Application of aza-four-membered ring connected pyrazolopyridine compound in preparation of medicine for treating pulmonary arterial hypertension

Abstract

The invention belongs to the technical field of medicines, and particularly discloses application of an aza-four-membered ring connected pyrazolopyridine compound in preparation of medicines for treating pulmonary arterial hypertension. Compared with the prior art, the pyrazolopyridine PDE10A inhibitor connected with the aza four-membered ring can inhibit the enzymatic activity of PDE10A so as to play a potential role in resisting pulmonary arterial hypertension, so that the invention further provides a new application of the compound, and the compound can be applied to preparation of drugs for treating pulmonary arterial hypertension. The invention also relates to application of the pyrazolopyridine PDE10A inhibitor connected with the aza four-membered ring in preparation of drugs for treating pulmonary arterial hypertension.

Description

Technical Field

[0001] The invention relates to the field of pharmaceutical technology, in particular to the preparation of pyrazolopyridine compounds connected by nitrogen tetracyclic rings, and their application as pharmaceutical molecules for treating pulmonary hypertension. Background Art

[0002] Pulmonary arterial hypertension (PAH) is a malignant pulmonary vascular disease with high morbidity and mortality. The pathological state is that the pulmonary artery pressure is too high and exceeds a certain threshold. The clinical symptoms are dyspnea, fatigue, weakness, reduced exercise tolerance, syncope, and chest pain. The pathogenesis of PAH is very complex. Currently, many drugs are used in clinical practice to regulate and intervene in the levels of intracellular cAMP and cGMP to exert therapeutic effects, such as phosphodiesterase PDE5A inhibitors (sildenafil, tadalafil), soluble guanylate cyclase sGC stimulators (riociguat) to exert therapeutic effects by regulating the intracellular NO / sGC / cGMP signaling pathway; prostacyclin PGI2 analogs (epoprostenol) or prostacyclin PGI2 receptor agonists (selexipag) to exert therapeutic effects by regulating the intracellular PGI2 / cAMP signaling pathway. In theory, inhibiting the enzyme activity of intracellular PDE10A will regulate the levels of cAMP and cGMP to a certain extent, and play a potential anti-PAH role.

[0003] At present, there are very few reports on the use of small molecule selective inhibitors developed for the PDE10A target for the treatment of PAH, but they are almost all concentrated in the treatment of central nervous system diseases. This requires that this type of PDE10A inhibitor must have good blood-brain barrier (BBB) ​​permeability to meet its accumulation in the brain and maintain a certain drug concentration. However, this type of reported PDE10A inhibitor used in the treatment of peripheral tissue diseases (such as PAH) will accumulate in brain tissue due to its good BBB permeability and have a strong inhibitory effect on PDE10A highly expressed in brain tissue, thereby significantly increasing the probability of clinical adverse events. On the other hand, the selective PDE5A inhibitor sildenafil, which is widely used in clinical practice, has rapid metabolism in the body and poor selectivity for PDE6 (~16 times), which can easily cause visual impairment. Another PDE5A selective inhibitor, tadalafil, also has poor selectivity for PDE11 (~25 times), which can easily cause adverse reactions such as muscle pain. Highly selective PDE10A inhibitors have more advantages in the selectivity of the PDEs family, usually more than 100 times. Therefore, developing PDE10A inhibitors with low BBB permeability, high selectivity and high activity for the treatment of pulmonary arterial hypertension has more clinical development value and is more innovative. Since PAH patients need to take medication for life, developing oral PDE10A highly selective inhibitors with good drug-like properties is another major challenge. Summary of the invention

[0004] In order to solve the above technical problems, the present invention provides an application of a pyrazolopyridine compound connected by a nitrogen tetra-membered ring in the preparation of a drug for treating pulmonary hypertension.

[0005] To achieve the above object, the present invention is implemented according to the following technical solutions:

[0006] One of the purposes of the present invention is to provide an application of a pyrazolopyridine compound connected by an aza-four-membered ring in the preparation of a drug for treating pulmonary hypertension, wherein the pyrazolopyridine compound connected by an aza-four-membered ring has a structure shown in formula (I):

[0007]

[0008] 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;

[0009] Quinoxaline R6, R7 substituted quinoline and R6, R 10 Substituted Quinazolines One of;

[0010] R4 is

[0011] One of;

[0012] R5 is H, C 1-5 Alkyl, isopropyl, -C (1-3) CH2OH, -C (1-3) One of CH2OCH3, -C(=O)CH3, -SO2CH3, -C(=O)OCH3;

