Ubiquitin specific protease 28 small-molecule inhibitor as well as preparation method and application thereof

Abstract

The invention belongs to the field of medicinal chemistry, and discloses a ubiquitin specific protease 28 (USP28) small-molecule inhibitor, a preparation method thereof and application of the USP28 small-molecule inhibitor in medicine preparation. Relates to a compound as shown in a general formula I or pharmaceutically acceptable salt, solvate, hydrate, active metabolite, polymorphic substance, ester, isotope compound, stereoisomer or prodrug thereof, a pharmaceutical composition containing the compound with the structure as shown in the formula I and application of the pharmaceutical composition serving as a USP28 inhibitor to preparation of drugs for diseases including but not limited to cancers and the like. And # imgabs0 #.

Description

Technical Field

[0001] The present invention belongs to the field of pharmaceutical chemistry, and in particular relates to a ubiquitin-specific protease 28 (USP28) small molecule inhibitor, a preparation method thereof, and use thereof in preparing medicines. Background Art

[0002] The USP protein family, also known as the Ubiquitin-Specific Protease (USP) family, is a large number of deubiquitinating enzymes with diverse structures, which participate in a variety of biological functions. For example, regulating protein stability: by removing the ubiquitin chain on the protein, the protein is prevented from being degraded by the proteasome, thereby increasing the stability of the protein in the cell; for example, in cell cycle regulation, USP protein can stabilize some key cell cycle regulatory proteins to ensure the normal progress of the cell cycle. Participating in cell signal transduction: It can regulate the activity and stability of signal molecules and affect the signal transduction pathway in the cell; for example, in the Wnt signaling pathway, USP protein can stabilize β-catenin protein through deubiquitination, thereby enhancing Wnt signal transduction and promoting cell proliferation and differentiation. Maintaining ubiquitin homeostasis: It is of great significance to maintain the level of free ubiquitin in the cell; it can recycle and reuse ubiquitin molecules to ensure the normal operation of the ubiquitin system in the cell, and indirectly affect many cellular processes that rely on ubiquitination modification.

[0003] USP28 plays an important role in a variety of cancers and muscle-related diseases. USP28 is highly expressed in many human tumors, including head and neck cancer, gastric cancer, liver cancer, ovarian cancer, endometrial cancer, prostate cancer, thyroid cancer, renal cancer, lymphoma, neuroblastoma, and osteosarcoma. On the one hand, it can stabilize some oncogene proteins, such as c-Myc, to promote the proliferation and survival of cancer cells. USP28 also promotes tumor suppressor pathways, such as by deubiquitinating TP53, a function that is offset by nuclear caspase 8 during tumor recurrence. USP28 also regulates the DNA damage response in cancer cells by deubiquitinating and stabilizing checkpoint kinase 2 (HK2), TP53 protein 1 (TP53BP1), and claspn (CLSPN), thereby preventing apoptosis and establishing cell cycle arrest to promote DNA repair. The exact function is still under study. On the other hand, high expression of USP28 is associated with poor prognosis of tumors. USP28 has become a potential target for cancer therapy. In addition, high expression of USP28 increases cardiac hypertrophic cardiomyocytes, which are mainly distributed in cardiomyocytes. It negatively regulates antioxidant responses by deubiquitinating and stabilizing TRIM21, increasing oxidative stress in cardiomyocytes and promoting cardiac hypertrophy and damage. Therefore, USP28 inhibitors are expected to become effective drugs with new mechanisms of action for the treatment of cancer and muscle-related diseases.

[0004] The research on USP28 inhibitors has become a hot topic in the development of new drugs, and the research on enzyme structure and crystal structure is increasing. Up to now, although many USP28 inhibitors have been discovered or developed, including some highly active compounds, most of these molecules have poor selectivity, and there is still a lack of effective selective inhibitors against USP28 targets in clinical practice. Therefore, more effective USP28 selective inhibitors are needed, and their development potential is still huge. Summary of the invention

[0005] Aiming at the USP28 target, the present invention aims to provide compounds with good activity and can be used as USP28 inhibitors, which can be used to prevent or treat diseases related to excessive USP28 activity in humans or mammals.

[0006] To achieve the purpose of the present invention, the USP28 inhibitor of the present invention is a compound as shown in the general formula I or a pharmaceutically acceptable salt, solvate, hydrate, active metabolite, polymorph, ester, isotope compound, stereoisomer or prodrug thereof.

[0007] Where: The nitrogen-containing A ring represented by the dotted line is selected from one of the following substituents: X is selected from one of the following substituents: R1 is selected from the following substituents: hydrogen, or R2 is selected from the following substituents: hydrogen, or

[0008] The substituents of the ubiquitin-specific protease 28 small molecule inhibitor are preferably as follows:

[0009] Ring A is selected from one of the following substituents:

[0010] X is selected from one of the following substituents:

[0011] R1 is selected from the following substitutions:

[0012] R2 is selected from the following substituents:

[0013] The ubiquitin-specific protease 28 small molecule inhibitor is characterized in that compound 19 is preferred. The use of any of the above compounds in the preparation of drugs for preventing or treating diseases mediated by USP28, including but not limited to cancer and muscle-related diseases. For example, it can be applied to the preparation of drugs for Fanconi anemia, muscle atrophy or tumor vascular disease.

[0014] The cancer includes bladder cancer, pancreatic cancer, breast cancer, colon cancer, lung cancer, glioma, melanoma, head and neck cancer, gastric cancer, liver cancer, ovarian cancer, endometrial cancer, prostate cancer, thyroid cancer or kidney cancer.

[0015] The preparation route of the small molecule inhibitor is as follows, taking one of the A ring substituents as an example: The specific steps are as follows; (a) In a solvent, 5-chloroindole (II) is used as a starting material and formylated in the presence of phosphorus oxychloride and N,N-dimethylformamide to obtain intermediate III; the solvent used is N,N-dimethylformamide. (b) Add a basic compound to a solvent, and under basic conditions, the formylated intermediate III obtained in step a) undergoes a substitution reaction with benzyl chloride or sulfonyl chloride to obtain intermediate IV. The solvent used is tetrahydrofuran, ethanol or dichloromethane, and the basic compound used is N,N-diisopropylethylamine or sodium hydride. (c) In tetrahydrofuran solution, the intermediate IV obtained in step b) reacts with bromomethylbutenolide in the presence of zinc powder and saturated ammonium chloride to obtain compound I. Each product obtained in the above reaction route can be obtained by conventional separation techniques, including but not limited to filtration, distillation, crystallization, chromatography, etc. The starting materials required for the synthesis can be synthesized by oneself or purchased from commercial institutions. The compounds described in the present invention can be synthesized using synthetic methods to obtain a single optical isomer or a mixture of optical isomers. Advantages of the present invention: The synthetic route is simple. The synthesized compounds show good inhibitory activity against USP28 enzyme through biological activity evaluation, especially compound 19, whose single-point 25 μM initial screening inhibition rate IC 50 The concentration of the USP28 inhibitor is 1.772 μM. The active ingredient is used as a USP28 inhibitor to prepare drugs including but not limited to cancer and muscle-related diseases. DETAILED DESCRIPTION

[0016] The present invention can be better understood according to the following examples. However, it is easy for those skilled in the art to understand that the contents described in the embodiments are only used to illustrate the present invention, and should not and will not limit the present invention. Unless otherwise specified, the percentage contents are all by mass. Example 1 Synthesis of Compound 1

[0017] Intermediate 1-1 1-p-Toluenesulfonyl-1H-pyrrole-3-carbaldehyde

[0018] To a solution of pyrrole-3-carboxaldehyde (200 mg, 2.10 mmol, 1 eq) in anhydrous tetrahydrofuran (5 mL) was slowly added 60% sodium hydride (126 mg, 3.15 mmol, 1.5 eq) under ice bath, and then 4-methylbenzenesulfonyl chloride (600 mg, 3.15 mmol, 1.5eq) and reacted under ice bath. After the reaction was completed as monitored by thin layer chromatography, tetrahydrofuran was removed in vacuo, the mixture was dissolved in ethyl acetate, washed with brine (5mL×3), the organic phase was dried over anhydrous sodium sulfate, the solvent was removed in vacuo, and the intermediate 1-toluenesulfonyl-1H-pyrrole-3- formaldehyde. 1H NMR (400MHz, DMSO-d6) δ9.79 (s, 1H), 8.32 (t, J = 2.4Hz, 1H), 8.15-8.11 (m, 1H), 8.11-8.09 (m, 1H),7.82-7.79(m,1H),7.79-7.77(m,1H),7.56(t,J=2.4Hz,1H),6.71(dd,J=3.3,1.4Hz,1H).

[0019] Compound 1 4-(Hydroxy(1-toluenesulfonyl-1H-pyrrol-3-yl)methyl)-3-methylenedihydrofuran-2(3H)-one

[0020] Zinc powder (121 mg, 1.85 mmol, 5 eq) and bromomethylbutenolide (100 mg, 0.37 mmol, 1 eq) were added to a solution of 1-p-toluenesulfonyl-1H-pyrrole-3-carboxaldehyde (53 mg, 0.56 mmol, 1 eq) in anhydrous tetrahydrofuran (2 mL). After 10 min, 600 μL of saturated ammonium chloride solution was added and the mixture was stirred at room temperature for reaction. After the reaction was completed by TLC monitoring, the mixture was filtered and concentrated. Ethyl acetate and water (5 mL) were added to obtain the product. ×3) for three times, collect the organic phase, dry the organic phase over anhydrous sodium sulfate, and concentrate to obtain a crude product. Column chromatography purification afforded compound 1 as a white solid. 1 H NMR (400MHz, CDCl3) δ7.79(d,J=1.7Hz,1H),7.77(d,J=1.8Hz,1H),7.52(d,J=1 .8Hz,1H),7.50(d,J=1.7Hz,1H),7.14(dd,J=5.7,2.4Hz,2H),6.31(d,J=2.2Hz, 1H),6.28(dd,J=3.1,1.7Hz,1H),5.65(d,J=1.8Hz,1H),4.71(d,J=6.7Hz,1H),4.25 (dd,J=9.6,8.2Hz,1H),4.12(dd,J=9.6,3.9Hz,1H),3.34(m,J=10.4,4.1,2.1Hz,1H). 13 C NMR (101MHz, CDCl3) δ170.75,141.05,137.06,134.65,130.03,129.59,128.36,125.28,122.00, 118.43,112.30,69.81,67.64,44.75.

