A class of fused heterocyclic compounds, methods of making and uses thereof

Abstract

The application discloses a kind of fused heterocyclic compounds, preparation method and purposes thereof.The specific, the application discloses a kind of compound shown in formula (I) for treating cardiovascular disease, its isotope marker, enantiomer, diastereoisomer, atropisomer or its pharmaceutically acceptable salt.The compound of the application can be used as NPR1 positive allosteric modulator, and has good application prospect.

Description

[0001] This application claims priority to Chinese Patent Application No. 2025101141626, filed January 24, 2025, and Chinese Patent Application No. 2025106282724, filed May 15, 2025. The full text of the aforementioned Chinese patent applications is incorporated herein by reference. Technical Field

[0002] This invention relates to an NPR1 modulator, its preparation method and application, and more specifically to a class of fused heterocyclic compounds, their compositions and their use in the treatment and prevention of cardiovascular diseases. Background Technology

[0003] Cardiovascular disease is the leading cause of death worldwide today, with approximately 20.5 million people dying from it in 2021, accounting for one-third of all deaths globally (Lindstrom M, et al.). J Am Coll Cardiol, 2022, 80(25): 2372-2425). As a common cardiovascular disease, hypertension, which causes elevated blood pressure and reduced blood flow, can lead to heart disease, heart failure, stroke, kidney damage, and kidney failure. According to the World Health Organization, there are 1.28 billion people aged 30-79 with hypertension worldwide.

[0004] Natriuretic peptide receptor type A (NPR1) is a type of guanylate cyclase receptor. When the ligands atrial natriuretic peptide (ANP) or brain natriuretic peptide (BNP) bind to the extracellular receptor domain of NPR1, a conformational change in the protein is induced, transmitting an activation signal to the intracellular guanylate cyclase domain, thereby catalyzing the production of 3',5'-cyclic guanosine monophosphate as a second messenger to regulate blood pressure. In heart failure, natriuretic peptides released by increased intracardiac pressure and cardiomyocyte stretching act by binding to NPR1, affecting vasodilation, natriuresis, diuresis, reducing venous pressure, and relieving venous congestion. NPR1 knockout mice have been reported to have hypertension, and ANP-deficient mice have also been reported to have salt-sensitive hypertension. In addition, endothelial NPR1-specific knockout mice have been found to have cardiac hypertrophy and cardiac fibrosis in addition to hypertension (John SWM, et al.). Science, 1995, 267(5198):679-681; Sackner-Bernstein JD, et al. JAMA, 2005, 293(15): 1900-1905). Therefore, NPR1 plays an important physiological role in regulating blood pressure and maintaining heart health. Currently, recombinant ANP (Carperitide) and BNP (Nesiritide) products have been approved for the treatment of acute heart failure. Studies have found that patients treated with nesiritide have better prognoses than those treated with cardiotonics (Mitaka C, et al. Crit Care, 2011, 15(5): 1-10). However, due to the short half-life and poor oral bioavailability of peptide drugs, they are difficult to apply to the control and treatment of hypertension. Therefore, developing oral small molecule drugs targeting NPR1 can meet the treatment needs of more patients. Summary of the Invention

[0005] To overcome the shortcomings of existing technologies, this invention provides a class of compounds, their preparation methods, and applications. The compounds of this invention can serve as positive allosteric modulators of NPR1.

[0006] This invention provides a compound of formula (I), its isotopic label, enantiomer, diastereomer, transisomer, or pharmaceutically acceptable salt thereof:

[0007] in, It can be a single bond or a double bond; X 1 For N, NR x1 CR 1 or CR 1 R 1a X 2 For N, NR x2 CR 2 or CR 2 R 2a X 3 For N or CR 3 X 4 For N or CR 4 X 5 For N or CR 5 X 6 For N or CR 6 ; R x1 and R x2 Each is independently selected from hydrogen, C1-C6 alkyl, halo-C1-C6 alkyl, and deuterated C1-C6 alkyl; R1 R 1a R 2 R 2a R 3 R 4 R 5 R 6 Each is independently selected from hydrogen, deuterium, halogen, cyano, carboxyl, hydroxyl, -NR x R y -C(=O)NR x R y -NR x C(=O)R y -C(=O)OR z -C(=O)R z -S(=O)2R z -C(=O)NH-S(=O)2R z -P(=O)R z R t No substitution or by one or more R A Substituted C1-C6 alkyl, unsubstituted, or with one or more R A Substituted C1-C6 alkoxy groups, unsubstituted, or substituted with one or more R groups A Substituted C2-C6 alkenyl groups, unsubstituted, or with one or more R groups A Substituted C2-C6 ynyl group, unsubstituted, or with one or more R groups A Substituted C3-C6 cycloalkyl, unsubstituted, or with one or more R A Substituted C6-C8 aryl groups, unsubstituted or with one or more R groups A Substituted 4-6 membered heterocyclic groups, unsubstituted or with one or more R groups A Substituted 5-6 aryl groups; R A Selected from C1-C6 alkyl, halogenated C1-C6 alkyl, deuterated C1-C6 alkyl, C1-C6 alkoxy, halogenated C1-C6 alkoxy, deuterated C1-C6 alkoxy, deuterium, halogen, carboxyl, hydroxyl, cyano, -NR A1 R A2 -C(=O)NR A1 R A2 -C(=O)OR A1 -NR A1 C(=O)R A2 ; L 1 Selected from , or ;in, Indicates with L 2 The connected side; R L1 It is hydrogen or C1-C6 alkyl; L 2 -(CR) 7 R 8 ) i -; Each R 7 and R 8 Each of these can be independently hydrogen, deuterium, halogen, cyano, carboxyl, hydroxyl, C1-C6 alkyl, halo-C1-C6 alkyl, or deuterated C1-C6 alkyl; i can be 0, 1, 2, or 3. W is selected from unsubstituted or R-selected. B Substituted C6-C10 cycloalkyl, unsubstituted, or with one or more R B Substituted 5-10 membered heterocyclic groups; the 5-10 membered heterocyclic group contains 1-2 N atoms; R B Selected from oxo (=O), deuterium, halogen, hydroxyl, cyano, -NR B1 R B2 -C(=O)NR B1 R B2 -C(=O)OR B1 -NR B1 C(=O)R B2 、-(CR 9 R 10 ) n -R 11 -OR 12 ; n is 0, 1, 2, or 3; each R 9 and R 10 Each can be independently hydrogen, deuterium, halogen, cyano, carboxyl, hydroxyl, C1-C6 alkyl, halo-C1-C6 alkyl, or deuterated C1-C6 alkyl. R 11 and R 12 Selected from unsubstituted or by one or more R C Substituted C1-C6 alkyl, unsubstituted, or with one or more R C Substituted C2-C6 alkenyl groups, unsubstituted, or with one or more R groups C Substituted C2-C6 ynyl group, unsubstituted, or with one or more R groups C Substituted C3-C6 cycloalkyl, unsubstituted, or with one or more R C Substituted C6-C10 aryl groups, unsubstituted, or substituted with one or more R groups C Substituted 4-6 membered heterocyclic groups, unsubstituted or with one or more R groups C Substituted 5-6 aryl groups; R CSelected from oxo (=O), deuterium, halogen, hydroxyl, cyano, C1-C6 alkyl, deuterated C1-C6 alkyl, halo-C1-C6 alkyl, hydroxyl-substituted C1-C6 alkyl, cyano-substituted C1-C6 alkyl, C1-C6 alkoxy-substituted C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, halo-C1-C6 alkoxy, C3-C6 cycloalkyl, C3-C6 cycloalkoxy, -NR C1 R C2 -C(=O)NR C1 R C2 -C(=O)OR C1 -NR C1 C(=O)R C2 -SO2R C1 ; Or two adjacent R B Together with the atoms between them, they form unsubstituted or substituted groups with one or more R atoms. D Substituted phenyl, unsubstituted, or substituted with one or more R D Substituted 5-6 aryl groups; R D Selected from C1-C6 alkyl, halogenated C1-C6 alkyl, deuterated C1-C6 alkyl, C1-C6 alkoxy, halogenated C1-C6 alkoxy, deuterated C1-C6 alkoxy, deuterium, halogen, hydroxyl, carboxyl, cyano, -NR D1 R D2 -C(=O)NR D1 R D2 -NR D1 C(=O)R D2 ; R x R y R A1 R A2 R B1 R B2 R C1 R C2 R D1 R D2 Each is independently selected from hydrogen, C3-C6 cycloalkyl, unsubstituted, or by one or more R E Substituted C1-C6 alkyl groups; R E Selected from deuterium, halogens, C1-C6 alkoxy groups, deuterated C1-C6 alkoxy groups, halogenated C1-C6 alkoxy groups, and -NR. E1 R E2 ;R E1 R E2 Each is independently selected from hydrogen, C1-C6 alkyl, and C3-C6 cycloalkyl; R z R tEach is independently selected from C1-C6 alkyl groups; The 4-6 membered heterocyclic group contains 1-3 heteroatoms selected from N, O, and S, and the 5-6 membered heteroaryl group contains 1-3 heteroatoms selected from N, O, and S.

[0008] As a preferred technical solution, the compound represented by formula (I) has the following structure:

[0009] Among them, X 2 X 4 R 1 R 3 R 5 R 6 L 1 L 2 The definitions of W and W are the same as described above.

[0010] As a preferred technical solution, the compound represented by formula (I) has the following structure:

[0011] Among them, R 1 R 2 R 3 R 4 R 5 R 6 L 1 L 2 The definitions of W and W are the same as described above.

[0012] As a preferred technical solution, R 1 R 2 R 3 R 4 R 5 R 6 Each is independently selected from hydrogen, deuterium, halogen, cyano, carboxyl, hydroxyl, amino, C1-C6 alkyl, C1-C6 alkoxy, halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, deuterated C1-C6 alkyl, and deuterated C1-C6 alkoxy.

[0013] As a preferred technical solution, Selected from: , , , , , , , , .

[0014] As a preferred technical solution, L 1 for or L 2 For a bond or -CH2-; where, Indicates with L 2 The connected side.

[0015] As a preferred technical solution, L 1 for L 2 As the key; where, Indicates with L 2 The connected side.

[0016] As a preferred technical solution, W is selected from unsubstituted or R-1 or R-2. B The following groups are substituted: , , , , , , , , , , , , , , , , , , , ; R B Selected from deuterium, halogen, hydroxyl, cyano, -NR B1 R B2 -C(=O)NR B1 R B2 -C(=O)OR B1 -NR B1 C(=O)R B2 、-(CR 9 R 10 ) n -R 11 -OR 12 ; n is 0, 1, 2, or 3; each R 9 and R 10 Each can be independently hydrogen, deuterium, halogen, cyano, carboxyl, hydroxyl, C1-C6 alkyl, halo-C1-C6 alkyl, or deuterated C1-C6 alkyl. R11 and R 12 Selected from unsubstituted or by one or more R C Substituted C1-C6 alkyl, unsubstituted, or with one or more R C Substituted C2-C6 alkenyl groups, unsubstituted, or with one or more R groups C Substituted C2-C6 ynyl group, unsubstituted, or with one or more R groups C Substituted C3-C6 cycloalkyl, unsubstituted, or with one or more R C Substituted C6-C10 aryl groups, unsubstituted, or substituted with one or more R groups C Substituted 4-6 membered heterocyclic groups, unsubstituted or with one or more R groups C Substituted 5-6 aryl groups; R C Selected from oxo (=O), deuterium, halogen, hydroxyl, cyano, C1-C6 alkyl, deuterated C1-C6 alkyl, halo-C1-C6 alkyl, hydroxyl-substituted C1-C6 alkyl, cyano-substituted C1-C6 alkyl, C1-C6 alkoxy-substituted C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, halo-C1-C6 alkoxy, C3-C6 cycloalkyl, C3-C6 cycloalkoxy, -NR C1 R C2 -C(=O)NR C1 R C2 -C(=O)OR C1 -NR C1 C(=O)R C2 -SO2R C1 ; R B1 R B2 R C1 R C2 Each is independently selected from hydrogen, C3-C6 cycloalkyl, unsubstituted or modified by one or more R E Substituted C1-C6 alkyl groups; R E Selected from deuterium, halogens, C1-C6 alkoxy groups, deuterated C1-C6 alkoxy groups, halogenated C1-C6 alkoxy groups, and -NR. E1 R E2 ;R E1 R E2 Each is independently selected from hydrogen, C1-C6 alkyl, and C3-C6 cycloalkyl; The 4-6 membered heterocyclic group contains 1-3 heteroatoms selected from N, O, and S, and the 5-6 membered heteroaryl group contains 1-3 heteroatoms selected from N, O, and S.

[0017] As a preferred technical solution, W is selected from: , , , , , , , , , , , , , , , , , , , ; m can be 0, 1, 2, 3, or 4; R 13 Selected from deuterium, halogen, hydroxyl, cyano, -NR 13A R 13B -C(=O)NR 13A R 13B -C(=O)OR 13A -NR 13A C(=O)R 13B 、-(CR 15 R 16 ) p -R 17 -OR 18 ; p is 0, 1, 2, or 3; each R 15 and R 16 Each can be independently hydrogen, deuterium, halogen, cyano, carboxyl, hydroxyl, C1-C6 alkyl, halo-C1-C6 alkyl, or deuterated C1-C6 alkyl. R 17 and R 18 Selected from unsubstituted or by one or more R 19 Substituted C1-C6 alkyl, unsubstituted, or with one or more R 19 Substituted C2-C6 alkenyl groups, unsubstituted, or with one or more R groups 19 Substituted C2-C6 ynyl group, unsubstituted, or with one or more R groups 19 Substituted C3-C6 cycloalkyl, unsubstituted, or with one or more R 19 Substituted C6-C10 aryl groups, unsubstituted, or substituted with one or more R groups 19 Substituted 4-6 membered heterocyclic groups, unsubstituted or with one or more R groups 19 Substituted 5-6 aryl groups; R 19Selected from oxo (=O), deuterium, halogen, hydroxyl, cyano, C1-C6 alkyl, deuterated C1-C6 alkyl, halo-C1-C6 alkyl, hydroxyl-substituted C1-C6 alkyl, cyano-substituted C1-C6 alkyl, C1-C6 alkoxy-substituted C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, halo-C1-C6 alkoxy, C3-C6 cycloalkyl, C3-C6 cycloalkoxy, -NR 19A R 19B -C(=O)NR 19A R 19B -C(=O)OR 19A -NR 19A C(=O)R 19B ; R 14 For hydrogen or -(CR) 20 R 21 ) q -R 22 ; q is 0, 1, 2, or 3; each R 20 and R 21 Each can be independently hydrogen, deuterium, halogen, cyano, carboxyl, hydroxyl, C1-C6 alkyl, halo-C1-C6 alkyl, or deuterated C1-C6 alkyl. R 22 Selected from unsubstituted or by one or more R 23 Substituted C1-C6 alkyl, unsubstituted, or with one or more R 23 Substituted C3-C6 cycloalkyl, unsubstituted, or with one or more R 23 Substituted 4-6 membered heterocyclic groups, unsubstituted or with one or more R groups 23 Substituted 5-6 aryl groups; R 23 Selected from oxo (=O), deuterium, halogen, hydroxyl, cyano, C1-C6 alkyl, deuterated C1-C6 alkyl, halo-C1-C6 alkyl, hydroxyl-substituted C1-C6 alkyl, cyano-substituted C1-C6 alkyl, C1-C6 alkoxy-substituted C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, halo-C1-C6 alkoxy, C3-C6 cycloalkyl, C3-C6 cycloalkoxy, -NR 23A R 23B -C(=O)NR 23A R 23B -C(=O)OR 23A -NR 23A C(=O)R 23B -SO2R 23A ; R 13A R 13B R 19AR 19B R 23A R 23B Each is independently selected from hydrogen, C3-C6 cycloalkyl, unsubstituted, or by one or more R F Substituted C1-C6 alkyl groups; R F Selected from deuterium, halogens, C1-C6 alkoxy groups, deuterated C1-C6 alkoxy groups, halogenated C1-C6 alkoxy groups, and -NR. F1 R F2 ;R F1 R F2 Each is independently selected from hydrogen, C1-C6 alkyl, and C3-C6 cycloalkyl; The 4-6 membered heterocyclic group contains 1-3 heteroatoms selected from N, O, and S, and the 5-6 membered heteroaryl group contains 1-3 heteroatoms selected from N, O, and S.

[0018] As a preferred technical solution, W is selected from: , , , , , , , , , , , , , , , , , , , ; m can be 0, 1, 2, 3, or 4; R 13 Selected from halogen, hydroxyl, cyano, -NR 13A R 13B -C(=O)NR 13A R 13B -C(=O)OR 13A -NR 13A C(=O)R 13B 、-(CR 15 R 16 ) p -R 17 -OR 18 ; p is 0, 1, 2, or 3; each R 15 and R 16 Each is independently hydrogen or C1-C6 alkyl; R 17and R 18 Selected from unsubstituted or by one or more R 19 Substituted C1-C6 alkyl, unsubstituted, or substituted with one or more R 19 Substituted C2-C6 alkenyl groups, unsubstituted, or with one or more R groups 19 Substituted C2-C6 ynyl group, unsubstituted, or with one or more R groups 19 Substituted C3-C6 cycloalkyl, unsubstituted, or with one or more R 19 Substituted C6-C10 aryl groups, unsubstituted, or substituted with one or more R groups 19 Substituted 4-6 membered heterocyclic groups, unsubstituted or with one or more R groups 19 Substituted 5-6 aryl groups; R 19 Selected from oxo (=O), deuterium, halogen, hydroxyl, cyano, C1-C6 alkyl, deuterated C1-C6 alkyl, halo-C1-C6 alkyl, hydroxyl-substituted C1-C6 alkyl, cyano-substituted C1-C6 alkyl, C1-C6 alkoxy-substituted C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, halo-C1-C6 alkoxy, C3-C6 cycloalkyl, C3-C6 cycloalkoxy, -NR 19A R 19B -C(=O)NR 19A R 19B -C(=O)OR 19A -NR 19A C(=O)R 19B ; R 13A R 13B R 19A R 19B Each is independently selected from hydrogen, C3-C6 cycloalkyl, unsubstituted, or by one or more R... F Substituted C1-C6 alkyl groups; R F Selected from deuterium, halogens, C1-C6 alkoxy groups, deuterated C1-C6 alkoxy groups, halogenated C1-C6 alkoxy groups, and -NR. F1 R F2 ;R F1 R F2 Each is independently selected from hydrogen, C1-C6 alkyl, and C3-C6 cycloalkyl; The 4-6 membered heterocyclic group contains 1-3 heteroatoms selected from N, O, and S, and the 5-6 membered heteroaryl group contains 1-3 heteroatoms selected from N, O, and S; R 14 Selected from hydrogen, -CH2N(CH3)2, -CH2CH2N(CH3)2, -CH2CH2CH2N(CH3)2, , , , , , , , , , , , , , , , , -SO2CH3.

