Application of azetidine ring-linked pyrazolopyridine compound in preparation of drug for treating pulmonary arterial hypertension

Abstract

The present invention relates to the technical field of pharmaceuticals, and specifically discloses an application of an azetidine ring-linked pyrazolopyridine compound in the preparation of a drug for treating pulmonary arterial hypertension. Compared with the prior art, the azetidine ring-linked pyrazolopyridine PDE10A inhibitor of the present invention is capable of inhibiting enzymatic activity of PDE10A, thereby exerting a potential anti-pulmonary arterial hypertension effect. Therefore, the present invention also provides a new use of the compound, that is, the use of the azetidine ring-linked pyrazolopyridine PDE10A inhibitor in the preparation of a drug for treating pulmonary arterial hypertension.

Description

Application of pyrazolopyridine compounds with aza-4-membered rings in the preparation of drugs for treating pulmonary hypertension Technical Field

[0001] This invention relates to the field of pharmaceutical technology, particularly to the preparation of pyrazolopyridine compounds linked by a nitrogen-containing four-membered rings, and their application as drug molecules in the treatment of pulmonary hypertension. Background Technology

[0002] Pulmonary arterial hypertension (PAH) is a malignant pulmonary vascular disease with high morbidity and mortality. Pathologically, it is characterized by excessively high pulmonary artery pressure exceeding a certain threshold. Clinical symptoms include dyspnea, fatigue, weakness, decreased exercise tolerance, syncope, and chest pain. The pathogenesis of PAH is highly complex. Currently, many clinically available drugs regulate and intervene in intracellular cAMP and cGMP levels to exert therapeutic effects. Examples include phosphodiesterase (PDE5A) inhibitors (sildenafil, tadalafil), soluble guanylate cyclase (sGC) stimulators (riociguat) which exert their effects by regulating the intracellular NO / sGC / cGMP signaling pathway; and prostacyclin PGI2 analogs (eprostol) or prostacyclin PGI2 receptor agonists (selexipag) which exert their effects by regulating the intracellular PGI2 / cAMP signaling pathway. Theoretically, inhibiting intracellular PDE10A enzyme activity could, to some extent, regulate cAMP and cGMP levels, potentially exerting an anti-PAH effect.

[0003] Currently, reports of small-molecule selective inhibitors targeting PDE10A for the treatment of PAH are extremely rare; instead, they are almost exclusively focused on the treatment of central nervous system diseases. This necessitates that these PDE10A inhibitors possess good blood-brain barrier (BBB) ​​permeability to accumulate in the brain and maintain a certain therapeutic concentration. However, the reported use of these PDE10A inhibitors for peripheral tissue diseases (such as PAH) can lead to their accumulation in brain tissue due to their good BBB permeability, resulting in a strong inhibitory effect on the highly expressed PDE10A in brain tissue, thus significantly increasing the probability of adverse clinical events. On the other hand, the widely used selective PDE5A inhibitor sildenafil is rapidly metabolized in vivo and has poor selectivity for PDE6 (~16-fold), easily causing visual disturbances. Another selective PDE5A inhibitor, tadalafil, also has poor selectivity for PDE11 (~25-fold), easily causing adverse reactions such as muscle pain. Highly selective PDE10A inhibitors have a greater advantage in selectivity for the PDE family, typically exceeding 100-fold. Therefore, developing highly selective and active PDE10A inhibitors with low BBB cross-linking for the treatment of pulmonary arterial hypertension (PAH) is of greater clinical development value and is more innovative. Since PAH patients require lifelong medication, developing orally administered, highly selective PDE10A inhibitors with good drug-like properties is another major challenge. Summary of the Invention

[0004] To address the aforementioned technical problems, this invention provides the application of a nitrogen-containing four-membered ring-linked pyrazolopyridine compound in the preparation of a drug for treating pulmonary hypertension.

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

[0006] One object of this invention is to provide the use of aza-4-membered ring-linked pyrazolopyridine compounds in the preparation of medicaments for treating pulmonary hypertension, wherein the aza-4-membered ring-linked pyrazolopyridine compounds have the structure shown in formula (I):

[0007] Wherein: R1, R2, and R3 substituents are each independently H, halogen, or... 18 F, C 1-3 Alkyl, C 1-3 One of alkoxy, difluoromethoxy, difluoroethoxy, trifluoromethyl, trifluoromethoxy, acetyl, and cyano;

[0008] Quinoxaline R6- or R7-substituted quinolines and R6, R 10 Substituted quinazoline One of them;

[0009] R4 is One of them;

[0010] R5 represents H and C. 1-5 Alkyl, isopropyl, -C (1-3) CH2OH, -C (1-3) One of CH2OCH3, -C(=O)CH3, -SO2CH3, and -C(=O)OCH3;

[0011] R6 is an F atom that is either monosubstituted or disubstituted. 18 One of the following: F atom, Cl atom, hydroxyl group, methyl group, trifluoromethyl group, methoxy group, and cyano group;

[0012] R7 can be independently hydroxyl, methyl, trifluoromethyl, or C. 1-3 One of the following: alkoxy group, R4, -C(=O)-R4, R8, -CH2-R9, -C(=O)CH2-R9, -NHC(=O)-CH2-R9;

[0013] R8 is: One of them;

[0014] R9 is One of them;

[0015] R 10 Independently methyl, ethyl, isopropyl, C 1-3 One of alkoxy, difluoromethoxy, difluoroethoxy, trifluoromethoxy, cyano, R8, and R9.

[0016] A second objective of this invention is to provide a drug for treating pulmonary hypertension, comprising the aforementioned pyrazolopyridine compound linked by a nitrogen-containing four-membered ring.

[0017] Compared with the prior art, the pyrazolopyridine PDE10A inhibitor linked by the aza-four-membered ring 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 for this compound, namely, the application of the pyrazolopyridine PDE10A inhibitor linked by the aza-four-membered ring in the preparation of a drug for treating pulmonary hypertension. Attached Figure Description

[0018] Figure 1 shows the mean pulmonary artery pressure (A) and right ventricular systolic pressure (B) of rats in Example 2.

[0019] Figure 2 shows the ratio of pulmonary arterial wall thickness to cross-sectional diameter (%) in rats in Example 2 (A) and the measurement value of right ventricular hypertrophy index in rats (B).

[0020] Figure 3 shows a pathological tissue section of the rat pulmonary arteriole in Example 2. The upper part is a representative hematoxylin and eosin (H&E) staining image, the middle part is a representative Masson staining image, and the lower part is a representative wheat germ agglutinin (WGA) staining image of the right ventricle cross section.

[0021] Figure 4 shows the statistics of mouse weight detection in Example 3.

[0022] Figure 5 shows the measured values ​​of right ventricular systolic pressure (A) and right ventricular hypertrophy index (B) in mice in Example 3.

[0023] Figure 6 shows the ratio (%) of the wall thickness to the cross-sectional diameter of the pulmonary arteriole in mice in Example 3 and the individual area statistics of cardiomyocytes after WGA staining of the right ventricular cross section (B).

[0024] Figure 7 shows a pathological tissue section of the pulmonary arteriole in mice in Example 3. The upper part is a representative WGA staining image of the right ventricle transverse section, and the lower part is a representative hematoxylin and eosin (H&E) staining image.

[0025] Figure 8 shows the synthetic routes of compounds A1 to A12.

[0026] Figure 9 shows the synthetic routes of compounds M5 to M11.

[0027] Figure 10 shows the synthetic route of compounds A13 to A20.

[0028] Figure 11 shows the synthetic routes of compounds B1 to B6.

[0029] Figure 12 shows the synthetic route of compounds B7 to B22.

[0030] Figure 13 shows the synthetic route of compound B23.

[0031] Figure 14 shows the synthetic route of compounds B24 to B26.

[0032] Figure 15 shows the synthetic route of compound B27.

[0033] Figure 16 shows the synthetic route of compounds C1 to C2.

[0034] Figure 17 shows the synthetic route of compound C3.

[0035] Figure 18 shows the synthetic routes for compounds C4 to C8.

[0036] Figure 19 shows the synthetic route of compounds C9 to C10. Detailed Implementation

[0037] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to embodiments. The specific embodiments described herein are for illustrative purposes only and are not intended to limit the invention.