[0021] Example 2 Synthesis of Compound 2 4-((1-((4-bromophenyl)sulfonyl)-1H-pyrrol-3-yl)(hydroxy)methyl)-3-methylenedihydrofuran-2(3H)-one

[0022] The synthesis process is the same as in Example 1, except that 4-methylbenzenesulfonyl chloride is replaced by 4-bromobenzenesulfonyl chloride to obtain a white solid. 1 HNMR(400MHz, CDCl3) δ7.79(d,J=1.7Hz,1H),7.77(d,J=1.8Hz,1H),7.52(d,J=1.8 Hz,1H),7.50(d,J=1.7Hz,1H),7.14(dd,J=5.7,2.4Hz,2H),6.31(d,J=2.2Hz,1H), 6.28(dd,J=3.1,1.7Hz,1H),5.65(d,J=1.8Hz,1H),4.71(d,J=6.7Hz,1H),4.25(dd ,J=9.6,8.2Hz,1H),4.12(dd,J=9.6,3.9Hz,1H),3.34(m,J=10.4,4.1,2.1Hz,1H). 13 C NMR (101MHz, CDCl3) δ170.75,141.05,137.06,134.65,130.03,129.59,128.36,125.28,122.00,118.43,112.30,69.81,67.64,44.75.

[0023] Example 3 Synthesis of Compound 3 4-((1-((4-chlorophenyl)sulfonyl)-1H-pyrrol-3-yl)(hydroxy)methyl)-3-methylenedihydrofuran-2(3H)-one

[0024] The synthesis process is the same as in Example 1, except that 4-methylbenzenesulfonyl chloride is replaced by 4-chlorobenzenesulfonyl chloride to obtain a white solid. 1HNMR(400MHz,DMSO-d6)δ7.81(s,1H),7.79(s,1H),7.45(s,1H),7.43(s,1H),7 .30-7.27(t,1H),7.14(s,1H),6.31(dd,J=3.1,1.4Hz,1H),5.95(d,J=1.8Hz,1 H),5.58(s,1H),5.26(d,J=0.9Hz,1H),4.65(d,J=5.4Hz,1H),4.26(t,J=8.7Hz ,1H),4.11(dd,J=9.2,3.9Hz,1H),3.33(m,J=7.9,5.7,3.7Hz,1H),2.38(s,3H). 13 C NMR (101MHz, DMSO-d6) δ170.37,145.33,135.31,135.24,130.77,130.22,126.64,123.12,121.51,118.16,112.87,68.45,67.66,43.91,21.06. Example 4 Synthesis of Compound 4 4-((5-Chloro-1-(2-fluorobenzyl)-1H-indol-3-yl)(hydroxy)methyl)-3-methylenedihydrofuran-2(3H)-one

[0025] Intermediate 4-1 5-Chloro-1H-indole-3-carbaldehyde

[0026] Under ice bath condition, DMF (0.3mL, 4.02mmol, 2eq) was taken into the reaction bottle, POCl3 (0.37mL, 4.20mmol, 2eq) was added dropwise during stirring, and the mixture was stirred until it reached a brown-red viscous state. 5-Chloroindole (300mg, 2.00mmol, 1eq) dissolved in DMF was slowly added, and the mixture was stirred for about 30 minutes. The reaction system gradually became a semi-solid state. Stirring was stopped, and about 100ml of ice water was added. The system was dissolved and changed from a semi-solid state to a dark red liquid state. The pH was adjusted to 7-8 with 10% NaOH solution under ice bath. A large amount of white solid was produced in the system. The mixture was stirred overnight, and the solid was filtered off by suction. After drying, the solid was recrystallized with CH2Cl2. After filtering off by suction, 5-chloro-1-hydrogen-indole-3-aldehyde was obtained as a brown-yellow solid powder with a yield of 99%. 1H NMR (400MHz, DMSO-d6) δ12.31(s,1H),9.94(s,1H),8.37(d,J=3.1Hz,1H),8.07(d,J=2.0Hz,1H),7.55(d,J=8.6Hz,1H),7.29(dd,J=8.6,2.1Hz,1H).

[0027] Intermediate 4-2 5-Chloro-1-(2-fluorophenyl)-1H-indole-3-carbaldehyde

[0028] Under ice bath conditions, 5-chloro-1-hydrogen-indole-3-carboxaldehyde (901 mg, 5.02 mmol, 1 eq) and sodium hydride (367 mg, 15.31 mmol, 3 eq) were put into a reaction bottle, 4 ml of tetrahydrofuran was added to dissolve, and 2-fluoro-benzyl chloride (1.45 g, 10.03 mmol, 2 eq) was added after stirring at room temperature for about 30 min. The mixture was stirred at room temperature and the reaction was complete after monitoring by thin layer chromatography. 3 ml of water was added to quench the reaction, and the mixture was filtered off with suction. The tetrahydrofuran was removed by rotary evaporation under reduced pressure, and the mixture was extracted with saturated brine and ethyl acetate. The organic phase was dried over anhydrous magnesium sulfate, and the filtrate was concentrated and subjected to column chromatography to obtain 5-chloro-1-(2-fluorophenyl)-1-hydrogen-indole-3-carboxaldehyde.

[0029] Compound 4 4-((5-Chloro-1-(2-fluorobenzyl)-1H-indol-3-yl)(hydroxy)methyl)-3-methylenedihydrofuran-2(3H)-one

[0030] Bromomethylbutenolide (552 mg, 3.12 mmol, 1.5 eq) was added to a reaction flask, and about 3 ml of tetrahydrofuran solution was used as a solvent. Zinc powder (666 mg, 10.18 mmol, 5 eq) was added and stirred at room temperature for 10 min. Then, intermediate 4-2 (381 mg, 2.13 mmol, 1 eq) was added and stirred at room temperature for 5 min. A saturated ammonium chloride solution was added dropwise and stirred at room temperature. The reaction was detected by thin layer chromatography until the reaction was complete. The reaction was filtered and the tetrahydrofuran was removed by vacuum rotary evaporation. The reaction was extracted with saturated brine and ethyl acetate. The organic phase was dried over anhydrous sodium sulfate. The filtrate was concentrated by suction and column chromatography to obtain compound 4 as a white solid with a yield of 54%. 1H NMR (400MHz, CDCl3) δ7.72(d,J=1.8Hz,1H),7.33-7.27(overlap,2H),7.20-7.17(m,1H),7.16(s,1H),7.14-7.03(m,2H),6.93(t,J=7.5Hz,1 H),6.38(d,J=2.1Hz,1H),5.89(s,1H),5.31(s,2H),4.93(d,J=8.1Hz,1H),4.25-4.19(m,1H),4.10-4.05(m,1H),3.62(brs,1H),2.20(s,1H). 13 C NMR (100MHz, CDCl3) δ170.65,160.49,135.47,135.23,130.11,129.08,127.62,126.86,126.06 ,125.46,124.63,123.52,123.14,119.16,115.80,115.01,111.19,69.94,68.09,44.60,44.35.

[0031] Example 5 Synthesis of Compound 5 4-((5-Chloro-1-(3-fluorobenzyl)-1H-indol-3-yl)(hydroxy)methyl)-3-methylenedihydrofuran-2(3H)-one

[0032] The synthesis process is the same as in Example 4, except that 2-fluoro-benzyl chloride is replaced with 3-fluorobenzyl chloride to obtain a light yellow solid with a yield of 52%. 1 H NMR (400MHz, CDCl3) δ7.73 (s, 1H), 7.33-7.27 (m, 1H), 7.18 (d, J = 1.0Hz, 2H) ,7.13(s,1H),7.01-6.96(m,1H),6.86(d,J=7.7Hz,1H),6.74(d,J=9.3Hz,1 H), 6.39 (d, J = 2.1Hz, 1H), 5.89 (d, J = 1.4Hz, 1H), 5.27 (s, 2H), 4.95 (d, J = 8. 0Hz,1H),4.30–4.19(m,1H),4.15-4.05(m,1H),3.64(brs,1H),2.22(s,1H). 13C NMR (100MHz, CDCl3) δ170.54,163.14,139.05,135.37,136.21,130.66,127.58,126.92,126.13 ,125.47,123.27,122.21,119.26,115.12,113.69,111.24,69.87,67.97,49.92,44.65,29.90.

[0033] Example 6 Synthesis of Compound 6 4-((5-Chloro-1-(2-chlorobenzyl)-1H-indol-3-yl)(hydroxy)methyl)-3-methylenedihydrofuran-2(3H)-one

[0034] The synthesis process is the same as in Example 4, except that 2-fluoro-benzyl chloride is replaced by 2-chlorobenzyl chloride to obtain a white solid with a yield of 53%. 1 HNMR(400MHz, CDCl3)δ7.74(s,1H),7.43(d,J=8.8Hz,1H),7.28–7.08(m,5H),6.68(d,J=7.6Hz,1H),6.39(s,1H),5 .91(s,1H),5.37(s,2H),4.95(d,J=8.1Hz,1H),4.33-4.18(m,1H),4.16-4.05(m,1H),3.66(brs,1H),2.23(s,1H). 13 C NMR (100MHz, CDCl3) δ170.54,135.45,135.25,133.96,132.80,129.88,129.44,128.25,127.67 ,127.39,126.84,126.15,125.46,123.20,119.31,115.11,111.31,70.05,67.85,48.04,44.71.

[0035] Example 7 Synthesis of Compound 7 4-((5-chloro-1-(4-chlorobenzyl)-1H-indol-3-yl)(hydroxy)methyl)-3-methylenedihydrofuran-2(3H)-one

[0036] The synthesis process is the same as in Example 4, except that 2-fluoro-benzyl chloride is replaced by 4-chlorobenzyl chloride to obtain a white solid with a yield of 66%. 1HNMR(400MHz, CDCl3)δ7.73(s,1H),7.29(d,J=8.4Hz,2H),7.17(s,2H),7.11(s,1H),7.01(d,J=8.4Hz,2H),6.40(s,1H) ,5.90(s,1H),5.24(s,2H),4.95(d,J=8.1Hz,1H),4.22(d,J=8.0Hz,1H),4.12-4.03(m,1H),3.62(brs,1H),2.13(s,1H). 13 C NMR (100MHz, CDCl3) δ170.25,135.27,134.82,129.23,128.10,127.58,126.92,126.15 ,125.40,123.26,119.27,115.75,115.09,114.26,111.35,70.08,67.94,49.21,44.14.

