Hetero (aromatic) ring-substituted cyclic diamine compound and use thereof in preparation of drug for treating and / or preventing tumors

Abstract

The present invention relates to a hetero (aromatic) ring-substituted cyclic diamine compound, the preparation thereof and the use thereof in the preparation of a drug for treating and / or preventing tumors. The structural formula of the compound is as shown in (I). The compound has a significant inhibitory effect on the growth of cells of tumors such as leukemia, lymphoma, breast cancer, melanoma, ovarian cancer, colorectal cancer, cervical cancer, lung cancer, prostate cancer, esophageal cancer, glioma, kidney cancer, nasopharyngeal carcinoma, liver cancer, gastric cancer and pancreatic cancer. Compared with the prior art, the compound of the present invention has a broad-spectrum anti-tumor activity, has a good effect on inhibiting tumors, and has an effect of degrading PD-L1 proteins, and is thus a PD-L1 immunomodulator.

Description

A hetero(aryl) ring-substituted cyclic diamine compound and its application in the preparation of a drug for treating and / or preventing tumors TECHNICAL FIELD

[0001] The present application relates to the technical field of medicine, in particular to a hetero(aryl) ring-substituted cyclic diamine compound and its application in the preparation of a drug for treating and / or preventing tumors. BACKGROUND

[0002] Tumors are major diseases that endanger people's lives and health, and are a global health problem. Millions of people die from tumors every year, which has a significant impact on individuals, families and society. The International Agency for Research on Cancer (IARC) of the World Health Organization released the latest global cancer burden data in 2020. In 2020, 19.3 million new cancer cases were diagnosed worldwide, and nearly 10 million people died. China has the highest number of new cancer cases and cancer deaths in the world. Due to the accelerated pace of population aging in China, changes in people's lifestyles, and an increase in cancer exposure factors, the incidence and mortality rates of various cancers in China are increasing year by year. According to the statistical data of the National Cancer Center in March 2023, lung cancer, colorectal cancer, gastric cancer, liver cancer and breast cancer are the highest incidence of cancer in China, and the incidence of various cancers is decreasing. Although China has been working hard to strengthen early prevention and treatment of tumors to alleviate the suffering of patients as much as possible, it can be predicted that tumors will continue to be a huge burden on China's health and health sector for a long time to come.

[0003] Anti-tumor treatment methods mainly include surgery, chemotherapy, radiotherapy, immunotherapy, traditional Chinese medicine treatment, etc., and chemotherapy is currently the main means of treating malignant tumors. Anti-tumor drugs are effective weapons for tumor treatment, which can save or prolong the lives of patients. The research and development of anti-tumor drugs has always been a hot and difficult topic. With the continuous development and deepening of tumor molecular biology research, the process and molecular mechanisms of the occurrence and development of malignant tumors are becoming increasingly clear, and many key molecules and genes involved in the process of tumor occurrence and development are being discovered, which has promoted the continuous development of anti-tumor drugs. New anti-tumor drugs are emerging, bringing hope to some cancer patients.

[0004] According to literature retrieval, there is no relevant report on the anti-tumor effect of hetero(aryl) ring-substituted cyclic diamine compounds so far. The present application first designs and synthesizes a hetero(aryl) ring-substituted cyclic diamine compound and explores its anti-tumor effect. SUMMARY

[0005] The purpose of the present application is to provide a new compound, a hetero(aryl) ring-substituted cyclic diamine compound, which has good anti-tumor effect, and its preparation and application in the preparation of a drug for treating and / or preventing tumors.

[0006] The object of the present application can be realized by the following technical solutions:

[0007] The present application designs a new compound, i.e. hetero(aromatic) ring substituted cyclic diamine compound, the structural formula is as follows:

[0008] In the formula:

[0009] X, Y, Z, W each independently represents carbon or nitrogen atom, and when selected from nitrogen atom, only one is nitrogen.

[0010] R1 is hydrogen, methyl, cyano, halogen, haloalkyl, primary amide, hydroxyl, mercapto, alkoxy, alkylthio, haloalkoxy, haloalkylthio, aralkyl, aryl; or combination of any two same or different groups; or combination of any three same or different groups;

[0011] R2 is methylene (-CH2-), carbonyl (-CO-).

[0012] R3 is methylene (-CH2-), 2 methylene (-CH2-CH2-), 3 methylene (-CH2-CH2-CH2-)

[0013] R4 is methylene (-CH2-), 2 methylene (-CH2-CH2-), 3 methylene (-CH2-CH2-CH2-)

[0014] R5 is benzofuran, benzopyridine, benzimidazole, benzoxazole, benzothiophene, benzopyrazole, quinoline, isoquinoline, indole, 2H-1,3-benzodioxolane 2H-chromen-2-one 1-chloro-3-methoxybenzene 2,1,3-benzothiadiazole benzene ring, pyridine ring, naphthalene ring, six-membered heterocyclic ring, benzene fused heterocyclic ring, fused heterocyclic ring.

[0015] R6 is -CONH2, -SO2NH2, 1,1,1-trifluoropropane-2-amine hydrogen, methyl, cyano, halogen, haloalkyl, hydroxyl, mercapto; or combination of any two same or different groups; or combination of any three same or different groups;

[0016] In the above chemical structure, R6 is a functional group, which can be combined with any carbon atom on the R5 ring; or R6 is absent.

[0017] Among them, halogen includes F, Cl, Br, I and the like.

[0018] Among them, haloalkyl can be monohaloalkane, dihaloalkane, polyhaloalkane.

[0019] wherein R1preferably is CF3, OCF3, SCF3.

[0020] R3preferably is -CH2-CH2-.

[0021] R4preferably is -CH2-CH2-.

[0022] R5preferably is benzofuran.

[0023] R6preferably is -CONH2, -SO2NH2, 1-methylcyclopropane-1-amine 1,1,1-trifluoropropane-2-amine

[0024] X, Y, Z, W preferably are carbon atoms.

[0025] m represents 1, 2 or 3.

[0026] n represents 0, 1, 2, 3 or 4.

[0027] Further, the hetero(aromatic) ring substituted cyclic diamine compound preferably is the following compound:

[0028] Further, the chemical structure of the above-mentioned compound is specifically as follows:

[0029] Further, the hetero(aromatic) ring substituted cyclic diamine compound includes, but is not limited to, its tautomer, meso form, racemic form, enantiomer, diastereomer or possible derivative based on its structure or mixture thereof.

[0030] The present application also provides the use of the hetero(aromatic) ring substituted cyclic diamine compound or the pharmaceutical composition comprising the compound or the derivative of the hetero(aromatic) ring substituted cyclic diamine compound in the preparation of a medicament for preventing and / or treating tumors.

[0031] Further, the hetero(aromatic) ring substituted cyclic diamine compound or the pharmaceutical composition thereof includes, but is not limited to, its pharmaceutically acceptable salt, ether, ester, prodrug, metabolite, solvate or crystal thereof.

[0032] Further, the hetero(aromatic) ring substituted cyclic diamine compound or the pharmaceutical composition thereof includes its pharmaceutically acceptable salt; preferably, the pharmaceutically acceptable salt includes, but is not limited to, hydrochloride, hydrobromide, fumarate, acetate, citrate, sulfate, methanesulfonate, formate or trifluoroacetate.

[0033] Further, the hetero (aromatic) ring-substituted cyclic diamine compound or the pharmaceutical composition thereof includes, but is not limited to, tablets, injections, capsules, oral solutions, pills, granules, powders, aerosols, patches, ointments, paints, or suppositories.

[0034] Further, the hetero (aromatic) ring-substituted cyclic diamine compound or the pharmaceutical composition thereof further includes conventional anti-tumor drugs; preferably, includes, but is not limited to, chemotherapeutic drugs, biological targeted therapeutic drugs, metabolic therapeutic drugs, or immunotherapeutic drugs.

[0035] Further, the hetero (aromatic) ring-substituted cyclic diamine compound or the pharmaceutical composition thereof is used for, but is not limited to, surgical resection, chemotherapy, or radiotherapy.

[0036] Further, the hetero (aromatic) ring-substituted cyclic diamine compound or the pharmaceutical composition thereof is used for, but is not limited to, the preparation of PD-L1 immunomodulator related drugs.

[0037] Further, the hetero (aromatic) ring-substituted cyclic diamine compound or the pharmaceutical composition thereof functions to, but is not limited to, inhibit tumor growth and / or metastasis.

[0038] Further, the hetero (aromatic) ring-substituted cyclic diamine compound or the pharmaceutical composition thereof is used for, but is not limited to, the prevention and / or treatment of leukemia, lymphoma, breast cancer, melanoma, ovarian cancer, colorectal cancer, cervical cancer, lung cancer, prostate cancer, esophageal cancer, glioma, renal cancer, nasopharyngeal cancer, liver cancer, gastric cancer, and pancreatic cancer.

[0039] Further, the prevention and / or treatment of leukemia includes, but is not limited to, the prevention and / or treatment of leukemia cells HL60.

[0040] Further, the prevention and / or treatment of lymphoma includes, but is not limited to, the prevention and / or treatment of lymphoma cells MINO.

[0041] Further, the prevention and / or treatment of breast cancer includes, but is not limited to, the prevention and / or treatment of breast cancer cells MDA-MB-468, MDA-MB-231, HS-578T, MDA-MB-436, MDA-MB-453, SPC-2, 4173, SPC-46, MDA-MB-231 LM2, MDA-MB-231 HM, HCC1954, JIMT1, T47D, MCF7.

[0042] Further, the prevention and / or treatment of melanoma includes, but is not limited to, the prevention and / or treatment of melanoma cells A375.

[0043] Further, the preventive and / or therapeutic effect on ovarian cancer includes, but is not limited to, a preventive and / or therapeutic effect on ovarian cancer cell A2780.

[0044] Further, the preventive and / or therapeutic effect on intestinal cancer includes, but is not limited to, a preventive and / or therapeutic effect on intestinal cancer cell HCT116.

[0045] Further, the preventive and / or therapeutic effect on cervical cancer includes, but is not limited to, a preventive and / or therapeutic effect on cervical cancer cell SiHa.

[0046] Further, the preventive and / or therapeutic effect on lung cancer includes, but is not limited to, a preventive and / or therapeutic effect on lung cancer cell A549.

[0047] Further, the preventive and / or therapeutic effect on prostate cancer includes, but is not limited to, a preventive and / or therapeutic effect on prostate cancer cell PC-3.

[0048] Further, the preventive and / or therapeutic effect on esophageal cancer includes, but is not limited to, a preventive and / or therapeutic effect on esophageal cancer cell EC109.

[0049] Further, the preventive and / or therapeutic effect on glioma includes, but is not limited to, a preventive and / or therapeutic effect on glioma cell U251.

[0050] Further, the preventive and / or therapeutic effect on renal clear cell adenocarcinoma includes, but is not limited to, a preventive and / or therapeutic effect on renal clear cell adenocarcinoma cell 786-O.

[0051] Further, the preventive and / or therapeutic effect on pancreatic cancer includes, but is not limited to, a preventive and / or therapeutic effect on pancreatic cancer cell Panc-1.

[0052] Further, the preventive and / or therapeutic effect on nasopharyngeal carcinoma includes, but is not limited to, a preventive and / or therapeutic effect on nasopharyngeal carcinoma cell 5-8F.

[0053] Further, the preventive and / or therapeutic effect on liver cancer includes, but is not limited to, a preventive and / or therapeutic effect on liver cancer cell SMMC-7721.

[0054] Further, the preventive and / or therapeutic effect on gastric cancer includes, but is not limited to, a preventive and / or therapeutic effect on gastric cancer cell SGC-7901.

[0055] Further, the tumor inhibition effect of JW02 on human ovarian cancer cell line A2780, human colon cancer cell line HCT-116, cervical cancer cell line SiHa, lung adenocarcinoma cell line A549, esophageal cancer cell line EC109, glioma cell line U251 and renal clear cell adenocarcinoma cell 786-O is superior to that of velaglucil and cisplatin.

[0056] The tumor inhibition effect of JW04 on human ovarian cancer cell line A2780, cervical cancer cell line SiHa, lung adenocarcinoma cell line A549 and glioma cell line U251 is superior to that of velaglucil and cisplatin.

[0057] The tumor inhibition effect of JW05 on leukemia cell HL60, melanoma cell line A375, human ovarian cancer cell line A2780, human colon cancer cell line HCT-116, cervical cancer cell line SiHa, lung adenocarcinoma cell line A549, esophageal cancer cell line EC109 and glioma cell line U251 is superior to that of velaglucil and cisplatin.

[0058] The tumor inhibition effect of JW07 on human ovarian cancer cell line A2780, human colon cancer cell line HCT-116, cervical cancer cell line SiHa, esophageal cancer cell line EC109, glioma cell line U251 and renal clear cell adenocarcinoma cell 786-O is superior to that of velaglucil and cisplatin.

[0059] The tumor inhibition effect of JW10 on human ovarian cancer cell line A2780, human colon cancer cell line HCT-116, cervical cancer cell line SiHa, lung adenocarcinoma cell line A549, esophageal cancer cell line EC109, glioma cell line U251 and renal clear cell adenocarcinoma cell 786-O is superior to that of velaglucil and cisplatin.

[0060] Compared with the prior art, the present application has the following beneficial effects:

[0061] The present application synthesizes a hetero(aryl) ring-substituted cyclic diamine compound, and explores the anti-tumor effect and the new use of the compound in the preparation of a drug for treating and / or preventing cancer.

[0062] The hetero(aryl) ring-substituted cyclic diamine compound has obvious and broad-spectrum anti-tumor activity, and has an inhibitory effect on various human tumor cells cultured in vitro, such as leukemia, lymphoma, breast cancer, melanoma, ovarian cancer, colorectal cancer, cervical cancer, lung cancer, prostate cancer, esophageal cancer, glioma, renal cancer, nasopharyngeal cancer, liver cancer, gastric cancer and pancreatic cancer.

[0063] Therefore, the hetero(aryl) ring-substituted cyclic diamine compound can be used for preparing an anti-tumor drug and preventing the occurrence and metastasis of tumors. BRIEF DESCRIPTION OF DRAWINGS

[0064] Figure 1 shows the effect of JW02 on inhibiting tumor growth in immune-deficient nude mice;

[0065] Figure 2 shows JW02 significantly inhibits the weight of tumors in immune-deficient nude mice;

[0066] Figure 3 shows JW02 significantly inhibits the volume of tumors in immune-competent mice;

[0067] Figure 4 shows JW02 significantly inhibits the weight of tumors in immune-competent mice;

[0068] Figure 5 shows the effect of hetero(aromatic) ring substituted cyclic diamine compounds on degrading PD-L1 protein. DETAILED DESCRIPTION

[0069] The principles and features of the present application are described below, and the examples are used only to explain the present application and are not intended to limit the scope of the present application. If specific conditions are not mentioned in the examples, they are carried out according to the conventional conditions or the conditions recommended by the manufacturer. If the reagents or instruments used are not mentioned by the manufacturer, they are all conventional products that can be obtained by commercial purchase.

