Aza-four-membered ring connected pyrazolopyridine compound or pharmaceutically acceptable salt thereof and application of aza-four-membered ring connected pyrazolopyridine compound or pharmaceutically acceptable salt thereof

Abstract

The invention discloses an aza-four-membered ring connected pyrazolopyridine compound or pharmaceutically acceptable salt and application thereof, and the compound with the structure can be used as a novel high-selectivity PDE10A inhibitor for treating peripheral tissue diseases such as cardiac hypertrophy and cardiac dysfunction (heart failure), diabetes, hypertension, renal function impairment and renal fibrosis. Lung inflammation and pulmonary fibrosis, immune regulation disorder and cancers (colon cancer, ovarian cancer and lung cancer). Through strategies in two aspects, the inhibition effect of the compound on a PDE10A high-selectivity inhibitor is reduced while the inhibition effect of the compound on a central nervous system is reduced. Firstly, in the aspect of molecular structure modification, the structure of the pyrazolopyridine compound connected with the aza-four-membered ring is optimized, for example, the polar surface area of molecules is increased by increasing the proportion of the number of N, O and S atoms in the molecular structure, and the fat solubility is reduced; and hydrophilic groups are introduced to increase water solubility and the like, so that the permeability of the compound to the blood brain barrier BBB is reduced. And 2, targeted drug delivery of lung tissues is realized by means of a pharmaceutical preparation.

Description

Technical Field

[0001] The invention relates to a pyrazolopyridine compound, in particular to a pyrazolopyridine compound connected with a nitrogen tetracyclic ring and application thereof, belonging to the technical field of pyrazolopyridine compounds. Background Art

[0002] As second messengers, cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP) are important regulators of intracellular signal transduction and are widely involved in physiological processes such as cell proliferation and apoptosis, vascular and airway smooth muscle relaxation, immune / inflammatory response, visual transduction, secretion and bone development. Cyclic nucleotide phosphodiesterases (PDEs) are a super enzyme family that specifically hydrolyzes cAMP and cGMP in human cells and determine the circulating concentrations of these two second messengers in cells. Among them, PDE4 / 7 / 8 specifically hydrolyzes cAMP, PDE5 / 6 / 9 specifically hydrolyzes cGMP, and PDE1 / 2 / 3 / 10 / 11 can hydrolyze both cAMP and cGMP at the same time, but the degree of preference for hydrolysis is different.

[0003] Prior art has shown that phosphodiesterase 10A (PDE10A) is abundantly expressed in the central striatum of the human brain and is involved in the regulation of information processing in the cortex-basal ganglia circuit, suggesting that it may play a role in the pathogenesis of neuropsychiatric diseases.

[0004] At present, the vast majority of small molecule drugs developed for PDE10A targets are concentrated in the treatment of central nervous system diseases (schizophrenia, Huntington's disease), and several representative drugs such as MP-10, AMG-579MK8189 and TAK-063 have entered clinical phase I / II trials, which requires that such drug molecules must have good blood-brain barrier BBB permeability to meet their accumulation in brain tissue and maintain a certain drug concentration. However, these PDE10A inhibitors are not suitable for the treatment of peripheral tissue diseases, mainly because their good BBB permeability can easily lead to a strong inhibitory effect on PDE10A highly expressed in the brain, significantly increasing the occurrence of clinical adverse events.

[0005] Therefore, it is extremely challenging to achieve that PDE10A inhibitors exert therapeutic effects on peripheral tissue diseases without producing obvious inhibitory effects on the central nervous system. To this end, a class of nitrogen-containing four-membered ring-linked pyrazolopyridine compounds and their applications are designed to solve the above problems. Summary of the invention

[0006] The main purpose of the present invention is to provide a class of pyrazolopyridine compounds connected by nitrogen four-membered rings and applications thereof.

[0007] The purpose of the present invention can be achieved by adopting the same technical solution as the technical solution in the claims, which will not be repeated again.

[0008] Beneficial technical effects of the present invention:

[0009] The present invention provides a class of pyrazolopyridine compounds connected by nitrogen-containing four-membered rings and their applications, aiming to find that such structural compounds can be used as new highly selective PDE10A inhibitors for the treatment of peripheral tissue diseases including: myocardial hypertrophy and cardiac dysfunction (heart failure), diabetes, hypertension, renal damage and renal fibrosis, lung inflammation and pulmonary fibrosis, immune regulation disorders, cancer (colon cancer, ovarian cancer and lung cancer), etc. Through two-way strategies, such compounds can be highly selective inhibitors of PDE10A while reducing their inhibitory effects on the central nervous system. 1. Molecular structure modification: By optimizing the structure of pyrazolopyridine compounds connected by nitrogen-containing four-membered rings, such as: increasing the ratio of the number of N, O, and S atoms in the molecular structure to increase the polar surface area of ​​the molecule and reduce fat solubility; introducing hydrophilic groups to increase water solubility, etc., the permeability of the compound to the BBB is reduced. Second, targeted drug delivery to lung tissue can be achieved through drug preparations (e.g., inhalable dry powder solid particle drug-loaded microspheres), thereby reducing the first-pass effect in the body caused by oral administration, lowering the dosage and systemic exposure of drug molecules, thereby reducing the accumulation of such compounds in the brain and the inhibitory effect on the central nervous system, and significantly reducing the potential risk of adverse reactions. BRIEF DESCRIPTION OF THE DRAWINGS

[0010] Figure 1 The heart weight ratio and heart-to-tibia ratio of a preferred embodiment B16 of a class of nitrogen tetracyclic ring-linked pyrazolopyridine compounds and their use according to the present invention after drug administration intervention are shown in Figure 1. *P<0.05, **P<0.01, ***p<0.001, ****p<0.0001, ns means no statistical difference, n=6.

[0011] Figure 2 The following is a statistical diagram of the changes in the expression of atrial natriuretic peptide (ANP) and β-myosin heavy chain (β-MHC) mRNA in myocardial tissue after administration of a preferred embodiment B16 of a class of nitrogen tetracyclic ring-linked pyrazolopyridine compounds and their applications according to the present invention. *P<0.05, **P<0.01, ***p<0.001, ****p<0.0001, ns means no statistical difference, n=6.

[0012] Figure 3The left ventricular end-systolic diameter and left ventricular end-diastolic diameter statistical graphs after drug administration intervention according to a preferred embodiment B16 of a class of nitrogen-four-membered ring-connected pyrazolopyridine compounds and their uses of the present invention. *P<0.05, **P<0.01, ***p<0.001, ****p<0.0001, ns means no statistical difference, n=6.

[0013] Figure 4 The left ventricular ejection fraction and left ventricular fractional shortening statistical graphs after administration of a preferred embodiment B16 of a class of nitrogen tetracyclic ring-linked pyrazolopyridine compounds and their use according to the present invention. *P<0.05, **P<0.01, ***p<0.001, ****p<0.0001, ns means no statistical difference, n=6.

[0014] Figure 5 This is a preferred embodiment B16 of a class of nitrogen-four-membered ring-linked pyrazolopyridine compounds and their uses according to the present invention, and staining of myocardial slices after drug administration intervention, wherein H&E represents hematoxylin staining; WGA represents wheat germ agglutinin staining; picrosirius red represents picrosirius red staining, ISO represents isoproterenol, and propranolol represents propranolol.

[0015] Figure 6 A preferred embodiment B11 of a class of nitrogen tetracyclic ring-linked pyrazolopyridine compounds and their use according to the present invention improves lung function (WBP method) in mice with pulmonary fibrosis after administration intervention. (A) Enhanced pause (Penh); (B) End expiratory pause (EEP); (C) Relaxation time (RT). Compared with the normal control group, # P<0.05, ## P<0.01; compared with the model group, *P<0.05, **P<0.01, ns means no statistical difference, n=6.

[0016] Figure 7This is a preferred embodiment B11 of a class of nitrogen-containing four-membered ring-connected pyrazolopyridine compounds and their applications according to the present invention. After drug administration, the lung function of mice with pulmonary fibrosis was improved (PFT method). (A) Airway resistance (Resistance Index, RI); (B) Dynamic compliance (Dynamic compliance, Cdyn); (C) Total lung capacity (Total lung capacity, TLC); (D) Inspiratory capacity (Inspiratory capacity, IC); (E) Forced vital capacity (Forced vital capacity, FVC); (F) Forced expiratory volume 100 in the first second (Forced expiratory volume 100, FEV100); Compared with the normal control group, compared with the normal control group, # P<0.05, ## P<0.01; compared with the model group, *P<0.05, **P<0.01, ns means no statistical difference, n=6.

[0017] Figure 8 A preferred embodiment B11 of a class of nitrogen-containing four-membered ring-connected pyrazolopyridine compounds and their use according to the present invention improves and reduces lung tissue lesions in mice with bleomycin-induced pulmonary fibrosis. (A) Representative images of HE and Masson; (B) H&E score; (C) Masson score. Scale bar: 100 μm; Compared with the normal control group, # P<0.05, ## P<0.01; compared with the model group, *P<0.05, **P<0.01, ns means no statistical difference, n=6.

[0018] Fig. 9 A schematic diagram of the preparation of a compound B11 nanosuspension and a dry powder inhaler thereof according to a preferred embodiment of a class of nitrogen-four-membered ring-connected pyrazolopyridine compounds and their applications of the present invention;

[0019] Fig.10 (A) Physical appearance and nanoparticle size distribution of a nanosuspension of compound B11 according to a preferred embodiment of a class of nitrogen-containing four-membered ring-connected pyrazolopyridine compounds and their uses of the present invention; (B) Transmission electron microscopy result;

[0020] Fig.11 (A) morphology and (B) particle size distribution of a drug-free dry powder according to a preferred embodiment of a class of nitrogen-containing four-membered ring-connected pyrazolopyridine compounds and their applications of the present invention; (C) morphology and (D) particle size distribution of a drug-loaded B11 dry powder;

[0021] Fig.12The in vitro atomization particle size distribution diagram of the drug-loaded dry powder according to a preferred embodiment B11 of a class of nitrogen-four-membered ring-connected pyrazolopyridine compounds and their use in the present invention. MMAD: median particle size, EF: emission fraction, FPF: fine particle fraction.

[0022] Fig.13 The invention relates to the synthesis of a class of pyrazolopyridine compounds A1-A12 connected by nitrogen four-membered rings according to the present invention.

[0023] Fig.14 The invention relates to the synthesis of intermediates M5 to M11 of pyrazolopyridine compounds connected by nitrogen four-membered rings.

[0024] Fig.15 The invention relates to the synthesis of a class of pyrazolopyridine compounds A13-A20 connected by nitrogen four-membered rings according to the present invention.

[0025] Fig.16 The invention relates to the synthesis of a class of pyrazolopyridine compounds B1 to B6 connected by nitrogen four-membered rings according to the present invention.

[0026] Fig.17 The invention relates to the synthesis of a class of pyrazolopyridine compounds B7-B22 connected by nitrogen four-membered rings according to the present invention.

[0027] Fig.18 The invention relates to the synthesis of a class of nitrogen-containing four-membered ring-linked pyrazolopyridine compounds B23 according to the present invention.

[0028] Fig.19 The invention relates to the synthesis of a class of pyrazolopyridine compounds B24 to B26 connected by nitrogen four-membered rings according to the present invention.

[0029] Fig. 20 The invention relates to the synthesis of a class of nitrogen-containing four-membered ring-linked pyrazolopyridine compounds B27 according to the present invention.

[0030] Fig.21 The invention relates to the synthesis of a class of pyrazolopyridine compounds C1-C2 connected by nitrogen four-membered rings according to the present invention.

[0031] Fig. 22 The invention relates to the synthesis of a class of pyrazolopyridine compounds C3 connected by nitrogen four-membered rings according to the present invention.

[0032] Fig.23 The invention relates to the synthesis of a class of pyrazolopyridine compounds C4-C8 connected by nitrogen four-membered rings.

[0033] Fig.24 The invention relates to the synthesis of a class of pyrazolopyridine compounds C9-C10 connected by nitrogen four-membered rings according to the present invention. DETAILED DESCRIPTION

[0034] In order to make the technical solution of the present invention more clear and specific to those skilled in the art, the present invention is further described in detail below in conjunction with embodiments and drawings, but the implementation manner of the present invention is not limited thereto.

[0035] In order to better illustrate the purpose, technical solutions and advantages of the present invention, the present invention will be further described below in conjunction with specific embodiments.

[0036] (1) Representative compound structures

[0037]

[0038]

[0039]

[0040]

[0041] (2) Synthesis of representative compound structures

[0042] In this example, the synthetic routes of compounds A1 to A12 (see Fig.13 ):

[0043] The synthesis process of intermediate M1 is as follows: In a 250 mL eggplant-shaped bottle, compound 1-Boc-azetidine-3-carboxylic acid (10 g, 49.7 mmol), cycloisopropyl malonate (10.74 g, 74.5 mmol) and 4-dimethylaminopyridine (9.10 g, 1.5 mmol) are dissolved in dichloromethane (100 mL). Under argon protection, a dichloromethane (60 mL) solution of N, N'-carbonyldiimidazole (9.67 g, 59.6 mmol) is slowly added dropwise to the above system at 0 ° C. The reaction mixture naturally rises to room temperature and is stirred overnight. The reaction is monitored by TLC. After the reaction is completed, a saturated citric acid aqueous solution is slowly added to the reaction system to adjust the pH to 5-6, saturated brine (60 mL) is added, the organic phase is separated, and the aqueous phase is extracted twice with dichloromethane (30 mL). The organic phase is separated, dried over anhydrous sodium sulfate, and the filtrate is filtered and concentrated by vacuum distillation to obtain a yellow oily compound. The compound was dissolved in anhydrous ethanol (60 mL), heated to 80°C with stirring and reacted overnight, and the reaction was monitored by TLC. After the reaction was completed, the reaction mixture was concentrated by distillation under reduced pressure, and the obtained crude product was purified by silica gel column chromatography to obtain a light yellow oily intermediate compound M1 (10.28 g, yield 76%). 1H NMR (400MHz, CDCl3) δ4.20 (q, J = 7.2Hz, 2H), 4.13–4.01 (m, 4H), 3.67–3.56 (m, 1H), 3.47 (s, 2H), 1.43 (s, 9H), 1.29 (t, J = 7.2Hz, 3H).

[0044] The synthesis process of intermediate M2a is as follows: In a 125mL eggplant-shaped bottle, intermediate M1 (3.02g, 11.13mmol) is dissolved in acetonitrile (60mL), and 2-aminopyridine (3.15g, 33.39mmol) and carbon tetrabromide (7.38g, 22.26mmol) are added. The reaction mixture is stirred overnight at 80°C, and the reaction is monitored by TLC. After the reaction is completed, the crude product is concentrated by vacuum distillation, to which a saturated aqueous solution of citric acid (30mL) is added, and ethyl acetate is extracted twice (30mLx 2), the organic phase is collected, washed with saturated brine, and purified by silica gel column chromatography after vacuum distillation to obtain a yellow oily intermediate compound M2a (2.58g, yield 61%). 1 H NMR (400MHz, CDCl3) δ9.33(d,J=6.8Hz,1H),7.72(d,J=8.8Hz,1H),7.43(t,J=6.8Hz ,1H),7.03(t,J=6.8Hz,1H),4.50–4.27(m,7H),1.46(s,9H),1.44(t,J=6.8Hz,3H).

[0045] The synthesis process of intermediate M3a is as follows: In a 125 mL eggplant-shaped bottle, intermediate M2a (2.58 g, 7.47 mmol) was dissolved in dichloromethane (40 mL), and trifluoroacetic acid (4 mL) was added. The reaction mixture was stirred overnight at room temperature, and the reaction was monitored by TLC. After the reaction was completed, the intermediate compound M3a (1.76 g, yield 96%) was obtained by vacuum distillation and concentration. 1 H NMR (400MHz, CDCl3) δ9.33(d,J=7.2Hz,1H),7.72(d,J=8.8Hz,1H),7.44(t,J=6.8Hz,1H ),7.01(t,J=6.8,1H),4.43–4.32(m,5H),4.46(q,J=7.2Hz,2H),1.47(t,J=7.2Hz,3H).

[0046] The synthesis process of compound A1 is as follows: in a 50 mL eggplant-shaped bottle, the intermediate M3a (100 mg, 0.41 mmol) and 2-chloroquinoline (80 mg, 0.49 mmol) are dissolved in anhydrous DMF (10 mL), and cesium carbonate (267 mg, 0.82 mmol) is added. The reaction mixture is heated to 110°C for overnight reaction, and the reaction is monitored by TLC. After the reaction is completed, cesium carbonate is filtered off, the filtrate is concentrated by distillation under reduced pressure, and the crude product is purified by silica gel column chromatography to obtain white solid compound A1 (72 mg, yield 62%). 1 H NMR (500MHz, CDCl3) δ9.34(d,J=7.0Hz,1H),7.86(d,J=9.0Hz,1H),7.74(d,J=8.5Hz,1H),7. 68(d,J=9.0Hz,1H),7.58(d,J=8.0Hz,1H),7.52(t,J=7.5Hz,1H),7.39(t,J=7.5Hz,1H),7.20 (t,J=7.5Hz,1H),7.00(t,J=7.0Hz,1H),6.66(d,J=9.0Hz,1H),4.69(dd,J=14.5,7.0Hz,1H), 4.63(t,J=7.5Hz,4H),4.45(q,J=7.0Hz,2H),1.47(t,J=7.0Hz,3H),HRMS(ESI-TOF)m / z[M+H] + Calculate for C 22 H 20 N4O2393.1659, found 373.1668.

[0047] In this example, the synthetic routes of intermediates M5 to M11 (see Fig.14 )as follows:

[0048] The synthesis process of intermediate M5 is as follows: Add compound 2-quinolinone-4-carboxylic acid (9.45 g, 50.0 mmol) to a 125 mL eggplant-shaped bottle, and carefully add phosphorus oxychloride (30 mL). Heat the reaction mixture to 110° C. and stir to react overnight, and monitor the reaction by TLC. After the reaction is completed, remove phosphorus oxychloride by distillation under reduced pressure to obtain yellow oily intermediate compound M5 (yield 100%).

[0049] The synthesis process of intermediate M6 is as follows: dry methanol (40 mL) is slowly added dropwise to the cooled intermediate M5 (50.0 mmol) at 0°C, and then the reaction mixture is stirred at room temperature for 2 hours, and the reaction is monitored by TLC. After the reaction is completed, saturated sodium bicarbonate solution is added to the reaction mixture at 0°C to adjust the pH of the system to 9-10, the aqueous phase is extracted twice with ethyl acetate, the organic phase is dried over anhydrous sodium sulfate, the filtered filtrate is concentrated by vacuum distillation, and the obtained crude product is purified by silica gel column chromatography to obtain a white solid intermediate compound M6 (5.87 g, 2-step yield 53%). 1 HNMR (400MHz, CDCl3) δ8.73 (d, J = 8.8 Hz, 1H), 8.08 (d, J = 8.4 Hz, 1H), 7.91 (s, 1H), 7.79 (td, J = 6.8, 1.2 Hz, 1H), 7.66 (td, J = 6.8, 1.2 Hz, 1H), 4.05 (s, 3H).

[0050] The synthesis process of the intermediate 2-chloro-N-methylquinoline-4-carboxamide M7 is as follows: the cooled intermediate M5 (50.0 mmol) is dissolved in dry dichloromethane (40 mL), and methylamine hydrochloride (4.05 g, 60.0 mmol) and triethylamine (15.2 g, 150.0 mmol) are added. The reaction mixture is stirred at room temperature for 3 hours, and the reaction is monitored by TLC. After the reaction is completed, a saturated sodium bicarbonate solution is added to the reaction mixture at 0°C to adjust the pH to 9-10, and dichloromethane is extracted twice. The organic phase is dried over anhydrous sodium sulfate, and the filtrate is filtered and concentrated by vacuum distillation. The obtained crude product is purified by silica gel column chromatography to obtain a white solid intermediate compound M7 (8.62 g, 2-step yield 78%). 1 HNMR(500MHz,DMSO-d6δ8.84(d,J=4.0Hz,1H),8.15(d,J=8.0Hz,1H),8.02(d,J=8.5Hz,1H), 7.88(td,J=7.0,1.0Hz,1H), 7.71(td,J=7.0,1.0Hz,1H), 7.65(s,1H), 2.87(d,J=4.5Hz,3H).