[0038] Example 1

[0039] This embodiment exemplifies some representative pyrazolopyridine PDE10A inhibitors with a nitrogen-containing four-membered ring linkage, the specific structures of which are as follows:

[0040] In this embodiment, the synthetic routes of compounds A1 to A12 are shown in Figure 8:

[0041] The synthesis of intermediate M1 was as follows: In a 250 mL round-bottom flask, 1-Boc-azacyclobutane-3-carboxylic acid (10 g, 49.7 mmol), cyclo(isopropyl)malonate (10.74 g, 74.5 mmol), and 4-dimethylaminopyridine (9.10 g, 1.5 mmol) were dissolved in 100 mL of dichloromethane. Under argon protection, a solution of N,N'-carbonyldiimidazole (9.67 g, 59.6 mmol) in 60 mL of dichloromethane was slowly added dropwise to the above system at 0 °C. The reaction mixture was allowed to rise naturally to room temperature and stirred overnight. The reaction was monitored by TLC. After the reaction was complete, a saturated citric acid aqueous solution was slowly added to the reaction system to adjust the pH to 5–6. Saturated brine (60 mL) was added, and the organic phase was separated. The aqueous phase was extracted twice with dichloromethane (30 mL). The organic phase was separated, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated by vacuum distillation to obtain a yellow oily compound. The compound was dissolved in anhydrous ethanol (60 mL), heated to 80 °C, and stirred overnight. The reaction was monitored by TLC. After the reaction was complete, the reaction mixture was concentrated by vacuum distillation. The crude product was purified by silica gel column chromatography to give a pale 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).

[0042] The synthesis of intermediate M2a was as follows: In a 125 mL round-bottom flask, intermediate M1 (3.02 g, 11.13 mmol) was dissolved in acetonitrile (60 mL), and 2-aminopyridine (3.15 g, 33.39 mmol) and carbon tetrabromide (7.38 g, 22.26 mmol) were added. The reaction mixture was stirred overnight at 80 °C, and the reaction was monitored by TLC. After the reaction was complete, the crude product was concentrated by vacuum distillation. A saturated aqueous solution of citric acid (30 mL) was added, and the mixture was extracted twice with ethyl acetate (30 mL x 2). The organic phase was collected, washed with saturated brine, and purified by silica gel column chromatography after vacuum distillation to obtain a yellow oily intermediate compound M2a (2.58 g, 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).

[0043] The synthesis of intermediate M3a was as follows: Intermediate M2a (2.58 g, 7.47 mmol) was dissolved in dichloromethane (40 mL) in a 125 mL round-bottom flask, 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 complete, the mixture was concentrated by vacuum distillation to give a yellow oily intermediate compound M3a (1.76 g, 96% yield). 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).

[0044] The synthesis of compound A1 was as follows: In a 50 mL round-bottom flask, intermediate M3a (100 mg, 0.41 mmol) and 2-chloroquinoline (80 mg, 0.49 mmol) were dissolved in anhydrous DMF (10 mL), and cesium carbonate (267 mg, 0.82 mmol) was added. The reaction mixture was heated to 110 °C overnight, and the reaction was monitored by TLC. After the reaction was complete, the cesium carbonate was filtered off, the filtrate was concentrated by vacuum distillation, and the crude product was purified by silica gel column chromatography to give a white solid compound A1 (72 mg, yield 62%). 1 H NMR (500MHz, CDCl3) δ9.34(d,J=7.0Hz,1H),7.86(d,J=9.0Hz,1H),7.74(d,J=8.5Hz,1H),7. 68(d,J=9.0Hz,1H),7.58(d,J=8.0Hz,1H),7.52(t,J=7.5Hz,1H),7.39(t,J=7.5Hz,1H),7.20 (t,J=7.5Hz,1H),7.00(t,J=7.0Hz,1H),6.66(d,J=9.0Hz,1H),4.69(dd,J=14.5,7.0Hz,1H), 4.63(t,J=7.5Hz,4H),4.45(q,J=7.0Hz,2H),1.47(t,J=7.0Hz,3H),HRMS(ESI-TOF)m / z[M+H] + calcd for C 22 H 20 N4O2393.1659, found 373.1668.

[0045] In this embodiment, the synthetic route of intermediates M5 to M11 (see Figure 9) is as follows:

[0046] The synthesis of intermediate M5 was as follows: 2-quinolinone-4-carboxylic acid (9.45 g, 50.0 mmol) was added to a 125 mL round-bottom flask, followed by careful addition of phosphorus oxychloride (30 mL). The reaction mixture was heated to 110 °C and stirred overnight, with the reaction monitored by TLC. After the reaction was complete, phosphorus oxychloride was removed by vacuum distillation to give a yellow oily intermediate compound M5 (100% yield).

[0047] The synthesis of intermediate M6 was as follows: Dry methanol (40 mL) was slowly added dropwise to cooled intermediate M5 (50.0 mmol) at 0 °C. The reaction mixture was then stirred at room temperature for 2 hours, and the reaction was monitored by TLC. After the reaction was complete, saturated sodium bicarbonate solution was added to the reaction mixture at 0 °C to adjust the pH to 9–10. The aqueous phase was extracted twice with ethyl acetate, and the organic phase was dried over anhydrous sodium sulfate. The filtrate was concentrated by vacuum distillation, and the crude product was purified by silica gel column chromatography to give a white solid intermediate compound M6 (5.87 g, 53% yield in two steps). 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).

[0048] The synthesis of intermediate 2-chloro-N-methylquinoline-4-carboxamide M7 was performed as follows: Cooled intermediate M5 (50.0 mmol) was dissolved in dry dichloromethane (40 mL), and methylamine hydrochloride (4.05 g, 60.0 mmol) and triethylamine (15.2 g, 150.0 mmol) were added. The reaction mixture was stirred at room temperature for 3 hours, and the reaction was monitored by TLC. After the reaction was complete, saturated sodium bicarbonate solution was added to the reaction mixture at 0 °C to adjust the pH to 9–10. The mixture was extracted twice with dichloromethane, and the organic phase was dried over anhydrous sodium sulfate. The filtrate was filtered and concentrated by vacuum distillation. The crude product was purified by silica gel column chromatography to give a white solid intermediate compound M7 (8.62 g, 78% yield in two steps). 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).

[0049] The synthesis of intermediate M8 was carried out as follows: using intermediate M5 (1.0 mmol) and 2,2-difluoroethylamine (97.2 mg, 1.2 mmol) as raw materials, and referring to the synthesis method of intermediate M7, a grayish-white solid intermediate compound M8 (60 mg, yield 48%) was obtained. 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).

[0050] The synthesis of intermediate M9 was carried out as follows: using intermediate M5 (1.0 mmol) and N-methylpiperazine (120 mg, 1.2 mmol) as raw materials, and referring to the synthesis method of intermediate M7, a pale yellow solid intermediate compound M9 (60 mg, yield 48%) was obtained. 1 H NMR(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).

[0051] The synthesis of intermediate M10 was carried out 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, and referring to the synthesis method of intermediate M7, a pale 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).

[0052] The synthesis of intermediate M11 was carried out as follows: using intermediate M5 (1.0 mmol) and N,N-diethylethylenediamine (139 mg, 1.2 mmol) as raw materials, and following the synthesis method of intermediate M7, a pale 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).

[0053] The synthesis of compound A2 was carried out as follows: using intermediate M3a (100 mg, 0.41 mmol) and intermediate M6 (108 mg, 0.49 mmol) as raw materials, the synthesis method of compound A1 was followed to obtain white solid compound A2 (121 mg, yield 69%). 1 H NMR (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=8.0Hz ,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] + calcd for C 24 H 22 N4O4431.1714, found 431.1715.

[0054] The synthesis of compound A3 was carried out as follows: using intermediate M3a (1.76 g, 7.18 mmol) and intermediate M7 (1.58 g, 7.18 mmol) as raw materials, the synthesis method of compound A1 was followed to obtain a pale yellow solid compound A3 (1.97 g, yield 64%). 1H NMR (400MHz, CDCl3) δ9.34(d,J=6.8Hz,1H),7.88(d,J=8.4Hz,1H),7.75(brs,1H ),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] + calcd for C 24 H 23 N5O3430.1874, found 430.1888.

[0055] The synthesis of intermediate M4 was as follows: In a 125 mL round-bottom flask, intermediate A3a (1.97 g, 4.59 mmol) was dissolved in methanol / water (30 mL / 10 mL), and sodium hydroxide (550 mg, 13.76 mmol) was added. The reaction mixture was heated to 65 °C and stirred for 3 hours, and the reaction was monitored by TLC. After the reaction was complete, methanol was removed by vacuum distillation to obtain an aqueous solution of the crude product. The pH of the system was adjusted to 4–5 with 5 M hydrochloric acid, and a solid precipitated. The solid was filtered under reduced pressure, the filter cake was washed with water, and dried in air to obtain a white solid intermediate compound M4 (1.51 g, 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).

