mIDH1 / NAMPT DUAL-TARGET INHIBITOR AND USE THEREOF

Abstract

Disclosed is an mIDH1 / NAMPT dual-target inhibitor, which is selected from a compound having a structure as shown in formula (I) or formula (II), or a pharmaceutically acceptable salt, racemate, stereoisomer, prodrug, or solvent compound thereof. The dual-target inhibitor of the present invention can effectively overcome the shortcomings of a single mIDH1 inhibitor and single NAMPT inhibitor having poor efficacy against solid tumors. Said inhibitor can inhibit the malignant proliferation of tumors, has a good therapeutic effect, has low toxicity, and can penetrate the blood-brain barrier, and it is not easy to acquire drug resistance to said inhibitor. Moreover, the inhibitor can be used to prepare a medicament for treating cancer or tumor-related diseases, the cancer or tumor-related diseases comprising multiple myeloma, leukemia, breast cancer, prostate cancer, lung cancer, liver cancer, gastric cancer, bone cancer, brain cancer, head and neck cancer, intestinal cancer, pancreatic cancer, bladder cancer, testicular cancer, ovarian cancer, and endometrial cancer.

Description

mIDH1 / NAMPT dual-target inhibitors and their applications Technical Field

[0001] The present invention belongs to the field of medicinal chemistry and relates to a class of mIDH1 / NAMPT dual-target inhibitors and applications thereof. Background Art

[0002] Isocitrate dehydrogenase (IDH) is a key metabolic enzyme involved in the tricarboxylic acid cycle. The wild-type protein catalyzes the oxidative decarboxylation of isocitrate to α-ketoglutarate (α-KG), while generating CO2 and NADPH. Researchers have identified IDH1 mutations (mainly in the cytoplasm) in various cancer types. Mutations often occur at position 132 of IDH1. Mutated IDH1 (mIDH1) acquires new catalytic activity, further converting α-KG to 2-hydroxyglutarate (2-HG) while oxidizing NADPH to NADP. + . 2-HG is a carcinogenic metabolite, and its large accumulation can induce the occurrence and development of tumors. Therefore, selective inhibition of mIDH1 is a safe and effective therapeutic strategy for treating cancer. Currently, the selective mIDH1 inhibitors ivosidenib (AG-120) and olutamicinib (FT-2102) have been approved by the FDA for the treatment of acute myeloid leukemia (AML). However, the existing mIDH1 inhibitors have poor therapeutic effects on solid tumors, and resistance to mIDH1 inhibitors has emerged and is becoming increasingly serious.

[0003] Nicotinamide phosphoribosyltransferase (NAMPT) is located in the cytoplasm and is a + A key rate-limiting enzyme in the salvage synthesis pathway, highly expressed in tumors, continuously provides tumors with the necessary NAD for growth. + , which is closely related to tumor growth. Therefore, selective inhibition of NAMPT is also an effective strategy for treating cancer. However, clinical trials of NAMPT inhibitors as single drugs for the treatment of solid tumors and hematological tumors have shown that their dose-limiting toxicity leads to poor anti-tumor effects. Currently, no selective NAMPT inhibitor has been approved by the FDA for the treatment of cancer.

[0004] Based on the above research status, small molecule inhibitors that can simultaneously inhibit mIDH1 and NAMPT are needed for the treatment of various cancers.

[0005] Summary of the Invention

[0006] The purpose of the present invention is to provide a dual-target inhibitor that can penetrate the brain and effectively solve the shortcomings of single mIDH1 inhibitors and single NAMPT inhibitors, such as poor efficacy against solid tumors.

[0007] The purpose of the present invention is achieved through the following technical solutions:

[0008] As one of the technical solutions of the present invention, the mIDH1 / NAMPT dual-target inhibitor is selected from the compound having the structure shown in Formula I or its pharmaceutically acceptable salt, racemate, stereoisomer, prodrug or solvent compound:

[0009] wherein Ring A is selected from a substituted or unsubstituted 6-membered saturated or unsaturated heterocyclic ring, a benzene ring and a 5-membered saturated or unsaturated heterocyclic ring, or a 5-6-membered saturated or unsaturated heterocyclic ring and a 5-6-membered saturated or unsaturated heterocyclic ring; the substituent of the 5-6-membered saturated or unsaturated heterocyclic ring is at least one of halogen, amino, hydroxyl, carboxyl, cyano, olefin, and amide; the heteroatom in the 6-membered saturated or unsaturated heterocyclic ring, the benzene ring and a 5-membered saturated or unsaturated heterocyclic ring, or the 5-6-membered saturated or unsaturated heterocyclic ring and a 5-6-membered saturated or unsaturated heterocyclic ring is selected from an oxygen atom and a nitrogen atom;

[0010] X is selected from a linear or branched alkylene chain, a 1,4-piperazine group or a 1,3-pyrrolidino group; X may be interrupted one or more times by one, more or any combination of the following groups: -O-, -CONH-, NHCO-, -NHCONH-, -NH-, -S-, alkenylene; the hydrogen of X may be substituted by one or more of the following groups: halogen, amino, hydroxyl, carboxyl, amide, cyano, alkene, alkyne, C1-C4 alkoxy, C1-C6 alkyl;

[0011] L is selected from a covalent bond, a substituted or unsubstituted benzene ring, a 5-6 membered heterocycle, a C1-C4 alkenyl group, a C1-C4 alkoxy group, or a C1-C4 amide group; the substituent of the benzene ring is at least one of halogen, amino, hydroxyl, carboxyl, cyano, olefin, and amide;

[0012] Ring B is selected from a substituted or unsubstituted benzene ring, a 5-6 membered saturated or unsaturated heterocyclic ring; the substituent of the benzene ring is at least one of halogen, amino, hydroxyl, carboxyl, cyano, olefin, and amide; the 5-6 membered saturated or unsaturated heterocyclic ring contains 1-2 heteroatoms selected from oxygen and nitrogen atoms;

[0013] R 4 Selected from hydrogen or C1-C4 alkyl;

[0014] R 5 is selected from halogen, C1-C4 alkyl, substituted or unsubstituted phenyl, hydroxyl or tert-butoxy, and the substituent of the phenyl ring is at least one of halogen, amino, hydroxyl, carboxyl, cyano, olefin, and amide;

[0015] or R 4 With R 5 Combination to form a benzene ring.

[0016] Preferably, in the compound shown in Formula I, Ring A is selected from:

[0017] X is selected from: m=an integer of 0-8, specifically, m=0, 1, 2, 3, 4, 5, 6, 7, 8; n=an integer of 0-8, specifically, n=0, 1, 2, 3, 4, 5, 6, 7, 8; p=an integer of 0-8, specifically, p=0, 1, 2, 3, 4, 5, 6, 7, 8, preferably an integer of 1-4;

[0018] More preferably, in the compound of formula I, L is selected from a benzene ring Phenoxy Pyridine ring Pyrimidine ring or covalent bonds;

[0019] Ring B is selected from benzene ring Pyridine ring Pyrimidine ring or imidazole ring

[0020] R 4 is selected from hydrogen or methyl;

[0021] R 5 Selected from F, methyl, phenyl, hydroxy or tert-butoxy;

[0022] or R 4 With R 5 Combination to form a benzene ring.

[0023] In the compound shown in Formula I, Ring A is selected from:

[0024] X is selected from: m=0 n=0

[0025] L is selected from a benzene ring;

[0026] Ring B is selected from a benzene ring or a pyridine ring;

[0027] R 4 selected from methyl;

[0028] R 5 Selected from F, methyl, phenyl, hydroxy or tert-butoxy;

[0029] or R 4 With R 5 Combination to form a benzene ring.

[0030] As a further preferred embodiment of the present invention, the mIDH1 / NAMPT dual-target inhibitor is selected from the compounds represented by Formula Ia, Ib, Ic, or pharmaceutically acceptable salts, racemates, stereoisomers, prodrugs, or solvates thereof:

[0031] Among them, ring A, X, L, ring B, R 4 、R 5 The same as the compound shown in formula I.

[0032] As another technical solution of the present invention, the mIDH1 / NAMPT dual-target inhibitor is selected from the compound shown in Formula II or its pharmaceutically acceptable salt, racemate, stereoisomer, prodrug or solvent compound:

[0033] wherein Ring A is selected from a substituted or unsubstituted 6-membered saturated or unsaturated heterocyclic ring, a benzene ring and a 5-membered saturated or unsaturated heterocyclic ring, or a 5-6-membered saturated or unsaturated heterocyclic ring and a 5-6-membered saturated or unsaturated heterocyclic ring; the substituent of the 5-6-membered saturated or unsaturated heterocyclic ring is at least one of halogen, amino, hydroxyl, carboxyl, cyano, olefin, and amide; the heteroatom in the 6-membered saturated or unsaturated heterocyclic ring, the benzene ring and a 5-membered saturated or unsaturated heterocyclic ring, or the 5-6-membered saturated or unsaturated heterocyclic ring and a 5-6-membered saturated or unsaturated heterocyclic ring is selected from an oxygen atom and a nitrogen atom;

[0034] X is selected from a linear or branched alkylene chain, a 1,4-piperazine group or a 1,3-pyrrolidino group; X may be interrupted one or more times by one, more or any combination of the following groups: -O-, -CONH-, NHCO-, -NHCONH-, -NH-, -S-, alkenylene; the hydrogen of X may be substituted by one or more of the following groups: halogen, amino, hydroxyl, carboxyl, amide, cyano, alkene, alkyne, C1-C4 alkoxy, C1-C6 alkyl;

[0035] L is selected from a covalent bond, a substituted or unsubstituted benzene ring, a 5-6 membered heterocycle, a C1-C4 alkenyl group, a C1-C4 alkoxy group, or a C1-C4 amide group; the substituent of the benzene ring is at least one of halogen, amino, hydroxyl, carboxyl, cyano, olefin, and amide;

[0036] Ring B is selected from a substituted or unsubstituted benzene ring, a 5-6 membered saturated or unsaturated heterocyclic ring; the substituent of the benzene ring is at least one of halogen, amino, hydroxyl, carboxyl, cyano, olefin, and amide; the 5-6 membered saturated or unsaturated heterocyclic ring contains 1-2 heteroatoms selected from oxygen and nitrogen atoms;

[0037] R 1 Selected from sulfur, -NH-, -N(CH3)- or -CH-;

[0038] R 2 is selected from nitrogen or -CH-;

[0039] R 3 selected from sulfur or -CH-;

[0040] R 4 Selected from hydrogen or C1-C4 alkyl;

[0041] R 5 is selected from halogen, C1-C4 alkyl, substituted or unsubstituted phenyl, hydroxyl or tert-butoxy, and the substituent of the phenyl ring is at least one of halogen, amino, hydroxyl, carboxyl, cyano, olefin, and amide;

[0042] or R 4 With R 5 Combination to form a benzene ring.

[0043] Preferably, in the compound shown in Formula II, Ring A is selected from:

[0044] X is selected from: m=an integer from 0 to 8; n=an integer from 0 to 8; p=an integer from 0 to 8;

[0045] L is selected from a benzene ring, a phenoxy group, a pyridine ring, a pyrimidine ring or a covalent bond;

[0046] Ring B is selected from a benzene ring, a pyridine ring, a pyrimidine ring or an imidazole ring;

[0047] R 1 Selected from sulfur, -NH-, -N(CH3)- or -CH-;

[0048] R 2 is selected from nitrogen or -CH-;

[0049] R 3 selected from sulfur or -CH-;

[0050] R 4 is selected from hydrogen or methyl;

[0051] R 5 Selected from F, methyl, phenyl, hydroxy or tert-butoxy;

[0052] or R 4 With R 5 Combination to form a benzene ring.

[0053] More preferably, in the compound shown in Formula II, Ring A is selected from:

[0054] X is selected from: m=an integer from 0 to 8; n=an integer from 0 to 8; p=an integer from 0 to 8;

[0055] L is selected from a benzene ring;

[0056] Ring B is selected from a benzene ring, a pyridine ring or a pyrimidine ring;

[0057] R 1 Selected from sulfur, R 2 Selected from -CH-, R 3 selected from -CH-; or R 1 Selected from -CH-, R 2 Selected from -CH-, R 3 Selected from S; or R 1 Selected from -NH-, R 2 Selected from -CH-, R 3 selected from -CH-;

[0058] R 4 selected from methyl;

[0059] R 5 Selected from methyl.

[0060] Most preferably, in the compound of formula II, ring A is selected from:

[0061] X is selected from: n=0;

[0062] L is selected from a benzene ring;

[0063] Ring B is selected from a benzene ring, a pyridine ring or a pyrimidine ring;

[0064] R 1 Selected from -NH-, R 2 Selected from -CH-, R 3 selected from -CH-;

[0065] R 4 selected from methyl;

[0066] R 5 Selected from methyl.

[0067] As a further preferred embodiment of the present invention, the mIDH1 / NAMPT dual-target inhibitor is selected from the compound having the structure shown in Formula IIa or a pharmaceutically acceptable salt, racemate, stereoisomer, prodrug or solvate thereof:

[0068] Among them, ring A, X, L, ring B, R 1 、R 2 、R 3 、R 4 、R 5The same as the compound shown in formula II.

[0069] Specifically, the mIDH1 / NAMPT dual-target inhibitor is selected from the compounds with the following structures:

[0070] Pharmaceutically acceptable salts are acid addition salts of the compounds represented by Formula I and Formula II. The acids used to form the salts include inorganic acids and organic acids; the inorganic acids include hydrochloric acid, sulfuric acid, phosphoric acid, and methanesulfonic acid; the organic acids include acetic acid, trichloroacetic acid, propionic acid, butyric acid, maleic acid, p-toluenesulfonic acid, malic acid, malonic acid, cinnamic acid, citric acid, fumaric acid, camphoric acid, digluconic acid, aspartic acid, and tartaric acid.

[0071] Another object of the present invention is to provide a pharmaceutical composition comprising the mIDH1 / NAMPT dual-target inhibitor and a pharmaceutically acceptable carrier.

[0072] A pharmaceutically acceptable carrier refers to an excipient or diluent that does not cause significant irritation to an organism and does not interfere with the biological activity and properties of the administered compound.

[0073] The compounds represented by Formula I and Formula II of the present invention have dual mIDH1 / NAMPT inhibitory activity, can effectively overcome the shortcomings of single mIDH1 inhibitors and single NAMPT inhibitors in terms of poor efficacy against solid tumors, can inhibit the malignant proliferation of cancer, have good therapeutic effects, low toxicity, and are not prone to drug resistance. In addition, the compounds of the present invention can penetrate the blood-brain barrier. Therefore, the compounds of the present invention can be prepared into dual mIDH1 / NAMPT inhibitors and can be used to prepare drugs for treating cancer or tumor-related diseases.

[0074] Another object of the present invention is to provide the use of the mIDH1 / NAMPT dual-target inhibitor in the preparation of drugs for diseases mediated by mIDH1 and / or NAMPT.

[0075] The disease mediated by mIDH1 and / or NAMPT is cancer or tumor-related disease.

[0076] Another object of the present invention is to provide the use of the mIDH1 / NAMPT dual-target inhibitor in the preparation of drugs for treating cancer or tumor-related diseases.

[0077] Cancer or tumor-related diseases include, but are not limited to, multiple myeloma, leukemia, breast cancer, prostate cancer, lung cancer, liver cancer, stomach cancer, bone cancer, brain cancer, head and neck cancer, intestinal cancer, pancreatic cancer, bladder cancer, testicular cancer, ovarian cancer, and endometrial cancer.

[0078] Unless otherwise specified, the terms used in the present invention generally have the following meanings.

[0079] Me is methyl, Et is ethyl, and Boc is tert-butyloxycarbonyl.

[0080] The term "alkyl" refers to a straight or branched chain saturated hydrocarbon group having the stated number of carbon atoms.

[0081] The term "C1-C4 alkyl" refers to a straight or branched chain saturated hydrocarbon group having 1 to 4 carbon atoms. C1-C4 alkyl includes, but is not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, and tert-butyl.

[0082] The term "alkoxy" refers to an O-alkyl group. The term "C1-C4 alkoxy" refers to a group having an O-C1-C4 alkyl group.

[0083] The term "halogen" refers to fluorine, chlorine, bromine or iodine, preferably fluorine, chlorine or bromine. BRIEF DESCRIPTION OF THE DRAWINGS

[0084] Figure 1 shows H&E stained pathological sections of the heart, liver, spleen, lung, and kidney tissues of ICR mice 7 days after administration.

[0085] FIG2 is a curve showing the body weight changes of nude mice with orthotopic transplanted tumors after drug administration.

[0086] Figure 3 shows the fluorescence imaging results of the brain tissue of nude mice with orthotopic transplanted tumors after drug administration.

[0087] FIG4 shows the statistics of fluorescence intensity of brain tissue of nude mice with orthotopic transplanted tumors after drug administration.

[0088] FIG5 is a survival curve of nude mice with orthotopic transplanted tumors after drug administration.

[0089] FIG6 is a pathological section of the brain tissue of nude mice with orthotopic transplanted tumor after drug administration. DETAILED DESCRIPTION

[0090] The following examples are provided to better understand the technical solutions of the present invention, but are not intended to limit the present invention.

[0091] The experimental methods in the examples are conventional methods unless otherwise specified.

[0092] Unless otherwise specified, the test materials used in the examples are conventional biochemical reagents purchased from stores.

[0093] Example 1

[0094] Synthesis of compound 1a:

[0095] 2,4-Dichloropyrimidine (8.00 g, 53.70 mmol) and (S)-4-isopropyl-oxazolidin-2-one (6.93 g, 53.70 mmol) were dissolved in 40 mL of anhydrous DMF. Under stirring in an ice bath, a suspension of 60% NaH-mineral oil (2.60 g, 64.45 mmol) in anhydrous DMF (20 mL) was slowly added dropwise. After the addition was complete, stirring was continued under ice bath for about 1 hour. The ice bath was removed and the reaction was continued for 1 hour. The reaction was monitored by TLC to complete. 200 mL of ice water was slowly added to the reaction solution, accompanied by the precipitation of a large amount of white solid. 100 mL of ethyl acetate was added to dissolve the solid. The mixed solution was extracted with ethyl acetate three times, 200 mL each time. The organic phases were combined and washed with water twice (150 mL each) and saturated sodium chloride aqueous solution twice (150 mL each). The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was separated by flash column chromatography (PE:EA=15:1, v:v) to obtain 8.16 g of compound 1a as a white solid product in a yield of 62.6%. 1 H NMR(500MHz, CDCl3-d)δ8.52(d,J=5.8Hz,1H),8.23(d,J=5.8Hz,1H),4.94-4.77(m,1H ), 4.50-4.26 (m, 2H), 2.72-2.55 (m, 1H), 1.04 (d, J = 7.0Hz, 3H), 0.93 (d, J = 6.7Hz, 3H).

