Tricyclic compound as myosin ii inhibitor and use thereof

Abstract

Disclosed in the present invention are a benzo-cyclic compound represented by formula (I), or a stereoisomer or a pharmaceutically acceptable salt and a pharmaceutical composition comprising same, and a use thereof in the preparation of a drug for treating / preventing a Myosin II-mediated disease, wherein groups in formula (I) are as defined in the description.

Description

Tricyclic compounds as Myosin II inhibitors and their uses Technical Field

[0001] The present invention relates to a Myosin II inhibitor, its stereoisomers, pharmaceutically acceptable salts, solvates, cocrystals or deuterated substances, and use thereof in preparing drugs for treating Myosin II-mediated related diseases. Background Art

[0002] Skeletal muscle plays two crucial roles in the human body: first, muscle contraction, which produces movement and maintains posture; and second, skeletal muscle is the site of glucose, fatty acid, and amino acid metabolism. During normal daily activities, skeletal muscle contraction is closely associated with muscle stress, breakdown, and remodeling, all of which are critical for muscle adaptation. However, in patients with progressive muscular dystrophies, such as Duchenne muscular dystrophy (DMD), muscle contraction leads to repeated rounds of irreparable, amplified muscle breakdown. As patients age, these changes eventually accumulate and develop into a pathological process, leading to excessive inflammation, fibrosis, and the accumulation of fat deposits in the muscles, which in turn progresses to a sharp decline in physical function and ultimately death.

[0003] DMD is a genetic disease affecting skeletal muscle. Becker muscular dystrophy (BMD), a variant of DMD, was first described by German physician Peter Emil Becker in the 1950s. Both are characterized by progressive muscle degeneration and weakness. Currently, there is a need for drugs that can treat patients with DMD or BMD. Summary of the Invention

[0004] The present invention provides a compound of formula (I), formula (II), formula (II-1), formula (I-1), and formula (I-2), and a stereoisomer or pharmaceutically acceptable salt thereof. The compound has the excellent effects of good activity, excellent physicochemical properties, ease of formulation, excellent pharmacokinetic properties, high bioavailability, and low toxic and side effects.

[0005] The present invention provides compounds of formula (I), formula (II), formula (II-1), formula (I-1), and formula (I-2), stereoisomers, or pharmaceutically acceptable salts thereof.

[0006] in,

[0007] R is selected from -O-halogenated C 1-4 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, -O-(CH2) r -R a 、C 3-10Cycloalkyl, 5-10 membered heteroaryl containing 1-3 heteroatoms selected from N, O, S, -C 1-4 Alkyl-R a 、-C 1-4 Alkyl-OR a 、-C 1-4 Alkyl-NR b R a 、-NR b -S(O)2-R a 、-NR b -S(O)2-NR b R a 、-O-NR b R a 、-NH-OR b 、-NH-halogenated C 1-4 Alkyl, -OC 1-4 Alkyl, C 1-6 Alkoxy, halogen, cyano, nitro, C 1-4 Alkyl, -NH-C 1-4 Alkyl, -N(C 1-4 Alkyl)2, -NH-C 3-10 Cycloalkyl, -NHC(O)C 1-4 Alkyl, 4-10 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, S, -CO-C 1-4 Alkyl, -CO-R a 、-S(O)-R a 、-S(O)2-R a The alkyl, alkoxy, alkenyl, alkynyl, heteroaryl, cycloalkyl, heterocycloalkyl may be further substituted by 1-3 halogen, D, CN, OH, C 1-4 Alkyl, halogenated C 1-4 Alkyl, deuterated C 1-4 Alkyl, C 1-4 Alkoxy, halogenated C 1-4 Alkoxy, deuterated C 1-4 Alkoxy, C 3-10 Cycloalkyl, 4-6 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, S, phenyl and NH2; in some embodiments, R is selected from -O-halogenated C 1-4 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, -O-(CH2) r -R a 、C 3-10 Cycloalkyl, 5-10 membered heteroaryl containing 1-3 heteroatoms selected from N, O, S, -C 1-4 Alkyl-R a 、-C 1-4 Alkyl-ORa 、-C 1-4 Alkyl-NR b R a 、-NR b -S(O)2-R a 、-NR b -S(O)2-NR b R a 、-O-NR b R a 、-NH-OR b 、-NH-halogenated C 1-4 Alkyl, -OC 1-4 Alkyl, C 1-6 Alkoxy, halogen, cyano, nitro, C 1-4 Alkyl, -NH-C 1-4 Alkyl, -N(C 1-4 Alkyl)2, -NH-C 3-10 Cycloalkyl, -NHC(O)C 1-4 Alkyl or 4-10 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, S, wherein the alkyl, alkoxy, alkenyl, alkynyl, heteroaryl, cycloalkyl, heterocycloalkyl is optionally further substituted by 1-3 heteroatoms selected from halogen, D, CN, OH, C 1-4 Alkyl, halogenated C 1-4 Alkyl, deuterated C 1-4 Alkyl, C 1-4 Alkoxy, halogenated C 1-4 Alkoxy, deuterated C 1-4 Alkoxy, C 3-10 Cycloalkyl, 4-6 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, S, phenyl, and NH2; in some embodiments, R is selected from -O-halogenated C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, -O-(CH2) r -R a 、C 3-8 Cycloalkyl, 5-10 membered heteroaryl containing 1-3 heteroatoms selected from N, O, S, -C 1-2 Alkyl-R a 、-C 1-2 Alkyl-OR a 、-C 1-2 Alkyl-NR b R a 、-NR b -S(O)2-R a 、-NR b -S(O)2-NR b R a 、-O-NR b Ra 、-NH-OR b 、-NH-halogenated C 1-2 Alkyl, -OC 1-2 Alkyl, C 1-4 Alkoxy, halogen, cyano, nitro, C 1-4 Alkyl, -NH-C 1-2 Alkyl, -N(C 1-2 Alkyl)2, -NH-C 3-8 Cycloalkyl, -NHC(O)C 1-2 Alkyl or 4-10 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, S, wherein the alkyl, alkoxy, alkenyl, alkynyl, heteroaryl, cycloalkyl, heterocycloalkyl is optionally further substituted by 1-3 heteroatoms selected from halogen, D, CN, OH, C 1-2 Alkyl, halogenated C 1-2 Alkyl, deuterated C 1-2 Alkyl, C 1-2 Alkoxy, halogenated C 1-2 Alkoxy, deuterated C 1-2 Alkoxy, C 3-6 Cycloalkyl, 4-6 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, S, phenyl, and NH2; in some embodiments, R is selected from -CF3, -CH2CH3, -CH2CH2CH3, -COCH3, -COCF3, In some embodiments, R is selected from -CF3, -CH2CH3, -CH2CH2CH3, In certain embodiments, R is selected from -O-haloC 1-4 Alkyl, wherein the alkyl is optionally further substituted by 1-3 groups selected from halogen, D, CN, OH, C 1-4 Alkyl, halogenated C 1-4 Alkyl, deuterated C 1-4 Alkyl, C 1-4 Alkoxy, halogenated C 1-4 Alkoxy, deuterated C 1-4 Alkoxy and NH2 groups are substituted; in certain embodiments, R is selected from In certain embodiments, R is selected from -COCH3, -COCF3, -COCH3CF3,