[0056] The synthesis of compound A4 was as follows: In a 50 mL round-bottom flask, intermediate M4 (100 mg, 0.25 mmol) was dissolved in anhydrous N,N-dimethylformamide (10 mL). Then, 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) were added sequentially. The reaction mixture was stirred overnight at room temperature, and the reaction was monitored by TLC. After the reaction was complete, saturated brine was added, and the mixture was extracted with ethyl acetate. The organic phase was distilled under reduced pressure to obtain a crude product, which was purified by silica gel column chromatography to give a pale yellow solid compound A4 (87 mg, 75% yield). 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] + calcd for C 24 H 21 F2N5O3466.1685,found466.1690.

[0057] The synthesis of compound A5 was carried out as follows: using intermediate M4 (100 mg, 0.25 mmol) and isopropanol (30 mg, 0.5 mmol) as raw materials, and following the synthesis method of compound A4, a pale yellow compound A5 (66 mg, yield 60%) was obtained. 1H NMR (500MHz, CDCl3) δ9.34(d,J=7.0Hz,1H),7.80(d,J=8.0Hz,1H),7.68(d,J=9.0Hz,1 H),7.63(d,J=8.0Hz,1H),7.48(t,J=7.5Hz,1H),7.41(t,J=7.5Hz,1H),7.13(t,J=7.5 Hz,1H),7.02(t,J=7.0Hz,1H),6.39(s,1H),5.41–5.30(m,1H),4.64–4.55(m,1H),4.5 3–4.40(m,4H),3.11(d,J=4.5Hz,3H),1.46(d,J=6.0Hz,6H).HRMS(ESI-TOF)m / z:[M+H] + calcd for C 25 H 25 N5O3444.2030,found444.2032.

[0058] The synthesis of compound A6 was carried out as follows: using intermediate M4 (100 mg, 0.25 mmol) and oxetane-3-ol (37 mg, 0.5 mmol) as raw materials, and following the synthesis method of compound A4, a pale yellow compound A6 (77 mg, yield 68%) was obtained. 1 H NMR (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] + calcd for C 25 H 23 N5O4458.1823,found 458.1826.

[0059] The synthesis of compound A7 was carried out as follows: using intermediate M4 (100 mg, 0.25 mmol) and tetrahydropyran-4-ol (51 mg, 0.5 mmol) as raw materials, and following the synthesis method of compound A4, a pale yellow compound A7 (83 mg, yield 69%) was obtained.

[0060] 1 H NMR(500MHz, CDCl3)δ9.32(d,J=6.0Hz,1H),7.82(d,J=8.0Hz,1H),7.77–7.61(m,2H),7.50(t, J=7.5Hz,1H),7.44(t,J=7.5Hz,1H),7.15(t,J=6.5Hz,1H),7.06(t,J=6.0Hz,1H),6.44(s,1H) ,5.28(s,1H),4.66–4.58(m,1H),4.57–4.41(m,4H),4.13–4.01(m,2H),3.64(t,J=10.0Hz,2H) ,3.11(d,J=3.0Hz,3H),2.18–2.06(m,2H),1.98–1.81(m,2H).HRMS(ESI-TOF)m / z:[M+H]+calcd for C 27 H 27 N5O4486.2136,found 486.2135.

[0061] The synthesis of compound A8 was carried out as follows: using intermediate M3a (100 mg, 0.41 mmol) and intermediate M8 (133 mg, 0.49 mmol) as raw materials, the synthesis method of compound A1 was followed to obtain a pale yellow solid compound A8 (98 mg, yield 50%). 1 H NMR (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]+ calcd for C 25 H 23 F2N5O3480.1842,found480.1845.

[0062] The synthesis of compound A9 was carried out as follows: using intermediate M3a (100 mg, 0.41 mmol) and intermediate M9 (142 mg, 0.49 mmol) as raw materials, the synthesis method of compound A2 was followed to obtain a pale yellow solid compound A9 (134 mg, yield 66%). 1 H NMR (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] + calcd for C 28 H 30 N6O3499.2452, found499.2447.

[0063] The synthesis of compound A10 was carried out as follows: intermediate M3a (100 mg, 0.41 mmol) and intermediate M10 (196 mg, 0.49 mmol) were used as raw materials, and the synthesis method of compound A2 was followed. The protecting group was removed by trifluoroacetic acid to obtain a pale yellow solid compound A10 (135 mg, 65% yield in 2 steps). 1H NMR (500MHz, CDCl3) δ9.35(d,J=7.0Hz,1H),7.75(dd,J=11.5,9.0Hz,1H),7.70(d,J=9.0Hz,1H),7.64–7.52(m ,2H),7.42(t,J=8.0Hz,1H),7.25(t,J=8.0Hz,1H),7.03(t,J=7.0Hz,1H),6.58(d,J=18.5Hz,1H),4.79–4.56(m ,5H),4.47(q,J=7.0Hz,2H),4.01–3.61(m,2H),3.23–3.14(m,1H),3.12–2.98(m,2H),2.80(t,J=4.5Hz,1H),1 .49(td,J=7.5,1.5Hz,3H),0.92–0.69(m,2H),0.60–0.48(m,1H),0.33–0.10(m,1H).HRMS(ESI-TOF)m / z:[M+H] + calcd for C 29 H 30 N6O3511.2452, found 511.2477.

[0064] The synthesis of compound A11 was as follows: In a 50 mL round-bottom flask, intermediates M11 (305 mg, 1.0 mmol) and M3a (245 mg, 1.0 mmol) were dissolved in anhydrous DMF (20 mL), and cesium carbonate (652 mg, 2.0 mmol) was added. The reaction mixture was heated to 110 °C and reacted overnight, with the reaction monitored by TLC. After the reaction was complete, the cesium carbonate was filtered off, and the resulting filtrate was concentrated by vacuum distillation. The crude product was purified by silica gel column chromatography to give a yellow solid compound A11 (272 mg, yield 53%). 1H NMR (400MHz, CDCl3) δ9.35(d,J=6.8Hz,1H),7.99(d,J=8.4Hz,1H),7.75(d,J=8.4Hz,1H),7.69(d,J=8.8H z,1H),7.54(td,J=7.2,1.2Hz,1H),7.42(td,J=7.2,1.2Hz,1H),7.23(t,J=8.0Hz,1H),7.03(td,J=6.8,1. 2Hz,1H),6.84–6.77(m,1H),6.76(s,1H),4.75–4.67(m,1H),4.65–4.59(m,4H),4.46(q,J=7.2Hz,2H),3.6 4–3.55(m,2H),2.71(t,J=6.0Hz,2H),2.58(q,J=7.2Hz,4H),1.48(t,J=7.2Hz,3H),1.03(t,J=7.2Hz,6H).

[0065] The synthesis of compound A12 was performed as follows: Following the synthetic method for compound A1, 2-chloroquinoline was reacted with intermediate M3b to yield a pale 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).

[0066] In this embodiment, the synthetic routes for compounds A13 to A20 (see Figure 10) are as follows:

[0067] The synthesis of compound A13 was performed as follows: In a 50 mL round-bottom flask, intermediate M4 (100 mg, 0.25 mmol) was dissolved in anhydrous N,N-dimethylformamide (10 mL), followed by the sequential addition of methylamine hydrochloride (20 mg, 0.30 mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (114 mg, 0.30 mmol), and diisopropylethylamine (97 mg, 0.75 mmol). The reaction mixture was stirred overnight at room temperature, and the reaction was monitored by TLC. After the reaction was complete, the mixture was concentrated by vacuum distillation, and the crude product was purified by silica gel column chromatography to give a 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] + calcd for C 23 H 22 N6O2415.1877,found 415.1875.

[0068] The synthesis of compound A14 was carried out as follows: using intermediate M4 (100 mg, 0.25 mmol) and 2-methoxyethylamine (23 mg, 0.30 mmol) as raw materials, and following the synthesis method of compound A13, a pale yellow solid compound A14 (54 mg, yield 47%) was obtained. 1H NMR(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] + calcd for C 25 H 26 N6O3459.2139,found 459.2139.

[0069] The synthesis of compound A15 was carried out as follows: using intermediate M4 (100 mg, 0.25 mmol) and 2,2-difluoroethylamine (24 mg, 0.30 mmol) as raw materials, and following the synthesis method of compound A13, a pale yellow solid compound A15 (66 mg, yield 57%) was obtained. 1 H NMR (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,1 H),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] + calcd for C 24 H 22 F2N6O2465.1845, found 465.1842.