[0051] The synthesis process of intermediate M8 is as follows: using intermediate M5 (1.0 mmol) and 2,2-difluoroethylamine (97.2 mg, 1.2 mmol) as raw materials, referring to the synthesis method of intermediate M7, an off-white solid intermediate compound M8 (60 mg, yield 48%) was obtained.

[0052] 1H NMR (400MHz, CDCl3) δ8.14(d,J=8.4Hz,1H),8.05(d,J=8.4Hz,1H),7.79(t,J=7.2Hz,1H),7.63 (t,J=7.2Hz,1H),7.46(s,1H),6.41(brs,1H),6.07(tt,J=55.6,3.6Hz,1H),4.01–3.86(m,2H).

[0053] The synthesis process of intermediate M9 is as follows: using intermediate M5 (1.0 mmol) and N-methylpiperazine (120 mg, 1.2 mmol) as raw materials, referring to the synthesis method of intermediate M7, a light yellow solid intermediate compound M9 (60 mg, yield 48%) was obtained. 1 HNMR(400MHz, CDCl3)δ8.06(d,J=8.4Hz,1H),7.83–7.75(m,2H),7.61(t,J=7.6Hz,1H),7.31(s ,1H),4.04–3.84(m,2H),3.77–3.66(m,1H),3.29–3.11(m,3H),2.64–2.51(m,2H),2.33(s,3H).

[0054] The synthesis process of intermediate M10 is as follows: using intermediate M5 (1.0 mmol) and tert-butyl 4,7-diazaspiro[2.5]octane-4-carboxylate (254 mg, 1.2 mmol) as raw materials, referring to the synthesis method of intermediate M7, a light yellow solid intermediate compound M10 (290 mg, yield 72%) was obtained. 1 H NMR(400MHz, CDCl3)δ8.06(d,J=8.0Hz,1H),7.82–7.73(m,2H),7.60(t,J=7.6Hz,1H),7.31(s,1H),4.17–4.06(m,0.5H),3.92 –3.70(m,2H),3.68–3.51(m,1H),3.43–3.17(m,2H),3.05–2.95(m,0.5H),1.49(s,9H),1.21–0.86(m,3H),0.70–0.36(m,1H).

[0055] The synthesis process of intermediate M11 is as follows: using intermediate M5 (1.0 mmol) and N,N-diethylethylenediamine (139 mg, 1.2 mmol) as raw materials, referring to the synthesis method of intermediate M7, a light yellow solid intermediate compound M11 (250 mg, yield 82%) was obtained. 1HNMR (400MHz, CDCl3) δ8.24(d,J=8.4Hz,1H),8.05(d,J=8.0Hz,1H),7.77(td,J=7.2,1.2Hz,1H),7.61(td,J=7.2,1.2Hz,1H), 7.45(s,1H),6.90(brs,1H),3.60(dd,J=11.2,5.2Hz,2H),2.72(t,J=6.0Hz,2H),2.59(q,J=7.2Hz,4H),1.03(t,J=7.2Hz,6H).

[0056] The synthesis process of compound A2 is as follows: using intermediate M3a (100 mg, 0.41 mmol) and intermediate M6 (108 mg, 0.49 mmol) as raw materials and referring to the synthesis method of compound A1, a white solid compound A2 (121 mg, yield 69%) was obtained. 1 HNMR(400MHz, CDCl3) δ9.34(d,J=6.8Hz,1H),8.45(d,J=8.0Hz,1H),7.77(d ,J=8.8Hz,1H),7.69(d,J=8.8Hz,1H),7.56(t,J=7.6Hz,1H),7.41(t,J=8.0H z,1H),7.32–7.24(m,2H),7.02(t,J=6.8Hz,1H),4.79–4.60(m,5H),4.47(q, J=7.2Hz,2H),4.01(s,3H),1.49(t,J=7.2Hz,3H).HRMS(ESI-TOF)m / z:[M+H] + Calculate for C 24 H 22 N4O4431.1714, found431.1715.

[0057] The synthesis process of compound A3 is as follows: using intermediate M3a (1.76 g, 7.18 mmol) and intermediate M7 (1.58 g, 7.18 mmol) as raw materials and referring to the synthesis method of compound A1, a light yellow solid compound A3 (1.97 g, yield 64%) was obtained. 1HNMR(400MHz, CDCl3) δ9.34(d,J=6.8Hz,1H),7.88(d,J=8.4Hz,1H),7.75(brs,1 H),7.69(d,J=8.8Hz,1H),7.52(t,J=8.0Hz,1H),7.42(t,J=8.0Hz,1H),7.20(t,J =7.6Hz,1H),7.03(t,J=6.8Hz,1H),6.59(s,1H),4.70–4.50(m,5H),4.46(q,J=7. 2Hz,2H),3.11(d,J=5.2Hz,3H),1.49(t,J=7.2Hz,3H).HRMS(ESI-TOF)m / z:[M+H] + Calculate for C 24 H 23 N5O3430.1874,found430.1888.

[0058] The synthesis process of intermediate M4 is as follows: in a 125mL eggplant-shaped bottle, intermediate A3a (1.97g, 4.59mmol) is dissolved in methanol / water (30mL / 10mL), sodium hydroxide (550mg, 13.76mmol) is added, the reaction mixture is heated at 65°C and stirred for 3 hours, and the reaction is monitored by TLC. After the reaction is completed, methanol is removed by distillation under reduced pressure to obtain an aqueous solution of the crude product, the pH of the system is adjusted to 4-5 with 5M hydrochloric acid, a solid is precipitated, and the filter cake is washed with water under reduced pressure and dried in air to obtain a white solid intermediate compound M4 (1.51g, yield 82%). 1 H NMR (400MHz, CH3OD) δ9.33(d,J=6.8Hz,1H),7.88(d,J=8.0Hz,1H),7.75(s,1H),7.68(d,J=9.2Hz,1H),7.52(t,J=7.6Hz,1H),7.46– 7.39(m,1H),7.20(t,J=7.6Hz,1H),7.03(t,J=6.8Hz,1H),6.58(s,1H),4.63–4.61(m,1H),4.56–4.51(m,4H),3.13(d,J=4.8Hz,3H).

[0059] The synthesis process of compound A4 is as follows: in a 50 mL eggplant-shaped bottle, the intermediate M4 (100 mg, 0.25 mmol) is dissolved in anhydrous N, N-dimethylformamide (10 mL), and compound 2,2-difluoroethanol (41 mg, 0.50 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (58 mg, 0.30 mmol) and 4-dimethylaminopyridine (61 mg, 0.50 mmol) are added in sequence, and the reaction mixture is stirred at room temperature overnight, and the reaction is monitored by TLC. After the reaction is completed, saturated brine is added, ethyl acetate is extracted, and the organic phase is distilled under reduced pressure to obtain a crude product, which is purified by silica gel column chromatography to obtain a light yellow solid compound A4 (87 mg, yield 75%). 1 H NMR (500MHz, DMSO-d6) δ9.23(d,J=6.5Hz,1H),8.64(d,J=2.0Hz,1H),7.89(d,J=7.5Hz,1H),7.80(d,J=9.0Hz,1H),7.68–7.59(m,2H),7.56(d,J=6.5 Hz,1H),7.33–7.19(m,2H),6.82(s,1H),6.53(t,J=54.5Hz,1H),4.74–4.6 2(m,3H),4.56(s,2H),4.48(s,2H),2.84(s,3H).HRMS(ESI-TOF)m / z:[M+H] + Calculate for C 24 H 21 F2N5O3466.1685,found466.1690.

[0060] The synthesis process of compound A5 is as follows: using intermediate M4 (100 mg, 0.25 mmol) and isopropanol (30 mg, 0.5 mmol) as raw materials, referring to the synthesis method of compound A4, a light yellow compound A5 (66 mg, yield 60%) was obtained. 1H NMR (500MHz, CDCl3) δ9.34(d,J=7.0Hz,1H),7.80(d,J=8.0Hz,1H),7.68(d,J=9.0Hz,1 H),7.63(d,J=8.0Hz,1H),7.48(t,J=7.5Hz,1H),7.41(t,J=7.5Hz,1H),7.13(t,J=7.5 Hz,1H),7.02(t,J=7.0Hz,1H),6.39(s,1H),5.41–5.30(m,1H),4.64–4.55(m,1H),4.5 3–4.40(m,4H),3.11(d,J=4.5Hz,3H),1.46(d,J=6.0Hz,6H).HRMS(ESI-TOF)m / z:[M+H] + calcdfor C 25 H 25 N5O3444.2030,found 444.2032.

[0061] The synthesis process of compound A6 is as follows: using intermediate M4 (100 mg, 0.25 mmol) and oxetan-3-ol (37 mg, 0.5 mmol) as raw materials, referring to the synthesis method of compound A4, a light yellow compound A6 (77 mg, yield 68%) was obtained. 1 HNMR(400MHz, CDCl3)δ9.28(d,J=6.8Hz,1H),7.84(d,J=8.0Hz,1H),7.72(d,J=9.2Hz,1H), 7.65(d,J=8.4Hz,1H),7.57–7.40(m,2H),7.17(t,J=7.6Hz,1H),7.06(t,J=6.8Hz,1H),6.78 (q,J=4.0Hz,1H),6.49(s,1H),5.81–5.69(m,1H),5.06(t,J=7.2Hz,2H),4.84(t,J=7.2Hz, 2H),4.72–4.61(m,1H),4.59–4.48(m,4H),3.10(d,J=4.8Hz,3H).HRMS(ESI-TOF)m / z:[M+H] + Calculate for C 25 H 23 N5O4458.1823,found 458.1826.

[0062] The synthesis process of compound A7 is as follows: using intermediate M4 (100 mg, 0.25 mmol) and tetrahydropyran-4-ol (51 mg, 0.5 mmol) as raw materials, referring to the synthesis method of compound A4, a light yellow compound A7 (83 mg, yield 69%) was obtained.

[0063] 1 H NMR(500MHz, CDCl3)δ9.32(d,J=6.0Hz,1H),7.82(d,J=8.0Hz,1H),7.77–7.61(m,2H),7.50(t, J=7.5Hz,1H),7.44(t,J=7.5Hz,1H),7.15(t,J=6.5Hz,1H),7.06(t,J=6.0Hz,1H),6.44(s,1H) ,5.28(s,1H),4.66–4.58(m,1H),4.57–4.41(m,4H),4.13–4.01(m,2H),3.64(t,J=10.0Hz,2H) ,3.11(d,J=3.0Hz,3H),2.18–2.06(m,2H),1.98–1.81(m,2H).HRMS(ESI-TOF)m / z:[M+H]+calcd for C 27 H 27 N5O4486.2136,found 486.2135.

[0064] The synthesis process of compound A8 is as follows: using intermediate M3a (100 mg, 0.41 mmol) and intermediate M8 (133 mg, 0.49 mmol) as raw materials and referring to the synthesis method of compound A1, a light yellow solid compound A8 (98 mg, yield 50%) was obtained. 1 HNMR (400MHz, CDCl3) δ9.34(d,J=7.2Hz,1H),7.79(d,J=8.0Hz,1H),7.69(d,J=8.8Hz,1H),7.65(d,J =8.4Hz,1H),7.51(t,J=7.2Hz,1H),7.42(t,J=7.2Hz,1H),7.16(t,J=7.2Hz,1H),7.08(t,J=6.0Hz,1H ),7.03(td,J=6.8,0.8Hz,1H),6.49(s,1H),6.10(tt,J=56.0,4.0Hz,1H),4.70–4.60(m,1H),4.56–4. 50(m,4H),4.46(q,J=7.2Hz,2H),4.01–3.86(m,2H),1.48(t,J=7.2Hz,3H).HRMS(ESI-TOF)m / z:[M+H]+ Calculate for C 25 H 23 F2N5O3480.1842, found 480.1845.

[0065] The synthesis process of compound A9 is as follows: using intermediate M3a (100 mg, 0.41 mmol) and intermediate M9 (142 mg, 0.49 mmol) as raw materials and referring to the synthesis method of compound A2, a light yellow solid compound A9 (134 mg, yield 66%) was obtained. 1 HNMR(500MHz, CDCl3)δ9.33(d,J=7.0Hz,1H),7.76(d,J=8.5Hz,1H),7.69(d,J=9.0Hz,1H),7.58–7.50(m,2 H),7.41(td,J=7.0,1.0Hz,1H),7.22(t,J=7.5Hz,1H),7.02(td,J=7.0,1.0Hz,1H),6.57(s,1H),4.74–4.6 2(m,3H),4.62–4.55(m,2H),4.45(q,J=7.0Hz,2H),4.04–3.97(m,1H),3.90–3.80(m,1H),3.22(t,J=5.0Hz ,2H),2.62–2.49(m,2H),2.31(s,3H),2.27–2.21(m,2H),1.47(t,J=7.0Hz,3H).HRMS(ESI-TOF)m / z:[M+H] + Calculate for C 28 H 30 N6O3499.2452, found 499.2447.

[0066] The synthesis process of compound A10 is as follows: intermediate M3a (100 mg, 0.41 mmol) and intermediate M10 (196 mg, 0.49 mmol) are used as raw materials, referring to the synthesis method of compound A2, and then trifluoroacetic acid is used to remove the protecting group to obtain light yellow solid compound A10 (135 mg, 2-step yield 65%). 1H NMR (500MHz, CDCl3) δ9.35(d,J=7.0Hz,1H),7.75(dd,J=11.5,9.0Hz,1H),7.70(d,J=9.0Hz,1H),7.64–7.52(m ,2H),7.42(t,J=8.0Hz,1H),7.25(t,J=8.0Hz,1H),7.03(t,J=7.0Hz,1H),6.58(d,J=18.5Hz,1H),4.79–4.56(m ,5H),4.47(q,J=7.0Hz,2H),4.01–3.61(m,2H),3.23–3.14(m,1H),3.12–2.98(m,2H),2.80(t,J=4.5Hz,1H),1 .49(td,J=7.5,1.5Hz,3H),0.92–0.69(m,2H),0.60–0.48(m,1H),0.33–0.10(m,1H).HRMS(ESI-TOF)m / z:[M+H] + Calculate for C 29 H 30 N6O3511.2452, found 511.2477.

[0067] The synthesis process of compound A11 is as follows: in a 50 mL eggplant-shaped bottle, intermediate M11 (305 mg, 1.0 mmol) and intermediate M3a (245 mg, 1.0 mmol) are dissolved in anhydrous DMF (20 mL), and cesium carbonate (652 mg, 2.0 mmol) is added. The reaction mixture is heated to 110°C for overnight reaction, and the reaction is monitored by TLC. After the reaction is completed, cesium carbonate is filtered off, and the filtrate is concentrated by vacuum distillation. The crude product is purified by silica gel column chromatography to obtain yellow solid compound A11 (272 mg, yield 53%). 1H NMR (400MHz, CDCl3) δ9.35(d,J=6.8Hz,1H),7.99(d,J=8.4Hz,1H),7.75(d,J=8.4Hz,1H),7.69(d,J=8.8H z,1H),7.54(td,J=7.2,1.2Hz,1H),7.42(td,J=7.2,1.2Hz,1H),7.23(t,J=8.0Hz,1H),7.03(td,J=6.8,1. 2Hz,1H),6.84–6.77(m,1H),6.76(s,1H),4.75–4.67(m,1H),4.65–4.59(m,4H),4.46(q,J=7.2Hz,2H),3.6 4–3.55(m,2H),2.71(t,J=6.0Hz,2H),2.58(q,J=7.2Hz,4H),1.48(t,J=7.2Hz,3H),1.03(t,J=7.2Hz,6H).

[0068] The synthesis process of compound A12 is as follows: referring to the synthesis method of compound A1, 2-chloroquinoline is reacted with intermediate M3b to obtain a light yellow solid compound A12 (142 mg, yield 43%), 1 HNMR (400MHz, CDCl3) δ9.22(d,J=6.8Hz,1H),7.90(d,J=8.8Hz,1H),7.77(d,J=8.4Hz,1H),7.62(d,J=7.6Hz,1H),7.55(t,J=7.2Hz,1H),7.23 (t,J=7.6Hz,2H),6.94(t,J=6.8Hz,1H),6.70(d,J=8.8Hz,1H),4.77–4.59(m,5H),4.47(q,J=7.2Hz,2H),2.65(s,3H),1.50(t,J=7.2Hz,3H).

[0069] In this embodiment, the synthetic route of compounds A13 to A20 (see Fig.15 )as follows:

[0070] The synthesis process of compound A13 is as follows: in a 50 mL eggplant-shaped bottle, the intermediate M4 (100 mg, 0.25 mmol) is dissolved in anhydrous N, N-dimethylformamide (10 mL), methylamine hydrochloride (20 mg, 0.30 mmol), 2-(7-azabenzotriazole)-N, N, N', N'-tetramethyluronium hexafluorophosphate (114 mg, 0.30 mmol) and diisopropylethylamine (97 mg, 0.75 mmol) are added in sequence, the reaction mixture is stirred at room temperature overnight, and the reaction is monitored by TLC. After the reaction is completed, the mixture is concentrated by distillation under reduced pressure, and the crude product is purified by silica gel column chromatography to obtain a white solid compound A13 (54 mg, yield 52%). 1 HNMR (500MHz, CD3OD) δ8.85(d,J=7.0Hz,1H),7.89(d,J=8.0Hz,1H),7.69(d,J=8. 0Hz,1H),7.59–7.52(m,2H),7.42(td,J=7.0,1.5Hz,1H),7.25(td,J=7.5,1.0Hz, 1H),7.01(td,J=7.0,1.0Hz,1H),6.77(s,1H),4.62(t,J=8.0Hz,2H),4.58–4.50( m,1H),4.43(t,J=8.0Hz,2H),3.00(s,3H),2.98(s,3H).HRMS(ESI-TOF)m / z:[M+H] + Calculate for C 23 H 22 N6O2415.1877, found415.1875.

[0071] The synthesis process of compound A14 is as follows: using intermediate M4 (100 mg, 0.25 mmol) and 2-methoxyethylamine (23 mg, 0.30 mmol) as raw materials, referring to the synthesis method of compound A13, a light yellow solid compound A14 (54 mg, yield 47%) was obtained. 1HNMR(500MHz,CD3OD)δ8.83(d,J=7.0Hz,1H),7.89(dd,J=8.5,1.0Hz,1H),7.69(d,J=8 .5Hz,1H),7.60–7.52(m,2H),7.43(td,J=7.0,1.0Hz,1H),7.25(td,J=7.0,1.0Hz,1H) ,7.02(td,J=7.0,1.0Hz,1H),6.78(s,1H),4.67–4.61(m,2H),4.62–4.56(m,1H),4.49 –4.42(m,2H),3.65–3.63(m,4H),3.44(s,3H),2.98(s,3H).HRMS(ESI-TOF)m / z:[M+H] + Calculate for C 25 H 26 N6O3459.2139,found 459.2139.