[0019] As a preferred technical solution, W is selected from: , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , .

[0020] As a preferred technical solution, the compound shown in formula (I) is selected from the following structures:

[0021] Wherein, s is 0, 1 or 2;<000> t is 0, 1 or 2; X 2 is N or CR 2 , X 4 is N or CR 4 ; R 1 , R 2 , R 3 , R 4 , R 5 , R 6 are each independently selected from hydrogen, deuterium, halogen, cyano, carboxyl, hydroxyl, amino, C1-C6 alkyl, C1-C6 alkoxy, halo C1-C6 alkyl, halo C1-C6 alkoxy, deuterated C1-C6 alkyl, deuterated C1-C6 alkoxy; R 24 is hydrogen or -(CR 25 R 26 ) y -R 27 ; y is 0, 1, 2 or 3; each R 25 and R 26 are each independently hydrogen, deuterium, halogen, cyano, carboxyl, hydroxyl, C1-C6 alkyl, halo C1-C6 alkyl, deuterated C1-C6 alkyl; R 27 is selected from unsubstituted or substituted by one or more R 28 substituted C1-C6 alkyl, unsubstituted or substituted by one or more R 28 [[ID=(69)]]substituted 4-6 membered heterocyclic group, unsubstituted or substituted by one or more R 28 substituted 5-6 membered heteroaryl; R 28 is selected from oxo (=O), deuterium, halogen, hydroxyl, cyano, C1-C6 alkyl, deuterated C1-C6 alkyl, halo C1-C6 alkyl, hydroxyl-substituted C1-C6 alkyl, cyano-substituted C1-C6 alkyl, C1-C6 alkoxy-substituted C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, halo C1-C6 alkoxy, C3-C6 cycloalkyl, C3-C6 cycloalkoxy, -NR 28A R 28B, -C(=O)NR 28A R 28B , -C(=O)OR 28A , -NR 28A C(=O)R 28B , -SO2R 28A ;[[ID=1)3]] R 28A 、R 28B are each independently selected from hydrogen, C3-C6 cycloalkyl, unsubstituted or substituted by one or more R G substituted C1-C6 alkyl; R G is selected from deuterium, halogen, C1-C6 alkoxy, deuterated C1-C6 alkoxy, halogenated C1-C6 alkoxy, -NR G1 R G2 ; R G1 、R G2 are each independently selected from hydrogen, C1-C6 alkyl, C3-C6 cycloalkyl; The 4- to 6-membered heterocyclic group contains 1 to 3 heteroatoms selected from N, O, S, and the 5- to 6-membered heteroaryl group contains 1 to 3 heteroatoms selected from N, O, S.

[0022] As a preferred technical solution, R 24 is selected from hydrogen, -CH2N(CH3)2, -CH2CH2N(CH3)2, -CH2CH2CH2N(CH3)2, , , , , , , , , , , , , , , , , -SO2CH3.

[0023] As a preferred technical solution, the compounds represented by formula (I) are selected from the following compounds: 。

[0024] The present invention also provides a method for preparing a compound represented by formula (I), its isotope-labeled compound, enantiomer, diastereoisomer, atropisomer or its pharmaceutically acceptable salt.

[0025] When L 1 is the preparation method of the compound represented by formula (I) is Method 1; Method 1 includes the following steps: in the presence of a condensing agent (such as HATU, T3P, T4P or EDCI), reacting the following compound 1 and compound 2 in a solvent to obtain the compound represented by formula (I); ; wherein, X 1 , X 2 , X 3 , X 4 , X 5 , X 6 , R L1 , L2 and W are as defined above; When L 1 is the preparation method of the compound represented by formula (I) is Method 2; Method 2 includes the following steps: in the presence of a condensing agent (such as HATU, T3P, T4P or EDCI), reacting the following compound 1 and compound 2 in a solvent to obtain the compound represented by formula (I);

[0026] wherein, X 1 , X2 , X 3 , X 4 , X 5 , X 6 , R L1 , the definitions of L2 and W are the same as described above; When L 1 is , the preparation method of the compound shown in formula (I) is Method 3; Method 3 includes the following steps: in the presence of a catalyst (such as Pd2(dba)3, Pd(OAc)2, etc.) and a ligand (such as Xantphos, XPhos, Brettphos, etc.) and a base (such as Cs2CO3, K2CO3, t -BuOK, t -BuONa, etc.), reacting compound 1 and compound 2 in a solvent to obtain compound 3; or in the presence of a base (such as Cs2CO3, K2CO3, t -BuOK, t -BuONa, etc.), reacting compound 1 and compound 2 in a solvent to obtain compound 3. Compound 3 is alkylated or reductively aminated to obtain the compound shown in formula (I);

[0027] wherein, X 1 , X 2 , X 3 , X 4 , X 5 , X 6 , R L1 , the definitions of L2 and W are the same as described above.

[0028] The present invention also provides a pharmaceutical composition, which includes the compound shown in formula (I) as described above, its isotope-labeled compound, enantiomer, diastereomer, atropisomer or its pharmaceutically acceptable salt, and pharmaceutical excipients.

[0029] In the described composition, the dosage of the compound shown in formula (I) as described above, its isotope-labeled compound, enantiomer, diastereomer, atropisomer or its pharmaceutically acceptable salt can be an effective therapeutic amount.

[0030] The present invention also provides an application of the compound shown in formula (I) as described above, its isotope-labeled compound, enantiomer, diastereomer, atropisomer or its pharmaceutically acceptable salt in the preparation of an NPR1 regulator or a drug; the drug is a drug for treating and / or preventing diseases by regulating NPR1.

[0031] The present invention also provides the use of the compound as shown in formula (I) above, its isotope-labeled compound, enantiomer, diastereoisomer, atropisomer or pharmaceutically acceptable salt thereof in the preparation of a drug for treating and / or preventing cardiovascular diseases.

[0032] The present invention also provides a treatment method for treating and / or preventing cardiovascular diseases, which comprises administering to a patient a therapeutically effective amount of the compound as shown in formula (I) above, its isotope-labeled compound, enantiomer, diastereoisomer, atropisomer or pharmaceutically acceptable salt thereof.

[0033] The term "pharmaceutically acceptable" means relatively non-toxic, safe and suitable for use by patients.

[0034] The term "pharmaceutically acceptable salt" refers to a salt obtained by reacting a compound with a pharmaceutically acceptable acid or base. When a compound contains a relatively acidic functional group, the base addition salt can be obtained by contacting the compound with a sufficient amount of a pharmaceutically acceptable base in a suitable inert solvent. When a compound contains a relatively basic functional group, the acid addition salt can be obtained by contacting the compound with a sufficient amount of a pharmaceutically acceptable acid in a suitable inert solvent. For details, see Handbook of Pharmaceutical Salts: Properties, Selection, and Use (P. Heinrich Stahl, Camille G. Wermuth, 2011, 2nd Revised Edition).

[0035] The "-" at the end of a group means that the group is connected to the rest of the molecule through this site. For example, CH3-C(=O)- means acetyl group.

[0036] In " ", the wavy line indicates the connection point of the group to the rest of the molecule.

[0037] The term "halogen" refers to fluorine, chlorine, bromine or iodine.

[0038] The term "oxo group" means =O, where an oxygen atom replaces two hydrogens on the same carbon atom, that is, a carbonyl group replaces a methylene group.

[0039] The term "alkyl" refers to a straight-chain or branched-chain, saturated monovalent hydrocarbon group having a specified number of carbon atoms (for example, C2-C6). Alkyl includes but is not limited to: methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, n-hexyl, etc.

[0040] The term "alkoxy" refers to the group R X -O-, R XThe definition of is the same as the term "alkyl". Alkoxy groups include, but are not limited to: methoxy, ethoxy, n-propoxy, isopropoxy, etc.

[0041] The term "alkenyl" refers to a straight-chain or branched-chain, unsaturated monovalent hydrocarbon group having a specified number of carbon atoms (e.g., C1-C6), which has one or more (e.g., 1, 2 or 3) carbon-carbon sp 2 double bonds. Alkenyl groups include, but are not limited to: vinyl, , , etc.

[0042] The term "alkynyl" refers to a straight-chain or branched-chain, unsaturated monovalent hydrocarbon group having a specified number of carbon atoms (e.g., C2-C6), which has one or more (e.g., 1, 2 or 3) carbon-carbon triple bonds. Alkenyl groups include, but are not limited to: ethynyl, , , , etc.

[0043] The term "cycloalkyl" refers to a cyclic, saturated monovalent hydrocarbon group having a specified number of carbon atoms (e.g., C3-C⑥), which is a monocyclic, bridged-ring or spiro ring. Cycloalkyl groups include, but are not limited to: , , , , , , etc.

[0044] The term "cycloalkenyl" refers to an alicyclic hydrocarbon group having a specified number of carbon atoms (e.g., C3-C12, C3-C8, C3-C6) and containing 1, 2 or 3 double bonds. Cycloalkyl groups include, but are not limited to: [[ID=ID=38]], , etc.

[0045] The term "cycloalkoxy" refers to the group R Y -O-, where the definition of R Y is the same as the term "cycloalkyl". Cycloalkoxy groups include, but are not limited to, cyclopropoxy, etc.

[0046] The term "aryl" refers to a cyclic, unsaturated monovalent hydrocarbon group having a specified number of carbon atoms (e.g., C6-C 10 ), which is monocyclic or polycyclic (e.g., 2 or 3). When it is polycyclic, the monocyclic rings share two atoms and one bond, and each ring has aromaticity. Aryl groups include, but are not limited to: phenyl, naphthyl, etc.

[0047] In the present invention, the term "heterocyclic group" refers to a non-aromatic cyclic group containing at least one carbon atom and at least one (such as 1-3) ring heteroatom selected from N, O, and S, where the sulfur atom may be optionally oxidized or aminated. Examples of the "heterocyclic group" can specifically be groups formed by replacing one or more ring carbons of the cycloalkyl group as defined in the present invention with moieties selected from -O-, -N=, -NR-, -S-, -S(=O)-, and -S(=O)2-, where R is hydrogen, C 1-4 alkyl, C 3-6 cycloalkyl, or a nitrogen protecting group (for example, benzyloxycarbonyl, p-methoxybenzylcarbonyl, tert-butoxycarbonyl, acetyl, benzoyl, benzyl, p-methoxy-benzyl, p-methoxyphenyl, 3,4-dimethoxybenzyl, etc.). The "heterocyclic group" includes monocyclic and fused rings, bridged rings, spiro rings and other bicyclic structures and can be partially or completely saturated, such as 4-10 membered saturated or unsaturated heterocyclic groups, 4-6 membered saturated or unsaturated heterocyclic groups, 3-8 membered heterocyclic groups, 3-6 membered heterocyclic groups, etc.; such as tetrahydrofuranyl, pyrrolidinyl, oxetanyl, oxanyl, azetidinyl, oxiranyl, aziridinyl, thietanyl, 1,2-dithietanyl, 1,3-dithietyl, azepanyl, oxepanyl, etc. For example, the heterocyclic group described in the present invention can preferably be selected from the following groups: , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , or .

[0048] In the present invention, the term "heteroaryl" refers to a monocyclic, bicyclic or fused polycyclic ring aromatic hydrocarbon group having a specified number of ring atoms (e.g., 5-10 membered), which contains at least one (e.g., 1-3) ring heteroatom independently selected from N, O and S (e.g., N) in the ring, and the remaining ring atoms are carbon atoms; such as imidazolyl, pyridyl, pyrrolyl, thiazolyl, furyl, oxazolyl, isoxazolyl, pyrazolyl, thienyl, pyrimidinyl, 1,2,4-triazolyl, benzoxazolyl, imidazopyridyl, triazolopyridyl, benzofuryl, pyrazolopyrimidinyl, benzodioxolyl, indolyl, quinolinyl, isoquinolinyl, etc.

[0049] The term "heteroaromatic ring" satisfies at least one of the following conditions, and the remaining definitions are the same as those of the term "heteroaryl": 1. It is connected to the rest of the molecule by more than two single bonds; 2. It shares two atoms and one bond with the rest of the molecule.

[0050] The term "isotope label" refers to an isotope-labeled compound, in which one or more atoms in the isotope-labeled compound are replaced by atoms having an atomic mass or mass number different from the atomic mass or mass number of the atoms usually found in nature, compared with a compound represented by formula (I). Examples of isotopes that can be incorporated into the compounds of the present invention include isotopes of H, C, N, O, S, F and Cl, such as 2 H, 3 H, 13 C, 11 C, 14 C, 15 N, 18 O, 17 O, 32 P, 35 S, 18 F and 36Compounds of the present invention containing the above isotopes and / or other isotopes of other atoms, and pharmaceutically acceptable salts thereof. Some isotopically labeled compounds of the present invention, such as those incorporated with radioactive isotopes (such as 3 H and 14 C), can be used for drug and / or substrate tissue distribution assays. Tritium (i.e., 3 H) and carbon-14 (i.e., 14 C) isotopes are particularly preferred due to their ease of preparation and detectability. Furthermore, substitution with heavier isotopes (such as deuterium, i.e., 2 H or D) can provide certain therapeutic advantages (such as increased in vivo half-life or reduced dose requirements) stemming from higher metabolic stability, and can thus be preferred in certain cases. The compounds of the present invention can be specifically defined with substitution of deuterium or tritium. In addition, the omission of the separate listing of the terms deuterium or tritium for the hydrogen present in the substituents does not mean the exclusion of deuterium or tritium, but can also include deuterium or tritium.

[0051] The term "therapeutically effective amount" refers to the amount administered to a patient that is sufficient to effectively treat a disease. The therapeutically effective amount will vary depending on the type of compound, the type of disease, the severity of the disease, the age of the patient, etc., but can be adjusted by those skilled in the art according to the circumstances.

[0052] The term "pharmaceutical excipients" refers to all substances contained in a pharmaceutical preparation other than the active pharmaceutical ingredient, and is generally divided into two major categories: excipients and additives. For details, reference can be made to the Pharmacopoeia of the People's Republic of China (2020 Edition), Handbook of Pharmaceutical Excipients (Paul J Sheskey, Bruno C Hancock, Gary P Moss, David J Goldfarb, 2020, 9th Edition).

[0053] The term "treatment" refers to eliminating the cause or alleviating the symptoms.

[0054] The term "prevention" refers to reducing the risk of developing a disease.

[0055] The term "patient" refers to any animal that needs to receive treatment or prevention of a disease, usually a mammal, such as a human. Mammals include but are not limited to: cattle, horses, sheep, pigs, cats, dogs, mice, rats, rabbits, guinea pigs, monkeys, humans, etc.

[0056] On the basis of conforming to the common knowledge in the art, the above preferred conditions can be combined arbitrarily, and thus the preferred examples of the present invention can be obtained.

[0057] The reagents and raw materials used in the present invention are all commercially available.

[0058] The positive and progressive effects of the present invention are as follows: The compounds of the present invention can be used as NPR1 positive allosteric modulators for the treatment or prevention of cardiovascular diseases. Detailed implementation manners

[0059] The present invention will be further illustrated below by way of examples, but the present invention is not limited to the scope of the examples described herein. For the experimental methods without specific conditions noted in the following examples, they are carried out according to conventional methods and conditions, or selected according to the product specifications.

[0060] In each of the examples, 1 1H NMR was recorded on a BRUKER AVANCE NEO 400 MHz, JNM-ECZ400s nuclear magnetic resonance spectrometer, and the chemical shift was expressed in δ (ppm); liquid chromatography-mass spectrometry (LCMS) was recorded on a Shimadzu LC-20AD, Agilent 1260 mass spectrometer; preparative HPLC separation was performed using a WATERS 2545, Shimadzu LP-20AP liquid chromatograph.

[0061] In the following experimental descriptions, the following abbreviations may be used:

[0062] Example 1 Preparation of Compound I-2

[0063] Step 1: N -(2,5-Difluorophenyl)cinnamamide

[0064] 2,5-Difluoroaniline (15.0 g, 0.12 mol) and pyridine (18.4 g, 0.23 mol) were dissolved in dichloromethane (150 mL), and cinnamoyl chloride (20.3 g, 0.122 mol) was added dropwise to the solution at room temperature. The reaction solution was stirred at room temperature for 8 hours. The reaction solution was washed with saturated ammonium chloride (50 mL), the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and purified by silica gel chromatography (ethyl acetate / petroleum ether = 5%) to obtain N -(2,5-Difluorophenyl)cinnamamide (8.00 g, yield 26.6%). LCMS calc. for C 15 H 12 F2NO [M+H] + : m / z = 260.1; Found: 260.1.

[0065] Step 2: 5,8-Difluoroquinolin-2(1 H )-one

[0066] N -(2,5-Difluorophenyl)cinnamamide (8.00 g, 30.9 mmol) and solid aluminum chloride (6.10 g, 46.3 mmol) were stirred at 140 o °C for 2 h. The mixture was cooled to room temperature, water (20 mL) was added thereto, and the mixture was stirred for 1 h and then filtered. The filter cake was dried to obtain 5,8-difluoroquinolin-2(1 H )-one (6.00 g, 91.3% yield) as a red solid. LCMS calc. for C9H6F2NO [M+H] + : m / z = 182.0; Found: 182.1.