[0070] The synthesis of compound A16 was carried out as follows: using intermediate M4 (100 mg, 0.25 mmol) and cyclopropylamine (17 mg, 0.30 mmol) as raw materials, and following the synthesis method of compound A13, a pale yellow solid compound A16 (66 mg, yield 60%) was obtained. 1 H NMR (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),2.96 –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] + calcd for C 25 H 24 N6O2441.2034,found441.2036.

[0071] The synthesis of compound A17 was carried out as follows: using intermediate M4 (100 mg, 0.25 mmol) and N,N-diethylethylenediamine (35 mg, 0.30 mmol) as raw materials, and following the synthesis method of compound A13, a pale 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]+ calcd for C 28 H 33 N7O2500.2768, found 500.2769.

[0072] The synthesis of compound A18 was carried out as follows: using intermediate M4 (100 mg, 0.25 mmol) and N,N-diethyl-N'-methylethylenediamine (39 mg, 0.30 mmol) as raw materials, and following the synthesis method of compound A13, a pale 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] + calcd for C 29 H 35 N7O2514.2925, found 514.2922.

[0073] The synthesis of intermediate M13 was as follows: In a 125 mL round-bottom flask, A11 (272 mg, 0.53 mmol) was dissolved in methanol / water (20 mL / 5 mL), and sodium hydroxide (64 mg, 1.59 mmol) was added. The reaction mixture was heated to 65 °C and stirred for 3 hours, and the reaction was monitored by TLC. After the reaction was complete, methanol was removed by vacuum distillation to obtain an aqueous solution of the crude product. The pH of the system was adjusted to 4–5 with 5 M hydrochloric acid, and a solid precipitated. The solid was filtered under reduced pressure, the filter cake was washed with water, and dried in air to obtain a pale yellow solid M12 (236 mg, yield 92%).

[0074] The synthesis of compound A19 was carried out as follows: using intermediate M12 (100 mg, 0.21 mmol) and dimethylamine hydrochloride (20 mg, 0.25 mmol) as raw materials, and following the synthesis method of compound A13, a pale yellow solid compound A19 (98 mg, yield 77%) was obtained. 1 H NMR (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] + calcd for C 29 H 35 N7O2514.2925, found 514.2944.

[0075] The synthesis of compound A20 was carried out as follows: using intermediate M12 (100 mg, 0.21 mmol) and methoxymethylamine (18 mg, 0.30 mmol) as raw materials, and following the synthesis method of compound A13, a pale yellow solid compound A20 (95 mg, yield 72%) was obtained. 1H NMR (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=9.2 Hz,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] + calcd for C 29 H 35 N7O3552.2694, found 552.2693.

[0076] In this embodiment, the synthetic routes of compounds B1 to B6 (see Figure 11) are as follows:

[0077] The synthesis of intermediates M14a-f was as follows: Intermediate M1 (1.08 mg, 4.0 mmol) was dissolved in acetonitrile (20 mL) in a 125 mL round-bottom flask. Various R3- or R4-substituted 2-aminopyridine P-2-A derivatives (6.0 mmol) and carbon tetrabromide (1.99 g, 6.0 mmol) were added. The reaction mixture was stirred overnight at 80 °C, and the reaction was monitored by TLC. After the reaction was complete, the mixture was concentrated under reduced pressure by distillation. The crude product was purified by silica gel column chromatography to obtain the yellow oily intermediate compounds M14a-f.

[0078] 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).

[0079] 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).

[0080] 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).

[0081] 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).

[0082] 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).

[0083] 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).

[0084] The synthesis of intermediates M15a-f was as follows: In a 50 mL round-bottom flask, intermediate 14a-f (1.0 mmol) was dissolved in methanol (10 mL) and water (2 mL), and sodium hydroxide (120 mg, 3.0 mmol) was added. The reaction mixture was stirred overnight at 60 °C, and the reaction was monitored by TLC. After the reaction was complete, the crude product was distilled under reduced pressure to obtain an aqueous solution. The pH was adjusted to 3-4 with saturated citric acid solution, and the mixture was extracted with dichloromethane. The organic phase was dried over anhydrous sodium sulfate and concentrated to obtain white or pale yellow solid intermediates M15a-f (yield 76-88%), which did not require purification and were used directly in the next reaction.

[0085] The synthesis of intermediates M16a-f was as follows: In a 50 mL round-bottom flask, intermediate compound M15a-f (0.5 mmol) was dissolved in anhydrous N,N-dimethylformamide (10 mL). Dimethylamine hydrochloride (DMA·HCl) (1.0 mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (228 mg, 0.6 mmol), and diisopropylethylamine (194 mg, 1.5 mmol) were added sequentially. The reaction mixture was stirred overnight at room temperature, and the reaction was monitored by TLC. After the reaction was complete, the crude product was concentrated by vacuum distillation. Saturated brine (30 mL) was added, and the mixture was extracted twice with ethyl acetate. The extract was dried over anhydrous sodium sulfate, filtered, and the crude product was concentrated by vacuum distillation. The crude product was purified by silica gel column chromatography to obtain pale yellow or white solid intermediate compounds M16a-f.

[0086] 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).

[0087] 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).

[0088] Intermediate M16c, pale 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).

[0089] 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).

[0090] Intermediate M16e, pale yellow solid, 91% yield. 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).

[0091] 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).

[0092] The synthesis of intermediates M17a-f was as follows: In a 50 mL round-bottom flask, compound M16a-f (0.5 mmol) was dissolved in methanol (5 mL), and concentrated hydrochloric acid (0.5 mL) was added. The reaction mixture was stirred overnight at room temperature, and the reaction was monitored by TLC. After the reaction was complete, the mixture was concentrated by vacuum distillation to obtain pale yellow solid intermediates M17a-f.

[0093] 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).

[0094] 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).

[0095] 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).

[0096] Intermediate M17d, yield 90%. 1 H NMR (500MHz, CD3OD) δ8.15(s,1H),7.57(d,J=9.0Hz,1H),7.36(dd,J=9.0,1.5Hz,1H),4.47–4.34(m,5H),3.09(s,6H),2.37(s,3H).

[0097] 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).

[0098] 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).

[0099] In this embodiment, the synthesis of compounds B1–B6 was performed as follows: Intermediates M17a-f (0.25 mmol) and M7 (66 mg, 0.30 mmol) were dissolved in anhydrous DMF (10 mL) in a 50 mL flask, and cesium carbonate (244 mg, 0.75 mmol) was added. The reaction mixture was heated to 110 °C and reacted overnight, with the reaction monitored by TLC. After the reaction was complete, the cesium carbonate was removed by filtration, and the filtrate was distilled under reduced pressure to obtain crude products. These crude products were purified by silica gel column chromatography to yield pale yellow or white solid compounds B1–B6, respectively.

[0100] Compound B1, 75 mg, pale 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] + calcd for C 24 H 24 N6O2429.2034, found429.2033.

[0101] 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] + calcd for C 25 H 26 N6O2443.2190, found 443.2189.

[0102] Compound B3, 50 mg, pale 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] + calcd for C 24 H 23 N6O2F 447.1939, found 447.1933.

[0103] 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] + calcd for C 25 H 26 N6O2443.2190, found443.2193.

[0104] Compound B5, 52 mg, pale 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] + calcd for C 24 H 23 N6O2F 469.1759, found 469.1770.

[0105] 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] + calcd for C 25 H 26 N6O3459.2139, found 459.2142.

[0106] In this embodiment, the synthetic routes for compounds B7 to B22 (see Figure 12) are as follows:

[0107] The synthesis of intermediate M18 was as follows: In a 125 mL round-bottom flask, intermediates M17e (2.0 g, 7.62 mmol) and M6 (2.03 g, 9.15 mmol) were dissolved in anhydrous DMF (30 mL), and cesium carbonate (4.96 g, 15.24 mmol) was added. The reaction mixture was heated to 110 °C overnight, and the reaction was monitored by TLC. After the reaction was complete, the cesium carbonate was removed by filtration, and the filtrate was distilled under reduced pressure to obtain the crude product, which was purified by silica gel column chromatography to give 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.

[0108] The synthesis of intermediate M19 was as follows: In a 125 mL round-bottom flask, intermediate M18 (1.64 g, 3.66 mmol) was dissolved in methanol (30 mL) and water (6 mL), and sodium hydroxide (440 mg, 11.0 mmol) was added. The reaction mixture was stirred overnight at 60 °C, and the reaction was monitored by TLC. After the reaction was complete, the crude product was distilled under reduced pressure to obtain an aqueous solution. The pH was adjusted to 3-4 with the addition of saturated citric acid solution, and a solid precipitated. The solid was filtered, the filter cake was washed with water, and dried in air to obtain a white solid compound M19 (1.46 g, 92% yield).