[0072] The synthesis process of compound A15 is as follows: using intermediate M4 (100 mg, 0.25 mmol) and 2,2-difluoroethylamine (24 mg, 0.30 mmol) as raw materials, referring to the synthesis method of compound A13, a light yellow solid compound A15 (66 mg, yield 57%) was obtained. 1 HNMR(500MHz,DMSO-d6)δ8.89(d,J=7.0Hz,1H),8.65(q,J=4.5Hz,1H),8.51(t,J=5.5Hz,1H),7.89(d ,J=8.0Hz,1H),7.65(d,J=9.0Hz,1H),7.62(d,J=8.0Hz,1H),7.55(t,J=7.5Hz,1H),7.43(t,J=7.5Hz, 1H),7.24(t,J=7.5Hz,1H),7.07(t,J=7.0Hz,1H),6.81(s,1H),6.25(tt,J=56.0,3.5Hz,1H),4.63–4. 51(m,3H),4.38(t,J=6.5Hz,2H),3.84–3.73(m,2H),2.84(d,J=4.5Hz,3H).HRMS(ESI-TOF)m / z:[M+H] + Calculate for C 24 H 22 F2N6O2465.1845, found 465.1842.

[0073] The synthesis process of compound A16 is as follows: using intermediate M4 (100 mg, 0.25 mmol) and cyclopropylamine (17 mg, 0.30 mmol) as raw materials, referring to the synthesis method of compound A13, a light yellow solid compound A16 (66 mg, yield 60%) was obtained. 1 HNMR (400MHz, DMSO-d6) δ8.86(d,J=6.8Hz,1H),8.62(d,J=4.4Hz,1H),8.27(d,J=3.6Hz,1H),7.89(d, J=7.6Hz,1H),7.66–7.59(m,2H),7.55(td,J=6.8,1.2Hz,1H),7.39(td,J=6.8,1.2Hz,1H),7.24(td,J =7.2,0.8Hz,1H),7.03(td,J=6.8,0.8Hz,1H),6.81(s,1H),4.55–4.45(m,3H),4.40–4.30(m,2H),2.9 6–2.87(m,1H),2.84(d,J=4.4Hz,3H),0.82–0.74(m,2H),0.71–0.63(m,2H).HRMS(ESI-TOF)m / z:[M+H] + Calculate for C 25 H 24 N6O2441.2034, found 441.2036.

[0074] The synthesis process of compound A17 is as follows: using intermediate M4 (100 mg, 0.25 mmol) and N,N-diethylethylenediamine (35 mg, 0.30 mmol) as raw materials, referring to the synthesis method of compound A13, a light yellow solid compound A17 (82 mg, yield 66%) was obtained. 1 H NMR (500MHz, CD3OD) δ8.92(d,J=7.0Hz,1H),7.90(d,J=8.5Hz,1H),7.70(d,J=8.5Hz,1H),7.58(d ,J=9.0Hz,1H),7.55(d,J=7.5Hz,1H),7.45(t,J=7.5Hz,1H),7.26(t,J=7.5Hz,1H),7.04(t,J=7. 0Hz,1H),6.79(s,1H),4.68–4.59(m,3H),4.47(t,J=5.5Hz,2H),3.61(t,J=7.0Hz,2H),2.98(s,3 H),2.87(t,J=7.0Hz,2H),2.77(q,J=7.0Hz,4H),1.15(t,J=7.0Hz,6H).HRMS(ESI-TOF)m / z:[M+H]+ Calculate for C 28 H 33 N7O2500.2768,found500.2769.

[0075] The synthesis process of compound A18 is as follows: using intermediate M4 (100 mg, 0.25 mmol) and N,N-diethyl-N'-methylethylenediamine (39 mg, 0.30 mmol) as raw materials, referring to the synthesis method of compound A13, a light yellow solid compound A18 (83 mg, yield 65%) was obtained. 1 H NMR (500MHz, CDCl3) δ8.38(d,J=6.5Hz,1H),7.89(d,J=8.5Hz,1H),7.65(d,J=8.0Hz,1H),7.58(d,J=9.0H z,1H),7.50(t,J=7.5Hz,1H),7.25(t,J=8.5Hz,1H),7.19(t,J=7.5Hz,1H),6.95(q,J=4.0Hz,1H),6.86(t, J=6.5Hz,1H),6.56(s,1H),4.54–4.40(m,4H),4.24–4.14(m,1H),3.86–3.39(m,2H),3.09(s,3H),3.06(d ,J=4.5Hz,3H),2.77–2.57(m,2H),2.45(q,J=7.0Hz,4H),0.91(t,J=7.0Hz,6H).HRMS(ESI-TOF)m / z:[M+H] + Calculate for C 29 H 35 N7O2514.2925, found 514.2922.

[0076] The synthesis process of intermediate M13 is as follows: in a 125 mL eggplant-shaped bottle, A11 (272 mg, 0.53 mmol) is dissolved in methanol / water (20 mL / 5 mL), sodium hydroxide (64 mg, 1.59 mmol) is added, the reaction mixture is heated at 65°C and stirred for 3 hours, and the reaction is monitored by TLC. After the reaction is completed, methanol is removed by distillation under reduced pressure to obtain an aqueous solution of the crude product, the pH of the system is adjusted to 4-5 with 5M hydrochloric acid, a solid is precipitated, and the filter cake is washed with water under reduced pressure and dried in air to obtain a light yellow solid M12 (236 mg, yield 92%).

[0077] The synthesis process of compound A19 is as follows: using intermediate M12 (100 mg, 0.21 mmol) and dimethylamine hydrochloride (20 mg, 0.25 mmol) as raw materials, referring to the synthesis method of compound A13, a light yellow solid compound A19 (98 mg, yield 77%) was obtained. 1 HNMR (500MHz, DMSO-d6) δ = 8.61 (t, J = 5.5Hz, 1H), 8.40 (d, J = 7.0Hz, 1H), 7.96 (d, J = 8.5Hz, 1H), 7.66–7 .59(m,2H),7.56(t,J=7.5Hz,1H),7.37(t,J=7.5Hz,1H),7.25(t,J=7.5Hz,1H),7.00(t,J=7.0Hz,1H), 6.78(s,1H),4.55(t,J=8.0Hz,2H),4.38(t,J=8.0Hz,2H),4.25-4.16(m,1H),3.40-3.34(m,2H),3.05( s,6H),2.61(t,J=6.5Hz,2H),2.54(q,J=7.0Hz,4H),0.99(t,J=7.0Hz,6H).HRMS(ESI-TOF)m / z::[M+H] + Calculate for C 29 H 35 N7O2514.2925, found 514.2944.

[0078] The synthesis process of compound A20 is as follows: using intermediate M12 (100 mg, 0.21 mmol) and methoxymethylamine (18 mg, 0.30 mmol) as raw materials, referring to the synthesis method of compound A13, a light yellow solid compound A20 (95 mg, yield 72%) was obtained. 1HNMR (400MHz, CDCl3) δ8.63(d,J=6.8Hz,1H),8.00(d,J=8.4Hz,1H),7.74(d,J=8.4Hz,1H),7.64(d,J=9. 2Hz,1H),7.54(t,J=7.6Hz,1H),7.31(t,J=7.6Hz,1H),7.23(t,J=7.6Hz,1H),6.99–6.92(m,1H),6.90(t ,J=6.8Hz,1H),6.73(s,1H),4.66–4.48(m,4H),4.40–4.30(m,1H),3.62–3.55(m,2H),3.52(s,3H),3.45 (s,3H),2.70(t,J=6.0Hz,2H),2.57(q,J=7.2Hz,4H),1.03(t,J=7.2Hz,6H).HRMS(ESI-TOF)m / z::[M+Na] + Calculate for C 29 H 35 N7O3552.2694, found 552.2693.

[0079] In this example, the synthetic routes of compounds B1 to B6 (see Fig.16 )as follows:

[0080] The synthesis process of intermediates M14a-f is as follows: in a 125 mL eggplant-shaped bottle, intermediate M1 (1.08 mg, 4.0 mmol) is dissolved in acetonitrile (20 mL), and various R3 or R4 substituted 2-aminopyridine P-2-A derivatives (6.0 mmol) and carbon tetrabromide (1.99 g, 6.0 mmol) are added respectively. The reaction mixture is stirred at 80°C overnight, and the reaction is monitored by TLC. After the reaction is completed, the mixture is concentrated by vacuum distillation, and the crude product is purified by silica gel column chromatography to obtain yellow oily intermediate compounds M14a-f respectively.

[0081] Intermediate M14a, yield 67%. 1 H NMR (400MHz, CDCl3) δ9.33(d,J=6.8Hz,1H),7.72(d,J=8.8Hz,1H),7.43(d,J=8.0Hz ,1H),7.03(d,J=6.8Hz,1H),4.50–4.27(m,7H),1.46(s,9H),1.44(d,J=6.8Hz,3H).

[0082] Intermediate M14b, yield 50%. 1H NMR(500MHz, CDCl3)δ9.18(d,J=7.0Hz,1H),7.47(s,1H),6.86(dd,J=7.0,1.5Hz,1H),4.44–4.3 7(m,4H),4.36–4.28(m,2H),4.09–4.03(m,1H),2.46(s,3H),1.45(s,9H),1.43(t,J=7.5Hz,3H).

[0083] Intermediate M14c, yield 19%. 1 H NMR (500MHz, CDCl3) δ9.34 (dd, J=7.5, 5.5Hz, 1H), 7.34 (dd, J=9.0, 2.5Hz, 1H), 6.89 (td, J= 7.5, 2.5Hz, 1H), 4.46–4.37 (m, 4H), 4.36–4.28 (m, 3H), 1.46 (s, 9H), 1.44 (t, J = 7.5Hz, 3H).

[0084] Intermediate M14d, yield 41%. 1 H NMR(500MHz, CDCl3)δ9.15(s,1H),7.61(d,J=9.0Hz,1H),7.28(dd,J=9.0,1.5Hz,1H),4.45–4.3 7(m,4H),4.35–4.28(m,3H),4.09–4.02(m,1H),2.40(s,3H),1.45(s,9H),1.44(t,J=7.5Hz,3H).

[0085] Intermediate M14e, yield 22%. 1 H NMR(400MHz, CDCl3) δ9.32(dd,J=4.8,2.4Hz,1H),7.69(dd,J=10.0,5.2Hz,1H),7.35(td,J =7.6,2.4Hz,1H),4.49–4.37(m,4H),4.37–4.27(m,3H),1.46(s,9H),1.45(t,J=7.2Hz,3H).

[0086] Intermediate M14f, yield 35%. 1H NMR (400MHz, CDCl3) δ8.98 (d, J = 2.4Hz, 1H), 7.60 (d, J = 10.0Hz, 1H), 7.21 (dd, J = 9.6, 2.4Hz, 1H), 4.50–4.36 (m, 4H), 4.35–4.26 (m, 3H), 3.89 (s, 3H), 1.45 (s, 9H), 1.44 (t, J = 7.2Hz, 3H).

[0087] The synthesis process of intermediates M15a-f is as follows: In a 50 mL eggplant-shaped bottle, intermediates 14a-f (1.0 mmol) are dissolved in methanol (10 mL) and water (2 mL), and sodium hydroxide (120 mg, 3.0 mmol) is added. The reaction mixture is stirred at 60°C overnight, and the reaction is monitored by TLC. After the reaction is completed, the aqueous solution of the crude product is obtained by vacuum distillation, and a saturated citric acid solution is added to adjust the pH to 3-4, and the product is extracted with dichloromethane. The organic phase is dried over anhydrous sodium sulfate and concentrated to obtain white solid or light yellow solid intermediate compounds M15a-f (yield 76-88%), which are directly used in the next step without purification.

[0088] The synthesis process of intermediates M16a-f is as follows: in a 50 mL eggplant-shaped bottle, the intermediate compound M15a-f (0.5 mmol) is dissolved in anhydrous N, N-dimethylformamide (10 mL), dimethylamine hydrochloride (DMA·HCl) (1.0 mmol), 2-(7-azabenzotriazole)-N, N, N', N'-tetramethyluronium hexafluorophosphate (228 mg, 0.6 mmol) and diisopropylethylamine (194 mg, 1.5 mmol) are added in sequence, the reaction mixture is stirred at room temperature overnight, and the reaction is monitored by TLC. After the reaction is completed, the crude product is concentrated by vacuum distillation, saturated brine (30 mL) is added thereto, extracted twice with ethyl acetate, dried over anhydrous sodium sulfate, the filtrate is filtered, the crude product is concentrated by vacuum distillation, and purified by silica gel column chromatography to obtain pale yellow or white solid intermediate compounds M16a-f, respectively.

[0089] Intermediate M16a, white solid, yield 92%. 1 H NMR (400MHz, CDCl3) δ9.33(d,J=6.8Hz,1H),7.72(d,J=8.0Hz,1H),7.43(d,J=8.0Hz,1H) ,7.03(d,J=6.8Hz,1H),4.45–4.26(m,4H),3.97–3.84(m,1H),3.09(s,6H),1.46(s,9H).

[0090] Intermediate M16b, white solid, yield 84%. 1H NMR (400MHz, CDCl3) δ8.27 (d, J=6.8Hz, 1H), 7.40 (s, 1H), 6.71 (dd, J=6.8, 1.6Hz, 1H),4.40–4.21(m,4H),3.96–3.86(m,1H),3.09(s,6H),2.42(s,3H),1.45(s,9H).

[0091] Intermediate M16c, light yellow solid, yield 89%. 1 H NMR (400MHz, CDCl3) δ8.42 (dd, J=7.6, 5.6Hz, 1H), 7.29 (dd, J=9.6, 2.4Hz, 1H), 6.76 (t d,J=7.2,2.4Hz,1H),4.39–4.19(m,4H),3.94–3.84(m,1H),3.10(s,6H),1.46(s,9H).

[0092] Intermediate M16d, white solid, yield 98%. 1 HNMR(400MHz, CDCl3)δ8.19(s,1H),7.55(d,J=9.2Hz,1H),7.14(dd,J=9.2,1.6Hz, 1H),4.38–4.20(m,4H),3.98–3.85(m,1H),3.10(s,6H),2.33(s,3H),1.45(s,9H).

[0093] Intermediate M16e, light yellow solid, yield 91%. 1 H NMR (400MHz, CDCl3) δ8.38 (dd, J=4.4, 2.4Hz, 1H), 7.62 (dd, J=10.0, 5.2Hz, 1H), 7.23 ( td,J=8.0,2.4Hz,1H),4.37–4.21(m,4H),3.98–3.85(m,1H),3.10(s,6H),1.46(s,9H).

[0094] Intermediate M16f, white solid, yield 88%. 1 HNMR(400MHz, CDCl3) δ7.98(d,J=2.0Hz,1H),7.53(d,J=10.0Hz,1H),7.08(dd,J=9.6,2 .4Hz,1H),4.34–4.23(m,4H),3.95–3.84(m,1H),3.82(s,3H),3.11(s,6H),1.45(s,9H).

[0095] The synthesis process of intermediates M17a-f is as follows: in a 50 mL eggplant-shaped bottle, compound M16a-f (0.5 mmol) is dissolved in methanol (5 mL), concentrated hydrochloric acid (0.5 mL) is added, the reaction mixture is stirred at room temperature overnight, and the reaction is monitored by TLC. After the reaction is completed, the mixture is concentrated by vacuum distillation to obtain light yellow solid intermediate compounds M17a-f, respectively.

[0096] Intermediate M17a, yield 95%. 1 HNMR (400MHz, CD3OD) δ8.56(d,J=6.8Hz,1H),7.74(d,J=8.0Hz,1H),7.44(d,J=8.0 Hz, 1H), 7.07 (d, J = 6.8 Hz, 1H), 4.75–4.53 (m, 1H), 4.45–4.33 (m, 4H), 3.11 (s, 6H).

[0097] Intermediate M17b, yield 91%. 1 HNMR(400MHz,CD3OD)δ8.57(d,J=6.8Hz,1H),7.85(s,1H),7.46(dd,J=6.8, 1.2Hz,1H),4.74–4.64(m,1H),4.63–4.41(m,4H),3.14(s,6H),2.64(s,3H).

[0098] Intermediate M17c, yield 96%. 1 H NMR (400MHz, CD3OD) δ8.82 (dd, J=7.6, 5.2Hz, 1H), 7.94 (dd, J=8.0, 2.4Hz, 1H), 7.58(td,J=7.6,2.4Hz,1H),4.78–4.66(m,1H),4.65–4.45(m,4H),3.18(s,6H).

[0099] Intermediate M17d, yield 90%. 1 H NMR (500MHz, CD3OD) δ8.15(s,1H),7.57(d,J=9.0Hz,1H),7.36(dd,J=9.0,1.5Hz,1H),4.47–4.34(m,5H),3.09(s,6H),2.37(s,3H).

[0100] Intermediate M17e, yield 96%. 1H NMR (400MHz, CD3OD) δ8.78 (dd, J=7.6, 4.8Hz, 1H), 7.76 (dd, J=7.6, 2.4Hz, 1H), 7.58(td,J=7.6,2.4Hz,1H),4.72–4.63(m,1H),4.67–4.44(m,4H),3.15(s,6H).

[0101] Intermediate M17f, yield 94%. 1 HNMR (400MHz, CD3OD) δ8.19 (d, J=2.0Hz, 1H), 7.96 (d, J=10.0Hz, 1H), 7.84 (dd, J= 10.0,2.4Hz,1H),4.72–4.62(m,1H),4.61–4.41(m,4H),3.97(s,3H),3.16(s,6H).

[0102] In this embodiment, the synthesis process of compounds B1 to B6 is as follows: In a 50 mL eggplant-shaped bottle, intermediates M17a-f (0.25 mmol) and M7 (66 mg, 0.30 mmol) are dissolved in anhydrous DMF (10 mL), and cesium carbonate (244 mg, 0.75 mmol) is added. The reaction mixture is heated to 110 ° C and reacted overnight, and the reaction is monitored by TLC. After the reaction is completed, cesium carbonate is removed by filtration, and the filtrate is distilled under reduced pressure to obtain a crude product, which is purified by silica gel column chromatography to obtain pale yellow or white solid compounds B1 to B6, respectively.

[0103] Compound B1, 75 mg, light yellow solid, yield 77%. 1 H NMR (500MHz, CDCl3) δ8.37 (d, J = 6.0 Hz, 1H), 7.85 (d, J = 7.5 Hz, 1H), 7.61 (d, J = 8. 0Hz,1H),7.57(d,J=9.0Hz,1H),7.48(t,J=7.0Hz,1H),7.32(s,1H),7.26(t,J=8 .0Hz,1H),7.16(t,J=7.0Hz,1H),6.86(t,J=6.0Hz,1H),6.46(s,1H),4.49–4.33 (m,4H),4.10–4.01(m,1H),3.11(s,6H),3.05(s,3H).HRMS(ESI-TOF)m / z::[M+H] + Calculate for C 24 H 24 N6O2429.2034, found 429.2033.

[0104] Compound B2, 38 mg, white solid, yield 69%.1 H NMR (500MHz, CDCl3) δ8.27(d,J=7.0Hz,1H),7.88(d,J=7.5Hz,1H),7.63(d,J=8.5Hz ,1H),7.50(td,J=7.0,1.0Hz,1H),7.33(s,1H),7.18(td,J=7.0,1.0Hz,1H),6.91(q, J=4.5Hz,1H),6.70(dd,J=7.0,1.5Hz,1H),6.53(s,1H),4.50–4.37(m,4H),4.13–4.0 4(m,1H),3.11(s,6H),3.07(d,J=5.0Hz,3H),2.39(s,3H).HRMS(ESI-TOF)m / z:[M+H] + Calculate for C 25 H 26 N6O2443.2190, found 443.2189.

[0105] Compound B3, 50 mg, light yellow solid, yield 72%. 1 H NMR (500MHz, CDCl3) δ8.41(dd,J=7.5,5.5Hz,1H),7.87(d,J=8.0Hz,1H),7.64(d,J=8.0Hz,1H),7.51(d,J=7.0Hz,1H),7.24–7.16(m,2H),6.87(q,J=5. 0Hz,1H),6.75(td,J=7.0,2.5Hz,1H),6.51(s,1H),4.50–4.35(m,4H),4.12 –3.97(m,1H),3.13(s,6H),3.07(d,J=5.0Hz,3H).HRMS(ESI-TOF)m / z:[M+H] + Calculate for C 24 H 23 N6O2F447.1939, found 447.1933.