[0070] Example 1:

[0071] 5-(4-(4-(5-carbamoyl-1H-indol-3-yl)butyl)piperazin-1-yl)benzofuran-2-carboxamide, JW01, was prepared by the following method:

[0072] In a 100 mL three-necked flask, 5-nitroindole (5 g, 35.17 mmol) was dissolved in dichloromethane (30 mL), and aluminum trichloride (9.38 g, 70.34 mmol, 3.84 mL) was added at 0 °C. The resulting mixture was stirred at 20 °C for 1 hour, and 4-chlorobutyryl chloride (7.44 g, 52.76 mmol, 5.90 mL) dissolved in dichloromethane (15 mL) was added dropwise to the reaction mixture. The resulting mixture was stirred at 20 °C for 12 hours. Thin layer chromatography (petroleum ether: ethyl acetate = 1:1) and liquid chromatography-mass spectrometry showed that the reaction was complete. The reaction mixture was added to saturated aqueous ammonium chloride solution (40 mL) at 0 °C, and extracted with dichloromethane (40 mL*3). The combined organic phase was washed with brine (30 mL*3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under vacuum to obtain the crude product. The crude product was purified by column chromatography (silica gel, petroleum ether: ethyl acetate = 1:0 to 1:1) to obtain 3-(4-chlorobutyryl)-5-nitroindole (1.4 g, 4.80 mmol, 13.65% yield) as a white solid.

[0073] Into a 100 mL three-necked flask, was taken 3-(4-chlorobutyryl)-5- cyanoindole (1.4 g, 5.68 mmol) and chlorotrimethylsilane (1.85 g, 17.03 mmol, 2.16 mL), acetonitrile (10 mL) was added and stirred to dissolve, cooled to 0 °C, sodium cyanoborohydride (713.27 mg, 11.35 mmol) was added in portions. After the addition was complete, the reaction was allowed to warm to 20 °C naturally and stirred for 3 h. The liquid chromatography-mass spectrometry showed the reaction was complete. The reaction was concentrated under reduced pressure to get the crude product. The crude product was purified by column chromatography (silica gel, petroleum ether: ethyl acetate = 1:0 to 1:1) to get 3-(4-chlorobutyl)-5-cyanoindole (970 mg, 4.00 mmol, 70.40% yield, 95.85% purity) as a white solid.

[0074] Into a 100 mL three-necked flask, was taken 5-amino benzofuran-2-carboxylic acid ethyl ester (5 g, 24.37 mmol) and dissolved in xylene (80 mL), 2-chloro-N-(2- chloroethyl)ethanamine (4.35 g, 24.37 mmol, hydrochloride salt), potassium carbonate (10.10 g, 73.10 mmol) and tetrabutylammonium bromide (1.57 g, 4.87 mmol) were added with stirring. The resulting mixture was stirred at 140 °C for 12 h. The liquid chromatography-mass spectrometry and thin layer chromatography (ethyl acetate) showed the starting material was consumed completely. The reaction was cooled and taken in distilled water (40 mL) and ethyl acetate (30 mL), then filtered to get the filtrate. The organic phase was separated and washed with brine (30 mL*3), dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under reduced pressure to get the crude product. The crude product was purified by column chromatography (silica gel, petroleum ether: ethyl acetate = 1:0 to 1:1) to get 5-(piperazin-1-yl)benzofuran-2-carboxylic acid ethyl ester (4 g, 14.58 mmol, 59.85% yield) as a white solid.

[0075] Into a 100 mL three-necked flask, was taken 5-amino benzofuran-2-carboxylic acid ethyl ester (5 g, 24.37 mmol) and dissolved in xylene (80 mL), 2-chloro-N-(2- chloroethyl)ethanamine (4.35 g, 24.37 mmol, hydrochloride salt), potassium carbonate (10.10 g, 73.10 mmol) and tetrabutylammonium bromide (1.57 g, 4.87 mmol) were added with stirring. The resulting mixture was stirred at 140 °C for 12 h. The liquid chromatography-mass spectrometry and thin layer chromatography (ethyl acetate) showed the starting material was consumed completely. The reaction was cooled and taken in distilled water (40 mL) and ethyl acetate (30 mL), then filtered to get the filtrate. The organic phase was separated and washed with brine (30 mL*3), dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under reduced pressure to get the crude product. The crude product was purified by column chromatography (silica gel, petroleum ether: ethyl acetate = 1:0 to 1:1) to get 5-(piperazin-1-yl)benzofuran-2-carboxylic acid ethyl ester (4 g, 14.58 mmol, 59.85% yield) as a white solid.

[0076] Tert-butyl 4-(2-ethoxycarbonylbenzofuran-5-yl)piperazine-1 -carboxylate (1.6 g, 4.27 mmol) was placed in a 100 mL three necked flask and dissolved with formamide (15 mL) and methanol (15 mL), sodium methoxide (1.54 g, 8.55 mmol, 30% purity) was added to the reaction at 20 °C and the internal temperature was kept below 25 °C. The resulting reaction was stirred at 20 °C for 1 h. Thin layer chromatography (petroleum ether: ethyl acetate = 1 : 1) and liquid chromatography-mass spectrometry showed the reaction was complete. Distilled water (40 mL) was added to the reaction mixture at 0 °C and filtered to collect the filter cake. The filter cake was then dried under vacuum to give tert-butyl 4-(2-carbamoylbenzofuran-5-yl)piperazine-1 -carboxylate (1.4 g, 4.05 mmol, 94.86% yield) as a white solid.

[0077] To a mixture of tert-butyl 4-(2-carbamoylbenzofuran-5-yl)piperazine-1 -carboxylate (1.4 g, 4.05 mmol) in methanol (15 mL) was added hydrogen chloride methanol (4 M, 13.12 mL) at 20 °C. The mixture was stirred at 20 °C for 1 h. Liquid chromatography-mass spectrometry showed the reaction was complete. The reaction was directly concentrated under reduced pressure to give 5-(piperazin-1 -yl)benzofuran-2-carboxamide (1.2 g, crude, hydrochloride) as a white solid.

[0078] In a 50 mL three necked flask, 5-(piperazin-1 -yl)benzofuran-2-carboxamide (120.00 mg, 424.41 μmol, hydrochloride) and 3-(4-chlorobutyl)-5-cyanoindole (100.00 mg, 429.72 umol) were dissolved in acetonitrile (3 mL), potassium iodide (11.74 g, 70.74 mmol) and diisopropylethylamine (1.48 g, 11.48 mmol, 2 mL) were added sequentially. The mixture was heated to 95 °C and stirred for 12 h. Liquid chromatography-mass spectrometry showed the reaction was complete. The reaction was also directly concentrated under reduced pressure to give the crude product. The crude product was purified by high performance liquid chromatography (column type: Welch Xtimate CI 81 150*25 mm*5 μm; mobile phase: [water (hydrochloric acid)-acetonitrile]; B%: 8%-38%, 8 min) to give 5-[4-[4-(5-cyano-1 H-indol-3-yl)butyl]piperazin-1 -yl]benzofuran-2-carboxamide (90 mg, 187.37 μmol, 82.73% yield, 99.51% purity, hydrochloride) as a yellow solid.

[0079] In a 10 mL single necked flask, 5-[4-[4-(5-cyano-1H-indol-3-yl)butyl]piperazin-1-yl]benzofuran-2-carboxamide (80 mg, 181.19 pmol) was dissolved in dimethyl sulfoxide (5 mL), potassium carbonate (6.26 mg, 45.30 pmol) and hydrogen peroxide (102.72 mg, 905.95 pmol, 87.05 pL, 30% purity) were added at 0 °C, keeping the internal temperature within 10 °C. The resulting mixture was stirred at 20 °C for 1 h, thin layer chromatography showed starting material remaining, hydrogen peroxide (236.00 mg, 2.08 mmol, 0.2 mL, 30% purity) was slowly added to the mixture at 0 °C. The resulting mixture was heated to 55 °C and stirred for 35 h. Liquid chromatography-mass spectrometry showed the reaction was complete. After cooling to room temperature, the reaction was poured into saturated aqueous sodium sulfite solution (40 mL), extracted with ethyl acetate (30 mL*3). The combined organic phase was washed with brine (30 mL*3), dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated in vacuum to give the crude product. The crude product was purified by high performance liquid chromatography (column type: Welch Xtimate C18 150*25 mm*5 pm; mobile phase: [water (hydrochloric acid)-acetonitrile]; B%: 0%-27%, 8 min) to give 5-(4-(4-(5-carbamoyl-1H-indol-3-yl)butyl)piperazin-1-yl)benzofuran-2-carboxamide (24.73 mg, 49.15 pmol, 27.12% yield, 97.43% purity, hydrochloride salt) as a white solid.

[0080] 1H NMR (400 MHz, Deuterated dimethyl sulfoxide) d = 11.07 (br. s, 1H), 10.42 (br. s, 1H), 8.18 (s, 1H), 8.06 (s, 1H), 7.85 (s, 1H), 7.66-7.64 (m, 2H), 7.53 (d, J = 9.2 Hz, 1H), 7.44 (s, 1H), 7.34 (d, J = 8.4 Hz, 1H), 7.27-7.20 (m, 3H), 7.07 (s, 1H), 3.74 (d, J = 8.4 Hz, 2H), 3.55 (s, 2H), 3.19-3.13 (m, 6H), 2.79 (t, J = 14.4 Hz, 2H), 1.79-1.71 (m, 4H).

[0081] Example 2:

[0082] 5-(4-(4-(5-(trifluoromethyl)-1H-indol-3-yl)butyl)piperazin-1-yl)benzofuran-2- carboxamide, JW02, was prepared by the following method:

[0083] Into a 100 mL three-necked flask containing trifluoromethanesulfonic acid (5 mL) was added 5-(trifluoromethyl)-indole (500 mg, 2.70 mmol) and 4-chlorobutyryl chloride (380.77 mg, 2.70 mmol, 302.20 μL) was added slowly at 0 °C. The resulting mixture was stirred at 35 °C for 1 h. Liquid chromatography-mass spectrometry showed the reaction was complete. The reaction was poured into ice-distilled water (40 mL) and a solid precipitated. The mixture was filtered and the filter cake was dried under vacuum. After drying, the filter cake was slurried with petroleum ether: ethyl acetate = 2:1 (v / v, 10 mL*3). The mixture was filtered and the filter cake was collected to give 4-chloro-1-(5-(trifluoromethyl)-1H-indol-3-yl)butan-1-one (355 mg, 1.23 mmol, 45.38% yield) as a red solid.

[0084] Into a 100 mL three-necked flask was added 4-chloro-1-[5-(trifluoromethyl)-1H- indol-3-yl]butan-1-one (355 mg, 1.23 mmol) and chlorotrimethylsilane (399.41 mg, 3.68 mmol, 466.60 μL), acetonitrile (10 mL) was added and stirred to a solution, after cooling to 0 °C, sodium cyanoborohydride (154.02 mg, 2.45 mmol) was added portionwise. The mixture was warmed to 20 °C and stirred for 3 h. Liquid chromatography-mass spectrometry showed the reaction was complete. The reaction was directly concentrated under reduced pressure to give a crude product. The crude product was poured into 10 mL distilled water and extracted with ethyl acetate (10 mL*3). The combined organic phase was washed with brine (10 mL*3), dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under vacuum. The crude product was purified by column chromatography (silica gel, petroleum ether: ethyl acetate = 1:0 to 3:1) to give 3-(4-chlorobutyl)-5-(trifluoromethyl)-indole (267 mg, 968.46 μmol, 79.03% yield) as a yellow solid.

[0085] To a 50 mL single neck flask containing 3-(4-chlorobutyl)-5-(trifluoromethyl)- 1H-indole (80 mg, 290.17 μmol) and 5-piperazin-1-ylbenzofuran-2-carboxamide (81.75 mg, 290.17 μmol, hydrochloride salt) in acetonitrile (6 mL) was added diisopropylethylamine (150.01 mg, 1.16 mmol, 202.17 μL) and potassium iodide (9.63 mg, 58.03 μmol) sequentially. The mixture was stirred at 95 °C for 24 hours. The reaction was shown to be complete by LCMS. The reaction was concentrated directly under reduced pressure to give the crude product. The residue was purified by high performance liquid chromatography (column type: Welch Xtimate C18 150*25mm*5μm; mobile phase: [water (hydrochloric acid) - acetonitrile]; B%: 20% - 50%, 8 minutes) to give 5-(4-(4-(5-(trifluoromethyl)-1H-indol-3-yl)butyl)piperazin-1-yl)benzofuran-2- carboxamide (29.72 mg, 56.32 μmol, 19.41% yield, hydrochloride salt) as a white solid.

[0086] 1H NMR (400 MHz, Deuterated dimethyl sulfoxide) δ = 11.33 (br. s, 1H), 10.29 (br. s, 1H), 8.06 (s, 1H), 7.90 (s, 1H), 7.64 (s, 1H), 7.53 (d, J = 8.8 Hz, 2H), 7.44 (s, 1H), 7.37 - 7.35 (m, 2H), 7.27 - 7.21 (m, 2H), 3.76 (d, J = 11.2 Hz, 2H), 3.58 (s, 2H), 3.19 - 3.04 (m, 6H), 2.81 (t, J = 7.2 Hz, 2H), 1.79 - 1.68 (m, 4H).

[0087] 19F NMR (400 MHz, Deuterated dimethyl sulfoxide) δ = -58.11 (s, 3F).

[0088] Example 3

[0089] 5-(4-(4-(5-cyano-2-(trimethylsilyl)-1H-pyrrolo[2,3-b]pyridin-3-yl)butyl)piperazin-1- yl)benzofuran-2-carboxamide, JW03, was prepared by the following method:

[0090] In a 10 mL single necked flask, 6-chlorohex-1-ynyl(trimethyl)silane (80 mg, 423.80 pmol) and 5-piperazin-1-ylbenzofuran-2-carboxamide (119.40 mg, 423.80 pmol, hydrochloride) were dissolved in acetonitrile (6 mL), diisopropylethylamine (219.09 mg, 1.70 mmol, 295.27 pL) and potassium iodide (7.04 mg, 42.38 pmol) were added successively. The resulting reaction was stirred at 95 °C for 24 h. LC-MS showed the reaction was completed. The reaction was directly concentrated under reduced pressure to give the crude product. The crude product was purified by high performance liquid chromatography (column type: Phenomenex luna C18 150*25mm*10 pm; mobile phase: [water (FA)-acetonitrile]; B%: 17%-47%, 58 min) to give 5-(4-(6-(trimethylsilyl)-5-hexyn-1-yl)piperazin-1-yl)benzofuran-2-carboxamide (64 mg, 143.94 pmol, 33.96% yield, 99.77% purity, formate salt) as a white solid.