[0109] The synthesis of compounds B7, B8, B10, B11, and B15 was as follows: In a 50 mL flask, compound M19 (100 mg, 0.23 mmol) was dissolved in anhydrous N,N-dimethylformamide (10 mL). The corresponding amine compounds (0.28 mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (105 mg, 0.28 mmol), and diisopropylethylamine (89 mg, 0.69 mmol) were added sequentially. The reaction mixture was stirred overnight at room temperature, and the reaction was monitored by TLC. After the reaction was complete, the crude product was concentrated by vacuum distillation. Saturated brine (30 mL) was added, and the mixture was extracted twice with ethyl acetate. The extract was dried over anhydrous sodium sulfate, filtered, and concentrated by vacuum distillation. The crude product was purified by silica gel column chromatography to obtain pale yellow or white solid compounds B7, B8, B10, B11, and B15, respectively.

[0110] Compound B7, 72 mg, pale 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] + calcd for C 29 H 34 N7O2F 532.2831, found 532.2860.

[0111] Compound B8, 62 mg, pale 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] + calcd for C 27 H 30 N7O2F 504.2518, found 504.2544.

[0112] Compound B10, 51 mg, pale 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] + calcd for C 25 H 24 N7O2FS 506.1769,found 506.1770.

[0113] Compound B11, 52 mg, pale 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] + calcd for C 26 H 22 N7O3FS2564.1282, found 564.1307.

[0114] Compound B15 is a pale yellow solid with a yield of 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] + calcd for C 30 H 32 N7O2F 542.2674,found 542.2680.

[0115] In a 50 mL round-bottom flask, compound M19 (100 mg, 0.23 mmol) was dissolved in anhydrous N,N-dimethylformamide (10 mL). The corresponding amine compounds (0.28 mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (105 mg, 0.28 mmol), and diisopropylethylamine (89 mg, 0.69 mmol) were added sequentially. The reaction mixture was stirred overnight at room temperature, and the reaction was monitored by TLC. After the reaction was complete, the system was concentrated under reduced pressure to obtain a crude product. Saturated brine (30 mL) was added, and the mixture was extracted twice with ethyl acetate, dried over anhydrous sodium sulfate, and the filtrate was filtered. The crude product was concentrated under reduced pressure and purified by silica gel column chromatography to give a pale yellow or white solid compound. The obtained compound was dissolved in dichloromethane (20 mL), and trifluoroacetic acid (2 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 crude product was obtained by vacuum distillation. The pH was adjusted to 9-10 by adding saturated sodium bicarbonate solution. The product was extracted twice with ethyl acetate (10 mL). The organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and the filtrate was evaporated under vacuum. The crude product was purified by silica gel column chromatography to give pale yellow solids B9, B12-B14, B16 and B18.

[0116] 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] + calcd for C 26 H 28 N7O2F 490.2361,found490.2356.

[0117] Compound B12, 57 mg, pale 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] + calcd for C 27 H 28 N7O2F 502.2361, found 502.2358.

[0118] Compound B13, 52 mg, pale 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 N7O2F 516.2518, found 516.2495。

[0119] Compound B14, 54 mg, light 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。

[0120] Compound B16, 50 mg, pale 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] + calcd for C 29 H 30 N7O2F 550.2337, found 550.2358.

[0121] Compound B18, 45 mg, pale 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] + calcd for C 28 H 30 N7O2F516.2518, found 516.2505.

[0122] The synthesis of compound B17 was as follows: In a 50 mL round-bottom flask, compound B16 (100 mg, 0.19 mmol) was dissolved in methanol (6 mL), followed by the addition of formaldehyde aqueous solution (0.2 mL, 37%) and sodium cyanoborohydride (36 mg, 0.57 mmol). The reaction mixture was stirred overnight at room temperature, and the reaction was monitored by TLC. After the reaction was complete, the crude product was concentrated by vacuum distillation. Saturated brine (10 mL) was added, and the mixture was extracted twice with ethyl acetate. The extract was dried over anhydrous sodium sulfate, and the filtrate was concentrated by vacuum distillation. The crude product was purified by silica gel column chromatography to give a pale yellow solid compound B17 (82 mg, 80% yield). 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] + calcd for C 30 H 32 N7O2F 542.2674,found 542.2669.

[0123] The synthesis of compound B19 was as follows: using compound B12 (80 mg, 0.16 mmol) as the starting material, and following the synthesis method of compound B17, a pale 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).

[0124] The synthesis of compound B20 was as follows: using compound B18 (100 mg, 0.19 mmol) as the starting material, and following the synthesis method of compound B17, a pale 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).

[0125] The synthesis of compound B21 was as follows: using compound B13 (120 mg, 0.23 mmol) as the starting material, and following the synthesis method of compound B17, a pale 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).

[0126] The synthesis of compound B22 was carried out as follows: using compound B14 (120 mg, 0.23 mmol) as the starting material, and following the synthesis method of compound B17, a pale 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).

[0127] In this embodiment, the synthetic route of compound B23 (see Figure 13) is as follows:

[0128] The synthesis of intermediate M20 was as follows: In a 125 mL round-bottom flask, 2-(1-tert-butoxycarbonyl)azacyclobutane-3-yl)-6-fluoroimidazolo[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), and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (4.18 g, 11.0 mmol) were dissolved in N,N-dimethylformamide (30 mL). The reaction mixture was stirred overnight at room temperature, and the reaction was monitored by TLC. After the reaction was complete, the filtrate was concentrated by vacuum distillation, and the crude product was purified by silica gel column chromatography to give 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).

[0129] The synthesis of intermediate M21 was as follows: M20 (1.82 g, 4.82 mmol) and methylmagnesium bromide (5 mL, 14.44 mmol) were dissolved in tetrahydrofuran (40 mL) in a 125 mL round-bottom flask at 0 °C. The reaction mixture was then stirred overnight at room temperature, and the reaction was monitored by TLC. After the reaction was complete, a saturated ammonium chloride solution was added to the reaction mixture at 0 °C. The aqueous phase was extracted twice with ethyl acetate, and the organic phase was dried over anhydrous sodium sulfate. The filtrate was concentrated by vacuum distillation, and the crude product was purified by silica gel column chromatography to give 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).

[0130] The synthesis of intermediate M22 was as follows: In a 50 mL round-bottom flask, compound M21 (200 mg, 0.6 mmol) was dissolved in methanol (10 mL), and concentrated hydrochloric acid (1.0 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 mixture was concentrated by vacuum distillation to give a white solid intermediate compound M22 (140 mg, 100% yield). 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).

[0131] The synthesis of compound B23 was as follows: In a 50 mL round-bottom flask, intermediates M22 (100 mg, 0.43 mmol) and M7 (95 mg, 0.43 mmol) were dissolved in anhydrous DMF (10 mL), and cesium carbonate (420 mg, 1.29 mmol) was added. The reaction mixture was heated to 110 °C and reacted overnight, with the reaction monitored by TLC. After the reaction was complete, the cesium carbonate was removed by filtration, and the filtrate was distilled under reduced pressure to obtain the crude product, which was purified by silica gel column chromatography to give 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).

[0132] In this embodiment, the synthetic routes for compounds B24 to B26 (see Figure 14) are as follows:

[0133] The synthesis of intermediate M23 was as follows: In a 100 mL round-bottom flask, ethyl 2-(azacyclobutan-3-yl)-6-fluoroimidazolo[1,2-a]pyridine-3-carboxylate M14e (400 mg, 1.52 mmol) and 2-chloro-4-methylquinoline were 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). The reaction mixture was heated at 100 °C overnight under argon protection. After the reaction was complete, the mixture was distilled under reduced pressure to obtain a crude product, which was purified by silica gel column chromatography to give a pale yellow solid intermediate compound M23 (128 mg, yield 54%). 1 H NMR(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).

[0134] The synthesis of intermediate M24 was as follows: In a 50 mL round-bottom flask, intermediate M23 (295 mg, 0.73 mmol) was dissolved in methanol (10 mL) and water (3 mL), and sodium hydroxide (88 mg, 2.19 mmol) was added. The reaction mixture was stirred overnight at 60 °C, and the reaction was monitored by TLC. After the reaction was complete, the crude product was distilled under reduced pressure to obtain an aqueous solution. The pH was adjusted to 3-4 with 5 M hydrochloric acid solution, and a solid precipitated. The solid was filtered, the filter cake was washed with water, and dried in air to obtain a pale yellow solid intermediate compound M24 (240 mg, yield 88%).