[0106] Compound B4, 49 mg, white solid, yield 75%. 1H NMR(500MHz, CDCl3)δ8.19(s,1H),7.87(d,J=8.0Hz,1H),7.63(d,J=8.0Hz,1H),7.53–7.45(m,2H),7.17(td,J=7.5,1.0Hz,1H),7.12(dd,J=9.5 ,1.5Hz,1H),6.98(q,J=4.5Hz,1H),6.51(s,1H),4.49–4.35(m,4H),3.12(s,6H),3.06(d,J=5.0Hz,3H),2.32(s,3H).HRMS(ESI-TOF)m / z:[M+H] + Calculate for C 25 H 26 N6O2443.2190, found443.2193.

[0107] Compound B5, 52 mg, light yellow solid, yield 66%. 1 H NMR (400MHz, DMSO) δ8.66 (q, J=4.4Hz, 1H), 8.51 (dd, J=4.4, 2.4Hz, 1H), 7.91 (d, J=8.0Hz, 1H ),7.70(dd,J=10.0,5.2Hz,1H),7.63(d,J=8.0Hz,1H),7.56(td,J=7.2,0.8Hz,1H),7.48(td ,J=8.0,2.4Hz,1H),7.25(t,J=7.2Hz,1H),6.82(s,1H),4.55(t,J=8.0Hz,2H),4.37(t,J=7. 2Hz,2H),4.26–4.15(m,1H),3.05(s,6H),2.85(d,J=4.4Hz,3H).HRMS(ESI-TOF)m / z:[M+Na] + Calculate for C 24 H 23 N6O2F 469.1759,found469.1770.

[0108] Compound B6, 82 mg, white solid, yield 80%. 1H NMR (500MHz, CDCl3) δ7.99(d,J=2.0Hz,1H),7.90(d,J=8.0Hz,1H),7.68(d,J=8.0Hz ,1H),7.52(t,J=7.0Hz,1H),7.48(d,J=9.5Hz,1H),7.22(t,J=7.0Hz,1H),7.06(dd, J=9.5,2.5Hz,1H),6.60(s,1H),6.54(q,J=4.5Hz,1H),4.56–4.42(m,4H),4.16–4.0 6(m,1H),3.82(s,3H),3.15(s,6H),3.08(d,J=5.0Hz,3H).HRMS(ESI-TOF)m / z:[M+H] + Calculate for C 25 H 26 N6O3459.2139, found 459.2142.

[0109] In this example, the synthetic routes of compounds B7 to B22 (see Fig.17 )as follows:

[0110] The synthesis process of intermediate M18 is as follows: In a 125 mL eggplant-shaped bottle, intermediate M17e (2.0 g, 7.62 mmol) and M6 (2.03 g, 9.15 mmol) are dissolved in anhydrous DMF (30 mL), and cesium carbonate (4.96 g, 15.24 mmol) is added. The reaction mixture is heated to 110 ° C and reacted overnight, and the reaction is monitored by TLC. After the reaction is completed, cesium carbonate is removed by filtration, and the filtrate is distilled under reduced pressure to obtain a crude product, which is purified by silica gel column chromatography to obtain a white solid compound M18 (1.64 g, yield 48%). 1 HNMR(500MHz, CDCl3)δ8.47(d,J=8.5Hz,1H),8.41(dd,J=4.0,2.5Hz,1H),7.78(d,J=8.5Hz,1H),7.62–7.55(m,2H),7.31(td,J=7.5,1.0Hz ,1H),7.21(td,J=7.5,2.0Hz,1H),7.17(s,1H),4.62(t,J=8.0Hz,2H),4.57(t,J=7.0Hz,2H),4.24–4.15(m,1H),4.01(s,3H),3.16(s,6H). 13C NMR(126MHz, CDCl3)δ166.89,162.69,157.81,153.40(d,J=237.8Hz),149.01,148.20(d,J=2.1Hz),144.08,136.40,129.88,126.94,1 25.49, 123.38, 119.92, 118.41 (d, J = 25.4Hz), 117.54 (d, J = 8.9Hz), 116.65 (d, J = 2.0Hz), 113.78, 113.45, 110.99, 56.26, 52.58, 28.55.

[0111] The synthesis process of intermediate M19 is as follows: In a 125mL eggplant-shaped bottle, intermediate M18 (1.64g, 3.66mmol) was dissolved in methanol (30mL) and water (6mL), and sodium hydroxide (440mg, 11.0mmol) was added. The reaction mixture was stirred overnight at 60°C, and the reaction was monitored by TLC. After the reaction was completed, the crude product was distilled under reduced pressure to obtain an aqueous solution, and a saturated citric acid solution was added to adjust the pH to 3-4, and the solid was precipitated, filtered, and the filter cake was washed with water. After air drying, a white solid compound M19 (1.46g, yield 92%) was obtained.

[0112] The synthesis process of compounds B7, B8, B10, B11 and B15 is as follows: in a 50 mL eggplant-shaped bottle, compound M19 (100 mg, 0.23 mmol) is dissolved in anhydrous N, N-dimethylformamide (10 mL), and the corresponding amine compound (0.28 mmol), 2-(7-azabenzotriazole)-N, N, N', N'-tetramethyluronium hexafluorophosphate (105 mg, 0.28 mmol) and diisopropylethylamine (89 mg, 0.69 mmol) are added in sequence, and the reaction mixture is stirred at room temperature overnight, and the reaction is monitored by TLC. After the reaction is completed, the crude product is concentrated by vacuum distillation, saturated brine (30 mL) is added thereto, extracted twice with ethyl acetate, dried over anhydrous sodium sulfate, and the filtrate is concentrated by vacuum distillation. The crude product is purified by silica gel column chromatography to obtain pale yellow or white solid compounds B7, B8, B10, B11 and B15, respectively.

[0113] Compound B7, 72 mg, light yellow solid, yield 56%. 1H NMR (400MHz, DMSO) δ8.61(t,J=5.6Hz,1H),8.51(dd,J=4.4,2.4Hz,1H),7.97(d,J=8.0Hz,1H),7.70(dd,J=9.6 ,5.2Hz,1H),7.63(d,J=8.0Hz,1H),7.56(td,J=7.2,1.2Hz,1H),7.48(td,J=8.0,2.4Hz,1H),7.25(td,J=7.2,1 .2Hz,1H),6.78(s,1H),4.55(t,J=8.0Hz,2H),4.36(t,J=7.2Hz,2H),4.26–4.15(m,1H),3.48–3.38(m,2H),3.0 5(s,6H),2.61(t,J=6.8Hz,2H),2.54(dd,J=14.4,7.2Hz,4H),0.99(t,J=7.2Hz,6H).HRMS(ESI-TOF)m / z:[M+H] + Calculate for C 29 H 34 N7O2F532.2831,found532.2860.

[0114] Compound B8, 62 mg, light yellow solid, yield 57%. 1 H NMR (400MHz, DMSO) δ8.65(t,J=5.6Hz,1H),8.50(dd,J=4.8,2.4Hz,1H),7.93(d,J=7.6Hz,1H),7.70( dd,J=10.0,5.2Hz,1H),7.63(d,J=8.0Hz,1H),7.56(td,J=6.8,1.2Hz,1H),7.48(td,J=8.0,2.0Hz,1H ),7.26(t,J=8.0,1.2Hz,1H),6.79(s,1H),4.55(t,J=8.0Hz,2H),4.37(t,J=8.0Hz,2H),4.26–4.14( m,1H),3.52–3.41(m,2H),3.05(s,6H),2.46(t,J=6.8Hz,2H),2.22(s,6H).HRMS(ESI-TOF)m / z:[M+H] + Calculate for C 27 H 30 N7O2F 504.2518, found 504.2544.

[0115] Compound B10, 51 mg, light yellow solid, yield 48%. 1HNMR(500MHz,DMSO-d6)δ9.79(brs,1H),9.28(brs,1H),9.11(t,J=6.0Hz,1H),8.51(d,J=2.0Hz ,1H),8.07(d,J=8.0Hz,1H),7.71(dd,J=10.0,5.0Hz,1H),7.65(d,J=7.5Hz,1H),7.59(t,J=6.5 Hz,1H),7.48(dt,J=8.0,2.0Hz,1H),7.28(t,J=7.0Hz,1H),7.09(s,1H),4.66–4.53(m,2H),4.4 7–4.35(m,2H),4.27–4.21(m,1H),4.20(d,J=6.0Hz,2H),3.05(s,6H).HRMS(ESI-TOF)m / z:[M+H] + calcdfor C 25 H 24 N7O2FS 506.1769,found 506.1770.

[0116] Compound B11, 52 mg, light yellow solid, yield 49%. 1 H NMR (400MHz, CDCl3) δ8.37(ddd,J=4.4,2.4,0.4Hz,1H),7.86–7.79(m,2H),7.59(d,J=8.0Hz,1H),7.55(ddd,J=9.6,5.2,0.4Hz,1H),7.50(td,J=6. 8,1.2Hz,1H),7.24–7.15(m,2H),6.59(s,1H),4.75(d,J=6.0Hz,2H),4.5 2–4.39(m,4H),4.13–4.04(m,1H),3.12(s,6H).HRMS(ESI-TOF)m / z:[M+H] + Calculate for C 26 H 22 N7O3FS2564.1282, found 564.1307.

[0117] Compound B15, light yellow solid, yield 56%. 1H NMR (400MHz, CDCl3) δ8.40(dd,J=4.0,2.4Hz,1H),7.89(d,J=8.4Hz,1H),7.74(d,J=8.4Hz,1H),7.62–7.50(m ,2H),7.30–7.17(m,2H),6.68(s,1H),6.41(d,J=7.2Hz,1H),4.58(t,J=8.0Hz,2H),4.52(t,J=8.0Hz,2H),4.3 1–4.21(m,1H),4.20–4.10(m,1H),3.47(dd,J=14.4,9.6Hz,1H),3.14(s,6H),2.93–2.77(m,4H),2.63(dd,J=1 4.4,4.0Hz,1H),2.13(dd,J=5.6,2.8Hz,1H),1.81–1.66(m,3H),1.62–1.49(m,1H).HRMS(ESI-TOF)m / z:[M+H] + Calculate for C 30 H 32 N7O2F 542.2674, found 542.2680.

[0118] In a 50 mL eggplant-shaped bottle, compound M19 (100 mg, 0.23 mmol) was dissolved in anhydrous N, N-dimethylformamide (10 mL), and the corresponding amine compound (0.28 mmol), 2-(7-azabenzotriazole)-N, N, N', N'-tetramethyluronium hexafluorophosphate (105 mg, 0.28 mmol) and diisopropylethylamine (89 mg, 0.69 mmol) were added in sequence. The reaction mixture was stirred at room temperature overnight, and the reaction was monitored by TLC. After the reaction was completed, the system was concentrated by vacuum distillation to obtain a crude product, saturated brine (30 mL) was added thereto, extracted twice with ethyl acetate, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated by vacuum distillation to obtain a crude product, which was purified by silica gel column chromatography to obtain a light yellow or white solid compound. The obtained compound was dissolved in dichloromethane (20 mL), trifluoroacetic acid (2 mL) was added, the reaction mixture was stirred at room temperature overnight, and the reaction was monitored by TLC. After the reaction was completed, the crude product was distilled under reduced pressure, and a saturated sodium bicarbonate solution was added to adjust the pH to 9-10. Ethyl acetate (10 mL) was added and extracted twice. The organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and the filtrate was filtered and rotary evaporated under reduced pressure. The crude product was purified by silica gel column chromatography to obtain light yellow solids B9, B12-B14, B16 and B18.

[0119] Compound B9, 32 mg, white solid, yield 42%. 1H NMR (500MHz, CD3OD_SPE) δ8.44(dd,J=4.0,2.5Hz,1H),7.91(d,J=8.5Hz,1H),7.71(d,J=8 .5Hz,1H),7.64–7.54(m,2H),7.42(td,J=8.0,2.5Hz,1H),7.27(t,J=7.5Hz,1H),6.88(s, 1H),4.64(t,J=8.0Hz,2H),4.46(t,J=8.0Hz,2H),4.32–4.23(m,1H),3.49–3.37(m,2H),3 .22(dd,J=12.5,6.5Hz,1H),3.14(s,6H),1.20(d,J=6.5Hz,3H).HRMS(ESI-TOF)m / z:[M+H] + Calculate for C 26 H 28 N7O2F490.2361,found490.2356.

[0120] Compound B12, 57 mg, light yellow solid, yield 38%. 1 H NMR (500MHz, CD3OD) δ8.47–8.45(m,1H),7.96(d,J=7.5Hz,1H),7.75(d,J=8.5Hz,1H),7.66(t,J=7.5Hz,1H ),7.63(dd,J=10.0,5.0Hz,1H),7.45(dt,J=8.0,2.5Hz,1H),7.36(t,J=7.5Hz,1H),6.96(s,1H),4.75(t,J= 8.5Hz,2H),4.72–4.66(m,1H),4.58–4.52(m,2H),4.37–4.29(m,1H),3.68(dd,J=12.0,7.0Hz,1H),3.57–3. 50(m,1H),3.49–3.39(m,2H),3.14(s,6H),2.51–2.41(m,1H),2.25–2.16(m,1H).HRMS(ESI-TOF)m / z:[M+H] + Calculate for C 27 H 28 N7O2F 502.2361,found502.2358.

[0121] Compound B13, 52 mg, light yellow solid, yield 38%. 11H NMR (500 MHz, DMSO) δ 8.79 (d, J = 7.5 Hz, 1H), 8.51 (dd, J = 4.5, 2.5 Hz, 1H), 7.84 (d, J = 8.0 Hz, 1H), 7.69 (dd, J = 10.0, 5.5 Hz, 1H), 7.63 (d, J = 8.0 Hz, 1H), 7.57 (td, J = 7.0, 1.0 Hz, 1H), 7.48 (td, J = 7.0, 2.5 Hz, 1H), 7.26 (td, J = 8.0, 1.0 Hz, 1H), 6.78 (s, 1H), 4.56 (t, J = 8.0 Hz, 2H), 4.37 (t, J = 8.0 Hz, 2H), 4.24–4.17 (m, 1H), 4.11–3.99 (m, 1H), 3.25–3.14 (m, 2H), 3.05 (s, 6H), 2.93–2.82 (m, 2H), 2.04–1.93 (m, 2H), 1.66–1.54 (m, 2H). HRMS (ESI-TOF) m / z: [M+H] + calcd for C 28 H 30 N7O2F5 16.2518, found 516.2495。

[0122] Compound B14, 54 mg, pale yellow solid, yield 44%. 1 1H NMR (500 MHz, CD3OD) δ 8.48 (dd, J = 4.0, 2.0 Hz, 1H), 8.01 (d, J = 8.0 Hz, 1H), 7.87–7.80 (m, 2H), 7.66 (dd, J = 9.5, 5.0 Hz, 1H), 7.54 (dt, J = 6.0, 2.5 Hz, 1H), 7.49 (dt, J = 8.0, 2.5 Hz, 1H), 7.15 (s, 1H), 4.97 (t, J = 9.5 Hz, 2H), 4.73 (dd, J = 9.5, 6.0 Hz, 2H), 4.47–4.39 (m, 1H), 4.39–4.31 (m, 1H), 3.65 (dd, J = 12.5, 4.0 Hz, 1H), 3.35 (dt, J = 12.5, 4.0 Hz, 1H), 3.13 (s, 6H), 3.07–2.98 (m, 2H), 2.23–2.15 (m, 1H), 2.13–2.05 (m, 1H), 1.94–1.83 (m, 1H), 1.79–1.69 (m, 1H). HRMS (ESI-TOF) m / z: [M+H] + calcd for C 28 H 30 N7O2F 516.2518, found 516.2514。

[0123] Compound B16, 50 mg, light yellow solid, yield 53%. 1 H NMR (400MHz, CDCl3) δ8.43–8.38(m,1H),7.75(t,J=9.2Hz,1H),7.58(m,3H),7.31–7.18(m,3H ),6.56(d,J=14.0Hz,1H),4.70–4.59(m,1H),4.59–4.49(m,3H),4.22–4.11(m,1H),4.04–3.59 (m,2H),3.19(dd,J=9.6,4.8Hz,1H),3.15(d,J=1.6Hz,6H),3.11–2.98(m,2H),2.84–2.77(m, 1H),0.85–0.67(m,2H),0.54(t,J=6.9Hz,1H),0.35–0.08(m,1H).HRMS(ESI-TOF)m / z::[M+Na] + Calculate for C 29 H 30 N7O2F 550.2337, found 550.2358.

[0124] Compound B18, 45 mg, light yellow solid, yield 47%. 1 H NMR(500MHz,CD3OD)δ8.47–8.41(m,1H),7.74(d,J=8.5Hz,1H),7.69–7.51(m,3H),7.43(td,J =8.0,2.5Hz,1H),7.31(q,J=8.0Hz,1H),6.87–6.73(m,1H),4.79–4.70(t,1H),4.69–4.58(m, 2H),4.46(t,J=7.0Hz,2H),4.33–4.23(m,1H),3.47(t,J=14.0Hz,1H),3.40–3.31(m,1H),3.2 8–3.16(m,2H),3.14(s,6H),3.10–2.77(m,2H),1.38–0.98(m,3H).HRMS(ESI-TOF)m / z:[M+H] + Calculate for C 28 H 30 N7O2F 516.2518, found 516.2505.

[0125] The synthesis process of compound B17 is as follows: in a 50 mL eggplant-shaped bottle, compound B16 (100 mg, 0.19 mmol) is dissolved in methanol (6 mL), and formaldehyde aqueous solution (0.2 mL, 37%) and sodium cyanoborohydride (36 mg, 0.57 mmol) are added. The reaction mixture is stirred at room temperature overnight and the reaction is monitored by TLC. After the reaction is completed, the crude product is concentrated by vacuum distillation, saturated brine (10 mL) is added thereto, extracted twice with ethyl acetate, dried over anhydrous sodium sulfate, and the filtrate is concentrated by vacuum distillation. The crude product is purified by silica gel column chromatography to obtain a light yellow solid compound B17 (82 mg, yield 80%). 1 H NMR(500MHz,CD3OD)δ8.47–8.42(m,1H),7.73(dd,J=11.5,8.5Hz,1H),7.66–7.57(m,3H),7.43(td,J=8.0,2.0H z,1H),7.31(dd,J=14.0,6.5Hz,1H),6.73(d,J=8.5Hz,1H),4.72–4.57(m,2H),4.50–4.41(m,2H),4.34–4.23(m, 1H),4.11–3.83(m,1H),3.82–3.67(m,1H),3.47–3.37(m,1H),3.14(s,6H),3.10–3.01(m,1H),2.80(t,J=5.0Hz ,1H),2.46(d,J=3.5Hz,3H),0.84–0.71(m,2H),0.68–0.57(m,1H),0.41–0.08(m,1H).HRMS(ESI-TOF)m / z:[M+H] + Calculate for C 30 H 32 N7O2F 542.2674,found 542.2669.

[0126] The synthesis process of compound B19 is as follows: using compound B12 (80 mg, 0.16 mmol) as raw material and referring to the synthesis method of compound B17, a light yellow solid compound B19 (48 mg, yield 59%) was obtained. 1H NMR (400MHz, CDCl3) δ8.40(s,1H),7.96(d,J=8.4Hz,1H),7.73(d,J=8.4Hz,1H),7.62–7.50(m,2 H),7.27–7.16(m,2H),7.07(d,J=6.8Hz,1H),6.71(s,1H),4.82–4.70(m,1H),4.57(t,J=8.0Hz, 2H),4.51(t,J=6.8Hz,2H),4.20–4.08(m,1H),3.14(s,6H),3.05–2.97(m,1H),2.87(d,J=10.0H z,1H),2.67–2.63(m,1H),2.50–2.44(m,1H),2.40(s,3H),2.33–2.27(m,1H),1.96–1.77(m,1H).