[0091] Take 5-(4-(6-(trimethylsilyl)-5-hexyn-1 -yl)piperazin-1 -yl)benzofuran-2-formamide (50 mg, 125.76 pmol) and 6-amino-5-iodo-pyridine-3-carbonitrile (61.63 mg, 251.52 pmol) placed in a 10 mL single neck flask containing DMF (1 mL), add sodium carbonate (46.65 mg, 440.16 pmol,) and Pd(dppf)Cl2(10.27 mg, 12.58 pmol), lithium chloride (10.66 mg, 251.52 pmol, 5.15 pL) in turn. After nitrogen replacement for three times, the mixture was stirred at 100 °C for 12 hours. The liquid chromatography-mass spectrometry showed that the reaction was completed. After cooling, add to distilled water (20 mL), and extract with ethyl acetate (30 mL*3). The combined organic phase was washed with saturated brine (30 mL*2), dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was dissolved in tetrabutylammonium bromide (2 mL, 1 M tetrahydrofuran solution). The reaction solution was stirred at 20 °C for 1 hour, and the reaction solution was directly concentrated under reduced pressure to obtain the crude product. The crude product was purified by high performance liquid chromatography (column type: Welch Xtimate C18 150*25mm*5pm; mobile phase: [water (hydrochloric acid)-acetonitrile]; B%: 3%-33%, 8 minutes). Further purification by high performance liquid chromatography (column type: Welch Xtimate C18 150*25mm*5pm; mobile phase: [water (hydrochloric acid)-acetonitrile]; B%: 3%-33%, 8 minutes) to obtain 5-(4-(4-(5-cyano-2-(trimethylsilyl)-1 H-pyrrolo[2,3-b]pyridin-3-yl)butyl)piperazin-1 -yl)benzofuran-2-formamide (11.63 mg, 23.57 pmol, hydrochloride) as a white solid.

[0092] 1H NMR (400 MHz, Deuterated dimethyl sulfoxide) d = 12.09 (br. s, 1H), 10.35 (br. s, 1H), 8.58-8.57 (m, 2H), 8.06 (s, 1H), 7.63 (s, 1H), 7.55-7.53 (m, 2H), 7.44 (s, 1H), 7.27 (d, J = 2.0 Hz, 1H), 7.23-7.21 (m, 1H), 3.74 (s, 2H), 3.65 (d, J = 10.8 Hz, 2H), 3.190-3.10 (m, 6H), 2.77 (t, J = 7.2 Hz, 2H), 1.77-1.70 (m, 4H).

[0093] Example 4

[0094] 5-(4-(4-(5-fluoro-1H-indol-3-yl)butyl)piperazin-1-yl)benzofuran-2-carboxamide, JW04, was prepared by the following method:

[0095] To a three-necked flask containing oxalyl chloride (9.29 g, 73.20 mmol, 6.41 mL) in dichloromethane (30 mL) at -70 °C, a solution of dimethylsulfoxide (8.58 g, 109.79 mmol, 8.58 mL) in dichloromethane (10 mL) was added under nitrogen protection, keeping the internal temperature below -60 °C. The mixture was stirred at -70 °C for 0.5 h. Then a solution of 6-chlorohexan-1-ol (5 g, 36.60 mmol) in dichloromethane (10 mL) was added slowly dropwise to the mixture, keeping the internal temperature below -60 °C. The resulting mixture was stirred at -70 °C for 0.5 h. Then TEA (22.22 g, 219.59 mmol, 30.56 mL) was added slowly dropwise to the mixture at -70 °C and stirred for 0.5 h. Thin layer chromatography (petroleum ether: ethyl acetate = 1 : 1) showed that the reactants had been completely consumed and a new spot was formed. The mixture was poured into water (20 mL) and extracted with dichloromethane (10 mL*3). The combined organic phase was washed with brine (10 mL*3), dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to give 6-chlorohexanal (5.62 g, crude) as a brown oil.

[0096] To a solution of (4-fluorophenyl)hydrazine (1 g, 6.15 mmol, hydrochloride salt) in ethanol (5 mL) and distilled water (5 mL) at 20 °C, 6-chlorohexanal (993.43 mg, 7.38 mmol) was added followed by sulfuric acid (3.92 g, 8.00 mmol, 2.13 mL, 20% purity) dropwise. The resulting mixture was heated to 80 °C for 4 h. Liquid chromatography-mass spectrometry showed that the reaction was complete. The mixture was poured into saturated aqueous sodium bicarbonate solution (30 mL) at 0 °C. The aqueous phase was extracted with ethyl acetate (30 mL*3). The combined organic phase was washed with saturated brine (30 mL*3), dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated in vacuo to give the crude product. The crude product was purified by column chromatography (silica gel, petroleum ether: ethyl acetate = 1 : 0 to 3 : 1) to give 3-(4-chlorobutyl)-5-fluoro-1H-indole (365 mg, 1.40 mmol, 22.80% yield, 86.70% purity) as a yellow oil.

[0097] To a three-necked flask containing 3-(4-chlorobutyl)-5-fluoro-1H-indole (90 mg, 398.78 pmol) and 5-piperazin-1-ylbenzofuran-2-carboxamide (112.35 mg, 398.78 pmol, hydrochloride salt) in acetonitrile (6 mL) was added diisopropylethylamine (206.15 mg, 1.60 mmol, 277.83 pL) and potassium iodide (6.62 mg, 39.88 pmol) sequentially. The resulting mixture was stirred at 95 °C for 24 h. LCMS showed the reaction was completed. The reaction mixture was directly concentrated under reduced pressure to give the crude product. The crude product was purified by high performance liquid chromatography (column type: Welch Xtimate C18 150*25 mm*5 pm; mobile phase: [water (hydrochloric acid) - acetonitrile]; B%: 13% - 43%, 10 min) to give 5-(4-(4-(5-fluoro-1H-indol-3-yl)butyl)piperazin-1-yl)benzofuran-2- carboxamide (25.17 mg, 52.70 pmol, 13.21% yield, hydrochloride salt) as a white solid.

[0098] 1H NMR (400 MHz, Deuterated dimethyl sulfoxide) d = 10.92 (br. s, 1H), 10.34 (br. s, 1H), 8.05 (s, 1H), 7.63 (s, 1H), 7.53 (d, J = 9.2, 1H), 7.44 (s, 1H), 7.35 - 7.31 (m, 1H), 7.29 - 7.21 (m, 4H), 6.93 - 6.88 (m, 1H), 3.75 (d, J = 10.4 Hz, 2H), 3.55 (s, 2H), 3.19 - 3.11 (m, 6H), 2.71 (t, J = 7.6 Hz, 2H), 1.78 - 1.66 (m, 4H).

[0099] Example 5

[0100] 5-(4-(4-(5-cyano-1H-indol-3-yl)-4-oxobutyl)piperazin-1-yl)benzofuran-2- carboxamide, JW05, was prepared by the following method:

[0101] In a 50 mL three-necked flask, 3-(4-oxobutyryl)-5-cyano-indole (64.24 mg, 283.95 pmol) and 5-piperazin-1-yl-benzofuran-2-carboxamide (80 mg, 283.95 pmol, hydrochloride) were dissolved in dichloromethane (1 mL) and methanol (0.1 mL), sodium acetate (93.17 mg, 1.14 mmol) and sodium triacetoxyborohydride (150.45 mg, 709.88 pmol) were added successively. The mixture was stirred at 20 °C for 3 hours. Liquid chromatography-mass spectrometry showed that the reactants were consumed and the desired molecular weight was detected. The mixture was poured into water (30 mL), extracted with ethyl acetate (30 mL*3). The combined organic phase was washed with brine (30 mL*3), dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated in vacuum to obtain the crude product. The crude product was purified by high performance liquid chromatography (column type: Waters xbridge 150*25mm*10pm; mobile phase: [water (ammonium bicarbonate)-acetonitrile]; B%: 13%-43%, 11 min). After lyophilization, the crude product was dissolved in hydrochloric acid (0.01 M, 5 mL) and lyophilized to obtain 5-(4-(4-(5-cyano-1H-indol-3-yl)-4-oxobutyl)piperazin-1-yl)benzofuran-2- carboxamide (15.22 mg, 29.90 pmol, 10.53% yield, 96.65% purity, hydrochloride) as a white solid.

[0102] 1H NMR (400 MHz, Deuterated dimethyl sulfoxide) d = 12.39 (br. s, 1H), 8.57 (d, J = 1.2 Hz, 1H), 8.55 (s, 1H), 8.01 (s, 1H), 7.64 (d, J = 8.4 Hz, 2H), 7.58-7.56 (m, 2H), 7.45 (d, J = 4.4 Hz, 4H), 2.92 (t, J = 7.2 Hz, 2H), 2.52 (d, J = 5.2 Hz, 4H), 2.39 (t, J = 7.2 Hz, 2H), 1.87 (t, J = 7.2 Hz, 2H).

[0103] Example 6

[0104] 5-(4-(3-(5-cyano-1H-indol-3-yl)propyl)piperazin-1-yl)benzofuran-2-carboxamide, JW06, was prepared by the following method:

[0105] To a flask containing 5-cyanoindole (500 mg, 3.52 mmol) in dichloromethane (10 mL) was added aluminium trichloride (515.89 mg, 3.87 mmol, 211.43 μί) and 3-chloropropanoyl chloride (446.58 mg, 3.87 mmol, 372.44 μί) at 0 °C. The mixture was stirred at 20 °C for 16 h. LC-MS showed partial consumption of the reactant and the desired molecular weight was detected. The mixture was poured into distilled water (40 mL) at 0 °C and extracted with dichloromethane (10 mL*3). The combined organic phase was washed with brine (10 mL*3), dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated in vacuum to give the crude product. The crude product was purified by column chromatography (silica gel, petroleum ether: ethyl acetate = 1 : 1) to give 3-(3-chloropropionyl)-5-cyano-indole (355 mg, 1.53 mmol, 43.38% yield) as a white solid.

[0106] To a flask containing 3-(3-chloropropionyl)-5-cyano-indole (355 mg, 1.53 mmol) in acetonitrile (10 mL) was added trimethylsilyl chloride (497.30 mg, 4.58 mmol, 580.95 μί) and sodium cyanoborohydride (191.76 mg, 3.05 mmol) at 0 °C. The mixture was allowed to warm to 20 °C naturally and stirred for 3 h. LC-MS showed complete consumption of the reactant and the desired molecular weight was detected. The reaction was concentrated directly in vacuum to give the crude product. The crude product was purified by column chromatography (silica gel, petroleum ether: ethyl acetate = 1 :0 to 2: 1) to give 3-(3-chloropropyl)-5-cyano-indole (55 mg, 251.51 μmol, 16.48% yield, 100% purity) as a yellow oil.

[0107] To a solution of 3-(3-chloropropyl)-5-nitrile-indole (50 mg, 228.64 pmol) and 5-piperazin-1-yl-benzofuran-2-carboxamide (64.42 mg, 228.64 pmol, hydrochloride) in acetonitrile (6 mL) was added diisopropylethylamine (118.20 mg, 914.57 pmol, 159.30 pL) and potassium iodide (7.59 mg, 45.73 pmol). The mixture was stirred at 95 °C for 24 h. The reaction was directly concentrated under reduced pressure to give the crude product. The crude product was purified by high performance liquid chromatography (column type: Welch Xtimate C18 150*25 mm*5 pm; mobile phase: [water (hydrochloric acid) - acetonitrile]; B%: 6% - 36%, 8 min) to give 5-(4-(3-(5-cyano-1H-indol-3-yl)propyl)piperazin-1-yl)benzofuran-2- carboxamide (28.57 mg, 61.12 pmol, 26.73% yield, 99.25% purity, hydrochloride) as a white solid.

[0108] 1H NMR (400 MHz, Deuterated dimethyl sulfoxide) d = 11.51 (br. s, 1H), 10.06 (s, 1H), 8.15 (s, 1H), 8.06 (s, 1H), 7.63 (s, 1H), 7.55 - 7.52 (m, 2H), 7.44 - 7.42 (m, 3H), 7.27 (d, J = 2.4 Hz, 1H), 7.24 - 7.21 (m, 1H), 3.75 (d, J = 11.2 Hz, 4H), 3.60 (d, J = 11.2 Hz, 4H), 3.19 - 3.14 (m, 2H), 2.81 (t, J = 7.2 Hz, 2H), 2.13 (s, 2H).

[0109] Example 7

[0110] 3-(4-(4-(quinolin-4-yl)piperazin-1-yl)butyl)-5-cyano-indole, JW07, was prepared by the following method:

[0111] A solution of 4-chloroquinoline (500 mg, 3.06 mmol, 400.00 μL) and piperazine (2.63 g, 30.56 mmol, 10 eq) in IPA (10 mL) was stirred at 100 °C for 20 hours after which more solids precipitated. Liquid chromatography-mass spectrometry showed that the reactants were completely consumed and the desired molecular weight was detected. The mixture was filtered and the filter cake was purified by high performance liquid chromatography (column type: Phenomenex Luna C18 200*40 mm*10 μm; mobile phase: [water (TFA)-acetonitrile]; B%: 1%-10%, 10 min) to give 4-(piperazin-1-yl)quinoline (500 mg, crude) as a white solid.

[0112] To a 50 mL three-necked flask containing a solution of 4-piperazin-1-ylquinoline (50 mg, 152.76 μmol, trifluoroacetate salt) and 3-(4-chlorobutyl)-5-cyano-indole (35.55 mg, 152.76 μmol) in acetonitrile (5 mL) was added potassium iodide (5.07 mg, 30.55 μmol) and diisopropylethylamine (78.98 mg, 611.06 μmol, 106.44 μL) sequentially. The mixture was stirred at 80 °C for 24 hours. Liquid chromatography-mass spectrometry showed that the reactants were completely consumed and the desired molecular weight was detected. The reaction was directly concentrated under reduced pressure to give the crude product. The crude product was purified by high performance liquid chromatography (column type: Waters xbridge 150*25 mm*10 μm; mobile phase: [water (ammonium bicarbonate)-acetonitrile]; B%: 33%-63%, 9 min). The residue was purified by high performance liquid chromatography (column type: Welch Xtimate C18 150*25 mm*5 μm; mobile phase: [water (hydrochloric acid)-acetonitrile]; B%: 2%-32%, 8 min) to give 3-(4-(4-(quinolin-4-yl)piperazin-1-yl)butyl)-5-cyano-indole (16.29 mg, 36.28 μmol, 23.75% yield, 99.32% purity, hydrochloride salt) as a white solid.