[0096] Synthesis of compound 2a:

[0097] Compound 1a (300 mg, 1.24 mmol), (S)-1-(4-nitrophenyl)-ethan-1-amine (247.5 mg, 1.49 mmol), Pd2(dba)3 (22.76 mg, 0.025 mmol), BINAP (77.37, 0.124 mmol), and potassium carbonate (346 mg, 2.48 mmol) were dissolved in 20 mL of toluene. The mixture was heated to 90°C under nitrogen and refluxed for 16 h. After completion of the reaction, 50 mL of water was added to the reaction solution, and the mixture was stirred at room temperature for 10 min. The mixture was extracted with ethyl acetate three times (50 mL each), and the organic phases were combined. The organic phases were washed with water, saturated sodium bicarbonate, and saturated brine (30 mL each), dried over anhydrous sodium sulfate, and isolated by flash column chromatography (PE:EA = 2:1, v:v) to afford 247 mg of compound 2a as a light yellow solid in a yield of 53.6%. 1H NMR (300MHz, DMSO-d6) δ8.14(d,J=5.6Hz,1H),7.58(s,1H),7.18(d,J=5.6Hz,1H),6.99(d,J=8.0Hz,2H),6.52–6.43(m, 2H),4.86(m,1H),4.67(dt,J=7.5,3.5Hz,1H),4.44–4.27(m,2H),2.08(s,1H),1.36(d,J=7.0Hz,3H),0.86–0.55(m,6H).

[0098] Synthesis of compound 3a:

[0099] Ammonium chloride (216 mg, 4.04 mmol) and reduced iron powder (226 mg, 4.04 mmol) were weighed into a 50 mL three-necked flask, and 10 mL of acetone and 5 mL of water were added. The mixture was heated to 80°C under nitrogen protection and refluxed for 0.5 h. 5 mL of acetone solution of compound 2a (300 mg, 0.81 mmol) was slowly injected into the mixed solution. After the addition was complete, stirring was continued at 80°C for 3 h. After the reaction was completed, the mixture was monitored by TLC, cooled, filtered through celite, and the filtrate was concentrated under reduced pressure. The concentrated material was dissolved in approximately 100 mL of ethyl acetate, washed three times with water and saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain compound 3a as an orange-yellow solid, which was used directly in the next step.

[0100] Synthesis of compound 4a:

[0101] Compound 3a (200 mg, 0.586 mmol), Boc-alanine (133 mg, 0.703 mmol), and DIPEA (0.194 mL, 1.17 mmol) were dissolved in 3 mL of anhydrous DMF and stirred on an ice bath for 10 min. HATU (334 mg, 0.879 mmol) was then slowly added. After the addition, the mixture was returned to room temperature and stirred for 4 h. The reaction was monitored by TLC until the end point. With stirring, the reaction solution was poured into 50 mL of ice water and extracted three times with ethyl acetate. The organic phases were combined and washed sequentially with water, saturated sodium bicarbonate, and saturated brine (20 mL each), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and separated by flash column chromatography (PE:EA=2:1, v:v) to give 256 mg of compound 4a as a light yellow solid in a yield of 85.3%. 1H NMR(300MHz, CDCl3)δ8.16(d,J=5.9Hz,1H),7.83(s,1H),7.53–7.46(m,3H),7.26(s,1H),5.20(s,1H),5.01(s,1H),4.68–4.54(m,1H),4 .37–4.16(m,2H),3.51(q,J=6.1Hz,2H),2.62(t,J=6.0Hz,2H),2.48–2.00(m,1H),1.54(d,J=6.9Hz,3H),1.45(s,9H),0.84–0.60(m,6H).

[0102] Synthesis of compound I-8:

[0103] Compound 4a (300 mg, 0.585 mmol) was placed in a 50 mL eggplant-shaped bottle and slowly added dropwise 3 mL of After the addition of 4M hydrochloric acid EA, stirring was continued on ice for about 5 hours, and the reaction was monitored by TLC until the end point. Filtration was performed to obtain 195 mg of a white solid powder of the hydrochloride salt. Trans-3-(3-pyridyl)acrylic acid (78 mg, 0.522 mmol), the hydrochloride solid powder, and DIPEA (0.43 ml, 2.61 mmol) were dissolved in 3 mL of anhydrous DMF and stirred on ice for 15 minutes. HATU (248 mg, 0.653 mmol) was then slowly added, and the mixture was returned to room temperature and stirred for 4 hours. The reaction was monitored by TLC for completion. The reaction solution was poured into 30 mL of ice water and extracted three times with ethyl acetate. The organic phases were combined and washed successively with water, saturated sodium bicarbonate, and saturated brine (30 mL each time), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and separated by flash column chromatography (DCM:MeOH=40:1, v:v) to obtain 98 mg of compound I-8 as a light yellow solid in a yield of 41.5%. 1 H NMR(300MHz, CDCl3)δ8.64(d,J=38.6Hz,2H),8.23–8.04(m,2H),7.78(d,J=7.9 Hz,1H),7.63–7.44(m,4H),7.26(s,1H),6.89(s,1H),6.51(d,J=15.7Hz,1H),5. 02–5.00(m,1H),4.63–4.60(m,1H),4.33–4.21(m,2H),3.77–3.70(m,2H),2.73 (m,2H),2.48–2.34(m,1H),1.46(d,J=9.0Hz,3H),0.70(dd,J=24.0,6.0Hz,6H). 13CNMR(75MHz,DMSO-d6)δ170.51,164.47,161.00,158.75,156.69,154.36,149.98,148.97,140.47,137.48,135.13,13 3.82,130.67,125.74,124.21,123.87,118.98,97.44,62.72,57.86,49.56,40.08,38.34,33.77,26.89,17.69,13.73. ESI-HRMS m / z:544.2666[M+H] + .

[0104] Example 2

[0105] Synthesis of compound 5a:

[0106] The synthesis method of compound 5a was based on the synthesis of compound 4a, except that Boc-alanine was replaced with Boc-glycine. Compound 3a (200 mg, 0.586 mmol) and Boc-glycine (123 mg, 0.703 mmol) were added to obtain 249 mg of compound 5a as a light yellow solid with a yield of 85.3%. 1 H NMR(300MHz,DMSO-d6)δ9.75(s,1H),8.16(d,J=5.6Hz,1H),7.70–7.55(m,1H),7.47( d,J=8.3Hz,2H),7.31–7.23(m,2H),7.20(d,J=5.6Hz,1H),6.95(d,J=7.7Hz,1H),4.9 7(s,1H),4.64(dq,J=8.3,4.3,3.6Hz,1H),4.33(tdd,J=12.4,8.8,3.2Hz,2H),3.70( d,J=6.1Hz,2H),2.04(m,1H),1.42(d,J=6.9Hz,3H),1.40(s,9H),0.80–0.50(m,6H).

[0107] Synthesis of compound I-10:

[0108] The synthesis method of compound I-10 refers to the synthesis of compound I-8, except that compound 4a is replaced by compound 5a, and compound 5a (200 mg, 0.401 mmol) and trans-3-(3-pyridyl)acrylic acid (62 mg, 0.415 mmol) are added to obtain 91 mg of compound I-10 as a white solid with a yield of 52.5%. 1H NMR (300MHz, DMSO-d6) δ9.97 (s, 1H), 8.79 (d, J = 2.2Hz, 1H), 8.58 (dd, J = 4.7, 1.6Hz,1H),8.44(t,J=5.8Hz,1H),8.17(d,J=5.8Hz,1H),8.03(dt,J=8.0,1.9Hz,1H) ,7.77(s,1H),7.50(ddd,J=12.8,6.7,4.3Hz,4H),7.25(dd,J=12.3,7.0Hz,3H),6.91( d,J=15.9Hz,1H),4.99(s,2H),4.64(dt,J=7.8,3.4Hz,1H),4.42–4.25(m,2H),4.04(d ,J=5.8Hz,2H),1.97(d,J=15.3Hz,1H),1.43(d,J=7.0Hz,3H),0.67(d,J=56.5Hz,6H). 13 C NMR(75MHz,DMSO-d6)δ167.31,164.82,160.38,157.86,156.98,154.30,149.78,148.78,140.56,137.10,135.54,134 .24,130.75,125.82,124.03,123.99,119.09,97.41,62.79,57.93,49.65,42.76,28.94,26.91,23.19,17.68,13.73. ESI-HRMS m / z:530.2508[M+H] + .

[0109] Example 3

[0110] Synthesis of compound 6a:

[0111] The synthesis method of compound 6a refers to the synthesis of compound 4a, except that Boc-alanine is replaced by Boc-butyric acid. Compound 3a (200 mg, 0.586 mmol) and Boc-butyric acid (143 mg, 0.703 mmol) are added to obtain 273 mg of compound 6a as a light yellow solid with a yield of 88.6%. 1H NMR (300MHz, CDCl3) δ8.83(s,1H),8.15(d,J=5.8Hz,1H),7.56(d,J=8.1Hz,2H),7.45(d, J=5.8Hz,1H),7.26(s,1H),6.06–5.78(m,1H),4.83(t,J=6.5Hz,1H),4.62(dt,J=7.6,3. 4Hz,1H),4.35–4.19(m,2H),3.26(d,J=6.2Hz,2H),2.43–2.36(m,2H),2.07(s,1H),1.88 (dd, J = 7.5, 4.8 Hz, 2H), 1.54 ( d, J = 6.9 Hz, 3H), 1.47 ( s, 9H), 0.72 ( dd, J = 19.9, 7.0 Hz, 6H).

[0112] Synthesis of compound I-11:

[0113] The synthesis method of compound I-11 refers to the synthesis of compound I-8, except that compound 4a is replaced by compound 6a, and compound 6a (200 mg, 0.380 mmol) and trans-3-(3-pyridyl)acrylic acid (58 mg, 0.390 mmol) are added to obtain 89 mg of compound I-11 as a white solid with a yield of 49.2%. 1 H NMR (300MHz, DMSO-d6) δ9.84(s,1H),8.75(s,1H),8.55(d,J=4.8Hz,1H),8.19(d,J=22.3Hz ,2H),7.97(d,J=8.0Hz,1H),7.73(s,1H),7.46(t,J=9.1Hz,4H),7.21(dd,J=11.7,6.6Hz,3H ),6.72(d,J=15.9Hz,1H),4.93(s,1H),4.62(s,1H),4.33(d,J=12.4Hz,2H),3.28–3.11(m,2 H), 2.33 (t, J = 7.5Hz, 2H), 1.97–1.67 (m, 3H), 1.40 (d, J = 7.0Hz, 3H), 0.79 (d, J = 42.7Hz, 6H). 13C NMR(75MHz,DMSO-d6)δ170.51,164.47,161.00,158.75,156.71,149.98,148.97,140.47,137.48,135.13,133.82,13 0.67,125.74,124.21,123.87,118.98,97.44,62.72,57.86,49.60,38.34,33.77,26.89,25.19,23.27,17.69,13.72. ESI-HRMS m / z:558.2827[M+H] + .

[0114] Example 4

[0115] Synthesis of compound 7a:

[0116] The synthesis method of compound 7a was the same as that of compound 4a, except that Boc-alanine was replaced by Boc-valeric acid. Compound 3a (200 mg, 0.586 mmol) and Boc-valeric acid (153 mg, 0.703 mmol) were added to obtain 283 mg of compound 7a as a light yellow solid in a yield of 89.2%. 1 HNMR (300MHz, DMSO-d6) δ9.72 (s, 1H), 8.16 (d, J = 5.6Hz, 1H), 7.60 (s, 1H), 7.50–7. 39(m,2H),7.30–7.13(m,3H),6.72(s,1H),4.96(s,1H),4.68–4.58(m,1H),4.40–4 .25(m,2H),2.98–2.87(m,2H),2.27(t,J=7.3Hz,2H),2.09(s,1H),1.61–1.49(m,2 H), 1.41 (d, J = 7.0Hz, 3H), 1.37 (s, 9H), 1.28–1.23 (m, 2H), 0.66 (d, J = 56.7Hz, 6H).

[0117] Synthesis of compound I-12:

[0118] The synthesis method of compound I-12 is the same as that of compound I-8, except that compound 4a is replaced by compound 7a. Compound 7a (200 mg, 0.370 mmol) and trans-3(3-pyridyl)acrylic acid (56 mg, 0.378 mmol) are added to obtain 73 mg of compound I-12 as a white solid with a yield of 46.3%. 1H NMR (300MHz, DMSO-d6) δ9.82(d,J=6.8Hz,1H),8.76(s,1H),8.56(d,J=4.8Hz,1H),8.19(q,J=7 .0,6.3Hz,2H),8.00(d,J=8.0Hz,1H),7.85(d,J=22.3Hz,1H),7.58–7.38(m,4H),7.34–7.08(m, 3H),6.84–6.63(m,1H),4.95(s,1H),4.62(s,1H),4.34(d,J=10.7Hz,2H),3.21(q,J=6.4Hz,2H ), 2.31 (t, J = 7.4Hz, 2H), 2.29-2.09 (m, 1H), 1.64-1.49 (m, 4H), 1.24 (s, 3H), 0.73-0.50 (m, 6H). 13 C NMR(75MHz,DMSO-d6)δ198.55,197.56,172.58,164.01,161.01,158.73,156.70,154.35,149.98,149.40,140.59,137.86,137.48,134.3 6,130.92,125.72,123.67,120.57,119.04,97.41,62.71,57.85,49.60,44.59,42.59,41.23,29.22,28.21,26.88,23.23,17.68,13.72. ESI-HRMS m / z:572.2976[M+H] + .

[0119] Example 5

[0120] Synthesis of compound 8a:

[0121] The synthesis method of compound 8a was the same as that of compound 4a, except that Boc-alanine was replaced with 1-Boc-4-piperidinic acid. Compound 3a (200 mg, 0.586 mmol) and 1-Boc-4-piperidinic acid (161 mg, 0.703 mmol) were added to obtain 289 mg of compound 8a as a light yellow solid in a yield of 89.1%. 1H NMR(300MHz, CDCl3)δ8.15(d,J=5.8Hz,1H),7.50–7.43(m,3H),7.36(s,1H), 7.26(s,1H),5.71(s,1H),5.09–4.94(m,1H),4.61(dd,J=8.0,3.9Hz,1H),4.4 0–4.05(m,6H),2.48–2.30(m,1H),1.90(d,J=13.0Hz,2H),1.74(qd,J=12.3, 4.5Hz, 2H), 1.53 (d, J = 7.1Hz, 3H), 1.48 (s, 9H), 0.72 (dd, J = 19.1, 6.9Hz, 6H).

[0122] Synthesis of compound I-13:

[0123] The synthesis method of compound I-13 is the same as that of compound I-8, except that compound 4a is replaced by compound 8a. Compound 8a (200 mg, 0.362 mmol) and trans-3-(3-pyridyl)acrylic acid (55 mg, 0.369 mmol) are added to obtain 75 mg of compound I-13 as a white solid with a yield of 47.6%. 1 H NMR(300MHz,DMSO-d6)δ9.80(s,1H),8.88(s,1H),8.66–8.46(m,1H),8.17(t, J=6.8Hz,2H),7.76–7.31(m,6H),7.22(dd,J=13.9,6.9Hz,3H),4.96(s,1H),4 .63(d,J=7.6Hz,1H),4.54–4.19(m,4H),3.30(s,2H),3.18(s,1H),2.82–2.44 (m, 3H), 1.86 (d, J = 13.1Hz, 2H), 1.42 (d, J = 7.0Hz, 3H), 0.66 (d, J = 56.0Hz, 6H). 13 C NMR(75MHz,DMSO-d6)δ170.85,164.33,160.23,157.64,157.03,154.30,149.57,148.58,140.13,137.58,134.88,134.23,130.86,1 25.68,124.47,124.02,118.95,97.42,62.78,57.93,49.67,38.42,35.91,31.47,29.38,28.93,28.66,26.89,22.67,17.67,13.71. ESI-HRMS m / z:584.2983[M+H] + .

[0124] Example 6

[0125] Synthesis of compound 9a:

[0126] The synthesis method of compound 9a was based on the synthesis of compound 4a, except that Boc-alanine was replaced with 1-Boc-pyrrolidine-3-carboxylic acid. Compound 3a (200 mg, 0.586 mmol) and 1-Boc-pyrrolidine-3-carboxylic acid (151 mg, 0.703 mmol) were added to obtain 273 mg of light yellow solid 9a with a yield of 86.7%. 1 H NMR(300MHz, DMSO-d6)δ7.92(d,J=5.8Hz,1H),7.32–7.12(m,3H),7.04(s,1H),7.02(s,1H),4.77(s,1H),4.44–4.29(m,1H),4.14–3.79(m, 2H),3.59–3.00(m,4H),2.75(d,J=10.1Hz,1H),1.94(m,2H),1.68(s,1H),1.29(d,J=6.9Hz,3H),1.23(s,9H).0.83(dd,J=17.3,5.6Hz,6H).

[0127] Synthesis of compound I-14:

[0128] The synthesis method of compound I-14 refers to the synthesis of compound I-8, except that compound 4a is replaced by compound 9a, and compound 9a (200 mg, 0.371 mmol) and trans-3-(3-pyridyl)acrylic acid (57 mg, 0.380 mmol) are added to obtain 74 mg of compound I-14 as a white solid with a yield of 47.2%. 1H NMR(500MHz,DMSO-d6)δ10.06(d,J=18.0Hz,1H),8.90(s,1H),8.57(td,J=4.8,1.6Hz,1H),8.22–8.18(m,2H) ,7.79(s,1H),7.55–7.50(m,3H),7.46(td,J=7.4,4.7Hz,1H),7.27(d,J=8.0Hz,2H),7.22(d,J=6.0Hz,1H),7 .18(s,1H),4.96(s,1H),4.64(s,1H),4.38–4.32(m,2H),4.01–3.98(m,1H),3.90–3.81(m,1H),3.78–3.70(m ,1H),3.66–3.55(m,1H),3.29–3.16(m,1H),2.28–2.14(m,2H),1.43(d,J=7.0Hz,3H),0.63(d,J=6.0Hz,6H). 13 C NMR(75MHz,DMSO-d6)δ170.79,170.40,163.06,160.86,158.54,156.76,15 4.34,149.97,149.33,140.77,137.23,136.93,134.50,130.82,125.78,123 .75,121.90,121.78,119.15,97.43,62.73,57.87,49.60,48.65,48.46,45 .76,45.39,44.56,42.66,39.50,29.47,28.08,26.90,23.24,17.68,13.73. ESI-HRMS m / z:570.2818[M+H] + .