[0135] The synthesis of intermediate M25 was as follows: In a 125 mL round-bottom flask, intermediate M24 (200 mg, 0.53 mmol), 3,6-diazabicyclo[3.1.1]heptane-6-carboxylic acid tert-butyl ester (126 mg, 0.64 mmol), N,N-diisopropylethylamine (205 mg, 1.59 mmol), and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (242 mg, 0.64 mmol) were dissolved in N,N-dimethylformamide (10 mL). The reaction mixture was stirred overnight at room temperature, and the reaction was monitored by TLC. After the reaction was complete, the filtrate was concentrated by vacuum distillation, and the crude product was purified by silica gel column chromatography to give 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).

[0136] The synthesis of compound B24 was as follows: In a 50 mL round-bottom flask, 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 overnight at room temperature, and the reaction was monitored by TLC. After the reaction was complete, the mixture was distilled under reduced pressure to obtain a crude product, which was purified by silica gel column chromatography to give 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).

[0137] The synthesis of compound B25 was as follows: In a 50 mL round-bottom flask, compound B24 (143 mg, 0.31 mmol) was dissolved in methanol (10 mL), followed by the addition of formaldehyde aqueous solution (1 mL, 37%) and sodium cyanoborohydride (59 mg, 0.94 mmol). The reaction mixture was stirred overnight at room temperature, and the reaction was monitored by TLC. After the reaction was complete, the crude product was concentrated by vacuum distillation. Saturated brine (10 mL) was added, and the mixture was extracted twice with ethyl acetate. The extract was dried over anhydrous sodium sulfate, and the filtrate was concentrated by vacuum distillation. The crude product was purified by silica gel column chromatography to give compound B25 (66 mg, 45% yield) as a white solid. 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).

[0138] The synthesis of compound B26 was as follows: In a 50 mL flask, intermediate M24 (200 mg, 0.53 mmol) was dissolved in anhydrous N,N-dimethylformamide (10 mL). N-methylpiperazine (106 mg, 1.06 mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (242 mg, 0.64 mmol), and diisopropylethylamine (206 mg, 1.59 mmol) were added sequentially. The reaction mixture was stirred overnight at room temperature, and the reaction was monitored by TLC. After the reaction was complete, the crude product was concentrated by vacuum distillation. Saturated brine (30 mL) was added, and the mixture was extracted twice with ethyl acetate. The extract was dried over anhydrous sodium sulfate, filtered, and concentrated by vacuum distillation. The crude product was purified by silica gel column chromatography to give compound B26 (76 mg, 31% yield) as a pale yellow or white solid. 1 H NMR(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),6.4 4(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).

[0139] In this embodiment, the synthetic route of compound B27 (see Figure 15) is as follows:

[0140] The synthesis of intermediate M26 was as follows: 2,4-Dibromoquinoline (1.0 g, 3.48 mmol) was dissolved in anhydrous 1,4-dioxane (20 mL) in a 50 mL round-bottom flask. The mixture was heated to 90 °C, and 40% HBr (4 mL) was added dropwise. The reaction mixture was heated at 90 °C overnight. After the reaction was complete, the crude product was concentrated by vacuum distillation and purified by silica gel column chromatography to give a grayish-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).

[0141] The synthesis of intermediate M27 was as follows: In a 100 mL round-bottom flask, 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) were dissolved in 1,4-dioxane (20 mL) and H2O (5 mL). K2CO3 (977 mg, 3.0 mmol) and [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride Pd(dppf)Cl2 (73 mg, 0.10 mmol) were then added. The reaction mixture was heated to 90 °C overnight under argon protection. After the reaction was complete, the mixture was distilled under reduced pressure to obtain a crude product, which was purified by silica gel column chromatography to give a pale 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).

[0142] The synthesis of intermediate M28 was as follows: In a 100 mL round-bottom flask, intermediate M27 (575 mg, 2.39 mmol) was dissolved in toluene (20 mL), followed by the addition of phosphorus oxychloride (2 mL). The reaction mixture was heated to 100 °C for 4 hours. After the reaction was complete, the mixture was distilled under reduced pressure to obtain a crude product, which was purified by silica gel column chromatography to give a pale 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).

[0143] The synthesis of compound B27 was as follows: In a 100 mL round-bottom flask, intermediates M28 (570 mg, 2.20 mmol) and M17e (578 mg, 2.20 mmol) were 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). The reaction mixture was heated at 100 °C overnight under argon protection. After the reaction was complete, the mixture was distilled under reduced pressure to obtain a crude product, which was purified by silica gel column chromatography to give a pale 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).

[0144] In this embodiment, the synthetic routes for compounds C1 to C2 (see Figure 16) are as follows:

[0145] The synthesis of intermediate M29 was as follows: Methyl 2-amino-5-methylbenzoate (1.65 g, 10.0 mmol) was dissolved in acetic acid (10 mL) in a 125 mL round-bottom flask. An aqueous solution of potassium cyanate (973 mg, 12.0 mmol) (10 mL) was slowly added dropwise. The reaction mixture was heated to 60 °C overnight, and the reaction was monitored by TLC. After the reaction was complete, the mixture was cooled to room temperature, and water (40 mL) was added to the reaction mixture. The mixture was stirred for 10 minutes, and the precipitated solid was filtered off. The filter cake was washed with water and dried in 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).

[0146] The synthesis of intermediate M30 was as follows: In a 125 mL round-bottom flask, intermediate M29 (1.68 g, 8.1 mmol) was dissolved in anhydrous ethanol (30 mL), and sodium hydroxide (645 mg, 16.2 mmol) was added. The reaction mixture was heated to 80 °C and refluxed overnight, and the reaction was monitored by TLC. After the reaction was complete, the mixture was cooled to room temperature, and a solid precipitated out. The solid was filtered, the filter cake was washed with water, and allowed to dry naturally in air to give a white solid intermediate compound M30 (1.42 g, 100% yield). 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).

[0147] The synthesis of intermediate M31 was as follows: In a 125 mL round-bottom flask, intermediate M30 (1.42 g, 8.1 mmol) and phosphorus oxychloride (20 mL) were mixed. The reaction mixture was heated to reflux at 110 °C overnight, and the reaction was monitored by TLC. After the reaction was complete, the mixture was cooled to room temperature, and phosphorus oxychloride was removed by vacuum distillation to obtain the crude product. Dichloromethane and triethylamine were slowly added to the cooled crude product until the pH of the system was 8–9. The mixture was concentrated under reduced pressure and purified by silica gel column chromatography to obtain a white solid intermediate compound M31 (1.33 g, 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).

[0148] The synthesis of intermediate M32a was as follows: In a 125 mL round-bottom flask, intermediate M31 (891 mg, 4.18 mmol) was dissolved in anhydrous acetonitrile (40 mL), and 2,4-dimethoxybenzylamine (699 mg, 4.18 mmol) and diisopropylethylamine (1.62 g, 12.54 mmol) were added sequentially. The reaction mixture was heated at 50 °C overnight, and the reaction was monitored by TLC. After the reaction was complete, the mixture was cooled to room temperature, and a solid precipitated. The precipitated solid was filtered, the filter cake was washed with water, and dried in air to give a white solid intermediate compound M32a (1.32 g, 92% yield).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).

[0149] The synthesis of intermediate M32b was as follows: using intermediate M31 (426 mg, 2.0 mmol) and 3,8-diazabicyclo[3.2.1]octane-3-carboxylic acid tert-butyl ester (424 mg, 2.0 mmol) as raw materials, and 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).

[0150] The synthesis of intermediate M33a was as follows: In a 50 mL round-bottom flask, intermediates M32a (343 mg, 1.0 mmol) and M7 (262 mg, 1.0 mmol) were dissolved in anhydrous DMF (10 mL), and cesium carbonate (1.62 g, 3.0 mmol) was added. The reaction mixture was heated to 110 °C and reacted overnight, with the reaction monitored by TLC. After the reaction was complete, the cesium carbonate was removed by filtration, and the filtrate was distilled under reduced pressure to obtain the crude product, which was purified by silica gel column chromatography to give the pale yellow solid intermediate compound M33a (158 mg, 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).

[0151] The synthesis of intermediate M33b was as follows: using intermediates M32b (200 mg, 0.51 mmol) and M7 (135 mg, 0.51 mmol) as raw materials, and referring to the synthesis method of compound M33a, a pale 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).

[0152] The synthesis of compound C1 was as follows: In a 50 mL round-bottom flask, intermediate M33a (158 mg, 0.28 mmol) was dissolved in methanol (10 mL), and concentrated hydrochloric acid (1 mL) was carefully added. The reaction mixture was heated to 65 °C for 24 hours, and the reaction was monitored by TLC. After the reaction was complete, the mixture was distilled under reduced pressure to obtain a crude product, which was purified by silica gel column chromatography to give a pale 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] + calcd for C 22 H 22 N7OF 420.1943,found420.1946.