[0113] 1H NMR (400 MHz, Deuterated dimethyl sulfoxide) δ = 11.49 (br. s, 1H), 8.84 (d, J = 6.8 Hz, 1H), 8.19 (d, J = 8.4 Hz, 2H), 8.11 (s, 1H), 8.01 (t, J = 7.2 Hz, 1H), 7.75 (t, J = 7.6 Hz, 1H), 7.51 (d, J = 8.4 Hz, 1H), 7.42-7.40 (m, 2H), 7.35 (d, J = 6.8 Hz, 1H), 4.22 (d, J = 13.2 Hz, 2H), 3.89 (s, 2H), 3.62 (d, J = 12.0 Hz, 2H), 3.35 (s, 2H), 3.22 (s, 2H), 2.79 (t, J = 7.6 Hz, 2H), 1.83-1.71 (m, 4H).

[0114] Example 8

[0115] 3-(4-(4-(2-oxo-chromen-6-yl)piperazin-1-yl)butyl)-5-cyano-indole, JW08, was prepared by the following method:

[0116] To a solution of 6-bromo-chromen-2-one (1 g, 4.44 mmol) and tert-butyl piperazine-1- carboxylate (1.24 g, 6.67 mmol) in toluene (20 mL) was added sodium tert-butoxide (854.11 mg, 8.89 mmol) and bis-tri-tert-butylphosphine palladium (681.29 mg, 1.33 mmol) at 20 °C. The reaction was purged with nitrogen and heated to 80 °C and stirred for 2 hours. Thin layer chromatography (Petroleum ether: ethyl acetate = 2:1) showed complete consumption of the reactants and formation of a new spot. The mixture was poured into water (10 mL) and extracted with ethyl acetate (10 mL*3). The combined organic phase was washed with brine (10 mL*3), dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under reduced pressure to give the crude product. The crude product was purified by column chromatography (silica gel, petroleum ether: ethyl acetate = 1:0 to 2:1) to give tert-butyl 4-(2-oxo-chromen-6-yl)piperazine-1-carboxylate (120 mg, 363.22 pmol, 8.17% yield, 100% purity) as a yellow solid.

[0117] Tert-butyl 4-(2-oxo-chromen-6-yl)piperazine-1-carboxylate (100 mg, 302.68 pmol) was added to a 50 mL vial containing hydrogen chloride (4 M solution in methanol, 5 mL) at 20 °C. The reaction was stirred at 20 °C for 1 hour. Liquid chromatography-mass spectrometry showed complete consumption of the reactants and detection of the desired molecular weight. The reaction was directly concentrated under vacuum to give 6-(piperazin-1-yl)-chromen-2-one (232 mg, crude, hydrochloride) as a white solid.

[0118] To a 50 mL three necked flask containing a solution of 6-(piperazin-1-yl)- chromen-2-one (70 mg, 262.44 μmol, hydrochloride) and 3-(4-chlorobutyl)-5- cyano-indole (61.07 mg, 262.44 μmol) in 5 mL was added diisopropylethylamine (135.67 mg, 1.05 mmol, 182.85 μL) and potassium iodide (8.71 mg, 52.49 μmol). The mixture was stirred at 95 °C for 24 h. Liquid chromatography-mass spectrometry showed that the reactants had been completely consumed and the desired molecular weight was detected. The reaction was directly concentrated under reduced pressure to give the crude product. The crude product was purified by high performance liquid chromatography (column type: Welch Xtimate C18 150*25 mm*5 μm; mobile phase: [water (hydrochloric acid)-acetonitrile]; B%: 13%-43%, 8 min) to give 3-(4-(4-(2-oxo-chromen-6-yl)piperazin-1-yl)butyl)-5-cyano-indole (20.14 mg, 41.67 μmol, 15.88% yield, 95.78% purity, hydrochloride) as a white solid.

[0119] 1H NMR (400 MHz, Deuterated dimethyl sulfoxide) δ = 11.48 (br. s, 1H), 10.66 (s, 1H), 8.10 (s, 1H), 7.97 (d, J = 9.6 Hz, 1H), 7.51 (d, J = 8.4 Hz, 1H), 7.40 (d, J = 10.8 Hz, 1H), 7.34 (s, 2H), 7.29 (s, 1H), 6.47 (d, J = 9.6 Hz, 1H), 3.80 (d, J = 10.4 Hz, 2H), 3.57 (d, J = 4.4 Hz, 4H), 3.17 - 3.11 (m, 4H), 2.78 (t, J = 7.2 Hz, 2H), 1.79 - 1.67 (m, 4H).

[0120] Example 9

[0121] 3-(4-(4-(benzo[d][1,3]dioxol-5-yl)piperazin-1-yl)butyl)-5-cyano-indole, JW09, was prepared by the following method:

[0122] To a 10 mL vial containing 1,3-benzodioxol-5-amine (500 mg, 3.65 mmol) and 2-chloro-N-(2-chloroethyl)ethanamine (780.92 mg, 4.38 mmol, hydrochloride salt) in xylene (10 mL) was added potassium carbonate (604.68 mg, 4.38 mmol) and tetrabutylammonium bromide (235.07 mg, 729.20 μmol). The mixture was heated to 140 °C and stirred for 12 h. The liquid chromatography-mass spectrometry showed the reaction was complete. The mixture was diluted with methanol (20 mL*3). The mixture was filtered to get the filtrate, which was concentrated under reduced pressure to get the crude product. The crude product was purified by reverse phase flash chromatography (column type: SepaFlash SW040 spherical C18 20-45 μm, 100 A; mobile phase: [water (0.1% trifluoroacetic acid) - acetonitrile]: 35% - 50% in 15 min) to get 1-(benzo[d][1,3]dioxol-5-yl)piperazine (370 mg, 1.79 mmol, 49.04% yield, 99.66% purity) as a yellow solid.

[0123] To a 10 mL vial containing 1-(benzo[d][1,3]dioxol-5-yl)piperazine (50 mg, 242.44 μmol) and 3-(4-chlorobutyl)-5-cyano-indole (56.42 mg, 242.44 μmol) in dimethylformamide (3 mL) was added potassium carbonate (100.52 mg, 727.31 μmol) and potassium iodide (8.05 mg, 48.49 μmol) sequentially. The mixture was stirred at 80 °C for 12 h. The liquid chromatography-mass spectrometry showed the reactants were completely consumed and the desired molecular weight was detected. The reaction was directly concentrated under reduced pressure to get the crude product. The crude product was purified by high performance liquid chromatography (column type: Waters xbridge 150*25 mm 10 μm; mobile phase: [water (ammonium bicarbonate) - acetonitrile]; B%: 34% - 64% in 11 min), further purified by high performance liquid chromatography (column type: Welch Xtimate C18 150*25 mm*5 μm; mobile phase: [water (hydrochloric acid) - acetonitrile]; B%: 15% - 45% in 8 min) to get 3-(4-(4-(benzo[d][1,3]dioxol-5-yl)piperazin-1-yl)butyl)-5-cyano-indole (38.84 mg, 86.33 μmol, 35.61% yield, 97.57% purity, hydrochloride salt) as a white solid.

[0124] 1H NMR (400 MHz, Deuterated dimethyl sulfoxide) δ = 11.47 (br. s, 1H), 10.55 (s, 1H), 8.04 (s, 1H), 7.50 (d, J = 8.4 Hz, 1H), 7.42-7.38 (m, 2H), 6.80 (d, J = 8.4 Hz, 1H), 6.75 (d, J = 2.4 Hz, 1H), 6.41-6.38 (m, 1H), 5.94 (s, 2H), 3.50 (d, J = 11.2 Hz, 4H), 3.16-3.03 (m, 6H), 2.77 (t, J = 7.2 Hz, 1H), 1.77-1.66 (m, 4H).

[0125] Example 10

[0126] 3-(4-(4-(2-chloro-6-methoxyphenyl)piperazin-1-yl)butyl)-5-cyano-indole, JW10, was prepared by the following method:

[0127] 2-chloro-6-methoxy-phenylamine (500 mg, 3.17 mmol) and 2-chloro-N-(2- chloroethyl)ethanamine (1.13 g, 6.35 mmol, hydrochloride salt) were placed in a 100 mL three-necked flask containing xylene (10 mL), tetrabutylammonium bromide (204.55 mg, 634.53 μmol) and potassium carbonate (526.17 mg, 3.81 mmol) were added sequentially. The mixture was heated to 140 °C and stirred for 12 hours. Liquid chromatography-mass spectrometry showed that the reactants had been completely consumed and the desired molecular weight was detected. The mixture was diluted with methanol (20 mL*3), filtered to obtain the filtrate, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by high performance liquid chromatography (column type: Waters xbridge 150*25mm*10μm; mobile phase: [water (ammonium bicarbonate)-acetonitrile]; B%: 14%-44%, 8 minutes) to obtain 1-(2-chloro-6-methoxyphenyl)piperazine (514 mg, 2.27 mmol, 71.46% yield) as a white solid.

[0128] Place 1-(2-chloro-6-methoxy-phenyl)piperazine (80 mg, 352.89 pmol) and 3-(4- chlorobutyl)-cyano-indole (82.12 mg, 352.89 pmol) in a 50 mL three necked flask with acetonitrile (6 mL), add diisopropylethylamine (182.43 mg, 1.41 mmol, 245.87 pL) and potassium iodide (11.72 mg, 70.58 pmol) sequentially. Stir the mixture at 95 °C for 24 hours. The liquid chromatography-mass spectrometry shows the reactants have been consumed completely, the desired molecular weight is detected. Concentrate the reaction solution directly under reduced pressure to get the crude product. Purify the crude product by high performance liquid chromatography (column type: Welch Xtimate C18 150*25mm*5pm; mobile phase: [water(acetic acid)-acetonitrile]; B%: 22%-52%, 8 minutes) to get 3-(4-(4-(2-chloro-6-methoxyphenyl)piperazin-1-yl)butyl)-5-cyano-indole (51.81 mg, 111.44 pmol, 31.58% yield, 98.82% purity, hydrochloride) as a white solid.

[0129] 1H NMR (400 MHz, Deuterated dimethyl sulfoxide) d = 11.47 (br. s, 1H), 10.18 (s, 1H), 8.10 (s, 1H), 7.51 (d, J = 8.4 Hz, 1H), 7.42-7.39 (m, 2H), 7.15 (t, J = 8 Hz, 1H), 7.06-7.01 (m, 2H), 3.82 (s, 3H), 3.59 (s, 2H), 3.46 (d, J = 10.4 Hz, 2H), 3.18 (s, 2H), 3.07-2.99 (m, 4H), 2.77 (t, J = 7.2 Hz, 2H), 1.76-1.67 (m, 4H).

[0130] Example 11

[0131] 3-(4-(4-(Benz[c][1,2,5]thiadiazol-5-yl)piperazin-1 -yl)butyl)-5-cyano-indole, JW11, was prepared by the following method:

[0132] Place 2,1,3-benzothiadiazol-5-amine (500 mg, 3.31 mmol) and 2-chloro-N-(2- chloroethyl)ethanamine (1.18 g, 6.61 mmol, hydrochloride salt) in a 100 mL three necked flask containing tulene (10 mL), add tetrabutylammonium bromide (213.22 mg, 661.42 μmol, 0.2 eq) and potassium carbonate (548.49 mg, 3.97 mmol) sequentially. Heat the mixture to 140 °C and stir for 24 hours. Liquid chromatography-mass spectrometry shows the reactants have been completely consumed, the desired molecular weight is detected. Dilute the mixture with methanol (20 mL*3), filter to get the filtrate, concentrate the filtrate under reduced pressure to get the crude product. Purify the crude product by high performance liquid chromatography (column type: Phenomenex Luna C18 200*40 mm*10 μm; mobile phase: [water (trifluoroacetic acid) - acetonitrile]; B%: 1% - 30%, 10 minutes) to get 5-(piperazin-1-yl)benzo[c][1,2,5]thiadiazole (355 mg, 1.06 mmol, 32.11% yield, trifluoroacetate salt) as a brown gum.

[0133] Place 5-piperazin-1-yl-2,1,3-benzothiadiazole (70 mg, 317.76 μmol) and 3-(4- chlorobutyl)-5-cyano-indole (73.94 mg, 317.76 μmol) in a 50 mL single necked flask containing acetonitrile (5 mL), add diisopropylethylamine (164.27 mg, 1.27 mmol, 221.39 μL) and potassium iodide (10.55 mg, 63.55 μmol) sequentially. Stir the mixture at 90 °C for 24 hours. Liquid chromatography-mass spectrometry shows the reactants have been completely consumed, the desired molecular weight is detected. Concentrate the reaction liquid directly under reduced pressure to get the crude product. Purify the crude product by high performance liquid chromatography (column type: Welch Xtimate C18 150*25 mm*5 μm; mobile phase: [water (hydrochloric acid) - acetonitrile]; B%: 16% - 46%, 8 minutes); further purify by high performance liquid chromatography (column type: Welch Xtimate C18 150*25 mm*5 μm; mobile phase: [water (hydrochloric acid) - acetonitrile]; B%: 16% - 46%, 8 minutes) to get 3-(4-(4-(benzo[c][1,2,5]thiadiazol-5-yl)piperazin-1-yl)butyl)-5-cyano-indole (14.07 mg, 30.77 μmol, 9.68% yield, 99.06% purity, hydrochloride salt) as a white solid.

[0134] 1H NMR (400 MHz, Deuterated dimethyl sulfoxide) δ = 11.45 (br. s, 1H), 10.26 (s, 1H), 8.11 (s, 1H), 7.94 (d, J = 9.6 Hz, 1H), 7.76-7.73 (m, 1H), 7.51 (d, J = 8.8 Hz, 1H), 7.43-7.39 (m, 2H), 7.28 (d, J = 2.4 Hz, 1H), 4.05 (d, J = 12.8 Hz, 2H), 3.58 (d, J = 11.6 Hz, 2H), 3.26 (s, 2H), 3.23-3.15 (m, 4H), 2.78 (t, J = 6.8 Hz, 2H), 1.78-1.69 (m, 4H).

[0135] Example 12

[0136] 2,2,2-Trifluoro-1-[5-(4-{4-[5-(trifluoromethyl)-1H-indol-3-yl]butyl}piperazin-1-yl)-1- benzofuran-2-yl]ethan-1 -amine, JW12, was prepared by the following method:

[0137] To a reaction flask was added a solution of 5-bromobenzofuran-2-carboxaldehyde (3 g, 13.33 mmol) and tetrahydrofuran (30 mL) and trimethylsilyl trifluoromethane (2.84 g, 20.00 mmol) and tetrabutylammonium fluoride (1 M, 18.00 mL) were added dropwise at 0 °C. The reaction mixture was allowed to react at 25 °C for 1 h. TLC showed the reaction was complete. The reaction was diluted with 50 mL of water and extracted with 30 mL of ethyl acetate. The organic phase was washed with 30 mL of brine and then dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give the crude product. The crude product was purified by silica gel column chromatography (PE:EA = 50:1-20:1). This gave 1-(5-bromobenzofuran-2-yl)-2,2,2-trifluoroethanol (3.29 g, 11.15 mmol, 83.64% yield) as a yellow oil.