[0127] The synthesis process of compound B20 is as follows: using compound B18 (100 mg, 0.19 mmol) as raw material and referring to the synthesis method of compound B17, a light yellow solid compound B20 (74 mg, yield 72%) was obtained. 1 H NMR(400MHz, CDCl3)δ8.41(s,1H),7.81–7.72(m,1H),7.64–7.47(m,3H),7.28–7.18(m,2H),6.63–6.51(m,1H),4.72–4.50(m, 5H),4.21–4.10(m,1H),3.32–3.17(m,2H),3.15(s,6H),2.96–2.60(m,2H),2.30(s,3H),2.21–1.92(m,2H),1.20–0.84(m,3H).

[0128] The synthesis process of compound B21 is as follows: using compound B13 (120 mg, 0.23 mmol) as raw material and referring to the synthesis method of compound B17, a light yellow solid compound B21 (92 mg, yield 75%) was obtained. 1H NMR (400MHz, CDCl3) δ8.38 (dd, J=4.0, 2.4Hz, 1H), 7.86 (d, J=8.0Hz, 1H), 7.68 (d,J=8.4Hz,1H),7.60–7.47(m,2H),7.25–7.15(m,2H),6.66(d,J=8.0Hz,1H) ,6.58(s,1H),4.59–4.40(m,4H),4.15–3.99(m,2H),3.12(s,6H),2.91–2.78( m,2H),2.64–2.57(m,2H),2.29(s,3H),2.17–2.11(m,2H),1.75–1.59(m,2H).

[0129] The synthesis process of compound B22 is as follows: using compound B14 (120 mg, 0.23 mmol) as raw material and referring to the synthesis method of compound B17, a light yellow solid compound B22 (96 mg, yield 78%) was obtained. 1 H NMR (400MHz, CDCl3) δ8.40 (dd, J=4.4, 2.4Hz, 1H), 7.95 (d, J=7.6Hz, 1H), 7.74 (d ,J=8.4Hz,1H),7.62–7.52(m,2H),7.28–7.17(m,2H),6.94(s,1H),6.71(s,1H), 4.58(t,J=8.0Hz,2H),4.52(t,J=6.8Hz,2H),4.41–4.33(m,1H),4.20–4.09(m,1 H),3.14(s,6H),2.52(m,4H),2.24(s,3H),1.88–1.71(m,2H),1.72–1.59(m,2H).

[0130] In this example, the synthetic route of compound B23 (see Fig.18 )as follows:

[0131] The synthesis process of intermediate M20 is as follows: in a 125 mL eggplant-shaped bottle, compound 2-(1-tert-butyloxycarbonyl)azetidin-3-yl)-6-fluoroimidazo[1,2-a]pyridine-3-carboxylic acid (3.35 g, 10.0 mmol), N,O-dimethylhydroxylamine (1.46 g, 15.0 mmol), N,N-diisopropylethylamine (3.88 g, 30.0 mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (4.18 g, 11.0 mmol) are dissolved in N,N-dimethylformamide (30 mL). The reaction mixture is stirred at room temperature overnight and the reaction is monitored by TLC. After the reaction is completed, the filtrate is concentrated by distillation under reduced pressure, and the crude product is purified by silica gel column chromatography to obtain a yellow solid intermediate compound M20 (2.27 g, yield 60%). 1 H NMR(400MHz, CDCl3) δ8.61(dd,J=4.6,2.4Hz,1H),7.64(dd,J=9.8,5.0Hz,1H),7.25(ddd,J=10.0,7 .4, 2.0Hz, 1H), 4.28 (d, J = 8.4Hz, 4H), 4.09 (q, J = 7.6Hz, 1H), 3.47 (s, 3H), 3.42 (s, 3H), 1.45 (s, 9H).

[0132] The synthesis process of intermediate M21 is as follows: in a 125 mL eggplant-shaped bottle, at 0°C, M20 (1.82 g, 4.82 mmol) and methyl magnesium bromide (5 mL, 14.44 mmol) are dissolved in tetrahydrofuran (40 mL). The reaction mixture is then stirred at room temperature overnight, and the reaction is monitored by TLC. After the reaction is completed, saturated ammonium chloride solution is added to the reaction mixture at 0°C, the aqueous phase is extracted twice with ethyl acetate, the organic phase is dried over anhydrous sodium sulfate, the filtered filtrate is concentrated by vacuum distillation, and the obtained crude product is purified by silica gel column chromatography to obtain a white solid intermediate compound M21 (1.01 g, yield 63%). 1 HNMR (400MHz, CDCl3) δ9.76 (dd, J=4.8, 2.0Hz, 1H), 7.71 (dd, J=9.6, 5.2Hz, 1H), 7.42(ddd,J=9.8,7.6,2.4Hz,1H),4.56–4.30(m,5H),2.55(s,3H),1.47(s,9H).

[0133] The synthesis process of intermediate M22 is as follows: in a 50 mL eggplant-shaped bottle, compound M21 (200 mg, 0.6 mmol) is dissolved in methanol (10 mL), concentrated hydrochloric acid (1.0 mL) is added, the reaction mixture is stirred at room temperature overnight, and the reaction is monitored by TLC. After the reaction is completed, the mixture is concentrated by vacuum distillation to obtain a white solid intermediate compound M22 (140 mg, yield 100%). 1 HNMR (400MHz, DMSO-d6) δ9.63 (dd, J=5.4, 2.4Hz, 1H), 7.88 (dd, J=9.8, 5.4Hz, 1H), 7.77–7. 69 (m, 1H), 4.52 (p, J = 8.0Hz, 1H), 4.03 (t, J = 7.4Hz, 2H), 3.77 (t, J = 7.8Hz, 2H), 2.52 (s, 3H).

[0134] The synthesis process of compound B23 is as follows: In a 50 mL eggplant-shaped bottle, intermediate M22 (100 mg, 0.43 mmol) and M7 (95 mg, 0.43 mmol) are dissolved in anhydrous DMF (10 mL), and cesium carbonate (420 mg, 1.29 mmol) is added. The reaction mixture is heated to 110 ° C and reacted overnight, and the reaction is monitored by TLC. After the reaction is completed, cesium carbonate is removed by filtration, and the filtrate is distilled under reduced pressure to obtain a crude product, which is purified by silica gel column chromatography to obtain a white solid compound B23 (86 mg, yield 48%). 1 HNMR (400MHz, CDCl3) δ9.78(dd,J=5.2,2.4Hz,1H),7.91(d,J=8.4Hz,1H),7.71(d,J=8.0Hz,1H),7.67(dd,J=10.0,5.2Hz,1H),7.58–7.51( m,1H),7.43–7.38(m,1H),7.23(t,J=7.6Hz,1H),6.64(s,1H),6.35(q,J=4.8Hz,1H),4.68–4.55(m,5H),3.10(d,J=4.8Hz,3H),2.62(s,3H).

[0135] In this example, the synthetic routes of compounds B24 to B26 (see Fig.19 )as follows:

[0136] The synthesis process of intermediate M23 is as follows: in a 100 mL eggplant-shaped bottle, 2-(azetidin-3-yl)-6-fluoroimidazo[1,2-a]pyridine-3-carboxylic acid ethyl ester M14e (400 mg, 1.52 mmol) and 2-chloro-4-methylquinoline are dissolved in toluene (30 mL), followed by the addition of sodium tert-butoxide (438 mg, 4.56 mmol), x-Phos (145 mg, 0.30 mmol) and Pd2(dba)3 (139 mg, 0.15 mmol), and the reaction mixture is heated at 100°C under argon protection to react overnight. After the reaction is completed, the mixture is distilled under reduced pressure to obtain a crude product, which is purified by silica gel column chromatography to obtain a light yellow solid intermediate compound M23 (128 mg, yield 54%). 1 HNMR(400MHz, CDCl3)δ9.33(dd,J=4.4,2.0Hz,1H),7.81–7.72(m,2H),7.65(dd,J=9.6,4.8Hz,1H),7.52(dt,J=6.8,1.2Hz,1H), 7.35–7.29(m,1H),7.26–7.20(m,1H),6.52(s,1H),4.71–4.55(m,5H),4.47(q,J=7.2Hz,2H),2.59(s,3H),1.49(t,J=7.2Hz,3H).

[0137] The synthesis process of intermediate M24 is as follows: In a 50mL eggplant-shaped bottle, intermediate M23 (295mg, 0.73mmol) was dissolved in methanol (10mL) and water (3mL), and sodium hydroxide (88mg, 2.19mmol) was added. The reaction mixture was stirred overnight at 60°C, and the reaction was monitored by TLC. After the reaction was completed, the crude product was distilled under reduced pressure to obtain an aqueous solution, and a 5M hydrochloric acid solution was added to adjust the pH to 3-4, and the solid was precipitated, filtered, and the filter cake was washed with water. After air drying, a light yellow solid intermediate compound M24 (240mg, yield 88%) was obtained.

[0138] The synthesis process of intermediate M25 is as follows: in a 125 mL eggplant-shaped bottle, intermediate M24 (200 mg, 0.53 mmol), tert-butyl 3,6-diazabicyclo[3.1.1]heptane-6-carboxylate (126 mg, 0.64 mmol), N,N-diisopropylethylamine (205 mg, 1.59 mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (242 mg, 0.64 mmol) are dissolved in N,N-dimethylformamide (10 mL). The reaction mixture is stirred at room temperature overnight and the reaction is monitored by TLC. After the reaction is completed, the filtrate is concentrated by distillation under reduced pressure, and the crude product is purified by silica gel column chromatography to obtain a white solid intermediate compound M25 (280 mg, yield 94.6%).1 H NMR (500MHz, CDCl3) δ8.17(s,1H),7.76(dd,J=8.0,4.0Hz,2H),7.62–7.55(m,2H),7.31(t,J=7.5Hz,1H),7.21(ddd,J=10.0,8.0,2. 5Hz,1H),6.46(s,1H),4.71(s,2H),4.61–4.53(m,2H),4.21(s,4H),3.93–3.43(m,3H),2.72–2.66(m,1H),2.56(s,3H),1.49(s,9H).

[0139] The synthesis process of compound B24 is as follows: In a 50 mL eggplant-shaped bottle, intermediate M25 (280 mg, 0.5 mmol) was dissolved in dichloromethane (10 mL), and trifluoroacetic acid (1 mL) was carefully added. The reaction mixture was stirred at room temperature overnight, and the reaction was monitored by TLC. After the reaction was completed, the mixture was distilled under reduced pressure to obtain a crude product, which was purified by silica gel column chromatography to obtain a white solid compound B24 (195 mg, yield 85%) 1 H NMR (400MHz, CDCl3) δ8.27 (dd, J=3.6, 2.4Hz, 1H), 7.79 (dd, J=8.2, 1.2Hz, 1H), 7.75 (d, J=8.0H z,1H),7.59(dd,J=9.8,5.2Hz,1H),7.56–7.51(m,1H),7.25(dd,J=8.2,1.2Hz,1H),7.20(ddd, J=10.0,7.8,2.4Hz,1H),6.50(s,1H),4.60(t,J=8.1Hz,2H),4.56–4.48(m,2H),4.23–4.15(m, 1H), 3.94 (s, 2H), 3.81 (s, 4H), 2.81 (d, J = 9.0Hz, 1H), 2.63–2.51 (m, 3H), 1.63 (d, J = 9.4Hz, 1H).

[0140] The synthesis process of compound B25 is as follows: in a 50 mL eggplant-shaped bottle, compound B24 (143 mg, 0.31 mmol) is dissolved in methanol (10 mL), and formaldehyde aqueous solution (1 mL, 37%) and sodium cyanoborohydride (59 mg, 0.94 mmol) are added. The reaction mixture is stirred at room temperature overnight and the reaction is monitored by TLC. After the reaction is completed, the crude product is concentrated by vacuum distillation, saturated brine (10 mL) is added thereto, extracted twice with ethyl acetate, dried over anhydrous sodium sulfate, and the filtrate is concentrated by vacuum distillation. The crude product is purified by silica gel column chromatography to obtain white solid compound B25 (66 mg, yield 45%). 1H NMR(400MHz, CDCl3)δ8.22(dd,J=4.2,2.2Hz,1H),7.85–7.71(m,2H),7.59(dd, J=9.8,5.2Hz,1H),7.52(ddd,J=8.4,7.0,1.6Hz,1H),7.27–7.12(m,2H),6.50( d,J=1.2Hz,1H),4.60(t,J=8.2Hz,2H),4.56–4.48(m,2H),4.17(ddd,J=14.8,8 .4,6.4Hz,1H),3.92–3.53(m,5H),2.96–2.63(m,3H),2.59(s,3H),2.30(s,3H).

[0141] The synthesis process of compound B26 is as follows: in a 50 mL eggplant-shaped bottle, the intermediate M24 (200 mg, 0.53 mmol) is dissolved in anhydrous N, N-dimethylformamide (10 mL), and N-methylpiperazine (106 mg, 1.06 mmol), 2-(7-azabenzotriazole)-N, N, N', N'-tetramethyluronium hexafluorophosphate (242 mg, 0.64 mmol) and diisopropylethylamine (206 mg, 1.59 mmol) are added in sequence, and the reaction mixture is stirred at room temperature overnight, and the reaction is monitored by TLC. After the reaction is completed, the crude product is concentrated by vacuum distillation, saturated brine (30 mL) is added thereto, extracted twice with ethyl acetate, dried over anhydrous sodium sulfate, and the filtrate is concentrated by vacuum distillation. The crude product is purified by silica gel column chromatography to obtain a light yellow or white solid compound B26 (76 mg, yield 31%). 1 HNMR (400MHz, CDCl3) δ8.36 (dd, J=4.2, 2.4Hz, 1H), 7.79–7.68 (m, 2H), 7.56–7.43 (m, 2H), 7.20–7.11 (m, 2H), 6. 44(s,1H),4.50(dt,J=20.0,7.8Hz,4H),4.18–4.06(m,1H),3.61(s,4H),2.53(s,3H),2.43(s,4H),2.28(s,3H).

[0142] In this example, the synthetic route of compound B27 (see Fig. 20 )as follows:

[0143] The synthesis process of intermediate M26 is as follows: 2,4-dibromoquinoline (1.0 g, 3.48 mmol) was dissolved in anhydrous 1,4-dioxane (20 mL) in a 50 mL eggplant-shaped bottle. The mixture was heated to 90°C, 40% HBr (4 mL) was added dropwise thereto, and the reaction mixture was continuously heated at 90°C overnight. After the reaction was completed, the crude product was concentrated by vacuum distillation, and purified by silica gel column chromatography to obtain an off-white solid intermediate compound M26 (614 mg, yield 79%). 1 H NMR (400MHz, DMSO-d6) δ12.04(s,1H),7.82(dd,J=8.0,1.2Hz,1H),7.61(dt,J=7.2 ,1.2Hz,1H),7.36(dd,J=8.0,0.8Hz,1H),7.30(dt,J=7.2,1.2Hz,1H),7.03(s,1H).

[0144] The synthesis process of intermediate M27 is as follows: in a 100 mL eggplant-shaped bottle, intermediate M26 (224 mg, 1.0 mmol) and 1-methyl-1,2,3,6-tetrahydropyridine-4-boronic acid pinacol ester (223 mg, 1.0 mmol) are dissolved in 1,4-dioxane (20 mL) and H2O (5 mL), followed by the addition of K2CO3 (977 mg, 3.0 mmol) and [1,1'-bis(diphenylphosphino)ferrocene] dichloropalladium Pd(dppf)Cl2 (73 mg, 0.10 mmol), and the reaction mixture is heated at 90°C under argon protection to react overnight. After the reaction is completed, the mixture is distilled under reduced pressure to obtain a crude product, which is purified by silica gel column chromatography to obtain a light yellow solid intermediate compound M27 (192 mg, yield 80%). 1 HNMR(400MHz, CDCl3)δ12.37(s,1H),7.72(d,J=7.6Hz,1H),7.53–7.42(m,2H),7.19(dt,J=8.0,0.8Hz,1 H), 6.55 (s, 1H), 5.84 (s, 1H), 3.25–3.15 (m, 2H), 2.75 (t, J = 5.6Hz, 2H), 2.58–2.51 (m, 2H), 2.48 (s, 3H).

[0145] The synthesis process of intermediate M28 is as follows: in a 100 mL eggplant-shaped bottle, intermediate M27 (575 mg, 2.39 mmol) is dissolved in toluene (20 mL), followed by the addition of phosphorus oxychloride (2 mL), and the reaction mixture is heated at 100°C for 4 hours. After the reaction is completed, the mixture is distilled under reduced pressure to obtain a crude product, which is purified by silica gel column chromatography to obtain a light yellow solid intermediate compound M28 (570 mg, yield 92%). 1HNMR (400MHz, CDCl3) δ7.96(s,1H),7.94(s,1H),7.64(dt,J=7.2,1.2Hz,1H),7.45(dt,J=7.2,1.2Hz,1H),7.14 (s,1H),5.85–5.76(m,1H),3.15(dd,J=6.0,2.8Hz,2H),2.70(t,J=5.6Hz,2H),2.57–2.48(m,2H),2.42(s,3H).

[0146] The synthesis process of compound B27 is as follows: in a 100 mL eggplant-shaped bottle, intermediate M28 (570 mg, 2.20 mmol) and intermediate M17e (578 mg, 2.20 mmol) are dissolved in toluene (30 mL), followed by the addition of sodium tert-butoxide (634 mg, 6.60 mmol), x-Phos (210 mg, 0.44 mmol) and Pd2(dba)3 (201 mg, 0.22 mmol), and the reaction mixture is heated at 100°C under argon protection to react overnight. After the reaction is completed, the mixture is distilled under reduced pressure to obtain a crude product, which is purified by silica gel column chromatography to obtain a light yellow solid compound B27 (156 mg, yield 15%). 1 H NMR(400MHz, CDCl3)δ8.36(dd,J=4.0,2.4Hz,1H),7.74(d,J=0.8Hz,1H),7.72(d,J=0.8Hz,1H),7.56–7.43(m,2H),7.23–7.1 0(m,2H),6.44(s,1H),4.60–4.40(m,4H),4.18–4.06(m,1H),3.76–3.50(m,4H),2.53(s,3H),2.51–2.33(m,4H),2.28(s,3H).

[0147] In this example, the synthetic routes of compounds C1 to C2 (see Fig.21 )as follows:

[0148] The synthesis process of intermediate M29 is as follows: in a 125 mL eggplant-shaped bottle, methyl 2-amino-5-methylbenzoate (1.65 g, 10.0 mmol) is dissolved in acetic acid (10 mL), and an aqueous solution (10 mL) of potassium cyanate (973 mg, 12.0 mmol) is slowly added dropwise, and the reaction mixture is heated at 60°C overnight, and the reaction is monitored by TLC. After the reaction is completed, the reaction is cooled to room temperature, water (40 mL) is added to the reaction mixture, and stirred for 10 minutes, the precipitated solid is filtered, the filter cake is washed with water, and dried in the air to obtain a white solid intermediate compound M29 (1.68 g, yield 81%). 1H NMR (400MHz, DMSO-d6) δ9.57(s,1H),8.26(d,J=8.4Hz,1H),7.69(d,J=1.6Hz,1H),7.33(dd,J=8.8,2.0Hz,1H),6.54(brs,2H),3.86(s,3H),2.26(s,3H).