[0129] Example 7

[0130] Synthesis of compound 14a:

[0131] (S)-(-)-1-(4-Bromophenyl)ethylamine (4 g, 20 mmol), 4-nitrophenylboronic acid (3.68 g, 22 mmol), tetrakistriphenylphosphine palladium (1.1 g, 1 mmol), and cesium carbonate (13.04 g, 40 mmol) were dissolved in a mixed solvent of 20 mL of water and 100 mL of 1,4-dioxane, heated to 90°C under nitrogen for 4 h, and the reaction was monitored by TLC. After the reaction, the reaction solution was cooled to room temperature and concentrated under reduced pressure to remove 1,4-dioxane. 150 mL of ethyl acetate and 100 mL of water were added to dissolve the diluted concentrate and transferred to a separatory funnel. The solution was extracted three times with 200 mL of ethyl acetate each time, and the organic phases were combined. The organic phases were washed with 2 mol / L aqueous sodium hydroxide solution, water, and saturated brine once each, 150 mL each time. The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain 5 g of crude compound 14a as a yellow solid, which was used directly in the next reaction without further purification. 1 H NMR (400MHz, CDCl3) δ8.29(d,J=8.8Hz,2H),7.73(d,J=8.8Hz,2H),7.60(d,J=8 .3Hz, 2H), 7.49 (d, J = 8.2Hz, 2H), 4.21 (q, J = 6.6Hz, 1H), 1.43 (d, J = 6.6Hz, 3H).

[0132] Synthesis of compound 15a:

[0133] Compound 14a (1.50 g, 6.19 mmol), compound 1a (1.5 g, 6.19 mmol) and DIPEA (4 g, 31 mmol) were dissolved in 5 mL of DMSO and heated to 110°C under nitrogen for 16 h. The reaction was monitored by TLC. After the reaction, the reaction solution was cooled to room temperature, 50 mL of ethyl acetate and 50 mL of water were added, and the mixture was stirred at room temperature for 10 min. The mixture was extracted with ethyl acetate twice, each with 50 mL. The organic phases were combined and washed successively with water and saturated sodium chloride aqueous solution, each with 50 mL each, and dried over anhydrous sodium sulfate. Finally, the mixture was purified by flash column chromatography (PE:EA=5:1, v:v) to obtain 1.14 g of compound 15a as a yellow solid in a yield of 41.2%. 1H NMR (500MHz, DMSO-d6) δ8.31(d,J=8.6Hz,2H),8.21(s,1H),7.94(d,J=8.6Hz,2H),7.74(d,J=8.1Hz,2H),7.63(d,J=7.0Hz,2H),7.59(d,J=2.8 Hz,1H),7.23(d,J=5.6Hz,1H),5.04(s,1H),4.60(s,1H),4.32(d,J=29. 0Hz, 2H), 1.72 (s, 1H), 1.49 (d, J = 7.0Hz, 3H), 0.65 (s, 3H), 0.45 (s, 3H).

[0134] Synthesis of compound 16a:

[0135] The synthesis steps of compound 16a were based on the synthesis of compound 3a. Compound 15a (600 mg, 1.34 mmol), reduced iron powder (374 mg, 6.70 mmol) and ammonium chloride (358 mg, 6.70 mmol) were added to obtain 512 mg of compound 16a as a yellow solid with a yield of 91.3%.

[0136] Synthesis of compound I-16:

[0137] The synthesis steps of compound I-16 refer to the synthesis of compound 4a, and compound 16a (150 mg, 0.36 mmol) and imidazo[1,2-a]pyridine-6-carboxylic acid (88 mg, 0.54 mmol) are added to obtain 68 mg of compound I-16 as a white solid with a yield of 33.6%. 1 H NMR (400MHz, DMSO-d6) δ10.56(s,1H),9.39(s,1H),8.33–7.71(m,8H),7.60(d,J=27.3Hz,4H),7.36(s,2H),7.18(s,1H),4. 97(s,1H),4.58(s,1H),4.28(d,J=22.9Hz,2H),1.74(s,1H),1.43(s,3H),0.63(d,J=2.48MHz,3H),0.43(d,J=1.68MHz,3H). 13C NMR (100MHz, DMSO-d6) δ163.08,161.38,159.07,157.37,154.95,151.69,147.91,145.53,142.78,138.48,136.32,130. 33,129.85,128.16,127.31,126.73,123.06,121.32,115.96,114.62,63.32,58.48,55.45,27.41,23.89,18.35,14.24. ESI-HRMS m / z:562.2566[M+H] + .

[0138] Example 8

[0139] Synthesis of compound I-17:

[0140] The synthesis method of compound I-17 refers to the synthesis of compound 4a, and compound 16a (200 mg, 0.48 mmol) and trans-3-(3-pyridyl)acrylic acid (108 mg, 0.72 mmol) are added to obtain 112 mg of compound I-17 as a light yellow solid powder with a yield of 42.6%. 1 H NMR (300MHz, DMSO-d6) δ10.39(s,1H),8.84(s,1H),8.59(d,J=3.7Hz,1H),8.18(d,J=4.7Hz,1H),8.06(d ,J=7.9Hz,1H),7.85(d,J=8.4Hz,1H),7.79(d,J=8.4Hz,2H),7.68(s,1H),7.60(dd,J=13.9,8.3Hz,4H),7 .49(dd,J=7.8,4.9Hz,1H),7.39(d,J=6.5Hz,2H),7.21(d,J=5.6Hz,1H),6.96(d,J=15.8Hz,1H),5.01(s ,1H),4.62(s,1H),4.33(d,J=10.1Hz,2H),1.77(s,1H),1.47(d,J=6.9Hz,3H),0.67(s,3H),0.46(s,3H). 13CNMR(150MHz,DMSO-d6)δ175.00,163.56,161.63,159.48,157.18,154.89,150.88,149.77,145.43,138.85, 138.29,137.38,135.71,134.60,131.01,127.34,126.60,124.64,124.53,120.15,97.85,79.02,63.20,58.37,50.39,29.51,18.29. ESI-HRMS m / z:549.2609[M+H] + .

[0141] Example 9

[0142] Synthesis of compound I-18:

[0143] The synthesis method of compound I-18 refers to the synthesis of compound 4a, and compound 16a (150 mg, 0.36 mmol) and 3-pyridyloxyacetic acid (83 mg, 0.54 mmol) are added to obtain 81 mg of compound I-18 as a white solid with a yield of 40.9%. 1 H NMR(400MHz,DMSO-d6)δ10.18(s,1H),8.44(s,1H),8.25–8.02(d,2H),7.80 (s,1H),7.66(d,J=8.3Hz,2H),7.54(dd,J=15.5,8.2Hz,4H),7.39–7.31(m,4 H),7.16(d,J=5.3Hz,1H),4.96(s,1H),4.78(s,2H),4.56(s,1H),4.26(d,J =22.0Hz,2H),1.67(s,1H),1.42(d,J=6.7Hz,3H),0.61(s,3H),0.41(s,3H). 13 C NMR(100MHz,DMSO-d6)δ166.66,161.65,155.64,154.96,154.73,153.05,142.80,138.55,138.27,138.08,136.00 ,127.31,126.68,124.69,121.83,120.62,67.64,63.29,58.44,54.13,27.41,23.96,18.61,18.34,17.26,14.17. ESI-HRMS m / z:553.2561[M+H] + .

[0144] Example 10

[0145] Synthesis of compound I-19:

[0146] The synthesis method of compound I-19 refers to the synthesis of compound 4a, and compound 16a (150 mg, 0.36 mmol) and 7-azaindole-5-carboxylic acid (88 mg, 0.54 mmol) are added to obtain 79 mg of compound I-19 as a light yellow solid with a yield of 39.1%. 1 H NMR(400MHz,DMSO-d6)δ11.96(s,1H),10.33(s,1H),8.81(d,J=1.9Hz,1H),8.57–8.5 3(m,1H),8.19–8.08(m,1H),7.85(d,J=8.6Hz,2H),7.57(dd,J=16.1,8.5Hz,6H),7.35 (d,J=4.9Hz,2H),7.17(d,J=5.5Hz,1H),6.66–6.50(m,1H),4.97(s,1H),4.57(s,1H), 4.28(d,J=19.9Hz,2H),1.77(s,1H),1.43(d,J=6.9Hz,3H),0.63(s,3H),0.43(s,3H). 13 C NMR(100MHz,DMSO-d6)δ165.84,154.99,150.20,143.39,139.21,138.41,136.78,135.67,128.60,128.34,127.14,12 6.65,124.70,123.18,121.12,119.21,101.55,97.99,63.29,58.45,46.03,29.57,27.41,23.97,18.35,14.24,9.14. ESI-HRMS m / z:562.2561[M+H] + .

[0147] Example 11

[0148] Synthesis of compound I-20:

[0149] The synthesis method of compound I-20 refers to the synthesis of compound 4a, and compound 16a (150 mg, 0.36 mmol) and 1H-pyrazolo[3,4-b]pyrimidine-5-carboxylic acid (88 mg, 0.54 mmol) are added to obtain 55 mg of compound I-20 as a light yellow solid with a yield of 27.3%. 1H NMR (500MHz, CDCl3) δ9.36 (s, 1H), 8.69 (s, 1H), 8.44 (d, J = 21.75Hz, 1H), 8.34 (d, J =21.2Hz,1H),8.26(s,1H),7.73(d,J=14.45Hz,2H),7.61(d,J=9.7Hz,2H),7.56–7. 50(m,4H),7.46(m,2H),7.24(s,1H),6.98(d,J=9.4Hz,1H),5.07(s,1H),4.57(s,1H ), 4.20-4.29 (m, 2H), 1.87 (s, 1H), 1.58 (d, J = 9.5Hz, 3H), 0.85 (s, 3H), 0.62 (s, 3H). 13 C NMR (100MHz, DMSO-d6) δ161.07,158.84,157.51,157.38,154.82,144.84,144.21,143.29,139.03,138.75,138.14,133. 09,132.24,128.67,126.37,125.49,123.55,122.99,109.64,106.39,99.56,63.01,58.59,50.78,31.59,27.45,18.29. ESI-HRMS m / z:562.2495[M+H] + .

[0150] Example 12

[0151] Synthesis of compound I-21:

[0152] Dissolve N,N'-carbonyldiimidazole (350 mg, 2.16 mmol) and triethylamine (200 μL, 1.44 mmol) in 4 mL of anhydrous tetrahydrofuran. Slowly add a solution of compound 16a (300 mg, 0.72 mmol) in anhydrous tetrahydrofuran dropwise under nitrogen protection and an ice bath. Stir for 2 h. Monitor by TLC until the reaction of compound 16a is complete. Add a solution of 3-aminomethylpyridine (94 mg, 0.86 mmol) in tetrahydrofuran dropwise. Heat to 60°C under nitrogen protection and react for 5 h. Cool the reaction solution to room temperature and concentrate under reduced pressure to remove the solvent, yielding a yellow oily liquid. The yellow oily liquid was dissolved in 100 mL of ethyl acetate, washed once with 100 mL of water and 100 mL of saturated sodium chloride aqueous solution, and dried over anhydrous sodium sulfate; flash column chromatography (DCM:CH3OH=50:1, v:v) gave 53 mg of compound I-21 as a yellow solid, with a yield of 13.4%. 1H NMR(400MHz,DMSO-d6)δ8.13(s,1H),7.76(s,1H),7.64–7.54(m,3H),7.52(dd, J=6.6,2.5Hz,2H),7.38(d,J=8.1Hz,2H),7.25(d,J=8.3Hz,4H),7.15(d,J=5.6 Hz,2H),6.58(d,J=8.4Hz,2H),5.12(s,2H),5.02–4.87(m,1H),4.57(s,1H),4. 37–4.17(m,2H),1.76(s,1H),1.40(d,J=6.9Hz,3H),0.63(s,3H),0.42(s,3H). 13 CNMR(100MHz,DMSO-d6)δ161.69,157.30,154.97,148.69,143.88,139.44,133.78,132.77,132.57,132.07,131.98,129 .35,129.24,128.07,127.55,126.49,126.39,125.74,114.73,97.88,63.29,58.45,50.35,27.40,23.97,18.33,14.23. ESI-HRMS m / z:552.2692[M+H] + .

[0153] Example 13

[0154] Route 1:

[0155] Synthesis of compound 19a:

[0156] 5-Bromo-2-pyridinecarboxaldehyde (4 g, 21.4 mmol) and (S)-(-)-tert-butylsulfenamide (3.9 g, 28.8 mmol) were dissolved in 180 mL of 1,2-dichloroethane, and cesium carbonate (21 g, 64.6 mmol) was added. The temperature was raised to 80°C under nitrogen for 5 h. After the reaction was completed as monitored by TLC, heating was stopped, and 60 mL of saturated ammonium chloride solution was added dropwise to the reaction solution. After stirring for 10 minutes, the reaction solution was transferred to a separatory funnel, and 400 mL of dichloromethane, 240 mL of saturated ammonium chloride solution, and 100 mL of water were added. The solution was shaken thoroughly and separated. The organic phase was collected and concentrated under reduced pressure to remove the solvent. 5.6 g of compound 19a was isolated by flash column chromatography (PE:EA=8:1, v:v) as a white solid in a 90% yield. 1H NMR (400MHz, CDCl3) δ8.82 (dd, J=2.3, 0.9Hz, 1H), 8.67 (s, 1H), 7.95 (qd, J=8.4, 1.5Hz, 2H), 1.30 (s, 9H).

[0157] Synthesis of compound 20a:

[0158] Compound 19a (5.6 g, 19.36 mmol) was dissolved in 230 mL of dichloromethane, and the temperature was lowered to -78°C after nitrogen substitution. 50 mL of a 1 mol / L solution of methylmagnesium bromide in tetrahydrofuran was slowly added dropwise. After completion of the dropwise addition, the reaction was continued at -78°C for 4 h. After completion of the reaction as monitored by TLC, 300 mL of a saturated ammonium chloride solution was added portionwise to the reaction solution to quench the reaction. The mixture was extracted twice with 200 mL of dichloromethane each time. The organic phases were combined, washed once with 200 mL of saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. 4.91 g of compound 20a was isolated by flash column chromatography (PE:EA=2:1, v:v) as a white solid in an 84% yield. 1 H NMR(400MHz, CDCl3) δ8.62(d,J=2.3Hz,1H),7.80(dt,J=8.4,1.9Hz,1H),7.23(d,J=8.3Hz,1H) ,4.62(d,J=6.0Hz,1H),4.59–4.53(m,1H),1.51(dd,J=6.8,1.2Hz,3H),1.27(d,J=1.3Hz,9H).

[0159] Synthesis of compound 21a:

[0160] The synthesis method of compound 21a refers to the synthesis of compound 14a, and compound 20a (4.9 g, 16.12 mmol) and 4-nitrophenylboronic acid (2.96 g, 17.73 mmol) are added to obtain 4.9 g of compound 21a as a yellow solid with a yield of 88%. 1 H NMR(400MHz, CDCl3)δ8.83(dt,J=2.3,1.0Hz,1H),8.41–8.31(m,2H),7.92(dd,J=8.1,2.4Hz,1H),7.78–7.72(m,2H ), 7.46 (d, J = 8.0Hz, 1H), 4.79 (d, J = 6.2Hz, 1H), 4.68 (p, J = 6.5Hz, 1H), 1.58 (d, J = 6.7Hz, 3H), 1.30 (d, J = 0.9Hz, 9H).

[0161] Synthesis of compound 22a:

[0162] 4.9 g of compound 21a was dissolved in a mixture of methanol and hydrochloric acid (40 mL of methanol, 25 mL of concentrated hydrochloric acid) and stirred at room temperature for 6 hours. After the reaction was completed as monitored by TLC, the methanol in the reaction solution was concentrated under reduced pressure. The residue was diluted with 20 mL of water, and a 2 mol / L sodium hydroxide solution was slowly added dropwise to adjust the pH to 10. The mixture was extracted with dichloromethane three times with 200 mL each time. The organic phases were combined and concentrated under reduced pressure. The resulting mixture was separated by flash column chromatography (DCM:MeOH=20:1, v:v) to obtain 3.16 g of compound 22a as a yellow solid in a yield of 92%. 1 H NMR(400MHz,DMSO- d6)δ8.91(dd,J=2.4,0.8Hz,1H),8.39–8.29(m,2H),8.18(dd,J=8.2,2.4Hz,1H),8.07–7.9 9(m,2H),7.64(d,J=8.2Hz,1H),4.08(q,J=6.7Hz,1H),2.09(s,2H),1.32(d,J=6.7Hz,3H).

[0163] Synthesis of compound 23a:

[0164] The synthesis method of compound 23a was similar to that of compound 15a. Compound 22a (0.6 g, 2.468 mmol) and compound 1a (0.59 g, 2.468 mmol) were added to obtain 635 mg of compound 23a as a yellow solid in a yield of 57.3%. ESI-MS m / z: 449.2.

[0165] Synthesis of compound 24a:

[0166] The synthesis method of compound 24a refers to the synthesis of compound 3a. Compound 23a (250 mg, 0.56 mmol), reduced iron powder (155 mg, 2.79 mmol) and ammonium chloride (149 mg, 2.79 mmol) were added to obtain 240 mg of compound 24a as a yellow solid with a yield of 95.3%.