[0037] Example 8 Synthesis of Compound 8 4-((5-chloro-1-(4-bromobenzyl)-1H-indol-3-yl)(hydroxy)methyl)-3-methylenedihydrofuran-2(3H)-one

[0038] The synthesis process is the same as in Example 4, except that 2-fluoro-benzyl chloride is replaced by 4-bromobenzyl chloride to obtain a light yellow solid with a yield of 57%. 1 HNMR (400MHz, DMSO-d6) δ7.77(d,J=2.0Hz,1H),7.52(s,1H),7.50(s,2H),7.45(d,J=8.8Hz,1H),7.12(d,J=8.3Hz,3H),6.04(s,1 H),5.64(d,J=4.6Hz,1H),5.41(s,1H),5.40(s,2H),5,03(t,J=7.5Hz,1H),4.35(t,J=8.7Hz,1H),4.27-4.19(m,1H),3.53(s,1H). 13 CNMR(100MHz,DMSO-d6)δ171.10,137.89,136.14,135.11,131.91,129.65,129.19,127.6 8,124.31,123.79,121.87,121.00,119.44,116.51,112.34,68.83,68.69,48.85,44.55.

[0039] Example 9 Synthesis of Compound 9 4-((5-chloro-1-(2-methylbenzyl)-1H-indol-3-yl)(hydroxy)methyl)-3-methylenedihydrofuran-2(3H)-one

[0040] The synthesis process is the same as in Example 4, except that 2-fluoro-benzyl chloride is replaced by 2-methylbenzyl chloride to obtain a white solid with a yield of 63%. 1 HNMR(400MHz, CDCl3)δ7.66(s,1H),7.30-6.98(overlap,5H),6.90(s,1H),6.67(d,J=7.6Hz,1H),6.29(s,1H),5.84(s,1 H),5.15(s,2H),4.82(d,J=8.2Hz,1H),4.11(t,J=8.3Hz,1H),3.98-3.94(m,1H),3.54(s,1H),2.19(s,3H),2.15(s,1H). 13 C NMR (100MHz, CDCl3) δ170.65,135.85,135.60,135.33,133.98,130.78,128.32,127.51,127.32,12 6.82,126.57,125.97,125.39,123.02,119.16,114.85,111.24,69.92,67.85,48.46,44.62,19.05.

[0041] Example 10 Synthesis of Compound 10 4-((5-chloro-1-(4-methylbenzyl)-1H-indol-3-yl)(hydroxy)methyl)-3-methylenedihydrofuran-2(3H)-one

[0042] The synthesis process was the same as in Example 4, except that 2-fluoro-benzyl chloride was replaced with 4-methylbenzyl chloride to obtain a light yellow solid with a yield of 48.67%. 1H NMR(400MHz, CDCl3) δ7.70(d,J=4.0Hz,1H),7.24 -7.20(m,1H),7.16(d,J=1.9Hz,1H),7.14 -7.12(m,1H),7.10(d,J=6.4Hz,2H),6.98(d,J=7.9Hz,2H),6.36(s,1H),5.88(s,1H),5.19(s,2H),4. 89(d,J=8.0Hz,1H),4.20(t,J=7.9Hz,1H),4.07–4.01(m,1H),3.62(s,1H),2.32(s,3H),1.26(s,1H). 13 C NMR (100MHz, CDCl3) δ170.82,137.91,135.41,135.24,133.33,129.63,127.61,126.86,12 6.81,125.83,125.44,122.92,119.05,114.67,111.37,69.88,67.93,50.18,44.49,21.09.

[0043] Example 11 Synthesis of Compound 11 4-((5-Chloro-1-(2,5-difluorobenzyl)-1H-indol-3-yl)(hydroxy)methyl)-3-methylenedihydrofuran-2(3H)-one

[0044] The synthesis process was the same as in Example 4, except that 2-fluoro-benzyl chloride was replaced with 2,5-difluorobenzyl chloride to obtain a white solid with a yield of 46%. 1 H NMR (400MHz, CDCl3) δ7.75 (d, J = 1.7Hz, 1H), 7.28 (s, 1H), 7.27-7.21 (overlapz ,2H),7.18(s,1H),7.15-7.06(m,1H),7.02-6.96(m,1H),6.62-6.53(m,1H),6.4 1(d,J=2.0Hz,1H),5.90(d,J=1.4Hz,1H),5.33(d,J=16.7Hz,2H),4.98(d,J=7.9 Hz,1H),4.30-4.22(m,1H),4..15-4.08(m,1H),3.68-3.62(m,1H),2.29(s,1H).

[0045] Example 12 Synthesis of Compound 12 4'-((5-chloro-3-(hydroxy(4-methylene-5-oxotetrahydrofuran-3-yl)methyl)-1H-indol-1-yl)methyl)-[1,1'-biphenyl]-3-carbonitrile

[0046] The synthesis process was the same as in Example 4, except that 2-fluoro-benzyl chloride was replaced with 4'-(chloromethyl)-[1,1'-biphenyl]-3-cyano to obtain a white solid with a yield of 41.80%. 1 H NMR (400MHz, CDCl3) δ7.79-7.77(m,2H),7.69-7.64(m,1H),7.53(d,J=8.3Hz,2H),7.51–7.45(m,2H),7.29(s,1H),7.25-7.18(overla p,4H),6.42(s,1H),5.93(s,1H),5.37(s,2H),4.99(d,J=7.9Hz,1H),4.32–4.23(m,1H),4.16-4.12(m,1H),3.70(s,1H),2.33(s,1H). 13 C NMR (100MHz, CDCl3) δ170.74,144.69,137.96,137.14,135.48,135.27,133.80,132.97,129.97,129.42,127.89,127 .71,127.10,126.94,126.14,125.54,123.27,119.26,118.61,115.16,111.34,111.17,70.07,67.97,49.94,45.02.

[0047] Example 13 Synthesis of Compound 13 4-((5-Chloro-1-(methylsulfonyl)-1H-indol-3-yl)(hydroxy)methyl)-3-methylenedihydrofuran-2(3H)-one

[0048] Intermediate 13-1 5-Chloro-1-(methylsulfonyl)-1-hydrogen-indole-3-carbaldehyde

[0049] Under ice bath conditions, intermediate 4-1 (901 mg, 5.02 mmol) and N,N-diisopropylethylamine (1.75 mL, 10.07 mmol, 2 eq) were put into a reaction flask, 4 ml of tetrahydrofuran was added to dissolve, and methanesulfonyl chloride (10.03 mmol) was added after stirring at room temperature for about 30 min. The mixture was stirred at room temperature and the reaction was complete after TLC monitoring. 3 ml of water was added to quench the reaction, and the mixture was filtered off with suction. The tetrahydrofuran was removed by rotary evaporation under reduced pressure, and the mixture was extracted with saturated brine and ethyl acetate. The organic phase was dried over anhydrous magnesium sulfate, and the filtrate was concentrated and subjected to column chromatography to obtain 5-chloro-1-(methylsulfonyl)-1-hydrogen-indole-3-carbaldehyde.

[0050] Bromomethylbutenolide (552 mg, 3.12 mmol, 1.5 eq) was added to a reaction flask, and about 3 ml of tetrahydrofuran solution was used as a solvent. Zinc powder (666 mg, 10.18 mmol, 5 eq) was added and stirred at room temperature for 10 min. Then, intermediate 15-1 (547 mg, 2.13 mmol, 1 eq) was added and stirred at room temperature for 5 min. A saturated ammonium chloride solution was added dropwise, and stirred at room temperature. Thin layer chromatography was performed until the reaction was complete, and the reaction was filtered off and the tetrahydrofuran was removed by vacuum rotary evaporation. The reaction was extracted with saturated brine and ethyl acetate, and the organic phase was dried over anhydrous magnesium sulfate. The filtrate was concentrated by suction and the compound was obtained by column chromatography. A white solid was obtained with a yield of 85%. 1 H NMR (400MHz, Acetone) δ7.80(d,J=2.0Hz,1H),7.77(d,J=8.9Hz,1H),7.45(s,1H),7.27(dd,J=8.8,2.0Hz,1H),6.02(d,J=1.5Hz,1H) ,5.42(s,1H),5.10(t,J=5.5Hz,1H),4.91(d,J=4.8Hz,1H),4.28(t,1H),4.20(dd,J=9.4,3.9Hz,1H),3.62-3.57(m,1H),3.22(s,3H). 13 C NMR (101MHz, Acetone) δ171.02,136.53,134.93,130.92,129.39,126.40,125.69,124.10,123.41,121.22,115.56,69.67,68.90,44.76,41.29.

[0051] Example 14 Synthesis of Compound 14 4-((5-Chloro-1-(propylsulfonyl)-1H-indol-3-yl)(hydroxy)methyl)-3-methylenedihydrofuran-2(3H)-one

[0052] The synthesis process was the same as in Example 13, except that R1A was replaced with propyl acyl to obtain a white solid with a yield of 65%. 1 H NMR (400MHz, Acetone) δ7.93(d,J=1.9Hz,1H),7.91(d,J=8.9Hz,1H),7.59(s,1H),7.39(dd,J=8.9,2.0Hz,1H),6.16(d,J=1.3Hz,1H),5.57(s,1H),5.24 (t,1H),5.07(d,J=4.7Hz,1H),4.42(t,1H),4.34(dd,J=9.4,3.9Hz,1H),3.7 8-3.73(m,1H),3.53-3.49(m,2H),1.70-1.61(m,2H),0.96(t,J=7.4Hz,3H). 13 C NMR (101MHz, Acetone) δ170.97,136.61,135.07,130.77,129.27,126.83,125. 61,124.02,122.88,121.21,115.59,69.68,68.8,56.27,44.79,17.88,12.62.