[0067] Step 3: 2-Chloro-5,8-difluoroquinoline

[0068] At room temperature, thionyl chloride H (7.88 g, 66.2 mmol) was added to a solution of 5,8-difluoroquinolin-2(1 )-one (6.00 g, 33.1 mmol) in DMF (10 mL). The reaction solution was stirred at 70 o °C for 1 h. After the reaction solution was cooled to room temperature, it was poured into water and neutralized to pH = 7 with an aqueous solution of sodium hydroxide (25%), and a solid precipitated. The solid was filtered and dried to obtain 2-chloro-5,8-difluoroquinoline (5.00 g, 64.5% yield). LCMS calc. for C9H5ClF2N [M+H] + : m / z = 200.0; Found: 200.1.

[0069] Step 4: tert-Butyl (5,8-difluoroquinolin-2-yl)carbamate

[0070] To a mixture of 2-chloro-5,8-difluoroquinoline (5.00 g, 25.1 mol), tert-butyl carbamate (5.88 g, 50.3 mmol), tris(dibenzylideneacetone)dipalladium (2.30 g, 2.51 mmol), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (2.90 g, 5.01 mmol) and cesium carbonate (16.4 g, 50.3 mmol) was added 1,4-dioxane (50 mL), and the reaction system was evacuated and replaced with nitrogen three times. The reaction solution was under nitrogen protection at 100 oStir for 16 h. The reaction solution was cooled to room temperature and filtered. The filtrate was concentrated and purified by silica gel column chromatography (ethyl acetate / petroleum ether = 20%) to obtain the solid tert-butyl (5,8-difluoroquinolin-2-yl)carbamate (5.50 g, 70.5% yield). LCMS calc. for C 14 H 15 F2N2O2[M+H] + : m / z = 281.1; Found: 281.1。

[0071] Step Five: 5,8-Difluoroquinolin-2-amine

[0072] To tert-butyl (5,8-difluoroquinolin-2-yl)carbamate (700 mg, 2.50 mmol) was added dioxane hydrochloride (4M) solution (10 mL), and the reaction solution was stirred at room temperature for 16 h. The reaction solution was concentrated to obtain the crude 5,8-difluoroquinolin-2-amine (500 mg), and the crude product was directly used in the next reaction. LCMS calc. for C9H7F2N2 [M+H] + : m / z =181.0; Found: 181.1。

[0073] Step Six: tert-Butyl ( S )-3-((5,8-difluoroquinolin-2-yl)carbamoyl)piperidine-1-carboxylate

[0074] To 5,8-difluoroquinolin-2-amine (1.80 g, 10.0 mmol), ( S )-1-(tert-butoxycarbonyl)piperidine-3-carboxylic acid (3.45 g, 15.1 mmol) in N , N -dimethylformamide (20 mL) solution was added HATU (5.70 g,15.1 mmol) and diisopropylethylamine (2.58 g, 20.0 mmol). The reaction solution was stirred at room temperature for 10 h. Saturated ammonium chloride aqueous solution (100 mL) was added to the reaction solution, and it was extracted with ethyl acetate (50 mL x 3). The combined organic phases were washed with saturated sodium chloride and dried over anhydrous sodium sulfate. The anhydrous sodium sulfate was filtered off, and the filtrate was concentrated and purified by silica gel column chromatography (ethyl acetate / petroleum ether = 20%) to obtain the solid tert-butyl ( S )-3-((5,8-difluoroquinolin-2-yl)carbamoyl)piperidine-1-carboxylate (1.00 g, 23% yield). LCMS calc. for C 20 H 24 F2N3O3 [M+H]+ : m / z = 392.2; Found: 392.1。

[0075] Step 7: ( S )- N -(5,8-Difluoroquinolin-2-yl)piperidine-3-carboxamide

[0076] At room temperature, trifluoroacetic acid (1.6 mL) was added to a solution of tert-butyl ( S )-3-((5,8-difluoroquinolin-2-yl)carbamoyl)piperidine-1-carboxylate (1.00 g, 2.55 mmol) in dichloromethane (8 mL), and the reaction mixture was stirred at room temperature for 3 hours. The reaction mixture was concentrated to obtain the crude product ( S )- N -(5,8-Difluoroquinolin-2-yl)piperidine-3-carboxamide (700 mg, 88.0% yield). LCMS calc. for C 15 H 16 F2N3O [M+H] + : m / z = 292.1; Found: 292.1。

[0077] Step 8: ( S )- N -(5,8-Difluoroquinolin-2-yl)-1-(1-methylazetidin-3-yl)piperidine-3-carboxamide

[0078] At room temperature, triethylamine (486 mg, 4.80 mmol) was added to a solution of 1-methylazetidin-3-one hydrochloride (351 mg, 2.88 mmol) in dichloromethane (2 mL), and the solution was stirred at room temperature for 10 minutes. Then, ( S )- N -(5,8-Difluoroquinolin-2-yl)piperidine-3-carboxamide (700 mg, 2.40 mmol) and titanium(IV) isopropoxide (342 mg, 1.20 mmol) were successively added to the reaction mixture. After stirring the reaction mixture at room temperature for 1 hour, sodium triacetoxyborohydride (1.02 g, 4.80 mmol) was added. The reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was filtered and concentrated, and then purified by preparative high performance liquid chromatography (column: XBridge C18 5μm 19 × 150 mm; mobile phase: ACN-H2O (0.1% TFA)) to obtain a white solid ( S )- N-(5,8-difluoroquinolin-2-yl)-1-(1-methylazetidin-3-yl)piperidine-3-carboxamide (27.8 mg). LCMS calc. for C 19 H 23 F2N4O [M+H] + : m / z = 361.2; Found: 361.3. 1 H NMR (400 MHz, DMSO- d 6) d 11.28 (s, 1H), 8.54 (d, J = 9.3 Hz, 1H), 8.46 (d, J = 8.9 Hz, 1H), 7.62-7.53(m, 1H), 7.31 (td, J = 9.1, 3.4 Hz, 1H), 4.44-4.29 (m, 2H), 4.12-3.79 (m, 5H), 3.24-2.91 (m, 3H), 2.87 (s, 3H), 2.04-1.94 (m, 1H), 1.87-1.70 (m, 1H), 1.66-1.42 (m, 2H).

[0079] Example 2 Preparation of Compound I-5

[0080] Step 1: N -(2,3,5-trifluorophenyl)cinnamamide

[0081] Dissolve 2,3,5-trifluoroaniline (4.41 g, 30.0 mmol) and potassium carbonate (8.29 g, 60.0 mmol) in water (30 mL) and acetonitrile (30 mL), and add cinnamoyl chloride (5.49 g, 33.0 mmol) dropwise at 0 °C. Stir the reaction mixture at 0 °C for 2 hours. Filter the reaction mixture, and dry the filter cake to obtain the product as a white solid N -(2,3,5-trifluorophenyl)cinnamamide (8.00 g). LCMS calc. for C 15 H 11 F3NO [M+H] + : m / z = 278.1; Found: 278.0.

[0082] Step 2: 5,7,8-trifluoroquinolin-2(1 H )-one

[0083] N -(2,3,5-Trifluorophenyl)cinnamamide (2.80 g, 10.1 mmol) and aluminum chloride (4.04 g, 30.3 mmol) were mixed and stirred at 140 °C for 1 hour. The reaction was cooled, water (20 mL) was added and stirred, and then filtered. The filter cake was slurried with acetonitrile to obtain 5,7,8-trifluoroquinolin-2(1 H )-one (1.20 g) as a brown solid. LCMS calc. for C9H5F3NO [M+H] + : m / z = 200.0; Found: 200.1.

[0084] Step 3: 2-Chloro-5,7,8-trifluoroquinoline

[0085] 5,7,8-Trifluoroquinolin-2(1 H )-one (1.20 g, 6.03 mmol) was dissolved in phosphorus oxychloride (10 mL), and the resulting solution was stirred at 80 °C for 1 hour. After the reaction solution was cooled to room temperature, it was poured into 100 mL of saturated sodium bicarbonate aqueous solution for quenching. It was extracted with ethyl acetate (50 mL x 3), the organic phases were combined, dried with anhydrous sodium sulfate and concentrated, and purified by silica gel chromatography (ethyl acetate / petroleum ether = 1:4) to obtain 2-chloro-5,7,8-trifluoroquinoline (0.66 g) as a brown solid. LCMS calc. for C9H4ClF3N [M+H] + : m / z = 218.0; Found: 217.9.

[0086] Step 4: tert-Butyl (5,7,8-trifluoroquinolin-2-yl)carbamate

[0087] 2-Chloro-5,7,8-trifluoroquinoline (0.66 g, 3.03 mmol), tert-butyl carbamate (0.43 g, 3.64 mmol), chloro[(4,5-bis(diphenylphosphino)-9,9-dimethylanthracene)-2-(2-aminobiphenyl)]palladium(II) (0.27 g, 0.30 mmol) and cesium carbonate (1.98 g, 6.07 mmol) were dissolved in dioxane (10 mL), and argon was evacuated and replaced three times. It was stirred at 80 °C for 2 hours in an argon atmosphere. The reaction solution was poured into 50 mL of water, extracted with ethyl acetate (20 mL x 3), the organic phases were combined, dried with anhydrous sodium sulfate and concentrated, and purified by silica gel chromatography (ethyl acetate / petroleum ether = 20%) to obtain tert-butyl (5,7,8-trifluoroquinolin-2-yl)carbamate (0.86 g) as a brown solid. LCMS calc. forC14 H 14 F3N2O2 [M+H] + : m / z = 299.1; Found: 242.9 [M-C4H7] + 。

[0088] Step Five: 5,7,8-Trifluoroquinolin-2-amine

[0089] Dissolve tert-butyl (5,7,8-trifluoroquinolin-2-yl)carbamate (0.40 g, 1.34 mmol) in dichloromethane (5 mL) and trifluoroacetic acid (5 mL), and stir at 20 °C for 0.5 h. Spin-dry the reaction solution, add 50 mL of saturated aqueous sodium bicarbonate, and extract with ethyl acetate (20 mL x 3). Dry the organic phase with anhydrous sodium sulfate and concentrate, and purify by silica gel chromatography (ethyl acetate / petroleum ether = 1:1) to obtain 5,7,8-trifluoroquinolin-2-amine (0.17 g) as a brown solid. LCMS calc. for C9H6F3N2 [M+H] + : m / z = 199.0; Found: 199.1.

[0090] Step Six: tert-Butyl ( S )-3-((5,7,8-trifluoroquinolin-2-yl)carbamoyl)piperidine-1-carboxylate

[0091] Dissolve 5,7,8-trifluoroquinolin-2-amine (170 mg, 0.86 mmol), 1-butylphosphonic anhydride (1.24 g, 50% ethyl acetate solution, 1.72 mmol), N , N -diisopropylethylamine (333 mg, 2.57 mmol) and ( S )-1-(tert-butoxycarbonyl)piperidine-3-carboxylic acid (295 mg, 1.29 mmol) in N , N -dimethylformamide (5 mL), and stir the reaction solution at 60 °C for 16 h. Dilute the reaction solution with 50 mL of water and extract with ethyl acetate (20 mL x 3). Combine the organic phases, dry with anhydrous sodium sulfate and concentrate, and purify by silica gel chromatography (petroleum ether / ethyl acetate = 3:1) to obtain tert-butyl ( S )-3-((5,7,8-trifluoroquinolin-2-yl)carbamoyl)piperidine-1-carboxylate (300 mg) as a white solid. LCMS calc. for C 20 H 23F3N3O3 [M+H] + : m / z = 410.4; Found: 410.1。

[0092] Step 7: ( S )- N -(5,7,8-Trifluoroquinolin-2-yl)piperidine-3-carboxamide

[0093] Dissolve tert-butyl ( S )-3-((5,7,8-trifluoroquinolin-2-yl)carbamoyl)piperidine-1-carboxylate (300 mg, 0.73 mmol) in 4M hydrochloric acid / ethyl acetate (5 mL), and stir the reaction mixture at 20 °C for 1 hour. Concentrate the reaction mixture to obtain a crude white solid ( S )- N -(5,7,8-Trifluoroquinolin-2-yl)piperidine-3-carboxamide (250 mg). LCMS calc. for C 15 H 15 F3N3O [M+H] + : m / z = 310.1; Found: 310.2。

[0094] Step 8: tert-Butyl ( S )-3-(3-((5,7,8-trifluoroquinolin-2-yl)carbamoyl)piperidin-1-yl)azetidine-1-carboxylate

[0095] Dissolve ( S )- N -(5,7,8-Trifluoroquinolin-2-yl)piperidine-3-carboxamide (250 mg, 0.81 mmol), tert-butyl 3-oxoazetidine-1-carboxylate (208 mg, 1.21 mmol) and acetic acid (0.09 mL, 1.62 mmol) in dichloromethane (10 mL), and add sodium triacetoxyborohydride (343 mg, 1.62 mmol) at 20 °C. Stir the reaction at 20 °C for 2 hours. Pour the reaction mixture into 50 mL of saturated aqueous sodium bicarbonate, and extract with dichloromethane (20 mL x 3). Combine the organic phases, dry over anhydrous sodium sulfate and concentrate, and purify by silica gel chromatography (petroleum ether / ethyl acetate = 1:1) to obtain the white solid tert-butyl ( S )-3-(3-((5,7,8-trifluoroquinolin-2-yl)carbamoyl)piperidin-1-yl)azetidine-1-carboxylate (300 mg). LCMS calc. for C 23 H 28 F3N4O3 [M+H]+ : m / z = 465.2; Found: 465.3。

[0096] Step Nine: ( S )-1-(azetidin-3-yl)- N -(5,7,8-trifluoroquinolin-2-yl)piperidine-3-carboxamide

[0097] Dissolve tert-butyl ( S )-3-(3-((5,7,8-trifluoroquinolin-2-yl)carbamoyl)piperidin-1-yl)azetidine-1-carboxylate (300 mg, 0.65 mmol) in dichloromethane (5 mL) and trifluoroacetic acid (5 mL), and stir the reaction at 20 °C for 0.5 h. Concentrate the reaction solution to obtain a crude colorless oil ( S )-1-(azetidin-3-yl)- N -(5,7,8-trifluoroquinolin-2-yl)piperidine-3-carboxamide (300 mg). LCMS calc. for C 18 H 20 F3N4O [M+H] + : m / z = 365.2; Found: 365.1。

[0098] Step Ten: ( S )-1-(1-methylazetidin-3-yl)- N -(5,7,8-trifluoroquinolin-2-yl)piperidine-3-carboxamide

[0099] Dissolve ( S )-1-(azetidin-3-yl)- N -(5,7,8-trifluoroquinolin-2-yl)piperidine-3-carboxamide (250 mg, 0.81 mmol), aqueous formaldehyde solution (247 mg, 30%, 2.47 mmol) and acetic acid (0.09 mL, 1.65 mmol) in methanol (10 mL), and add sodium cyanoborohydride (103 mg, 1.65 mmol) at 20 °C. Stir the reaction at 20 °C for 1 h. Pour the reaction solution into 50 mL of saturated aqueous sodium bicarbonate solution, and extract with ethyl acetate (20 mL x 3). Combine the organic phases, dry over anhydrous sodium sulfate and concentrate, and purify by preparative high performance liquid chromatography (column: XBridge C18 5μm 19 × 150 mm; mobile phase: ACN-H2O (0.1% NH4HCO3) gradient: 20% - 95%) to obtain a white solid ( S)-1-(1-Methylazetidin-3-yl)- N -(5,7,8-trifluoroquinolin-2-yl)piperidine-3-carboxamide (110 mg). LCMScalc. for C 19 H 22 F3N4O [M+H] + : m / z = 379.2; Found: 379.2. 1 H NMR (400 MHz, DMSO- d 6) d 11.36 (s, 1H), 8.54 – 8.40 (m, 2H), 7.68 (td, J J = 10.5, 5.9 Hz, 1H),3.43 – 3.38 (m, 2H), 2.88 – 2.68 (m, 5H), 2.56 – 2.52 (m, 1H), 2.22 (s, 3H),2.09 – 1.99 (m, 1H), 1.93 – 1.80 (m, 2H), 1.73 – 1.63 (m, 1H), 1.55 – 1.38(m, 2H).

[0100] The following compounds use the synthetic route method of Example 2 above or the corresponding intermediates of the modified above route method:

[0101] Example 3: Preparation of Compounds I-7, I-8

[0102] Step 1: (3 S )- N -(5,8-Difluoroquinolin-2-yl)-1-(1-methylpyrrolidin-3-yl)piperidine-3-carboxamide

[0103] ( S )- N-(5,8-Difluoroquinolin-2-yl)piperidine-3-carboxamide (291 mg, 1.00 mmol) was dissolved in dichloromethane (3 mL). 1-Methylpyrrolidin-3-one (197 mg, 1.99 mmol) and titanium tetraisopropoxide (1.42 mL, 0.50 mmol) were added, and the mixture was stirred for 10 minutes. Sodium triacetoxyborohydride (233 mg, 1.10 mmol) was added. The reaction mixture was stirred at 20 °C for 5 hours. The reaction mixture was poured into 50 mL of saturated aqueous sodium bicarbonate solution and extracted with dichloromethane (20 mL x 3). The combined organic phases were dried over anhydrous sodium sulfate and concentrated, and purified by silica gel chromatography (petroleum ether / ethyl acetate = 1:1) to obtain a white solid (3 S )- N -(5,8-Difluoroquinolin-2-yl)-1-(1-methylpyrrolidin-3-yl)piperidine-3-carboxamide (70 mg). LCMS calc. for C 20 H 25 F2N4O [M+H] + : m / z = 375.2; Found: 375.2。

[0104] Step 2: ( S )- N -(5,8-Difluoroquinolin-2-yl)-1-(( R )-1-methylpyrrolidin-3-yl)piperidine-3-carboxamide (I-7) and ( S )- N -(5,8-Difluoroquinolin-2-yl)-1-(( S )-1-methylpyrrolidin-3-yl)piperidine-3-carboxamide (I-8)

[0105] Chiral resolution of (3 S )- N -(5,8-Difluoroquinolin-2-yl)-1-(1-methylpyrrolidin-3-yl)piperidine-3-carboxamide (70 mg) was performed (column: CHIRALPAK AD-H, 250×20 mm I.D., 5 µm, mobile phase: 40% MeOH (NH4OH 0.2%):60% CO2, 40 mL / min, 100 bar) to obtain ( S )- N -(5,8-Difluoroquinolin-2-yl)-1-(( R )-1-methylpyrrolidin-3-yl)piperidine-3-carboxamide (I-7) (16.6 mg) (chiral analysis retention time: 1.12 min) and ( S)- N -(5,8-difluoroquinolin-2-yl)-1-(( S )-1-methylpyrrolidin-3-yl)piperidine-3-carboxamide (I-8) (13.2 mg) (Chiral analysis retention time: 1.25 min). Chiral analysis conditions (Chiral column: ChiralCelOJ-H, 500 × 4.6 mm I.D., 2µm; Methanol (NH4OH) in CO2).