[0167] Synthesis of compound I-24:

[0168] The synthesis method of compound I-24 refers to the synthesis of compound 4a. Compound 24a (200 mg, 0.48 mmol) and trans-3-(3-pyridyl)acrylic acid (106 mg, 0.72 mmol) were added to obtain 120 mg of compound I-24 as a yellow solid with a yield of 48.0%. 1H NMR (600MHz, DMSO-d6) δ10.44(s,1H),8.85(d,J=2.3Hz,1H),8.82(d,J=2.3Hz,1H),8.60(dd,J=4.8,1.6Hz,1H), 8.23(s,1H),8.08(dt,J=8.0,2.0Hz,1H),8.01(d,J=8.3Hz,1H),7.97-7.88(m,1H),7.84(d,J=8.6Hz,2H),7.71-7 .65(m,3H),7.51(dd,J=7.9,4.7Hz,1H),7.35(d,J=8.0Hz,1H),7.26(d,J=5.7Hz,1H),6.96(d,J=15.8Hz,1H),5.0 1(s,1H),4.49(s,1H),4.36-4.19(m,2H),1.52(d,J=7.2Hz,3H),0.95(d,J=7.0Hz,1H),0.53(s,3H),0.42(s,3H). 13 C NMR(126MHz,DMSO-d6)δ163.10,160.68,158.22,156.88,154.28,151.04,150.1 5,149.03,146.33,146.06,138.96,136.90,134.35,134.28,133.13,131.93,130 .52,128.77,127.02,124.11,124.05,120.64,119.77,62.72,57.86,51.83,48.5 1,39.94,39.77,39.61,39.44,39.27,39.11,38.94,26.78,21.43,17.68,13.59. ESI-HRMS m / z:550.2555[M+H] + .

[0169] Route 2:

[0170] Synthesis of compound 25a:

[0171] The synthesis method of compound 25a was similar to that of compound 14a. 2-acetyl-5-bromopyridine (2 g, 10 mmol), 4-nitrophenylboronic acid (1.83 g, 11 mmol), tetrakistriphenylphosphine palladium (0.57 g, 0.5 mmol), and cesium carbonate (6.5 g, 20 mmol) were added to give 2.5 g of crude compound 25a as a yellow solid. ESI-MS m / z: 243.1 [M+H] + .

[0172] Synthesis of compound 26a:

[0173] Compound 25a (3 g, 12.38 mmol), ammonium acetate (14.3 g, 185.77 mmol), and sodium cyanoborohydride (3.11 g, 49.54 mmol) were dissolved in 50 mL of anhydrous ethanol and heated to 120°C under nitrogen. The reaction mixture was refluxed for 10 min. After completion of the reaction, the reaction solution was cooled to room temperature and concentrated under reduced pressure to remove the solvent. 100 mL of ethyl acetate and 50 mL of water were added to dissolve the mixture, and the pH was adjusted to approximately 10. The mixture was stirred at room temperature for 15 min and extracted with ethyl acetate three times with 120 mL each. The combined organic phases were washed with water and then with saturated sodium chloride solution (60 mL each), dried over anhydrous sodium sulfate, and separated by flash column chromatography (DCM:MeOH = 15:1, v:v) to obtain 1.7 g of compound 26a as a yellow solid in a 56.67% yield. 1 H NMR (400MHz, CDCl3) δ8.82(d,J=2.0Hz,1H),8.35(d,J=8.7Hz,2H),7.92(dd,J=8.1,2.3Hz,1H ), 7.74 (d, J = 8.7Hz, 2H), 7.47 (d, J = 8.1Hz, 1H), 4.32 (q, J = 6.7Hz, 1H), 1.52 (d, J = 6.7Hz, 3H).

[0174] Synthesis of compound 23a:

[0175] Compound 26a (0.6 g, 2.468 mmol), compound 1a (0.59 g, 2.468 mmol), and DIPEA (1.6 g, 12.34 mmol) were dissolved in 15 mL of DMSO and heated to 110°C under nitrogen for 16 h. The reaction solution was cooled to room temperature, and 50 mL of ethyl acetate and 50 mL of water were added. The mixture was stirred at room temperature for 10 min, and extracted with ethyl acetate three times with 100 mL each. The organic phases were combined and washed successively with water and saturated sodium chloride aqueous solution once with 50 mL each. The organic phase was dried over anhydrous sodium sulfate and chiral separation was performed by flash column chromatography (PE:EA=5:1, v:v) to give 200 mg of compound 23a as a yellow solid in an 18% yield.

[0176] Example 14

[0177] Synthesis of compound I-25:

[0178] The synthesis method of compound I-25 refers to the synthesis of compound 4a, and compound 24a (176 mg, 0.42 mmol) and imidazo[1,2-a]pyridine-6-carboxylic acid (102 mg, 0.63 mmol) are added to obtain 198 mg of compound I-25 as a yellow solid with a yield of 89.6%. 1 H NMR (300MHz, DMSO-d6) δ10.49(s,1H),9.29(s,1H),8.83(d,J=2.3Hz,1H),8.23(d, J=4.0Hz,1H),8.13(s,1H),8.01(dd,J=8.2,2.4Hz,1H),7.90(d,J=8.7Hz,2H),7.80 -7.67(m,6H),7.34(d,J=7.0Hz,1H),7.24(d,J=5.7Hz,1H),4.99(s,1H),4.49(s,1H),4. 37-4.17(m,2H),1.52(d,J=7.1Hz,3H),1.24(d,J=5.2Hz,1H),0.53(s,3H),0.42(s,3H). 13 C NMR(126MHz,DMSO-d6)δ165.87,163.37,161.18,158.92,156.67,154.32,151.0 0,146.53,144.34,138.73,134.04,132.95,132.37,131.17,128.96,128.75,126 .83,123.55,120.75,120.27,118.82,115.92,114.41,62.69,57.82,54.81,51.9 1,39.96,39.79,39.62,39.46,39.29,39.12,38.96,26.78,21.46,17.69,13.59. ESI-HRMS m / z:563.2521[M+H] + .

[0179] Example 15

[0180] Synthesis of compound I-26:

[0181] The synthesis method of compound I-26 refers to the synthesis of compound 4a, and compound 24a (150 mg, 0.36 mmol) and 3-pyridyloxyacetic acid (83 mg, 0.54 mmol) are added to obtain 118 mg of compound I-26 as a light yellow solid with a yield of 65%. 1H NMR(500MHz,DMSO-d6)δ10.30(s,1H),8.80(s,1H),8.40(d,J=3.0Hz,1H),8.27–8.19(m, 2H),7.99(dd,J=8.3,2.4Hz,1H),7.84(s,1H),7.80-7.74(m,2H),7.68(d,J=8.4Hz,2H), 7.48-7.43(m,1H),7.38(dd,J=8.5,4.6Hz,1H),7.25(d,J=5.6Hz,1H),4.99(s,1H),4.85 (s,2H),4.48(s,1H),4.38-4.17(m,2H),1.52(d,J=7.2Hz,3H),0.51(s,3H),0.41(s,3H). 13 C NMR(126MHz,DMSO-d6)δ166.74,163.55,161.76,159.52,157.17,154.87, 154.65,147.10,142.78,138.67,138.53,134.68,133.44,132.86,127.46 ,124.61,121.72,120.69,119.32,67.57,63.21,55.39,40.59,40.50,40. 42,40.33,40.26,40.17,40.00,39.83,39.67,39.50,22.03,18.26,14.11. ESI-HRMS m / z:554.2518[M+H] + .

[0182] Example 16

[0183] Synthesis of compound I-34:

[0184] The synthesis method of compound I-34 refers to the synthesis of compound 4a, and compound 16a (200 mg, 0.48 mmol) and nicotinic acid (88.46 mg, 0.72 mmol) are added to obtain 167 mg of compound I-34 as a light yellow solid powder with a yield of 66.8%. 1H NMR(500MHz,DMSO-d6)δ10.85(s,1H),9.33(d,J=1.5Hz,1H),8.95(d,J=2.4Hz,1H) ,8.83(t,J=2.0Hz,1H),8.39(s,1H),8.27-8.13(m,2H),8.00(d,J=8.4Hz,2H),7.65 (dd,J=17.0,8.1Hz,4H),7.39(dd,J=19.1,7.1Hz,3H),5.08(s,1H),4.64(s,1H),4. 36(d,J=13.8Hz,2H),1.78(s,1H),1.51(d,J=7.0Hz,3H),0.70(s,3H),0.49(s,3H). 13 CNMR(126MHz,DMSO-d6)δ164.52,161.64,159.45,159.33,157.22,154.89 ,152.61,149.16,145.48,138.58,138.23,136.11,135.95,131.05,127.16 ,126.64,126.57,126.54,123.99,121.15,58.38,55.38,40.59,40.50,40. 42,40.33,40.16,40.00,39.83,39.66,39.50,27.34,23.87,18.29,14.15. ESI-HRMS m / z:523.2463[M+H] + .

[0185] Example 17

[0186] Synthesis of compound I-35:

[0187] The synthesis method of compound I-35 refers to the synthesis of compound 4a, and compound 16a (240 mg, 0.57 mol) and 2-carboxylic acid pyrazine (106 mg, 0.86 mmol) are added to obtain 240 mg of compound I-35 as a light yellow solid with a yield of 80%. 1H NMR(500MHz,DMSO-d6)δ10.52(s,1H),9.13(d,J=2.3Hz,1H),8.82-8.74(m,1H),8.31 (dt,J=8.0,2.0Hz,1H),8.19(s,1H),7.86(d,J=8.3Hz,2H),7.65(d,J=8.3Hz,2H),7. 62-7.57(m,3H),7.39(d,J=7.8Hz,2H),7.21(d,J=5.7Hz,1H),5.00(s,1H),4.61(s,1 H), 4.42-4.23 (m, 2H), 1.77 (s, 1H), 1.47 (d, J = 7.0Hz, 3H), 0.67 (s, 3H), 0.46 (s, 3H). 13 C NMR (126MHz, DMSO-d6) δ165.06,162.17,158.67,158.31,158.20,154.88,154.66,148.21,145.51,144.55,144.34,143.71,138.54,137.99, 136.26,127.20,126.81,126.55,121.41,58.73,54.08,40.49,40.41, 40.32,40.15,39.99,39.82,39.65,39.49,27.42,23.58,18.54,17.18. ESI-HRMS m / z:524.2412[M+H] + .

[0188] Example 18

[0189] Synthesis of compound I-36:

[0190] The synthesis method of compound I-36 refers to the synthesis of compound 4a, and compound 16a (200 mg, 0.48 mol) and furano[3,2-b]pyridine-2-carboxylic acid (116 mg, 0.72 mmol) are added to obtain 77 mg of compound I-36 as a white solid with a yield of 28.5%. 1H NMR(500MHz,DMSO-d6)δ10.63(s,1H),8.19(s,1H),7.96-7.90(m,2H),7.84(d,J=7.9H z,1H),7.80(s,1H),7.74(d,J=8.4Hz,1H),7.65(d,J=8.3Hz,2H),7.60(d,J=8.0Hz,2H) ,7.54-7.49(m,1H),7.38(t,J=7.5Hz,3H),7.22(d,J=5.7Hz,1H),5.01(s,1H),4.61(s ,1H),4.41-4.21(m,2H),1.77(s,1H),1.47(d,J=7.1Hz,3H),0.67(s,3H),0.47(s,3H). 13 C NMR (126MHz, DMSO-d6) δ162.77,161.58,159.42,157.10,154.95,149.26,145. 50,138.24,138.10,136.22,131.99,131.92,129.27,127.66,127.63,127.18, 126.66,124.35,123.41,121.33,112.43,111.22,58.38,54.07,40.59,40.50, 40.42,40.33,40.16,39.99,39.83,39.66,39.49,27.34,23.89,18.55,18.28. ESI-HRMS m / z:562.2455[M+H] + .

[0191] Example 19

[0192] Synthesis of compound 10a:

[0193] The synthesis method of compound 10a was similar to that of compound 1a. 2,4-dichloropyrimidine (1.00 g, 6.71 mmol) and (S)-4-phenyl-oxazolidin-2-one (927 mg, 6.71 mmol) were added to obtain 860 mg of compound 10a as a white solid in an 85.9% yield. ESI-HRMS m / z: 276.0508 [M+H] + .

[0194] Synthesis of compound I-37:

[0195] The synthesis of compound I-37 was carried out by referring to the synthetic route of compound I-17, and finally compound 40a (70 mg, 0.155 mol) and trans-3-(3-pyridyl)acrylic acid (34 mg, 0.23 mmol) were added to obtain 36 mg of compound I-37 as a yellow solid with a yield of 46%. 1 H NMR (600MHz, DMSO-d6) δ10.39(s,1H),8.85(d,J=2.3Hz,1H),8.60(dd,J=4.8,1.6Hz,1H ),8.19-8.12(m,1H),8.06(dt,J=8.1,1.9Hz,1H),7.80(d,J=8.2Hz,2H),7.71-7.58(m, 4H),7.50(dd,J=7.9,4.7Hz,1H),7.47-7.13(m,9H),7.05(s,1H),6.96(d,J=15.8Hz,1H ), 5.89 (s, 1H), 4.94 (s, 1H), 4.84 (t, J = 8.5Hz, 1H), 4.22 (s, 1H), 1.41 (d, J = 6.9Hz, 3H). 13 C NMR(101MHz,DMSO-d6)δ167.76,163.64,161.50,157.09,154.94,150.94,149 .82,141.15,140.95,138.94,138.34,137.44,135.72,135.10,134.68,131.10 ,129.35,128.24,127.40,127.07,126.52,124.74,124.60,124.44,121.86,1 20.23,71.05,57.52,40.69,40.48,40.28,40.07,39.86,39.65,39.44,23.09. ESI-HRMS m / z:583.2454[M+H] + .

[0196] Example 20

[0197] Synthesis of compound I-40:

[0198] The synthesis method of compound I-40 refers to the synthesis route 2 of compound I-24, and compound 46a (150 mg, 0.33 mmol) is added to obtain 174 mg of compound I-40 as a light yellow solid powder with a yield of 95%. 1H NMR (300MHz, DMSO-d6) δ10.34(s,1H),9.06(d,J=1.8Hz,2H),8.39(d,J=2.9Hz,1H),8.22(dd,J=4.6 ,1.4Hz,1H),8.19(d,J=1.5Hz,1H),7.79(d,J=3.3Hz,4H),7.74(s,1H),7.48–7.42(m,1H),7.37(dd ,J=8.5,4.5Hz,1H),7.23(d,J=5.8Hz,1H),5.13–4.92(m,1H),4.85(s,2H),4.55–4.08(m,3H),2.55 (s, 1H), 1.55 (t, J = 7.7Hz, 3H), 0.96 (d, J = 7.0Hz, 2H), 0.76 (d, J = 6.9Hz, 2H), 0.46 (d, J = 31.3Hz, 2H). 13 C NMR(126MHz,DMSO-d6)δ166.84,155.02,154.97,154.85,154.65,142.71,139.35,138.44,129.48, 127.66,127.53,124.64,121.85,120.73,120.65,67.58,63.33,58.60,21.07,18.24,18.12,14.41. ESI-HRMS m / z:577.2277[M+Na] + .

[0199] Example 21

[0200] Synthesis of compound I-41:

[0201] The synthesis method of compound I-41 refers to the synthesis of compound I-37, and compound 49a (250 mg, 0.5 mmol) is added to obtain 200 mg of light yellow solid I-37 with a yield of 86%. 1H NMR (400MHz, DMSO-d6) δ10.34(s,1H),8.83(d,J=2.2Hz,1H),8.59(dd,J=4.8,1.6Hz,1H),8.22( s,1H),8.05(dt,J=8.1,2.0Hz,1H),7.87(s,1H),7.81–7.59(m,4H),7.49(dd,J=8.0,4.8Hz,2H), 7.42(s,4H),7.20(d,J=6.7Hz,4H),6.94(d,J=15.9Hz,2H),5.15(s,1H),4.99(tt,J=8.0,3.0Hz ,1H),4.38(t,J=8.5Hz,1H),4.16(s,1H),3.28–2.93(m,1H),2.68(s,1H),1.51(d,J=7.0Hz,3H). 13 C NMR(101MHz,DMSO-d6)δ163.54,161.77,159.48,157.27,154.37,150.87,149.76,145.20,138.80,138.38,137.37,136.27,1 35.66,134.61,131.01,129.84,128.77,127.32,127.08,126.68,124.63,124.52,120.02,97.91,66.31,36.39,31.44,23.74. ESI-HRMS m / z:597.2603[M+H] + .

[0202] Example 22

[0203] Synthesis of compound I-42:

[0204] The synthesis method of compound I-42 refers to the synthesis of compound I-41, and compound 49a (250 mg, 0.5 mmol) is added to obtain 230 mg of compound I-42 as a white solid with a yield of 76%. 1H NMR (300MHz, DMSO-d6) δ10.20 (s, 1H), 8.38 (d, J = 2.9Hz, 1H), 8.21(dd,J=4.5,1.4Hz,2H),7.90(s,1H),7.64(s,3H),7.45(dd,J=2.9,1.4H z,2H),7.43–7.39(m,3H),7.36(dd,J=8.6,4.7Hz,2H),7.20(d,J=5.8Hz,4H) ,6.90(s,1H),5.14(s,1H),4.98(tt,J=7.7,2.9Hz,1H),4.82(s,2H),4.38(t ,J=8.5Hz,1H),4.15(s,1H),3.11(s,1H),2.65(s,1H),1.50(d,J=7.0Hz,3H). 13 C NMR (126MHz, DMSO-d6) δ166.57,161.78,157.28,154.65,154.37,145.23,142.76,138.53,138.31,137.9 7,136.28,135.91,128.77,127.22,127.08,126.70,124.59,121.73,120.42,67.59,66.32,55.38,23.67. ESI-HRMS m / z:601.2555[M+H] + .