[0008] R a Selected from CN, C 3-10 Cycloalkyl, 4-10 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, S, 6-10 membered aryl, 5-10 membered heteroaryl containing 1-3 heteroatoms selected from N, O, S, -C(O)-R a1 The cycloalkyl, heterocycloalkyl, aryl, heteroaryl groups are optionally further substituted by 1-3 groups selected from halogen, =O, deuterium, CN, OH, C 1-4 Alkyl, halogenated C 1-4 Alkyl, deuterated C 1-4 Alkyl, C 1-4 Alkoxy, halogenated C 1-4 Alkoxy, deuterated C 1-4 Alkoxy and NH2 groups are substituted; in some embodiments, in certain embodiments, each R a Selected from CN, halogenated C 1-4 Alkyl, C 3-6 Monocyclic cycloalkyl, C 7-10 Spirocyclic cycloalkyl, C 4-8 Bridged ring cycloalkyl, C 4-9 cycloalkyl, 4-6 membered monocyclic heterocycloalkyl containing 1-3 heteroatoms selected from N, O, and S, 7-10 membered spirocyclic heterocycloalkyl containing 1-3 heteroatoms selected from N, O, and S, 4-8 membered bridged heterocycloalkyl containing 1-3 heteroatoms selected from N, O, and S, 4-6 membered cycloheterocycloalkyl containing 1-3 heteroatoms selected from N, O, and S, phenyl, 8-10 membered aryl, 5-6 membered monocyclic heteroaryl containing 1-3 heteroatoms selected from N, O, and S, 8-10 membered bicyclic heteroaryl containing 1-3 heteroatoms selected from N, O, and S, -C(O)-R a1 The alkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl groups are optionally further substituted by 1-3 groups selected from halogen, =O, deuterium, CN, OH, C 1-2 Alkyl, halogenated C 1-2 Alkyl, deuterated C 1-2 Alkyl, C 1-2 Alkoxy, halogenated C 1-2 Alkoxy, deuterated C 1-2 Alkoxy and NH2 group substitution;

[0009] In certain embodiments, each R a Selected from CN, halogenated C 1-4 Alkyl, C 3-6Monocyclic cycloalkyl, 4-6 membered monocyclic heterocycloalkyl containing 1-3 heteroatoms selected from N, O, and S, 7-10 membered spirocyclic heterocycloalkyl containing 1-3 heteroatoms selected from N, O, and S, 4-8 membered bridged heterocycloalkyl containing 1-3 heteroatoms selected from N, O, and S, 4-6 membered cyclic heterocycloalkyl containing 1-3 heteroatoms selected from N, O, and S, phenyl, 8-10 membered aryl, 5-6 membered monocyclic heteroaryl containing 1-3 heteroatoms selected from N, O, and S, 8-10 membered bicyclic heteroaryl containing 1-3 heteroatoms selected from N, O, and S, -C(O)-R a1 The alkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl groups are optionally further substituted by 1-3 groups selected from halogen, =O, deuterium, CN, OH, C 1-2 Alkyl, halogenated C 1-2 Alkyl, deuterated C 1-2 Alkyl, C 1-2 Alkoxy, halogenated C 1-2 Alkoxy, deuterated C 1-2 Alkoxy and NH2 group substitution;

[0010] In certain embodiments, each R a Selected from CN, -CH2F, -CHF2, -CF3, -CH2CH2F, -CH2CHF2, -CH2CF3, -CHFCH2F, -CHFCHF2, -CHFCF3, -CF2CH2F, -CF2CHF2, -CF2CF3, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, azetidinyl, azetyl, oxetanyl, oxetanyl, piperazinyl, piperidinyl, tetrahydropyranyl, morpholinyl, pyranyl, phenyl, pyrrolyl, furyl, thienyl, imidazolyl, oxazolyl, thiazolyl, pyrazolyl, isoxazolyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, -C(O)-R a1, wherein the above groups are optionally further replaced by 1, 2, or 3 groups selected from F, Cl, =O, deuterium, CN, OH, methyl, ethyl, -CH2F, -CHF2, -CF3, -CH2CH2F, -CH2CHF2, -CH2CF3, -CHFCH2F, -CHFCHF2, -CHFCF3, -CF2CH2F, -CF2CHF2, -CF2CF3, -CH2D, -CHD2, -CD3, -CH2CH2D, -CH2CHD2, -CH2CD3, -CHDCH2D, -CHDCHD2, -CHDCD3, -CD2CH2D, -CD2CHD2, -CD2CD 3. methoxy, ethoxy, -OCHF2, -OCH2F, -OCF3, -OCH2CH2F, -OCH2CHF2, -OCH2CF3, -OCHFCH2F, -OCHFCHF2, -OCHFCF3, -OCF2CH2F, -OCF2CHF2, -OCF2CF3, -OCHD2, -OCH2D, -OCD3, -OCH2CH2D, -OCH2CHD2, -OCH2CD3, -OCHDCH2D, -OCHDCHD2, -OCHDCD3, -OCD2CH2D, -OCD2CHD2, -OCD2CD3 and NH2;

[0011] Each R b Selected from H, deuterium, C 1-4 Alkyl, C 3-6 Cycloalkyl, halogenated C 1-4 Alkyl, deuterated C 1-4 Alkyl; in certain embodiments, each R b Selected from H, deuterium, C 1-4 Alkyl, C 3-6 Cycloalkyl; in certain embodiments, each R b Selected from H, deuterium, C 1-2 Alkyl, C 3-4 Cycloalkyl; in certain embodiments, each R b is selected from H, deuterium, and methyl;

[0012] R a1 Selected from OH, NH2, -NHC 1-4 Alkyl, -N(C 1-4 Alkyl)2, -NHC 3-10 Cycloalkyl, C 1-4 Alkoxy, C 1-6 Alkyl, C 3-10 Cycloalkyl, 6-10 membered aryl, 5-10 membered heteroaryl containing 1-3 heteroatoms selected from N, O, S; In certain embodiments, R a1 Selected from OH, NH2, -NHC 1-2Alkyl, -N(C 1-2 Alkyl)2, -NHC 3-7 Cycloalkyl, C 1-2 Alkoxy, C 1-2 Alkyl, C 3-7 Cycloalkyl, phenyl, 8-10 membered bicyclic aryl, 5-6 membered heteroaryl containing 1-3 heteroatoms selected from N, O, S, or 8-10 membered bicyclic heteroaryl containing 1-3 heteroatoms selected from N, O, S; in certain embodiments, R a1 Selected from OH, NH2, -NHC 1-2 Alkyl, -N(C 1-2 Alkyl)2, -NHC 3-4 Cycloalkyl, C 1-2 Alkoxy, C 1-2 Alkyl, C 3-4 Cycloalkyl, phenyl, piperazinyl, piperidinyl, tetrahydropyranyl, morpholinyl, pyranyl, phenyl, pyrrolyl, furanyl, thienyl, imidazolyl, oxazolyl, thiazolyl, pyrazolyl, isoxazolyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl; In certain embodiments, R a1 Selected from C 1-2 Alkyl, C 3-7 Cycloalkyl, phenyl, piperazinyl, piperidinyl, tetrahydropyranyl, morpholinyl, pyranyl, phenyl, pyrrolyl, furanyl, thienyl, imidazolyl, oxazolyl, thiazolyl, pyrazolyl, isoxazolyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl;

[0013] R 1 、R 2 、R 3 、R 4 、R 5 、R 6 、R 8 、R 9 Each independently selected from H, deuterium, halogen, OH, C 1-4 Alkyl, halogenated C 1-4 Alkyl, deuterated C 1-4 Alkyl, C 1-4 Alkoxy, halogenated C 1-4 Alkoxy, deuterated C 1-4 Alkoxy, C 3-6 Cycloalkyl, or R 1 and R 3 , or R 4 and R 5 , or R 3 and R 4 Together with the carbon atom to which it is attached, it forms C 3-6 Cycloalkyl, optionally further substituted by 1-3 groups selected from =O, Halogen, deuterium, CN, OH, C 1-3Alkyl, halogenated C 1-3 Alkyl, deuterated C 1-3 Alkyl, C 1-3 Alkoxy, halogenated C 1-3 Alkoxy, deuterated C 1-3 Alkoxy group substitution;

[0014] In some embodiments, R 1 、R 2 、R 3 、R 4 、R 6 、R 8 、R 9 Each independently selected from H, deuterium, halogen, OH, C 1-4 Alkyl, halogenated C 1-4 Alkyl, deuterated C 1-4 Alkyl, C 1-4 Alkoxy, halogenated C 1-4 Alkoxy, deuterated C 1-4 Alkoxy, C 3-6 Cycloalkyl, and R 1 and R 3 , or R 4 and R 5 , or R 3 and R 4 Together with the carbon atom to which it is connected, it forms a C3, C4, C5, or C6 cycloalkyl group, wherein the C3, C4, C5, or C6 cycloalkyl group is optionally further substituted by 1, 2, or 3 groups selected from ═O, Halogen, deuterium, CN, OH, C 1-3 Alkyl, halogenated C 1-3 Alkyl, deuterated C 1-3 Alkyl, C 1-3 Alkoxy, halogenated C 1-3 Alkoxy, deuterated C 1-3 Alkoxy group substitution;