[0106] ( S )- N -(5,8-difluoroquinolin-2-yl)-1-(( R )-1-methylpyrrolidin-3-yl)piperidine-3-carboxamide (I-7) analysis data

[0107] LCMS calc. for C 20 H 25 F2N4O [M+H] + : m / z = 375.2; Found: 375.3; 1 H NMR(400 MHz, CD3OD) d 8.49 - 8.44 (m, 2H), 7.42 - 7.35 (m, 1H), 7.18 - 7.10 (m, 1H), 3.76 - 3.55 (m, 4H), 3.48 - 3.34 (m, 1H), 3.23 - 3.05 (m, 2H), 2.98 - 2.81 (m, 5H), 2.80 - 2.69 (m, 1H), 2.50 - 2.40 (m, 1H), 2.35 - 2.25 (m, 1H), 2.10 - 2.02 (m, 1H), 1.98 - 1.92 (m, 1H), 1.85 - 1.75 (m, 2H).

[0108] ( S )- N -(5,8-difluoroquinolin-2-yl)-1-(( S )-1-methylpyrrolidin-3-yl)piperidine-3-carboxamide (I-8) analysis data

[0109] LCMS calc. for C 20 H 25 F2N4O [M+H] + : m / z = 375.2; Found: 375.3; 1 H NMR(400 MHz, CD3OD) d 8.50 - 8.40 (m, 2H), 7.38 - 7.32 (m, 1H), 7.13 - 7.08 (m, 1H), 3.22 - 3.17 (m, 1H), 3.15 - 3.07 (m, 1H), 3.06 - 2.85 (m, 3H), 2.83 - 2.75 (m, 1H), 2.74 - 2.66 (m, 2H), 2.58 (s, 3H), 2.52 - 2.44 (m, 1H), 2.37 - 2.30 (m, 1H), 2.19 - 2.10 (m, 1H), 2.01 - 1.89 (m, 2H), 1.87 - 1.76 (m, 1H), 1.74 - 1.61 (m, 2H).

[0110] Example 4 Preparation of Compounds I - 12 and I - 13

[0111] Step 1: 1 - (tert - butyl) 3 - methyl 5 - methoxypiperidine - 1,3 - dicarboxylate

[0112] Under an ice bath, sodium hydride (60% dispersed in mineral oil) (278 mg, 6.94 mmol) was added to a solution of methyl 1 - (tert - butyl) - 5 - hydroxy - 3 - piperidinecarboxylate (900 mg, 3.47 mmol) in N , N N,N - dimethylformamide (10 mL). After the reaction mixture was stirred at 0 °C for 1 hour, methyl iodide (985 mg, 6.94 mmol) was added dropwise, and the reaction was continued at room temperature for 1 hour. The reaction mixture was added to saturated ammonium chloride aqueous solution (100 mL) and extracted with ethyl acetate (30 mL × 3). The organic phase was dried over anhydrous sodium sulfate and concentrated to obtain the crude product 1 - (tert - butyl) 3 - methyl 5 - methoxypiperidine - 1,3 - dicarboxylate (900 mg). LCMS calc. for C 13 H 24 NO5 [M + H] + : m / z = 274.2; Found: 218.0 [M + H - C4H8] + .

[0113] Step 2: 1 - (tert - butoxycarbonyl) - 5 - methoxypiperidine - 3 - carboxylic acid

[0114] At room temperature, sodium hydroxide (391 mg, 9.77 mmol) was added to a solution of 1-(tert-butyl) 3-methyl 5-methoxypiperidine-1,3-dicarboxylate (890 mg, 3.26 mmol) in methanol (6 mL) and water (2 mL). The reaction mixture was stirred at 60 °C for 1 hour and then cooled to room temperature. The reaction mixture was adjusted to pH 5-6 with 1 M aqueous hydrochloric acid and extracted with ethyl acetate (30 mL x 3). The organic phase was dried over anhydrous sodium sulfate and concentrated to give the product 1-(tert-butoxycarbonyl)-5-methoxypiperidine-3-carboxylic acid (800 mg). LCMS calc. for C 12 H 22 NO5 [M+H] + : m / z = 260.1; Found:203.9 [M+H-C4H8] + .

[0115] Step 3: tert-Butyl 3-((5,8-difluoroquinolin-2-yl)carbamoyl)-5-methoxypiperidine-1-carboxylate

[0116] At room temperature, DIEA (1.18 g, 9.14 mmol) was added to a solution of 1-(tert-butoxycarbonyl)-5-methoxypiperidine-3-carboxylic acid (790 mg, 3.05 mmol), 5,8-difluoroquinolin-2-amine (549 mg, 3.05 mmol) and n-butylphosphoric anhydride (50% ethyl acetate solution) (3.29 g, 4.57 mmol) in N , N N,N-dimethylformamide (10 mL). The reaction mixture was stirred at 65 °C for 16 hours and then cooled to room temperature. The reaction mixture was poured into water (100 mL) and extracted with ethyl acetate (30 mL x 3). The organic phase was dried over anhydrous sodium sulfate and concentrated, and the residue was purified by silica gel chromatography (petroleum ether / ethyl acetate = 4:1) to give tert-Butyl 3-((5,8-difluoroquinolin-2-yl)carbamoyl)-5-methoxypiperidine-1-carboxylate (550 mg). LCMS calc. for C 21 H 26 F2N3O4 [M+H] + : m / z = 422.2; Found: 422.2.

[0117] Step 4: N -(5,8-Difluoroquinolin-2-yl)-5-hydroxypiperidine-3-carboxamide

[0118] A solution of tert-butyl 3-((5,8-difluoroquinolin-2-yl)carbamoyl)-5-methoxypiperidine-1-carboxylate (900 mg, 2.14 mmol) in dichloromethane (10 mL) was added dropwise with boron tribromide (2.68 g, 10.7 mmol) at 0 °C. After the reaction mixture was stirred at 20 °C for 8 hours, it was cooled to 0 °C and quenched with methanol (5 mL). The reaction mixture was concentrated, and the residue was purified by a C18 reverse-phase column (mobile phase A: water (0.1% ammonium bicarbonate), mobile phase B: acetonitrile, Gradient: 5% - 90% (%B)) to obtain N -(5,8-difluoroquinolin-2-yl)-5-hydroxypiperidine-3-carboxamide as a white solid (450 mg). LCMS calc. for C 15 H 16 F2N3O2 [M+H] + : m / z = 308.1; Found: 307.9.

[0119] Step 5: trans-tert-butyl 3-(3-((5,8-difluoroquinolin-2-yl)carbamoyl)-5-hydroxypiperidin-1-yl)azetidine-1-carboxylate and cis-tert-butyl 3-(3-((5,8-difluoroquinolin-2-yl)carbamoyl)-5-hydroxypiperidin-1-yl)azetidine-1-carboxylate

[0120] At room temperature, to N -(5,8-difluoroquinolin-2-yl)-5-hydroxypiperidine-3-carboxamide (350 mg, 1.14 mmol), tert-butyl 3-oxoazetidine-1-carboxylate (291 mg, 1.71 mmol) in dichloromethane (5 mL) and acetic acid (0.3 mL) was added sodium triacetoxyborohydride (483 mg, 2.28 mmol). After the reaction mixture was reacted at 20 °C for 2 hours, it was poured into water (10 mL), and extracted with dichloromethane (10 mL x 3). The organic phase was dried over anhydrous sodium sulfate and concentrated, and the residue was purified by silica gel chromatography (dichloromethane / methanol = 95:5) to obtain trans-tert-butyl 3-(3-((5,8-difluoroquinolin-2-yl)carbamoyl)-5-hydroxypiperidin-1-yl)azetidine-1-carboxylate (170 mg) (the component with less polarity on the TLC plate), cis-tert-butyl 3-(3-((5,8-difluoroquinolin-2-yl)carbamoyl)-5-hydroxypiperidin-1-yl)azetidine-1-carboxylate (220 mg) (the component with more polarity on the TLC plate). LCMS calc. for C 23 H 29F2N4O4 [M+H] + : m / z = 463.2; Found: 463.0。

[0121] Steps six to seven use cis-tert-butyl 3-(3-((5,8-difluoroquinolin-2-yl)carbamoyl)-5-hydroxypiperidin-1-yl)azetidine-1-carboxylate as the raw material and refer to the conditions of steps nine to ten in Example 2 to obtain cis- N -(5,8-difluoroquinolin-2-yl)-5-hydroxy-1-(1-methylazetidin-3-yl)piperidine-3-carboxamide

[0122] Step eight: (3 R ,5 S )- N -(5,8-difluoroquinolin-2-yl)-5-hydroxy-1-(1-methylazetidin-3-yl)piperidine-3-carboxamide (I-12) and (3 S ,5 R )- N -(5,8-difluoroquinolin-2-yl)-5-hydroxy-1-(1-methylazetidin-3-yl)piperidine-3-carboxamide (I-13)

[0123] Cis- N -(5,8-difluoroquinolin-2-yl)-5-hydroxy-1-(1-methylazetidin-3-yl)piperidine-3-carboxamide (230 mg, 0.61 mmol) is subjected to chiral resolution (instrument: SFC150, column: ChiralCel OJ-H, 250×30 mmI.D., 5 µm, mobile phase A: Supercritical CO2, mobile phase B: ETOH (0.1%NH 3· H2O) A:B = 70:30, 50 mL / min, 100 bar) to obtain a white solid (3 R ,5 S )- N -(5,8-difluoroquinolin-2-yl)-5-hydroxy-1-(1-methylazetidin-3-yl)piperidine-3-carboxamide (I-12) (112 mg) (chiral analysis retention time: 1.46min), (3 S ,5 R )- N-(5,8-Difluoroquinolin-2-yl)-5-hydroxy-1-(1-methylazetidin-3-yl)piperidine-3-carboxamide (I-13) (125 mg) (Retention time in chiral analysis: 2.88 min). Chiral analysis conditions (Chiral column: ChiralCel OJ-3, 500 × 4.6 mm I.D., 2 µm; 30% of ethanol (0.05% DEA) in CO2, 2.5 mL / min).

[0124] (3 R ,5 S )- N -Analysis data of (5,8-difluoroquinolin-2-yl)-5-hydroxy-1-(1-methylazetidin-3-yl)piperidine-3-carboxamide (I-12)

[0125] LCMS calc. for C 19 H 23 F2N4O2 [M+H] + : m / z = 377.2; Found: 377.2; 1 ​​​​​​​​​​​​​​​​​​​​​​​​​​​23 F2N4O2 [M+H] + : m / z = 377.2; Found: 377.2; 1 H NMR(400 MHz, DMSO- d 6) d 11.19 (s, 1H), 8.52 – 8.45 (m, 2H), 7.62 – 7.53 (m, 1H),7.34 – 7.25 (m, 1H), 4.83 (d, J = 5.2 Hz, 1H), 3.57 – 3.47 (m, 1H), 3.45 –3.39 (m, 2H), 2.91 – 2.74 (m, 6H), 2.23 (s, 3H), 2.09 – 2.02 (m, 1H), 1.80(t, J = 10.8 Hz, 1H), 1.52 (t, J = 9.4 Hz, 1H), 1.29 (dd, J = 12.0, 23.2 Hz,1H).

[0128] Example 5 Preparation of Compound I-14

[0129] Step 1: ( S )- N -(5,8-Difluoroquinolin-2-yl)-1-(1-(2-hydroxyethyl)azetidin-3-yl)piperidine-3-carboxamide

[0130] At room temperature, to ( S ) - 1-(Azetidin-3-yl)- N-(5,8-Difluoroquinolin-2-yl)piperidine-3-carboxamide (0.10 g, 0.29 mmol) and 1,4-dioxane-2,5-diol (69.0 mg, 0.58 mmol) were added to a mixed solution of dichloromethane (5 mL) and acetic acid (0.5 mL), and then sodium triacetoxyborohydride (183 mg, 0.87 mmol) was added. The reaction mixture was stirred at 35 °C for 15 minutes. The reaction was quenched with saturated sodium bicarbonate solution (10 mL), and the mixture was extracted with dichloromethane (30 mL × 3). The combined organic phases were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified successively by silica gel column chromatography (MeOH / DCM = 0% - 5%) and reverse-phase preparation (column: XBridge C18 5μm 19 150 mm; mobile phase: A (5 mM NH4HCO3 + 0.05% NH3H2O in Water) B (ACN)) to obtain a white solid ( S )- N -(5,8-Difluoroquinolin-2-yl)-1-(1-(2-hydroxyethyl)azetidin-3-yl)piperidine-3-carboxamide (20.0 mg, yield 17.7%). LCMS calc. for C 20 H 25 F2N4O2 [M+H] + : m / z = 391.2; Found: 391.2. 1 H NMR (400 MHz, DMSO- d 6) d 11.26 (br s, 1H), 8.64 – 8.31 (m, 2H), 7.61– 7.50 (m, 1H), 7.34 – 7.23 (m, 1H), 4.35 (br s, 1H), 3.45 – 3.34 (m, 4H),2.90 – 2.67 (m, 5H), 2.59 – 2.53 (m, 1H), 2.48 – 2.43 (m, 2H), 2.06 – 1.96(m, 1H), 1.92 – 1.78 (m, 2H), 1.71 – 1.63 (m, 1H), 1.55 – 1.40 (m, 2H).

[0131] Example 6 Preparation of Compound I-15

[0132] Step 1: ( S )- N -(5,8-difluoroquinolin-2-yl)-1-(1-(2-methoxyethyl)azetidin-3-yl)piperidine-3-carboxamide

[0133] Dissolve 1,1,2-trimethoxyethane (200 mg, 1.66 mmol) in trifluoroacetic acid (0.20 mL) and stir at 50 °C for 0.5 h. After cooling the reaction solution to room temperature, successively add methanol (5 mL) and (3 S )-1-(azetidin-3-yl)- N -(5,8-difluoroquinolin-2-yl)piperidine-3-carboxamide (200 mg, 0.58 mmol) to the reaction solution. Add sodium cyanoborohydride (72.57 mg, 1.15 mmol) to the reaction solution at 15 °C and stir for 2 h. Pour the reaction solution into 50 mL of saturated sodium bicarbonate aqueous solution and extract with ethyl acetate (20 mL x 3). Combine the organic phases, dry with anhydrous sodium sulfate and concentrate, and purify by high performance liquid chromatography (mobile phase: ACN-H2O (0.05% ammonia)) to obtain the product as a white solid( S )- N -(5,8-difluoroquinolin-2-yl)-1-(1-(2-methoxyethyl)azetidin-3-yl)piperidine-3-carboxamide (23.0 mg). LCMS calc. for C 21 H 27 F2N4O2 [M+H] + : m / z = 405.2, Found: 405.2. 1 H NMR (400 MHz, DMSO- d 6) d 11.27 (br s, 1H), 8.60 – 8.38 (m, 2H), 7.62 – 7.50 (m, 1H),7.29 (td, J = 9.1, 3.4 Hz, 1H), 3.44 – 3.38 (m, 2H), 3.29 – 3.25 (m, 2H),3.20 (s, 3H), 2.89 – 2.69 (m, 6H), 2.56 – 2.53 (m, 2H), 2.06 – 1.98 (m, 1H),1.89 – 1.80 (m, 2H), 1.71 – 1.62 (m, 1H), 1.56 – 1.44 (m, 2H).

[0134] Example 7 Preparation of Compounds I-16 and I-17

[0135] In Step 1 to Step 2, using the trans-tert-butyl 3-(3-((5,8-difluoroquinolin-2-yl)carbamoyl)-5-hydroxypiperidin-1-yl)azetidine-1-carboxylate obtained in Step 5 of Example 4 as the raw material, referring to the conditions of Step 9 to Step 10 of Example 2, trans- N -(5,8-Difluoroquinolin-2-yl)-5-hydroxy-1-(1-methylazetidin-3-yl)piperidine-3-carboxamide

[0136] Step 3: (3 R ,5 R )- N -(5,8-Difluoroquinolin-2-yl)-5-hydroxy-1-(1-methylazetidin-3-yl)piperidine-3-carboxamide (I-16) and (3 S ,5 S )- N -(5,8-Difluoroquinolin-2-yl)-5-hydroxy-1-(1-methylazetidin-3-yl)piperidine-3-carboxamide (I-17)

[0137] Trans- N -(5,8-Difluoroquinolin-2-yl)-5-hydroxy-1-(1-methylazetidin-3-yl)piperidine-3-carboxamide (100 mg, 0.27 mmol) was subjected to chiral resolution (instrument: SFC150, column: ChiralCel AD-H, 250×30 mm I.D., 5 µm, mobile phase A: Supercritical CO2, mobile phase B: EtOH (0.1% NH3H2O) A:B = 50:50, 50 mL / min, 100 bar) to obtain a white solid product (3 R ,5 R )- N -(5,8-Difluoroquinolin-2-yl)-5-hydroxy-1-(1-methylazetidin-3-yl)piperidine-3-carboxamide (I-16) (48 mg) (chiral analysis retention time: 0.40 min), (3 S ,5 S )- N-(5,8-Difluoroquinolin-2-yl)-5-hydroxy-1-(1-methylazetidin-3-yl)piperidine-3-carboxamide (I-17) (38 mg) (Retention time in chiral analysis: 3.65 min). Chiral analysis conditions (Chiral column: ChiralCel AD-3, 500 × 4.6 mm I.D., 2 µm; 40% of ethanol (0.05% DEA) in CO2, 2.5 mL / min).