[0205] Example 23

[0206] Synthesis of compound I-43:

[0207] The synthesis method of compound I-43 refers to the synthesis of I-41, and compound 49a (250 mg, 0.5 mmol) is added to obtain 220 mg of compound I-43 as a white solid with a yield of 73%. 1H NMR(400MHz,DMSO-d6)δ10.42(s,1H),9.28(s,1H),8.23(s,1H),8.13(s,1H),7.91 (s,1H),7.77(d,J=9.4Hz,3H),7.73–7.66(m,2H),7.43(s,5H),7.20(d,J=5.9Hz,5 H),6.91(s,1H),5.26–5.07(m,1H),5.00(dq,J=8.1,4.6,3.7Hz,1H),4.38(t,J=8. 5Hz, 1H), 4.17 (d, J = 13.1Hz, 1H), 3.18 (s, 1H), 2.68 (s, 1H), 1.52 (d, J = 7.0Hz, 3H). 13 C NMR (126MHz, DMSO-d6) δ163.84,161.79,157.29,154.38,145.25,145.01,138.62,138.36,136.29,135.96,13 4.85,129.88,129.42,128.78,127.16,126.72,123.89,121.03,120.73,116.56,114.89,66.32,55.38,23.68. ESI-HRMS m / z:610.2552[M+H] + .

[0208] Example 24

[0209] Synthesis of compound 51a:

[0210] Compound 51a was synthesized by referring to patent WO 2014 / 141104 A1.

[0211] Synthesis of compound I-44:

[0212] Iron powder (168 mg, 3 mmol) and ammonium chloride (161 mg, 3 mmol) were dissolved in 4 mL of water and 10 mL of acetone under nitrogen atmosphere and heated to 70°C for 30 min under reflux. Compound 53a (302 mg, 0.6 mmol) was dissolved in 10 mL of acetone and injected into the reaction mixture. The reaction was continued for 6 h before heating was stopped and the mixture was cooled to room temperature. The reaction mixture was filtered through celite, and the filter cake was rinsed with 50 mL of dichloromethane. The filtrate was concentrated under reduced pressure and dissolved in 100 mL of dichloromethane. The mixture was then washed with water and saturated brine (50 mL each time). The organic phase was concentrated under reduced pressure to remove the solvent and dissolved in 6 mL of anhydrous DMF. DIPEA (194 mg, 1.5 mmol) and trans-3-(3-pyridyl)acrylic acid (112 mg, 0.75 mmol) were added. The mixture was stirred in an ice bath for 10 min, followed by the addition of HATU (381 mg, 1 mmol). The ice bath was removed after 10 minutes, and the mixture was allowed to react at room temperature for 4 h. After the reaction was completed as monitored by TLC, 10 mL of ice water was added to the reaction solution to quench the reaction, and 10 mL of ethyl acetate was added and stirred for 10 minutes, followed by 100 mL of ethyl acetate to dissolve the mixture and transfer it to a separatory funnel. The mixture was washed with water, saturated sodium bicarbonate, and saturated brine once, 30 mL each time. The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. After flash column chromatography (DCM:MeOH=80:1 to DCM:MeOH=40:1, v:v), 220 mg of compound I-44 was obtained as a light yellow solid with a yield of 60%. 1 H NMR (400MHz, DMSO-d6) δ10.37(s,1H),8.84(d,J=2.2Hz,1H),8.60(dd,J=4.7,1.6Hz,1H),8.20(d,J=5 .7Hz,1H),8.06(dt,J=8.0,2.0Hz,1H),7.79(d,J=8.7Hz,2H),7.69–7.58(m,5H),7.49(dd,J=8.0,4.8 Hz,1H),7.45(d,J=8.4Hz,2H),7.24(d,J=5.7Hz,1H),6.95(d,J=15.8Hz,1H),5.33–5.21(m,1H),4.75 –4.67(m,1H),4.46–4.37(m,2H),4.27(s,1H),1.52(d,J=6.9Hz,3H),1.07(s,9H),0.90–0.79(m,3H). 13C NMR(101MHz,DMSO-d6)δ163.56,157.61,155.04,150.89,149.78,138.93,138.49,137.39,135.53,134.60, 131.00,127.37,127.05,126.61,124.63,124.54,120.13,74.00,64.66,63.63,56.96,55.39,28.51,16.20. ESI-HRMS m / z:607.3024[M+H] + .

[0213] Example 25

[0214] Synthesis of compound I-45:

[0215] The synthesis method of compound I-45 refers to the synthesis of compound I-44, replacing trans-3-(3-pyridyl)acrylic acid with 3-pyridyloxyacetic acid (115 mg, 0.75 mmol) to obtain 230 mg of compound I-45 as a light yellow solid powder with a yield of 62%. 1 H NMR(400MHz,DMSO-d6)δ10.25(s,1H),8.39(d,J=2.9Hz,1H),8.24–8.15(m,2H),7.7 2(d,J=8.8Hz,2H),7.65–7.56(m,4H),7.44(d,J=8.8Hz,3H),7.37(dd,J=8.4,4.5Hz, 1H),7.24(d,J=5.6Hz,1H),5.26(s,1H),4.83(s,2H),4.71(dt,J=7.2,4.9Hz,1H),4. 50–4.36(m,2H),4.26(s,1H),1.52(d,J=6.9Hz,3H),1.31–0.92(m,9H),0.84(s,3H). 13 C NMR (126MHz, DMSO-d6) δ166.61,155.04,154.65,142.77,138.53,138.42,138.10,135.77,127. 27,127.04,126.63,124.59,121.73,120.54,74.00,67.59,64.69,63.64,56.98,28.52,16.21. ESI-HRMS m / z:611.2974[M+H] + .

[0216] Example 26

[0217] Synthesis of compound I-46:

[0218] The synthesis method of compound I-46 refers to the synthesis of compound I-44, replacing trans-3-(3-pyridyl)acrylic acid with imidazo[1,2-a]pyridine-6-carboxylic acid (122 mg, 0.75 mmol) to obtain 215 mg of compound I-46 as a light yellow solid powder with a yield of 61%. 1 HNMR(400MHz,DMSO-d6)δ10.46(s,1H),9.29(s,1H),8.20(d,J=5.7Hz,1H),8.13( s,1H),7.86(d,J=8.7Hz,2H),7.78(dd,J=9.5,1.8Hz,1H),7.74–7.59(m,6H),7.46 (d,J=8.2Hz,2H),7.24(d,J=5.7Hz,1H),5.28(s,1H),4.77–4.65(m,1H),4.53–4. 36(m,2H),4.27(s,1H),1.53(d,J=6.9Hz,3H),1.06(s,9H),0.85(d,J=5.7Hz,3H). 13 C NMR(101MHz,DMSO-d6)δ163.91,157.61,155.05,145.09,138.76,138.46,135.80,135.06,129.42,127.20, 127.06,126.65,123.73,121.13,120.64,116.65,114.87,74.00,64.67,63.64,56.97,55.39,28.51,16.20. ESI-HRMS m / z:620.2969[M+H] + .

[0219] Example 27

[0220] Synthesis of compound I-47:

[0221] Compound I-44 (150 mg, 0.247 mmol) was dissolved in 1 mL of anhydrous dichloromethane, 2 mL of trifluoroacetic acid was added, and the mixture was stirred at room temperature for 16 hours. After the reaction was completed by TLC monitoring, 10 mL of ethyl acetate was added to the reaction solution, and saturated sodium bicarbonate solution was slowly added dropwise under vigorous stirring until no bubbles were generated. Extracted with 50 mL of ethyl acetate, the organic phase was washed once with saturated sodium bicarbonate solution and saturated brine, each time 20 mL each. The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. After flash column chromatography (DCM:MeOH=50:1-20:1, v:v), 125 mg of compound I-47 was obtained as a yellow solid with a yield of 92%. 1 H NMR (400MHz, DMSO-d6) δ10.39(s,1H),8.84(d,J=2.2Hz,1H),8.60(dd,J=4.8,1.6Hz,1H),8.1 5(d,J=5.6Hz,1H),8.06(d,J=8.1Hz,1H),7.89(s,1H),7.79(d,J=8.7Hz,2H),7.69–7.56(m,5 H),7.49(dt,J=10.0,5.1Hz,3H),7.17(d,J=5.5Hz,1H),6.96(d,J=15.8Hz,1H),5.17–4.95(m ,2H),4.84–4.75(m,1H),4.51–4.35(m,2H),4.03(s,1H),1.47(d,J=7.0Hz,3H),0.65(s,3H). 13 C NMR (126MHz, DMSO-d6) δ163.57,157.40,155.01,150.87,149.75,145.31,138.86,138.44,137.37,135.7 5,134.61,131.03,127.40,127.02,126.63,124.68,124.52,120.14,64.06,63.73,57.46,50.13,23.52. ESI-HRMS m / z:551.2398[M+H] + .

[0222] Example 28

[0223] Synthesis of compound I-48:

[0224] The synthesis method of compound I-48 refers to the synthesis of compound I-47, replacing compound I-44 with compound I-45 (151 mg, 0.247 mmol) to obtain 126 mg of compound I-48 as a white solid powder with a yield of 92%. 1H NMR (400MHz, DMSO-d6) δ10.22(s,1H),8.39(d,J=2.9Hz,1H),8.22(dd,J=4.5,1.3Hz,1H),8.15 (d,J=5.7Hz,1H),7.85(s,1H),7.71(d,J=8.7Hz,2H),7.65–7.55(m,4H),7.51–7.41(m,3H),7.3 7(dd,J=8.4,4.6Hz,1H),7.17(d,J=5.7Hz,1H),5.10(s,1H),4.98(s,1H),4.83(s,2H),4.79(d dd,J=8.5,4.0,2.8Hz,1H),4.49–4.37(m,2H),4.03(s,1H),1.47(d,J=7.0Hz,3H),0.65(s,3H). 13 C NMR (126MHz, DMSO-d6) δ166.60,161.59,155.00,154.65,145.36,142.76,138.53,138.36,138.03,1 35.98,127.30,127.01,126.64,124.60,121.73,120.52,67.60,64.04,63.72,57.45,50.12,23.52. ESI-HRMS m / z:555.2346[M+H] + .

[0225] Example 29

[0226] Synthesis of compound I-49:

[0227] The synthesis method of compound I-49 refers to the synthesis of compound I-47, replacing compound I-44 with compound I-46 (153 mg, 0.247 mmol) to obtain 127 mg of compound I-49 as a white solid powder with a yield of 92%. 1H NMR (400MHz, DMSO-d6) δ10.43(s,1H),9.28(s,1H),8.21–8.08(m,2H),7.85(d,J=8.7Hz,2H ),7.77(dd,J=9.5,1.8Hz,1H),7.73–7.64(m,4H),7.61(d,J=8.1Hz,2H),7.48(d,J=7.9Hz,2 H),7.18(d,J=5.7Hz,1H),5.11(s,1H),4.99(s,1H),4.83–4.76(m,1H),4.47(dd,J=9.0,2. 8Hz, 1H), 4.41 (t, J = 8.7Hz, 1H), 3.98 (d, J = 56.8Hz, 1H), 1.48 (d, J = 7.0Hz, 3H), 0.67 (s, 3H). 13 C NMR(126MHz,DMSO-d6)δ163.91,157.39,155.01,145.36,145.10,138.68,138.40,136.03,135.04,129.40,1 27.24,127.03,126.66,123.75,121.14,120.68,116.64,114.86,64.06,63.73,57.46,50.13,49.08,23.52. ESI-HRMS m / z:564.2349[M+H] + .

[0228] Example 30

[0229] Synthesis of compound 55a:

[0230] Compound 55a was synthesized by referring to patent WO 2014 / 141104 A1.

[0231] Synthesis of compound I-50:

[0232] The synthesis method of compound I-50 refers to the synthesis of compound I-44, and compound 57a (160 mg, 0.35 mmol) is added to obtain 170 mg of compound I-50 as a light yellow solid with a yield of 86%. 1H NMR (400MHz, DMSO-d6) δ10.37(s,1H),8.83(d,J=2.2Hz,1H),8.59(dd,J=4.8,1.6Hz,1H),8.21(s,1H), 8.05(d,J=8.1Hz,1H),7.92(s,1H),7.81–7.75(m,2H),7.68–7.57(m,5H),7.49(dd,J=8.0,4.8Hz,1H), 7.38(d,J=7.7Hz,2H),7.22(d,J=5.6Hz,1H),6.95(d,J=15.8Hz,1H),4.96(s,1H),4.73(dd,J=30.0,8. 1Hz, 1H), 4.38 (d, J = 49.4Hz, 3H), 1.46 (d, J = 7.0Hz, 3H), 1.24 (d, J = 6.7Hz, 1H), 1.08 (d, J = 23.6Hz, 2H). 13 C NMR (126MHz, DMSO-d6) δ163.57,161.51,159.68,154.51,150.87,149.76,138.90,138.31,137.38,135.56,134.61, 131.02,127.34,126.66,126.37,124.66,124.52,120.17,97.50,89.01,87.65,62.29,50.33,23.99,16.79,16.63. ESI-HRMS m / z:575.2173[M+Na] + .

[0233] Example 31

[0234] Synthesis of compound I-51:

[0235] The synthesis method of compound I-51 refers to the synthesis of compound I-45, and compound 57a (160 mg, 0.35 mmol) is added to obtain 188 mg of compound I-51 as a light yellow solid with a yield of 97%. 1H NMR (400MHz, DMSO-d6) δ10.22(s,1H),8.38(d,J=3.0Hz,1H),8.21(d,J=4.4H z,2H),7.91(s,1H),7.71(d,J=8.5Hz,2H),7.65–7.55(m,4H),7.46–7.32(m, 4H),7.22(d,J=5.6Hz,1H),4.96(s,1H),4.82(s,2H),4.72(dd,J=30.6,7.7H z, 1H), 4.38 (d, J = 49.9Hz, 3H), 1.45 (d, J = 7.0Hz, 3H), 1.07 (d, J = 25.0Hz, 3H). 13 C NMR (126MHz, DMSO-d6) δ166.61,161.52,159.69,156.91,154.65,154.51,145.36,142.77,138.53,138.23,138.08, 135.79,127.25,126.68,126.36,124.59,121.72,120.55,97.49,67.59,62.27,55.38,50.33,23.99,16.79,16.62. ESI-HRMS m / z:579.2120[M+Na] + .

[0236] Example 32

[0237] Synthesis of compound I-52:

[0238] The synthesis method of compound I-52 refers to the synthesis of compound I-46, and compound 57a (50 mg, 0.35 mmol) is added to obtain 55 mg of compound I-52 as a light yellow solid with a yield of 88%. 1H NMR(400MHz,DMSO-d6)δ10.46(s,1H),9.29(s,1H),8.22(s,1H),8.13(s,1H),7.96(s,1H),7.86(d,J =8.8Hz,2H),7.78(dd,J=9.5,1.8Hz,1H),7.73–7.70(m,1H),7.69(s,1H),7.66(s,1H),7.64(d,J=7. 9Hz,2H),7.49–7.34(m,2H),7.23(d,J=5.6Hz,1H),5.05(d,J=58.8Hz,1H),4.73(dd,J=30.5,8.4Hz, 1H), 4.45 (s, 2H), 4.26 (d, J = 56.6Hz, 1H), 1.46 (d, J = 7.0Hz, 3H), 1.24 (s, 1H), 1.10 (d, J = 23.3Hz, 2H). 13 C NMR (126MHz, DMSO-d6) δ163.84,154.52,144.90,138.72,138.28,135.86,134.60,129.49,127.18,1 26.70,126.37,124.11,121.17,120.85,116.47,114.94,97.51,62.29,55.38,23.99,16.80,16.63. ESI-HRMS m / z:566.2305[M+H] + .

[0239] Example 33

[0240] Synthesis of compound I-53:

[0241] The synthesis method of compound I-53 refers to the synthesis of I-44, and compound 59a (333 mg, 0.66 mmol) is added to obtain 360 mg of compound I-53 as a light yellow solid powder with a yield of 90%. 1H NMR (400MHz, DMSO-d6) δ10.45(s,1H),8.84(dd,J=8.7,2.3Hz,2H),8.60(dd,J=4.7,1.6Hz ,1H),8.22(d,J=5.1Hz,1H),8.10–7.98(m,2H),7.84(d,J=8.5Hz,2H),7.79–7.67(m,3H), 7.65(s,1H),7.53–7.42(m,2H),7.26(d,J=5.7Hz,1H),6.96(d,J=15.9Hz,1H),5.29(s,1H ), 4.68 (s, 1H), 4.43 (s, 2H), 4.28 (s, 1H), 1.56 (d, J = 7.0Hz, 3H), 1.06 (s, 9H), 0.77 (s, 3H). 13 C NMR (126MHz, DMSO-d6) δ163.67,161.94,159.42,157.63,155.04,150.90,149.76,146.86,139.56,137.54,134.82,134.63,13 3.86,132.44,130.99,127.62,124.58,124.52,120.29,98.30,74.04,64.60,63.62,57.09,55.37,51.34,28.69,28.49,16.07. ESI-HRMS m / z:608.2968[M+H] + .

[0242] Example 34

[0243] Synthesis of compound I-54:

[0244] The synthesis method of compound I-54 refers to the synthesis of compound I-45, and compound 59a (333 mg, 0.66 mmol) is added to obtain 380 mg of compound I-54 as a white solid powder with a yield of 94%. 1H NMR (400MHz, DMSO-d6) δ10.29(s,1H),8.81(d,J=2.4Hz,1H),8.39(d,J=3.0Hz,1H),8.22(d,J= 4.8Hz,2H),8.03(dd,J=8.3,2.4Hz,1H),7.76(d,J=8.4Hz,2H),7.69(d,J=8.4Hz,2H),7.44(d, J=8.5Hz,2H),7.37(dd,J=8.5,4.6Hz,1H),7.26(d,J=5.7Hz,1H),5.28(s,1H),4.84(s,2H),4. 67(d,J=8.8Hz,1H),4.42(s,2H),4.27(s,1H),1.55(d,J=7.0Hz,3H),1.05(s,9H),0.77(s,3H). 13 C NMR(101MHz,DMSO-d6)δ166.74,162.02,157.62,155.04,154.63,146.88,142.76,138.74,138.51,134.86,133 .77,132.67,127.54,124.60,121.71,120.64,98.27,74.04,67.55,64.58,63.61,57.06,51.34,28.47,16.07. ESI-HRMS m / z:612.2926[M+H] + .