[0015] In some embodiments, R 1 、R 2 、R 3 、R 4 、R 8 、R 9 Each independently selected from H, deuterium, C 1-2 Alkyl, halogenated C 1-2 Alkyl, C 3-6 Cycloalkyl;

[0016] In some embodiments, R 1 、R 2 、R 3 、R 4 、R 8 、R 9Each independently selected from H, deuterium, halogen, OH, C 1-4 Alkyl, halogenated C 1- 4-alkyl, deuterated C 1-4 Alkyl, C 1-4 Alkoxy, halogenated C 1-4 Alkoxy, deuterated C 1-4 Alkoxy, C 3-6 Cycloalkyl;

[0017] In some embodiments, R 1 、R 2 、R 3 、R 4 、R 5 、R 6 、R 8 、R 9 Each independently selected from H, deuterium, C 1-2 Alkyl, halogenated C 1-2 Alkyl, C 3-6 Cycloalkyl;

[0018] In some embodiments, R 1 、R 2 、R 5 、R 6 、R 8 、R 9 selected from H or deuterium;

[0019] In some embodiments, R 3 、R 4 Selected from H, deuterium, C 1-2 alkyl;

[0020] In some embodiments, R 1 and R 3 , or R 3 and R 4 Together with the carbon atom to which it is attached, it forms C 3-6 Cycloalkyl, optionally further substituted by 1-3 groups selected from =O, Halogen, deuterium, C 1-3 Alkyl, halogenated C 1-3 Alkyl, deuterated C 1-3 Alkyl, C 1-3 Alkoxy, halogenated C 1-3 Alkoxy, deuterated C 1-3 Alkoxy group substitution;

[0021] In some embodiments, R 1 and R 3 , or R 3 and R 4 Together with the carbon atom to which it is attached, it forms C 3-6Cycloalkyl, said cycloalkyl optionally further substituted by 1-3 groups selected from =O, Halogen, deuterium, C 1-3 Alkyl, halogenated C 1-3 Alkyl, deuterated C 1-3 Alkyl, C 1-3 Alkoxy, halogenated C 1-3 Alkoxy, deuterated C 1-3 Alkoxy group substitution;

[0022] In some embodiments, R 1 and R 3 , or R 3 and R 4 Together with the carbon atom to which it is attached, it forms C 3-6 Cycloalkyl, the cycloalkyl is optionally further substituted by 1-3 groups selected from Halogen, deuterium, C 1-3 Alkyl, halogenated C 1-3 Alkyl, deuterated C 1-3 Alkyl radical substitution;

[0023] In some embodiments, R 3 and R 4 Together with the carbon atom to which it is connected, it forms a C3, C4, C5, or C6 cycloalkyl group, wherein the C3, C4, C5, or C6 cycloalkyl group is optionally further substituted by 1, 2, or 3 groups selected from ═O, Halogen, deuterium, CN, OH, C 1-3 Alkyl, halogenated C 1-3 Alkyl, deuterated C 1-3 Alkyl, C 1-3 Alkoxy, halogenated C 1-3 Alkoxy, deuterated C 1-3 Alkoxy group substitution;

[0024] In some embodiments, R 5 and R 6 Together with the carbon atom to which it is attached, it forms =O, =CH2, =CF2 or a 5-10 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, S, which is optionally further substituted by 1-3 heteroatoms selected from =O, halogen, deuterium, CN, OH, C 1-3 Alkyl, halogenated C 1-3 Alkyl, deuterated C 1-3 Alkyl, C 1-3 Alkoxy, halogenated C 1-3 Alkoxy, deuterated C 1-3 Alkoxy group substitution;

[0025] In some embodiments, R 5 and R 6Together with the carbon atom to which it is attached, it forms =O, =CH2, =CF2 or a 5-6 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, S, which is optionally further substituted by 1-3 heteroatoms selected from halogen, deuterium, C 1-3 Alkyl, halogenated C 1-3 Alkyl, deuterated C 1-3 Alkyl radical substitution;

[0026] In some embodiments, R 1 and R 2 、R 3 and R 4 、R 5 and R 6 Together with the carbon atom to which it is attached, it forms =O, =CH2, =CF2 or a 5-10 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, S, which is optionally further substituted by 1-3 heteroatoms selected from =O, halogen, deuterium, CN, OH, C 1-3 Alkyl, halogenated C 1-3 Alkyl, deuterated C 1-3 Alkyl, C 1-3 Alkoxy, halogenated C 1-3 Alkoxy, deuterated C 1-3 Alkoxy group substitution;

[0027] In some embodiments, R 1 and R 2 、R 3 and R 4 Together with the carbon atom to which it is attached, it forms =O, =CH2, or =CF2;

[0028] In some embodiments, R 1 、R 2 、R 3 、R 4 、R 5 、R 6 Each independently selected from H, deuterium, halogen, OH, C 1-4 Alkyl, halogenated C 1- 4-alkyl, deuterated C 1-4 Alkyl, C 1-4 Alkoxy, halogenated C 1-4 Alkoxy, deuterated C 1-4 Alkoxy, C 3-6 Cycloalkyl, or R 1 and R 3 , or R 4 and R 5 Together with the carbon atom to which it is attached, it forms C 3-6 Cycloalkyl, optionally further substituted by 1-3 groups selected from =O, Halogen, deuterium, CN, OH, C1-3 Alkyl, halogenated C 1-3 Alkyl, deuterated C 1-3 Alkyl, C 1-3 Alkoxy, halogenated C 1-3 Alkoxy, deuterated C 1-3 In some embodiments, R 1 、R 2 、R 3 、R 4 、R 5 、R 6 Each independently selected from H, deuterium, F, Cl, OH, C 1-2 Alkyl, halogenated C 1-2 Alkyl, deuterated C 1-2 Alkyl, C 1-2 Alkoxy, halogenated C 1-2 Alkoxy, deuterated C 1-2 Alkoxy, C 3-4 Cycloalkyl, or R 1 and R 3 , or R 4 and R 5 Together with the carbon atom to which it is attached, it forms C 3-6 Cycloalkyl, optionally further substituted by 1, 2 or 3 groups selected from =O, F, Cl, deuterium, CN, OH, C 1-3 Alkyl, halogenated C 1-3 Alkyl, deuterated C 1-3 Alkyl, C 1-3 Alkoxy, halogenated C 1-3 Alkoxy, deuterated C 1-3 Alkoxy group substitution;

[0029] Alternatively, R 5 、R 6 Together with the carbon atom to which it is attached, it forms =O, =CH2, =CF2 or a 5-10 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, S, which is optionally further substituted by 1-3 heteroatoms selected from =O, halogen, deuterium, CN, OH, C 1-3 Alkyl, halogenated C 1-3 Alkyl, deuterated C 1-3 Alkyl, C 1-3 Alkoxy, halogenated C 1-3 Alkoxy, deuterated C 1-3 Alkoxy group substitution;

[0030] In some embodiments, alternatively, R 5 、R 6 Together with the carbon atom to which it is attached, it forms =O;

[0031] r is selected from 0, 1 or 2; in some embodiments, r is selected from 0, 1;

[0032] X is selected from N or CH; in some embodiments, X is selected from N; in some embodiments, X is selected from CH;