[0053] Example 15 Synthesis of Compound 15 4-((5-Chloro-1-((2-fluorophenyl)sulfonyl)-1H-indol-3-yl)(hydroxy)methyl)-3-methylenedihydrofuran-2(3H)-one

[0054] The synthesis process was the same as in Example 13, except that the R1 group was replaced with 2-fluorobenzenesulfonyl to obtain a white solid with a yield of 68%. 1 HNMR(400MHz,DMSO-d6)δ8.14(td,J=7.7,1.6Hz,1H),7.92(d,J=2.0Hz,1H),7.86-7 .82(m,1H),7.82-7.78(m,1H),7.65(s,1H),7.49(td,J=8.8,4.4Hz,2H),7.39(dd,J= 8.9,2.1Hz,1H),5.97(d,J=1.5Hz,1H),5.93(d,J=3.9Hz,1H),5.09(d,J=0.7Hz,1H) ,5.06(s,1H),4.36(t,J=8.7Hz,1H),4.22(dd,J=9.2,3.7Hz,1H),3.55-3.47(m,1H). 13C NMR(101MHz,DMSO-d6)δ170.42,158.16,138.10,135.06,132.96,130.31,129.74,128.35,12 5.72,125.49,124.97,124.55,123.37,123.19,120.55,117.99,114.61,68.36,67.71,43.01.

[0055] Example 16 Synthesis of Compound 16 4-((5-Chloro-1-((4-fluorophenyl)sulfonyl)-1H-indol-3-yl)(hydroxy)methyl)-3-methylenedihydrofuran-2(3H)-one

[0056] The synthesis process was the same as in Example 13, except that the R1 group was replaced with 4-fluorobenzenesulfonyl to obtain a white solid with a yield of 71%. 1 HNMR (400MHz, Acetone-d6) δ8.16–8.09(m,2H),8.05(d,J=8.9Hz,1H),7.85(d,J=2.0Hz,1H),7.74(s,1H),7.50–7.3 6(m,3H),6.05(s,1H),5.24-5.20(m,2H),5.05(d,J=4.8Hz,1H),4.44–4.35(m,1H),4.34–4.30(m,1H),3.66(s,1H). 13 C NMR(100MHz,Acetone)δ170.93,168.16,165.63,136.30,134.87,134.57,131.33,130.95,12 9.85,126.54,126.04,124.94,123.81,121.30,118.01,117.78,116.00,69.64,68.96,44.46.

[0057] Example 17 Synthesis of Compound 17 4-((5-Chloro-1-((2-chlorophenyl)sulfonyl)-1H-indol-3-yl)(hydroxy)methyl)-3-methylenedihydrofuran-2(3H)-one

[0058] The synthesis process was the same as in Example 13, except that the R1 group was replaced with 2-chlorobenzenesulfonyl to obtain a white solid with a yield of 71%. 1HNMR(400MHz,MeOD)δ8.25(dd,J=8.2,1.6Hz,1H),7.79(d,J=2.0Hz,1H),7.76(s ,1H),7.70(d,J=8.9Hz,1H),7.68-7.62(m,1H),7.60-7.55(m,2H),7.26(dd,J=8 .9,2.0Hz,1H),6.17(d,J=2.1Hz,1H),5.48(d,J=1.5Hz,1H),5.05(d,J=6.2Hz,1 H),4.38(dd,J=9.2,8.3Hz,1H),4.30(dd,J=9.4,3.9Hz,1H),3.65-3.60(m,1H). 13 C NMR(101MHz,DMSO-d6)δ170.39,136.50,135.23,134.57,132.92,132.65,131.52,131.28,12 9.68,128.48,128.12,126.47,124.79,123.47,122.24,120.62,114.34,68.19,67.69,43.16.

[0059] Example 18 Synthesis of Compound 18 4-((5-Chloro-1-((3-chlorophenyl)sulfonyl)-1H-indol-3-yl)(hydroxy)methyl)-3-methylenedihydrofuran-2(3H)-one

[0060] The synthesis process was the same as in Example 13, except that the R1 group was replaced with 3-chlorobenzenesulfonyl to obtain a white solid with a yield of 82%. 1 HNMR(400MHz,DMSO-d6)δ8.06(t,J=1.8Hz,1H),8.01(d,J=8.9Hz,1H),7.94(d,J=8.0Hz,1 H),7.89(d,J=2.0Hz,1H),7.82(dd,J=8.1,1.2Hz,1H),7.74(s,1H),7.64(t,J=8.0Hz,1H), 7.43(dd,J=8.9,2.0Hz,1H),5.95(d,J=1.4Hz,1H),5.90(d,J=5.0Hz,1H),5.02(d,J=5.2H z,1H),5.00(s,1H),4.33(t,J=8.6Hz,1H),4.20(dd,J=9.2,3.7Hz,1H),3.53-3.44(m,1H). 13C NMR(101MHz,DMSO-d6)δ170.37,138.21,135.09,134.90,134.50,133.19,131.95,130.03,12 8.49,126.19,125.51,125.43,125.23,124.45,123.09,120.52,115.00,68.31,67.69,42.91.

[0061] Example 19 Synthesis of Compound 19 4-((5-Chloro-1-((4-chlorophenyl)sulfonyl)-1H-indol-3-yl)(hydroxy)methyl)-3-methylenedihydrofuran-2(3H)-one

[0062] The synthesis process was the same as in Example 13, except that the R1 group was replaced with 4-chlorobenzenesulfonyl to obtain a white solid with a yield of 80%. 1 HNMR(400MHz,DMSO-d6)δ7.98(d,J=8.5Hz,3H),7.89(s,1H),7.78-7.63(m,3H),7.42(d,J=8.8Hz,1H), 5.96(s,1H),5.89(d,J=4.8Hz,1H),5.02(s,2H),4.35(t,J=8.6Hz,1H),4.23-4.20(m,1H),3.49(s,1H). 13 C NMR(100MHz,DMSO-d6)δ170.31,139.99,135.30,135.03,133.16,130.11,129.97,12 8.58,128.40,125.41,125.16,124.30,123.23,120.49,114.91,68.29,67.66,42.95.

[0063] Example 20 Synthesis of Compound 20 4-((1-((4-bromophenyl)sulfonyl)-5-chloro-1H-indol-3-yl)(hydroxy)methyl)-3-methylenedihydrofuran-2(3H)-one

[0064] The synthesis process was the same as in Example 13, except that the R1 group was replaced with 4-bromobenzenesulfonyl to obtain a white solid with a yield of 80%. 1HNMR (400MHz, DMSO-d6) δ7.97(d,J=8.9Hz,1H),7.89(dd,J=5.5,3.3Hz,3H),7.84(d,J=8.8Hz,2H),7.67(s,1H),7.42(dd,J=8.9,2.0Hz,1H) ,5.95(d,J=1.4Hz,1H),5.88(d,J=5.0Hz,1H),5.02(d,J=7.4Hz,2H),4.34(t,J=8.7Hz,1H),4.21(dd,J=9.2,3.6Hz,1H),3.52-3.45(m,1H). 13 C NMR(101MHz,DMSO-d6)δ170.37,135.74,135.05,133.18,133.06,130.02,129.16,12 8.55,128.42,125.41,125.17,124.33,123.24,120.50,114.92,68.30,67.67,42.96.

[0065] Example 21 Synthesis of Compound 21 4-((5-Chloro-1-((2-nitrophenyl)sulfonyl)-1H-indol-3-yl)(hydroxy)methyl)-3-methylenedihydrofuran-2(3H)-one

[0066] The synthesis process was the same as in Example 13, except that the R1 group was replaced with 2-nitrobenzenesulfonyl to obtain a white solid with a yield of 26%. 1 H NMR (400MHz, CDCl3) δ8.02 (dd, J=6.3, 3.2Hz, 1H), 7.83-7.73 (m, 4H), 7.72 (d ,J=1.9Hz,1H),7.60(s,1H),7.32(dd,J=8.9,2.0Hz,1H),6.40(d,J=2.1Hz,1 H),5.91(d,J=1.8Hz,1H),4.96(dd,J=7.9,3.7Hz,1H),4.30(dd,J=9.7,8.2H z,1H),4.19(dd,J=9.8,4.0Hz,1H),3.65-3.59(m,1H),2.62(d,J=3.9Hz,1H). 13C NMR (101MHz, CDCl3) δ170.66,147.93,135.70,134.58,133.54,132.87,131.06,130.56,130 .15,129.75,126.40,126.22,125.95,125.33,121.37,120.62,114.75,69.19,67.81,44.14.

[0067] Example 22 Synthesis of Compound 22 4-((5-chloro-1-o-methylbenzenesulfonyl)-1H-indol-3-yl)(hydroxy)methyl)-3-methylenedihydrofuran-2(3H)-one

[0068] The synthesis process was the same as in Example 13, except that the R1 group was replaced with 2-methylbenzenesulfonyl to obtain a white solid with a yield of 41%. 1 H NMR (400MHz, CDCl3) δ7.96-7.85(m,1H),7.76-7.61(m,3H),7.55-7.47(m,1H),7.39-7 .32(m,1H),7.29(dd,J=8.6,4.8Hz,2H),7.24(d,J=2.0Hz,1H),6.32(dd,J=28.1,2.0H z,1H),5.66(dd,J=89.6,1.7Hz,1H),4.94(dd,J=13.9,7.3Hz,1H),4.31-4.20(m,1H), 4.18-4.08(m,1H),3.62-3.45(m,1H),2.45(d,J=52.5Hz,3H),2.20(d,J=131.6Hz,1H). 13 C NMR(101MHz,DMSO-d6)δ170.41,137.41,135.30,134.96,133.58-132.89,130.32,129.63,128.99,128.06,127.1 9,126.69,125.92,125.53,124.88,123.61,122.36,120.57,120.36,114.92,114.40,68.19,67.71,43.19,19.69.

[0069] Example 23 Synthesis of Compound 23 4-((5-Chloro-1-toluenesulfonyl)-1H-indol-3-yl)(hydroxy)methyl)-3-methylenedihydrofuran-2(3H)-one

[0070] The synthesis process was the same as in Example 13, except that the R1 group was replaced with 4-methylbenzenesulfonyl to obtain a white solid with a yield of 69%. 1 H NMR (400MHz, DMSO-d6) δ7.97(d,J=8.9Hz,1H),7.87(d,J=2.0Hz,1H),7.85(s,1H),7.83(s,1H),7.67(s,1H),7.42(s,1H),7.41-7.39(m,2H),5.9 6(d,J=1.6Hz,1H),5.88(d,J=5.0Hz,1H),5.06-4.98(m,2H),4.34(t,J= 8.7Hz,1H),4.21(dd,J=9.2,3.7Hz,1H),3.49-3.47(m,1H),2.33(s,3H). 13 C NMR(101MHz,DMSO-d6)δ170.42,145.79,135.04,133.75,133.22,130.32,129.93,128.1 4,126.68,125.53,124.94,123.76,123.28,120.36,114.92,68.35,67.72,42.98,21.02.