[0013] R6 is independently a mono- or di-substituted F atom, 18 One of F atom, Cl atom, hydroxyl group, methyl group, trifluoromethyl group, methoxy group and cyano group;

[0014] R7 is independently hydroxy, methyl, trifluoromethyl, C 1-3 One of alkoxy, R4, -C(=O)-R4, R8, -CH2-R9, -C(=O)CH2-R9, -NHC(=O)-CH2-R9;

[0015] R8 is:

[0016] One of;

[0017] R9 is

[0018] One of;

[0019] R 10 are independently methyl, ethyl, isopropyl, C 1-3 One of alkoxy, difluoromethoxy, difluoroethoxy, trifluoromethoxy, cyano, R8 and R9.

[0020] The second object of the present invention is to provide a drug for treating pulmonary hypertension, comprising the pyrazolopyridine compound connected by the nitrogen four-membered ring.

[0021] Compared with the prior art, the nitrogen-four-membered ring-linked pyrazolopyridine PDE10A inhibitor of the present invention can inhibit the enzyme activity of PDE10A, thereby exerting a potential anti-pulmonary hypertension effect. Therefore, the present invention also provides a new use of the compound, namely, the use of the nitrogen-four-membered ring-linked pyrazolopyridine PDE10A inhibitor in the preparation of a drug for the treatment of pulmonary hypertension. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 The measured values ​​of mean pulmonary artery pressure (A) and right ventricular systolic pressure (B) of rats in Example 2 are shown.

[0023] Figure 2 is the ratio of the wall thickness of the rat pulmonary arteriole to the cross-sectional diameter (%) (A) in Example 2

[0024] and right heart hypertrophy index measurements in rats (B).

[0025] Figure 3 The rat pulmonary arteriole pathological tissue sections in Example 2, the upper part is a representative hematoxylin and eosin (H&E) staining, the middle part is a representative Masson staining, and the lower part is a representative right ventricular cross-section wheat germ agglutinin (WGA) staining.

[0026] Figure 4 This is the weight measurement statistics of mice in Example 3.

[0027] Figure 5 The right ventricular systolic pressure of mice measured in Example 3 (A) and the right heart hypertrophy index of mice measured (B).

[0028] Figure 6The ratio of the wall thickness of the mouse pulmonary arteriole to the cross-sectional diameter (%) in Example 3 (A) and the statistics of the individual area of ​​myocardial cells after WGA staining of the right ventricular cross section (B).

[0029] Figure 7 The above are representative WGA staining images of the right ventricle cross section and the lower are representative hematoxylin and eosin (H&E) staining images of the mouse pulmonary arteriole pathological tissue sections in Example 3.

[0030] Figure 8 The synthetic route of compounds A1 to A12.

[0031] Fig. 9 This is the synthetic route of compounds M5~M11.

[0032] Fig.10 The synthetic route of compounds A13 to A20.

[0033] Fig.11 The synthetic route of compounds B1 to B6.

[0034] Fig.12 The synthetic route of compounds B7 to B22.

[0035] Fig.13 The synthetic route of compound B23.

[0036] Fig.14 The synthetic route of compounds B24 to B26.

[0037] Fig.15 The synthetic route of compound B27.

[0038] Fig.16 The synthetic route of compounds C1 to C2.

[0039] Fig.17 This is the synthetic route of compound C3.

[0040] Fig.18 This is the synthetic route of compounds C4~C8.

[0041] Fig.19 This is the synthetic route of compounds C9~C10. DETAILED DESCRIPTION

[0042] In order to make the purpose, technical solution and advantages of the present invention more clearly understood, the present invention is further described in detail below in conjunction with embodiments. The specific embodiments described herein are only used to explain the present invention and are not used to limit the invention.

[0043] Example 1

[0044] This embodiment exemplifies some representative nitrogen-containing four-membered ring-linked pyrazolopyridine PDE10A inhibitors, the structures of which are as follows:

[0045]

[0046]

[0047]

[0048] In this example, the synthetic routes of compounds A1 to A12 (see Figure 8 ):

[0049] 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%). 1 H 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).

[0050] 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 (30mL x 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).

[0051] 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).

[0052] 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%). 1H 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 N4O2 393.1659, found 373.1668.

[0053] In this example, the synthetic routes of intermediates M5 to M11 (see Fig. 9 )as follows:

[0054] 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%).

[0055] 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 H NMR (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).