[0153] The synthesis of compound C2 was as follows: In a 50 mL round-bottom flask, 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 overnight at room temperature, and the reaction was monitored by TLC. After the reaction was complete, the mixture was distilled under reduced pressure to obtain a crude product, which was purified by silica gel column chromatography to give a pale 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] + calcd for C 28 H 31 N8OF 515.2678, found 515.2682.

[0154] In this embodiment, the synthetic route of compound C3 (see Figure 17) is as follows:

[0155] The synthesis of intermediate M34 was carried out as follows: using 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) as raw materials, and referring to the synthesis method of compound M32a, a pale yellow solid intermediate compound M34 (1.37 g, yield 73%) was 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).

[0156] The synthesis of intermediate M35 was carried out as follows: using intermediate M34 (187 mg, 0.50 mmol) and M17e (131 mg, 0.50 mmol) as raw materials, and referring to the synthesis method of compound M33a, a pale 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).

[0157] The synthesis of compound C3 was as follows: using intermediate M32 (130 mg, 0.22 mmol) as the starting material, and following the synthesis method of compound C2, a pale 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] + calcd for C 27 H 29 N8OF 501.2521, found 501.2518.

[0158] In this embodiment, the synthetic routes for compounds C4 to C8 (see Figure 18) are as follows:

[0159] The synthesis of intermediate M36 was as follows: using compound M14e (825 mg, 2.27 mmol) as the starting material, and following 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).

[0160] The synthesis of compound C4 was carried out as follows: using 2-chloro-4-methylquinazoline (357 mg, 2.0 mmol) and M36 (526 mg, 2.0 mmol) as raw materials, and referring to the synthesis method of intermediate M33a, a pale 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] + calcd for C 22 H 20 N5O2F 406.1674, found 406.1675.

[0161] The synthesis of intermediate M37 was as follows: Compound C4 (520 mg, 1.28 mmol) was dissolved in methanol (20 mL) and water (5 mL) in a 125 mL round-bottom flask, and sodium hydroxide (154 mg, 3.84 mmol) was added. The reaction mixture was stirred overnight at 60 °C, and the reaction was monitored by TLC. After the reaction was complete, the crude product was distilled under reduced pressure to obtain an aqueous solution. The pH was adjusted to 3-4 with the addition of saturated citric acid solution, and a solid precipitated. The solid was filtered, the filter cake was washed with water, and dried in air to obtain a white solid intermediate compound M37 (460 mg, 95% yield). 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).

[0162] The synthesis of compound C5 was as follows: In a 50 mL flask, intermediate M37 (100 mg, 0.26 mmol) was dissolved in anhydrous N,N-dimethylformamide (8 mL). Compound methylamine hydrochloride (24 mg, 0.29 mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (121 mg, 0.32 mmol), and diisopropylethylamine (103 mg, 0.78 mmol) were added sequentially. The reaction mixture was stirred overnight at room temperature, and the reaction was monitored by TLC. After the reaction was complete, the crude product was obtained by vacuum distillation and purified by silica gel column chromatography to give a pale yellow solid C5 (81 mg, 76% yield). 1 H NMR (400MHz, CDCl3) δ8.43(dd,J=4.0,2.4Hz,1H),7.86(d,J=8.4Hz,1H),7.68–7.61(m,2H),7.58(dd,J=10.0,5.2Hz ,1H),7.25–7.14(m,2H),4.72–4.53(m,4H),4.22–4.06(m,1H),3.15(s,6H),2.79(s,3H).HRMS(ESI-TOF)m / z:[M+H] + calcd for C 22 H 21 N6OF 405.1834,found405.1846.

[0163] The synthesis of intermediates M38a-c was as follows: In a 50 mL round-bottom flask, intermediate M37 (100 mg, 0.26 mmol) was dissolved in anhydrous N,N-dimethylformamide (8 mL). Various amines (0.29 mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (121 mg, 0.32 mmol), and diisopropylethylamine (103 mg, 0.78 mmol) were added sequentially. The reaction mixture was stirred overnight at room temperature, and the reaction was monitored by TLC. After the reaction was complete, the crude product was obtained by vacuum distillation, and purified by silica gel column chromatography to give the yellow solid intermediate compounds M38a-c.

[0164] 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).

[0165] 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).

[0166] Intermediate M38c, 230mg, 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).

[0167] 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] + calcd for C 24 H 24 N7OF 446.2099, found 446.2098.

[0168] 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] + calcd for C 25 H 26 N7OF 446.2099,found446.2098.

[0169] 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] + calcd for C 26 H 26 N7OF 472.2256, found 472.2251.

[0170] In this embodiment, the synthetic route for compounds C9 to C10 (see Figure 19) is as follows:

[0171] The synthesis of compound C9 was as follows: 2-Chloroquinoxaline (164 mg, 1.0 mmol) and intermediate M3a (245 mg, 1.0 mmol) were dissolved in anhydrous DMF (10 mL) in a 125 mL flask, and cesium carbonate (977 mg, 3.0 mmol) was added. The reaction mixture was heated to 110 °C for 6 h, and the reaction was monitored by TLC. After the reaction was complete, the cesium carbonate was removed by filtration, and the filtrate was distilled under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography to give a pale yellow solid compound C9 (186 mg, 50% yield). 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).

[0172] The synthesis of compound C10 was carried out as follows: using 2-chloroquinoxaline (100 mg, 0.61 mmol) and intermediate M17e (159 mg, 0.61 mmol) as raw materials, and following the synthesis method of compound C9, a pale yellow solid compound C10 (128 mg, yield 54%) was obtained. 1H NMR (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).

[0173] The following tests were performed on the pyrazolopyridine compounds linked to the aza-four-membered ring in each embodiment:

[0174] Inhibition test of PDE10A (phosphodiesterase type 10A) activity by pyrazolopyridine compounds linked by aza-four-membered rings

[0175] At room temperature, the pyrazolopyridine compound to be tested, linked to a four-membered ring, was reacted with 20,000–30,000 cpm of a solution containing 1.0 μg / mL recombinant clone PDE10A protein, 20 mM Tris-HCl (pH 7.5), 4.0 mmol / L dithiothreitol, and 10 mmol / L MgCl2. 3 The H-cAMP mixture was 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 ions in the supernatant were then measured using a PerkinElmer 2910 counter. 3 H-cAMP was used, and the experiment was repeated at least three times. The IC50 value for the inhibition of recombinant clone PDE10A (phosphodiesterase type 10A) protein activity by aza-4-membered ring-linked pyrazolopyridine compounds was obtained through concentration testing and nonlinear regression calculations. 50 value.

[0176] The results of the activity inhibition tests of the aza-4-membered ring-linked pyrazolopyridine compounds against the recombinant clone PDE10A protein in each embodiment are shown in Table 1. Under the same conditions, the activity inhibition test results of the positive control (papaverine) against the recombinant clone PDE10A protein should be controlled within IC50. 50 =In the range of 50 to 100 nmol / L, to ensure the IC50 of the measured compound. 50 The value data has a unified reference standard.

[0177] Table 1. Inhibitory activity of PDE10A protein by pyrazolidine compounds linked by aza-four-membered rings.

[0178] Example 2: Confirmatory Experiment of PDE10A Inhibitor Treatment for Pulmonary Hypertension in a Wild Lilium-Induced PAH Animal Model

[0179] Experimental Procedure: The experiment used the PDE10A inhibitor sample A11 as an example. The experiment was divided into 6 groups (one group each of 1.25, 2.5, and 5.0 mg / kg A11, a positive control group (Tadalafil 5.0 mg / kg), a model group, and a normal control group), with 12 rats in each group. Except for the normal control group, which received saline, the rats in the other groups received a single intraperitoneal injection of 2% limonene alkaloid 60 mg / kg to establish a pulmonary hypertension model. On the second day, the following administration was initiated: (1) The normal control group and the model group were given blank solvent [1.6% DMSO + 98.4% CMC-Na (0.5%)] by gavage daily; (2) The A11 test drug group was given oral administration of 1.25 mg / kg (low dose group), 2.5 mg / kg (medium dose group), and 5.0 mg / kg (high dose group) daily; (3) All animals were continuously administered 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. During the experiment, the rats' body weight was measured every three days, and their general condition (respiration, activity, etc.) was observed. One hour after the last administration, the animals were anesthetized intraperitoneally with 3% sodium pentobarbital, and the pulmonary artery pressure of the rats was measured using the right heart catheterization method.