[0071] Example 24 Synthesis of Compound 24 4-((5-Chloro-1-((4-methoxyphenyl)sulfonyl)-1H-indol-3-yl)(hydroxy)methyl)-3-methylenedihydrofuran-2(3H)-one

[0072] The synthesis process was the same as in Example 13, except that the R1 group was replaced with 4-methoxybenzenesulfonyl to obtain a white solid with a yield of 42%. 1 H NMR (400MHz, DMSO-d6) δ7.96(d,J=8.9Hz,1H),7.90(d,J=2.0Hz,1H),7.88(d,J=2.0Hz ,1H),7.87(d,J=2.1Hz,1H),7.65(s,1H),7.40(dd,J=8.9,2.1Hz,1H),7.14-7.11(m,1H ),7.11-7.09(m,1H),5.97(d,J=1.6Hz,1H),5.86(d,J=5.1Hz,1H),5.01(t,J=4.9Hz,2 H),4.34(t,J=8.7Hz,1H),4.20(dd,J=9.2,3.7Hz,1H),3.80(s,3H),3.51-3.43(m,1H).13 C NMR(101MHz,DMSO-d6)δ170.41,163.94,135.05,133.19,129.90,129.12,128.03,128.0 0, 125.53, 124.86, 123.55, 123.30, 120.31, 115.07, 114.89, 68.34, 67.72, 55.92, 43.00.

[0073] Example 25 Synthesis of Compound 25 4-((5-Chloro-1-(3-(trifluoromethyl)phenyl)sulfonyl)-1H-indol-3-yl)(hydroxy)methyl)-3-methylenedihydrofuran-2(3H)-one

[0074] The synthesis process was the same as in Example 13, except that the R1 group was replaced with 3-trifluoromethylbenzenesulfonyl to obtain a white solid with a yield of 53%. 1 H NMR (400MHz, DMSO-d6) δ8.29(d,J=6.3Hz,2H),8.14(d,J=7.9Hz,1H),8.05(d,J=8.9Hz,1H),7.91-7.85(m,2H),7.80(s,1H),7.44(dd,J=8.9,2. 1Hz,1H),5.91(dd,J=5.6,3.3Hz,2H),5.04(s,1H),5.00(t,J=5.3Hz,1H),4.30(t,J=8.7Hz,1H),4.17(dd,J=9.2,3.8Hz,1H),3.52-3.43(m,1H). 13 C NMR(101MHz,DMSO-d6)δ170.32,137.69,135.16,133.17,131.89-131.34,130.75,130.4,130.11 ,128.56,125.46,125.26,124.62,123.30,123.09,122.93,120.60,114.95,68.18,67.63,42.89.

[0075] Example 26 Synthesis of Compound 26 4-((5-Chloro-1-((4-(trifluoromethyl)phenyl)sulfonyl)-1H-indol-3-yl)(hydroxy)methyl)-3-methylenedihydrofuran-2(3H)-one

[0076] The synthesis process was the same as in Example 13, except that the R1 group was replaced with 4-trifluoromethylbenzenesulfonyl to obtain a white solid with a yield of 69%. 1 H NMR (400MHz, DMSO-d6) δ8.19(d,J=8.2Hz,2H),8.02(d,J=8.2Hz,3H),7.90(s,1H),7.71(s,1H),7.44(d,J=8.8Hz,1H),5.9 3(s,1H),5.90(d,J=4.6Hz,1H),5.04(s,1H),4.97(s,1H),4.34(t,J=8.6Hz,1H),4.22(dd,J=9.0,2.8Hz,1H),3.48(s,1H). 1 13 C NMR (100MHz, DMSO) δ170.33,140.22,135.02,134.29,133.97,133.19,130.05,128.58,127. 73,127.20,125.34,124.66,124.35,123.12,121.65,120.58,114.93,68.28,67.64,42.91.

[0077] Example 27 Synthesis of Compound 27 4-((5-Chloro-1-((4-(trifluoromethoxy)phenyl)sulfonyl)-1H-indol-3-yl)(hydroxy)methyl)-3-methylenedihydrofuran-2(3H)-one

[0078] The synthesis process was the same as in Example 13, except that the R1 group was replaced with 4-trifluoromethoxybenzenesulfonyl to obtain a white solid with a yield of 69%. 1 H NMR (400MHz, DMSO-d6) δ8.19(d,J=8.2Hz,2H),8.02(d,J=8.2Hz,3H),7.90(s,1H),7.71(s,1H),7.44(d,J=8.8Hz,1H),5.9 3(s,1H),5.90(d,J=4.6Hz,1H),5.04(s,1H),4.97(s,1H),4.34(t,J=8.6Hz,1H),4.22(dd,J=9.0,2.8Hz,1H),3.48(s,1H). 13C NMR(100MHz,DMSO-d6)δ170.33,140.22,135.02,133.92,133.19,130.05,128.58,127.73 ,127.22,127.18,125.34,124.66,123.12,121.65,120.58,114.93,68.28,67.64,42.78.

[0079] Example 28 Synthesis of Compound 28 4-((1-((4-(bromomethyl)phenyl)sulfonyl)-5-chloro-1H-indol-3-yl)(hydroxy)methyl)-3-methylenedihydrofuran-2-yl (3H)-Keto

[0080] The synthesis process was the same as in Example 13, except that the R1 group was replaced with 4-bromomethylbenzenesulfonyl to obtain a white solid with a yield of 75%. 1 H NMR (400MHz, DMSO) δ7.96(d,J=8.8Hz,1H),7.87(s,1H),7.84(d,J=8.1Hz,2H),7.67(s,1H),7.42-7.39(m,3H ),5.96(s,1H),5.88(s,1H),5.01(s,2H),4.34(t,J=8.6Hz,1H),4.22-4.18(m,1H),3.48(s,1H),2.33(s,2H). 13 C NMR (100MHz, CDCl3) δ170.07,153.88,135.67,134.28,133.84,130.04,129.45,129.12 ,126.11,125.66,125.08,122.11,121.16,120.19,114.99,69.54,67.69,47.31,44.08.

[0081] Example 29 Synthesis of Compound 29 4-((5-Chloro-1-((3,4-dichlorophenyl)sulfonyl)-1H-indol-3-yl)(hydroxy)methyl)-3-methylenedihydrofuran-2(3H)-one

[0082] The synthesis process was the same as in Example 13, except that the R1 group was replaced with 3,4-dichlorobenzenesulfonyl to obtain a white solid with a yield of 76%. 1H NMR (400MHz, DMSO-d6) δ8.31(d,J=2.0Hz,1H),8.02(d,J=8.9Hz,1H),7.93(dd,J =8.6,2.1Hz,1H),7.91-7.88(m,2H),7.75(s,1H),7.43(dd,J=8.9,2.1Hz,1H),5 .97(d,J=1.4Hz,1H),5.90(d,J=5.0Hz,1H),5.07(d,J=0.7Hz,1H),5.01(t,J=5. 2Hz, 1H), 4.34 (t, J = 8.7Hz, 1H), 4.21 (dd, J = 9.2, 3.7Hz, 1H), 3.54-3.45 (m, 1H). 13 C NMR(100MHz,DMSO-d6)δ170.33,138.29,136.48,135.09,133.17,132.97,132.25,130.11,12 8.58,128.48,126.69,125.49,125.27,124.62,123.17,120.55,115.02,68.25,67.65,42.95.

[0083] Example 30 Synthesis of Compound 30 4-((5-chloro-1-((4-chloro-3-nitrophenyl)sulfonyl)-1H-indol-3-yl)(hydroxy)methyl)-3-methylenedihydrofuran-2-yl(3H) -ketone

[0084] The synthesis process was the same as in Example 13, except that the R1 group was replaced with 3-nitro-4-chlorobenzenesulfonyl to obtain a white solid with a yield of 17%. 1 H NMR(400MHz,DMSO-d6)δ8.76(d,J=2.2Hz,1H),8.25(dd,J=8.6,2.3Hz,1H),8.03 (dd,J=8.7,4.5Hz,2H),7.90(d,J=2.0Hz,1H),7.76(s,1H),7.44(dd,J=8.9,2.0 Hz,1H),5.98(d,J=1.4Hz,1H),5.90(d,J=5.0Hz,1H),5.13(s,1H),5.02(t,J=5. 2Hz, 1H), 4.34 (t, J = 8.7Hz, 1H), 4.22 (dd, J = 9.3, 3.6Hz, 1H), 3.53-3.44 (m, 1H). 13C NMR(100MHz,DMSO-d6)δ170.32,147.87,136.30,135.01,133.73,133.07,131.77,131.11,13 0.18,128.71,125.37,125.33,124.86,124.06,123.38,120.62,115.02,68.19,67.62,42.98.

[0085] Example 31 Synthesis of Compound 31 4-((5-Chloro-1-((2,3-dihydrobenzo[b][1,4]dioxin-6-yl)sulfonyl)-1H-indol-3-yl)(hydroxy)methyl)-3-methylenedihydrofuran-2(3H)-one

[0086] The synthesis process was the same as in Example 13, except that the R1 group was replaced with 2,3-dihydro-1,4-benzodioxy-6-sulfonyl to obtain a white solid with a yield of 96.85%. 1 H NMR(400MHz,DMSO-d6)δ7.96(d,J=8.9Hz,1H),7.87(d,J=2.0Hz,1H),7.67(s,1H ),7.42(m,J=8.5,2.2Hz,3H),7.05(d,J=8.4Hz,1H),5.97(d,J=1.5Hz,1H),5.87( d,J=5.1Hz,1H),5.05(d,J=0.7Hz,1H),5.01(t,J=5.1Hz,1H),4.34(t,J=8.7Hz,1 H),4.28(dd,J=10.8,4.7Hz,4H),4.20(dd,J=9.2,3.7Hz,1H),3.51-3.46(m,1H). 13 C NMR(101MHz,DMSO-d6)δ170.54,148.97,143.71,135.11,133.29,129.97,128.65,128.19,125.71,1 25.03,123.62,123.34,120.64,120.40,118.24,115.69,115.03,68.44,67.73,64.57,64.07,43.04.