[0033] R 7 Selected from C 3-10 Cycloalkyl, 4-10 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, S, 6-10 membered aryl, 5-10 membered heteroaryl containing 1-3 heteroatoms selected from N, O, S, wherein the cycloalkyl, heterocycloalkyl, aryl, heteroaryl may be further substituted by 1-3 halogens, =O, deuterium, CN, OH, C 1-4 Alkyl, halogenated C 1-4 Alkyl, deuterated C 1-4 Alkyl, C 1-4 Alkoxy, halogenated C 1-4 Alkoxy, deuterated C 1-4 substituted with alkoxy, NH2, =CH2, and =CF2; in some embodiments, R 7 Selected from C 3-10 Cycloalkyl, wherein the cycloalkyl is optionally further substituted by 1-3 groups selected from halogen, =O, deuterium, CN, OH, C 1-4 Alkyl, halogenated C 1-4 Alkyl, deuterated C 1-4 Alkyl, C 1-4 Alkoxy, halogenated C 1-4 Alkoxy, deuterated C 1-4 Alkoxy, NH2, =CH2 and =CF2 group substitution;

[0034] And the compound of formula (I) is not the following structure:

[0035] In some embodiments, when R 5 and R 6 Together with the carbon atom to which it is attached, it forms =O, R 1 、R 2 、R 3 、R 4 、R 8 、R 9 When selected from H, R is not selected from the following structures:

[0036] Specifically, the first technical solution of the present invention provides a compound represented by formula (I) or formula (I-1), a stereoisomer or a pharmaceutically acceptable salt thereof,

[0037] in,

[0038] R is selected from -O-halogenated C 1-4 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, -O-(CH2) r -R a 、C 3-10 Cycloalkyl, 5-10 membered heteroaryl containing 1-3 heteroatoms selected from N, O, S, -C 1-4 Alkyl-R a 、-C 1-4 Alkyl-OR a 、-C 1-4 Alkyl-NR b R a 、-NR b -S(O)2-R a 、-NR b -S(O)2-NR b R a 、-O-NR b R a 、-NH-OR b 、-NH-halogenated C 1-4 Alkyl, -OC 1-4 Alkyl, C 1-6 Alkoxy, halogen, cyano, nitro, C 1-4 Alkyl, -NH-C 1-4 Alkyl, -N(C 1-4 Alkyl)2, -NH-C 3-10 Cycloalkyl, -NHC(O)C 1-4 Alkyl, 4-10 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, S, -CO-C 1-4 Alkyl, -CO-R a 、-S(O)-R a 、-S(O)2-R a The alkyl, alkoxy, alkenyl, alkynyl, heteroaryl, cycloalkyl, heterocycloalkyl may be further substituted by 1-3 halogen, D, CN, OH, C 1-4 Alkyl, halogenated C 1-4 Alkyl, deuterated C 1-4 Alkyl, C 1-4 Alkoxy, halogenated C 1-4 Alkoxy, deuterated C 1-4 Alkoxy, C 3-10 substituted by cycloalkyl, 4-6 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, S, phenyl and NH2;

[0039] R a Selected from CN, C 3-10Cycloalkyl, 4-10 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, S, 6-10 membered aryl, 5-10 membered heteroaryl containing 1-3 heteroatoms selected from N, O, S, -C(O)-R a1 The cycloalkyl, heterocycloalkyl, aryl, heteroaryl groups are optionally further substituted by 1-3 groups selected from halogen, =O, deuterium, CN, OH, C 1-4 Alkyl, halogenated C 1-4 Alkyl, deuterated C 1-4 Alkyl, C 1-4 Alkoxy, halogenated C 1-4 Alkoxy, deuterated C 1-4 Alkoxy and NH2 group substitution;

[0040] Each R b Selected from H, deuterium, C 1-4 Alkyl, C 3-6 Cycloalkyl, halogenated C 1-4 Alkyl, deuterated C 1-4 alkyl;

[0041] R a1 Selected from OH, NH2, -NHC 1-4 Alkyl, -N(C 1-4 Alkyl)2, -NHC 3-10 Cycloalkyl, C 1-4 Alkoxy, C 1-6 Alkyl, C 3-10 Cycloalkyl, 6-10 membered aryl, 5-10 membered heteroaryl containing 1-3 heteroatoms selected from N, O, S;

[0042] R 1 、R 2 、R 3 、R 4 、R 5 、R 6 、R 8 、R 9 Each independently selected from H, deuterium, halogen, OH, C 1-4 Alkyl, halogenated C 1-4 Alkyl, deuterated C 1-4 Alkyl, C 1-4 Alkoxy, halogenated C 1-4 Alkoxy, deuterated C 1-4 Alkoxy, C 3-6 Cycloalkyl, or R 1 and R 3 , or R 4 and R 5 , or R 3 and R 4 Together with the carbon atom to which it is attached, it forms C 3-6Cycloalkyl, optionally further substituted by 1-3 groups selected from =O, Halogen, deuterium, CN, OH, C 1-3 Alkyl, halogenated C 1-3 Alkyl, deuterated C 1-3 Alkyl, C 1-3 Alkoxy, halogenated C 1-3 Alkoxy, deuterated C 1-3 Alkoxy group substitution;

[0043] Alternatively, R 1 and R 2 、R 3 and R 4 、R 5 and R 6 Together with the carbon atom to which it is attached, it forms =O, =CH2, =CF2 or a 5-10 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, S, which is optionally further substituted by 1-3 heteroatoms selected from =O, halogen, deuterium, CN, OH, C 1-3 Alkyl, halogenated C 1-3 Alkyl, deuterated C 1-3 Alkyl, C 1-3 Alkoxy, halogenated C 1-3 Alkoxy, deuterated C 1-3 Alkoxy group substitution;

[0044] In some embodiments, R 1 、R 2 、R 3 、R 4 、R 5 、R 6 Each independently selected from H, deuterium, halogen, OH, C 1-4 Alkyl, halogenated C 1- 4-alkyl, deuterated C 1-4 Alkyl, C 1-4 Alkoxy, halogenated C 1-4 Alkoxy, deuterated C 1-4 Alkoxy, C 3-6 Cycloalkyl, or R 1 and R 3 , or R 4 and R 5 Together with the carbon atom to which it is attached, it forms C 3-6 Cycloalkyl, optionally further substituted by 1-3 groups selected from =O, Halogen, deuterium, CN, OH, C 1-3 Alkyl, halogenated C 1-3 Alkyl, deuterated C 1-3 Alkyl, C 1-3 Alkoxy, halogenated C 1-3 Alkoxy, deuterated C 1-3Alkoxy group substitution;

[0045] Alternatively, R 5 、R 6 Together with the carbon atom to which it is attached, it forms =O, =CH2, =CF2 or a 5-10 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, S, which is optionally further substituted by 1-3 heteroatoms selected from =O, halogen, deuterium, CN, OH, C 1-3 Alkyl, halogenated C 1-3 Alkyl, deuterated C 1-3 Alkyl, C 1-3 Alkoxy, halogenated C 1-3 Alkoxy, deuterated C 1-3 Alkoxy group substitution;

[0046] X is selected from N or CH;

[0047] R 7 Selected from C 3-10 Cycloalkyl, 4-10 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, S, 6-10 membered aryl, 5-10 membered heteroaryl containing 1-3 heteroatoms selected from N, O, S, wherein the cycloalkyl, heterocycloalkyl, aryl, heteroaryl may be further substituted by 1-3 halogens, =O, deuterium, CN, OH, C 1-4 Alkyl, halogenated C 1-4 Alkyl, deuterated C 1-4 Alkyl, C 1-4 Alkoxy, halogenated C 1-4 Alkoxy, deuterated C 1-4 Alkoxy, NH2, =CH2 and =CF2 group substitution;

[0048] r is selected from 0, 1 or 2;

[0049] And the compound of formula (I) is not the following structure:

[0050] The second technical solution of the present invention provides a compound represented by formula (I), a stereoisomer or a pharmaceutically acceptable salt thereof,

