d,J= 8.3Hz,1H),8.30(d,J=1.9Hz,1H),8.08(dd,J=8.3,1.9Hz,1H),7.79(t,J=8.0Hz,1H),7.21(q,J=8.0Hz,4H) ,6.72(s,1H),6.44(s,1H),3.54(t,J=4.5Hz,4H),3.40(s,2H),3.29(q,J=6.6Hz,4H),2.96(p,J=6.9Hz,1H) ,2.38–6(m,4H),2.31(t,J=4.4Hz,4H),1.73(p,J=6.8Hz,2H),1.78(p,J=7.1Hz,2H),0.87(d,J=6.9Hz,6H).

[0150] Example 20: Synthesis of Compound 15

[0151] The synthetic route of compound 15 is as follows:

[0152]

[0153] The specific synthesis steps are as follows:

[0154] The intermediate S16 (100 mg, 0.23 mmol), S51 (186 mg, 0.34 mmol), HOBt (46 mg, 0.34 mmol), DIEA (0.12 mL, 0.69 mmol) and DMF (10 mL) were added to the reaction bottle, and PyBOP (177 mg, 0.34 mmol) was added. The argon gas was replaced three times, and the mixture was stirred at room temperature overnight. The mixture was purified by reverse preparative liquid phase to obtain 77 mg of a white solid with a yield of 36%. UPLC-MS calculated for C 47 H 46 F4N8O6S[M+H] + :927.32,found:927.98. 1H NMR (400MHz, DMSO-d6) δ11.91(s,1H), δ9.81(s,1H),9.70(s,1H),8.90(t,J=5.8Hz,1H),8.54(t,J=5.7Hz,1H),8.41(d, J=8.3Hz,1H),8.30(d,J=1.9Hz,1H),8.08(dd,J=8.3,1.9Hz,1H),7.79(t,J=8.0Hz,1H),7.44(dd,J=10.6,1.9Hz,1H),7 .34(dd,J=8.1,1.9Hz,1H),7.21(q,J=8.0Hz,4H),6.72(s,1H),6.44(s,1H),3.54(t,J=4.5Hz,5H),3.40(s,2H),3.29(q ,J=6.6Hz,4H),2.96(p,J=6.9Hz,1H),2.31(t,J=4.4Hz,4H),1.73(p,J=6.8Hz,2H),1.54(s,6H),0.89(d,J=6.9Hz,6H).

[0155] Example 21: Synthesis of Compound 16

[0156] The synthetic route of compound 16 is as follows:

[0157]

[0158] The specific synthesis steps are as follows:

[0159] Step 1: Add S25 (60 mg, 0.13 mmol) and 1,4-dioxane to a reaction flask, add a 20% aqueous solution of NaOH (10 mg, 0.25 mmol), stir overnight at room temperature, add 20 mL of water, neutralize with 1 M HCl to pH = 5, extract with ethyl acetate (50 mL*2), wash the organic phase with saturated brine (50 mL), dry over anhydrous Na2SO4, filter and concentrate to obtain 38 mg of light yellow solid S66. UPLC-MS calculated for C 23 H 26 N4O5[MH]-:437.19,found:437.48.

[0160] Step 2: Add intermediate S66 (35 mg, 0.08 mmol), S51 (65 mg, 0.12 mmol), HOBt (16 mg, 0.12 mmol), DIEA (0.06 mL, 0.36 mmol) and DMF (10 mL) to a reaction flask, add PyBOP (62 mg, 0.12 mmol), replace the argon gas three times, stir at room temperature overnight, and purify by reverse preparative liquid phase to obtain 16 mg of a white solid with a yield of 22%. UPLC-MS calculated for C 46 H 45 F4N9O6S[M+H] + :928.32,found:928.96. 1 H NMR (400MHz, DMSO-d6) δ9.74(s,2H),8.88(t,J=5.8Hz,1H),8.53(t,J=5.7Hz,1H),8.40(d,J=8.3Hz,1H),8.31(d ,J=1.9Hz,1H),8.07(dd,J=8.3,1.9Hz,1H),7.77(t,J=8.0Hz,1H),7.42(dd,J=10.6,1.9Hz,1H),7.31(dd,J=8.1 ,1.9Hz,1H),7.20(q,J=8.0Hz,4H),6.72(s,1H),6.44(s,1H),3.54(t,J=4.5Hz,5H),3.40(s,2H),3.29(q,J=6.6 Hz, 4H), 2.96 (p, J = 6.9Hz, 1H), 2.31 (t, J = 4.4Hz, 4H), 1.73 (p, J = 6.8Hz, 2H), 1.54 (s, 6H), 0.88 (d, J = 6.9Hz, 6H).