[0149] The synthesis process of intermediate M30 is as follows: in a 125mL eggplant-shaped bottle, intermediate M29 (1.68g, 8.1mmol) is dissolved in anhydrous ethanol (30mL), sodium hydroxide (645mg, 16.2mmol) is added, the reaction mixture is heated to 80°C and refluxed overnight, and the reaction is monitored by TLC. After the reaction is completed, the temperature is cooled to room temperature, and solids are precipitated in the system, which are filtered, the filter cake is washed with water, and left to dry naturally in the air to obtain a white solid intermediate compound M30 (1.42g, yield 100%). 1 H NMR (400MHz, DMSO-d6) δ9.40 (brs, 1H), 7.47 (s, 1H), 7.09 (dd, J = 8.4, 2.0Hz, 1H), 6.79 (d, J = 8.4Hz, 1H), 2.22 (s, 3H).

[0150] The synthesis process of intermediate M31 is as follows: In a 125mL eggplant-shaped bottle, intermediate M30 (1.42g, 8.1mmol) and phosphorus oxychloride (20mL) are mixed, the reaction mixture is heated to 110°C and refluxed overnight, and the reaction is monitored by TLC. After the reaction is completed, the reaction is cooled to room temperature, and phosphorus oxychloride is removed by vacuum distillation to obtain a crude product. Dichloromethane and triethylamine are slowly added to the cooled crude product until the system pH is 8-9, the mixture is concentrated under reduced pressure, and purified by silica gel column chromatography to obtain a white solid intermediate compound M31 (1.33g, yield 77%). 1 H NMR (400MHz, CDCl3) δ8.03–8.00(m,1H),7.90(d,J=8.8Hz,1H),7.82(dd,J=8.8,2.0Hz,1H),2.61(s,3H).

[0151] The synthesis process of intermediate M32a is as follows: In a 125mL eggplant-shaped bottle, intermediate M31 (891mg, 4.18mmol) is dissolved in anhydrous acetonitrile (40mL) and mixed, and 2,4-dimethoxybenzylamine (699mg, 4.18mmol) and diisopropylethylamine (1.62g, 12.54mmol) are added in sequence. The reaction mixture is heated at 50°C overnight and the reaction is monitored by TLC. After the reaction is completed, the reaction is cooled to room temperature, and solids are precipitated. The precipitated solids are filtered, the filter cake is washed with water, and dried in the air to obtain a white solid intermediate compound M32a (1.32g, yield 92%).1 HNMR(400MHz, CDCl3) δ7.63(d,J=8.4Hz,1H),7.51(dd,J=8.4,1.2Hz,1H),7.35(s,1H),7.33(d,J=8.4Hz,1 H), 6.54–6.45 (m, 2H), 6.25 (t, J = 4.4Hz, 1H), 4.76 (d, J = 5.2Hz, 2H), 3.88 (s, 3H), 3.81 (s, 3H), 2.46 (s, 3H).

[0152] The synthesis process of intermediate M32b is as follows: using intermediate M31 (426 mg, 2.0 mmol) and tert-butyl 3,8-diazabicyclo[3.2.1]octane-3-carboxylate (424 mg, 2.0 mmol) as raw materials, referring to the synthesis method of compound M32a, a white solid intermediate compound M32b (748 mg, yield 97%) was obtained. 1 HNMR(500MHz, CDCl3)δ7.70(d,J=8.5Hz,1H),7.59(s,1H),7.54(dd,J=8.5,1.5Hz,1H),4.57–4 .21(m,4H),3.77–3.42(m,2H),2.49(s,3H),2.00–1.89(m,2H),1.89–1.72(m,2H),1.52(s,9H).

[0153] The synthesis process of intermediate M33a is as follows: In a 50mL eggplant-shaped bottle, intermediate M32a (343mg, 1.0mmol) and M7 (262mg, 1.0mmol) are dissolved in anhydrous DMF (10mL), and cesium carbonate (1.62g, 3.0mmol) is added. The reaction mixture is heated to 110°C for overnight reaction, and the reaction is monitored by TLC. After the reaction is completed, cesium carbonate is removed by filtration, and the filtrate is distilled under reduced pressure to obtain a crude product, which is purified by silica gel column chromatography to obtain a light yellow solid intermediate compound M33a (158mg, yield 28%). 1HNMR (500MHz, CDCl3) δ8.42 (dd, J=4.0, 2.5Hz, 1H), 7.58 (dd, J=10.0, 5.0Hz, 1H), 7.46 (d, J=8. 0Hz,1H),7.34(dd,J=8.5,1.5Hz,1H),7.29(d,J=8.0Hz,1H),7.26(s,1H),7.19(td,J=7.5,2.5 Hz,1H),6.47(d,J=2.5Hz,1H),6.43(dd,J=8.0,2.5Hz,1H),6.06(brs,1H),4.70(d,J=5.5Hz,2 H),4.65–4.52(m,4H),4.13–4.05(m,1H),3.86(s,3H),3.79(s,3H),3.15(s,6H),2.38(s,3H).

[0154] The synthesis process of intermediate M33b is as follows: using intermediates M32b (200 mg, 0.51 mmol) and M7 (135 mg, 0.51 mmol) as raw materials, referring to the synthesis method of compound M33a, a light yellow solid intermediate compound M33b (134 mg, yield 42%) was obtained. 1 H NMR (400MHz, CDCl3) δ8.44–8.39(m,1H),7.59(dd,J=10.0,4.8Hz,1H),7.49(d,J=8.4Hz,1H),7.45(s,1H),7.36(d,J=8.8Hz,1H),7.23–7.16( m,1H),4.60–4.45(m,4H),4.40–4.15(m,4H),4.13–4.01(m,1H),3.62– 3.31(m,2H),3.15(s,6H),2.40(s,3H),1.99–1.85(m,4H),1.51(s,9H).

[0155] The synthesis process of compound C1 is as follows: In a 50 mL eggplant-shaped bottle, the intermediate M33a (158 mg, 0.28 mmol) is dissolved in methanol (10 mL), and concentrated hydrochloric acid (1 mL) is carefully added. The reaction mixture is heated to 65°C for 24 hours, and the reaction is monitored by TLC. After the reaction is completed, the mixture is distilled under reduced pressure to obtain a crude product, which is purified by silica gel column chromatography to obtain a light yellow solid compound C1 (91 mg, yield 78%). 1H NMR (500MHz, DMSO-d6) δ8.49(dd,J=5.0,2.0Hz,1H),7.86(s,1H),7.71(dd,J=10.0,5.5Hz,1H),7.58(brs,2H),7.47(t,J=8.0Hz,1H),7.41(d,J=8.5 Hz,1H),7.27(d,J=8.5Hz,1H),4.43(t,J=8.0Hz,2H),4.27(t,J=8.0Hz,2H ),4.11–4.02(m,1H),3.03(s,6H),2.36(s,3H).HRMS(ESI-TOF)m / z:[M+H] + Calculate for C 22 H 22 N7OF 420.1943,found420.1946.

[0156] The synthesis process of compound C2 is as follows: In a 50 mL eggplant-shaped bottle, the intermediate M33b (134 mg, 0.22 mmol) was dissolved in dichloromethane (10 mL), and trifluoroacetic acid (1 mL) was carefully added. The reaction mixture was stirred at room temperature overnight, and the reaction was monitored by TLC. After the reaction was completed, the mixture was distilled under reduced pressure to obtain a crude product, which was purified by silica gel column chromatography to obtain a light yellow solid compound C2 (88 mg, yield 79%). 1 H NMR(400MHz, CDCl3)δ8.42(s,1H),7.59(dd,J=9.6,5.2Hz,1H),7.48(d,J=8.8Hz,1H), 7.47(s,1H),7.35(d,J=8.8Hz,1H),7.20(t,J=7.6Hz,1H),4.66–4.44(m,4H),4.30–4.1 6(m,2H),4.14–4.02(m,1H),3.64–3.52(m,2H),3.46–3.33(m,2H),3.15(s,6H),2.48( brs,1H),2.40(s,3H),2.22–1.88(m,2H),1.85–1.72(m,2H).HRMS(ESI-TOF)m / z:[M+H] + Calculate for C 28 H 31 N8OF 515.2678, found 515.2682.

[0157] In this example, the synthetic route of compound C3 (see Fig. 22 )as follows:

[0158] The synthesis process of intermediate M34 is as follows: intermediate 2,4-dichloroquinazoline (1.0 g, 5.0 mmol) and tert-butyl 4,7-diazaspiro[2.5]octane-4-carboxylate (1.06 g, 5.0 mmol) are used as raw materials, and referring to the synthesis method of compound M32a, a light yellow solid intermediate compound M34 (1.37 g, yield 73%) is obtained. 1 HNMR(500MHz, CDCl3)δ7.84–7.78(m,2H),7.73(td,J=7.0,1.0Hz,1H),7.42(td,J=7.0,1.0Hz,1H) ,3.94–3.89(m,2H),3.79–3.73(m,4H),1.50(s,9H),1.04(t,J=6.5Hz,2H),0.84(t,J=6.5Hz,2H).

[0159] The synthesis process of intermediate M35 is as follows: using intermediate M34 (187 mg, 0.50 mmol) and M17e (131 mg, 0.50 mmol) as raw materials, referring to the synthesis method of compound M33a, a light yellow solid intermediate compound M35 (71 mg, yield 57%) was obtained. 1 HNMR (400MHz, CDCl3) δ8.41 (dd, J=4.4, 2.4Hz, 1H), 7.65 (d, J=8.0Hz, 1H), 7.61–7.5 5(m,2H),7.54–7.48(m,1H),7.19(td,J=7.6,2.4Hz,1H),7.07(td,J=8.0,1.2Hz,1H) ,4.57(t,J=8.0Hz,2H),4.52(t,J=8.0Hz,2H),4.15–4.04(m,1H),3.78–3.69(m,4H), 3.58(s,2H),3.15(s,6H),1.49(s,9H),1.01(t,J=6.0Hz,2H),0.79(t,J=6.0Hz,2H).

[0160] The synthesis process of compound C3 is as follows: using intermediate M32 (130 mg, 0.22 mmol) as raw material, referring to the synthesis method of compound C2, a light yellow solid compound C3 (93 mg, yield 86%) was obtained. 1HNMR(500MHz, CDCl3)δ8.42(dd,J=4.5,2.5Hz,1H),7.69(d,J=8.0Hz,1H),7.62–7.55(m,2H) ,7.52(t,J=7.5Hz,1H),7.20(td,J=7.5,2.5Hz,1H),7.07(t,J=7.5Hz,1H),4.57(t,J=8.0Hz ,2H),4.52(t,J=8.0Hz,2H),4.15–4.04(m,1H),3.72–3.64(m,2H),3.54(s,2H),3.15(s,6H) ,3.14–3.11(m,2H),0.66(t,J=5.5Hz,2H),0.59(t,J=5.5Hz,2H).HRMS(ESI-TOF)m / z:[M+H] + Calculate for C 27 H 29 N8OF 501.2521, found 501.2518.

[0161] In this example, the synthetic routes of compounds C4 to C8 (see Fig.23 )as follows:

[0162] The synthesis process of intermediate M36 is as follows: using compound M14e (825 mg, 2.27 mmol) as raw material and referring to the synthesis method of compound C1, a yellow solid compound M36 (526 mg, yield 88%) was obtained. 1 HNMR(400MHz,DMSO)δ9.22(dd,J=4.8,2.4Hz,1H),7.88(dd,J=9.6,5.2Hz,1H),7.70(td,J=8.0,2.4Hz,1H),4 .58–4.47(m,1H),4.36(q,J=7.2Hz,2H),4.08(t,J=8.0Hz,2H),3.92(t,J=8.8Hz,2H),1.37(t,J=7.2Hz,3H).

[0163] The synthesis process of compound C4 is as follows: using compound 2-chloro-4-methylquinazoline (357 mg, 2.0 mmol) and M36 (526 mg, 2.0 mmol) as raw materials, referring to the synthesis method of intermediate M33a, a light yellow solid compound C4 (559 mg, yield 69%) was obtained. 1HNMR(400MHz, CDCl3)δ9.33(dd,J=4.8,2.4Hz,1H),7.85(d,J=8.4Hz,1H),7.71–7.56(m,3H),7.32(td,J=7.6,2.4Hz,1H), 7.24–7.15(m,1H),4.75–4.60(m,5H),4.46(q,J=7.2Hz,2H),2.79(s,3H),1.49(t,J=7.2Hz,3H).HRMS(ESI-TOF)m / z:[M+H] + Calculate for C 22 H 20 N5O2F 406.1674, found 406.1675.

[0164] The synthesis process of intermediate M37 is as follows: In a 125mL eggplant-shaped bottle, compound C4 (520mg, 1.28mmol) was dissolved in methanol (20mL) and water (5mL), and sodium hydroxide (154mg, 3.84mmol) was added. The reaction mixture was stirred overnight at 60°C, and the reaction was monitored by TLC. After the reaction was completed, the crude product was distilled under reduced pressure to obtain an aqueous solution, and a saturated citric acid solution was added to adjust the pH to 3-4, and the solid was precipitated, filtered, and the filter cake was washed with water. After air drying, a white solid intermediate compound M37 (460mg, yield 95%) was obtained. 1 HNMR (500MHz, DMSO) δ9.82(dd,J=5.5,2.0Hz,1H),7.99(d,J=8.0Hz,1H),7.68(t,J=7.5Hz,1H),7.55(dd,J=10.0,5.5Hz,1H),7.51(d,J =8.5Hz,1H),7.31(t,J=7.5Hz,1H),7.24(t,J=7.5Hz,1H),5.12–5.00(m,1H),4.45(t,J=7.0Hz,2H),4.34(t,J=7.0Hz,2H),2.75(s,3H).

[0165] The synthesis process of compound C5 is as follows: in a 50 mL eggplant-shaped bottle, the intermediate M37 (100 mg, 0.26 mmol) is dissolved in anhydrous N, N-dimethylformamide (8 mL), and compound methylamine hydrochloride (24 mg, 0.29 mmol), 2-(7-azabenzotriazole)-N, N, N', N'-tetramethyluronium hexafluorophosphate (121 mg, 0.32 mmol) and diisopropylethylamine (103 mg, 0.78 mmol) are added in sequence, and the reaction mixture is stirred at room temperature overnight, and the reaction is monitored by TLC. After the reaction is completed, the crude product is distilled under reduced pressure, and purified by silica gel column chromatography to obtain a light yellow solid C5 (81 mg, yield 76%). 1 H NMR (400MHz, CDCl3) δ8.43(dd,J=4.0,2.4Hz,1H),7.86(d,J=8.4Hz,1H),7.68–7.61(m,2H),7.58(dd,J=10.0,5.2Hz ,1H),7.25–7.14(m,2H),4.72–4.53(m,4H),4.22–4.06(m,1H),3.15(s,6H),2.79(s,3H).HRMS(ESI-TOF)m / z:[M+H] + Calculate for C 22 H 21 N6OF 405.1834,found405.1846.

[0166] The synthesis process of intermediates M38a-c is as follows: in a 50 mL eggplant-shaped bottle, intermediate M37 (100 mg, 0.26 mmol) is dissolved in anhydrous N, N-dimethylformamide (8 mL), various amine compounds (0.29 mmol), 2-(7-azabenzotriazole)-N, N, N', N'-tetramethyluronium hexafluorophosphate (121 mg, 0.32 mmol) and diisopropylethylamine (103 mg, 0.78 mmol) are added in sequence, the reaction mixture is stirred at room temperature overnight, and the reaction is monitored by TLC. After the reaction is completed, the crude product is distilled under reduced pressure, and purified by silica gel column chromatography to obtain yellow solid intermediate compounds M38a-c.

[0167] Intermediate M38a, 126 mg, yield 87%. 1H NMR(400MHz, CDCl3)δ9.32(s,1H),8.02(s,1H),7.88(d,J=8.4Hz,1H),7.70–7.62(m,2H),7.59(dd,J=9.6,5.2Hz,1H),7.31–7.27(m,2H),7.26–7.22 (m,1H),4.83–4.59(m,5H),4.41–4.30(m,1H),3.83–3.75(m,1H),3.58–3. 50(m,2H),2.81(s,3H),2.38–2.27(m,1H),2.10–1.90(m,2H),1.46(s,9H).

[0168] Intermediate M38b, 135 mg, yield 91%. 1 H NMR(500MHz, CDCl3)δ8.95(s,1H),7.92–7.83(m,2H),7.74(t,J=7.5Hz,1H),7.70(dd,J=9.5,5.0Hz,1H),7.35(t,J=7.5Hz,1H),7.29–7.14(m,2 H),4.99–4.83(m,2H),4.79–4.62(m,3H),4.22–4.08(m,3H),3.01–2.88 (m,2H),2.85(s,3H),2.07–1.98(m,2H),1.70–1.57(m,2H),1.46(s,9H).

[0169] Intermediate M38c, 230 mg, 95%. 1 H NMR(500MHz, CDCl3)δ8.43(s,1H),7.87(d,J=8.5Hz,1H),7.68–7.62(m,2H),7.60(dd,J=10.0,5.0Hz,1H),7.25–7.19(m,2H),4.63(t,J=8.5Hz, 2H),4.57(t,J=7.5Hz,2H),4.17–4.08(m,1H),4.07–3.70(m,2H),3.48( m,4H),2.80(s,3H),1.50(s,9H),1.40–1.13(m,2H),1.04–0.48(m,2H).

[0170] Compound C6, 45 mg, yield 82%. 1H NMR (500MHz, CDCl3) δ9.25 (dd, J=4.5, 2.0Hz, 1H), 7.84 (d, J=8.5Hz, 1H), 7.67–7.58 (m, 2H), 7. 56(dd,J=10.0,5.0Hz,1H),7.26–7.17(m,2H),6.61(d,J=7.0Hz,1H),4.77–4.58(m,5H),4.50– 4.39(m,1H),3.26(dd,J=11.5,6.0Hz,1H),3.23–3.15(m,1H),3.08(dd,J=11.5,2.5Hz,1H),3. 04–2.96(m,1H),2.78(s,3H),2.37–2.24(m,1H),1.93–1.80(m,1H).HRMS(ESI-TOF)m / z:[M+H] + Calculate for C 24 H 24 N7OF446.2099, found 446.2098.

[0171] Compound C7, 35 mg, yield 79%. 1 H NMR (400MHz, CDCl3) δ9.24 (dd, J=4.8, 2.4Hz, 1H), 7.85 (d, J=8.0Hz, 1H), 7.68–7.58 (m,2H),7.56(dd,J=10.0,5.2Hz,1H),7.26–7.17(m,2H),6.04(d,J=8.0Hz,1H),4.77 –4.59(m,4H),4.48–4.35(m,1H),4.24–4.10(m,1H),3.30–3.17(m,2H),2.91–2.80( m,2H),2.78(s,3H),2.20–2.08(m,2H),1.72–1.55(m,2H).HRMS(ESI-TOF)m / z:[M+H] + Calculate for C 25 H 26 N7OF 446.2099,found446.2098.

[0172] Compound C8, 48 mg, yield 74%. 1H NMR (400MHz, CDCl3) δ8.42(dd,J=3.6,2.4Hz,1H),7.87(d,J=8.0Hz,1H),7.69–7.61(m,2H),7.58(dd,J=9.6,5.2Hz,1H),7.25–7.16(m ,2H),4.67–4.53(m,4H),4.20–4.08(m,1H),4.04–3.27(m,4H),3.03(s,2H),2.79(s,3H),0.77–0.41(m,4H).HRMS(ESI-TOF)m / z:[M+H] + Calculate for C 26 H 26 N7OF 472.2256,found472.2251.