[0245] Example 35

[0246] Synthesis of compound I-55:

[0247] The synthesis method of compound I-55 refers to the synthesis of compound I-46, and compound 59a (333 mg, 0.66 mmol) is added to obtain 350 mg of compound I-55 as a white solid powder with a yield of 86%. 1H NMR (400MHz, DMSO-d6) δ10.50(s,1H),9.29(s,1H),8.85(d,J=2.3Hz,1H),8.23(d,J=6. 1Hz,1H),8.13(s,1H),8.07(dd,J=8.2,2.4Hz,1H),7.91(d,J=8.7Hz,2H),7.81–7.67(m, 5H),7.46(d,J=8.2Hz,1H),7.26(d,J=5.7Hz,1H),5.29(s,1H),4.68(dt,J=7.9,4.6Hz,1 H), 4.42 (d, J = 4.0Hz, 2H), 4.29 (s, 1H), 1.56 (d, J = 7.0Hz, 3H), 1.06 (s, 9H), 0.78 (s, 3H). 13 C NMR (126MHz, DMSO-d6) δ164.00,162.01,157.64,155.04,146.90,145.10,139.41,135.06,134.86,133.82,132. 73,129.46,127.46,123.72,121.26,120.60,116.66,114.87,74.05,64.62,63.62,57.09,51.36,28.50,16.08. ESI-HRMS m / z:621.2928[M+H] + .

[0248] Example 36

[0249] Synthesis of compound I-56:

[0250] The synthesis method of compound I-56 refers to the synthesis of compound I-47, and compound I-53 (260 mg, 0.42 mmol) is added to obtain 200 mg of compound I-56 as a light yellow solid powder with a yield of 86%. 1H NMR (400MHz, DMSO-d6) δ10.44(s,1H),8.83(dd,J=15.3,2.3Hz,2H),8.60(dd,J=4.8,1.6Hz,1H),8.18(s,1 H),8.06(d,J=8.1Hz,1H),8.00(dd,J=8.2,2.4Hz,1H),7.84(d,J=8.7Hz,2H),7.79–7.62(m,4H),7.50(dd,J =8.0,4.8Hz,1H),7.43(d,J=8.2Hz,1H),7.30–7.15(m,1H),6.96(d,J=15.8Hz,1H),5.12(t,J=1.6Hz,1H),4 .79(d,J=55.5Hz,2H),4.41(s,2H),3.77(s,1H),1.51(d,J=7.0Hz,3H),0.91(t,J=7.4Hz,1H),0.55(s,2H). 13 C NMR(126MHz,DMSO-d6)δ163.66,162.81,157.52,155.03,150.88,149.75,146.97,139.50,137.52,134.78,134.64, 133.78,131.00,129.13,127.66,124.60,124.53,120.26,65.49,64.11,63.82,57.50,51.90,30.47,19.12,14.01. ESI-HRMS m / z:552.2347[M+H] + .

[0251] Example 37

[0252] Synthesis of compound I-57:

[0253] The synthesis method of compound I-57 was based on the synthesis of compound I-47. Compound I-54 (280 mg, 0.46 mmol) was added to obtain 220 mg of compound I-57 as a white solid with a yield of 87%. 1H NMR (400MHz, DMSO-d6) δ10.33(s,1H),8.81(d,J=2.3Hz,1H),8.41(d,J=2.9Hz,1H),8.2 4(d,J=4.6Hz,1H),8.19(s,1H),8.03(dd,J=8.2,2.4Hz,1H),7.98–7.81(m,1H),7.80–7 .66(m,4H),7.52–7.38(m,3H),7.24(d,J=5.8Hz,1H),5.14(d,J=13.5Hz,1H),4.86(s,4 H), 4.41 (s, 2H), 3.75 (s, 1H), 1.52 (d, J = 7.0Hz, 3H), 0.97 (d, J = 6.1Hz, 1H), 0.56 (s, 2H). 13 C NMR(101MHz,DMSO-d6)δ166.70,162.49,157.71,154.98,154.78,146.66,142.32,138.78,138.05,1 35.26,133.90,132.65,127.62,124.82,122.31,120.59,67.55,64.07,63.88,57.52,51.79,21.94. ESI-HRMS m / z:556.2294[M+H] + .

[0254] Example 38

[0255] Synthesis of compound I-58:

[0256] The synthesis method of compound I-58 refers to the synthesis of compound I-47, and compound I-55 (240 mg, 0.42 mmol) is added to obtain 200 mg of compound I-58 as a white solid powder with a yield of 90%. 1H NMR (400MHz, DMSO-d6) δ10.50(s,1H),9.29(t,J=1.4Hz,1H),8.83(d,J=2.4Hz,1H),8.18(s,1H),8 .15–8.10(m,1H),8.02(dd,J=8.2,2.4Hz,1H),7.90(d,J=8.7Hz,2H),7.79(d,J=1.8Hz,1H),7.78–7 .74(m,2H),7.74–7.65(m,3H),7.44(d,J=8.0Hz,1H),7.21(s,1H),5.13(d,J=10.6Hz,1H),4.87(s, 1H), 4.72 (s, 1H), 4.41 (s, 2H), 3.78 (s, 1H), 1.52 (d, J = 7.0Hz, 3H), 1.07–0.90 (m, 1H), 0.56 (s, 2H). 13 C NMR (126MHz, DMSO-d6) δ164.00,162.91,155.04,147.04,145.10,139.33,135.05,134.79,133.73,129. 46,127.50,123.74,121.24,120.61,120.35,116.66,114.87,64.11,63.82,57.50,55.37,51.92,21.98. ESI-HRMS m / z:565.2298[M+H] + .

[0257] Example 39

[0258] Synthesis of compound I-59:

[0259] The synthesis method of compound I-59 refers to the synthesis of compound I-50, and compound 61a (135 mg, 0.30 mmol) is added to obtain 150 mg of compound I-59 as a light yellow solid powder with a yield of 90%. 1H NMR (400MHz, DMSO-d6) δ10.45(s,1H),8.84(dd,J=4.8,2.2Hz,2H),8.60(dd,J=4.8,1.6Hz,1H),8.26(d,J= 5.6Hz,1H),8.10–8.00(m,2H),7.96(d,J=7.2Hz,1H),7.88–7.80(m,2H),7.77–7.62(m,3H),7.50(dd,J=7.9 ,4.8Hz,1H),7.41–7.34(m,1H),7.25(d,J=5.6Hz,1H),6.96(d,J=15.8Hz,1H),4.97(s,1H),4.66(d,J=31. 2Hz,1H),4.42(s,2H),4.37–4.15(m,1H),1.51(d,J=7.1Hz,3H),1.23(s,1H),1.01(dd,J=23.8,6.4Hz,2H). 13 C NMR (126MHz, DMSO-d6) δ163.67,159.74,154.47,150.89,149.76,147.02,139.54,137.53,134.86,134. 64,133.67,132.45,131.00,127.58,124.59,124.52,120.31,89.08,87.72,62.33,22.01,16.85,16.68. ESI-HRMS m / z:554.2303[M+H] + .

[0260] Example 40

[0261] Synthesis of compound I-60:

[0262] The synthesis method of compound I-60 refers to the synthesis of compound I-51, and compound 61a (135 mg, 0.30 mmol) is added to obtain 160 mg of compound I-60 as a white solid with a yield of 95%. 1H NMR (400MHz, DMSO-d6) δ10.30(s,1H),8.82(s,1H),8.39(d,J=3.0Hz,1H),8.23(dd,J=14.1,5.1Hz,2H),7.99(dd,J=31.1,7.4Hz,2H),7.8 5–7.62(m,4H),7.44(dd,J=8.5,3.0Hz,1H),7.37(dd,J=8.4,4.3Hz,2H),7.24(d,J=5.7Hz,1H),4.97(s,1H),4.84(s,2H),4.65(d,J=30.6 Hz,1H),4.41(d,J=8.0Hz,2H),4.36–4.12(m,1H),1.50(d,J=7.1Hz,3H),1.24(s,1H),1.00(dd,J=24.1,6.3Hz,2H). 13 C NMR(101MHz,DMSO-d6)δ166.74,163.59,159.78,156.85,154.63,154.47,147.04,142.76,138.73,138.51,134.90,133 .57,132.67,127.49,124.61,121.71,120.66,119.29,97.61,89.25,67.54,62.32,57.75,52.50,22.02,16.87,16.66. ESI-HRMS m / z:558.2250[M+H] + .

[0263] Example 41

[0264] Synthesis of compound I-61:

[0265] The synthesis method of compound I-61 refers to the synthesis of compound I-52, and compound 61a (135 mg, 0.30 mmol) is added to obtain 150 mg of compound I-61 as a white solid with a yield of 88%. 1H NMR (400MHz, DMSO-d6) δ10.51(s,1H),9.34–9.26(m,1H),8.86(d,J=2.4Hz,1H),8.26(d,J=5.6Hz,1 H),8.14(s,1H),8.06(d,J=8.3Hz,1H),7.96(d,J=7.3Hz,1H),7.93–7.85(m,2H),7.83–7.63(m,5H), 7.45–7.35(m,1H),7.25(d,J=5.7Hz,1H),4.98(s,1H),4.66(d,J=31.4Hz,1H),4.42(d,J=8.1Hz,2H) ,4.30(d,J=53.0Hz,1H), 1.51(d,J=7.1Hz,3H), 1.24(d,J=6.9Hz,1H), 1.02(dd,J=24.2,6.3Hz,2H). 13 C NMR(126MHz,DMSO-d6)δ163.97,154.48,147.06,145.00,139.37,134.90,134.83,133.63,132.75,129.49,127.42,12 3.92,121.28,120.71,119.34,116.57,114.92,97.64,89.10,87.74,62.35,57.76,55.36,52.51,22.02,16.85,16.69. ESI-HRMS m / z:567.2255[M+H] + .

[0266] Example 42

[0267] Synthesis of compound I-62:

[0268] The synthesis steps of compound I-62 refer to compound I-15, and compound 66a (200 mg, 0.352 mmol) and trans-3-(3-pyridyl)acrylic acid (53 mg, 0.357 mmol) are added to obtain 86 mg of compound I-62 as a white solid with a yield of 49.8%. 1H NMR (300MHz, DMSO-d6) δ9.89(s,1H),8.74(d,J=2.2Hz,1H),8.54(d,J=4.8Hz,1H),8.27(t,J=5.8Hz,1 H),8.14(d,J=5.6Hz,1H),7.95(d,J=8.0Hz,1H),7.64(s,1H),7.58–7.37(m,4H),7.29(d,J=8.2Hz,2H) ,7.10(d,J=5.6Hz,1H),6.75(d,J=15.9Hz,1H),5.03(s,1H),4.79(s,1H),4.52(t,J=8.9Hz,1H),4.32 (s, 1H), 3.46 (q, J = 6.4Hz, 2H), 2.55 (d, J = 6.9Hz, 2H), 1.44 (d, J = 6.9Hz, 3H), 1.24 (m, 1H), 0.61 (s, 6H). 13 C NMR(75MHz,DMSO-d6)δ169.10,164.53,159.11,154.39,152.06,150.00,148.99,140.78,138.19,137.40,135.17,133.81,130.64,12 5.79,124.15,123.85,121.80,118.97,118.96,110.75,63.70,58.89,54.83,49.92,39.79,36.16,35.25,27.14,23.24,17.21,14.14. ESI-HRMS m / z:600.2383[M+H] + .

[0269] Example 43

[0270] Synthesis of compound I-63:

[0271] The synthesis steps of compound I-63 refer to the synthesis of compound I-16, and compound 68a (150 mg, 0.32 mmol) and trans-3-(3-pyridyl)acrylic acid (71 mg, 0.48 mol) are added to obtain 54 mg of compound I-63 as a light yellow solid powder with a yield of 27.3%. 1H NMR(400MHz,DMSO-d6)δ10.36(s,1H),8.79(s,1H),8.55(s,1H),8.11(s,1H ),8.00(d,J=7.3Hz,1H),7.74(d,J=7.2Hz,3H),7.66–7.34(m,8H),7.07(d,J =4.7Hz,1H),6.93(d,J=14.9Hz,1H),5.07(s,1H),4.74(s,1H),4.47(s,1H) ,4.29(s,1H),2.99(s,1H),1.45(d,J=5.6Hz,3H),0.71(s,3H),0.49(s,3H). 13 C NMR(100MHz,DMSO-d6)δ163.63,159.70,155.92,155.00,152.69,150.95,149.83,138.92,138.36,137.39,135.76,134.66,131 .09,127.38,126.70,124.76,124.59,122.37,120.20,111.41,64.31,59.50,59.46,55.45,46.12,27.71,23.92,17.83,14.70. ESI-HRMS m / z:605.2337[M+H] + .

[0272] Example 44

[0273] Synthesis of compound I-64:

[0274] The synthesis steps of compound I-64 refer to the synthesis of compound I-16, and compound 68a (150 mg, 0.32 mmol) and imidazo[1,2-a]pyridine-6-carboxylic acid (77 mg, 0.48 mmol) are added to obtain 71 mg of solid compound I-64 with a yield of 36.3%. 1H NMR (400MHz, DMSO-d6) δ10.38(s,1H),9.23(s,1H),8.11(d,J=5.5Hz,1H),8.08(s ,1H),7.81(s,1H),7.67–7.64(m,5H),7.59(d,J=8.6Hz,2H),7.55(d,J=8.2Hz,2H ),7.41(d,J=7.4Hz,2H),7.08(d,J=5.5Hz,1H),5.05(s,1H),4.72(s,1H),4.47(s ,1H),4.28(s,1H),1.45(d,J=7.0Hz,3H),1.19(s,1H),0.61(s,3H),0.46(s,3H). 13 C NMR(100MHz,DMSO-d6)δ163.98,159.76,157.45,155.01,154.45,152.68,145.16,144.65,138.72,138.30,136.05,135.31,1 35.14,129.46,127.23,126.74,123.77,121.19,120.71,116.73,114.93,111.41,64.31,59.47,55.46,27.70,17.83,14.68. ESI-HRMS m / z:618.2291[M+H] + .

[0275] Example 45

[0276] Synthesis of compound I-71:

[0277] The synthesis steps of compound I-71 refer to the synthesis of compound I-17, and compound 71a (130 mg, 0.274 mmol) and trans-3-(3-pyridyl)acrylic acid (61.4 mg, 0.412 mol) are added to obtain 106 mg of compound I-71 as a light yellow solid powder with a yield of 63%. 1H NMR (300MHz, DMSO-d6) δ10.38(s,1H),8.84(d,J=2.2Hz,1H),8.59(dd,J=4.8,1.6Hz,1H),8.16(d,J=5. 6Hz,1H),8.05(dt,J=7.7,1.8Hz,1H),7.78(d,J=8.6Hz,3H),7.67(s,1H),7.63-7.55(m,4H),7.49(dd,J =7.9,4.8Hz,1H),7.44(d,J=7.9Hz,2H),7.12(d,J=5.6Hz,1H),6.95(d,J=15.8Hz,1H),5.10(s,1H),4.7 8(s,1H),4.52(t,J=9.1Hz,1H),4.32(s,1H),1.49(d,J=7.1Hz,3H),1.25(d,J=4.5Hz,1H),0.50(s,6H). 13 C NMR(101MHz,DMSO-d6)δ163.62,159.76,155.01,152.69,150.94,149.89,149.82,139.07,138.97,138.89,138.35, 137.43,135.77,134.67,131.08,127.38,126.81,126.70,126.59,124.73,124.59,122.40,120.21,111.41,64.31, 59.46,40.69,40.48,40.27,40.06,39.85,39.64,39.43,27.71,17.82,14.70. ESI-HRMS m / z:605.2337[M+H] + .

[0278] Example 46

[0279] Synthesis of compound I-72:

[0280] The synthesis steps of compound I-72 refer to the synthesis of I-16, and compound 71a (300 mg, 0.63 mmol) and imidazo[1,2-a]pyridine-6-carboxylic acid (154 mg, 0.95 mmol) are added to obtain 226 mg of compound I-72 as a white solid with a yield of 76.3%. 1H NMR (300MHz, DMSO-d6) δ10.56(s,1H),9.38(s,1H),8.78(dd,J=4.4,1.4Hz,1H),8.55(dd,J=8.4,1.4Hz,1H), 8.28-8.24(m,1H),8.16(d,J=5.6Hz,1H),8.01(dd,J=9.5,1.8Hz,1H),7.93(d,J=1.6Hz,1H),7.84(d,J=8.8Hz ,3H),7.67-7.58(m,4H),7.53(dd,J=8.4,4.4Hz,1H),7.45(d,J=7.9Hz,2H),7.12(d,J=5.6Hz,1H),5.10(s,1H ), 4.77 (s, 1H), 4.52 (t, J = 8.6Hz, 1H), 4.33 (s, 1H), 1.50 (d, J = 7.0Hz, 3H), 1.24 (d, J = 5.1Hz, 1H), 0.50 (s, 6H). 13 C NMR(101MHz,DMSO-d6)δ163.30,159.74,155.01,152.71,151.66,143.39,140 .15,138.51,138.26,136.26,135.17,132.21,131.19,130.09,129.39,127.2 7,127.03,126.75,122.46,121.27,115.69,115.45,115.15,111.42,64.31,5 9.47,40.67,40.46,40.26,40.05,39.84,39.63,39.42,27.72,17.83,14.71. ESI-HRMS m / z:618.2293[M+H] + .

[0281] Example 47

[0282] Synthesis of compound I-73:

[0283] The synthesis method of compound I-73 refers to the synthesis of compound I-24, and compound 73a (150 mg, 0.32 mmol) and trans-3-(3-pyridyl)acrylic acid (71 mg, 0.48 mol) are added to obtain 140 mg of compound I-73 as a light yellow solid powder with a yield of 60.94%. 1H NMR (300MHz, DMSO-d6) δ10.43(s,1H),8.83(dd,J=9.9,2.3Hz,2H),8.60(dd,J=4.8,1.6Hz,1H),8.17(d, J=5.8Hz,1H),8.06(dt,J=8.1,1.9Hz,1H),7.97(dd,J=8.2,2.4Hz,1H),7.82(d,J=8.7Hz,2H),7.74-7.60 (m,4H),7.49(dd,J=8.0,4.8Hz,1H),7.41(d,J=8.3Hz,1H),7.14(d,J=5.5Hz,1H),6.95(d,J=15.8Hz,1H) ,5.11(s,1H),4.63(s,1H),4.54-4.24(m,2H),1.53(d,J=7.1Hz,3H),1.24(d,J=4.9Hz,1H),0.42(s,6H). 13 C NMR (126MHz, DMSO-d6) δ163.10,159.18,154.37,152.08,150.97,150.34,149. 20,146.47,138.90,138.37,136.96,134.15,134.07,133.03,132.04,130.44, 128.74,126.98,124.03,123.96,121.79,120.60,119.74,63.64,58.85,54.81 ,39.96,39.79,39.63,39.46,39.29,39.13,38.96,27.04,21.56,17.25,14.02. ESI-HRMS m / z:606.2288[M+H] + .