[0087] Example 32 Synthesis of Compound 32 4-((5-chloro-1-((5-chloro-2,4-difluorophenyl)sulfonyl)-1H-indol-3-yl)(hydroxy)methyl)-3-methylenedihydrofuran-2(3H)-one

[0088] The synthesis process was the same as in Example 13, except that the R1 group was replaced with 5-chloro-2,4-difluorobenzenesulfonyl to obtain a white solid with a yield of 65%. 1 H NMR(400MHz, DMSO-d6)δ8.48(t,J=7.3Hz,1H),8.02-7.85(overlap,3H),7.68(s,1H),7.42(d,J=8.9Hz,1H) ,6.01(s,1H),5.93(s,1H),5.16(s,1H),5.06(s,1H),4.37(t,J=8.6Hz,1H),4.25-4.22(m,1H),3.51(s,1H). 13 C NMR(100MHz,DMSO-d6)δ170.36,162.92,156.26,134.99,132.90,131.68,125.35,125.14,123.88 ,123.27,122.30,122.11,120.56,117.27,117.04,114.83,108.80,118.26,68.28,67.66,43.05.

[0089] Example 33 Synthesis of Compound 33 4-((5-Chloro-1-((4-fluoro-3-(trifluoromethyl)phenyl)sulfonyl)-1H-indol-3-yl)(hydroxy)methyl)-3-methylenedihydrofuran-2(3H)-one

[0090] The synthesis process was the same as in Example 13, except that the R1 group was replaced with 4-fluoro-3-trifluoromethylbenzenesulfonyl to obtain a white solid with a yield of 70%. 1 H NMR (400MHz, Acetone-d6) δ8.47–8.41(m,1H),8.39-8.37(m,1H),8.10(d,J=8.9Hz,1H),7.87(d,J=2.0Hz,1H),7.83(s,1H),7.72(t,J=9.5Hz,1H),7. 43(dd,J=8.9,2.0Hz,1H),6.06(s,1H),5.28(s,1H),5.23(t,J=5.6Hz,1H) ,5.09(d,J=4.8Hz,1H),4.40-4.36(m,1H),4.34-4.31(m,1H),3.66(s,1H). 13C NMR (100MHz, Acetone-d6) δ169.91,162.83,135.43,134.35,134.03,130.50,129.25,126.95,126. 91,126.88,125.46,124.68,123.07,122.92,120.60,120.36,119.44,115.10,68.69,68.06,43.50.

[0091] Example 34 Synthesis of Compound 34 4-((5-Chloro-1-((3-nitro-4-(propylamino)phenyl)sulfonyl)-1H-indol-3-yl)(hydroxy)methyl)-3-methylenedihydrofuran-2(3H)-one

[0092] The synthesis process is the same as in Example 13, the R1 group is replaced with 3-nitro-4-chlorobenzenesulfonyl to obtain intermediate 36-1, and then the intermediate 36-2 is obtained by substitution reaction with n-propylamine under alkaline conditions. The final synthesis steps are the same as in Example 15 to obtain a yellow solid with a yield of 7%. 1 H NMR (400MHz, DMSO-d6) δ8.60(t,J=5.7Hz,1H),8.46(d,J=2.3Hz,1H),7.89(d,J=8.9Hz,1H),7.81(d,J=1. 9Hz,1H),7.76(dd,J=9.3,2.2Hz,1H),7.63(s,1H),7.36(dd,J=8.8,1.9Hz,1H),7.11(d,J=9.4Hz,1H),5. 89(d,J=1.1Hz,1H),5.79(d,J=5.1Hz,1H),5.04(s,1H),4.95(t,J=5.2Hz,1H),4.26(t,J=8.7Hz,1H),4.1 4(dd,J=9.2,3.6Hz,1H),3.45-3.38(m,1H),3.29-3.24(m,2H),1.54-1.44(m,2H),0.81(t,J=7.4Hz,3H). 13C NMR (100MHz, DMSO-d6) δ170.41,147.75,135.14,133.18,132.26,130.06,129.94,128.21,126.84,125. 54,125.09,123.88,123.28,120.96,120.46,116.29,114.87,68.31,67.72,44.13,43.05,21.30,11.07.

[0093] Example 35 Synthesis of Compound 35 4-((5-Chloro-1-((4-(isobutylamino)-3-nitrophenyl)sulfonyl)-1H-indol-3-yl)(hydroxy)methyl)-3-methylenedihydrofuran-2(3H)-one

[0094] The synthesis process is the same as Example 34, except that the R1 group is replaced with 3-nitro-4-(isobutylamino)benzenesulfonyl to obtain a yellow solid with a yield of 11%. 1 H NMR (400MHz, CDCl3) δ8.74(d,J=2.3Hz,1H),8.53(t,J=5.0Hz,1H),7.92(d,J=8.9Hz,1H),7.73(dd,J=9.2 ,2.1Hz,1H),7.65(d,J=1.8Hz,1H),7.56(s,1H),7.35(dd,J=8.9,1.9Hz,1H),6.86(d,J=9.3Hz,1H),6.37 (d,J=1.9Hz,1H),5.73(d,J=1.5Hz,1H),4.95(d,J=7.4Hz,1H),4.26(dd,J=9.5,8.1Hz,1H),4.16(dd,J=9 .7,3.8Hz,1H),3.58-3.53(m,1H),3.12(dd,J=6.6,5.5Hz,2H),2.04-1.93(m,1H),1.03(d,J=6.7Hz,6H). 13 C NMR (100MHz, CDCl3) δ170.55,148.36,134.44,133.71,132.76,130.71,129.83,129.65,127.82,125.9 8,125.88,125.36,122.79,121.85,120.22,115.25,115.02,69.47,67.91,51.02,44.10,27.98,20.36.

[0095] Example 36 Synthesis of Compound 36 4-((5-chloro-1-((3-nitro-4-(piperazin-1-yl)phenyl)sulfonyl)-1H-indol-3-yl)(hydroxy)methyl)-3-methylenedihydrofuran-2(3H)-one

[0096] The synthesis process is the same as Example 34, except that the R1 group is replaced with 3-nitro-4-piperazinylbenzenesulfonyl to obtain a yellow solid. 1 HNMR (400MHz, Acetone) δ8.44 (d, J = 2.3Hz, 1H), 8.10-8.07 (m, 1H), 8.07-8.05 (m, 1H ),7.86(d,J=1.9Hz,1H),7.77(s,1H),7.53(d,J=9.0Hz,1H),7.40(dd,J=8.9,2.0Hz, 1H),6.08(d,J=1.3Hz,1H),5.32(s,1H),5.22(d,J=5.8Hz,1H),4.41-4.35(m,1H),4 .32(dd,J=9.4,3.9Hz,1H),3.69-3.65(m,1H),3.64-3.60(m,4H),3.54-3.47(m,4H). 13 C NMR(100MHz,Acetone)δ170.89,149.53,140.44,136.32,134.70,132.36,131.40,129.94,128.92,1 27.08,126.56,126.13,125.01,123.98,122.92,121.42,116.03,69.51,68.86,48.29,44.56,43.72.

[0097] Example 37 Synthesis of Compound 37 4-((5-chloro-1-((4-(4-methylpiperazin-1-yl)-3-nitrophenyl)sulfonyl)-1H-indol-3-yl)(hydroxy)methyl)-3-methylenedihydrofuran-2(3H)-one

[0098] The synthesis process is the same as Example 34, except that the R1 group is replaced with 4-(4-methylpiperazine)-3-nitrobenzenesulfonyl to obtain a yellow solid with a yield of 11%. 1H NMR (400MHz, DMSO-d6) δ8.45-8.42(m,1H),8.08-7.98(m,2H),7.89(d,J=1.7Hz,1H),7.75(m,1H),7.53-7.41(m,2H),6.00(s,1H),5.1 4(s,1H),5.02(d,J=5.5Hz,1H),4.34(t,J=8.6Hz,1H),4.22(dd,J=9.2,3.5Hz,1H),3.83-3.68(m,4H),3.58-3.46(m,5H),2.82(s,3H). 13 C NMR (100MHz, DMSO-d6) δ170.40,147.75,138.35,135.06,133.11,131.26,130.13,128.37,126.53,126. 12,125.54,125.16,124.11,122.24,120.52,118.36,114.98,68.26,67.65,51.82,47.02,43.05,42.07.

[0099] Example 38 Synthesis of Compound 38 tert-Butyl (2-((4-((5-chloro-3-(hydroxy(4-methylene-5-oxotetrahydrofuran-3-yl)methyl)-1H-indol-1-yl)sulfonyl)-2-nitrophenyl)amino)ethyl)carbamate

[0100] The synthesis process was the same as in Example 34, except that the R1 group was replaced with tert-butyl-(2-((4-sulfonyl)-2-nitrophenyl)amine)ethyl)carbamate to obtain a yellow-green solid with a yield of 71%. 1 H NMR (400MHz, Acetone) δ8.69(d,J=2.4Hz,1H),8.06(d,J=8.8Hz,1H),7.93(d,J=9.1Hz,1H),7 .87(d,J=2.1Hz,1H),7.79(s,1H),7.43(dd,J=8.8,2.1Hz,1H),7.32(d,J=9.4Hz,1H),6.31(s, 1H),6.10(s,1H),5.33(s,1H),5.23(t,J=5.5Hz,1H),5.03(d,J=4.9Hz,1H),4.51–4.28(m,2H) ,3.64(dd,J=15.9,10.1Hz,3H),3.39(dd,J=12.1,6.0Hz,2H),2.80(s,1H),1.58–1.04(m,9H).