[0138] In a reaction flask was placed 1-(5-bromobenzofuran-2-yl)-2,2,2-trifluoroethanol (560 mg, 1.90 mmol), tert-butyl piperazine-1-carboxylate (1.27 g, 5.69 mmol, hydrochloride salt), Pd2(dba)3 (173.80 mg, 189.80 μmol), cesium carbonate (3.09 g, 9.49 mmol), XPhos (271.44 mg, 569.39 μmol) and dioxane (10 mL), the reaction mixture was stirred at 80 °C for 12 h under nitrogen atmosphere. LCMS showed the reaction was completed. The reaction mixture was filtered through celite, the filter cake was rinsed with ethyl acetate, the filtrate was concentrated under reduced pressure to give a crude product. The crude product was purified by silica gel column chromatography (PE:EA = 15:1). tert-butyl 4-(2-(2,2,2-trifluoro-1-hydroxyethyl)benzofuran-5-yl)piperazine-1-carboxylate (885 mg, 2.21 mmol, 38.82% yield) was obtained as a yellow solid.

[0139] tert-butyl 4-(2-(2,2,2-trifluoro-1-hydroxyethyl)benzofuran-5-yl)piperazine-1-carboxylate

[0140] (885 mg, 2.21 mmol), IBX (1.24 g, 4.42 mmol) and EtOAc (10 mL) was placed in a reaction flask, the reaction flask was purged with nitrogen for three times, then stirred at 80 °C for 12 h under nitrogen atmosphere. LCMS showed the reaction was completed. The reaction mixture was diluted with water (20 mL), extracted with EA (20 mL*2). The combined organic layer was washed with brine (20 mL*2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a residue. The residue was purified by pre-HPLC (FA condition; column: Phenomenex luna C18 150*40mm*15μm; mobile phase: [water(FA)-ACN]; gradient: 40%-70%B, 22 min). tert-butyl 4-(2-(2,2,2-trifluoroacetyl)benzofuran-5-yl)piperazine-1-carboxylate (412 mg, 1.03 mmol, 46.79% yield) was obtained as a yellow solid.

[0141] In a reaction flask was placed tert-butyl 4-(2-(2,2,2-trifluoroacetyl)benzofuran-5-yl)piperazine-1-carboxylate (412 mg, 1.03 mmol), ACN (2 mL) and HCl / dioxane (2M, 5 mL), the reaction mixture was stirred at 25 °C for 20 min. LCMS showed the reaction was completed. The reaction mixture was concentrated under reduced pressure. The crude product 2,2,2-trifluoro-1-(5-(piperazin-1-yl)benzofuran-2-yl)ethanone (346 mg, hydrochloride salt) was obtained as a yellow oil.

[0142] In a reaction flask was added 2,2,2-trifluoro-l-(5-(piperazin-l-yl)benzofuran-2- yl)ethanone (346 mg, 1.16 mmol, hydrochloride), tetrabutylammonium bromide (1.87 g, 5.80 mmol), sodium iodide (521.66 mg, 3.48 mmol), potassium carbonate (480.98 mg, 3.48 mmol) and 3-(4-chlorobutyl)-5-(trifluoromethyl)-lH-indole (959.48 mg, 3.48 mmol) and stirred at 90 °C for 12 h. LC-MS showed the reaction was complete. The reaction mixture was diluted with water (10 mL) and extracted with dichloromethane (10 mL*3). The organic layer was washed with brine (10 mL*1), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give the crude product. The crude product was purified by pre-HPLC (formic acid condition; column: Phenomenex luna C18 150*40 mm*15 pm; mobile phase: [water (formic acid)-acetonitrile]; gradient: 25%-55% B over 15 min). 2,2,2-trifluoro-l-(5-(4-(4-(5-(trifluoromethyl)-lH-indol-3-yl)butyl)piperazin-l- yl)benzofuran-2-yl)ethanone (499 mg, 928.38 pmol, 80.03% yield, 100% purity) was obtained.

[0143] In a reaction flask was added 2,2,2-trifluoro-l-(5-(4-(4-(5-(trifluoromethyl)-lH- indol-3-yl)butyl)piperazin-l-yl)benzofuran-2-yl)ethanone (200 mg, 372.10 pmol), ethanol (5 mL), hydroxylamine hydrochloride (129.29 mg, 1.86 mmol) and triethylamine (188.26 mg, 1.86 mmol, 258.95 pL). The reaction mixture was reacted at 90 °C under nitrogen protection for 48 h. LC-MS showed the reaction was complete. The reaction mixture was diluted with 10 mL water and extracted with dichloromethane (10 mL*5), the combined organic layer was washed with brine 10 mL, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product 2,2,2-trifluoro-l-(5-(4-(4-(5-(trifluoromethyl)-lH-indol-3-yl)butyl)piperazin-l- yl)benzofuran-2-yl)ethanone oxime (205 mg) was obtained as a yellow solid and used directly in the next step.

[0144] A reaction vial was charged with 2,2,2-trifluoro-l-(5-(4-(4-(5-(trifluoromethyl)- 1H-indol-3-yl)butyl)piperazin-l-yl)benzofuran-2-yl)ethanone oxime (205 mg, 371.03 μmol), ethanol (5 mL), water (1 mL), zinc powder (280 mg, 4.28 mmol), and ammonium chloride (198.47 mg, 3.71 mmol). The reaction mixture was heated at 90 °C for 2 h under a nitrogen atmosphere. The reaction was shown to be complete by LCMS. The reaction mixture was filtered through celite and the filtrate was concentrated under reduced pressure to give a crude product. The crude product was purified by pre-HPLC (HCI conditions; Column: Agela DuraShell C18 150*25mm*5μm; Mobile Phase: [water(HCI)-ACN]; Gradient: 20%-50%B over 20 min). This gave 2,2,2-trifluoro-l-(5-(4-(4-(5-(trifluoromethyl)-lH-indol-3- yl)butyl)piperazin-l-yl)benzofuran-2-yl)ethanamine (17.04 mg, 28.75 μmol, 7.75% yield, 97.0% purity, hydrochloride salt).

[0145] 1H NMR (400 MHz, Methanol-d4) δ ppm 7.88 (s, 1H) 7.61 (d, J=9.01 Hz, 1H) 7.46 - 7.56 (m, 2H) 7.32 - 7.41 (m, 2H) 7.28 (s, 2H) 5.87 (q, J=7.09 Hz, 1H) 3.66 - 3.90 (m, 4H) 3.41 (br s, 4H) 3.27 (br d, J=8.63 Hz, 2H) 2.90 (t, J=6.88 Hz, 2H) 1.79 - 1.96 (m, 4H).

[0146] Example 13

[0147] 5-(4-{4-[5-(trifluoromethyl)-lH-indol-3-yl]butyl}piperazin-l-yl)-l-benzofuran-2- sulfonamide, JW13, was prepared by the following method:

[0148] In a reaction flask was added tert-butyl-4-(benzofuran-5-yl)piperazine-l-carboxylate (480 mg, 1.59 mmol) and 6 mL of tetrahydrofuran, the reaction flask was purged with nitrogen three times, then n-butyllithium (2.5 M, 761.99 μί) was added dropwise at -78 °C. The reaction mixture was stirred at -78 °C for 5 minutes, then t-BuONSO (257.51 mg, 1.90 mmol) was dissolved in tetrahydrofuran and added dropwise to the reaction mixture at -78 °C, the resulting reaction mixture was stirred at -78 °C for 20 minutes, then potassium hydroxide (2 M, 634.99 μί) was added to the reaction mixture, followed by warming to 60 °C, and reaction for 2 hours. LCMS showed that the reaction was complete. The reaction mixture was diluted with 20 mL of water, extracted with 10 mL of ethyl acetate three times, the organic phase was washed with 20 mL of brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give a crude product. The crude product was purified by column chromatography (Si02, petroleum ether / ethyl acetate = 6 / 1 ~ 3 / 1, TLC, petroleum ether: ethyl acetate = 2: 1, Rf= 0.18). tert-butyl-4-(2-sulfamoylbenzofuran-5-yl)piperazine-l-carboxylate (322 mg, 816.30 μmol, 51.42% yield, 96.7% purity) was obtained as a yellow solid.

[0149] In a reaction flask was added tert-butyl-4-(benzofuran-5-yl)piperazine-l-carboxylate (480 mg, 1.59 mmol) and 6 mL of tetrahydrofuran, the reaction flask was purged with nitrogen three times, then n-butyllithium (2.5 M, 761.99 μί) was added dropwise at -78 °C. The reaction mixture was stirred at -78 °C for 5 minutes, then t-BuONSO (257.51 mg, 1.90 mmol) was dissolved in tetrahydrofuran and added dropwise to the reaction mixture at -78 °C, the resulting reaction mixture was stirred at -78 °C for 20 minutes, then potassium hydroxide (2 M, 634.99 μί) was added to the reaction mixture, followed by warming to 60 °C, and reaction for 2 hours. LCMS showed that the reaction was complete. The reaction mixture was diluted with 20 mL of water, extracted with 10 mL of ethyl acetate three times, the organic phase was washed with 20 mL of brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give a crude product. The crude product was purified by column chromatography (Si02, petroleum ether / ethyl acetate = 6 / 1 ~ 3 / 1, TLC, petroleum ether: ethyl acetate = 2: 1, Rf= 0.18). tert-butyl-4-(2-sulfamoylbenzofuran-5-yl)piperazine-l-carboxylate (322 mg, 816.30 μmol, 51.42% yield, 96.7% purity) was obtained as a yellow solid.

[0150] In a reaction flask was added tert-butyl-4-(benzofuran-5-yl)piperazine-l-carboxylate (480 mg, 1.59 mmol) and 6 mL of tetrahydrofuran, the reaction flask was purged with nitrogen three times, then n-butyllithium (2.5 M, 761.99 μί) was added dropwise at -78 °C. The reaction mixture was stirred at -78 °C for 5 minutes, then t-BuONSO (257.51 mg, 1.90 mmol) was dissolved in tetrahydrofuran and added dropwise to the reaction mixture at -78 °C, the resulting reaction mixture was stirred at -78 °C for 20 minutes, then potassium hydroxide (2 M, 634.99 μί) was added to the reaction mixture, followed by warming to 60 °C, and reaction for 2 hours. LCMS showed that the reaction was complete. The reaction mixture was diluted with 20 mL of water, extracted with 10 mL of ethyl acetate three times, the organic phase was washed with 20 mL of brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give a crude product. The crude product was purified by column chromatography (Si02, petroleum ether / ethyl acetate = 6 / 1 ~ 3 / 1, TLC, petroleum ether: ethyl acetate = 2: 1, Rf= 0.18). tert-butyl-4-(2-sulfamoylbenzofuran-5-yl)piperazine-l-carboxylate (322 mg, 816.30 μmol, 51.42% yield, 96.7% purity) was obtained as a yellow solid.

[0151] To a reaction vial was added 5-(piperazin-l-yl)benzofuran-2-sulfonamide (163 mg, 579.39 μmol), acetonitrile (5 mL), potassium carbonate (240.22 mg, 1.74 mmol), tetrabutylammonium bromide (373.55 mg, 1.16 mmol), sodium iodide (260.54 mg, 1.74 mmol), and 3-(4-chlorobutyl)-5-(trifluoromethyl)-lH-indole. The reaction was stirred at 90 °C for 12 h. LCMS showed the reaction was complete. The reaction was diluted with 10 mL of water and extracted with ethyl acetate (10 mL*3), then washed with brine (10 mL*6), the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to a residue. The residue was slurried with acetonitrile for 30 min. The filter cake was then filtered and added to a reaction vial, 2 mL of methanolic hydrochloric acid (2 M) was added, stirred for 30 min, and concentrated under reduced pressure to yield 5-(4-(4-(5-(trifluoromethyl)-lH-indol-3-yl)butyl)piperazin-l-yl)benzofuran-2-sulfonamide hydrochloride (31.04 mg, 54.16 μmol, 9.35% yield, 97.2% purity, hydrochloride salt) as a off-white solid.

[0152] 1H NMR (400 MHz, Methanol-d4) δ ppm 7.88 (s, 1H) 7.50 (dd, J=17.07, 8.82 Hz, 2H) 7.35 (dd, J=8.50, 1.50 Hz, 1H) 7.24 - 7.32 (m, 4H) 3.62 - 3.82 (m, 4H) 3.04 - 3.15 (m, 2H) 2.92 (br t, J=6.50 Hz, 2H) 1.85 (br d, J=3.75 Hz, 4H).

[0153] Example 14

[0154] 5-(4-(4-(5-methoxy-lH-indol-3-yl)butyl)piperazin-l-yl)benzofuran-2-carboxamide, JW14, was prepared by the following method:

[0155] To a solution of (4-methoxyphenyl)hydrazine (1 g, 5.73 mmol, hydrochloride) in ethanol (5 mL) and distilled water (5 mL) was added 6-chlorohexanal (924.96 mg, 6.87 mmol) dropwise at 20 °C. Then to the mixture was added sulfuric acid (3.65 g, 7.44 mmol, 1.98 mL, 20% purity). The resulting mixture was heated to 80 °C for 4 hours. Liquid chromatography-mass spectrometry showed the reaction was complete. The mixture was poured into saturated aqueous sodium bicarbonate solution (30 mL) at 0 °C, extracted with ethyl acetate (30 mL*3). The combined organic phase was washed with brine (30 mL*3), dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated in vacuum to give the crude product. The crude product was purified by column chromatography (silica gel, petroleum ether: ethyl acetate = 1:0 to 3:1) to give 3-(4-chlorobutyl)-5-methoxy-1H-indole (346 mg, 1.08 mmol, 18.82% yield, 74.04% purity) as a yellow solid.

[0156] To a solution of 3-(4-chlorobutyl)-5-methoxy-1H-indole (0 mg, 378.59 μmol) and 5-piperazin-1-ylbenzofuran-2-carboxamide (106.66 mg, 378.59 μmol, hydrochloride) in a 50 mL single neck flask containing acetonitrile (6 mL) was added diisopropylethylamine (195.72 mg, 1.51 mmol, 263.77 μL) and potassium iodide (6.28 mg, 37.86 μmol) successively. The mixture was stirred at 95 °C for 24 hours. Liquid chromatography-mass spectrometry showed the reaction was complete. The reaction was directly concentrated under reduced pressure to give the crude product. The crude product was purified by high performance liquid chromatography (column type: Welch Xtimate C18 150*25mm*5μm; mobile phase: [water (hydrochloric acid)-acetonitrile]; B%: 10%-40%, 8 minutes) to give 5-(4-(4-(5-methoxy-1H-indol-3-yl)butyl)piperazin-1-yl)benzofuran-2-carboxamide (16.77 mg, 34.01 μmol, 8.98% yield, 97.96% purity, hydrochloride) as a white solid.