[0284] Example 48

[0285] Synthesis of compound I-74:

[0286] The synthesis method of compound I-74 refers to the synthesis of compound I-24, and compound 71a (179 mg, 0.38 mmol) and imidazo[1,2-a]pyridine-6-carboxylic acid (92 mg, 0.57 mmol) are added to obtain 130 mg of compound I-74 as a white solid with a yield of 66.3%. 1H NMR (300MHz, DMSO-d6) δ10.48(s,1H),9.28(t,J=1.4Hz,1H),8.83(d,J=2.3Hz,1H),8.20-8.11(m,2H),8.00(dd, J=8.2,2.4Hz,1H),7.89(d,J=8.7Hz,2H),7.78-7.72(m,3H),7.71(d,J=1.3Hz,2H),7.68(dt,J=3.3,1.3Hz,1H), 7.42(d,J=7.5Hz,1H),7.15(d,J=5.5Hz,1H),5.12(s,1H),4.73-4.44(m,2 H), 4.29 (s, 1H), 1.54 (d, J = 7.1Hz, 3H), 1.24 (d, J = 4.8Hz, 1H), 0.43 (s, 6H). 13 C NMR (126MHz, DMSO-d6) δ167.27,163.44,159.18,154.36,152.07,146.50,144. 55,138.74,134.52,134.18,132.98,132.32,131.54,131.34,128.88,128.60, 126.82,123.13,120.70,120.44,120.04,116.11,114.30,63.64,58.85,54.81 ,52.50,39.95,39.79,39.62,39.45,39.29,39.12,38.95,21.57,17.25,14.04. ESI-HRMS m / z:619.2222[M+H] + .

[0287] Example 49

[0288] Synthesis of compound 81a:

[0289] 2,4-Dichloro-7H-pyrrolo[2,3-d]pyrimidine (5.0 g, 26.60 mmol) and p-toluenesulfonyl chloride (5.58 g, 29.26 mmol) were dissolved in 25 mL of anhydrous DMF and stirred in an ice bath for 5 minutes. Then, a suspension of 60% NaH (1.17 g, 29.26 mmol) in DMF (10 mL) was slowly added dropwise. After the addition was completed, the mixed solution was transferred to room temperature and stirred for 2 h. After the reaction was completed as monitored by TLC, 100 mL of ethyl acetate was added to the reaction solution, followed by 100 mL of ice water to quench the reaction. The mixture was vigorously stirred for 10 minutes and extracted with 300 mL of ethyl acetate. The organic phase was washed twice with water (100 mL each time), once with 100 mL of saturated sodium bicarbonate solution, and once with 300 mL of saturated aqueous sodium chloride solution. The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The product was separated by flash column chromatography (PE:EA=4:1, v:v) to obtain 6.35 g of compound 81a as a white solid in a yield of 70.1%. 1 H NMR (400MHz, CDCl3) δ8.15–8.08(m,2H),7.75(d,J=4.0Hz,1H),7.39–7.33(m,2H),6.68(d,J=4.0Hz,1H),2.43(s,3H).

[0290] Synthesis of compound 82a:

[0291] Compound 81a (4.0 g, 11.66 mmol), (S)-(-)-4-isopropyl-2-oxazolidinone (1.51 g, 11.66 mmol), potassium phosphate (4.95 g, 23.32 mmol), and Xantphos Pd-G3 (333 mg, 0.35 mmol) were dissolved in 115 mL of toluene and refluxed at 80°C under nitrogen for 4 h. The reaction was monitored by TLC. The reaction solution was diluted with 300 mL of ethyl acetate and transferred to a separatory funnel. The solution was washed twice with water (150 mL each) and once with 300 mL of saturated sodium chloride solution. The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The mixture was separated by flash column chromatography (PE:EA = 6:1, v:v) to afford 1.73 g of compound 82a as a white solid in a 34.2% yield. 1H NMR (400MHz, CDCl3) δ8.19–8.11(m,2H),7.66(d,J=4.1Hz,1H),7.41–7.35(m,2H),7.09(d,J=4.1Hz,1H),4.98(ddd,J=9.1,5.3,4.0Hz ,1H),4.49(t,J=8.9Hz,1H),4.36(dd,J=9.0,5.2Hz,1H),2.45(s,3H),1.30–1.25(m,1H),0.95(d,J=7.1Hz,3H),0.83(d,J=6.9Hz,3H).

[0292] Synthesis of compound 85a:

[0293] The synthesis of compound 85a was the same as compound I-17, to obtain 180 mg of compound 85a as a yellow solid, with a yield of 53%. 1 H NMR(500MHz, CDCl3)δ9.81(s,1H),8.60–8.53(m,3H),8.40(s,1H),7.85–7.79(m,3H),7.76–7.69(m,4 H),7.65(s,1H),7.65–7.55(m,3H),7.58–7.52(m,3H),7.53(s,1H),7.44(dd,J=8.0,5.0Hz,1H),7.37– 7.31(m,3H),7.27(dq,J=7.6,1.1Hz,3H),7.21(s,1H),6.88(s,1H),4.86(d,J=1.5Hz,1H),4.47(s,1H) ,4.26(s,1H),4.11(s,1H),2.41(t,J=1.0Hz,4H),2.28(s,1H),1.60(s,3H),1.03(s,3H),0.98(s,3H).

[0294] Synthesis of compound I-79:

[0295] Compound 85a (90 mg, 0.12 mmol) was dissolved in 2 mL of tetrahydrofuran and heated to 60°C under nitrogen. 2 mL of a 1 mol / L tetrabutylammonium fluoride solution in tetrahydrofuran was injected into the solution and allowed to react for 16 hours. After completion of the reaction, as monitored by TLC, heating was discontinued and the reaction solution was diluted with 45 mL of ethyl acetate and washed sequentially with saturated ammonium chloride solution, water, and saturated brine twice, 30 mL each time. The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The product was separated by flash column chromatography (DCM:MeOH = 50:1-25:1, v:v) to afford 50 mg of compound I-79 as a pale yellow solid in a 70% yield. 1H NMR (400MHz, DMSO-d6) δ11.20(s,1H),10.41(d,J=8.1Hz,1H),8.84(d,J=2.3 Hz,1H),8.59(dd,J=4.7,1.6Hz,1H),8.05(dt,J=8.1,2.0Hz,1H),7.78(d,J=8.4Hz,2H),7.69–7. 58(m,3H),7.55(d,J=8.3Hz,2H),7.49(dd,J=8.0,4.7Hz,1H),7.43(d,J=8.0Hz,2H),7.25(d,J=8. 1Hz,1H),7.01–6.87(m,2H),6.35(dd,J=3.6,2.0Hz,1H),5.15–5.01(m,1H),4.74(s,1H),4.46(t ,J=8.9Hz,1H),4.27(s,1H),1.96(d,J=32.0Hz,1H),1.47(d,J=7.1Hz,3H),1.17(t,J=7.3Hz,6H). 13 C NMR (126MHz, DMSO-d6) δ163.59,158.03,156.31,150.84,149.75,138.85,138.14,137.23,135.75,134.58,131.06,127.27 ,126.69,126.52,124.80,124.51,121.18,120.14,102.63,102.24,63.64,59.16,45.95,27.81,23.90,17.82,14.60,9.12. ESI-HRMS m / z:610.2534[M+H] + .

[0296] Example 50

[0297] Synthesis of compound I-80:

[0298] The synthesis method of compound I-80 was based on the synthesis of compound I-79 to obtain 90 mg of compound I-80 as a white solid with a yield of 76%. 1HNMR (400MHz, DMSO-d6) δ11.20(s,1H),10.27(s,1H),8.38(d,J=3.0Hz,1H),8.21(d,J=4.6Hz,1H),7.7 0(d,J=8.5Hz,2H),7.64–7.48(m,4H),7.47–7.39(m,3H),7.36(dd,J=8.4,4.5Hz,1H),7.25(d,J=8.1Hz, 1H),6.92(d,J=4.0Hz,1H),6.35(d,J=3.6Hz,1H),5.13–5.01(m,1H),4.83(s,2H),4.73(s,1H),4.46(t ,J=9.0Hz,1H),4.27(d,J=7.9Hz,1H),1.89(d,J=88.0Hz,1H),1.47(d,J=7.0Hz,3H),1.38–1.11(m,6H). 13 C NMR(101MHz,DMSO-d6)δ166.61,158.02,156.29,154.89,154.65,149.83,142.72,138.50,138.01,135.98,127.20,126.68 ,126.55,124.60,121.67,121.20,120.50,102.62,102.23,67.52,63.61,59.14,45.89,27.77,23.92,17.81,14.57,9.06. ESI-HRMS m / z:592.2663[M+H] + .

[0299] Example 51

[0300] Synthesis of compound 86a:

[0301] Compound 45a (600 mg, 2.46 mmol), compound 82a (1.07 g, 2.46 mmol), and DIPEA (2.2 g, 17.22 mmol) were dissolved in 3 mL of DMSO and heated to 110°C under reflux for 16 h under nitrogen. After completion of the reaction, as monitored by TLC, the reaction solution was diluted with 200 mL of ethyl acetate and transferred to a separatory funnel. The solution was washed with 100 mL of water (50 mL each) twice and 200 mL of saturated sodium chloride solution once. The organic phase was dried over anhydrous sodium sulfate and concentrated to dryness under reduced pressure. Chiral separation by flash column chromatography (DCM:EA = 40:1, v:v) afforded 254 mg of compound 86a as a yellow solid in a 16.0% yield. 1H NMR (300MHz, DMSO-d6) δ9.23(d,J=14.2Hz,2H),8.36(d,J=8.2Hz,2H),8.03(d,J=67.0Hz,5H),7.48(s,1H),7.28(d,J=18.8Hz,2H),6.66(d,J=4.1H z,1H),4.99(d,J=109.0Hz,2H),4.43(d,J=35.7Hz,2H),2.39(d,J=10.3Hz ,3H),2.22(s,1H),1.62(d,J=7.1Hz,3H),0.99–0.78(m,3H),0.38(s,3H).

[0302] Synthesis of compound I-81:

[0303] The synthesis method of compound I-81 was the same as that of compound I-79, to obtain 56 mg of compound I-81 as a light yellow solid with a yield of 80%. 1 H NMR (300MHz, DMSO-d6) δ11.27(s,1H),10.49(s,1H),9.08(s,2H),8.85(d,J=2. 2Hz,1H),8.60(dd,J=4.8,1.6Hz,1H),8.06(d,J=8.1Hz,1H),7.92–7.75(m,4H), 7.67(d,J=15.8Hz,1H),7.50(dd,J=8.0,4.8Hz,1H),7.04–6.86(m,3H),6.45(d d,J=3.7,1.9Hz,1H),5.18(t,J=7.2Hz,1H),4.70(s,1H),4.44(t,J=8.8Hz,1H), 4.33(dd,J=9.0,4.8Hz,1H), 2.41(td,J=7.0,3.8Hz,1H), 1.56(d,J=6.9Hz,3H), 0.91(d,J=7.0Hz,3H), 0.77(d,J=6.8Hz,3H). 13 C NMR (101MHz, DMSO-d6) δ170.91,163.74,157.82,156.23,154.97,154.83,150.93,149.79,140.13,137.68,134.67,130. 96,130.89,129.26,127.74,124.56,124.47,121.00,120.28,102.53,102.11,63.66,59.54,28.27,21.55,17.93,14.78. ESI-HRMS m / z:612.2434[M+H] +.

[0304] Example 52

[0305] Synthesis of compound I-82:

[0306] The synthesis method of compound I-82 was based on the synthesis of compound I-81 to obtain 58 mg of compound I-82 as a light yellow solid with a yield of 80%. 1 H NMR (300MHz, DMSO-d6) δ11.27(s,1H),10.34(s,1H),9.07(s,2H),8.39(dd,J=2.9,0.7Hz,1H),8.22(dd,J=4.5, 1.4Hz,1H),7.79(s,4H),7.45(ddd,J=8.5,2.9,1.4Hz,1H),7.37(ddd,J=8.5,4.6,0.8Hz,1H),7.01–6.83(m,2H) ,6.45(dd,J=3.7,1.9Hz,1H),5.28–5.09(m,1H),4.85(s,2H),4.70(s,1H),4.44(t,J=8.8Hz,1H),4.33(dd,J=8 .9,4.8Hz,1H),2.41(td,J=7.0,3.7Hz,1H),1.55(d,J=7.0Hz,3H),0.90(d,J=7.0Hz,3H),0.77(d,J=6.9Hz,3H). 13 C NMR(101MHz,DMSO-d6)δ170.96,166.84,157.81,156.22,154.99,154.83,154.64,149.91,142.71,139.33,138.44, 129.49,127.65,124.64,121.80,121.00,120.63,102.53,102.11,67.53,63.66,59.54,28.27,21.55,17.92,14.78. ESI-HRMS m / z:616.2385[M+H] + .

[0307] Example 53

[0308] 1. Determination of the inhibitory effect of compounds on mIDH1 activity

[0309] (1) IDH1-R132H / R132C

[0310] The compound to be tested was dissolved in DMSO to prepare a stock solution with a concentration of 10 mM, and then graded diluted with DMSO to the concentration to be tested.

[0311] Buffer formula: 150 mM NaCl, 20 mM Tris-HCl pH 7.5, 10 mM MgC12, 0.05% BSA.

[0312] Substrate mixture: a mixed solution of 20 μM NADPH and 5 mM α-ketoglutarate prepared in buffer.

[0313] Stop solution: a mixed solution of 36 μg / mL diaphorase and 30 mM resazurin prepared in buffer.

[0314] To a 384-well plate, add 1 μL of the test compound dissolved in DMSO and 39 μL of IDH1-R132H / IDH1-R132C enzyme diluted in buffer to a final concentration of 0.125 μg / mL. Incubate at room temperature for 1 hour. Add 10 μL of substrate mix to a 50 μL reaction volume to initiate the reaction and incubate at room temperature for 1.5 hours. Add 25 μL of stop solution and incubate at room temperature for 10 minutes. Fluorescence intensity (FI) is measured at excitation and emission wavelengths of 544 nm and 590 nm (Ex / Em = 544 / 590 nm). Greater inhibition of enzyme activity indicates a stronger fluorescence signal. The fluorescence signal of the wells without IDH-R132H / IDH-R132C was the strongest, which was recorded as FImax. The fluorescence signal of the solvent control wells (containing only buffer and DMSO) was the weakest, which was recorded as FImin. The fluorescence signal of the wells with compound addition was theoretically between FImin and FImax, which was recorded as FIx. The enzyme inhibition rate was calculated as follows: Enzyme inhibition rate (%) = [1-(FImax-FIx / FImax-FImin)] × 100%

[0315] All data were calculated and analyzed using GraphPad Prism software.

[0316] (II) IDH1-WT (wild-type IDH1)

[0317] The test compound was dissolved in DMSO to prepare a stock solution with a concentration of 10 mM, and then serially diluted with DMSO to the concentration to be tested.

[0318] Buffer formula: 150 mM NaCl, 20 mM Tris-HCl pH 7.5, 10 mM MgC12, 0.05% BSA.

[0319] Substrate mixture: a mixed solution of 0.35 mM isocitrate, 100 μg / mL diaphorase, and 200 μM resazurin prepared in buffer.

[0320] Add 1 μL of the test compound dissolved in DMSO and 39 μL of IDH1-WT enzyme diluted in buffer containing 62.5 μM NADP to a 384-well plate. + and 2.5 mM β-mercaptoethanol, for a final enzyme concentration of 0.02 μg / mL. This mixture was incubated at room temperature for 1 hour. Then, 10 μL of substrate mix was added to bring the reaction volume to 50 μL to initiate the reaction. The reaction was allowed to proceed at room temperature for 30 minutes and then stopped by adding 25 μL of 6% sodium dodecyl sulfate (SDS) solution. Fluorescence intensity was measured at excitation and emission wavelengths of 544 nm and 590 nm, respectively.

[0321] (3) Experimental results

[0322] Table 1. IC values ​​of compounds for mIDH1 and IDH1-WT inhibitory activities 50 (nM)

[0323] The results showed that most compounds had good activity against IDH1-R132H, better than IDH1-R132C, and had no obvious inhibition on normal IDH1 (IDH1-WT), indicating that the compounds may have good inhibitory activity against tumors with IDH1-R132H mutations and the inhibitory effect is better than that of tumors with IDH1-R132C mutations, and have good selectivity.

[0324] 2. Determination of the inhibitory effect of compounds on NAMPT activity

[0325] (1) Experimental methods

[0326] Enzyme reaction system: 25 μL, in which the final concentrations of various components are: 50 mM Tris-HCl (pH 7.5), 0.02% BSA, 12 mM MgCl2, 2 mM ATP, 0.4 mM PRPP, 2 mM DTT, 2 μg / mL NAMPT, 0.2 μM NAM, DMSO and serially diluted compounds.

[0327] First, 0.5 μL of compound solutions dissolved in DMSO of different concentrations were added to a 96-well plate, and then 20 μL of the above enzyme reaction mixture (enzyme reaction components other than the substrate) was added. After incubation at room temperature for 5 minutes, 4.5 μL of substrate nicotinamide (NAM) aqueous solution was added to start the reaction. After reacting at 37°C for 15 minutes, the enzyme reaction was terminated by heating at 95°C for 1 minute. After the reaction solution was cooled on ice, 10 μL of 20% acetophenone (dissolved in DMSO) and 2M KOH were added in sequence. After mixing on a vortex mixer, the mixture was incubated at 0°C for 2 minutes. 45 μL of 88% formic acid was added and the mixture was incubated at 37°C for 10 minutes. The fluorescence value at an excitation wavelength of 382 nm and an emission wavelength of 445 nm was measured using a microplate reader. The curve of fluorescence and inhibitor concentration was plotted to obtain the IC 50 value.