[0101] Example 39 Synthesis of Compound 39 tert-Butyl (4-((4-((5-chloro-3-(hydroxy(4-methylene-5-oxotetrahydrofuran-3-yl)methyl)-1H-indol-1-yl)sulfonyl)-2-nitrophenyl)amino)butyl)carbamate

[0102] The synthesis process was the same as in Example 34, except that the R1 group was replaced with tert-butyl-(2-((4-sulfonyl)-2-nitrophenyl)amino)butyl)carbamate to obtain a yellow-green solid with a yield of 76%. 1 H NMR (400MHz, Acetone-d6) δ8.68(d,J=2.3Hz,1H),8.64(s,1H),8.04(d,J=8.9Hz,1H),7.91(dd,J=9. 3,2.3Hz,1H),7.85(d,J=2.0Hz,1H),7.76(s,1H),7.41(dd,J=8.9,2.0Hz,1H),7.25(d,J=9.3Hz,1H) ,6.08(s,1H),5.98(s,1H),5.31(s,1H),5.22(t,J=5.4Hz,1H),5.04(d,J=4.9Hz,1H),4.41-4.35(m, 1H),4.32-4.29(m,1H),3.66(s,1H),3.53(dd,J=13.2,6.9Hz,2H),3.09(dd,J=5.4,2.3Hz,2H),1.79 -1.67(overlap,2H),1.59(dd,J=14.7,7.1Hz,2H),1.37(s,9H). 13 C NMR (100MHz, Acetone-d6) δ170.93,149.15,136.35,134.75,133.44,131.52,131.35,129.73,128.07,126.61,126.01,1 24.66,123.95,123.14,121.32,116.80,115.92,78.49,69.66,68.90,44.37,43.61,40.61,28.64,28.13,26.57,14.31.

[0103] Example 40 Synthesis of Compound 40 4-(5-(4-(4-((5-chloro-3-(hydroxy(4-methylene-5-oxotetrahydrofuran-3-yl)methyl)-1H-indol-1-yl)sulfonyl)benzyl)piperazin-1-yl)-5-oxopentyl)tetrahydro-1H-thieno[3,4-d]imidazol-2(3H)-one

[0104] The synthesis process is the same as in Example 34, except that intermediate 28-1 is used to replace compound 34-1 to obtain a white solid. Yield 65%. 1 H NMR (400MHz, Acetone-d6) δ8.07(d,J=8.9Hz,1H),7.97(d,J=8.4Hz,2H),7.87(s,1H),7.84(d ,J=2.0Hz,1H),7.73(s,1H),7.63(d,J=8.4Hz,2H),7.40(dd,J=8.9,2.0Hz,1H),7.33(s,1H),6 .98(s,1H),6.01(s,1H),5.63(s,1H),5.25-5.21(m,1H),5.14-5.12(m,1H),5.11-5.07(m,1H) ,4.40-4.32(m,1H),4.31-4.28(m,1H),3.67-3.62(m,3H),3.62–3.55(m,4H),2.86(s,3H),2.6 -2.43(m,4H),2.17-2.00(m,3H),1.34-1.18(m,1H). 13 C NMR (100MHz, Acetone-d6) δ194.57,187.26,170.93,151.34,147.02,137.68,137.16,136.23,134.96,131.14,130.70,129.92,129.71,1 27.88,126.63,125.94,124.67,123.75,123.17,121.22,119.04,116 .07,69.65,69.09,62.27,54.91,53.35,46.95,44.51,28.50,27.75. Examples 41-44 Synthesis of Compounds 41-44 Synthesis of compound 41 1-(5-Chloro-1-((4-chlorophenyl)sulfonyl)-1H-indol-3-yl)but-3-yn-1-ol

[0105] Bromomethylbutenolide (265.50 mg, 1.50 mmol) was dissolved in 3 ml of tetrahydrofuran, and zinc powder (326.95 mg, 5.01 mmol) was added and stirred at room temperature for about 10 min. Compound 19-1 (354.20 mg, 1.02 mmol) was added and stirred at room temperature for 5 min. About 1 ml of saturated ammonium chloride solution was slowly added dropwise and stirred at room temperature. The reaction was detected by TLC until complete, and the mixture was filtered and THF was removed by vacuum rotary evaporation. The mixture was extracted with saturated brine and ethyl acetate. The organic phase was dried over anhydrous magnesium sulfate, filtered and the filtrate was concentrated. Compound 41 was obtained by column chromatography as a white solid with a yield of 66%. 1 H NMR (400MHz, CDCl3) δ7.88(d,J=8.8Hz,1H),7.78(d,J=8.6Hz,2H),7.64(d,J=1.9Hz,1H),7.58(s,1H),7. 41(d,J=8.6Hz,2H),7.32-7.25(m,1H),5.06(s,1H),2.80-2.75(m,2H),2.47(s,1H),2.11(t,J=1Hz,1H). 13 C NMR (101MHz, CDCl3) δ140.94,136.11,133.66,130.03,129.76,129.61,128 .18,125.51,124.26,123.80,120.30,114.72,79.69,72.13,65.84,27.63. 13 C NMR (100MHz, CDCl3) δ140.94,136.11,133.66,130.03,129.76,129.61,128 .18,125.51,124.26,123.80,120.30,114.72,79.69,72.13,65.84,27.63. Synthesis of compound 42 N-((5-Chloro-1-((4-chlorophenyl)sulfonyl)-1H-indol-3-yl)methyl)cyclopropylamine

[0106] Intermediate 19-1 (177.10 mg, 0.50 mmol) was dissolved in 10 ml of dichloromethane, and cyclopropylamine (85.64 mg, 1.50 mmol) and magnesium sulfate (37.83 mg, 1.50 mmol) were added in sequence, stirred at room temperature, and the reaction was detected by TLC. After filtration, the organic phase was concentrated, 3 ml of anhydrous methanol was added to dissolve, NaBH4 (1.00 mmol) was added under ice bath conditions and stirred to react, and the reaction was monitored by TLC to be complete. The pH was adjusted to neutral with 10% HCl, and the anhydrous methanol was removed by vacuum rotary evaporation, and extracted with saturated brine and dichloromethane. The organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and column chromatography was performed to obtain compound 42 as a white solid with a yield of 50.03%. 1 HNMR (400MHz, CDCl3) δ7.90(d,J=8.8Hz,1H),7.79(d,J=8.6Hz,2H),7.56(d,J=2.0Hz,1H),7.47(s,1H),7.42(d, J=8.6Hz,2H),7.30(dd,J=8.8,2.0Hz,1H),3.92(s,2H),2.17(s,1H),1.38(d,J=15.1Hz,1H),0.56–0.35(m,4H). 13 C NMR (101MHz, CDCl3) δ188.73,140.81,136.37,133.80,131.79,129.68,129.44 ,128.12,125.35,124.77,122.11,119.70,114.71,44.07,30.45,29.90,6.26. Synthesis of compound 43 1-(5-Chloro-1-((4-chlorophenyl)sulfonyl)-1H-indol-3-yl)-N-(cyclopropylmethyl)methanamine

[0107] Intermediate 19-1 (177.10 mg, 0.50 mmol) was dissolved in 10 ml of dichloromethane, and cyclomethamine (106.68 mg, 1.50 mmol) and magnesium sulfate (37.83 mg, 1.50 mmol) were added in sequence, stirred at room temperature, and the reaction was detected by TLC. After suction filtration, the organic phase was concentrated, 3 ml of anhydrous methanol was added to dissolve, NaBH4 (1.00 mmol) was added under ice bath conditions and stirred to react, and the reaction was monitored by TLC to be complete. The pH was adjusted to neutral with 10% HCl, and the anhydrous methanol was removed by vacuum rotary evaporation, and extracted with saturated brine and dichloromethane. The organic phase was dried over anhydrous magnesium sulfate, suction filtered, concentrated, and column chromatography was performed to obtain compound 43 as a white solid with a yield of 43.21%. 1HNMR (400MHz, Acetone) δ8.01(d,J=8.8Hz,3H),7.77(d,J=2.0Hz,1H),7.67(s,1H),7.63(d,J=8.8Hz,2H),7.37(dd,J=8 .8,2.1Hz,1H),3.92(s,2H),2.46(d,J=6.7Hz,2H),2.10(s,1H),0.93(s,1H),0.42(d,J=1.5Hz,2H),0.16–0.04(m,2H). 13 C NMR (101MHz, Acetone) δ140.36,136.43,133.96,132.26,129.84,128.74,128.63,125.24,124.82,123.16,120.31,114.92,54.20,44.49,10.61.

[0108] Synthesis of compound 44 2-(5-chloro-1-((4-chlorophenyl)sulfonyl)-1H-indol-3-yl)thiazolidine-4-carboxylic acid methyl ester

[0109] Intermediate 19-1 (782.78 mg, 2.21 mmol) was dissolved in 3 ml of anhydrous dichloromethane, and L-cysteine ​​methyl ester hydrochloride (858.25 mg, 5.00 mmol) and triethylamine (404.76 mg, 4.02 mmol) were added, and stirred at room temperature. The reaction was completed after monitoring by TLC, and anhydrous dichloromethane was removed by rotary evaporation under reduced pressure. The mixture was extracted with saturated brine and ethyl acetate, and the organic phase was dried over anhydrous magnesium sulfate. The filtrate was concentrated after suction filtration, and the target compound 44 was obtained by column chromatography. It was a white solid with a yield of 56.28%. 1 H NMR (400MHz, CDCl3) δ7.92-7.78(overlap,3H),7.77-7.62(overlap,2H),7.49-7.39(overlap,2H),7.34-7.2 8(overlap,1H),5.79(d,J=97.5Hz,1H),4.20-4.00(m,1H),3.82(s,3H),3.51-3.38(m,1H),3.15-3.07(m,1H). 13C NMR(101MHz,Acetone)δ172.62,141.38,137.09,135.08,131.62,130.88,129.6 8,126.67,126.05,125.44,121.58,121.37,115.88,64.86,64.16,52.59,38.31.

[0110] Example 45 Synthesis of Compound 45 9-Methyl-1,2,3,9-tetrahydro-4H-carbazol-4-one

[0111] Add 1,2,3,9-tetrahydro-4H-2-carbazole-4-one (100 mg, 0.53 mmol, 1 eq), sodium hydride (65 mg, 1.62 mmol, 3 eq) and 3 mL tetrahydrofuran under ice bath, stir for one hour, add iodomethane (67 μl, 1.06 mmol, 2 eq), stir at room temperature to react. TLC monitors the reaction to be complete, remove tetrahydrofuran by vacuum rotary evaporation, extract with saturated brine and ethyl acetate, dry the organic phase with anhydrous magnesium sulfate, filter and concentrate the filtrate, and obtain the target compound by column chromatography. 1 H NMR(400MHz,Chloroform-d)δ8.25(ddd,J=5.8,3.1,1.0Hz,1H),7.41–7.16(m,3H),3 .69(s,3H),2.91(t,J=6.2Hz,2H),2.56(dd,J=7.2,5.7Hz,2H),2.24(p,J=6.4Hz,2H). 13 C NMR (101MHz, Chloroform-d) δ193.73,151.80,137.39,124.76,122.95,122.55,121.64,112.68,109.08,37.85,29.88,29.79,23.31,22.18.