[0157] 1H NMR (400 MHz, Deuterated dimethyl sulfoxide) δ = 10.64 (br. s, 1H), 9.80 (s, 1H), 8.05 (s, 1H), 7.64 (s, 1H), 7.54 (d, J = 9.6 Hz, 1H), 7.44 (s, 1H), 7.27-7.26 (m, 1H), 7.24-7.21 (m, 2H), 7.11 (d, J = 2.0 Hz, 1H), 6.99 (d, J = 2.0 Hz, 1H), 6.73-6.71 (m, 1H), 3.78-3.72 (m, 5H), 3.23-3.13 (m, 6H), 3.06-3.00 (m, 2H), 2.72 (t, J = 6.8 Hz, 2H), 1.77-1.68 (m, 4H).

[0158] Example 15

[0159] 5-(4-{4-[6-(trifluoromethyl)-1H-indol-3-yl]butyl}piperazin-1-yl)-1-benzofuran-2- carboxamide, JW15, was prepared by the following method:

[0160] A mixture of 4-chlorobutyryl chloride (114.23 mg, 810.17 μmol, 90.66 μL, 1.50 eq) and titanium tetrachloride (204 mg, 1.08 mmol, 2.00 eq) in dichloromethane (3 mL) was degassed and purged with nitrogen for 15 minutes, then 6-(trifluoromethyl)-1H-indole (100.00 mg, 540 μmol, 1.00 eq) in dichloromethane was added dropwise and stirred for 2 times at 25 °C under nitrogen atmosphere. LCMS (EW52628-5-P1A) showed several new peaks on LCMS, 88.8% of the desired compound was detected. The reaction solution was quenched with 5 mL water at 0 °C, then diluted with 5 mL dichloromethane, extracted with 15 mL (5 mL*3) dichloromethane. The combined organic layer was washed with 20 mL (10 mL*2) sodium chloride, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give a residue. Purification was performed by column chromatography-thin layer chromatography (SiO2, PE:EA = 3:1). Compound 4-chloro-1-[6-(trifluoromethyl)-1H-indol-3-yl]butane (3.50 g, 11.4 mmol, yield 70.6%, purity 94.8%) was obtained as a yellow solid.

[0161] A mixture of 4-chloro-1-[6-(trifluoromethyl)-1H-indol-3-yl]butan-1-ol (1.00 g, 3.45 mmol, 1.00 eq) trimethylchlorosilane (3.00 g, 27.6 mmol, 3.51 mL, 8.00 eq) and acetonitrile (10.0 mL) and sodium cyanoborohydride (1.74 g, 27.6 mmol, 8.00 eq) was degassed and purged with nitrogen three times, then stirred at 0-25 °C under nitrogen atmosphere for 3 hours. LCMS (EW52628-7-P1A) showed several new peaks on LCMS, 83.1% of desired compound was detected. The reaction was quenched with 10.0 mL water at 0 °C, then diluted with 25 mL dichloromethane, extracted with 30.0 mL (10 mL*3) ethyl acetate. The combined organic layer was washed with 60 mL (30 mL*2) sodium chloride, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a residue. The crude product was purified by reverse phase high performance liquid chromatography (column: Phenomenex luna C18 250*50mm*10um; mobile phase: [water(FA)-ACN]; gradient: 45%-75%B over 24 minutes). Compound 3-(4-chlorobutyl)-6-(trifluoromethyl)-1H-indole (460 mg, 1.46 mmol, 42.3% yield, 87.0% purity) was obtained as a colorless oil.

[0162] (3)-4-chlorobutyl 6-1 hydrogen-indole (trifluoromethyl) (100 mg, 362 pmol, 1.00 eq), (5)-piperazine-1-acyl benzofuran-2-carboxamide (97.8 mg, 398 pmol, 1.1 eq), sodium hydride (27.2 mg, 181 pmol, 0.50 eq), potassium carbonate (100 mg, 725 pmol, 2.00 eq), tetrabutylammonium bromide (58.4 mg, 181 pmol, 0.50 eq) in acetonitrile (5.00 mL) were degassed, purged with nitrogen 3 times, stirred at 90 °C under nitrogen atmosphere for 12 hours. LCMS (EW52628-13-P1A) showed several new peaks on LCMS, 60.7% of desired compound was detected. Then diluted with water 5 mL, extracted with ethyl acetate 15 mL (5 mL*3). The combined organic layer was washed with sodium chloride 10.0 mL (5 mL*2), dried over anhydrous sodium sulfate, concentrated under reduced pressure to get a residue. The crude product was purified by reverse phase high performance liquid chromatography (column: Waters Xbridge Prep OBD C18 150*40mm*10pm; mobile phase: [water(NH4HCO3)-ACN]; gradient: 38%-68%B over 15 minutes). The crude product was fermented with methanol / hydrochloric acid (2M) at 25 °C for 60 minutes to get the target 5-[4-[4-[6-(trifluoromethyl)-1 hydrogen-indol-3-yl]butyl]piperazin-1-yl]benzofuran-2-carboxamide (30.0 mg, 56.2 pmol, 15.5% yield 15.51%, purity 97.7%, hydrochloride) as a light yellow solid.

[0163] 1H NMR (400 MHz, Deuterated chloroform) d ppm 7.72 (d, J = 8.25 Hz, 1H) 7.66 (s, 1H) 7.55 (d, J = 9.01 Hz, 1H) 7.45 (d, J = 0.63 Hz, 1H) 7.37 (d, J = 2.13 Hz, 1H) 7.24 - 7.33 (m, 3H) 3.74 - 3.81 (m, 2H) 3.67 (br d, J = 11.13 Hz, 2H) 3.35 (s, 2H) 3.14 - 3.27 (m, 4H) 2.86 - 2.95 (m, 2H) 1.86 (br d, J = 3.38 Hz, 4H).

[0164] Example 16

[0165] 5-(4-{4-[7-(trifluoromethyl)-1 hydrogen-indol-3-yl]butyl}piperazin-1-yl)-1- benzofuran-2-carboxamide, JW16, was prepared by the following method:

[0166] A mixture of 4-chlorobutyryl chloride (2.28 g, 16.20 mmol, 1.81 mL, 1.50 eq) and titanium tetrachloride (4.10 g, 21.60 mmol, 2.00 eq) in dichloromethane (100 mL) was degassed and purged with nitrogen for 15 minutes, and then a solution of 7-(trifluoromethyl)-1H-indole (2.00 g, 10.8 mmol, 1.00 eq) in dichloromethane was added dropwise. The reaction was stirred at 25 °C under nitrogen atmosphere for 2 times. LCMS (EW52628-6-P1A) showed several new peaks on LCMS, and 98.5% of the target compound was detected. The reaction was quenched with 5 mL of water at 0 °C, and then diluted with 5 mL of dichloromethane, and extracted with 15 mL (5 mL*3) of dichloromethane. The combined organic layer was washed with 20 mL (10 mL*2) of sodium chloride, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give a residue. The compound was purified by column chromatography-thin layer chromatography (SiO2, PE:EA = 3:1). Compound (4-chloro-1-(7)-trifluoromethyl-1H-indol-3-yl)butan-1-1 (3.00 g 10.24 mmol, yield 94.82%, purity 98.9%) was a yellow solid.

[0167] A mixture of 4-chloro-1-[7-(trifluoromethyl)-1H-indol-3-yl]butan-1-1 (1.00 g, 3.45 mmol, 1.00 eq), trimethylsilyl chloride (3.00 g, 27.6 mmol, 3.50 mL, 8.00 eq), acetonitrile (10.0 mL), and sodium cyanoborohydride (1.74 g, 27.6 mmol, 8.00 eq) was degassed and purged with nitrogen for 3 times, and then stirred at 0-25 °C under nitrogen atmosphere for 3 hours. LCMS (EW52628-8-P1A) showed several new peaks on LCMS, and 95.9% of the target compound was detected. The reaction was quenched with 10.0 mL of water at 0 °C, and then diluted with 25 mL of dichloromethane, and extracted with 30.0 mL (10 mL*3) of ethyl acetate. The combined organic layer was washed with 60 mL (30 mL*2) of sodium chloride, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give a residue. The crude product was purified by reverse phase high performance liquid chromatography (column: Phenomenex luna C18 250*50 mm*10 μm; mobile phase: [water (FA)-ACN]; gradient: 45%-75% B, over 24 minutes). Compound 3-(4-chlorobutyl)-7-(trifluoromethyl)-1H-indole (820 mg, 2.97 mmol, yield 85.9%, purity 99.8%) was a colorless oil.

[0168] (3)-4-chlorobutyl 7-1 hydrogen-indole(trifluoromethyl) (100 mg, 362 pmol, 1.00 eq), (5)-piperazine-1-acyl benzofuran-2-carboxamide (97.8 mg, 398 pmol, 1.1 eq), sodium hydride (27.2 mg, 181 pmol, 0.50 eq), potassium carbonate (100 mg, 725 pmol, 2.00 eq), tetrabutylammonium bromide (58.4 mg, 181 pmol, 0.50 eq) in acetonitrile (5.00 mL) were degassed, nitrogen purged 3 times, stirred at 90 °C under nitrogen atmosphere for 12 hours. LCMS (EW52628-14-P1C) appeared several new peaks on LCMS, 86.2% of the target compound was detected. Then diluted the ethyl acetate with water 5 mL, extracted with ethyl acetate 15 ml (5 mL*3). The combined organic layer was washed with sodium chloride 10.0 mL (5 mL*2), dried over anhydrous sodium sulfate, concentrated under reduced pressure to get a residue. The crude product was purified by reverse phase high performance liquid chromatography (column: Phenomenex luna C18 150*25mm*10pm, mobile phase: [water(HCl)-ACN], gradient: 18%-48%B, 10 minutes). The target 5-[4-[4-[7-(trifluoromethyl)-1 hydrogen-indol-3-yl]butyl]piperazin-1-yl]benzofuran-2-carboxamide (25.0 mg, 47.4 pmol, yield 13.1%, purity 98.8%, hydrochloride) was an off-white solid.

[0169] 1H NMR (400 MHz, Methanol-d4) d ppm 7.83 (d, J = 7.88 Hz, 1H) 7.53 (d, J = 9.13 Hz, 1H) 7.44 (s, 1H) 7.40 (d, J = 7.38 Hz, 1H) 7.33 (d, J = 2.38 Hz, 1H) 7.27 (m, 1H) 7.22 (s, 1H) 7.11 - 7.17 (m, 1H) 3.72 - 3.82 (m, 2H) 3.61 - 3.70 (m, 2H) 3.22 - 3.29 (m, 4H) 3.06 - 3.18 (m, 2H) 2.87 - 2.97 (m, 2H) 1.80 - 1.90 (m, 4H).

[0170] Example 17

[0171] 5-(4-{4-[4-(trifluoromethyl)-1 hydrogen-indol-3-yl]butyl}piperazin-1-yl)-1- benzofuran-2-carboxamide, JW17, was prepared by the following method:

[0172] To a suspension of 4-chlorobutyryl chloride (2.28 g, 16.2 mmol, 1.81 mL, 1.50 eq) in dichloromethane (80.0 mL) was added titanium tetrachloride (4.10 g, 21.6 mmol, 2.00 eq) at -60 °C. After stirring for a few minutes, a solution of 4-trifluoromethylindole (2.00 g, 10.8 mmol, 1.00 eq) in dichloromethane (80.0 mL) was added dropwise. The reaction was then returned to -60 °C and continued for 2 hours. Thin layer chromatography (PE:EA = 1:1 Rf = 0.45) showed that 4-trifluoromethylindole was still present and a new spot of greater polarity was detected. The reaction mixture was quenched with water 100 mL at 0 °C and extracted with dichloromethane (60 mL*3). The combined organic layers were washed with brine (90 mL*3), dried over anhydrous sodium sulfate and concentrated under reduced pressure to give a residue. The crude product was purified using petroleum ether: ethyl acetate = 5:1, slurry at 25 °C to give 4-chloro-1-(4)-trifluoromethyl 1-hydro-indol-3-yl)butane-1-1 (783 mg, 2.58 mmol, 23.8% yield, 95.4% purity) as an off-white solid.

[0173] To 4-chloro-1-(4)-trifluoromethyl 1-hydro-indol-3-yl)butane-1-1 (733 mg, 2.53 mmol, 1.00 eq) in 8.00 mL of acetonitrile was added sodium cyanoborohydride (1.27 g, 20.2 mmol, 8.00 eq) and trimethylsilyl chloride (2.20 g, 20.2 mmol, 2.57 mL, 8.00 eq) at 0 °C. The reaction was allowed to warm to 25 °C and stirred at 25 °C for 12 hours. LCMS (EW52633-12-P1A1, 254 nm) showed that 4-chloro-1-(4)-trifluoromethyl 1-hydro-indol-3-yl)butane-1-1 was completely consumed and 48.8% of the desired mass was detected. The reaction mixture was added to water 10 mL and ethyl acetate 10 mL. The organic phase was separated, washed with brine (10 mL*1), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure to give a crude product. The crude product was purified by reverse phase high performance liquid chromatography (column: Phenomenex luna C18 150*40 mm*15 μm, mobile phase: [water (FA)-ACN], gradient: 55%-85% B over 15 minutes). 3-4-chlorobutyl-4-1h-indole(trifluoromethyl) (180 mg, 645 μmol, 25.4% yield, 98.8% purity) was an off-white solid.

[0174] To a solution of 3-(4-chlorobutyl)-4-(1H-indol-3-yl)-1H-indole (130 mg, 471 μmol, 1.00 eq) in acetonitrile (5 mL) was added 1-(2-aminocarbonylbenzofuran-5-yl)piperazine (115 mg, 471 μmol, 1.00 eq), sodium iodide (35.3 mg, 236 μmol, 0.50 eq), potassium carbonate (162 mg, 1.18 mmol, 2.50 eq), and tetrabutylammonium bromide (76.0 mg, 236 μmol, 0.50 eq). The reaction mixture was purged with nitrogen three times and then heated to 90 °C under nitrogen atmosphere for 4 h. LCMS (EW52633-16-P1A) showed 20.7% of 3-(4-chlorobutyl)-4-(1H-indol-3-yl)-1H-indole remained. Several new peaks appeared on LCMS and 56.4% of the desired compound was detected. The reaction mixture was diluted with saturated aqueous sodium bicarbonate solution (10 mL) and ethyl acetate (30 mL), the organic phase was separated, washed with brine (10 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give a crude product. The residue was purified by preparative column chromatography (SiO2, dichloromethane:methanol = 8:1), then the pure product was slurried in acetonitrile (5 volumes) at 25 °C for 60 min, filtered, and finally 2 mol of hydrochloric acid in methanol was added to the pure product at 25 °C, stirred for 2 h to give 5-(4-(4-(4-(trifluoromethyl)-1H-indol-3-yl)butyl)piperazin-1-yl)benzofuran-2-carboxamide (25 mg, 47.5 μmol, 10.0% yield, hydrochloride salt) as an off-white solid.