[0173] In this example, the synthetic routes of compounds C9 to C10 (see Fig.24 )as follows:

[0174] The synthesis process of compound C9 is as follows: 2-chloroquinoxaline (164 mg, 1.0 mmol) and intermediate M3a (245 mg, 1.0 mmol) are dissolved in anhydrous DMF (10 mL) in a 125 mL eggplant-shaped bottle, and cesium carbonate (977 mg, 3.0 mmol) is added. The reaction mixture is heated to 110°C for 6 h, and the reaction is monitored by TLC. After the reaction is completed, cesium carbonate is removed by filtration, and the filtrate is distilled under reduced pressure to obtain a crude product, which is purified by silica gel column chromatography to obtain a light yellow solid compound C9 (186 mg, yield 50%). 1 H NMR (400MHz, CDCl3) δ9.35(d,J=6.8Hz,1H),8.27(s,1H),7.89(dd,J=8.0,1.2Hz,1H),7.75–7.68(m,2H),7.57(dt,J=7.2,1.2Hz,1H),7.43(dt ,J=7.2,1.2Hz,1H),7.38(dt,J=7.2,1.2Hz,1H),7.04(td,J=6.8,1.2Hz,1H),4.82–4.67(m,5H),4.47(q,J=7.2Hz,2H),1.49(t,J=7.2Hz,3H).

[0175] The synthesis process of compound C10 is as follows: using 2-chloroquinoxaline (100 mg, 0.61 mmol) and intermediate M17e (159 mg, 0.61 mmol) as raw materials, referring to the synthesis method of compound C9, a light yellow solid compound C10 (128 mg, yield 54%) was obtained. 1HNMR (400MHz, DMSO-d6) δ8.51(dd,J=4.8,2.4Hz,1H),8.41(s,1H),7.86(d,J=8.8Hz,1H),7.72(dd,J=10.0,5.2Hz,1H),7.66–7.58(m,2H ),7.48(dt,J=6.4,2.0Hz,1H),7.41(dt,J=6.4,2.0Hz,1H),4.65(t,J=8.4Hz,2H),4.44(t,J=8.4Hz,2H),4.30–4.20(m,1H),3.04(s,6H).

[0176] Example Preparation of Compound B16 Hydrochloride or Trifluoroacetate.

[0177] Specifically comprising: dissolving compound B16 in methanol in a 50 mL eggplant-shaped bottle, adding 1.0 mol / L hydrochloric acid aqueous solution under slow stirring, and stirring the reaction mixture at room temperature for 1 hour;

[0178] After the reaction was completed, the system was concentrated by distillation under reduced pressure to obtain a light yellow solid hydrochloride B16·3HCl;

[0179] In a 50 mL eggplant-shaped bottle, compound B16 was dissolved in dry dichloromethane, and trifluoroacetic acid (0.5 mL) was added under slow stirring.

[0180] The reaction mixture was stirred at room temperature for 4 hours;

[0181] After the reaction was completed, the system was concentrated by distillation under reduced pressure to obtain light yellow oily trifluoroacetate B16·CF3COOH.

[0182] Similar organic acid salt products are obtained by reacting the pyrazolopyridine compounds connected by the nitrogen tetracyclic ring with other organic acids, wherein the organic acids include but are not limited to one or more of hydrochloric acid, hydrobromic acid, hydrofluoric acid, phosphoric acid, acetic acid, oxalic acid, sulfuric acid, methanesulfonic acid, salicylic acid, trifluoroacetic acid, trifluoromethanesulfonic acid, naphthalenesulfonic acid, maleic acid, fumaric acid, citric acid, citric acid, acetic acid, tartaric acid, succinic acid, malic acid, and glutamic acid.

[0183] (3) Performance testing of representative compounds

[0184] The following tests were performed on the pyrazolopyridine compounds connected by the nitrogen four-membered rings in each embodiment:

[0185] (3.1) Activity inhibition test of a class of nitrogen-containing four-membered ring-linked pyrazolopyridine compounds on PDE10A (phosphodiesterase type 10A)

[0186] At room temperature, the nitrogen-containing four-membered ring-linked pyrazolopyridine compound to be tested was mixed with 20,000-30,000 cpm containing 1.0 μg / mL recombinant cloned PDE10A protein, 20 mmol / L Tris-HCl (pH = 7.5), 4 mmol / L dithiothreitol, and 10 mmol / LMgCl2. 3 The H-cAMP was mixed and incubated for 15 minutes, and then the reaction was stopped with 0.2 mol / L ZnSO4 and 0.2 mol / L Ba(OH)2, respectively. The unreacted supernatant was measured using a PerkinElmer 2910 counter. 3 H-cAMP, the experiment was repeated at least 3 times. Through concentration testing and nonlinear regression calculation, the IC values ​​of pyrazolopyridine compounds connected by nitrogen tetracyclic rings on the inhibition of recombinant cloned PDE10A protein activity can be obtained. 50 value.

[0187] The results of the inhibitory test of the activity of the pyrazolopyridine compounds connected by the nitrogen tetracyclic rings on the recombinant cloned PDE10A protein in each embodiment are shown in Table 1. Under the same conditions, the inhibitory test results of the positive control (opium vera) on the activity of the recombinant cloned PDE10A protein should be controlled within the IC 50 =50~100nM range to ensure the IC of the tested compound 50 Value data has a reference to a unified standard:

[0188] Table 1 Inhibitory activity of PDE10A protein by pyrazolopyridine compounds linked by nitrogen tetracyclic rings

[0189] Compound <![CDATA[IC 50 (nmol / L)]]> Compound <![CDATA[IC 50 (nmol / L)]]> Compound <![CDATA[IC 50 (nmol / L)]]> A1 1.5±0.2 A20 12.4 B19 6.4±0.3 A2 8.3±1.2 B1 16.8±2.3 B20 11.2±1.2 A3 2.2±0.1 B2 11.2±0.5 B21 5.9±0.1 A4 3.7±0.2 B3 1.8±0.1 B22 4.2±0.1 A5 1.5±0.1 B4 3.7±0.1 B23 0.74±0.20 A6 4.3±0.1 B5 1.0±0.1 B24 43.8±1.3 A7 1.2±0.2 B6 23.1±3.7 B25 96±5.6 A8 2.0±0.1 B7 2.0±0.1 B26 12.1±1.1 A9 11±1.1 B8 1.2±0.1 B27 4.0±0.3 A10 4.7±0.3 B9 3.1±0.4 C1 1.8±0.1 A11 3.5±0.4 B10 3.4±0.2 C2 7.2±0.3 A12 3.5±1.0 B11 0.32±0.02 C3 1.0±0.2 A13 20.4±1.3 B12 4.3±0.7 C4 3.7±1.0 A14 49.3±0.5 B13 7.9±0.6 C5 2.5±0.4 A15 73.5±5.2 B14 3.6±0.1 C6 >100 A16 51.0±3.8 B15 5.5±0.2 C7 8.4±1.5 A17 >100 B16 10.9±1.2 C8 9.3±1.6 A18 >100 B17 5.8±0.3 C9 2.1±0.1 A19 5.9±0.1 B18 6.1±0.6 C10 4.6±0.3

[0190] (3.2) Metabolic stability test of a class of nitrogen-containing four-membered ring-linked pyrazolopyridine compounds

[0191] 1μmol / L of the tested nitrogen-four-membered ring-linked pyrazolopyridine compound was tested with 0.5mg / mL rat liver microsomes (RLM) and 1.0mmol / L NADPH in an incubation system with a medium of 0.1mol / L PBS buffer (pH=7.4) and a temperature of 37℃, and the reaction was stopped by taking 30μL from the incubation system at 0, 5, 15, 30, and 60min, and adding 200μL of pre-cooled methanol (containing 20μL of 1μg / mL tolbutamide); then the content of the remaining nitrogen-four-membered ring-linked pyrazolopyridine compound in the system was measured by Thermo ultra-high performance liquid chromatography tandem mass spectrometry, and the measurement was repeated 3 times. The half-life value of the nitrogen-four-membered ring-linked pyrazolopyridine compound metabolic by rat liver microsomes (RLM) was calculated by concentration logarithmic test and linear regression. The larger the half-life value of the pyrazolopyridine compound connected by the nitrogen four-membered ring after metabolism in rat liver microsomes (RLM), the slower the metabolism of the compound by the CYP metabolizing enzyme in vitro, indicating that the compound may be more stable in vivo.

[0192] The metabolic stability test results of representative nitrogen-containing four-membered ring-connected pyrazolopyridine compounds are shown in Table 2. Under the same conditions, the metabolic stability half-life of the positive control (phenacetin) is T 1 / 2 In the range of 30 to 50 minutes, the half-life T of the tested compound should be guaranteed. 1 / 2 The data has a unified reference standard:

[0193] Table 2 Metabolic stability test results of pyrazolopyridine compounds connected by nitrogen four-membered ring

[0194]

[0195] (3.3) Selective testing of nitrogen-containing four-membered ring-linked pyrazolopyridine compounds for the PDEs superfamily

[0196] Taking nitrogen-containing four-membered ring-linked pyrazolopyridine compounds A11, B11 and B16 as examples, the selectivity of the inhibitory effect of nitrogen-containing four-membered ring-linked pyrazolopyridine compounds on the PDEs family was tested; the specific experimental process refers to (1) the activity inhibition test method of nitrogen-containing four-membered ring-linked pyrazolopyridine compounds on PDE10A (phosphodiesterase type 10A);

[0197] The selectivity test results of the nitrogen-containing four-membered ring-connected pyrazolopyridine compounds A11, B11 and B16 for PDEs family enzymes are shown in Table 3:

[0198] Table 3 Selectivity test results of compounds A11, B11 and B16 for PDEs superfamily

[0199]

[0200]

[0201] Selectivity multiple = IC 50 (PDEs) / IC 50 (PDE10A)

[0202] (3.4) Prediction of blood-brain barrier (BBB) ​​permeability of pyrazolopyridine compounds linked by nitrogen tetracyclic rings

[0203] The nitrogen-four-membered ring-linked pyrazolopyridine compound to be tested was prepared into a stock solution with a concentration of 5 mg / mL using DMSO; 20 μL of the 5 mg / mL stock solution of the nitrogen-four-membered ring-linked pyrazolopyridine compound to be tested was added to a mixture of KH2PO4-K2HPO4 buffer (pH 7.4, 50 mmol / L) and ethanol in a volume ratio of 7:3, and the above was diluted to 100 μg / mL as a secondary stock solution.

[0204] Pig brain tissue extract (PBL, ) and coated on a hydrophobic membrane (0.45μmHydrophobic High Protein Binding Immobilon-P Membrane), then 200μL of 100μg / mL secondary stock solution of the nitrogen-containing four-membered ring-linked pyrazolopyridine compound to be tested was added to the top of the hydrophobic membrane as a dosing pool, and 300μL was added to the other side of the hydrophobic membrane as a receiving pool, keeping the receiving solution in full contact with the hydrophobic membrane. After standing at room temperature for 12 hours, 200μL of 100μg / mL secondary stock solution of the nitrogen-containing four-membered ring-linked pyrazolopyridine compound to be tested was added to 300μL of the mixed solution (pH 7.4, 50mmol / LKH2PO4-K2HPO4 buffer / ethanol = 7:3, V / V), mixed, and used as the theoretical equilibrium concentration solution. The drug concentrations in the receiving cell solution, the theoretical equilibrium concentration solution and the blank well KH2PO4-K2HPO4 buffer were measured using a UV plate reader, and the effective permeability (Pe) of the compound was calculated using Microsoft Office Excel software. Taking the representative nitrogen-containing four-membered ring-linked pyrazolopyridine compounds in Table 4 as an example, the effective permeability Pe was tested. The larger the Pe value, the higher the blood-brain barrier permeability of the compound. The candidate drug for the treatment of schizophrenia, PDE10A selective inhibitor MP-10, was used for comparison. The test results are shown in Table 4:

[0205] Table 4 Blood-brain barrier permeability test of pyrazolopyridine compounds connected by nitrogen tetracyclic rings

[0206]

[0207]

[0208] From Table 4, it can be seen that the effective permeability Pe value of MP-10 is the largest, and when Pe≥2.0×10 -6 cm·s -1 When the compound is connected to a nitrogen tetra-membered ring, it is generally predicted that the compound has good blood-brain barrier permeability. Compared with MP-10, the blood-brain barrier permeability of the pyrazolopyridine compound connected to the nitrogen tetra-membered ring of the present invention is lower, and its potential inhibitory effect on PDE10A in the central nervous system is smaller, indicating that it is very likely to reduce the potential side effects of central nervous system inhibition. Therefore, when the pyrazolopyridine compound connected to the nitrogen tetra-membered ring of the present invention is used to treat and / or prevent related diseases including myocardial hypertrophy, myocardial remodeling, myocardial fibrosis, myocardial injury, etc., it will not have a significant inhibitory effect on PDE10A in the central nervous system.

[0209] (3.5) Confirmatory experiment on the treatment of myocardial hypertrophy with pyrazolopyridine compounds linked by nitrogen tetracyclic rings

[0210] 36 C57 mice were equally divided into 6 groups. Group 1 was the normal control group (control group), Group 2 was the model group, Group 3 was the 2.5 mg / kg compound B16 group (low-dose group), Group 4 was the 5.0 mg / kg compound B16 group (medium-dose group), Group 5 was the 10.0 mg / kg compound B16 group (high-dose group), and Group 6 was the 10 mg / kg positive control group (propranolol group).

[0211] Except for the normal control group (control group) which was given normal saline, the model group and other groups were subcutaneously injected with isoproterenol ISO 5.0 mg / kg / day for 14 consecutive days to induce myocardial hypertrophy and myocardial damage in C57 mice. At the same time, from the first day of modeling, the high-dose group was gavaged with 10.0 mg / kg of compound B16 once a day, the medium-dose group was gavaged with 5.0 mg / kg of compound B16 once a day, the low-dose group was gavaged with 2.5 mg / kg of compound B16 once a day, the positive control group (propranolol group) was gavaged with 10 mg / kg of propranolol once a day, and the normal control group and the model group were gavaged with an equal volume of solvent [1.6% DMSO + 98.4% CMC-Na (0.5%)] once a day for 14 consecutive days.

[0212] 24 hours after the last administration, echocardiography was performed using a Technos MPX ultrasound system (ESAOTE, Italy) in conjunction with an 8.5-MHz image converter. Specifically, after C57 mice were anesthetized with isoflurane, the chest was shaved, and then echocardiography was performed at the level of the cardiac papillary muscle using a two-dimensional ultrasound-guided M-mode curve (probe frequency was 8.5 MHz). The cardiac function indices tested included left ventricular ejection fraction (EF%), left ventricular fraction shortening (FS%), left ventricular end-diastolic volume, and left ventricular end-systolic volume. At the same time, biological samples were taken and the cardiac index and the content of biomarkers in myocardial tissue (atrial natriuretic peptide (ANP) mRNA and β-myosin heavy chain (β-MHC) mRNA expression) were detected. The results are shown in the attached figure. Figures 1 to 5 shown.

[0213] from Figures 1 to 5 It can be seen that compared with the model group, the nitrogen-tetramembered ring-linked pyrazolopyridine compound B16 of the present invention can improve the heart-to-body weight ratio and heart-to-tibia ratio of ISO-induced myocardial hypertrophy and myocardial injury C57 mice, improve the left ventricular end-systolic diameter and left ventricular end-diastolic diameter of the heart, increase the left ventricular ejection fraction and left ventricular shortening fraction of the heart, and reduce the mRNA content of myocardial tissue biomarkers ANP and β-MHC. At the same time, tissue section staining shows that the nitrogen-tetramembered ring-linked pyrazolopyridine compound B16 mentioned in the present invention can significantly improve myocardial hypertrophy and myocardial fibrosis caused by isoproterenol ISO, and has a therapeutic effect of protecting the myocardium.

[0214] (3.6) Confirmatory experiment on the treatment of pulmonary fibrosis with pyrazolopyridine compounds linked by nitrogen tetracyclic rings

[0215] The anti-pulmonary fibrosis efficacy validation experiment of compound B11 was divided into 6 groups: low-dose group (5.0 mg / kg), high-dose group (10.0 mg / kg), dry powder inhalation pulmonary bolus group (B11 net content 0.5 mg / kg), positive control pirfenidone (PFD) oral administration group (300 mg / kg), normal control group and model group. Each group included 13 male SPF-grade C57 mice, aged 6 to 8 weeks, weighing between 18 and 22 g. Except for the normal control group, which used an equal volume of normal saline instead, all other groups were modeled by intratracheal injection of 3 mg / kg bleomycin, and the modeling administration volume was 5 mL / kg. After modeling, the mice were rotated to evenly distribute the drug solution and induce pulmonary fibrosis. Starting from the second day, [2.0% DMSO, 3.0% Solutol ( HS-15) and 95% saline, V / V], after dissolving B11 in the solvent, high-dose B11 H (10.0 mg / kg) and low-dose B11 L (5.0 mg / kg) were intraperitoneally injected, and the dry powder inhalation pulmonary push administration group (B11 net content 0.5 mg / kg) and the positive control CMC-Na (0.5%) mixed pirfenidone gavage administration group (300 mg / kg) were intervened for three consecutive weeks, and the body weight changes were measured every week. Among them, the pulmonary administration of B11 was carried out after the mice were anesthetized and assisted by endotracheal intubation equipment. The normal control and model groups received an equal volume of saline treatment. In order to evaluate the effect of B11 in the treatment of pulmonary fibrosis, the respiratory function of mice was monitored by dynamic whole body recording (WBP method) in the awake state. After the drug treatment, all mice were awake and respiratory function was tested using an animal respiratory system monitoring instrument. Each group was operated in parallel. Respiratory parameter indicators included: airway stenosis index (Penh), end-expiratory pause (EEP), relaxation time (RT) decrease, etc. Representative results are attached. Figure 6 shown.

[0216] Subsequently, the mice were anesthetized and their lung function was monitored (PFT lung function test system): the animals were anesthetized with 0.45% sodium pentobarbital, the trachea was cut open, and a tracheal tube was inserted. The instrument monitored: airway resistance (Resistance Index, RI), dynamic compliance (Dynamic compliance, Cdyn), total lung capacity (Total Lung Capacity, TLC), inspiratory capacity (Inspiratory capacity, IC), forced vital capacity (Forced vital capacity, FVC) and forced expiratory volume 100 in the first second (Forced expiratory volume 100, FEV100). In the PFT method, B11 can improve the abnormalities of airway resistance (RI), dynamic compliance (Cdyn), total lung capacity (TLC), inspiratory capacity (IC), forced vital capacity (FVC) and forced expiratory volume 100 in the first second (FEV100) caused by bleomycin, see the attached Figure 7 .

[0217] The above results show that, whether it is the WBP or PFT method, B11 can improve the decline in lung function in mice with pulmonary fibrosis, and the B11 dry powder inhalation pulmonary administration group (B11 net content 0.5 mg / kg) has the best effect, followed by the B11 intraperitoneal injection high-dose group (10 mg / kg), and both are better than the positive drug PFD group, indicating that the B11 inhalation preparation can be effective at a lower dosage, greatly reducing its potential risk of toxic and side effects.

[0218] The lung tissues from the same part were fixed and stored in 4% paraformaldehyde at room temperature. The lung tissues from the same part (about 10 mg) were stored in RNAlater preservation solution at -80°C. The remaining lung tissues were placed in cryovials and quickly frozen in liquid nitrogen and then stored at -80°C. The lung tissue sections were commissioned to a professional biotechnology company for routine H&E and Masson staining, and semi-quantitative scoring was performed randomly by relevant professionals. Figure 8 As shown in the figure, after B11 administration, H&E staining showed that the degree of alveolitis and pulmonary fibrosis was improved, and Masson staining showed that collagen fiber proliferation was reduced. Semi-quantitative scores showed that the B11 pulmonary administration group (B11 net content 0.5 mg / kg) had the best effect, followed by the B11 intraperitoneal injection high-dose group (10 mg / kg), which were better than the oral administration of pirfenidone PFD group. The above results show that B11 can improve the pulmonary fibrosis lesions of mice with bleomycin-induced pulmonary fibrosis.