[0112] Example 46 Synthesis of Compound 46 9-Benzyl-1,2,3,9-tetrahydro-4H-carbazol-4-one

[0113] The synthesis process is the same as Example 45, except that methyl iodide is replaced by benzyl chloride. 1H NMR (400MHz, Chloroform-d) δ8.30 (dt, J=7.7, 1.1Hz, 1H), 7.35–7.15 (m, 6H), 7.01 (dd, J=7. 6,1.9Hz,2H),5.29(s,2H),2.84(t,J=6.2Hz,2H),2.62–2.50(m,2H),2.20(p,J=6.3Hz,2H). 13 C NMR(101MHz,Chloroform-d)δ193.95,151.80,137.16,136.06,129.05,127.90,126.11,124 .94,123.23,123.18,122.69,121.75,121.73,113.16,109.64,47.02,37.90,23.40,22.32.

[0114] Example 47 Synthesis of Compound 47 9-(Cyclopropylsulfonyl)-1,2,3,9-tetrahydro-4H-carbazol-4-one

[0115] The synthesis process is the same as Example 45, except that iodomethane is replaced by cyclopropanesulfonyl chloride. 1 H NMR(400MHz,Chloroform-d)δ8.35–8.24(m,1H),8.04–7.91(m,1H),7.43–7.30(m,2H),3.29(t,J=6.2Hz,2H),2.7 2(tt,J=8.0,4.8Hz,1H),2.61(dd,J=7.4,5.8Hz,2H),2.26(p,J=6.3Hz,2H),1.47–1.39(m,2H),1.16–1.00(m,2H). 13 C NMR(101MHz,Chloroform-d)δ195.12,151.00,136.03,125.75,125.32,124.9 4,124.89,121.99,117.67,113.54,37.92,32.34,32.25,24.55,23.28,6.24.

[0116] Example 48 Synthesis of Compound 48 9-((4-chloro-3-nitrophenyl)sulfonyl)-1,2,3,9-tetrahydro-4H-carbazol-4-one

[0117] The synthesis process is the same as Example 45, except that iodomethane is replaced by 4-chloro-3-nitrobenzenesulfonyl chloride. 1 H NMR(400MHz,Chloroform-d)δ8.40(d,J=2.3Hz,1H),8.30–8.23(m,1H),8.14–8.04(m,1H),7.93(dd,J=8.5,2.3Hz,1H ),7.68(d,J=8.5Hz,1H),7.46–7.34(m,2H),3.32(t,J=6.2Hz,2H),2.60(t,J=7.4,5.8Hz,2H),2.28(p,J=6.3Hz,2H). 13 C NMR(101MHz,Chloroform-d)δ194.73,150.20,138.19,135.60,133.92,133.80, 130.20,126.04,125.77,124.07,122.45,119.14,113.54,37.80,24.65,23.16.

[0118] Example 49 Synthesis of Compound 49 9-((4-(4-methylpiperazin-1-yl)-3-nitrophenyl)sulfonyl)-1,2,3,9-tetrahydro-4H-carbazol-4-one

[0119] Compound 48 (100 mg, 0.25 mmol, 1 eq), N-methylpiperazine (54 μL, 0.49 mmol, 2 eq), N,N-diisopropylethylamine (128 μL, 0.74 mmol, 3 eq), 1 mL of tetrahydrofuran were dissolved and stirred at room temperature. The reaction was completed by TLC monitoring, tetrahydrofuran was removed by vacuum rotary evaporation, saturated brine and ethyl acetate were extracted, the organic phase was dried over anhydrous magnesium sulfate, the filtrate was concentrated after suction filtration, and the target compound 49 was obtained by column chromatography. 1 H NMR(400MHz,Chloroform-d)δ8.32(d,J=2.4Hz,1H),8.29–8.21(m,1H),8.15–8.07(m,1H),7.77(dd,J=9.0,2.4Hz,1H),7.36(tt,J=7.3,5.7Hz,2H),7.03 (d,J=9.0Hz,1H),3.34(t,J=6.1Hz,2H),3.24–3.14(m,4H),2.58(dd,J=7.4, 5.7Hz,2H),2.50(dd,J=6.2,3.7Hz,4H),2.32(s,3H),2.26(p,J=6.3Hz,2H). 13C NMR(101MHz,Chloroform-d)δ195.00,150.64,149.05,138.48,135.68,130.88,130.85,127.06,126.75,126.73,125. 91,125.61,125.20,122.04,120.36,120.31,118.31,113.71,54.29,54.22,50.41,45.93,45.85,37.85,24.62,23.23. Application Example 1 Evaluation of Biological Activity

[0120] Experimental method: Since USP28 can cleave the C-terminus of glycine 76 of the ubiquitin molecule connected to the substrate isopeptide bond during deubiquitination, this study used the fluorescent group rhodamine (Ub1-72-Leu73-Arg74-Gly75-Gly76-Rho110, ubiquitin-rhodamine 110, Ub-Rho110) (all from Nanjing Youai Co., Ltd.) of the fluorescent peptide substrate modified with glycine 76 of ubiquitin. The enzyme activity of the recombinant protein USP28 can be quantitatively analyzed by using a microplate reader (PE Envision) and using a dichroic mirror for excitation (excitation wavelength of 485nm) and measuring the fluorescence intensity of the released rhodamine at an emission wavelength of 535nm. The activity of USP28 can be quantitatively determined and the inhibitory effect of small molecule compounds on its activity can be screened, and a USP28 inhibitor screening platform can be established.

[0121] Specific steps: First, perform a single-point 25μM screening of the inhibition rate of the compound, incubate the prepared compound with the protein USP28 (final concentration is 3.9nM) for 10 minutes, and finally add the substrate ubiquitin-rhodamine 110 (final concentration is 100nM), wait for the reaction for 5 minutes, and then quickly measure the fluorescence. If the inhibition rate is greater than 80%, we prepare the compound with a concentration gradient of 100μM, 50μM, 25μM, 12.5μM, 6.25μM, 3.12μM, 1.56μM, 0.78μM, and 0.39μM. We use a black 384-well plate (384ProxiPlate, purchased from PE Company). The final reaction volume is 20μL. 5μL of the prepared concentration gradient compound is added to each well, and the concentration becomes one-fourth of the original. In this way, the final concentration of the compound corresponding to each well is 25μM, 12.5μM, 6.25μM, 3.12μM, 1.56μM, 0.78μM, 0.39μM, 0.195μM, and 0.097μM, respectively. Finally, we use GraphPadPrism 6.0 software to calculate the inhibition rate IC 50 .

[0122] This method is also the most sensitive detection method. In addition, AZ1 was used as a positive control compound and its inhibition rate (IC 50 is 800 nM). Inhibition rate (%) = (fluorescence intensity of 100% group - fluorescence intensity of compound group) / (fluorescence intensity of 100% group - fluorescence intensity of blank group) * 100%.

[0123] Data analysis Through the above experiments, the inhibition rate of some compounds in the present invention on USP28 enzyme was tested, among which ++++: >80%; +++: 60%-80%; ++: 40%-60%; +: <40%. Table 1

[0124] The compounds with good activity of the present invention can be used as USP28 inhibitors to prevent or treat diseases related to excessive USP28 activity in humans or mammals.

Claims

1. A small molecule inhibitor of ubiquitin-specific protease 28, characterized in that: Having a molecular structure shown in general formula I, Where: The nitrogen-containing A ring represented by the dotted line is selected from one of the following substituents: X is selected from one of the following substituents: R1 is selected from the following substituents: hydrogen, or R2 is selected from the following substituents: hydrogen, or 2. The ubiquitin-specific protease 28 small molecule inhibitor according to claim 1, characterized in that: The nitrogen-containing A ring represented by the dotted line is selected from one of the following substituents: X is selected from one of the following substituents: R1 is selected from one of the following substituents: R2 is selected from one of the following substituents:

3. The ubiquitin-specific protease 28 small molecule inhibitor according to claim 1, characterized in that: Selected from the following compounds:

4. The pharmaceutical use of the small molecule inhibitor of ubiquitin-specific protease 28 according to any one of claims 1 to 3, characterized in that: The active ingredient is used for preparing medicines for preventing or treating diseases mediated by USP28.

5. The pharmaceutical use of the ubiquitin-specific protease 28 small molecule inhibitor according to claim 4, characterized in that: The compound is used for the preparation of drugs for cancer, Fanconi anemia, muscular atrophy or tumor vascular disease.

6. The pharmaceutical use of the small molecule inhibitor of ubiquitin-specific protease 28 according to claim 5, characterized in that: The cancer includes bladder cancer, pancreatic cancer, breast cancer, colon cancer, lung cancer, glioma, melanoma, head and neck cancer, gastric cancer, liver cancer, ovarian cancer, endometrial cancer, prostate cancer, thyroid cancer or kidney cancer.

7. A method for preparing a small molecule inhibitor as claimed in claim 1, characterized in that: When the nitrogen-containing A ring represented by the dotted line is 5-chloroindole, the route is as follows: X, R1 are consistent with the description of claim 1; specifically synthesized by the following steps: a) in a solvent, using compound II5-chloroindole as a starting material, formylation occurs in the presence of phosphorus oxychloride and N,N-dimethylformamide to obtain intermediate III; the solvent is: N,N-dimethylformamide; b) adding a basic compound to a solvent, and under basic conditions, reacting the formylated intermediate III obtained in step a) with benzyl chloride or sulfonyl chloride to obtain intermediate IV; the solvent is tetrahydrofuran, ethanol or dichloromethane; the basic compound is N,N-diisopropylethylamine or sodium hydride; c) In tetrahydrofuran solution, the intermediate IV obtained in step b) reacts with bromomethylbutenolide in the presence of zinc powder and saturated ammonium chloride to obtain compound I.

Images