[0051] in,

[0052] R is selected from -O-halogenated C 1-4 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, -O-(CH2) r -R a 、C 3-10 Cycloalkyl, 5-10 membered heteroaryl containing 1-3 heteroatoms selected from N, O, S, -C 1-4 Alkyl-Ra 、-C 1-4 Alkyl-OR a 、-C 1-4 Alkyl-NR b R a 、-NR b -S(O)2-R a 、-NR b -S(O)2-NR b R a 、-O-NR b R a 、-NH-OR b 、-NH-halogenated C 1-4 Alkyl, -OC 1-4 Alkyl, C 1-6 Alkoxy, halogen, cyano, nitro, C 1-4 Alkyl, -NH-C 1-4 Alkyl, -N(C 1-4 Alkyl)2, -NH-C 3-10 Cycloalkyl, -NHC(O)C 1-4 Alkyl or 4-10 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, S, wherein the alkyl, alkoxy, alkenyl, alkynyl, heteroaryl, cycloalkyl, heterocycloalkyl is optionally further substituted by 1-3 heteroatoms selected from halogen, D, CN, OH, C 1-4 Alkyl, halogenated C 1-4 Alkyl, deuterated C 1-4 Alkyl, C 1-4 Alkoxy, halogenated C 1-4 Alkoxy, deuterated C 1-4 Alkoxy, C 3-10 Cycloalkyl, 4-6 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, S, phenyl, and NH2;

[0053] R a Selected from CN, C 3-10 Cycloalkyl, 4-10 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, S, 6-10 membered aryl, 5-10 membered heteroaryl containing 1-3 heteroatoms selected from N, O, S, -C(O)-R a1 The cycloalkyl, heterocycloalkyl, aryl, heteroaryl groups are optionally further substituted by 1-3 groups selected from halogen, =O, deuterium, CN, OH, C 1-4 Alkyl, halogenated C 1-4 Alkyl, deuterated C 1-4 Alkyl, C 1-4 Alkoxy, halogenated C 1-4 Alkoxy, deuterated C 1-4 Alkoxy and NH2 group substitution;

[0054] Each Rb Selected from H, deuterium, C 1-4 Alkyl, C 3-6 Cycloalkyl, halogenated C 1-4 Alkyl, deuterated C 1-4 alkyl;

[0055] R a1 Selected from OH, NH2, -NHC 1-4 Alkyl, -N(C 1-4 Alkyl)2, -NHC 3-10 Cycloalkyl, C 1-4 Alkoxy, C 1-6 Alkyl, C 3-10 Cycloalkyl, 6-10 membered aryl, 5-10 membered heteroaryl containing 1-3 heteroatoms selected from N, O, S;

[0056] R 1 、R 2 、R 3 、R 4 、R 5 、R 6 Each independently selected from H, deuterium, halogen, OH, C 1-4 Alkyl, halogenated C 1-4 Alkyl, deuterated C 1-4 Alkyl, C 1-4 Alkoxy, halogenated C 1-4 Alkoxy, deuterated C 1-4 Alkoxy, C 3-6 Cycloalkyl, or R 1 and R 3 , or R 4 and R 5 Together with the carbon atom to which it is attached, it forms C 3-6 Cycloalkyl, optionally further substituted by 1-3 groups selected from =O, Halogen, deuterium, CN, OH, C 1-3 Alkyl, halogenated C 1-3 Alkyl, deuterated C 1-3 Alkyl, C 1-3 Alkoxy, halogenated C 1-3 Alkoxy, deuterated C 1-3 Alkoxy group substitution;

[0057] Alternatively, R 5 、R 6 Together with the carbon atom to which it is attached, it forms =O;

[0058] r is selected from 0, 1 or 2;

[0059] The condition is that the compound of formula (I) satisfies the following conditions:

[0060] (1)R 1 and R3 , or R 4 and R 5 Together with the carbon atom to which it is attached, it forms C 3-6 Cycloalkyl, optionally further substituted by 1-3 groups selected from =O, Halogen, deuterium, CN, OH, C 1-3 Alkyl, halogenated C 1-3 Alkyl, deuterated C 1-3 Alkyl, C 1-3 Alkoxy, halogenated C 1-3 Alkoxy, deuterated C 1-3 Alkoxy group substitution; or

[0061] (2) When R 5 、R 6 When R forms =O together with the carbon atom to which it is attached, 1 、R 2 、R 3 、R 4 、R 5 、R 6 wherein at least one group is selected from deuterium;

[0062] And the compound of formula (I) is not the following structure:

[0063] The third technical solution of the present invention is the compound of the present invention, its stereoisomer or pharmaceutically acceptable salt, wherein:

[0064] R is selected from -CF3, -CH2CH3, -CH2CH2CH3, -COCH3, -COCF3, In some embodiments, R is selected from -CF3, -CH2CH3, -CH2CH2CH3,

[0065] The fourth technical solution of the present invention is the compound of the present invention, its stereoisomers or pharmaceutically acceptable salts,

[0066] R 1 、R 2 、R 3 、R 4 、R 6 、R 8 、R 9 Each independently selected from H, deuterium, halogen, OH, C 1-4 Alkyl, halogenated C 1-4 Alkyl, deuterated C 1-4 alkyl, C 1-4 Alkoxy, halogenated C 1-4 Alkoxy, deuterated C 1-4 Alkoxy, C 3-6 Cycloalkyl, and R 1 and R 3 , or R 4 and R 5 , or R 3 and R 4 Together with the carbon atom to which it is connected, it forms a C3, C4, C5, or C6 cycloalkyl group, wherein the C3, C4, C5, or C6 cycloalkyl group is optionally further substituted by 1, 2, or 3 groups selected from ═O, Halogen, deuterium, CN, OH, C 1-3 Alkyl, halogenated C 1-3 Alkyl, deuterated C 1-3 Alkyl, C 1-3 Alkoxy, halogenated C 1-3 Alkoxy, deuterated C 1-3 Alkoxy group substitution;

[0067] In some embodiments, R 2 、R 4 、R 6 Each independently selected from H, deuterium, halogen, OH, C 1-4 Alkyl, halogenated C 1-4 Alkyl, deuterated C 1-4 Alkyl, C 1-4 Alkoxy, halogenated C 1-4 Alkoxy, deuterated C 1-4 Alkoxy, C 3-6 Cycloalkyl, and R 1 and R 3 , or R 4 and R 5 Together with the carbon atom to which it is connected, it forms a C3, C4, or C5 cycloalkyl group, wherein the C3, C4, or C5 cycloalkyl group is optionally further substituted by 1, 2, or 3 groups selected from ═O, Halogen, deuterium, CN, OH, C 1-3 Alkyl, halogenated C 1-3 Alkyl, deuterated C 1-3 Alkyl, C 1-3 Alkoxy, halogenated C 1-3 Alkoxy, deuterated C 1-3 Alkoxy groups are substituted.

[0068] The fifth technical solution of the present invention is the compound of the present invention, its stereoisomer or pharmaceutically acceptable salt,

[0069] R 1 、R 2 、R 3 、R4 、R 8 、R 9 Each independently selected from H, deuterium, halogen, OH, C 1-4 Alkyl, halogenated C 1-4 Alkyl, deuterated C 1-4 Alkyl, C 1-4 Alkoxy, halogenated C 1-4 Alkoxy, deuterated C 1-4 Alkoxy, C 3-6 Cycloalkyl, or R 3 and R 4 Together with the carbon atom to which it is connected, it forms a C3, C4, C5, or C6 cycloalkyl group, wherein the C3, C4, C5, or C6 cycloalkyl group is optionally further substituted by 1, 2, or 3 groups selected from ═O, Halogen, deuterium, CN, OH, C 1-3 Alkyl, halogenated C 1-3 Alkyl, deuterated C 1-3 Alkyl, C 1-3 Alkoxy, halogenated C 1-3 Alkoxy, deuterated C 1-3 substituted with an alkoxy group; and R 5 、R 6 Together with the carbon atom to which it is attached, it forms =O, =CH2, =CF2 or a 5-10 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, S, which is optionally further substituted by 1-3 heteroatoms selected from =O, halogen, deuterium, CN, OH, C 1-3 Alkyl, halogenated C 1-3 Alkyl, deuterated C 1-3 Alkyl, C 1-3 Alkoxy, halogenated C 1-3 Alkoxy, deuterated C 1-3 Alkoxy group substitution;

[0070] In some embodiments, R 1 、R 2 、R 3 、R 4 Each independently selected from H, deuterium, halogen, OH, C 1-4 Alkyl, halogenated C 1-4 Alkyl, deuterated C 1-4 Alkyl, C 1-4 Alkoxy, halogenated C 1-4 Alkoxy, deuterated C 1-4 Alkoxy, C 3-6 Cycloalkyl, and R 5 、R 6Together with the carbon atom to which it is attached, it forms =O, =CH2, =CF2 or a 5-10 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, S, which is optionally further substituted by 1-3 heteroatoms selected from =O, halogen, deuterium, CN, OH, C 1-3 Alkyl, halogenated C 1-3 Alkyl, deuterated C 1-3 Alkyl, C 1-3 Alkoxy, halogenated C 1-3 Alkoxy, deuterated C 1-3 Alkoxy groups are substituted.