[0056] 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 H NMR(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).

[0057] 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.

[0058] 1 H 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).

[0059] 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. 1HNMR(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).

[0060] 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).

[0061] 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. 1 H NMR (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).

[0062] 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 N4O4 431.1714,found431.1715.

[0063] 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. 1 HNMR(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, found 430.1888.

[0064] 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).

[0065] 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.

[0066] 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. 1 H 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 N5O3 444.2030, found 444.2032.

[0067] 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. 1HNMR(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 N5O4 458.1823, found 458.1826.

[0068] 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.

[0069] 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 N5O4 486.2136,found 486.2135.

[0070] 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.

[0071] 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. 1HNMR(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.

[0072] 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%). 1 H 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 H30 N6O3 511.2452,found 511.2477.

[0073] 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%). 1 H 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).

[0074] 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 H NMR (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).

[0075] In this embodiment, the synthetic route of compounds A13 to A20 (see Fig.10 )as follows:

[0076] 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 white solid compound A13 (54 mg, yield 52%). 1 H NMR (500MHz, CD3OD) δ8.85(d,J=7.0Hz,1H),7.89(d,J=8.0Hz,1H),7.69(d,J=8.0 Hz,1H),7.59–7.52(m,2H),7.42(td,J=7.0,1.5Hz,1H),7.25(td,J=7.5,1.0Hz,1 H),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 N6O2 415.1877, found 415.1875.

[0077] 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 N6O3 459.2139, found 459.2139.

[0078] 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.

[0079] 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 N6O2 441.2034, found 441.2036.

[0080] 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 N7O2 500.2768,found500.2769.

[0081] 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 N7O2 514.2925, found 514.2922.

[0082] 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%).

[0083] 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.

[0084] 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.

[0085] In this example, the synthetic routes of compounds B1 to B6 (see Fig.11 )as follows:

[0086] 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.

[0087] 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).

[0088] 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).

[0089] 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).

[0090] 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).

[0091] 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).

[0092] 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).

[0093] 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.

[0094] 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.

[0095] 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).

[0096] 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).

[0097] 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).

[0098] Intermediate M16d, white solid, yield 98%. 1 H NMR(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).

[0099] 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).

[0100] Intermediate M16f, white solid, yield 88%. 1 H NMR (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).

[0101] 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.

[0102] Intermediate M17a, yield 95%. 1 H NMR (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).

[0103] Intermediate M17b, yield 91%. 1 H NMR (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).

[0104] 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).

[0105] Intermediate M17d, yield 90%. 1 H NMR (500MHz, CD3OD) δ8.15 (s, 1H), 7.57 (d, J = 9.0 Hz, 1H), 7.36 (dd, J = 9.0, 1.5 Hz, 1H), 4.47–4.34 (m, 5H), 3.09 (s, 6H), 2.37 (s, 3H).

[0106] 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).

[0107] Intermediate M17f, yield 94%. 1 H NMR (400MHz, CD3OD) δ8.19(d,J=2.0Hz,1H),7.96(d,J=10.0Hz,1H),7.84(dd,J=1 0.0,2.4Hz,1H),4.72–4.62(m,1H),4.61–4.41(m,4H),3.97(s,3H),3.16(s,6H).

[0108] 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.

[0109] 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 N6O2 429.2034, found 429.2033.

[0110] 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 N6O2 443.2190, found 443.2189.

[0111] 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 N6O2F 447.1939, found 447.1933.

[0112] 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,found 443.2193.

[0113] 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,found 469.1770.

[0114] 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 N6O3 459.2139, found 459.2142.

[0115] In this example, the synthetic routes of compounds B7 to B22 (see Fig.12 )as follows:

[0116] 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 H NMR (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). 13CNMR(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.

[0117] 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.

[0118] 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.

[0119] 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, found 532.2860.

[0120] 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.

[0121] Compound B10, 51 mg, light yellow solid, yield 48%. 1H NMR(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=6.5H z,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),4.47 –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.

[0122] 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 N7O3FS2 564.1282, found 564.1307.

[0123] 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.

[0124] 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.

[0125] 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.

[0126] 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.

[0127] 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 N7O2F516.2518, found 516.2495。

[0128] 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。

[0129] 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.

[0130] 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.

[0131] 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.

[0132] 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).

[0133] 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).

[0134] 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).

[0135] 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).

[0136] In this example, the synthetic route of compound B23 (see Fig.13 )as follows:

[0137] 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).

[0138] 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 H NMR (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).