[0175] 1H NMR (400 MHz, Deuterated dimethyl sulfoxide) δ 1.66 - 1.77 (m, 2H) 1.83 - 1.95 (m, 2H) 2.77 (br t, J = 7.38 Hz, 2H) 3.14 - 3.24 (m, 6H) 3.60 (br d, J = 6.50 Hz, 2H) 3.69 - 3.84 (m, 2H) 7.16 - 7.25 (m, 2H) 7.28 (d, J = 2.25 Hz, 1H) 7.40 (d, J = 7.38 Hz, 1H) 7.45 (s, 2H) 7.54 (d, J = 9.01 Hz, 1H) 7.64 (br s, 1H) 7.69 (d, J = 8.13 Hz, 1H) 8.08 (br s, 1H) 10.73 - 10.90 (m, 1H) 11.60 (br s, 1H).

[0176] Example 18

[0177] 5-{4-[4-(5-chloro-1H-indol-3-yl)butyl]piperazin-1-yl}-1-benzofuran-2-carboxamide, JW18, was prepared by the following method:

[0178] To a suspension of aluminium trichloride (3.52 g, 26.4 mmol, 1.44 mL, 2.00 eq) in dichloromethane (50 mL) was added 4-chlorobutyryl chloride (2.79 g, 19.8 mmol, 2.21 mL, 1.50 eq) at 0 °C. After stirring for a few minutes, a solution of 5-chloroindole (2 g, 13.19 mmol, 1.00 eq) in dichloromethane (50 mL) was added gradually. The reaction was then allowed to return to 25 °C and continue for 2 hours. Thin layer chromatography (PE:EA = 1:1 Rf = 0.45) showed that 5-chloroindole was still present and a new spot of greater polarity was detected. The reaction mixture was quenched with water 500 mL at 0 °C and then extracted with ethyl acetate (300 mL*3). The organic phase was separated, washed with brine (450 mL*3), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a crude product. The crude product was slurried in methanol (5 volumes) at 25 °C for 60 minutes to give 4-chloro-1-5-chloro-1H-indol-3-ylbutan-1-1 (1.82 g, 6.64 mmol, 50.3%, purity 93.5%) as an off-white solid. (1H NMR: EW52633-7-P1DJ, LCMS: EW52633-7-P1DJ, HPLC: EW52633-7-P1DJ1)

[0179] To a solution of 4-chloro-1-5-chloro-1H-indol-3-ylbutan-1-1 (500 mg, 1.95 mmol, 1.00 eq) and sodium cyanoborohydride (981 mg, 15.6 mmol, 8.00 eq) in acetonitrile (5.00 mL) was added trimethylsilylchloride (1.70 g, 15.6 mmol, 1.98 mL, 8.00 eq) at 0 °C and stirred for 4 hours. Thin layer chromatography (PE:EA = 2:1, Rf = 0.58) showed that 4-chloro-1-5-chloro-1H-indol-3-ylbutan-1-1 remained and many new spots were formed. Thin layer chromatography showed that the reaction was very messy. The reaction mixture was added to water 20 mL and extracted with ethyl acetate (10 mL*3). The organic phase was separated, washed with brine (15 mL*1), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a residue. The crude product was purified by reverse phase high performance liquid chromatography (column: Phenomenex luna C18 150*40 mm*15 μm, mobile phase: [water (FA)-ACN], gradient: 55%-85% B over 15 minutes). (High performance liquid chromatography: EW52633-11-P1ZB). 5-chloro-3-4-chlorobutyl 1H-indole (200 mg, 733 μmol, 37.5% yield, 88.8% purity) was a yellow oil.

[0180] To a solution of 5-chloro-3-4-chlorobutyl 1-h-indole (150 mg, 619 pmol, 1.00 eq), 1-(2-aminocarbonylbenzofuran-5-yl)piperazine (167 mg, 681 pmol, 1.10 eq), sodium iodide (46.4 mg, 310 pmol, 0.50 eq), potassium carbonate (214 mg, 1.55 mmol, 2.50 eq), tetrabutylammonium bromide (99.8 mg, 310 pmol, 0.50 eq) in acetonitrile, purged with nitrogen for 3 times, then warmed to 90 °C, stirred for 16 hours under nitrogen atmosphere. LCMS (EW52633-14-P1A) showed 5-chloro-3-4-chlorobutyl 1-h-indole was consumed completely, 63.8% of desired mass was detected. The reaction mixture was filtered, and the filtrate was concentrated. The crude product was purified by reverse phase high performance liquid chromatography (column: Phenomenex luna C18 150*25mm*10pm, mobile phase: [water(HCl)-ACN], gradient: 15%-45%B, 10 minutes), but thin layer chromatography showed the product was messy, then diluted with sodium bicarbonate aqueous solution 10 mL, extracted with dichloromethane:methanol = 10:1 (20 mL*2). The combined organic layer was washed with brine (10 mL*1), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by preparative chromatography column (SiO2, DCM:MeOH = 8:1). Then the product was mixed with hydrochloric acid in methanol solution (2.00 M) and stirred at 25 °C for 2 hours to give 5-[4-[4-(5-chloro-1-h-indol-3-yl)butyl]piperazin-1-yl]benzofuran-2-carboxamide (20.0 mg, 39.8 pmol, 6.43% yield, 97.1% purity, hydrochloride) as an off-white solid.

[0181] 1H NMR (400 MHz, Deuterated dimethyl sulfoxide) d 1.60 - 1.74 (m, 2H) 1.74 - 1.87 (m, 2H) 2.66 - 2.78 (m, 2H) 3.07 - 3.25 (m, 6H) 3.51 - 3.60 (m, 2H) 3.73 - 3.75 (m, 2H) 7.06 (dd, J = 8.57, 1.94 Hz, 1H) 7.18 - 7.31 (m, 3H) 7.36 (d, J = 8.63 Hz, 1H) 7.45 (s, 1H) 7.51 - 7.59 (m, 2H) 7.63 (br s, 1H) 8.07 (br s, 1H) 10.62 - 10.78 (m, 1H) 11.00 - 11.13 (m, 1H).

[0182] Example 19

[0183] 5-[4-(4-{1 Hydro-pyrrolo[3,2-c]pyridin-3-yl}butyl)piperazin-1-yl]-1- benzofuran-2-carboxamide, JW19, was prepared by the following method:

[0184] Titanium tetrachloride (32.1 g, 169 mmol, 2.00 eq) was added to a solution of dichloromethane (100 mL) and 4-chlorobutyryl chloride (14.3 g, 102 mmol, 11.4 mL, 1.20 eq) was added dropwise over 30 minutes at 0 °C, followed by the dropwise addition of 1H-pyrrolo[3,2-c]pyridine (10.0 g, 84.7 mmol, 1.00 eq) at 0 °C. The resulting mixture was stirred at 25 °C for 12 hours. Thin layer chromatography (dichloromethane:methanol = 10:1, Rf = 0.53) indicated that the starting material was consumed and several new spots were formed. The reaction mixture was quenched by the addition of sodium bicarbonate at 0 °C and extracted with dichloromethane (100 mL*2). The organic layer was washed with saturated aqueous sodium chloride (100 mL*1), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a solid. The solid was purified by column chromatography (SiO2, dichloromethane:methanol = 30:1~15:1). 4-chloro-1-(1 Hydro-pyrrolo[3,2-c]pyridin-3-yl)butan-1-one (2.20 g, 9.88 mmol, 11.6% yield) was obtained as a white solid.

[0185] Sodium borohydride (5.67 g, 150 mmol, 15.2 eq) was added to trifluoroacetic acid (60.0 mL) and stirred at 0 °C for 10 minutes. 4-chloro-1-(1 Hydro-pyrrolo[3,2-c]pyridin-3-yl)butan-1-one (2.20 g, 9.88 mmol, 1.00 eq) was dissolved in dichloromethane (22.0 mL) and added dropwise to the reaction mixture at 0 °C. The resulting mixture was stirred at 25 °C for 12 hours. Liquid chromatography-mass spectrometry showed that the reaction was complete and the product was detected. The reaction mixture was quenched by the addition of saturated aqueous ammonium chloride 100 mL at 0 °C and then diluted with dichloromethane 50.0 mL and extracted with dichloromethane (50.0 mL*2). The organic layer was washed with saturated aqueous sodium chloride (100 mL), dried over anhydrous sodium sulfate, filtered and concentrated to give a solid residue. The solid residue was purified by high performance liquid chromatography (column: Welch Ultimate XB-SiOH 250*70*10 μm; mobile phase: [hexane-ethane]; gradient: 5%-45% B for 19 minutes). 3-(4-chlorobutyl)-1 Hydro-pyrrolo[3,2-c]pyridine (550 mg, 2.11 mmol, 21.3% yield, 80% purity) was obtained as a yellow oil.

[0186] Compound 3-(4-chlorobutyl)-1H-pyrrolo[3,2-c]pyridine (550 mg, 2.64 mmol, 1.00 eq), 5-piperazin-1-ylbenzofuran-2-carboxamide, sodium iodide (790 mg, 5.27 mmol, 2.00 eq) and potassium carbonate (1.09 g, 7.91 mmol, 3.00 eq) were added sequentially to acetonitrile (10.0 mL). Stirring was continued at 90 °C for 48 h. Liquid chromatography-mass spectrometry showed that the content of starting material in the reaction solution was ~23.6% and the content of product was ~25.3%. The reaction mixture was concentrated under reduced pressure to give a residue. Pre-high performance liquid chromatography (column: Waters Xbridge Prep OBD C18 150*40 mm*10 pm; mobile phase: [water (ammonium bicarbonate)-acetonitrile]; gradient: 10%-40% B for 15 min) was used for purification. 5-[4-[4-(1H-pyrrolo[3,2-c]pyridin-3-yl)butyl]piperazin-1-yl]benzofuran-2- carboxamide (25.0 mg, 59.1 pmol, 2.24% yield, 98.7% purity) was obtained as a yellow solid.

[0187] Compound 5-[4-[4-(1H-pyrrolo[3,2-c]pyridin-3-yl)butyl]piperazin-1-yl]benzofuran-2- carboxamide (25.0 mg, 59.88 pmol, 1.00 eq) was added to hydrochloric acid / methanol (2.00 M, 0.5 mL, 16.70 eq). Stirring was continued at 25 °C for 2 h. Thin layer chromatography (dichloromethane:methanol = 1:1, Rf = 0.14) showed that the starting material was consumed and a new spot was formed. The reaction mixture was concentrated under reduced pressure to give a residue. Pre-thin layer chromatography (dichloromethane:methanol = 1:1) was used for purification. Compound 5-[4-[4-(1H-pyrrolo[3,2-c]pyridin-3-yl)butyl]piperazin-1-yl]benzofuran-2- carboxamide (21.0 mg, 48.5 pmol, 73.3% yield, hydrochloride salt) was obtained as a white solid.

[0188] 1H NMR (400 MHz, Methanol-d4) d 1.86 - 2.03 (m, 4H) 2.94 - 3.05 (m, 2H) 3.23 (br d, J = 12.51 Hz, 2H) 3.33 (s, 2H) 3.39 - 3.50 (m, 1H) 3.67 - 3.82 (m, 4H) 7.28 (dd, J = 9.07, 2.31 Hz, 1H) 7.34 (d, J = 2.00 Hz, 1H) 7.44 (d, J = 0.75 Hz, 1H) 7.53 (d, J = 9.01 Hz, 1H) 7.70 (s, 1H) 7.89 (d, J = 6.75 Hz, 1H) 8.30 (d, J = 6.75 Hz, 1H) 9.23 (s, 1H) 12.38 (br s, 1H).

[0189] Performance detection

[0190] The performance of the hetero(aryl) ring-substituted cyclic diamines prepared in the above examples was detected.

[0191] (1) Detection of the effect of each compound on inhibiting the growth of two tumor cell lines

[0192] The cytotoxicity of 19 hetero(aryl) ring-substituted cyclic diamines and cisplatin on human breast cancer MDA-MB-231 LM2 cell line and lung adenocarcinoma A549 cell line was compared by CCK8 colorimetry.

[0193] The 19 hetero(aryl) ring-substituted cyclic diamines or cisplatin were diluted to concentrations of 0.01, 0.1, 0.5, 1, 10, 20, 40, and 80 μM, respectively. Breast cancer MDA-MB-231 LM2 cells or lung adenocarcinoma A549 cells were inoculated in 96-well plates at a density of 8000 cells per well. After 24 h, the original culture medium was discarded, and 100 μL of the diluted drug solution was added to each well, with 4 replicate wells in each group. After 48 h of treatment, 10 μL of CCK8 was added to each well for incubation for 3 h. The OD value at A450 was detected by a multifunctional enzyme marker, and the inhibition rate was calculated.

[0194] The results showed that most of the hetero(aryl) ring-substituted cyclic diamines had inhibitory effects on the two tumor cell lines. The IC 50 of some compounds (such as JW02, JW05, JW10, JW12, JW13, JW17, and JW18) was significantly lower than that of cisplatin. (See Table 1 for details)

[0195] Table 1 IC of hetero(aryl) ring-substituted cyclic diamines and cisplatin on two tumor cell lines after 48 h of treatment 50

[0196] (2) Detection of the effect of each hetero(aryl) ring-substituted cyclic diamine on inhibiting the growth of multiple tumor cell lines

[0197] CCK8 colorimetric method was used to detect the cytotoxicity of hetero (aromatic) ring substituted cyclic diamines (JW02, JW04, JW05, JW07, JW10), verazocine and cisplatin on human leukemia cell line HL60, lymphoma cell line MINO, breast cancer cell line MDA-MB-231 LM2, melanoma cell line A375, human ovarian cancer cell line A2780, human colon cancer cell line HCT-116, cervical cancer cell line SiHa, lung adenocarcinoma cell line A549, prostate cancer cell line PC-3, esophageal cancer cell line EC109, glioma cell line U251, renal clear cell adenocarcinoma cell 786-O, nasopharyngeal carcinoma cell line 5-8F, hepatocarcinoma cell line SMMC-7721, gastric adenocarcinoma cell line SGC-7901, and pancreatic cancer cell line Panc-1.

[0198] Five hetero (aromatic) ring substituted cyclic diamines (JW02, JW04, JW05, JW07, JW10) or cisplatin or verazocine were selected and diluted to 0, 0.658, 1.975, 5.925, 17.777, 53.333, 160 μM with a 3-fold gradient, with cisplatin and verazocine as control drugs. Various cells were inoculated in 96-well plates at a density of 8000 cells per well, and after 24 h of inoculation, the culture medium in the original well was discarded, 100 μL of diluted drug solution was added to each well, and 4 replicate wells were set up for each group. After 48 h of treatment, 10 μL of CCK8 was added to each well for 3 h of incubation, and then the OD value at A450 was detected using a multifunctional enzyme marker, and the inhibition rate was calculated.