[0138] (3 R ,5 R )- N -(5,8-Difluoroquinolin-2-yl)-5-hydroxy-1-(1-methylazetidin-3-yl)piperidine-3-carboxamide (I-16) analysis data

[0139] LCMS calc. for C 19 H 23 F2N4O2 [M+H] + : m / z = 377.2; Found: 377.3; 1 H NMR(400 MHz, DMSO- d 6) d 11.35 (s, 1H), 8.50 – 8.46 (m, 2H), 7.60 – 7.50 (m, 1H),7.35 – 7.25 (m, 1H), 4.63 (s, 1H), 3.85 (s, 1H), 3.52– 3.40 (m, 3H), 2.98 –2.87 (m, 4H), 2.43 – 2.40 (m, 2H), 2.30 (s, 3H), 2.10 – 2.03 (m, 1H), 1.97 –1.84 (m, 1H), 1.63 – 1.49 (m, 1H), (3 S ,5 S )- N -(5,8-Difluoroquinolin-2-yl)-5-hydroxy-1-(1-methylazetidin-3-yl)piperidine-3-carboxamide (I-17) analysis data LCMS calc. for C 19 H 23 F2N4O2 [M+H] + : m / z = 377.2; Found: 377.3; 1 H NMR(400 MHz, DMSO- d 6) d 11.37 (s, 1H), 8.50 – 8.49 (m, 2H), 7.58 – 7.52 (m, 1H),7.32 – 7.25 (m, 1H), 4.62 (d, J = 4.4 Hz, 1H), 3.88 - 3.79 (m, 1H), 3.49 –3.39 (m, 3H), 3.00 – 2.76 (m, 4H), 2.45 – 2.24 (m, 2H), 2.24 (s, 3H), 2.08 –2.00 (m, 1H), 1.96 – 1.85 (m, 1H), 1.61 – 1.51 (m, 1H).

[0140] Example 8 Preparation of Compound I-19

[0141] Step 1: 3-Benzyl 1-(tert-butyl) ( S )-piperidine-1,3-dicarboxylate

[0142] At room temperature, potassium carbonate (90.5 g, 0.66 mol) and benzyl bromide (82 g, 0.48 mol) were added to a solution of ( S )-1-(tert-butoxycarbonyl)piperidine-3-carboxylic acid (100 g, 0.44 mol) in N , N -dimethylformamide (500 mL). After stirring the reaction mixture at room temperature for 3 hours, water (2500 mL) was added, and the mixture was extracted with ethyl acetate (1000 mL × 3). The combined organic phases were washed with saturated sodium chloride (800 mL × 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain 3-benzyl 1-(tert-butyl) ( S )-piperidine-1,3-dicarboxylate (139 g) as a pale yellow liquid.

[0143] Step 2: Benzyl ( S )-piperidine-3-carboxylate

[0144] At 0 °C, a solution of hydrochloric acid in ethyl acetate (700 mL, 5 mol / L) was added to a solution of 3-benzyl 1-(tert-butyl) ( S )-piperidine-1,3-dicarboxylate (139 g) in ethyl acetate (700 mL). After stirring the reaction mixture at room temperature for 1 hour, it was concentrated to obtain benzyl ( S )-piperidine-3-carboxylate hydrochloride (107 g).

[0145] Step 3: Benzyl ( S )-1-(1-(tert-butoxycarbonyl)azetidin-3-yl)piperidine-3-carboxylate

[0146] At 0 °C, sodium triacetoxyborohydride (112.3 g, 0.53 mmol) was added to a solution of benzyl ( S )-piperidine-3-carboxylate hydrochloride (89.6 g, 0.35 mol) and tert-butyl 3-oxoazetidine-1-carboxylate (90.7 g, 0.53 mmol) in tetrahydrofuran (500 mL). After the reaction mixture was warmed to room temperature and stirred for 2 hours, water (1500 mL) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (600 mL × 3). The combined organic phases were washed with saturated brine (500 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain crude benzyl ( S )-1-(1-(tert-butoxycarbonyl)azetidin-3-yl)piperidine-3-carboxylate (132 g).

[0147] Step 4: Benzyl ( S )-1-(azetidin-3-yl)piperidine-3-carboxylate

[0148] At room temperature, hydrochloric acid (600 mL, 5 M in EtOAc) was added to a solution of benzyl ( S )-1-(1-(tert-butoxycarbonyl)azetidin-3-yl)piperidine-3-carboxylate (132 g, 0.35 mol) in ethyl acetate (600 mL). After the reaction mixture was stirred at room temperature for 1 hour, the reaction mixture was concentrated to obtain the hydrochloride salt of benzyl ( S )-1-(azetidin-3-yl)piperidine-3-carboxylate (122.4 g) as a white solid.

[0149] Step 5: Benzyl ( S )-1-(1-methylazetidin-3-yl)piperidine-3-carboxylate

[0150] To benzyl ( S)-1-(Azetidin-3-yl)piperidine-3-carboxylate hydrochloride (118 g, 0.34 mmol) in tetrahydrofuran (600 mL) was added with 37% aqueous formaldehyde solution (143 g). At 0 °C, sodium triacetoxyborohydride (297 g, 1.4 mmol) was added to the reaction solution and stirred for 15 minutes, then the reaction solution was warmed to room temperature and stirred for 2 hours. Water (1000 mL) was added to the reaction solution, and it was extracted with dichloromethane (1000 mL x 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain a yellow oil, benzyl ( S )-1-(1-Methylazetidin-3-yl)piperidine-3-carboxylate (82 g).

[0151] Step 6: ( S )-1-(1-Methylazetidin-3-yl)piperidine-3-carboxylic acid

[0152] To a solution of benzyl ( S )-1-(1-Methylazetidin-3-yl)piperidine-3-carboxylate (82 g, 0.28 mol) in methanol (800 mL) was added 10% palladium on carbon (8 g), and the hydrogen was evacuated and replaced three times. The reaction solution was stirred at 60 °C overnight under a hydrogen pressure of 1 atm. The reaction solution was cooled to room temperature, the palladium on carbon was filtered off, and the filtrate was concentrated to obtain a solid ( S )-1-(1-Methylazetidin-3-yl)piperidine-3-carboxylic acid (55 g). 1 H NMR (400 MHz, DMSO- d 6) d 3.51 – 3.47 (m, 2H), 2.97 –2.75 (m, 3H), 2.75 – 2.58 (m, 1H), 2.50 – 2.46 (m, 1H), 2.40 – 2.31 (m, 1H),2.29 (s, 3H), 1.94 – 1.90 (m, 1H), 1.81 – 1.76 (m, 2H), 1.69 – 1.56 (m, 1H),1.48 – 1.28 (m, 2H).

[0153] Steps 7 to 12 were carried out according to the conditions of Steps 1 to 6 of Example 1, and the product ( S )- N -(5-Chloro-8-fluoroquinolin-2-yl)-1-(1-methylazetidin-3-yl)piperidine-3-carboxamide (I-19) was obtained from the starting material 5-chloro-2-fluoroaniline. LCMS calc. for C 19 H 23ClFN4O [M+H] + : m / z = 377.2; Found: 376.9; 1 H NMR (400 MHz, DMSO- d 6) d 11.29 (s, 1H), 8.58 – 8.51 (m, 2H), 7.65 – 7.54 (m, 2H), 3.51 - 3.50 (m, 2H), 2.96 - 2.90 (m, 3H), 2.82 – 2.74 (m, 2H), 2.56 – 2.54 (m, 1H), 2.31 (s, 3H), 2.03 (t, J = 9.6 Hz, 1H), 1.88 – 1.86 (m, 2H), 1.72 – 1.66 (m, 1H), 1.54 – 1.41 (m, 2H).

[0154] Example 9 Preparation of Compounds I-20-P1 and I-20-P2

[0155] Steps 1 to 5 refer to the conditions of Steps 6 to 10 in Example 7 to obtain N -(5,8-difluoroquinolin-2-yl)-4,4-dimethyl-1-(1-methylazetidin-3-yl)pyrrolidine-3-carboxamide (200 mg) was subjected to chiral resolution (column: CHIRALPAK AD-H 250 mm × 20 mm, 5 µm; mobile phase: 40% MeOH (NH4OH, 0.2%): 60% CO2; flow rate: 40 g / min) to obtain ( R )- N -(5,8-difluoroquinolin-2-yl)-4,4-dimethyl-1-(1-methylazetidin-3-yl)pyrrolidine-3-carboxamide (I-20-P1) (48.6 mg) (chiral analysis retention time: 1.81 min) and ( S )-( N -(5,8-difluoroquinolin-2-yl)-4,4-dimethyl-1-(1-methylazetidin-3-yl)pyrrolidine-3-carboxamide (I-20-P2) (45.8 mg) (chiral analysis retention time: 2.09 min). Chiral analysis conditions (chiral column: CHIRALPAK AD-H 250 mm × 4.6 mm, 5 µm; MeOH (NH4OH) in CO2).

[0156] I-20-P1 Analysis Data: LCMS calc. for C 20 H 25 F2N4O [M+H] + : m / z = 375.2; Found: 375.1. 1 H NMR (400 MHz, DMSO- d 6 ) d 11.13 (s, 1H), 8.50 (s, 2H), 7.56 (ddd, J = 10.7, 8.7, 4.7 Hz, 1H), 7.29 (td, J = 9.1, 3.5 Hz, 1H), 3.33 (t, J = 6.8 Hz, 2H), 3.16 – 3.07 (m, 1H), 2.99 (t, J = 7.1 Hz, 1H), 2.93 – 2.82 (m, 3H), 2.78(dd, J = 9.2, 6.6 Hz, 1H), 2.41 (d, J = 8.4 Hz, 1H), 2.29 – 2.18 (m, 4H), 1.23 (s, 3H), 0.98 (s, 3H).

[0157] I-20-P2 Analysis Data: LCMS calc. for C 20 H 25 F2N4O [M+H] + : m / z = 375.2; Found: 375.1. 1 H NMR (400 MHz, DMSO- d 6 ) d 11.13 (s, 1H), 8.50 (s, 2H), 7.56 (ddd, J = 10.7, 8.7, 4.7 Hz, 1H), 7.29 (td, J = 9.1, 3.4 Hz, 1H), 3.33 (t, J = 6.6 Hz, 2H), 3.17 – 3.06 (m, 1H), 2.99 (t, J = 7.1 Hz, 1H), 2.94 – 2.84 (m, 3H), 2.78(dd,J = 9.2, 6.6 Hz, 1H), 2.41 (d, J = 8.4 Hz, 1H), 2.32 – 2.16 (m, 4H),1.23 (s, 3H), 0.99 (s, 3H).

[0158] Example 10 Preparation of Compounds I-22-P1 and I-22-P2

[0159] Step 1: trans-1-Benzyl-4-methylpyrrolidine-3-carboxylic acid

[0160] LiOH (1.23 g, 51.5 mmol) was added to a solution of methyl trans-1-benzyl-4-methylpyrrolidine-3-carboxylate (4.00 g, 17.2 mmol) in methanol (40 mL) and water (10 mL). The mixture was stirred at 25 °C for 2 h. 2M aqueous HCl solution was slowly added to the reaction mixture until pH = 2. The residue was purified by reverse preparative HPLC (column: Gemini-C18 150 x 21.2 mm, 5 μm; mobile phase: ACN-H2O (0.1% FA); gradient: 5% - 20%) to give trans-1-benzyl-4-methylpyrrolidine-3-carboxylic acid as a white solid (3.00 g, yield 79.8%). LCMS calc. for C 13 H 18 NO2 [M+H] + : m / z = 220.1; Found: 220.1。

[0161] Step 2: trans-1-Benzyl- N -(5,8-difluoroquinolin-2-yl)-4-methylpyrrolidine-3-carboxamide

[0162] trans-1-Benzyl-4-methylpyrrolidine-3-carboxylic acid (3.00 g, 13.7 mmol) was dissolved in pyridine (30 mL), and 5,8-difluoroquinolin-2-amine (4.45 g, 20.5 mmol) and T4P (29.6 g, 41.1 mmol, 50% in EtOAc) were added. The mixed solution was stirred at 40 °C for 18 h under nitrogen protection. Water (50 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (70 mL x 3), washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (0 - 60% ethyl acetate in petroleum ether) to give trans-1-benzyl-N -(5,8-Difluoroquinolin-2-yl)-4-methylpyrrolidine-3-carboxamide (2.20 g, yield 42.2%). LCMS calc. for C 22 H 22 F2N3O [M+H] + : m / z = 382.2; Found: 382.0。

[0163] Step 3: trans- N -(5,8-Difluoroquinolin-2-yl)-4-methylpyrrolidine-3-carboxamide

[0164] trans - 1-Benzyl- N -(5,8-Difluoroquinolin-2-yl)-4-methylpyrrolidine-3-carboxamide (2.20 g, 5.77 mmol) in methanol (30 mL) was added with 10% palladium on carbon (1.84 g, 17.3 mmol). The mixture was stirred at 25 °C for 16 h under H2 atmosphere. After the reaction solution was filtered and rinsed with methanol, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (0 - 15% methanol in dichloromethane) to obtain the white solid trans -N -(5,8-Difluoroquinolin-2-yl)-4-methylpyrrolidine-3-carboxamide (1.20 g, yield 89.5%). LCMS calc. for C 15 H 16 F2N3O [M+H] + : m / z = 292.1; Found: 292.0。

[0165] Step 4: trans-tert-Butyl 3-(3-((5,8-difluoroquinolin-2-yl)carbamoyl)-4-methylpyrrolidin-1-yl)azetidine-1-carboxylate

[0166] trans -NA solution of (5,8-difluoroquinolin-2-yl)-4-methylpyrrolidine-3-carboxamide (1.20 g, 4.12 mmol) in DCM (15 mL) was added with tert-butyl 3-oxoazetidine-1-carboxylate (1.06 g, 6.18 mmol) and acetic acid (0.248 g, 4.12 mmol). After the mixture was stirred at 25 °C for 0.5 h, sodium triacetoxyborohydride (2.18 g, 10.3 mmol) was added, and the mixture was stirred at 25 °C for 2 h. Saturated aqueous sodium bicarbonate solution (50 mL) was added to the reaction system, and the mixture was extracted with ethyl acetate (60 mL × 3). The extract was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (0 - 60% ethyl acetate in petroleum ether) to obtain trans-tert-butyl 3-(3-((5,8-difluoroquinolin-2-yl)carbamoyl)-4-methylpyrrolidin-1-yl)azetidine-1-carboxylate (1.10 g, yield 59.8%). LCMS calc. for C 23 H 29 F2N4O3 [M+H] + : m / z = 447.2; Found: 447.0。

[0167] Step 5: trans-1-(Azetidin-3-yl)-N-(5,8-difluoroquinolin-2-yl)-4-methylpyrrolidine-3-carboxamide

[0168] Trans-tert-butyl 3-(3-((5,8-difluoroquinolin-2-yl)carbamoyl)-4-methylpyrrolidin-1-yl)azetidine-1-carboxylate (300 mg, 0.67 mmol) was dissolved in DCM (10 mL) and TFA (2 mL), and the mixture was stirred at 25 °C for 2 h. The mixture was concentrated under reduced pressure to obtain trans-1-(azetidin-3-yl)-N-(5,8-difluoroquinolin-2-yl)-4-methylpyrrolidine-3-carboxamide (210 mg, yield 90.3%). LCMS calc. for C 18 H 21 F2N4O [M+H] + : m / z = 347.2;Found: 347.0。

[0169] Step 6: (3 R ,4 R )- N -(5,8-difluoroquinolin-2-yl)-4-methyl-1-(1-methylazetidin-3-yl)pyrrolidine-3-carboxamide (I-22-P1) and (3 S ,4 S )-N -(5,8-Difluoroquinolin-2-yl)-4-methyl-1-(1-methylazetidin-3-yl)pyrrolidine-3-carboxamide (I-22-P2)

[0170] To a solution of trans-1-(azetidin-3-yl)-N-(5,8-difluoroquinolin-2-yl)-4-methylpyrrolidine-3-carboxamide (210 mg, 0.72 mmol) in DCM (5 mL) was added aqueous formaldehyde solution (93.7 mg, 1.08 mmol), acetic acid (43.3 mg, 0.72 mmol). After the mixture was stirred at 25 °C for 0.5 h, sodium triacetoxyborohydride (382 mg, 1.80 mmol) was added, and the mixture was stirred at 25 °C for 2 h. Saturated aqueous sodium bicarbonate solution (20 mL) was added to the reaction system, and the mixture was extracted with dichloromethane / methanol (10 / 1, 50 mL x 3), concentrated under reduced pressure, and the residue was purified by preparative reversed-phase HPLC (column: Gemini-C18 150 x  21.2 mm, 5 μm; mobile phase: ACN-H2O (0.1% FA); gradient: 5% - 50%) to obtain the white solid trans- N -(5,8-Difluoroquinolin-2-yl)-4-methyl-1-(1-methylazetidin-3-yl)pyrrolidine-3-carboxamide (150 mg, yield 57.7%). LCMS calc. for C 19 H 23 F2N4O [M+H] + : m / z = 361.2; Found: 361.0。

[0171] Trans- N -(5,8-Difluoroquinolin-2-yl)-4-methyl-1-(1-methylazetidin-3-yl)pyrrolidine-3-carboxamide (150 mg) was subjected to chiral resolution (column: CHIRALPAK AD-H 250 mm × 20 mm, 5 µm; mobile phase: 40% MeOH (NH4OH, 0.2%): 60% CO2; flow rate: 40 g / min) to obtain (3 R ,4 R )- N -(5,8-Difluoroquinolin-2-yl)-4-methyl-1-(1-methylazetidin-3-yl)pyrrolidine-3-carboxamide (I-22-P1) (63.3 mg) (chiral analysis retention time: 2.66 min). And (3 S ,4 S )- N-(5,8-Difluoroquinolin-2-yl)-4-methyl-1-(1-methylazetidin-3-yl)pyrrolidine-3-carboxamide (I-22-P2) (69.8 mg) (Retention time in chiral analysis: 3.66 min). Chiral analysis conditions (Chiral column: CHIRALPAK AD-H 250 mm × 4.6 mm, 5 µm; MeOH (NH4OH) in CO2).