[0139] 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 H NMR(400MHz, DMSO-d6)δ9.63(dd,J=5.4,2.4Hz,1H),7.88(dd,J=9.8,5.4Hz,1H),7.77–7.6 9(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).

[0140] 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 H NMR (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).

[0141] In this example, the synthetic routes of compounds B24 to B26 (see Fig.14 )as follows:

[0142] 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).

[0143] 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.

[0144] 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).

[0145] 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).

[0146] 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).

[0147] 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).

[0148] In this example, the synthetic route of compound B27 (see Fig.15 )as follows:

[0149] 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).

[0150] 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 H NMR (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).

[0151] 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%). 1H NMR (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).

[0152] 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).

[0153] In this example, the synthetic routes of compounds C1 to C2 (see Fig.16 )as follows:

[0154] 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).

[0155] 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).

[0156] 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).

[0157] 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 H NMR (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,1H ),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).

[0158] 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 H NMR (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).

[0159] 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%). 1H NMR (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).

[0160] 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).

[0161] 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.

[0162] 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.

[0163] In this example, the synthetic route of compound C3 (see Fig.17 )as follows:

[0164] 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 H NMR (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).

[0165] 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 H NMR (400MHz, CDCl3) δ8.41 (dd, J=4.4, 2.4Hz, 1H), 7.65 (d, J=8.0Hz, 1H), 7.61–7.55 (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).

[0166] 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. 1H NMR(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.

[0167] In this example, the synthetic routes of compounds C4 to C8 (see Fig.18 )as follows:

[0168] 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 H NMR (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).

[0169] 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. 1H NMR(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] + calcdfor C 22 H 20 N5O2F 406.1674, found 406.1675.

[0170] 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 H NMR (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).

[0171] 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 HNMR(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=10.0,5.2H z,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.

[0172] 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.

[0173] 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).

[0174] 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).

[0175] 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).

[0176] 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.

[0177] 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, found 446.2098.

[0178] 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, found 472.2251.

[0179] In this example, the synthetic routes of compounds C9 to C10 (see Fig.19 )as follows:

[0180] 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).

[0181] 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).

[0182] The following tests were performed on the pyrazolopyridine compounds connected by the nitrogen four-membered rings in each embodiment:

[0183] Activity inhibition test of pyrazolopyridine compounds linked by nitrogen tetracyclic rings on PDE10A (phosphodiesterase type 10A)

[0184] 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 mM Tris-HCl (pH = 7.5), 4.0 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 (phosphodiesterase type 10A) protein activity can be obtained. 50 value.

[0185] 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 =50~100nmol / L to ensure the IC 50 Value data has a reference to a unified standard.

[0186] Table 1 Inhibitory activity of PDE10A protein by pyrazolopyridine compounds linked by nitrogen tetracyclic rings

[0187]

[0188]

[0189] Example 2: Confirmatory experiment on the treatment of pulmonary hypertension with PDE10A inhibitor in monocrotaline-induced PAH animal model

[0190] Experimental content: Taking the PDE10A inhibitor sample A11 as an example, the experiment was divided into 6 groups (1.25, 2.5, 5.0 mg / kg A11 group, positive control Tadalafil 5.0 mg / kg group, model group, normal control group), with 12 rats in each group. Except for the normal control group given normal saline, the rats in the other groups were intraperitoneally injected with 2% monocrotaline 60 mg / kg once to establish the pulmonary hypertension model. The drug administration started on the second day: (1) The normal control group and the model group were given a blank solvent [1.6% DMSO + 98.4% CMC-Na (0.5%)] by gavage every day; (2) The A11 test group was orally administered 1.25 mg / kg (low-dose group), 2.5 mg / kg (medium-dose group), and 5.0 mg / kg (high-dose group) every day; (3) All animals were given the drug for three weeks, with an oral administration volume of 0.5 mL / 100 g and an intraperitoneal injection volume of 0.1 mL / 100 g. The body weight of the rats was measured every three days during the experiment, and the general state of the rats (respiration, activity, etc.) was observed. One hour after the last administration, the animals were anesthetized with 3% sodium pentobarbital intraperitoneally, and the pulmonary artery pressure of the rats was measured by right cardiac catheterization.