[0180] Measurement of rat pulmonary artery pressure (mPAP): After intraperitoneal anesthesia, the rats were fixed in a supine position. A longitudinal incision was made at the right clavicle of the neck, and the surrounding tissues were bluntly dissected. The right external jugular vein was carefully dissected using mosquito forceps, and the vein was freed for about 1 cm. Two sutures were passed below the vein; the distal suture was ligated to the vein, and the proximal suture was loosely knotted for later use. The proximal and distal sutures were fixed to the skin of the rat's chest and neck with hemostatic forceps to ensure full exposure and filling of the vein. The physiological recorder was adjusted and calibrated, while keeping the sensor parallel to the rat's heart. Make a V-shaped incision near the heart using ophthalmic scissors. Insert the curved end of the right heart catheter into the vascular incision. Secure the catheter in place with a slipknot, and slowly advance the catheter. Insertion of 1-2 cm will reach the superior vena cava, and 2-3 cm will reach the right atrium. Gently rotate and advance the catheter until it reaches the right ventricle, approximately 3.5-4 cm. The right ventricular systolic pressure (RVSP) can then be measured. Pause briefly, then gently push the catheter into the pulmonary artery. During insertion, observe the pressure curve waveform on the monitor to determine the catheter tip position and calculate the mean pulmonary artery pressure using the pulmonary artery systolic and diastolic pressure values.

[0181] Right ventricular hypertrophy index (RVHI): After the pulmonary hemodynamic parameters are measured, the skin of the neck and chest is cut open to expose the trachea. The thoracic cavity is quickly dissected to expose the heart and lungs. The trachea is clamped with hemostatic forceps, cut open, and the heart, thymus, and lungs are removed together. The heart is removed, and the blood vessels and atria are removed. The right ventricle (RV) is freed, leaving the left ventricle and interventricular septum (LV+S). The blood is washed away with saline and the water is absorbed with filter paper. The weight of the right ventricle (RV) and the left ventricle plus the interventricular septum (LV+S) is weighed, and the ratio of RV / (LV+S) is used as the right ventricular hypertrophy index.

[0182] Compared to the model group, the aza-4-membered ring-linked pyrazolopyridine PDE10A inhibitor A11 of this invention significantly reduced the mean pulmonary artery pressure and right ventricular systolic pressure in rats with limonene-induced pulmonary hypertension (see Figure 1), while simultaneously improving pulmonary arteriolar intimal hyperplasia (thickening) caused by pulmonary hypertension and significantly improving right ventricular hypertrophy caused by pulmonary hypertension (see Figure 2). Lung tissue section staining, including hematoxylin-eosin H&E, Masson's, and α-smooth muscle actin (α-SMA) (see Figure 3), further demonstrates that the aza-4-membered ring-linked pyrazolopyridine PDE10A inhibitor of this invention can improve the pathological remodeling of pulmonary arteriolar vessels and exert a therapeutic effect on pulmonary hypertension.

[0183] Example 3: Confirmatory Experiment on PDE10A Inhibitor Treatment of Pulmonary Hypertension in a PAH Animal Model under Hypoxic Conditions 1. Animal Experiment and Administration Route

[0184] Taking the PDE10A inhibitor sample A11 as an example, C57 mice (20g, 56 mice) were acclimatized for three days before the experiment. Afterwards, they were randomly divided into groups under normoxic (20% O2) and hypoxic (10% O2) conditions: normoxic control group, SU5416 (Sugen, synthetic, purity >99%) + hypoxia (model group), and SU5416 + hypoxia administration groups (tadalafil group, low, medium, and high dose A11 groups). On day 1, mice in the model group and each administration group received a single intraperitoneal injection of 20mg / kg SU5416 followed by hypoxic environment feeding (21 days). Supplementary injections of SU5416 (20mg / kg) were given on days 8 and 16. The normoxic control group was fed for 21 days. All mice were provided with 12 hours of light and dark environment, as well as food and water, and were weighed every three days. Specific dosages are as follows:

[0185] 2. Right ventricular systolic pressure (RVSP) of mice was measured by jugular vein cannulation on day 22. 1) Machine preparation: Select the tidal volume for mice on the small animal ventilator, then connect the computer to the BL-420N biosignal acquisition and processing system, select a single channel, purge bubbles from the tubing, and zero the device; 2) Anesthesia: Weigh the mouse, anesthetize it with an intraperitoneal injection of 1% sodium pentobarbital solution (20mg / kg), lay the mouse flat, check if 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's neck and chest cavity with medical alcohol; 4) Intubation: Make a 1cm straight incision at the base of the mouse's neck with a scalpel, separate the subcutaneous tissue to expose the trachea, bluntly and sharply separate the soft tissue around the trachea with ophthalmic scissors and forceps, free the trachea, make a "V" shaped incision about 2mm long with ophthalmic scissors, insert the trachea, and observe that the rise and fall of the mouse's chest cavity is consistent with the ventilator; 5) Pressure measurement: Cut open the chest skin with scissors to expose the subcutaneous tissue, and be careful to bluntly separate the tissue to avoid accidental injury. Then, lift the xiphoid process with forceps, cut open the chest cavity with scissors, and tear open the pericardium with forceps to expose the heart. Insert the tubing connected to the syringe into the right ventricle. Display the right ventricular pressure waveform on the computer for 10 seconds, record the data, and save it.

[0186] 3. Subsequent thoracotomy and perfusion of the mouse heart and lung tissue. After measuring right ventricular pressure, physiological saline containing heparin sodium was injected into the heart through the right ventricle to flush the heart and lung tissue, allowing blood to drain until the flushed fluid became clear and the lungs turned white. Pericardial adipose tissue was removed, and the superior and inferior vena cava, aorta, pulmonary artery, and pulmonary veins were severed. Left and right ventricular tissues were extracted and weighed. 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; the other two portions were cryopreserved separately. Two portions of the right ventricle were taken. One portion was fixed with paraformaldehyde and sent to the company for sectioning and hematoxylin-eosin H&E and wheat germ agglutinin (WGA) staining; the other portion was cryopreserved for later use.

[0187] 4. Results: Animal experiments showed that, compared with the model group, the high-dose group of the aza-4-membered ring-linked pyrazolopyridine PDE10A inhibitor A11 of this invention significantly alleviated the weight loss in rats after modeling (see Figure 4), reduced right ventricular systolic pressure RVSP (see Figure 5A) and right ventricular hypertrophy index (see Figure 5B), and improved pulmonary arteriolar intimal hyperplasia caused by pulmonary hypertension (see Figure 6A). The efficacy was similar to that of the positive control tadalafil at the same dose. WGA staining of the right ventricle showed that the high-dose A11 group significantly alleviated cardiomyocyte hypertrophy in rats after modeling, with an effect similar to tadalafil (see Figure 6B). HE staining of lung tissue showed that the high-dose A11 group significantly inhibited SU5416+ hypoxia-induced vascular wall thickening in rats (see Figure 7).

[0188] In summary, the pyrazolopyridine PDE10A inhibitor linked by the aza-four-membered ring of the present invention can be used as a drug molecule for the treatment of pulmonary hypertension.

[0189] The technical solutions of the present invention are not limited to the specific embodiments described above. Any technical modifications made in accordance with the technical solutions of the present invention fall within the protection scope of the present invention.

Claims

1. The application of pyrazolopyridine compounds linked by a nitrogen-containing four-membered ring in the preparation of drugs for treating pulmonary hypertension, characterized in that: The aza-quaternary ring-linked pyrazolopyridine compounds have the structure shown in formula (I): wherein: R1, R2, R3substituents are each independently H, halogen or 18 F, C 1-3 alkyl, C 1-3 alkoxy, difluoromethoxy, difluoroethoxy, trifluoromethyl, trifluoromethoxy, acetyl, cyano; Quinoxaline R6- or R7-substituted quinolines and R6, R 10 Substituted quinazoline One of them; R4 is One of them; R5 represents H and C. 1-5 Alkyl, isopropyl, -C (1-3) CH2OH, -C (1-3) One of CH2OCH3, -C(=O)CH3, -SO2CH3, and -C(=O)OCH3; R6 is an F atom that is either monosubstituted or disubstituted. 18 One of the following: F atom, Cl atom, hydroxyl group, methyl group, trifluoromethyl group, methoxy group, and cyano group; R7 can be independently hydroxyl, methyl, trifluoromethyl, or C. 1-3 One of the following: alkoxy group, R4, -C(=O)-R4, R8, -CH2-R9, -C(=O)CH2-R9, -NHC(=O)-CH2-R9; R8 is: One of them; R9 is One of them; R 10 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, The compound comprising a pyrazolopyridine compound linked by a nitrogen-containing four-membered ring as described in claim 1.

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