[0161] Example 22: Biological evaluation test

[0162] Test Example 1: Evaluation of the effect of reducing androgen receptor (AR) expression

[0163] AR-positive human prostate cancer cells VCaP and 22RV1 were cultured in RPMI 1640 (hereinafter referred to as evaluation medium) containing 10% FBS to a concentration of 3×10 5 / well was inoculated into a 6-well microplate (Corning) and cultured overnight. The evaluation medium containing the example compound was added to the culture to make the final concentration of the example compound reach 10μmol / L, and the culture was carried out for 24 hours. After 24 hours of culture, the culture medium was removed, and the cells were washed with PBS. Then, RIPA lysis buffer containing 1% Protease Inhibitor Cocktail was added. After lysis and centrifugation, a total protein extract was obtained, and the protein concentration in the extract was detected by the BCA method; SDS-PAGE was used for protein electrophoresis, and then the protein was transferred to a PVDF (MilliporeSigma IPVH00010) membrane by constant current electrophoresis at 200mA for 90min; the PVDF membrane was placed in a solution containing 5% skim milk and blocked at room temperature for 1h; the immunoreaction was carried out using the Anti-Androgen Receptor antibody [EPR1535(2)] (HRP) (abcam); after washing the membrane, ECL luminescent liquid was added and exposed. The grayscale analysis of the bands was performed using the software Image J. GAPDH protein band was detected for each sample as an internal reference, and the AR protein degradation rate of the example compound was calculated based on the grayscale of the protein band.

[0164] The results of VCaP and 22Rv1 cells are shown in Table 1. Some of the compounds in the examples showed a certain AR protein degradation effect. In the table, " / " means that the protein is not significantly down-regulated; "reduced" means that the AR protein expression is down-regulated by >50%).

[0165] Table 1

[0166]

[0167]

[0168] Test Example 2: Androgen-dependent prostate cancer cell proliferation inhibitory activity

[0169] Androgen receptor-positive human prostate cancer cells 22RV1 and VCaP were cultured in RPMI 1640 medium containing 5% carbon-stripped serum (CCS) (hereinafter referred to as evaluation medium) at a concentration of 4×10 3 The amount of 100 μg / well was inoculated into a transparent bottom 96-well microplate (Corning) and cultured for 48 h. To the culture, an evaluation medium containing R1881 (final concentration of R1881 was 0.1 nmol / L) and an evaluation medium containing the example compound were added.

[0170] The final concentrations of the example compounds were 10, 100, and 1000 nmol / L. After 5 days of culture, the number of viable cells was determined using WST-1 (Roche). The cell proliferation activity value of 0.1 nmol / L R1881 was set to 100%, and the cell proliferation activity of only the evaluation culture medium was set to 0%, and the anti-tumor proliferation activity of the example compounds was preliminarily evaluated.

[0171] The results of cell proliferation inhibition are shown in Table 2. The example compounds showed certain prostate cancer cell proliferation inhibition activity (“++” indicates the estimated IC 50 10-100nM, "+" indicates estimated IC 50 100-1000nM, "NA" indicates estimated IC 50 Greater than 1000nM,).

[0172] Table 2

[0173]

[0174] Some example compounds (compounds 1, 2, 3 and 10) were selected for further activity verification, with final concentrations of 1.53, 4.6, 13.8, 41.1, 123.4, 370.3, 1111, 3333 and 10000 nmol / L. After 5 days of culture, the number of viable cells was determined. The number of viable cells was determined using WST-1 (Roche). The cell proliferation activity value of 0.1 nmol / L R1881 was set to 100%, and the cell proliferation activity of only the evaluation medium was set to 0%. The 50% proliferation inhibition concentration (IC 50 value). The result is Figure 1 and Figure 2 shown.