[0199] Table 2 IC of partial hetero (aromatic) ring substituted cyclic diamines on various tumor cell lines after 48 h of treatment 50

[0200] The results showed that the five hetero (aromatic) ring substituted cyclic diamines and cisplatin and verazocine had inhibitory effects on various tumor cells.

[0201] In human ovarian cancer cell line A2780, human colon cancer cell line HCT-116, cervical cancer cell line SiHa, lung adenocarcinoma cell line A549, esophageal cancer cell line EC109, glioma cell line U251, and renal clear cell adenocarcinoma cell 786-O, JW02 had a lower IC 50; and in lymphoma cell line MINO, melanoma cell line A375, human ovarian cancer cell line A2780, human colon cancer cell line HCT-116, cervical cancer cell line SiHa, lung adenocarcinoma cell line A549, prostate cancer cell line PC-3, esophageal cancer cell line EC109, glioma cell line U251, renal clear cell adenocarcinoma cell 786-O, nasopharyngeal carcinoma cell line 5-8F, and gastric adenocarcinoma cell line SGC-7901, JW02 has lower IC 50 ; and in human ovarian cancer cell line A2780, human colon cancer cell line HCT-116, cervical cancer cell line SiHa, lung adenocarcinoma cell line A549, esophageal cancer cell line EC109, glioma cell line U251, and renal clear cell adenocarcinoma cell 786-O, JW02 has lower IC 50 than verapamil and cisplatin.

[0202] JW04 has killing effect on different types of tumor cells, and its IC 50 is not much different from that of verapamil and cisplatin. In human ovarian cancer cell line A2780, cervical cancer cell line SiHa, lung adenocarcinoma cell line A549, and glioma cell line U251, JW04 has lower IC 50 than verapamil and cisplatin.

[0203] In leukemia cell line HL60, breast cancer cell line MDA-MB-231LM2, melanoma cell line A375, human ovarian cancer cell line A2780, human colon cancer cell line HCT-116, cervical cancer cell line SiHa, lung adenocarcinoma cell line A549, esophageal cancer cell line EC109, and glioma cell line U251, JW05 has lower IC 50 than cisplatin; and in all cell lines except hepatocarcinoma cell line SMMC-7721 and pancreatic cancer cell line Panc-1, JW05 has lower IC 50 than verapamil. In leukemia cell line HL60, melanoma cell line A375, human ovarian cancer cell line A2780, human colon cancer cell line HCT-116, cervical cancer cell line SiHa, lung adenocarcinoma cell line A549, esophageal cancer cell line EC109, and glioma cell line U251, JW05 has lower IC 50 than verapamil and cisplatin.

[0204] In breast cancer cell line MDA-MB-231LM2, human ovarian cancer cell line A2780, human colon cancer cell line HCT-116, cervical cancer cell line SiHa, esophageal cancer cell line EC109, glioma cell line U251, renal clear cell adenocarcinoma cell 786-O, and pancreatic cancer cell line Panc-1, JW07 has lower IC 50, and has better inhibitory effect on tumor; in lymphoma cell line MINO, melanoma cell line A375, cervical cancer cell line SiHa, lung adenocarcinoma cell line A549, prostate cancer cell line PC-3, esophageal cancer cell line EC109, glioma cell line U251, renal clear cell adenocarcinoma cell 786-O, and nasopharyngeal carcinoma cell line 5-8F, JW07 has lower IC 50 . Among them, JW07 has lower IC 50 than that of verapamil and cisplatin in human ovarian cancer cell line A2780, human colon cancer cell line HCT-116, cervical cancer cell line SiHa, esophageal cancer cell line EC109, glioma cell line U251, and renal clear cell adenocarcinoma cell 786-O.

[0205] In breast cancer cell line MDA-MB-231LM2, human ovarian cancer cell line A2780, human colon cancer cell line HCT-116, cervical cancer cell line SiHa, lung adenocarcinoma cell line A549, esophageal cancer cell line EC109, glioma cell line U251, renal clear cell adenocarcinoma cell 786-O, and pancreatic cancer cell line Panc-1, JW10 has lower IC 50 than that of cisplatin. Except for MDA-MB-231LM2, JW10 has lower IC 50 than that of verapamil in all the remaining cell lines. 50 Among them, JW10 has lower IC

[0206] than that of verapamil and cisplatin in human ovarian cancer cell line A2780, human colon cancer cell line HCT-116, cervical cancer cell line SiHa, lung adenocarcinoma cell line A549, esophageal cancer cell line EC109, glioma cell line U251, and renal clear cell adenocarcinoma cell 786-O.

[0207] (3) Inhibitory effect of hetero(aryl) ring-substituted cyclic diamine compounds on the growth of 13 breast tumor cells

[0208] The cytotoxicity of hetero(aromatic) ring-substituted cyclic diamines (JW02, JW04, JW05, JW07, JW10), verapamil and cisplatin on triple-negative breast cancer (TNBC) cell lines (MDA-MB-468, MDA-MB-231, Hs578T, MDA-MB-436, MDA-MB-453, SPC-2, MDA-MB-231 4173, SPC46, MDA-MB-231 HM), HER2-type breast cancer cell lines (HCC1954, JIMT1), luminal-type breast cancer cell lines (T47D, MCF7) was detected by CCK8 colorimetric method. The experimental steps and drug concentrations are the same as the above tumor cell growth experiment. The IC 50 See Table 3:

[0209] As can be seen from the results, hetero(aromatic) ring-substituted cyclic diamines (JW02, JW04, JW05, JW07, JW10), cisplatin and verapamil can all inhibit the growth of different types of breast cancer cell lines. In some breast cancer cells, the IC 50 of hetero(aromatic) ring-substituted cyclic diamines is significantly lower than that of cisplatin or verapamil, and has a significant inhibitory effect on breast cancer cells (Table 3).

[0210] Specifically, the effect of JW02 on breast cancer MDA-MB-231, MDA-MB-453, SPC-2, MCF7 is better than that of verapamil and cisplatin;

[0211] The effect of JW04 on breast cancer MDA-MB-453 and MCF7 is better than that of verapamil and cisplatin;

[0212] The effect of JW05 on breast cancer MDA-MB-453, SPC-2, JIMT1 and MCF7 is better than that of verapamil and cisplatin;

[0213] The effect of JW07 on breast cancer MDA-MB-453, 4173, SPC-46 is better than that of verapamil and cisplatin;

[0214] The effect of JW10 on breast cancer MDA-MB-231, MDA-MB-453, SPC-2, HCC1954 and MCF7 is better than that of verapamil and cisplatin.

[0215] Table 3 IC 50

[0216] (4) The effect of JW02 on inhibiting tumor growth in immunodeficient nude mice

[0217] Cell inoculation: Take the logarithmic growth phase of MDA-MB-231 LM2 cells, adjust the cell suspension concentration to 2x10 7 Take healthy Balb / c nude mice, subcutaneously inject MDA-MB-231 LM2 cell suspension 100 μL per mouse on the right back of the mouse, and observe the growth of the tumor of the mouse. Experimental grouping and drug administration: take the tumor-bearing mice on the 7th day of inoculation, and select the mice with tumor size of about 80mm 3 The experimental animals are divided into 3 groups, respectively: vehicle (blank control group) / vilazodone (vilazodone) / JW02, 7 in each group. Drug administration starts on the 7th day after tumor inoculation. The control group is the same dose of drug solvent, and the drug administration dose is 5 mg / kg, with a frequency of 5 times / week, intraperitoneal injection, and the drug administration cycle is 3 weeks. The tumor size and mouse body weight are measured every two days during the drug administration period, and the tumor inhibition curve and body weight change curve are drawn. The mice are sacrificed by CO2 asphyxiation 48 hours after the last administration, and the main organs, tumor tissue and serum of the mice are collected, and the tumor weight is measured.

[0218] In the immunodeficient mouse subcutaneous tumor model of breast cancer, JW02 significantly inhibited the weight of the mouse tumor, and the effect was significantly better than that of the blank control group and the vilazodone administration group (see Figure 1, Figure 2; **: p<0.01; ****: p<0.0001).

[0219] (5) The effect of JW02 on inhibiting tumor growth in immune normal mice

[0220] Cell inoculation: Take the logarithmic growth phase of 4T1 cells, adjust the cell suspension concentration to 1x10 6 Take 30 4-6 week old Balb / c female mice, inject 4T1 cell suspension 100 μL per mouse under the mammary pad of the mouse, and observe the growth of the tumor of the mouse. Experimental grouping and drug administration: take the tumor-bearing mice on the 7th day of inoculation, and select the mice with tumor size of about 80mm 3 The experimental animals are divided into 3 groups, respectively: vehicle (blank control group) / vilazodone (vilazodone) / JW02, 7 in each group. Drug administration starts on the 7th day after tumor inoculation. The control group is the same dose of drug solvent, and the drug administration dose is 5 mg / kg, with a frequency of 5 times / week, intraperitoneal injection, and the drug administration cycle is 3 weeks. The tumor size and mouse body weight are measured every two days during the drug administration period, and the tumor inhibition curve and body weight change curve are drawn. The mice are sacrificed by CO2 asphyxiation 48 hours after the last administration, and the main organs, tumor tissue and serum of the mice are collected, and the tumor weight is measured.

[0221] In the immune normal mouse breast cancer tumor model, JW02 significantly inhibited the volume and weight of mouse tumor, and the effect was significantly better than that of the blank control group and the vilazodone administration group (see Figures 3 and 4; **: p < 0.01; ****: p < 0.0001).

[0222] (5) Effect of hetero(aryl) ring-substituted cyclic diamine compounds on degradation of PD-L1 protein

[0223] The MDA-MB-231 cell suspension in the logarithmic growth phase was adjusted in cell density, and was inoculated into a 6-well plate at a density of 4 x 104 cells per well; after the cells adhered, the medium containing vilazodone, JW02, JW04, JW05, JW07 (at a concentration of 10 μM, respectively), and JW10 (at a concentration of 5 μM) was replaced, respectively. After 48 h of incubation, the protein was collected, and Western-blot detection was performed (see Figure 5). 5

[0224] Compared with the group without administration (blank control group), the PD-L1 protein of the three administration groups of JW02, JW04, and JW10 was significantly reduced, and the PD-L1 protein of the two administration groups of JW05 and JW07 was reduced to a certain extent. This result shows that the hetero(aryl) ring-substituted cyclic diamine compounds can reduce the PD-L1 protein level to different degrees, and are a PD-L1 immunomodulator.

[0225] It should be noted that the above description is only an embodiment of the present application, and does not limit the patent scope of the present application, and any direct or indirect use of the content of the present application and the content of the drawings in other related technical fields is also included in the patent protection scope of the present application. Therefore, any technical solution obtained by logical analysis, reasoning or limited experiment on the basis of the existing technology according to the concept of the present application should be within the protection scope determined by the claims.​

Claims

1. A hetero(areno)cyclic substituted cyclic diamine compound, characterized by: The chemical structural formula of the compound is shown as (I): In the formula: X, Y, Z, W each independently represent a carbon or nitrogen atom, and when selected from a nitrogen atom, only one is nitrogen; R1 is hydrogen, methyl, cyano, halogen, haloalkyl, primary amide, hydroxyl, thiol, alkoxy, alkylthio, haloalkoxy, haloalkylthio, aralkyl, aryl; or a combination of any two identical or different groups; or a combination of any three identical or different groups; R2 is -CH2- or -CO-; R3 is -CH2-, -CH2-CH2-, -CH2-CH2-CH2-; R4 is -CH2-, -CH2-CH2-, -CH2-CH2-CH2-; R5 is benzofuran, benzopyridine, benzimidazole, benzoxazole, benzothiophene, benzopyrazole, quinoline, isoquinoline, indole, 2H-1,3-benzodioxolane 2H-chromen-2-one 1 -chloro-3-methoxybenzene 2,1,3-benzothiadiazole benzene ring, pyridine ring, naphthalene ring, six-membered heterocyclic ring, benzofused heterocyclic ring or fused heterocyclic ring; R6 is -CONH2, -SO2NH2, 1,1,1-trifluoropropan-2-amine hydrogen, methyl, cyano, halogen, haloalkyl, hydroxyl, thiol; or a combination of any two identical or different groups; or a combination of any three identical or different groups.

2. The hetero (aromatic) ring-substituted cyclic diamine compound of claim 1, wherein: In the above chemical formula, R6, if a functional group, can be combined with any carbon atom on the R5 ring; or R6 is absent; Halogen includes F, Cl, Br or I; Haloalkyl is monohaloalkane, dihaloalkane or polyhaloalkane; R1 is preferably CF3, OCF3 or SCF3; R3 is preferably -CH2-CH2-; R4 is preferably -CH2-CH2-; R5 is preferably benzofuran; R6is preferably -CONH2, -SO2NH2, 1-methylcyclopropane-1-amine or 1,1,1-trifluoropropan-2-amine X, Y, Z, W are preferably carbon atoms; m represents 1, 2 or 3; n represents 0, 1, 2, 3 or 4.

3. The hetero(aromatic) ring-substituted cyclic diamine compound according to claim 1 or 2, wherein The compounds include:

4. The hetero(arene) ring-substituted cyclic diamine compound according to claim 1 or 2, wherein The compound is in the form of its tautomer, mesomer, racemate, enantiomer, diastereomer or possible derivative based on its structure or mixture thereof.

5. The use of the hetero (aromatic) ring-substituted cyclic diamine compound according to claim 1, characterized by The use of the compound or a pharmaceutical composition comprising the compound or a derivative of the compound in the preparation of a drug for preventing and / or treating tumors.

6. Use according to claim 5, characterized in that, The compound is in the form of its pharmaceutically acceptable salt, ether, ester, prodrug, metabolite, solvate or crystal thereof; The pharmaceutically acceptable salt includes hydrochloride, hydrobromide, fumarate, acetate, citrate, sulfate, methanesulfonate, formate or trifluoroacetate.

7. Use according to claim 5 or 6, characterized in that, The drug is a tablet, injection, capsule, oral solution, pill, granule, powder, aerosol, patch, ointment, paint or suppository.

8. Use according to claim 5 or 6, characterized in that, The drug is a conventional anti-tumor drug; including a chemotherapy drug, a biological targeted therapy drug, a metabolic therapy drug or an immunotherapy drug.

9. Use according to claim 5 or 6, characterized in that, The compound is used in the preparation of a PD-L1 immunomodulator related drug.

10. Use according to claim 5 or 6, characterized in that, The effects of the compound include inhibiting tumor growth and / or metastasis; The compound or its composition is used as a drug for preventing and / or treating leukemia, lymphoma, breast cancer, melanoma, ovarian cancer, colon cancer, cervical cancer, lung cancer, prostate cancer, esophageal cancer, glioma, renal cancer, pancreatic cancer, nasopharyngeal carcinoma, liver cancer, gastric cancer.

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