[0071] The sixth technical solution of the present invention is that the compound of the present invention, its stereoisomers or pharmaceutically acceptable salts have the structure of formula (II) or (II-1):

[0072] Wherein, R is selected from -O-halogenated C 1-4 Alkyl, -O- containing 1-3 heteroatoms selected from N, O, S 4-10 membered heterocycloalkyl, the alkyl, heterocycloalkyl optionally further 1-3 selected from halogen, D, CN, OH, C 1-4 Alkyl, halogenated C 1-4 Alkyl, deuterated C 1-4 Alkyl, C 1-4 Alkoxy, halogenated C 1-4 Alkoxy, deuterated C 1-4 Alkoxy and NH2 groups are substituted.

[0073] The seventh technical solution of the present invention is that the compound of the present invention, its stereoisomers or pharmaceutically acceptable salts have the structure of formula (I-2):

[0074] R is selected from -O-halogenated C 1-4 Alkyl, -O- containing 1-3 heteroatoms selected from N, O, S 4-10 membered heterocycloalkyl, the alkyl, heterocycloalkyl optionally further 1-3 selected from halogen, D, CN, OH, C 1-4 Alkyl, halogenated C 1-4 Alkyl, deuterated C 1-4 Alkyl, C 1-4 Alkoxy, halogenated C 1-4 Alkoxy, deuterated C 1-4 Alkoxy and NH2 group substitution;

[0075] R 7 Selected from C 3-10 Cycloalkyl, wherein the cycloalkyl is optionally further substituted by 1-3 groups selected from halogen, =O, deuterium, CN, OH, C 1-4 Alkyl, halogenated C 1-4 Alkyl, deuterated C1-4 Alkyl, C 1-4 Alkoxy, halogenated C 1-4 Alkoxy, deuterated C 1-4 The alkyl group is substituted with alkoxy, NH2, =CH2 and =CF2.

[0076] The eighth technical solution of the present invention, the compound of formula (II), its stereoisomers or pharmaceutically acceptable salts, wherein

[0077] R is selected from

[0078] The ninth technical solution of the present invention is the compound of formula (I), its stereoisomers or pharmaceutically acceptable salts, wherein:

[0079] R is selected from -COCH3, -COCF3, -COCH3CF3,

[0080] R 1 、R 2 、R 3 、R 4 、R 5 、R 6 、R 8 、R 9 Each independently selected from H, deuterium, C 1-2 Alkyl, halogenated C 1-2 Alkyl, C 3-6 Cycloalkyl;

[0081] or R 1 and R 3 , or R 3 and R 4 Together with the carbon atom to which it is attached, it forms C 3-6 Cycloalkyl, said cycloalkyl optionally further substituted by 1-3 groups selected from =O, Halogen, deuterium, C 1-3 Alkyl, halogenated C 1-3 Alkyl, deuterated C 1-3 Alkyl, C 1-3 Alkoxy, halogenated C 1-3 Alkoxy, deuterated C 1-3 Alkoxy group substitution;

[0082] Alternatively, R 5 and R 6Together with the carbon atom to which it is attached, it forms =O, =CH2, =CF2 or a 5-10 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, S, which is optionally further substituted by 1-3 heteroatoms selected from =O, halogen, deuterium, CN, OH, C 1-3 Alkyl, halogenated C 1-3 Alkyl, deuterated C 1-3 Alkyl, C 1-3 Alkoxy, halogenated C 1-3 Alkoxy, deuterated C 1-3 Alkoxy group substitution;

[0083] Alternatively, R 1 and R 2 、R 3 and R 4 Together with the carbon atom to which it is attached, it forms =O, =CH2, or =CF2;

[0084] The condition is that when R 5 and R 6 Together with the carbon atom to which it is attached, it forms =O, R 1 、R 2 、R 3 、R 4 、R 8 、R 9 When selected from H, R is not selected from the following structures:

[0085] The tenth technical solution of the present invention is the compound of formula (I), its stereoisomers or pharmaceutically acceptable salts, wherein,

[0086] R 1 、R 2 、R 5 、R 6 、R 8 、R 9 selected from H or deuterium;

[0087] R 3 、R 4 Selected from H, deuterium, C 1-2 alkyl;

[0088] or R 1 and R 3 , or R 3 and R 4 Together with the carbon atom to which it is attached, it forms C 3-6 Cycloalkyl, the cycloalkyl is optionally further substituted by 1-3 groups selected from Halogen, deuterium, C 1-3 Alkyl, halogenated C 1-3 Alkyl, deuterated C 1-3Alkyl radical substitution;

[0089] Alternatively, R 5 and R 6 Together with the carbon atom to which it is attached, it forms =O, =CH2, =CF2 or a 5-6 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, S, which is optionally further substituted by 1-3 heteroatoms selected from halogen, deuterium, C 1-3 Alkyl, halogenated C 1-3 Alkyl, deuterated C 1-3 Alkyl groups are substituted.

[0090] The compounds of formula (I), formula (I-1), formula (II), formula (II-1), formula (I-1), formula (I-2), and formula (I-3) of the present invention, their stereoisomers or pharmaceutically acceptable salts, are selected from one of the structures in Table 1 or Table 2:

[0091] Table 1:

[0092] Table 2:

[0093] Secondly, the present invention also provides a pharmaceutical composition or pharmaceutical preparation, which contains the compound described in any one of the aforementioned technical solutions, its stereoisomers, solvates, deuterated substances, or pharmaceutically acceptable salts, and pharmaceutically acceptable carriers and / or excipients.

[0094] Furthermore, the pharmaceutical composition or pharmaceutical preparation comprises 1-1500 mg of the compound described in any of the aforementioned technical solutions, its stereoisomers or pharmaceutically acceptable salts, and a pharmaceutically acceptable carrier and / or excipient.

[0095] Furthermore, the present invention also provides the use of a compound according to any of the preceding embodiments, or a stereoisomer, solvate, deuterated form, or pharmaceutically acceptable salt, or a pharmaceutical composition or formulation thereof, in the preparation of a medicament for treating or preventing Myosin II-mediated diseases. Furthermore, Myosin II-mediated diseases include, but are not limited to, muscular dystrophy.

[0096] The present invention also provides a method for treating a disease in a mammal, comprising administering to a subject a therapeutically effective amount of a compound according to any one of the aforementioned technical solutions, or a stereoisomer or a pharmaceutically acceptable salt thereof, wherein the therapeutically effective amount is preferably 1-1500 mg, and the disease is preferably muscular dystrophy.

[0097] The present invention also provides a method for treating a disease in a mammal, comprising administering to the mammal a therapeutically effective amount of the compound, stereoisomer, or pharmaceutically acceptable salt, or pharmaceutical composition, or pharmaceutical preparation of the present invention. In some embodiments, the mammal of the present invention includes a human.