[0172] Analytical data of I-22-P1: LCMS calc. for C 19 H 23 F2N4O [M+H] + : m / z = 361.2; Found: 361.0. 1 H NMR (400 MHz, CD3OD) d 8.48 (dt, J = 7.4, 5.1 Hz, 2H), 7.40 - 7.35 (m, 1H), 7.16 - 7.09 (m, 1H), 3.72 - 3.61 (m, 2H), 3.(此处原文可能有误,推测为3.30)30 - 3.21 (m, 3H), 3.00 - 2.90 (m, 2H), 2.85 (t, J = 8.8 Hz, 1H), 2.78 (dd, J = 13.6, 6.7 Hz, 1H), 2.62 (dt, J = 14.0, 6.9 Hz, 1H), 2.48 (s, 3H), 2.23 - 2.16 (m, 1H), 1.19 (d, J = 6.8 Hz, 3H).

[0173] Analytical data of I-22-P2: LCMS calc. for C 19 H 23 F2N4O [M+H] + : m / z = 361.2; Found: 361.0. 1 H NMR (400 MHz, CD3OD) d 8.55 - 8.45 (m, 2H), 7.38 (td, J = 10.1, 4.6 Hz, 1H), 7.14 (td, J = 9.0, 3.2 Hz, 1H), 3.92 (s, 2H), 3.60 (dd, J= 15.4, 9.3 Hz, 2H), 3.37 (dd, J = 12.6, 6.3 Hz, 1H), 3.03 - 2.97 (m, 2H), 2.90 - 2.75 (m, 2H), 2.69 (s, 3H), 2.63 (dt, J = 13.6, 6.9 Hz, 1H), 2.20 (t, J = 8.3 Hz, 1H), 1.20 (d, J = 6.8 Hz, 3H).

[0174] Example 11 Preparation of Compound I - 23 (P1 - 1, P1 - 2, P2 - 1 and P2 - 2)

[0175] Note: “or 1”, “or 2”, “or 3” and “or 4” in the structural formula indicate that the compound is in a single stereoconfiguration, but its configuration is uncertain.

[0176] Step 1: 1 - (tert - butyl) 3 - methyl 6 - oxopiperidine - 1,3 - dicarboxylate

[0177] Dissolve methyl 6 - oxopiperidine - 3 - carboxylate (5.0 g, 31.8 mmol) and DMAP (1.94 g, 15.9 mmol) in acetonitrile (50 mL), and slowly add (Boc)2O (10.4 g, 47.7 mmol). Stir the reaction mixture at 20 o °C for 12 hours, concentrate under reduced pressure, add water (10 mL), adjust the pH to 3 with 1 M aqueous NaHSO4 solution, extract with ethyl acetate (50 mL x 3). Combine the organic phases, dry over anhydrous sodium sulfate and concentrate to obtain the yellow solid 1 - (tert - butyl) 3 - methyl 6 - oxopiperidine - 1,3 - dicarboxylate (7.2 g). LCMS calc. for C 12 H 20 NO5 [M + H] + : m / z = 258.1; Found: 158.1 [M - C5H8O2 + H] + .

[0178] Step 2: 1 - (tert - butyl) 3 - methyl 6 - hydroxypiperidine - 1,3 - dicarboxylate

[0179] Dissolve 1-(tert-butyl) 3-methyl 6-oxopiperidine-1,3-dicarboxylate (5 g, 19.4 mmol) in tetrahydrofuran (130 mL), cool to -80 o °C, and slowly add dropwise DIBAL-H (24.3 mL, 24.3 mmol, 1 M in hexane). After the addition is complete, stir the reaction mixture at -80 o °C for 1 hour. Quench the reaction by adding saturated ammonium chloride (33 mL) (control the temperature < 10 o °C), stir for 15 minutes, then quickly filter, separate the layers, concentrate the organic phase, add water (20 mL), extract with diethyl ether (50 mL x 2). Combine the organic phases, dry over anhydrous sodium sulfate and concentrate to obtain 1-(tert-butyl) 3-methyl 6-hydroxypiperidine-1,3-dicarboxylate as a yellow oil (5.30 g crude product), which is directly used in the next step.

[0180] Step 3: 1-(tert-butyl) 3-methyl 3,4-dihydropyridine-1,3(2 H )-dicarboxylate

[0181] Dissolve 1-(tert-butyl) 3-methyl 6-hydroxypiperidine-1,3-dicarboxylate (5.30 g, 10.2 mmol) in anhydrous toluene (53 mL), cool to -80 o °C, and slowly add dropwise TFAA (2.14 g, 10.2 mmol). After the addition is complete, stir for 20 minutes. At -80 o °C, slowly add dropwise DIEA (5.01 mL, 38.8 mmol) to the reaction mixture. Let the reaction mixture warm naturally from -80 o °C to room temperature and stir for 12 hours. Cool to 0 o °C, add water (8 mL) to quench, adjust the pH to 6 with 10% citric acid (aq.), extract with ethyl acetate (50 mL × 3). Combine the organic phases, dry over anhydrous sodium sulfate and concentrate. Purify by silica gel chromatography (ethyl acetate / petroleum ether = 1:8) to obtain 1-(tert-butyl) 3-methyl 3,4-dihydropyridine-1,3(2 H )-dicarboxylate as a pale yellow oil (1.85 g). LCMS calc. for C 12 H 20 NO4 [M+H] + : m / z = 242.1; Found: 142.1 [M-C5H8O2+H] + .

[0182] Step 4: 2-(tert-Butyl) 4-methyl 2-azabicyclo[4.1.0]heptane-2,4-dicarboxylate

[0183] - 40 o At -40 °C, diethylzinc (24.7 mL, 24.7 mmol, 1 M n-hexane solution) was added to a solution of 1-(tert-butyl) 3-methyl 3,4-dihydropyridine-1,3(2H)-dicarboxylate (2.40 g, 9.9 mmol) in anhydrous dichloromethane (50 mL). After the addition, the mixture was stirred for 5 minutes, and then cooled to -55 o °C. Iodochloromethane (4.47 g, 25.3 mmol) was added, and the reaction mixture was allowed to warm to room temperature and stirred for 12 hours at -55 o °C. The reaction mixture was cooled to 0 o °C, quenched with saturated aqueous sodium bicarbonate (200 mL), stirred for 10 minutes, filtered, and extracted with dichloromethane (200 mL x 3). The combined organic phases were dried over anhydrous sodium sulfate and concentrated to give a brown solid, 2-(tert-butyl) 4-methyl 2-azabicyclo[4.1.0]heptane-2,4-dicarboxylate (1.1 g crude). LCMS calc. for C 13 H 22 NO4 [M+H] + : m / z = 256.1; Found: 200.1 [M-C4H8+H] + .

[0184] Step 5: 2-(tert-Butoxycarbonyl)-2-azabicyclo[4.1.0]heptane-4-carboxylic acid

[0185] 2-(tert-Butyl) 4-methyl 2-azabicyclo[4.1.0]heptane-2,4-dicarboxylate (1.17 g, 4.6 mmol) was dissolved in methanol (65 mL), and sodium hydroxide (0.50 g, 12.4 mmol) and water (6.5 mL) were added. The mixture was stirred at 20 o °C for 24 hours. The reaction mixture was concentrated under reduced pressure, 40 mL of aqueous solution was added, and the unreacted starting materials were extracted with diethyl ether (20 mL x 3). The aqueous phase was adjusted to pH = 2 with 1 M NaHSO4(aq.), extracted with chloroform (20 mL x 3), and the organic phase was dried over anhydrous sodium sulfate and concentrated to give a brown solid, 2-(tert-butoxycarbonyl)-2-azabicyclo[4.1.0]heptane-4-carboxylic acid (0.80 g crude). LCMS calc. for C 12 H 20 NO4 [M+H]+ : m / z = 242.1; Found: 186.1 [M-C4H8+H] + .

[0186] Step 6: tert-Butyl-4-((5,8-difluoroquinolin-2-yl)carbamoyl)-2-azabicyclo[4.1.0]heptane-2-carboxylic acid ester

[0187] 5,8-Difluoroquinoline-2-amine (420 mg, 2.33 mmol), 1-butylphosphine anhydride (5.04 g, 50% ethyl acetate solution, 6.99 mmol), and 2-(tert-butoxycarbonyl)-2-azabicyclo[4.1.0]heptane-4-carboxylic acid (791 mg, 3.26 mmol) were dissolved in pyridine (3 mL), and the reaction solution was heated to 20 °C. o Stirred at C for 3 hours. Add water (10 mL) and extract with ethyl acetate (10 mL x 3). Combine the organic phases, dry with anhydrous sodium sulfate and concentrate. Analyze by silica gel chromatography (petroleum ether / ethyl acetate = 3:1) to give tert-butyl-4-((5,8-difluoroquinolin-2-yl)carbamoyl)-2-azabicyclo[4.1.0]heptane-2-carboxylic acid ester (P1: 125 mg (LC-MS retention time 1.37 min)) and (P2: 65 mg (LC-MS retention time 1.39 min)). 21 H 24 F2N3O3 [M+H + : m / z = 404.2; Found:404.1.

[0188] Step 10: N -(5,8-difluoroquinolin-2-yl)-2-(1-methylacetidin-3-yl)-2-azabicyclo[4.1.0]heptane-4-carboxamide (I-23 P1-1 and P1-2)

[0189] Steps seven through ten were performed under the same conditions as in Example 7, using tert-butyl-4-((5,8-difluoroquinolin-2-yl)carbamoyl)-2-azabicyclo[4.1.0]heptane-2-carboxylic acid ester (P1: 125 mg (LC-MS retention time 1.37 min)) as the starting material to obtain N -(5,8-difluoroquinolin-2-yl)-2-(1-methylacetidin-3-yl)-2-azabicyclo[4.1.0]heptane-4-carboxamide (65 mg). The obtained N-(5,8-difluoroquinolin-2-yl)-2-(1-methylacetidin-3-yl)-2-azabicyclo[4.1.0]heptane-4-carboxamide (65 mg) was chirally resolved (column: CHIRALPAK AD-H, 250 mm × 20 mm, 5 µm; mobile phase: 40% MeOH (NH4OH, 0.2%): 60% CO2, 40 mL / min, 100 bar) to give fraction P1-1 (20.8 mg) of compound I-23 (chiral analysis retention time: 2.48 min) and fraction P1-2 (23.3 mg) of compound I-23 (chiral analysis retention time: 3.44 min). Chiral analysis conditions (chiral column: CHIRALPAK AD-H 250 mm × 4.6 mm, 5 µm; Methanol (NH4OH) in CO2).

[0190] Component P1-1 analysis data of compound I-23

[0191] LCMS calc. for C 20 H 23 F2N4O [M+H] + m / z = 373.2; Found: 373.2; 1 H NMR (400 MHz, MeOD) d 8.50 (d, J = 9.2 Hz, 1H), 8.46 (d, J = 9.2 Hz, 1H), 7.38(td, J = 9.6, 4.6 Hz, 1H), 7.13 (td, J = 9.0, 3.4 Hz, 1H), 3.63 (q, J = 6.5Hz, 2H), 3.43 – 3.36 (m, 1H), 3.20 (t, J = 7.0 Hz, 1H), 3.11 (t, J = 7.0 Hz,1H), 2.68 – 2.55 (m, 2H), 2.50 – 2.44 (m, 1H), 2.43 (s, 3H), 2.37 – 2.31 (m,1H), 2.30 – 2.21 (m, 1H), 2.01 (dd, J= 14.0, 4.2 Hz, 1H), 1.27 – 1.16 (m,1H), 0.54 – 0.42 (m, 2H).

[0192] Component P1-2 analysis data of compound I-23

[0193] LCMS calc. for C 20 H 23 F2N4O [M+H] + m / z = 373.2; Found: 373.2; 1 H NMR (400 MHz, MeOD) d 8.50 (d, J = 9.2 Hz, 1H), 8.46 (d, J = 9.2 Hz, 1H), 7.38(td, J = 9.5, 4.6 Hz, 1H), 7.13 (td, J = 9.0, 3.3 Hz, 1H), 3.69 – 3.58 (m,2H), 3.44 – 3.34 (m, 1H), 3.22 (t, J = 7.1 Hz, 1H), 3.12 (t, J = 7.0 Hz, 1H),2.69 – 2.53 (m, 2H), 2.51 – 2.45 (m, 1H), 2.43 (s, 3H), 2.37 – 2.31 (m, 1H),2.30 – 2.20 (m, 1H), 2.01 (dd, J = 13.9, 4.2 Hz, 1H), 1.26 – 1.15 (m, 1H), 0.55 – 0.48 (m, 1H), 0.48 – 0.39 (m, 1H).

[0194] Step Fourteen: N -(5,8-difluoroquinolin-2-yl)-2-(1-methylacetidin-3-yl)-2-azabicyclo[4.1.0]heptane-4-carboxamide (I-23 P2-1 and P2-2)

[0195] Steps eleven to fourteen were performed under the conditions of steps seven to ten in Example 7, using tert-butyl-4-((5,8-difluoroquinolin-2-yl)carbamoyl)-2-azabicyclo[4.1.0]heptane-2-carboxylic acid ester P2: 65 mg (LC-MS retention time 1.39 min) as the starting material to obtainN Chiral resolution of 30 mg of compound I-23 was performed using (5,8-difluoroquinolin-2-yl)-2-(1-methylacetidin-3-yl)-2-azabicyclo[4.1.0]heptane-4-carboxamide) (column: chiralpak-IC, mobile phase: CO2-MeOH(DEA, 0.2%), 40 mL / min, 100 bar) to give fraction P2-1 (5.62 mg) (chiral analysis retention time: 2.38 min) and fraction P2-2 (11.9 mg) (chiral analysis retention time: 2.96 min). Chiral analysis conditions (chiral column: CHIRALPAK OJ-H, 250 mm × 4.6 mm, 5 µm; Methanol (NH4OH) in CO2).

[0196] Component P2-1 analysis data of compound I-23

[0197] LCMS calc. for C 20 H 23 F2N4O [M+H] + m / z = 373.2; Found: 373.1; 1 H NMR (400 MHz, CD3OD) d 8.48 (q, J = 9.2 Hz, 2H), 7.41-7.35 (m, 1H), 7.16-7.11 (m,1H), 3.64 (q, J = 6.4 Hz, 2H), 3.44-3.34 (m, 1H), 3.22 (t, J = 7.1 Hz, 1H), 3.12 (t, J = 7.0 Hz, 1H), 2.69-2.53 (m, 2H), 2.51-2.45 (m, 1H), 2.43 (s, 3H), 2.37-2.31 (m, 1H), 2.29-2.21 (m, 1H), 2.03-1.98 (m, 1H), 1.26-1.15 (m, 1H),0.54-0.42 (m, 2H).

[0198] Component P2-2 analysis data of compound I-23

[0199] LCMS calc. for C 20 H 23 F2N4O [M+H]+ m / z = 373.2; Found: 373.1. 1 H NMR (400 MHz, CD3OD) d 8.45 (s, 2H), 7.38 (td, J = 9.5, 4.6 Hz, 1H), 7.13 (td, J = 9.0, 3.5 Hz, 1H), 3.62 (dt, J = 13.8, 6.0 Hz, 2H), 3.28 – 3.27 (m, 1H), 3.15-3.08 (m, 2H), 2.76-2.70 (m, 1H), 2.65 (dd, J = 11.3, 2.9 Hz, 1H), 2.48 –2.35 (m, 4H), 2.26 – 2.12 (m, 2H), 1.72 – 1.62 (m, 1H), 1.20 – 1.08 (m, 1H), 0.51 – 0.40 (m, 2H).

[0200] Example 7: Preparation of compound I-24

[0201] Note: "or 1" and "or 2" in the structural formula indicate that the compound has a single stereoconfiguration, but its configuration is uncertain.

[0202] Step 1: 1-(tert-butyl)-3-methyl-5-oxopyrrolidine-1,3-dicarboxylic acid ester

[0203] Methyl 5-oxopyrrolidine-3-carboxylate (10.0 g, 69.9 mmol) and DMAP (4.16 g, 34.9 mmol) were dissolved in acetonitrile (100 mL), and (Boc)₂O (22.9 g, 105 mmol) was slowly added. The reaction mixture was stirred at 20 °C for 12 hours, concentrated under reduced pressure to remove acetonitrile, and water (20 mL) was added. The pH was adjusted to 3 with 1 M NaHSO₄ (aq.). The mixture was extracted with ethyl acetate (50 mL x 3). The combined organic phases were dried over anhydrous sodium sulfate and concentrated to give a yellow solid, 1-(tert-butyl)-3-methyl-5-oxopyrrolidine-1,3-dicarboxylate (13.0 g). LCMS calc. for C 11 H 18 NO5 [M+H] +:m / z = 244.1; Found: 244.1.

[0204] Step 2: 1-(tert-butyl)-3-methyl-5-hydroxypyrrolidine-1,3-dicarboxylate

[0205] 1-(tert-butyl)-3-methyl-5-oxopyrrolidine-1,3-dicarboxylic acid ester (13.0 g, 10.1 mmol) was dissolved in tetrahydrofuran (130 mL) and cooled to -80°C. o C, slowly add DIBAL-H (66.8 mL, 66.8 mmol 1 min hexane), and incubate the reaction solution at -80°C. o Stir at C for 1 hour. Add saturated ammonium chloride (80 mL) dropwise to quench the reaction (controlling the temperature <10°C). o C), after stirring for 15 minutes, the mixture was rapidly filtered, separated, and the organic phase was concentrated. Water (50 mL) was added, and the mixture was extracted with diethyl ether (80 mL x 2). The combined organic phases were dried over anhydrous sodium sulfate and concentrated to give a yellow oily crude product, 1-(tert-butyl)-3-methyl-5-hydroxypyrrolidine-1,3-dicarboxylate (8.30 g). LCMS calc. for C 11 H 20 NO5 [M+H] + m / z = 246.1; Found: 246.1.