[0219] (4) Representative Example Preparation of Dry Powder Inhaler ( Fig. 9 )

[0220] (4.1) In this embodiment, the preparation method of compound B11 nanosuspension is as follows:

[0221] Compound B11 has poor solubility in an aqueous solution containing a lyophilization protectant and cannot achieve the expected drug loading. This problem can be solved by preparing it into a nanosuspension, such as Fig. 9 (Left) As shown. One or more of poloxamer F68, poloxamer F127, sodium dodecyl sulfate, phospholipids, polyvinyl alcohol, sodium cholate, sodium deoxycholate, vitamin E polyethylene glycol succinate (TPGS), Tween 80, Soluplus, etc. are selected as stabilizers for nanosuspensions and prepared into a 0.1-5% aqueous solution, which is recorded as solution A. Compound B11 is prepared into a saturated solution with solvents such as DMSO, acetone, methanol, or ethanol. Use a precision syringe pump to push it into 5-20 times the volume of liquid A, the inner diameter of the PTFE tube is 0.3-1.0mm, the push speed is 0.1-1.0mL / min, the collection container is a 10mL vial, and the stirring speed of the collection bottle liquid is 1000-1500rpm. The nanosuspension is subjected to probe ultrasonic dispersion under ice bath conditions, the ultrasonic power is 20-60W, the total working time is 15-30min, and the ultrasound is turned on for 2s and stopped for 1s. The B11 nanosuspension was purified by centrifugation, ultrafiltration and other methods.

[0222] (4.2) Preparation method of compound B11 dry powder inhaler:

[0223] In this study, ultrasonic spray freeze drying technology was used to prepare dry powder inhalation. One or more of mannitol, sucrose, trehalose, glucose, L-leucine, etc. were selected as freeze-drying protective agents. The final type of freeze-drying protective agent was determined based on the appearance of the dry powder, the sphericity of the dry powder particles, the geometric particle size, the drug content, etc. The B11 nanodispersion containing the freeze-drying protective agent was subjected to ultrasonic spray freeze drying. Fig. 9 (right) As shown. The B11 nanosuspension containing the lyophilized protective agent is connected to the precision syringe pump through a syringe, and a 50mL beaker is used as the receiving container. A stirring bar is added to the beaker to stir the liquid nitrogen and prevent boiling and particle adhesion. The beaker is placed on a heat-insulating sponge board, and a small amount of liquid nitrogen is poured in for precooling for tens of seconds. After the boiling sound of the liquid nitrogen gradually weakens, about 30mL of liquid nitrogen is poured in. Monitor the entire process and ensure that there is enough liquid nitrogen.

[0224] The process parameters of ultrasonic spray freeze drying are: solid content of liquid is 5-30%, liquid supply speed is 0.1-5mL / min, inner diameter of PTFE tube is 0.3-1.0mm, frequency of frequency generator is 40-200kHz, amplitude is 20-80%. The collection container contains liquid nitrogen and the like to provide instantaneous freezing. After the collection is completed, cover the mouth of the beaker with a perforated plastic wrap, slowly evaporate until a small amount of liquid nitrogen remains, put it into the freeze dryer, and dry it at -50℃ and 0.05mbar for 24h. Collect the freeze-dried powder and store it in an environment with relative humidity RH <10%.

[0225] (4.3) Characterization of Compound B11 Nanosuspension ( Fig.10 )

[0226] B11 Nanosuspension particle size determination: Dilute the nanosuspension with secondary water to an appropriate concentration (slightly milky white), and use a nanoparticle size and Zeta potential analyzer (Zetasizer nano 2S) for measurement. Set the material refractive index (materialRI) to 1.469 and the dispersant refractive index (dispersant RI) to 1.330. Repeat the measurement three times for each sample. The results are analyzed using Origin 2021 software. The results are as follows: Fig.10 A.

[0227] Observation of B11 nanoparticle morphology by transmission electron microscopy: Place the copper mesh (carbon film mesh) face up on a metal holder, perform hydrophilic treatment on the membrane surface, and use the floating method to prepare the sample. Float the mesh surface on the droplets of nanosuspension (dilute the nanosuspension to a suitable concentration with TPGS solution). After 1 minute, lift the mesh with tweezers and remove the excess solution with filter paper. Then float the mesh surface on phosphotungstic acid dye solution for 1 minute, and remove the excess dye with filter paper. Use a transmission electron microscope (JEM1400) to find the sample and take pictures at 120kV. The results are as follows: Fig.10 B.

[0228] (4.4) Characterization of Compound B11 Dry Powder Inhaler

[0229] Measurement of drug loading: accurately weigh 5 mg of dry powder, dissolve the dry powder with 100 μL DMSO, and then dilute with 900 μL methanol, measure according to chromatographic conditions, record the peak area, and calculate the concentration of compound B11 using the B11 standard curve to obtain the dry powder drug loading data. The experiment was repeated three times in parallel.

[0230] Scanning electron microscope observation of dry powder morphology: Take a non-conductive adhesive and stick it to a metal plate to prepare a sample. Take a small amount of dry powder sample and blow it evenly onto the adhesive surface with an ear-cleaning bulb. Do not touch the sample surface after preparation to prevent changes in the sample morphology. Use an ion sputtering device (J20 IonSputter Coater) to spray gold for 150s at 5W and 6Pa to avoid obvious static electricity in the porous structure. After preparation, put the sample into an EVO tungsten filament scanning electron microscope (EVO MA10) and take a picture at 10.00kV at an appropriate magnification. The results are as follows: Fig.11 A, its particle size distribution statistics are shown in Fig.11 B. (4.5) In vitro aerosol performance test of compound B11 dry powder inhalation

[0231] According to the pharmacopoeia, the aerodynamic characteristics of fine particles in inhalation preparations can be determined using a next generation impactor (NGI). NGI is a cascade impactor with 7 stages and 1 microporous collector (MOC). The device contains a detachable collection cup located on the same horizontal plane. The dry powder in the adapter passes through the nozzles of each stage with the airflow. The smaller the aerodynamic particle size of the particles, the closer the collection plate is to the airflow outlet. Except for the first stage, the other stages adopt a porous design. The aerodynamic particle size of the particles retained at each stage is mainly determined by the airflow velocity and the aperture of each stage.

[0232] The prepared drug-loaded dry powder was loaded into No. 3 HPMC capsules (stored in a drying cabinet), with an average of 9 mg of dry powder loaded into each capsule. The amount of dry powder loaded was accurately weighed. Silicone oil was sprayed into the metal collection trays (S1 to S7 and MOC), 15 mL of solvent was added to the central collection cup of the pre-separator, and the device was connected. The P1 pipeline and the output pipeline on the flow control valve were connected to the DUSA tube, and the adapter and the generator with capsules (without puncture) were connected to the DUSA tube. The pressure pump was turned on and P1 was adjusted to 4.0 KPa; the DUSA tube was removed, and the entire device was fully connected. Without connecting the adapter, the flow sensor was connected to the artificial throat to test the airflow velocity Q and other parameters (P2, P3, PA). The working parameters were set: the number of working times was 10 times, and the working time was calculated according to formula [1]. The capsule was loaded into the generator, and after confirming the puncture, the generator was loaded into the adapter. The working button was clicked to work. After completing 10 capsules, one measurement was completed. Disassemble the device and use a small volume of solvent to collect the deposited dry powder in the generator, adapter, artificial throat, pre-separator and all collection plates. Use methanol to make the collected liquid constant and dissolve the drug in the silicone oil. Use high performance liquid chromatography to detect the drug content. –APSD Data Analysis Software and Origin2021 were used for computational analysis. The results are as follows Fig.12 .

[0233]

[0234] The above description is only a further embodiment of the present invention, but the protection scope of the present invention is not limited thereto. Any technician familiar with the technical field can make equivalent replacements or changes according to the technical solutions and concepts of the present invention within the scope disclosed by the present invention, which belong to the protection scope of the present invention.

Claims

1. A pyrazolopyridine compound connected by a nitrogen four-membered ring, characterized in that: ； A pharmaceutical composition having the structure shown or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient; Wherein: R1, R2, R3 substituents are independently H, halogen or 18 F.C 1-3 Alkyl, C 1-3 One of alkoxy, difluoromethoxy, difluoroethoxy, trifluoromethyl, trifluoromethoxy, acetyl, and cyano; For quinoxaline (A): , R6, R7 substituted quinoline (B) or R6, R 10 Substituted Quinazolines (C): .

2. The pyrazolopyridine compound connected by a nitrogen four-membered ring according to claim 1, characterized in that: R4 is: , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , One of; R5 is H, C 1-5 Alkyl, isopropyl, -C (1-3) CH2OH, -C (1-3) CH2OCH3, -C(=O)CH3, One of -SO2CH3, -C(=O)OCH3.

3. The pyrazolopyridine compound connected by a nitrogen four-membered ring according to claim 1, characterized in that: Quinoline (B) substituted with R6 or R7 ; R6 may independently be a mono- or di-substituted F atom or 18 One of F atom, Cl atom, hydroxyl group, methyl group, trifluoromethyl group, methoxy group and cyano group; R7 can be independently hydroxy, methyl, trifluoromethyl, C 1-3 Alkoxy, R4, -C(=O)-R4, R8, -CH2-R9, -C(=O)CH2-R9, -NHC(=O)-CH2-R9, wherein R8 is: , , , , , , , , , , , , , , , , , , , , , , One of; R9 is: , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , One of; or R6, R 10 Substituted Quinazolines (C) ; R6 may independently be a mono- or di-substituted F atom or 18 F atom, Cl atom, hydroxyl group, methyl group, trifluoromethyl group, methoxy group, cyano group; R 10 May be independently: methyl, ethyl, isopropyl, C1-3 alkoxy, difluoromethoxy, difluoroethoxy, trifluoromethoxy, cyano, one of R8 and R9; In the class of nitrogen-four-membered ring-linked pyrazolopyridine compounds according to claims 1 to 3, the structural units that do not explicitly indicate the (R) configuration or the (S) configuration, the meanings represented by their structures include different configurations.

4. The use of a class of pyrazolopyridine compounds connected by nitrogen four-membered rings according to claims 1 to 3, characterized in that: The pharmaceutically acceptable salt is a product obtained by reacting a pyrazolopyridine compound connected by an azo four-membered ring having a structure represented by formula (I) with an acid; the acid includes but is not limited to one or more of hydrochloric acid, hydrobromic acid, hydrofluoric acid, phosphoric acid, acetic acid, oxalic acid, sulfuric acid, methanesulfonic acid, salicylic acid, trifluoroacetic acid, trifluoromethanesulfonic acid, naphthalenesulfonic acid, maleic acid, fumaric acid, citric acid, citric acid, acetic acid, tartaric acid, succinic acid, malic acid, and glutamic acid.

5. Use of a class of pyrazolopyridine compounds connected by nitrogen-containing four-membered rings, based on the class of pyrazolopyridine compounds connected by nitrogen-containing four-membered rings according to claims 1 to 4, characterized in that: The nitrogen-containing four-membered ring-linked pyrazolopyridine compound to be tested was prepared into a stock solution with a concentration of 5.0 mg / mL using DMSO; 20 μL of the stock solution of the nitrogen-containing four-membered ring-linked pyrazolopyridine compound to be tested with the above concentration was added to a mixture of KH2PO4−K2HPO4 buffer (pH 7.4, 50 mmol / L) and ethanol in a volume ratio of 7:3, and the compound concentration was diluted to 100 μg / mL as a secondary stock solution; Porcine brain tissue extract (PBL, Avanti®) was dissolved in dodecane and coated on a hydrophobic membrane (0.45μ m Hydrophobic High Protein Binding Immobilon-P Membrane). Then, 200 μL of the secondary stock solution of the above-mentioned test compound at 100 μg / mL was added to the top of the hydrophobic membrane as a dosing reservoir, and 300 μL of the above-mentioned mixed solution for dilution was added to the other side of the hydrophobic membrane as a receiving reservoir, keeping the receiving solution in full contact with the hydrophobic membrane. After standing at room temperature for 12 h, 200 μL of the secondary stock solution of 100 μg / mL nitrogen-four-membered ring-linked pyrazolopyridine compounds to be tested was added to 300 μL of the mixed solution (pH 7.4, 50 mmol / L KH2PO4−K2HPO4 buffer and ethanol volume ratio of 7:3) and mixed well to obtain the theoretical equilibrium concentration solution; The drug concentrations in the receptor cell solution, theoretical equilibrium concentration solution, and blank well KH2PO4−K2HPO4 buffer were determined using a UV plate reader, and the effective permeability (Pe) of the compounds was calculated using Microsoft Office Excel software.

6. The use of a class of nitrogen-containing four-membered ring-linked pyrazolopyridine compounds for myocardial protection according to claims 1 to 4, characterized in that: The compound B16 validation experiment was divided into 6 groups, including: 10.0 mg / kg, 5.0 mg / kg, 2.5 mg / kg, one group each, positive control propranolol 10 mg / kg, normal control group, model group, each group had 6 SPF-grade C57 mice; Except for the normal control group which was given normal saline, the model group and each drug-treated group were subcutaneously injected with isoproterenol ISO 5.0 mg / kg / day for 14 consecutive days to induce myocardial hypertrophy and myocardial injury; From the first day of modeling, the normal control group and the model group were gavaged with an equal volume of solvent [1.6% DMSO + 98.4CMC-Na (0.5%)], and the compound B16 and propranolol were prepared and administered using the solvent. The high-dose group H (10 mg / kg) of B16, the medium-dose group M (5.0 mg / kg) of B16, the low-dose group L (2.5 mg / kg) of B16, and the positive control propranolol group 10 mg / kg were gavaged once a day for 14 consecutive days; 24 h after the last administration, echocardiography was performed using the Technos MPX ultrasound system ESAOTE, Italy, coupled with an 8.5 MHz image converter; After the mice were anesthetized with isoflurane, the chest was shaved, and then echocardiography was performed at the level of the cardiac papillary muscle using a two-dimensional ultrasound-guided M-mode curve with a probe frequency of 8.5 MHz. The cardiac function indicators detected included left ventricular ejection fraction (EF%), left ventricular fraction shortening (FS%), left ventricular end-diastolic volume, and left ventricular end-systolic volume. At the same time, biological samples were taken and the cardiac index and the content of biomarkers in myocardial tissue were detected.

7. Use of a class of nitrogen-four-membered ring-linked pyrazolopyridine compounds according to claims 1 to 4 in treating pulmonary fibrosis, characterized in that: The compound B11 validation experiment was divided into 6 groups: low-dose group (5.0 mg / kg), high-dose group (10.0 mg / kg), dry powder inhalation pulmonary bolus group (B11 net content 0.5 mg / kg), positive control pirfenidone (PFD) oral administration group (300 mg / kg), normal control group and model group. Each group included 13 male SPF-grade C57 mice; Except for the normal control group, which used an equal volume of saline instead, all other groups were injected with 3 mg / kg bleomycin intratracheally for modeling, and the modeling volume was 5 mL / kg. After modeling, the mice were rotated to evenly distribute the drug solution and induce pulmonary fibrosis; Starting from the second day after modeling, the normal control group and the model group were given an equal volume of solvent [2.0% DMSO, 3.0% Solutol (Kolliphor® HS-15) and 95% saline, V / V] by gavage. After B11 was dissolved in the solvent, high-dose B11 H (10.0 mg / kg) and low-dose B11 L (5.0 mg / kg) were intraperitoneally injected, and the dry powder inhalation pulmonary bolus administration group (B11 net content 0.5 mg / kg) and the positive control CMC-Na (0.5%) mixed pirfenidone gavage administration group (300 mg / kg) were given once a day for 21 consecutive days. The pulmonary administration of B11 was performed after the mice were anesthetized with the assistance of endotracheal intubation equipment. 24 hours after the last administration, in order to evaluate the effect of B11 in treating pulmonary fibrosis, all mice were in a conscious state and respiratory function was tested using an animal respiratory system monitoring instrument (WBP method). Respiratory parameter indicators included: airway stenosis index (Penh), end-expiratory pause (EEP), relaxation time (RT) reduction, etc. Subsequently, the pulmonary function of the anesthetized mice was monitored again (PFT pulmonary function testing system): the animals were anesthetized with 0.45% sodium pentobarbital, the trachea was incised, and a endotracheal tube was inserted. The instrument monitoring indicators included: airway resistance (Resistance Index, RI), dynamic compliance (Dynamic compliance, Cdyn), total lung capacity (Total Lung Capacity, TLC), inspiratory capacity (Inspiratory capacity, IC), forced vital capacity (Forced vital capacity, FVC) and forced expiratory volume 100 in the first second (Forced expiratory volume 100, FEV100), etc. At the same time, biological samples were taken and the content of biomarkers in lung tissue was detected.

8. The use of a class of pyrazolopyridine compounds connected by nitrogen four-membered rings according to claims 1 to 4, characterized in that: Preparation method of compound B11 nanosuspension: (1) Compound B11 has poor solubility in an aqueous solution containing a lyoprotectant and cannot achieve the expected drug loading. Preparing it into a nanosuspension can solve this problem. (2) Vitamin E polyethylene glycol succinate (TPGS) is preferably used as a stabilizer for the nanosuspension and dissolved in an aqueous solution containing a lyoprotectant, which is recorded as liquid A. (3) Compound B11 is prepared into a 75 mg / mL concentrated solution using DMSO and pushed into 10 times the volume of liquid A using a precision syringe pump. The outlet is a PTFE tube with an inner diameter of 0.6 mm, the injection speed is 0.1 mL / min, the collection container is a 10 mL vial, and the stirring speed is 1200 rpm; (4) the nanosuspension was dispersed by probe ultrasound in an ice bath, the ultrasound power was 40 W, the total working time was 20 min, and the ultrasound was turned on for 2 s and stopped for 1 s; (5) the nanosuspension was centrifuged at 50,000 × g for 30 min in a high-efficiency centrifuge; (6) the supernatant was discarded, and the precipitate was resuspended to a certain volume with liquid A, and the resuspended liquid was dispersed again by ultrasound in an ice bath to obtain a nanosuspension of compound B11; Preparation method of compound B11 dry powder inhaler: (1) This study used ultrasonic spray freeze drying technology to prepare dry powder inhalation. Mannitol and sucrose were selected as freeze-drying protective agents. The final freeze-drying protective agent dosage was determined based on the appearance of the dry powder, the sphericity of the dry powder particles, the geometric particle size, the drug content, etc., and 10% (w / v) mannitol and 5% (w / v) sucrose aqueous solution were preferred; (2) The B11 nanodispersion containing freeze-drying protective agent was connected to the precision injection pump through a syringe, and a 50 mL beaker was used as a receiving container. A stirring bar was added to the beaker to stir the liquid nitrogen and prevent boiling and particle adhesion; (3) The beaker was placed on a heat-insulating sponge board, and a small amount of liquid nitrogen was poured in for precooling for tens of seconds. After the boiling sound of the liquid nitrogen gradually weakened, about 30 mL of liquid nitrogen was poured in. The whole process was monitored and sufficient liquid nitrogen was ensured; (4) The ultrasonic atomization system was first turned on. The system had fixed parameters of 20W and 100kHz, and then the precision injection pump was turned on. After flow correction, the flow rate was set to 0.5 mL / min, start ultrasonic spray, use liquid nitrogen to freeze the droplets and collect them in a beaker; (5) After the collection is completed, cover the mouth of the beaker with a perforated plastic wrap, slowly evaporate until a small amount of liquid nitrogen remains, put it into a freeze dryer, and dry it at -50°C and 0.05 mbar for 24 hours. If it cannot be placed in the freeze dryer quickly, choose to store it at -80°C for subsequent freeze drying; (6) Collect the freeze-dried product, which is the dry powder inhaler of compound B11, and store it in a drying cabinet for use (RH% <10%).

Images