[0098] As used herein, an "effective amount" or "therapeutically effective amount" refers to the administration of a sufficient amount of a compound disclosed herein to alleviate, to some extent, one or more symptoms of the disease or condition being treated. In some embodiments, the result is a reduction and / or alleviation of the signs, symptoms, or causes of a disease, or any other desired alteration in a biological system. For example, an "effective amount" for therapeutic uses is the amount of a compound disclosed herein required to provide a clinically significant reduction in disease symptoms. Examples of therapeutically effective amounts include, but are not limited to, 1-1500 mg, 1-1400 mg, 1-1300 mg, 1-1200 mg, 1-1000 mg, 1-900 mg, 1-800 mg, 1-700 mg, 1-600 mg, 1-500 mg, 1-400 mg, 1-300 mg, 1-250 mg, 1-200 mg, 1-150 mg, 1-125 mg, 1-100 mg, 1-80 mg, 1-60 mg, 1-50 mg, 1-40 mg, 1-25 mg, 1- 20mg, 5-1500mg, 5-1000mg, 5-900mg, 5-800mg, 5-700mg, 5-600mg, 5-500mg, 5-400mg, 5-300mg, 5-250mg, 5-200mg, 5 -150mg, 5-125mg, 5-100mg, 5-90mg, 5-70mg, 5-80mg, 5-60mg, 5-50mg, 5-40mg, 5-30mg, 5-25mg, 5-20mg, 10-1500mg, 10-1000mg, 10-900mg, 10-800mg, 10-700mg, 10-600mg, 10-500mg, 10-450mg, 10-400mg, 10-300mg, 10-250mg, 10-20 0mg, 10-150mg, 10-125mg, 10-100mg, 10-90mg, 10-80mg, 10-70mg, 10-60mg, 10-50mg, 10-40mg, 10-30mg, 10-20mg; 2 0-1500mg, 20-1000mg, 20-900mg, 20-800mg, 20-700mg, 20-600mg, 20-500mg, 20-400mg, 20-350mg, 20-300mg, 20-25 0mg, 20-200mg, 20-150mg, 20-125mg, 20-100mg, 20-90mg, 20-80mg, 20-70mg, 20-60mg, 20-50mg, 20-40mg, 20-30mg;50-1500mg, 50-1000mg, 50-900mg, 50-800mg, 50-700mg, 50-600mg, 50-500mg, 50-400mg, 50-300mg, 50-250mg, 50-200mg, 50-150mg, 50-125mg, 5 0-100mg; 100-1500mg, 100-1000mg, 100-900mg, 100-800mg, 100-700mg, 100-600mg, 100-500mg, 100-400mg, 100-300mg, 100-250mg, 100-200mg. ;

[0099] The present invention relates to a pharmaceutical composition or pharmaceutical preparation comprising a therapeutically effective amount of a compound, stereoisomer, or pharmaceutically acceptable salt of the present invention and a carrier and / or excipient. The pharmaceutical composition may be in the form of a unit dosage form (the amount of the active ingredient in a unit dosage form is also referred to as the "drug strength"). In some embodiments, the pharmaceutical composition includes but is not limited to 1-1500 mg, 5-1000 mg, 10-800 mg, 20-600 mg, 25-500 mg, 40-200 mg, 50-100 mg, 1 mg, 1.25 mg, 2.5 mg, 5 mg, 10 mg, 12.5 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 55 mg, 60 mg, 65 mg, 70 mg, 75 mg, 80 mg, 85 mg, 90 mg, 95 mg, 100 mg, 110 mg, 120 mg, 125 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 , 750 mg, 800 mg, 850 mg, 900 mg, 950 mg, 1000 mg, 1100 mg, 1200 mg, 1300 mg, 1400 mg, 1500 mg of a compound of the present invention, or a stereoisomer or a pharmaceutically acceptable salt thereof.

[0100] A method for treating a disease in a mammal, comprising administering to a subject a therapeutically effective amount of a compound of the present invention, a stereoisomer or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier and / or excipient, wherein the therapeutically effective amount is preferably 1-1500 mg, and the disease is preferably muscular dystrophy.

[0101] A method for treating a disease in a mammal, comprising administering a compound of the present invention, a stereoisomer or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier and / or excipient to a subject at a daily dose of 1-1500 mg / day, wherein the daily dose can be a single dose or divided doses. In some embodiments, the daily dose includes but is not limited to 10-1500 mg / day, 20-1500 mg / day, 25-1500 mg / day, 50-1500 mg / day, 75-1500 mg / day, 100-1500 mg / day, 200-1500 mg / day, 10-1000 mg / day, 20-1000 mg / day, 25-1000 mg / day, 50-1000 mg / day, 75-1000 mg / day, 100 -1000 mg / day, 200-1000 mg / day, 25-800 mg / day, 50-800 mg / day, 100-800 mg / day, 200-800 mg / day, 25-400 mg / day, 50-400 mg / day, 100-400 mg / day, 200-400 mg / day, in some embodiments, daily doses include but are not limited to 1 mg / day, 5 mg / day, 10 mg / day, 20 mg / day, 25 mg / day, 50 mg / day, 75 mg / day, 100 mg / day, 125 mg / day, 150 mg / day, 200 mg / day, 400 mg / day, 600 mg / day, 800 mg / day, 1000 mg / day, 1200 mg / day, 1400 mg / day, 1500 mg / day.

[0102] The present invention relates to a kit which may include a composition in single-dose or multi-dose form, wherein the kit contains a compound of the present invention or a stereoisomer or a pharmaceutically acceptable salt, and the amount of the compound of the present invention or the stereoisomer or the pharmaceutically acceptable salt is the same as that in the above-mentioned pharmaceutical composition.

[0103] The amount of the compound according to the invention or the stereoisomer or the pharmaceutically acceptable salt is in each case calculated as the free base.

[0104] "Preparation specifications" refers to the weight of the main drug contained in each vial, tablet or other unit preparation.

[0105] Synthesis route

[0106] Those skilled in the art can prepare the compounds of the present invention by combining known organic synthesis techniques, using commercially available chemicals and / or compounds described in the chemical literature as starting materials. "Commercially available chemicals" are obtained from reputable commercial sources, including suppliers such as Titan Technology, Anage Chemical, Shanghai Demo, Chengdu Kelon Chemical, Shaoyuan Chemical Technology, Nanjing Yaoshi, WuXi AppTec, and J&K Technology.

[0107] Specific and similar reactants can be selectively identified by indexes of known chemical substances prepared by the American Chemical Society's Chemical Abstracts Service, which are available in most public and university libraries and online. Chemicals that are known but not commercially available in the catalog are optionally prepared by custom chemical synthesis facilities, many of which standard chemical supply facilities (e.g., those listed above) offer custom synthesis services.

[0108] the term

[0109] Unless otherwise specified in the present invention, the terms of the present invention have the following meanings:

[0110] The carbon, hydrogen, oxygen, sulfur, nitrogen or halogen involved in the groups and compounds of the present invention include their isotopes, and the carbon, hydrogen, oxygen, sulfur, nitrogen or halogen involved in the groups and compounds of the present invention are optionally further replaced by one or more of their corresponding isotopes, wherein the isotopes of carbon include 12 C. 13 C and 14 C, hydrogen isotopes include protium (H), deuterium (deuterium, also known as heavy hydrogen), tritium (T, also known as super tritium), oxygen isotopes include 16 O. 17 O and 18 O, sulfur isotopes include 32 S. 33 S. 34 S and 36 S, nitrogen isotopes include 14 N and 15 N, an isotope of fluorine 19 F, chlorine isotopes include 35 Cl and 37 Isotopes of Cl, bromine include 79 Br and 81 Br.

[0111] "Halogen" herein refers to F, Cl, Br, I, or isotopes thereof.

[0112] "Halo" or "halogen-substituted" refers to substitution with one or more halogens selected from F, Cl, Br, I, or isotopes thereof. The upper limit of the number of halogen substituents is equal to the sum of the number of hydrogen atoms that can be replaced by the substituted group. Unless otherwise specified, the number of halogen substituents is any integer between 1 and the upper limit. When the number of halogen substituents is greater than 1, they may be the same or different halogens. Typical examples include 1-5 halogen substitutions, 1-3 halogen substitutions, 1-2 halogen substitutions, and 1 halogen substitution.

[0113] "Deuterium" refers to the isotope of hydrogen (H) and is synonymous with "D".