[0206] Step 3: 1-(tert-butyl)-3-methyl-2,3-dihydro-1 H -Pyrrole-1,3-dicarboxylic acid ester

[0207] 1-(tert-butyl)-3-methyl-5-hydroxypyrrolidine-1,3-dicarboxylate (8.30 g, 6.03 mmol) was dissolved in anhydrous toluene (83 mL) and cooled to -80°C. o C, slowly add TFAA (7.14 g, 33.8 mmol), and after the addition is complete, stop at -80°C. o Stir at C for 20 minutes, then slowly add DIEA (16.6 mL, 128 mmol), and incubate the reaction solution at -80°C. o After naturally heating to room temperature, stir for 12 hours. o Quenching was performed at C with 12 mL of water, pH adjusted to 6 with 10% citric acid (aq.), and extracted with ethyl acetate (20 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated. Purification was achieved by silica gel chromatography (ethyl acetate / petroleum ether = 1:8) to give 1-(tert-butyl)-3-methyl-2,3-dihydro-1-ethylhexylene.H -Pyrrole-1,3-dicarboxylic acid ester (2.45 g). LCMS calc. for C 11 H 18 NO4 [M+H] + : m / z = 228.1; Found: 228.1.

[0208] Step 4: 2-(tert-butyl)-4-methyl-2-azabicyclo[3.1.0]hexane-2,4-dicarboxylic acid ester

[0209] 1-(tert-butyl)3-methyl-2,3-dihydro-1 H 1,3-pyrrole-1,3-dicarboxylic acid ester (2.45 g, 6.03 mmol) was dissolved in anhydrous dichloromethane (60 mL) and heated at -40°C. o C. Add diethylzinc (26.9 mL, 26.9 mmol, 1 M in Hexane) to the reaction solution. After the addition is complete, stir for 5 minutes, then cool the reaction solution to -55°C. o C, add chloroiodomethane (4.9 g, 27.7 mmol), and incubate the reaction solution at -55°C. o C. Allow to heat naturally to room temperature and stir for 12 hours. o At C, a saturated sodium bicarbonate aqueous solution (200 mL) was added to the reaction solution to quench the reaction, followed by filtration, extraction with dichloromethane (20 mL × 3), and the combined organic phases were dried over anhydrous sodium sulfate and concentrated to obtain crude 2-(tert-butyl)-4-methyl-2-azabicyclo[3.1.0]hexane-2,4-dicarboxylic acid ester (1.55 g). LCMS calc. for C 12 H 20 NO4 [M+H] + : m / z = 242.1; Found: 242.1.

[0210] Step 5: 2-(tert-butoxycarbonyl)-2-azabicyclo[3.1.0]hexane-4-carboxylic acid

[0211] 2-(tert-butyl)-4-methyl-2-azabicyclo[3.1.0]hexane-2,4-dicarboxylic acid ester (1.55 g, 1.34 mmol) was dissolved in methanol (65 mL), followed by the addition of sodium hydroxide (0.69 g, 17.4 mmol) and water (6.5 mL). The solution was then heated at 20 °C. oThe mixture was stirred at C for 48 hours. The reaction solution was evaporated to dryness, 40 mL of water was added, and unreacted starting material was extracted with diethyl ether (20 mL x 3). The aqueous phase was adjusted to pH 2 with 1 M NaHSO4 aqueous solution, followed by extraction with chloroform (30 mL x 3). The organic phase was dried over anhydrous sodium sulfate and concentrated to give crude 2-(tert-butoxycarbonyl)-2-azabicyclo[3.1.0]hexane-4-carboxylic acid (0.88 g). LCMS calc. for C 11 H 18 NO4 [M+H] + : m / z = 228.1; Found: 228.1.

[0212] Step 6: tert-butyl 4-((5,8-difluoroquinolin-2-yl)carbamoyl)-2-azabicyclo[3.1.0]hexane-2-carboxylate

[0213] 5,8-Difluoroquinoline-2-amine (840 mg, 4.66 mmol), 1-butylphosphine anhydride (8.90 g, 50% ethyl acetate solution, 14 mmol), and 2-(tert-butoxycarbonyl)-2-azabicyclo[3.1.0]hexane-4-carboxylic acid (1.60 g, 6.99 mmol) were dissolved in pyridine (4 mL), and the reaction solution was heated to 20 °C. o Stirred at C for 3 hours. Add water (20 mL) and extract with ethyl acetate (20 mL x 3). Combine the organic phases, dry and concentrate with anhydrous sodium sulfate, and purify by silica gel chromatography (petroleum ether / ethyl acetate = 3:1) to give two isomers, tert-butyl-4-((5,8-difluoroquinolin-2-yl)carbamoyl)-2-azabicyclo[3.1.0]hexane-2-carboxylate (TLC: less polar fraction P1 (385 mg), more polar fraction P2 (155 mg)). LCMS calc. for C 20 H 22 F2N3O3 [M+H + : m / z = 390.2; Found: 390.2.

[0214] Step Seven: N -(5,8-difluoroquinolin-2-yl)-2-azabicyclo[3.1.0]hexane-4-carboxamide

[0215] 385 mg (0.99 mmol) of tert-butyl-4-((5,8-difluoroquinolin-2-yl)carbamoyl)-2-azabicyclo[3.1.0]hexane-2-carboxylate (the less polar component P1) was dissolved in dichloromethane (3.5 mL), and trifluoroacetic acid (0.7 mL) was added. The reaction solution was heated to 20 °C. o Stir at C for 1 hour. Concentrate the reaction solution to obtain the crude product. N -(5,8-difluoroquinolin-2-yl)-2-azabicyclo[3.1.0]hexane-4-carboxamide (300 mg, TFA salt). LCMS calc. for C 15 H 14 F2N3O[M+H] + : m / z = 290.1; Found: 290.1.

[0216] Step 8: tert-butyl 3-(4-((5,8-difluoroquinolin-2-yl)carbamoyl)-2-azabicyclo[3.1.0]hexane-2-yl)acetidine-1-carboxylic acid ester

[0217] Will N -(5,8-difluoroquinolin-2-yl)-2-azabicyclo[3.1.0]hexane-4-carboxamide (TFA salt) (300 mg, 1.04 mmol) and tert-butyl-3-carbonylacetidine-1-carboxylic acid ester (214 mg, 1.25 mmol) were dissolved in dichloromethane (5 mL), stirred for 20 minutes, and then... o Sodium triacetoxyborohydride (484 mg, 2.28 mmol) was added at C. The reaction was carried out at 20°C. o The mixture was stirred at C for 12 hours. The reaction solution was poured into a saturated sodium bicarbonate aqueous solution (50 mL) and extracted with dichloromethane (20 mL x 3). The combined organic phases were dried over anhydrous sodium sulfate and concentrated, then purified by silica gel chromatography (petroleum ether / ethyl acetate = 3:1) to give a white solid tert-butyl-3-(4-((5,8-difluoroquinolin-2-yl)carbamoyl)-2-azabicyclo[3.1.0]hexane-2-yl)acetidine-1-carboxylic acid ester (145 mg). LCMS calc. for C 23 H 27 F2N4O3 [M+H + : m / z = 445.2; Found: 445.2.

[0218] Step Nine: 2-(Acetidin-3-yl)- N-(5,8-difluoroquinolin-2-yl)-2-azabicyclo[3.1.0]hexane-4-carboxamide

[0219] 145 mg (0.33 mmol) of tert-butyl-3-(4-((5,8-difluoroquinolin-2-yl)carbamoyl)-2-azabicyclo[3.1.0]hexan-2-yl)acetidine-1-carboxylic acid ester was dissolved in dichloromethane (2 mL) and trifluoroacetic acid (0.4 mL), and the reaction was carried out at 20 °C. o The mixture was stirred at C for 1 hour. The reaction solution was concentrated to obtain a crude, colorless oily substance, 2-(acetidin-3-yl)- N -(5,8-difluoroquinolin-2-yl)-2-azabicyclo[3.1.0]hexane-4-carboxamide (145 mg, TFA salt). LCMScalc. for C 18 H 19 F2N4O [M+H] + : m / z = 345.1; Found: 345.1.

[0220] Step 10: N -(5,8-difluoroquinolin-2-yl)-2-(1-methylacetidin-3-yl)-2-azabicyclo[3.1.0]hexane-4-carboxamide

[0221] 2-(acetidine-3-yl)- N -(5,8-difluoroquinolin-2-yl)-2-azabicyclo[3.1.0]hexane-4-carboxamide (145 mg, 0.42 mmol), 30% wt formaldehyde aqueous solution (104 mg, 1.26 mmol) and acetic acid (50.6 mg, 0.84 mmol) were dissolved in methanol (3 mL) at 20 °C. o Sodium cyanoborohydride (79.4 mg, 1.26 mmol) was added at C. The reaction solution was heated to 20 °C. o After stirring at C for 1 hour, 10 mL of saturated sodium bicarbonate aqueous solution was added, and the mixture was extracted with ethyl acetate (20 mL x 3). The combined organic phases were dried over anhydrous sodium sulfate and concentrated. The resulting white solid was purified by preparative high-performance liquid chromatography (mobile phase: ACN-H2O (0.1% NH4HCO3)) to obtain a preparative high-performance liquid chromatography (HPLC) method. N -(5,8-difluoroquinolin-2-yl)-2-(1-methylacetidin-3-yl)-2-azabicyclo[3.1.0]hexane-4-carboxamide (I-16) (32 mg). LCMS calc. for C 19 H 21 F2N4O[M+H] +m / z = 359.2; Found: 359.2.

[0222] Will N -(5,8-difluoroquinolin-2-yl)-2-(1-methylacetidin-3-yl)-2-azabicyclo[3.1.0]hexane-4-carboxamide (32 mg) was chirally resolved (column: CHIRALPAK-IC, mobile phase: CO2-MeOH(DEA, 0.2%), 40 mL / min, 100 bar) to give fraction P1-1 (10.0 mg) of compound I-24 (chiral analysis retention time: 1.42 min) and fraction P1-2 (13.2 mg) of compound I-24 (chiral analysis retention time: 2.02 min). Chiral analysis conditions (chiral column: ChiralCel IC, 500 × 4.6 mm ID, 2 µm; Methanol (NH4OH) in CO2).

[0223] Component P1-1 analysis data of compound I-24

[0224] LCMS calc. for C 19 H 21 F2N4O [M+H] + m / z = 359.2; Found: 359.2. 1 H NMR (400MHz, CD3OD) d 8.51–8.49 (m, 2H), 7.38 (ddd, J = 10.4, 8.7, 4.6 Hz, 1H), 7.14(td, J = 9.0, 3.5 Hz, 1H), 3.70-3.60 (m, 2H), 3.44-3.38 (m, 1H), 3.37-3.33(m, 1H), 3.21 (t, J = 7.3 Hz, 1H), 3.10 (t, J = 7.3 Hz, 1H), 3.01 (dd, J =10.3, 8.1 Hz, 1H), 2.83-2.79 (m, 1H), 2.47-2.39 (m, 4H), 1.97-1.91 (m, 1H), 1.14-1.06 (m, 1H), 0.36-0.31 (m, 1H).

[0225] Analytical data of component P1-2 of compound I-24

[0226] LCMS calc. for C 19 H 21 F2N4O [M+H] + m / z = 359.2; Found: 359.2. 1 H NMR (400MHz, CD3OD) d 8.55-8.43 (m, 2H), 7.38 (ddd, J = 10.3, 8.7, 4.6 Hz, 1H), 7.13(td, J = 9.0, 3.4 Hz, 1H), 3.73-3.62 (m, 2H), 3.45-3.37 (m, 1H), 3.38 -3.33(m, 1H), 3.26 (t, J = 7.2 Hz, 1H), 3.15 (t, J = 7.3 Hz, 1H), 3.01 (dd, J =10.2, 8.1 Hz, 1H), 2.84-2.80 (m, 1H), 2.45 (s, 3H), 2.42 (t, J = 10.4 Hz,1H), , 1.97-1.91 (m, 1H), 1.11-1.08 (m, 1H), 0.35-0.30 (m, 1H).

[0227] Biological experiments

[0228] cGMP time-resolved fluorescence assay: When ANP or BNP binds to NPR1, it activates its guanylate cyclase, converting GTP to cGMP. Therefore, the activity of NPR1 can be characterized by detecting the generation of intracellular cGMP using a time-resolved fluorescence kit (Revvity, 62GM2PEG).

[0229] Cell line construction and maintenance

[0230] HEK293 cells were purchased from Nanjing Kebai Biotechnology Co., Ltd., and were correctly identified by short tandem repeat (STR) fragments. NPR1The gene was synthesized by Ginkgo Biologics. After packaging the virus, HEK293 cells were infected according to standard viral infection procedures, followed by antibiotic screening to obtain stable NPR1 overexpression transgenic cells. The stable transgenic cells were cultured in DMEM (Gibco, C11995500BT) medium supplemented with 10% fetal bovine serum (Gibco, 10099141C), 1% penicillin-streptomycin (Gibco, 15140-122), and 2 μg / mL puromycin (Beyotime, ST551).

[0231] Compound testing

[0232] Prepare experimental buffer containing 44 pM ANP (Targetmol, TP1220L): Opti-MEM (Gibco, 11058-021) + 2% fetal bovine serum + 1 mM IBMX (Targetmol, T1713). Seed 5 μL of cell suspension containing 5000 NPR1-overexpressing HEK293 stable transgenic cells into 384-well flat-bottomed plates (PE, ProxiPlate-384 plus) pre-filled with 5 μL of experimental buffer per well. Add compounds in an incremental concentration gradient, with 5 nM ANP treatment as a control. Centrifuge at 1000 rpm for 1 minute. After incubating at room temperature for 30 minutes, add 5 μL of d2-labeled cGMP and 5 μL of Eu-labeled cGMP antibody. After incubating at room temperature in the dark for 60 minutes, measure the time-resolved fluorescence signals at 620 nm and 665 nm using a Spark (Tecan) microplate reader. 5 nM ANP is the minimum value (Min well), with relative activity set to 100%; DMSO treatment is the maximum value (Max well), with relative activity set to 0%. Calculation formula used:

[0233] The relative activity of each group was expressed as a percentage change relative to the DMSO control group, and then fitted to a four-parameter nonlinear curve using the program GraphPadprism 10.

[0234] Table 1 + refers to IC 50 ≤50 nM, ++ means 50 nM < IC 50 ≤500 nM, +++ means 500 nM <IC 50 ≤5000 nM, ++++ refers to IC 50 >5000 nM.

[0235] The structure of Ref-1 is Selected from WO2024233501 A2.

Claims

1. A compound of formula (I), its isotopic label, enantiomer, diastereomer, transisomer, or pharmaceutically acceptable salt thereof: in, It can be a single bond or a double bond; X 1 For N, NR x1 CR 1 or CR 1 R 1a X 2 For N, NR x2 CR 2 or CR 2 R 2a X 3 For N or CR 3 X 4 For N or CR 4 X 5 For N or CR 5 X 6 For N or CR 6 ; R x1 and R x2 Each is independently selected from hydrogen, C1-C6 alkyl, halo-C1-C6 alkyl, and deuterated C1-C6 alkyl; R 1 R 1a R 2 R 2a R 3 R 4 R 5 R 6 Each is independently selected from hydrogen, deuterium, halogen, cyano, carboxyl, hydroxyl, -NR x R y -C(=O)NR x R y -NR x C(=O)R y -C(=O)OR z -C(=O)R z -S(=O)2R z -C(=O)NH-S(=O)2R z -P(=O) R z R t No substitution or by one or more R A Substituted C1-C6 alkyl, unsubstituted, or with one or more R A Substituted C1-C6 alkoxy groups, unsubstituted, or substituted with one or more R groups A Substituted C2-C6 alkenyl groups, unsubstituted, or with one or more R groups A Substituted C2-C6 ynyl group, unsubstituted, or with one or more R groups A Substituted C3-C6 cycloalkyl, unsubstituted, or with one or more R A Substituted C6-C8 aryl groups, unsubstituted or with one or more R groups A Substituted 4-6 membered heterocyclic groups, unsubstituted or with one or more R groups A Substituted 5-6 aryl groups; R A Selected from C1-C6 alkyl, halogenated C1-C6 alkyl, deuterated C1-C6 alkyl, C1-C6 alkoxy, halogenated C1-C6 alkoxy, deuterated C1-C6 alkoxy, deuterium, halogen, carboxyl, hydroxyl, cyano, -NR A1 R A2 -C(=O)NR A1 R A2 -C(=O)OR A1 -NR A1 C(=O)R A2 ; L 1 Selected from , or ;in, Indicates with L 2 The connected side; R L1 It is hydrogen or C1-C6 alkyl; L 2 -(CR) 7 R 8 ) i -; Each R 7 and R 8 Each of these can be independently hydrogen, deuterium, halogen, cyano, carboxyl, hydroxyl, C1-C6 alkyl, halo-C1-C6 alkyl, or deuterated C1-C6 alkyl; i can be 0, 1, 2, or 3. W is selected from unsubstituted or R-selected. B Substituted C6-C10 cycloalkyl, unsubstituted, or with one or more R B Substituted 5-10 membered heterocyclic groups; the 5-10 membered heterocyclic group contains 1-2 N atoms; R B Selected from oxo (=O), deuterium, halogen, hydroxyl, cyano, -NR B1 R B2 -C(=O)NR B1 R B2 -C(=O)OR B1 -NR B1 C(=O)R B2 、-(CR 9 R 10 ) n -R 11 -OR 12 ; n is 0, 1, 2, or 3; each R 9 and R 10 Each can be independently hydrogen, deuterium, halogen, cyano, carboxyl, hydroxyl, C1-C6 alkyl, halo-C1-C6 alkyl, or deuterated C1-C6 alkyl. R 11 and R 12 Selected from unsubstituted or by one or more R C Substituted C1-C6 alkyl, unsubstituted, or with one or more R C Substituted C2-C6 alkenyl groups, unsubstituted, or with one or more R groups C Substituted C2-C6 ynyl group, unsubstituted, or with one or more R groups C Substituted C3-C6 cycloalkyl, unsubstituted, or with one or more R C Substituted C6-C10 aryl groups, unsubstituted, or substituted with one or more R groups C Substituted 4-6 membered heterocyclic groups, unsubstituted or with one or more R groups C Substituted 5-6 aryl groups; R C Selected from oxo (=O), deuterium, halogen, hydroxyl, cyano, C1-C6 alkyl, deuterated C1-C6 alkyl, halo-C1-C6 alkyl, hydroxyl-substituted C1-C6 alkyl, cyano-substituted C1-C6 alkyl, C1-C6 alkoxy-substituted C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, halo-C1-C6 alkoxy, C3-C6 cycloalkyl, C3-C6 cycloalkoxy, -NR C1 R C2 -C(=O)NR C1 R C2 -C(=O)OR C1 -NR C1 C(=O)R C2 -SO2R C1 ; Or two adjacent R B Together with the atoms between them, they form unsubstituted or substituted groups with one or more R atoms. D Substituted phenyl, unsubstituted, or substituted with one or more R D Substituted 5-6 aryl groups; R D Selected from C1-C6 alkyl, halogenated C1-C6 alkyl, deuterated C1-C6 alkyl, C1-C6 alkoxy, halogenated C1-C6 alkoxy, deuterated C1-C6 alkoxy, deuterium, halogen, hydroxyl, carboxyl, cyano, -NR D1 R D2 -C(=O)NR D1 R D2 -NR D1 C(=O)R D2 ; R x R y R A1 R A2 R B1 R B2 R C1 R C2 R D1 R D2 Each is independently selected from hydrogen, C3-C6 cycloalkyl, unsubstituted, or by one or more R E Substituted C1-C6 alkyl groups; R E Selected from deuterium, halogens, C1-C6 alkoxy groups, deuterated C1-C6 alkoxy groups, halogenated C1-C6 alkoxy groups, and -NR. E1 R E2 ;R E1 R E2 Each is independently selected from hydrogen, C1-C6 alkyl, and C3-C6 cycloalkyl; R z R t Each is independently selected from C1-C6 alkyl groups; The 4-6 membered heterocyclic group contains 1-3 heteroatoms selected from N, O, and S, and the 5-6 membered heteroaryl group contains 1-3 heteroatoms selected from N, O, and S.