[0191] Determination of rat pulmonary artery pressure (mPAP): After intraperitoneal anesthesia, the rat was fixed in supine position, a longitudinal incision was made from the right clavicle of the neck, the surrounding tissues were bluntly separated, and the right external jugular vein was carefully peeled out with mosquito hemostatic forceps. The vein of about 1 cm was freed, and two lines were led under it. The distal line was used to ligate the vein, and a loose knot was tied on the proximal line for standby use. The proximal and distal lines were fixed on the skin of the rat's chest and neck with hemostatic forceps to fully expose and fill the vein. Adjust and calibrate the physiological recorder, while keeping the sensor parallel to the position of the rat's heart. Use ophthalmic scissors to cut a V-shaped incision at the proximal end, insert the curved end of the right cardiac catheter into the vascular incision, use the proximal line slipknot to cover the catheter inserted into the blood vessel, fix it slightly, and slowly push the catheter. Insert 1 to 2 cm to reach the superior vena cava, 2 to 3 cm to reach the right atrium, gently rotate and push forward, insert about 3.5 to 4 cm to enter the right ventricle, and measure the right ventricular systolic pressure (RVSP). At this time, pause slightly, and then gently push the catheter to enter the pulmonary artery. During the intubation process, the position of the catheter tip should be judged according to the migration change of the pressure curve waveform displayed on the monitor, and the average pulmonary artery pressure should be calculated by the pulmonary artery systolic pressure and pulmonary artery diastolic pressure values.

[0192] Right heart hypertrophy index (RVHI): After the determination of pulmonary hemodynamic indicators, cut the skin of the neck and chest to expose the trachea, quickly dissect the chest cavity to expose the heart and lungs, clamp the trachea with hemostatic forceps, and remove the heart, thymus, and lungs after cutting. Remove the heart, blood vessels, and atrium, free the right ventricle (RV), and leave the left ventricle and ventricular septum (LV+S). Wash the blood with saline and absorb the moisture with filter paper, weigh the right ventricle (RV) and the left ventricle plus the ventricular septum (LV+S), and use the RV / (LV+S) ratio as the right heart hypertrophy index.

[0193] Compared with the model group, the nitrogen tetracyclic ring-linked pyrazolopyridine PDE10A inhibitor A11 of the present invention can significantly reduce the mean pulmonary artery pressure and right heart systolic pressure in rats with pulmonary hypertension induced by monocrotaline (see Figure 1 ), while improving the intimal hyperplasia (thickening) of the pulmonary arteriolar wall caused by pulmonary hypertension, and significantly improving right ventricular hypertrophy caused by pulmonary hypertension (see Figure 2 ). Staining of lung tissue sections included: hematoxylin-eosin H&E, Masson and α-smooth actin α-SMA (see Figure 3 ), further illustrating that the nitrogen-four-membered ring-linked pyrazolopyridine PDE10A inhibitor of the present invention can improve the pathological remodeling of pulmonary arteriolar vessels and exert the pharmacological effect of treating pulmonary hypertension.

[0194] Example 3: Confirmatory Experiment on the Treatment of Pulmonary Hypertension with PDE10A Inhibitors in PAH Animal Model under Hypoxic Environment

[0195] 1. Animal experiments and administration routes

[0196] Taking the PDE10A inhibitor sample A11 as an example, C57 mice (20g, 56 mice) were adaptively raised for three days before the experiment. Afterwards, they were randomly divided into normoxic control group, SU5416 (Sugen, synthetic, purity>99%) + hypoxia (model group), SU5416 + hypoxia medication group (tadalafil group, low, medium and high dose groups of A11) under normoxic conditions (model group). On the first day, the mice in the model group and each medication group were intraperitoneally injected with 20mg / kg of SU5416 and then raised in a hypoxic environment (21 days). During this period, SU5416 (20mg / kg) was injected once on the 8th and 16th days, and the normobaric oxygen control group was raised for 21 days. All mice were provided with a 12-hour light and dark environment, as well as food and water, and weighed every three days. The specific dosage is as follows:

[0197]

[0198]