Claims

1. A compound as shown in formula I, its optical isomers, isotopic derivatives or pharmaceutically acceptable salts thereof, In Formula I, Ring A is selected from C5-C10 heteroaryl; Ring B is selected from C 4-10 C ring is selected from C6-C10 membered aryl, C5-C10 membered heteroaryl; X, Y, Z are C or N; R 1 , R 2 , R 3 They are independently selected from H, halogen, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl, OH, NH2, CN, nitro, carboxyl, and C3-6 cycloalkyl; and the C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl, OH, NH2, carboxyl, and C3-6 cycloalkyl may contain 1, 2 or 3 substituents selected from halogen, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl, OH, NH2, CN, nitro, and carboxyl, respectively. R 4 Selected from H, halogen, C1-C6 alkyl, C1-C6 haloalkyl; R 5 , R 6 are each independently selected from H, halogen, C1-C6 alkyl, C1-C6 haloalkyl, and R 3 and R 4 Connected to form a ring with the carbon atom shared by both; R 7 , R 8 Selected from H, halogen, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl, OH, NH2, CN, nitro, carboxyl, C3-C6 cycloalkyl; L 1 , L 2 , L 3 , L 4 and L 5 Each is independently and compatibly selected from a single bond, a C1-C6 alkylene group, a carbonyl group, a 4-10 membered heterocycloalkylene group, -O-, -N(R B1 )R B2 -, where R B1 is H, C1-C6 alkyl, R B2 It is a C0-C6 alkylene group.

2. The compound according to claim 1, its optical isomer, isotope derivative or pharmaceutically acceptable salt thereof, characterized in that: In Formula I, Ring A is selected from Ring B is selected from The C ring is selected from X is C or N; Y is C; Z is C; R 1 Selected from -OH; R 2 Selected from -OH; R 3 Selected from R 4 Selected from -CF3; R 5 Selected from -CH3 or -CH2CH2-, one side of which is connected to R 6 Connect; R 6 Selected from -CH3 or -CH2CH2-, one side of which is connected to R 5 Connect; R 7 Selected from -F;R 8 Selected from -H; Selected from 3. The compound according to claim 1, its optical isomer, isotope derivative or pharmaceutically acceptable salt thereof, characterized in that: It is any one of the following structural formulas: Compound 1: Compound 2: Compound 3: Compound 4: Compound 5: Compound 6: Compound 7: Compound 8: Compound 9: Compound 10: Compound 11: Compound 12: Compound 13: Compound 14: Compound 15: Compound 16:

4. A pharmaceutical composition comprising the compound according to any one of claims 1 to 3, its optical isomer, isotopic derivative or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

5. Use of the compound according to any one of claims 1 to 3, its optical isomer, isotope derivative or pharmaceutically acceptable salt thereof in the preparation of a drug for inducing degradation or decrease of androgen receptor.

6. Use of the compound according to any one of claims 1 to 3, its optical isomers, isotopic derivatives or pharmaceutically acceptable salts thereof in the preparation of a medicament for preventing or treating prostate cancer.

7. The method for preparing the compound according to any one of claims 1 to 3, characterized in that: The synthetic route of compound 1 is as follows: The synthetic route of compound 2 is as follows: The synthetic route of compound 3 is as follows: The synthetic route of compound 4 is as follows: The synthetic route of compound 5 is as follows: The synthetic route of compound 6 is as follows: The synthetic route of compound 7 is as follows: The synthetic route of compound 8 is as follows: The synthetic route of compound 9 is as follows: The synthetic route of compound 10 is as follows: The synthetic route of compound 11 is as follows: The synthetic route of compound 12 is as follows: The synthetic route of compound 13 is as follows: The synthetic route of compound 14 is as follows: The synthetic route of compound 15 is as follows: The synthetic route of compound 16 is as follows:

8. The method for preparing the compound according to claim 7, characterized in that: The synthetic route of intermediate S10 is as follows: The synthetic route of intermediate S16 is as follows: The synthetic route of intermediate S19 is as follows: The synthetic route of intermediate S25 is as follows: The synthetic route of intermediate S30 is as follows:

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