[0328] (2) Experimental results

[0329] Table 2. IC values ​​of compounds for NAMPT inhibition 50 (nM)

[0330] As can be seen from Tables 1 and 2, some compounds of the present invention have very good inhibitory activity against mIDH1 / NAMPT and selectivity for normal IDH1, especially compounds I-16, I-17, I-18, I-26, I-37, I-41, I-42, I-43, I-44, I-45, I-46, I-47, I-48, I-49, I-50, I-51, I-52, I-59, I-60, I-61, I-79, I-80, I-81, and I-82 have more prominent inhibitory activity against mIDH1 / NAMPT.

[0331] 3. Determination of the blood-brain barrier penetration ability of compounds

[0332] Experimental steps:

[0333] 1. Solution Preparation

[0334] 20% bovine brain phospholipid (PBL)-dodecane solution: Prepare a 20% bovine brain phospholipid (PBL)-dodecane solution by volume, sonicate until completely dissolved, and dilute the 20% bovine brain phospholipid-dodecane solution to 2% with dodecane.

[0335] Compound stock solution concentration: Weigh a certain amount of compound and prepare a stock solution with a concentration of 5 mg / mL or a molar concentration of 10 mM for later use.

[0336] Preparation of compound stock solution: 10 μL of compound stock solution was diluted 100-fold with 20% acetonitrile in pH 7.4 PBS to prepare a secondary stock solution (final concentration 50 μg / mL);

[0337] 2. Experimental Operation

[0338] (1) Drawing of the standard curve of the drug to be tested

[0339] Dilute the 5 mg / mL primary stock solution (positive control, negative control, and compound) by half to create standard solutions at concentrations of 1.67, 0.56, 0.18, 0.062, 0.021, 0.0068, and 0 mg / mL. Measure the peak areas of these solutions using HPLC. Create a standard curve using peak area versus compound concentration. Analyze whether the samples exhibit good linearity in the 0-1.67 mg / mL range.

[0340] (2) Determination of the ability of compounds to penetrate the blood-brain barrier

[0341] (a) Wetting: Add 50-100 μL of 70% ethanol to the wells of a 96-well filter plate. After 30 seconds, aspirate the plate and wash with 100 μL of PBS. Repeat twice, aspirating all PBS in the second wash.

[0342] (b) Coating: 4-5 μL of sonicated PBL-dodecane solution (2%) was added to the filter membrane of the receptor plate and 150 or 200 μL of pH 7.4 PBS buffer was added;

[0343] (c) Add 300 μL of test compound to the donor plate;

[0344] (d) The test compound diffuses from the donor wells through the lipid membrane into the acceptor wells. After 6 or 18 h of incubation, permeation occurs. The drug concentrations in the acceptor, donor, and reference wells are determined using HPLC. The effective permeability (Pe, cm / s) of the compound is calculated.

[0345] Pe=-(218.3 / t)*log[1-2CA(t) / CD(t0)]×10 -6 (cm / s)

[0346] Where CA(t) is the concentration of the drug solution in the receiving plate at time t (mg / L or molar concentration); CD(t0) is the initial concentration of the solution in the donor plate (mg / L or molar concentration); t is the incubation time (s).

[0347] Table 3. Blood-brain barrier penetration ability of compounds Note: a CNS + Indicates high penetration, CNS -Indicates low permeability.

[0348] This demonstrates that compound I-51 has the ability to effectively penetrate the blood-brain barrier. The inventors also investigated the other 51 compounds, which also possess the ability to penetrate the blood-brain barrier. However, FK866 does not, and AG-120 has poorer ability to penetrate the blood-brain barrier.

[0349] After comprehensively considering the compound's enzyme activity, selectivity, solubility and blood-brain barrier penetration ability, compound I-51 was selected for further cell experiments and in vivo activity evaluation.

[0350] 4. Determination of the Effect of Compounds on Cell Viability

[0351] U87 MG cells or U87 MG IDH1-R132H Cells were grown in T125 flasks. U87 MG cells were cultured in DMEM supplemented with 10% FBS and 1x mycin / streptomycin. IDH1-R132H The cells were cultured in DMEM containing 10% FBS, 1x penicillin / streptomycin, and 1 μL / mL puromycin. When the cells grew to 80%, they were digested and collected with trypsin to obtain U87 MG or U87 MG. IDH1-R132H A cell suspension (DMEM containing 10% FBS) was seeded into a 96-well white-bottom plate at a density of 5000 cells / well at 100 μL / well and incubated overnight at 37°C in 5% CO2. The next day, 100 μL of the test compound was added to each well in a serial dilution series to a final DMSO concentration of 0.2%. The 96-well plate was then placed in an incubator for 72 hours. 10 μL of CCK8 solution was then added to each well and the plate was placed in an incubator for 4 hours. The absorbance (OD) at 450 nm was measured using a microplate reader. The experimental results are as follows:

[0352] Table 4. Effects of compounds on cell viability Note: Positive-acting drugs AG-120 and IDH-305 are mIDH1 inhibitors, and positive-acting drug FK866 is a NAMPT inhibitor. The two-drug combination group consists of a 1:1 molar ratio of mIDH1 inhibitor to NAMPT inhibitor.

[0353] The results are shown in Table 4, indicating that compound I-51 can significantly inhibit the U87 MG IDH1-R132H The cell proliferation was significantly inhibited, and the inhibitory effect after 72 hours was better than that of the positive drug group and the drug combination group.

[0354] V. Determination of acute toxicity of compounds

[0355] Test animals: ICR mice (Jiangsu Huachuang Xinnuo Pharmaceutical Technology Co., Ltd.); 18-22 g; male; a total of 44 mice.

[0356] Test samples: Compound I-51, IDH-305, and FK866.

[0357] Compounds I-51, IDH-305, and FK866 were dissolved in DMSO, and HS (polyethylene glycol stearate) and PEG-400 were added as solubilizers in an amount equal to that of DMSO, and then diluted with normal saline. All three were injected through the tail vein at a dose of 400 mg / kg. After a single dose, the animal status was observed. Within 12 hours after administration, no abnormalities were observed in the animals in each group of mice. No animal deaths were observed within 24 hours of administration, and no animal deaths were observed after the 7th day of administration. No other obvious abnormalities were observed. On the 7th day, tissues such as the heart, liver, spleen, lung, and kidney were taken for H&E staining and pathological section observation (Figure 1), and the cell morphology of each tissue was good. No toxic reactions were observed in mice after intravenous administration of 400 mg / kg of the test drug, indicating that compound I-51 is safe.

[0358] 6. Pharmacokinetic parameters of compounds

[0359] Test animals: SD rats (180-200 g, male) were purchased from Jiangsu Huachuang Xinnuo Pharmaceutical Technology Co., Ltd.

[0360] Test sample: Compound I-51.

[0361] Experimental procedures: SD rats were intravenously injected with compound I-51 (1 mg / kg) and orally administered with compound I-51 (10 mg / kg). Orbital venous blood was collected at 2 min (no such time point for oral administration), 5 min, 15 min, 0.5 h, 1 h, 2 h, 4 h, 6 h, 8 h, 12 h, and 24 h after administration. Plasma was prepared and the compound concentration was detected by LC-MS / MS. The plasma clearance rate CL and elimination half-life T were investigated. 1 / 2 , peak time T max , peak concentration C max , area under the drug-dose curve AUC, apparent volume of distribution V ss , relative bioavailability F and other pharmacokinetic parameters.

[0362] Assay method: Compound I-51 was dissolved in 95% acetonitrile to prepare a series of working solutions. 10 μL of working solution (1, 2, 5, 10, 50, 100, 500, 1000, 5000, and 10,000 ng / mL) was added to 10 μL of blank SD rat plasma to create a total volume of 20 μL of 1×10,000 ng / mL compound I-51 calibration standard. 20 μL of standard and 20 μL of unknown sample (10 μL of unknown sample plasma and 10 μL of 95% acetonitrile) were added to 200 μL of acetonitrile to precipitate proteins. The samples were then vortexed for 30 seconds and centrifuged at 18,000 rpm at 4°C for 10 minutes. 10 μL of the supernatant was injected into the LC-MS / MS system for quantitative analysis.

[0363] The pharmacokinetic parameters (PK Parameters) of compound I-51 are shown in Table 5.

[0364] Table 5. Pharmacokinetic parameters of compound I-51 Note: iv: intravenous injection; po: oral administration; CL: apparent clearance; T 1 / 2 : Half-life; AUC 0-t : Area under the drug-time curve from the start of administration to the last point (24 hours); AUC 0-∞ T: the area under the drug-time curve from the start of drug administration to the theoretically extrapolated infinite time; max : peak time; C max : peak concentration; F: relative bioavailability.

[0365] VII. Determination of Anti-glioma Activity of Compounds

[0366] Test samples: Compound I-51, positive drugs (mIDH1 inhibitor IDH305, NAMPT inhibitor FK866).

[0367] Experimental method: Human brain glioma U87 MG in logarithmic growth phase was selected IDH1-R132H The cells were inoculated into 44 BALB / c nude mice at a depth of 3.0 mm, 1.0 mm in front of the midpoint of the anterior bregma and 2.5 mm to the right of the sagittal suture. The inoculation volume of cells per mouse was 1 × 10 544 tumor-bearing nude mice with good growth status and uniform tumor size were selected using a small animal imaging device and divided into 7 groups. Except for the model control group, which consisted of 8 tumor-bearing nude mice, the other 6 groups consisted of 6 mice each, namely (1) the model control group (administered with equal amounts of solvent, a mixed solvent of DMSO, HS, PEG-400, and normal saline in a volume ratio of 1:1:1:8); (2) the FK866 group (administered at a single dose of 30 mg / kg, ip, bid); (3) the IDH305 group (administered at a single dose of 300 mg / kg, po, bid); (4) the drug combination group (administered at a single dose of 30 mg / kg FK866 + 300 mg / kg IDH305, bid); (5) the low-dose group of compound I-51 (administered at a single dose of 10 mg / kg, iv, bid); and (6) the medium-dose group of compound I-51 (administered at a single dose of 20 mg / kg, iv, bid). (7) Compound I-51 high-dose group (single dose of 40 mg / kg, iv, bid). The nude mice were weighed every other day and euthanized 35 days after administration. The brain, heart, liver, spleen, lung, and kidney of the animals were surgically removed.

[0368] The experimental results showed that all animals in the model control group died around 13 days and had a significant decrease in body weight (Figure 2); the single drug groups (FK866 group and IDH305 group) were able to prolong the survival of the animals, with median survival times of 17 days and 23.5 days, respectively, and alleviate the weight loss of the animals; compared with the FK866 group, the drug combination group was able to significantly prolong the survival of the animals (26.5 days) (**P<0.01), and compared with the IDH305 group, there was no significant difference in the survival of the animals in the drug combination group; at different doses, compound I-51 was able to alleviate the weight loss of the animals and significantly prolong the survival of the animals. The survival times of mice in the low, medium, and high dose groups were 25.5, 30.5, and 34 days, respectively (**P<0.01), and tumor growth was inhibited in a concentration-dependent manner (Figures 3, 4, and 5). At doses of 20 mg / kg and 40 mg / kg, compound I-51 showed significantly better therapeutic effects than the drug combination group (**P<0.01), and H&E staining of pathological sections showed a significant reduction in tumor cells (Figure 6). IDH1-R132H The compound I-51 had a significant inhibitory effect on the growth of orthotopic transplanted tumors in nude mice, and the medium and high dose groups (20 mg / kg and 40 mg / kg) of compound I-51 had a significant inhibitory effect on the growth of human brain glioma cells U87 MG IDH1-R132H The anti-tumor activity of nude mice with orthotopic transplanted tumors was significantly stronger than that of the single positive drug group and the drug combination group.

Claims

1. mIDH1 / NAMPT dual-target inhibitor, characterized by: A compound selected from the group consisting of a compound having a structure as shown in Formula I or a pharmaceutically acceptable salt, racemate, stereoisomer, prodrug or solvate thereof: wherein Ring A is selected from substituted or unsubstituted 6-membered saturated or unsaturated heterocyclic ring, benzene ring and 5-membered saturated or unsaturated heterocyclic ring, or 5-6-membered saturated or unsaturated heterocyclic ring and 5-6-membered saturated or unsaturated heterocyclic ring; the substituent of the 5-6-membered saturated or unsaturated heterocyclic ring is at least one of halogen, amino, hydroxyl, carboxyl, cyano, olefin, and amide; the heteroatom in the 6-membered saturated or unsaturated heterocyclic ring, benzene ring and 5-membered saturated or unsaturated heterocyclic ring, 5-6-membered saturated or unsaturated heterocyclic ring and 5-6-membered saturated or unsaturated heterocyclic ring is selected from oxygen atoms and nitrogen atoms; X is selected from a linear or branched alkylene chain, a 1,4-piperazine subyl group or a 1,3-pyrrolidine subyl group; X may be interrupted one or more times by one, more or any combination of the following groups: -O-, -CONH-, NHCO-, -NHCONH-, -NH-, -S-, alkenylene, and the hydrogen of X may be replaced by one or more of the following groups: halogen, amino, hydroxyl, carboxyl, amide, cyano, olefin, alkyne, C1-C4 alkoxy, C1-C6 alkyl; L is selected from a covalent bond, a substituted or unsubstituted benzene ring, a 5-6 membered heterocyclic ring or a C1-C4 alkenyl group, a C1-C4 alkoxy group, or a C1-C4 amide group; the substituent of the benzene ring is at least one of a halogen, an amino group, a hydroxyl group, a carboxyl group, a cyano group, an olefin group, or an amide group; Ring B is selected from a substituted or unsubstituted benzene ring, a 5-6 membered saturated or unsaturated heterocyclic ring; the substituent of the benzene ring is at least one of halogen, amino, hydroxyl, carboxyl, cyano, olefin, and amide; the 5-6 membered saturated or unsaturated heterocyclic ring contains 1-2 heteroatoms, and the heteroatoms are selected from oxygen atoms and nitrogen atoms; R 4 Selected from hydrogen or C1-C4 alkyl; R 5 is selected from halogen, C1-C4 alkyl, substituted or unsubstituted phenyl, hydroxyl or tert-butoxy, the substituent of the benzene ring is at least one of halogen, amino, hydroxyl, carboxyl, cyano, olefin and amide; Or R4 and R5 are combined to form a benzene ring.

2. The mIDH1 / NAMPT dual-target inhibitor according to claim 1, characterized in that: In the compound of formula I, ring A is selected from: X is selected from: m=an integer from 0 to 8; n=an integer from 0 to 8; p=an integer from 0 to 8; L is selected from a benzene ring, a phenoxy group, a pyridine ring, a pyrimidine ring or a covalent bond; Ring B is selected from a benzene ring, a pyridine ring, a pyrimidine ring or an imidazole ring; R 4 is selected from hydrogen or methyl; R 5 is selected from fluorine, methyl, phenyl, hydroxyl or tert-butoxy; or R 4 With R 5 Combined to form a benzene ring.

3. The mIDH1 / NAMPT dual-target inhibitor according to claim 2, characterized in that: In the compound of formula I, ring A is selected from: X is selected from: m=0; n=0; L is selected from a benzene ring; Ring B is selected from a benzene ring or a pyridine ring; R 4 Selected from methyl; R 5 Selected from F, methyl, phenyl, hydroxy or tert-butoxy; or R 4 With R 5 Combined to form a benzene ring.

4. mIDH1 / NAMPT dual-target inhibitor, characterized by: A compound selected from the group consisting of a compound having a structure as shown in Formula II or a pharmaceutically acceptable salt, racemate, stereoisomer, prodrug or solvent compound thereof: Wherein, ring A is selected from: X is selected from: m=an integer from 0 to 8; n=an integer from 0 to 8; p=an integer from 0 to 8; L is selected from a benzene ring, a phenoxy group, a pyridine ring, a pyrimidine ring or a covalent bond; Ring B is selected from a benzene ring, a pyridine ring, a pyrimidine ring or an imidazole ring; R 1 Selected from sulfur, -NH-, -N(CH3)- or -CH-; R 2 is selected from nitrogen or -CH-; R 3 is selected from sulfur or -CH-; R 4 is selected from hydrogen or methyl; R 5 Selected from F, methyl, phenyl, hydroxy or tert-butoxy; or R 4 With R 5 Combined to form a benzene ring.

5. The mIDH1 / NAMPT dual-target inhibitor according to claim 4, characterized in that: In the compound shown in formula II, ring A is selected from: X is selected from: m=an integer from 0 to 8; n=an integer from 0 to 8; p=an integer from 0 to 8; L is selected from a benzene ring; Ring B is selected from a benzene ring, a pyridine ring or a pyrimidine ring; R 1 Selected from sulfur, R 2 Selected from -CH-, R 3 is selected from -CH-; or R 1 Selected from -CH-, R 2 Selected from -CH-, R 3 Selected from S; or R 1 Selected from -NH-, R 2 Selected from -CH-, R 3 Selected from -CH-; R 4 Selected from methyl; R 5 Selected from methyl.

6. The mIDH1 / NAMPT dual-target inhibitor according to claim 4, characterized in that: In the compound shown in Formula II, Ring A is selected from: X is selected from: n＝0； L is selected from a benzene ring; Ring B is selected from a benzene ring, a pyridine ring or a pyrimidine ring; R 1 Selected from -NH-, R 2 Selected from -CH-, R 3 Selected from -CH-; R 4 Selected from methyl; R 5 Selected from methyl.

7. mIDH1 / NAMPT dual-target inhibitor, characterized by: A compound selected from the group consisting of the following structures or pharmaceutically acceptable salts, racemates, stereoisomers, prodrugs or solvates thereof:

8. A pharmaceutical composition, characterized in that: Contains the mIDH1 / NAMPT dual-target inhibitor according to any one of claims 1 to 7 and a pharmaceutically acceptable carrier.

9. Use of the mIDH1 / NAMPT dual-target inhibitor according to any one of claims 1 to 7 in the preparation of drugs for diseases mediated by mIDH1 and / or NAMPT.

10. Use of the mIDH1 / NAMPT dual-target inhibitor according to any one of claims 1-7 in the preparation of a drug for treating cancer or tumor-related diseases; the cancer or tumor-related diseases include multiple myeloma, leukemia, breast cancer, prostate cancer, lung cancer, liver cancer, gastric cancer, bone cancer, brain cancer, head and neck cancer, intestinal cancer, pancreatic cancer, bladder cancer, testicular cancer, ovarian cancer, and endometrial cancer.

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