[0199] 2. On day 22, the right ventricular systolic pressure (RVSP) of mice was measured by jugular vein catheterization. 1) Machine preparation: Select the mouse tidal volume on the small animal ventilator, then connect the computer to the BL-420N biological signal acquisition and processing system, select a single channel, exhaust the bubbles in the pipeline, and calibrate to zero; 2) Anesthesia: Weigh, anesthetize the mouse with 1% sodium pentobarbital solution (20 mg / kg) intraperitoneally, lay the mouse flat, check whether the depth of anesthesia is sufficient, and fix the mouse on the animal operating table after confirming that the mouse has no reflexes; 3) Disinfection: Disinfect the mouse neck and chest with medical alcohol; 4) Intubation: Use a scalpel to make a 1 cm straight incision from the base of the mouse neck. After separating the subcutaneous tissue, the trachea can be seen. Use ophthalmic scissors and tweezers to bluntly and sharply separate the soft tissue around the trachea, free the trachea, and use ophthalmic scissors to cut a "V"-shaped incision about 2 mm long. After inserting the trachea, it can be observed that the mouse chest rises and falls in line with the ventilator; 5) Pressure measurement: Use scissors to cut the chest skin to expose the subcutaneous tissue. Pay attention to blunt separation when cutting to avoid accidental injury. Then lift the xiphoid process with forceps, cut the chest with scissors, tear the pericardium with forceps to expose the heart. Insert the tube connected to the syringe into the right ventricle, and display the right ventricular pressure waveform on the computer for 10 seconds. Record and save the data.

[0200] 3. The mice were subsequently perfused by thoracotomy to obtain heart and lung tissues. After the right ventricular pressure was measured, saline containing sodium heparin was injected into the heart through the right ventricle to flush the heart and lung tissues, flushing the blood until the flushed liquid became clear and the lungs turned white. The fat tissue around the heart was cut off, the superior and inferior vena cava, aorta, pulmonary artery trunk and pulmonary vein were cut off, the left and right heart tissues were extracted and weighed, the left and right lung tissues were extracted, and three portions of the lung tissue were taken. One portion was fixed with paraformaldehyde and sent to the company for sectioning and HE staining, and the other two portions were frozen separately. Two portions were retained from the right ventricle, one portion was fixed with paraformaldehyde and sent to the company for sectioning and hematoxylin-eosin H&E and wheat germ agglutinin (WGA) staining, and the other portion was frozen for later use.

[0201] 4. Results: The animal experiment results showed that compared with the model group, the high-dose group of the nitrogen-four-membered ring-linked pyrazolopyridine PDE10A inhibitor A11 of the present invention can significantly alleviate the weight loss of rats after modeling (see Figure 4 ), reduce right heart systolic pressure RVSP (see Figure 5 A) and right heart hypertrophy index (see Figure 5 B), improve the intimal hyperplasia of the pulmonary arteriolar wall caused by pulmonary hypertension (see Figure 6 A in the figure), and the efficacy was close to that of the positive control tadalafil at the same dose. WGA staining of the right ventricle showed that the high-dose A11 group could significantly alleviate the hypertrophy of myocardial cells in rats after modeling, and the effect was similar to that of tadalafil (see Figure 6B). HE staining of lung tissue showed that the high-dose A11 group could significantly inhibit the thickening of vascular walls induced by SU5416+ hypoxia in rats (see Figure 7 ).

[0202] In summary, the pyrazolopyridine PDE10A inhibitor connected by a nitrogen tetra-membered ring of the present invention can be used as a drug molecule for treating pulmonary arterial hypertension.

[0203] The technical solution of the present invention is not limited to the above-mentioned specific embodiments. All technical variations made according to the technical solution of the present invention fall within the protection scope of the present invention.

Claims

1. Use of a pyrazolopyridine compound connected by a nitrogen four-membered ring in the preparation of a drug for treating pulmonary hypertension, characterized in that: The pyrazolopyridine compound connected by the nitrogen four-membered ring has a structure shown in formula (I): 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; Quinoxaline R6, R7 substituted quinoline and R6, R 10 Substituted Quinazolines One of; R4 is One of; R5 is H, C 1-5 Alkyl, isopropyl, -C (1-3) CH2OH, -C (1-3) One of CH2OCH3, -C(=O)CH3, -SO2CH3, -C(=O)OCH3; R6 is independently a mono- or di-substituted F atom, 18 One of F atom, Cl atom, hydroxyl group, methyl group, trifluoromethyl group, methoxy group and cyano group; R7 is independently hydroxy, methyl, trifluoromethyl, C 1-3 One of alkoxy, R4, -C(=O)-R4, R8, -CH2-R9, -C(=O)CH2-R9, -NHC(=O)-CH2-R9; R8 is: One of; R9 is One of; R 10 are independently methyl, ethyl, isopropyl, C 1-3 One of alkoxy, difluoromethoxy, difluoroethoxy, trifluoromethoxy, cyano, R8 and R9.

2. A drug for treating pulmonary hypertension, characterized in that: A pyrazolopyridine compound connected by an aza four-membered ring as claimed in claim 1.

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