2. The compound of formula (I) as claimed in claim 1, its isotopic label, enantiomer, diastereomer, transisomer, or pharmaceutically acceptable salt thereof, characterized in that, The compound shown in formula (I) has the following structure: Among them, X 2 X 4 R 1 R 3 R 5 R 6 L 1 L 2 The definition of W is the same as that in claim 1.

3. The compound of formula (I) as claimed in claim 1, its isotopic label, enantiomer, diastereomer, transisomer, or pharmaceutically acceptable salt thereof, characterized in that, The compound shown in formula (I) has the following structure: Among them, R 1 R 2 R 3 R 4 R 5 R 6 L 1 L 2 The definition of W is the same as that in claim 1.

4. The compound of formula (I) as claimed in any one of claims 1-3, its isotopic label, enantiomer, diastereomer, transisomer, or pharmaceutically acceptable salt thereof, characterized in that, R 1 R 2 R 3 R 4 R 5 R 6 Each is independently selected from hydrogen, deuterium, halogen, cyano, carboxyl, hydroxyl, amino, C1-C6 alkyl, C1-C6 alkoxy, halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, deuterated C1-C6 alkyl, and deuterated C1-C6 alkoxy.

5. The compound of formula (I) according to any one of claims 1-3, its isotopic label, enantiomer, diastereomer, transisomer, or pharmaceutically acceptable salt thereof, characterized in that, Selected from: 、 、 、 、 、 、 、 、 。 6. The compound of formula (I) as claimed in any one of claims 1-3, its isotopic label, enantiomer, diastereomer, transisomer, or pharmaceutically acceptable salt thereof, characterized in that, L 1 for or L 2 For a bond or -CH2-; preferably, L 1 for L 2 As the key; where, Indicates with L 2 The connected side.

7. The compound, isotope label thereof, enantiomer, diastereomer, transisomer, or pharmaceutically acceptable salt thereof, as shown in (I) of any one of claims 1-3, characterized in that, W is selected from unsubstituted or R-selected. B The following groups are substituted: 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 ； R B Selected from deuterium, halogen, hydroxyl, cyano, -NR B1 R B2 -C(=O)NR B1 R B2 -C(=O)OR B1 -NR B1 C(=O)R B2 、-(CR 9 R 10 ) n -R 11 -OR 12 ; n is 0, 1, 2, or 3; each R 9 and R 10 Each can be independently hydrogen, deuterium, halogen, cyano, carboxyl, hydroxyl, C1-C6 alkyl, halo-C1-C6 alkyl, or deuterated C1-C6 alkyl. R 11 and R 12 Selected from unsubstituted or by one or more R C Substituted C1-C6 alkyl, unsubstituted, or with one or more R C Substituted C2-C6 alkenyl groups, unsubstituted, or with one or more R groups C Substituted C2-C6 ynyl group, unsubstituted, or with one or more R groups C Substituted C3-C6 cycloalkyl, unsubstituted, or with one or more R C Substituted C6-C10 aryl groups, unsubstituted, or substituted with one or more R groups C Substituted 4-6 membered heterocyclic groups, unsubstituted or with one or more R groups C Substituted 5-6 aryl groups; R C Selected from oxo (=O), deuterium, halogen, hydroxyl, cyano, C1-C6 alkyl, deuterated C1-C6 alkyl, halo-C1-C6 alkyl, hydroxyl-substituted C1-C6 alkyl, cyano-substituted C1-C6 alkyl, C1-C6 alkoxy-substituted C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, halo-C1-C6 alkoxy, C3-C6 cycloalkyl, C3-C6 cycloalkoxy, -NR C1 R C2 -C(=O)NR C1 R C2 -C(=O)OR C1 -NR C1 C(=O)R C2 -SO2R C1 ; R B1 R B2 R C1 R C2 Each is independently selected from hydrogen, C3-C6 cycloalkyl, unsubstituted, or by one or more R E Substituted C1-C6 alkyl groups; R E Selected from deuterium, halogens, C1-C6 alkoxy groups, deuterated C1-C6 alkoxy groups, halogenated C1-C6 alkoxy groups, and -NR. E1 R E2 ;R E1 R E2 Each is independently selected from hydrogen, C1-C6 alkyl, and C3-C6 cycloalkyl; The 4-6 membered heterocyclic group contains 1-3 heteroatoms selected from N, O, and S, and the 5-6 membered heteroaryl group contains 1-3 heteroatoms selected from N, O, and S.

8. The compound of formula (I) as claimed in any one of claims 1-3, its isotopic label, enantiomer, diastereomer, transisomer, or pharmaceutically acceptable salt thereof, characterized in that, W is selected from: 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 ； m can be 0, 1, 2, 3, or 4; R 13 Selected from deuterium, halogen, hydroxyl, cyano, -NR 13A R 13B -C(=O)NR 13A R 13B -C(=O)OR 13A -NR 13A C(=O)R 13B 、-(CR 15 R 16 ) p -R 17 -OR 18 ; p is 0, 1, 2, or 3; each R 15 and R 16 Each can be independently hydrogen, deuterium, halogen, cyano, carboxyl, hydroxyl, C1-C6 alkyl, halo-C1-C6 alkyl, or deuterated C1-C6 alkyl. R 17 and R 18 Selected from unsubstituted or by one or more R 19 Substituted C1-C6 alkyl, unsubstituted, or with one or more R 19 Substituted C2-C6 alkenyl groups, unsubstituted, or with one or more R groups 19 Substituted C2-C6 ynyl group, unsubstituted, or with one or more R groups 19 Substituted C3-C6 cycloalkyl, unsubstituted, or with one or more R 19 Substituted C6-C10 aryl groups, unsubstituted, or substituted with one or more R groups 19 Substituted 4-6 membered heterocyclic groups, unsubstituted or with one or more R groups 19 Substituted 5-6 aryl groups; R 19 Selected from oxo (=O), deuterium, halogen, hydroxyl, cyano, C1-C6 alkyl, deuterated C1-C6 alkyl, halo-C1-C6 alkyl, hydroxyl-substituted C1-C6 alkyl, cyano-substituted C1-C6 alkyl, C1-C6 alkoxy-substituted C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, halo-C1-C6 alkoxy, C3-C6 cycloalkyl, C3-C6 cycloalkoxy, -NR 19A R 19B -C(=O)NR 19A R 19B -C(=O)OR 19A -NR 19A C(=O)R 19B ; R 14 For hydrogen or -(CR) 20 R 21 ) q -R 22 ; q is 0, 1, 2, or 3; each R 20 and R 21 Each can be independently hydrogen, deuterium, halogen, cyano, carboxyl, hydroxyl, C1-C6 alkyl, halo-C1-C6 alkyl, or deuterated C1-C6 alkyl. R 22 Selected from unsubstituted or by one or more R 23 Substituted C1-C6 alkyl, unsubstituted, or with one or more R 23 Substituted C3-C6 cycloalkyl, unsubstituted, or with one or more R 23 Substituted 4-6 membered heterocyclic groups, unsubstituted or with one or more R groups 23 Substituted 5-6 aryl groups; R 23 Selected from oxo (=O), deuterium, halogen, hydroxyl, cyano, C1-C6 alkyl, deuterated C1-C6 alkyl, halo-C1-C6 alkyl, hydroxyl-substituted C1-C6 alkyl, cyano-substituted C1-C6 alkyl, C1-C6 alkoxy-substituted C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, halo-C1-C6 alkoxy, C3-C6 cycloalkyl, C3-C6 cycloalkoxy, -NR 23A R 23B -C(=O)NR 23A R 23B -C(=O)OR 23A -NR 23A C(=O)R 23B -SO2R 23A ; R 13A R 13B R 19A R 19B R 23A R 23B Each is independently selected from hydrogen, C3-C6 cycloalkyl, unsubstituted, or by one or more R F Substituted C1-C6 alkyl groups; R F Selected from deuterium, halogens, C1-C6 alkoxy groups, deuterated C1-C6 alkoxy groups, halogenated C1-C6 alkoxy groups, and -NR. F1 R F2 ;R F1 R F2 Each is independently selected from hydrogen, C1-C6 alkyl, and C3-C6 cycloalkyl; The 4-6 membered heterocyclic group contains 1-3 heteroatoms selected from N, O, and S, and the 5-6 membered heteroaryl group contains 1-3 heteroatoms selected from N, O, and S.

9. The compound of formula (I) as claimed in any one of claims 1-3, its isotopic label, enantiomer, diastereomer, transisomer, or pharmaceutically acceptable salt thereof, characterized in that, W is selected from: 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 ； m can be 0, 1, 2, 3, or 4; R 13 Selected from halogen, hydroxyl, cyano, -NR 13A R 13B -C(=O)NR 13A R 13B -C(=O)OR 13A -NR 13A C(=O)R 13B 、-(CR 15 R 16 ) p -R 17 -OR 18 ; p is 0, 1, 2, or 3; each R 15 and R 16 Each is independently hydrogen or C1-C6 alkyl; R 17 and R 18 Selected from unsubstituted or by one or more R 19 Substituted C1-C6 alkyl, unsubstituted, or with one or more R 19 Substituted C2-C6 alkenyl groups, unsubstituted, or with one or more R groups 19 Substituted C2-C6 ynyl group, unsubstituted, or with one or more R groups 19 Substituted C3-C6 cycloalkyl, unsubstituted, or with one or more R 19 Substituted C6-C10 aryl groups, unsubstituted, or substituted with one or more R groups 19 Substituted 4-6 membered heterocyclic groups, unsubstituted or with one or more R groups 19 Substituted 5-6 aryl groups; R 19 Selected from oxo (=O), deuterium, halogen, hydroxyl, cyano, C1-C6 alkyl, deuterated C1-C6 alkyl, halo-C1-C6 alkyl, hydroxyl-substituted C1-C6 alkyl, cyano-substituted C1-C6 alkyl, C1-C6 alkoxy-substituted C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, halo-C1-C6 alkoxy, C3-C6 cycloalkyl, C3-C6 cycloalkoxy, -NR 19A R 19B -C(=O)NR 19A R 19B -C(=O)OR 19A -NR 19A C(=O) R 19B ; R 13A R 13B R 19A R 19B Each is independently selected from hydrogen, C3-C6 cycloalkyl, unsubstituted, or by one or more R F Substituted C1-C6 alkyl groups; R F Selected from deuterium, halogens, C1-C6 alkoxy groups, deuterated C1-C6 alkoxy groups, halogenated C1-C6 alkoxy groups, and -NR. F1 R F2 ;R F1 R F2 Each is independently selected from hydrogen, C1-C6 alkyl, and C3-C6 cycloalkyl; The 4-6 membered heterocyclic group contains 1-3 heteroatoms selected from N, O, and S, and the 5-6 membered heteroaryl group contains 1-3 heteroatoms selected from N, O, and S; R 14 Selected from hydrogen, -CH2N(CH3)2, -CH2CH2N(CH3)2, -CH2CH2CH2N(CH3)2, , , , , , , , , , , , , , , , -SO2CH3.

10. The compound of formula (I) as claimed in any one of claims 1-3, its isotopic label, enantiomer, diastereomer, transisomer, or pharmaceutically acceptable salt thereof, characterized in that, W is selected from: , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , .

11. The compound of formula (I) as claimed in claim 1, its isotopic label, enantiomer, diastereomer, transisomer, or pharmaceutically acceptable salt thereof, characterized in that, The compound represented by formula (I) is selected from the following structures: in, s is 0, 1, or 2; t is 0, 1, or 2; X 2 For N or CR 2 X 4 For N or CR 4 ; R 1 R 2 R 3 R 4 R 5 R 6 Each is independently selected from hydrogen, deuterium, halogen, cyano, carboxyl, hydroxyl, amino, C1-C6 alkyl, C1-C6 alkoxy, halo-C1-C6 alkyl, halo-C1-C6 alkoxy, deuterated C1-C6 alkyl, and deuterated C1-C6 alkoxy. R 24 For hydrogen or -(CR) 25 R 26 ) y -R 27 ; y is 0, 1, 2, or 3; each R 25 and R 26 Each can be independently hydrogen, deuterium, halogen, cyano, carboxyl, hydroxyl, C1-C6 alkyl, halo-C1-C6 alkyl, or deuterated C1-C6 alkyl. R 27 Selected from unsubstituted or by one or more R 28 Substituted C1-C6 alkyl, unsubstituted, or with one or more R 28 Substituted 4-6 membered heterocyclic groups, unsubstituted or with one or more R groups 28 Substituted 5-6 aryl groups; R 28 Selected from oxo (=O), deuterium, halogen, hydroxyl, cyano, C1-C6 alkyl, deuterated C1-C6 alkyl, halo-C1-C6 alkyl, hydroxyl-substituted C1-C6 alkyl, cyano-substituted C1-C6 alkyl, C1-C6 alkoxy-substituted C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, halo-C1-C6 alkoxy, C3-C6 cycloalkyl, C3-C6 cycloalkoxy, -NR 28A R 28B -C(=O)NR 28A R 28B -C(=O)OR 28A -NR 28A C(=O)R 28B -SO2R 28A ; R 28A R 28B Each is independently selected from hydrogen, C3-C6 cycloalkyl, unsubstituted, or by one or more R G Substituted C1-C6 alkyl groups; R G Selected from deuterium, halogens, C1-C6 alkoxy groups, deuterated C1-C6 alkoxy groups, halogenated C1-C6 alkoxy groups, and -NR. G1 R G2 ;R G1 R G2 Each is independently selected from hydrogen, C1-C6 alkyl, and C3-C6 cycloalkyl; The 4-6 membered heterocyclic group contains 1-3 heteroatoms selected from N, O, and S, and the 5-6 membered heteroaryl group contains 1-3 heteroatoms selected from N, O, and S.

12. The compound of formula (I) as claimed in claim 11, its isotopic label, enantiomer, diastereomer, transisomer, or pharmaceutically acceptable salt thereof, characterized in that, R 24 Selected from hydrogen, -CH2N(CH3)2, -CH2CH2N(CH3)2, -CH2CH2CH2N(CH3)2, , , , , , , , , , , , , , , , -SO2CH3.

13. The compound of formula (I) as claimed in claim 11, its isotopic label, enantiomer, diastereomer, transisomer, or pharmaceutically acceptable salt thereof, characterized in that, The compound represented by formula (I) is selected from the following compounds: 。 14. A pharmaceutical composition comprising a compound of formula (I) as claimed in any one of claims 1-13, an isotopic label thereof, an enantiomer, a diastereomer, a transisomer thereof, or a pharmaceutically acceptable salt thereof, and a pharmaceutical excipient.

15. The use of a compound of formula (I) as described in any one of claims 1-13, an isotopic label thereof, an enantiomer, a diastereomer, a transisomer thereof, or a pharmaceutically acceptable salt thereof, in the preparation of an NPR1 positive allosteric modulator or a medicament; said medicament being a medicament for treating and / or preventing disease by modulating NPR1.

16. The use of a compound of formula (I) as described in any one of claims 1-13, its isotopic label, enantiomer, diastereomer, transisomer or pharmaceutically acceptable salt thereof in the preparation of a medicament for the treatment and / or prevention of cardiovascular diseases.

17. A method for treating and / or preventing cardiovascular disease, characterized in that, The compound, its isotope label, enantiomer, diastereomer, transisomer or pharmaceutically acceptable salt thereof, as described in any one of claims 1-13, is administered to the patient in a therapeutically effective amount.

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