[0114] "Deuterated" or "deuterated compound" refers to a situation where a hydrogen atom on an alkyl, cycloalkyl, alkylene, aryl, heteroaryl, thiol, heterocycloalkyl, alkenyl, alkynyl or other group is replaced by at least one deuterium atom. The upper limit of the number of deuterated groups is equal to the sum of the number of replaceable hydrogen atoms in the substituted group. Unless otherwise specified, the number of deuterated groups is any integer between 1 and the upper limit, for example, 1-20 deuterium atoms, 1-10 deuterium atoms, 1-6 deuterium atoms, 1-3 deuterium atoms, 1-2 deuterium atoms or 1 deuterium atom.

[0115] “C x-y " group refers to a group containing x to y carbon atoms, such as "C 1-6 "Alkyl" refers to an alkyl group containing 1 to 6 carbon atoms.

[0116] "Alkyl" refers to a monovalent straight-chain or branched saturated aliphatic hydrocarbon group. Typically, it is an alkyl group having 1 to 20 carbon atoms, or an alkyl group having 1 to 8 carbon atoms, or an alkyl group having 1 to 6 carbon atoms, or an alkyl group having 1 to 4 carbon atoms. Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, neobutyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, and the like. The alkyl group may be further substituted with a substituent.

[0117] "Alkylene" refers to a divalent straight-chain or branched saturated alkyl group. Examples of alkylene groups include, but are not limited to, methylene, ethylene, and the like.

[0118] "Haloalkyl" or "haloC x-y"Alkyl" refers to a situation where one or more hydrogen atoms in the alkyl group are replaced by one or more halogen atoms (such as fluorine, chlorine, bromine, iodine or their isotopes). The upper limit of the number of halogen substituents is equal to the sum of the number of hydrogen atoms that can be replaced in the alkyl group. Unless otherwise specified, the number of halogen substituents is any integer between 1 and the upper limit. Usually, the alkyl group is substituted by 1-5 halogens, or 1-3 halogens, or 1-2 halogens or 1 halogen. When the number of halogen substituents is greater than 1, they can be the same or different halogens. Specific examples include but are not limited to -CF3, -CH2Cl, -CH2CF3, -CCl2, CF3, etc.

[0119] "Alkoxy" or "alkyloxy" refers to an -O-alkyl group. For example, -OC 1-8 Alkyl, -OC 1-6 Alkyl, -OC 1-4 Alkyl or -OC 1-2 Specific non-limiting examples include methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, sec-butoxy, tert-butoxy, n-pentoxy, n-hexyloxy, cyclopropyloxy, and cyclobutyloxy, and the like; the alkoxy groups may be optionally substituted with substituents.

[0120] "Haloalkoxy" refers to an -O-haloalkyl group. For example, -O-haloC 1-8 Alkyl, -O-halogenated C 1-6 Alkyl, -O-halogenated C 1-4 Alkyl or -O-halogenated C 1-2 Alkyl; the upper limit of the number of halogen substituents is equal to the sum of the number of hydrogen atoms that can be substituted by the substituted group. Unless otherwise specified, the number of halogen substituents is any integer between 1 and the upper limit, preferably 1-5 halogen substitutions, 1-3 halogen substitutions, 1-2 halogen substitutions, or 1 halogen substitution; when the number of halogen substituents is greater than 1, they may be the same or different halogen substitutions; non-limiting examples include monofluoromethoxy, difluoromethoxy, trifluoromethoxy, difluoroethyloxy, and the like.

[0121] "Alkenyl" refers to a straight or branched chain hydrocarbon group containing at least one carbon-carbon double bond (C=C), typically containing 2 to 18 carbon atoms, such as 2 to 8 carbon atoms, further such as 2 to 6 carbon atoms, and further such as 2 to 4 carbon atoms, examples of which include but are not limited to vinyl, allyl, 1-propenyl, 2-propenyl, 1-butenyl, 2-butenyl, 3-butenyl, 1-pentenyl, 2-pentenyl, 3-pentenyl, 4-pentenyl, 1-methyl-1-butenyl, 2-methyl-1-butenyl, 2-methyl-2-butenyl, 2-methyl-3-butenyl, 2-methyl-4-butenyl, 2-methyl-5-butenyl, 2-methyl-6-butenyl, 2-methyl-7-butenyl, 2-methyl-8-butenyl, 2-methyl-9-butenyl, 2-methyl-1-butenyl, 2-methyl-2-butenyl, 2-methyl-3 ... -methyl-3-butenyl, 1-hexenyl, 2-hexenyl, 3-hexenyl, 4-hexenyl, 5-hexenyl, 1-methyl-1-pentenyl, 2-methyl-1-pentenyl, 1-heptenyl, 2-heptenyl, 3-heptenyl, 4-heptenyl, 1-octenyl, 3-octenyl, 1-nonenyl, 3-nonenyl, 1-decenyl, 4-decenyl, 1,3-butadiene, 1,3-pentadiene, 1,4-pentadiene and 1,4-hexadiene, etc.; the alkenyl group may be optionally further substituted with a substituent.

[0122] "Alkynyl" refers to a straight-chain or branched hydrocarbon group containing at least one carbon-carbon triple bond (C≡C), typically containing 2 to 18 carbon atoms, further containing 2 to 8 carbon atoms, further containing 2 to 6 carbon atoms, and further containing 2 to 4 carbon atoms. Examples include, but are not limited to, ethynyl, 1-propynyl, 2-propynyl, butynyl, 2-butynyl, 3-butynyl, 1-methyl-2-propynyl, 4-pentynyl, 3-pentynyl, 1-methyl-2-butynyl, 2-hexynyl, 3-hexynyl, 2-heptynyl, 3-heptynyl, 4-heptynyl, 3-octynyl, 3-nonynyl, and 4-decynyl. The alkynyl group may be optionally substituted with a substituent.

[0123] "Cycloalkyl" refers to a saturated or partially unsaturated, non-aromatic carbocyclic hydrocarbon group containing no ring heteroatoms. Cycloalkyl can be monocyclic, bicyclic or polycyclic. Bicyclic or polycyclic rings can be cyclic, spirocyclic, bridged or a combination thereof. Bicyclic or polycyclic rings can include one or more aromatic rings, but the ring system as a whole does not have aromaticity. The attachment site can be on the aromatic ring or on the non-aromatic ring. Usually, the cycloalkyl group contains 3 to 20 carbon atoms, further contains 3-8 carbon atoms, and further contains 3-6 carbon atoms; when it is a monocyclic cycloalkyl group, it contains 3-15 carbon atoms, or 3-10 carbon atoms, or 3-8 carbon atoms, or 3-6 carbon atoms; when it is a bicyclic or polycyclic cycloalkyl group, it contains 5-12 carbon atoms, or 5-11 carbon atoms, or 6-10 carbon atoms; non-limiting examples include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, butenyl, cyclopentenyl, cyclohexenyl, The cycloalkyl group may be optionally substituted with a substituent.

[0124] "Aryl" refers to a carbon ring with aromaticity and no heteroatoms, including monocyclic aromatic groups and fused aromatic groups, and aromatic groups fused with cycloalkyl groups. When fused with cycloalkyl groups, the aromatic group is the linking point. It usually contains 6 to 14 carbon atoms, and further contains 6 to 10 carbon atoms. Non-limiting examples include phenyl, naphthyl, anthracenyl, phenanthrenyl, The aryl group may be optionally substituted with a substituent.

[0125] "Heterocycloalkyl" refers to a saturated or partially unsaturated non-aromatic carbocyclic ring containing 1, 2, 3, or 4 heteroatoms selected from N, S, and O. Heterocycloalkyl can be monocyclic, bicyclic, or polycyclic. Bicyclic or polycyclic rings can be bridged, fused, spirocyclic, or a combination thereof. Bicyclic or polycyclic rings can include one or more aromatic or heteroaromatic rings, but the ring system as a whole is not aromatic. The attachment point can be on the aromatic ring or on the non-aromatic ring. Heterocycloalkyl groups are usually 3 to 20-membered rings. When they are monocyclic heterocycloalkyl groups, they are usually 3 to 15-membered rings, or 3-10-membered rings, or 3-8-membered rings, or 3-6-membered rings; when they are bicyclic or polycyclic heterocycloalkyl groups, they are usually 5-12-membered rings, or 5-11-membered rings, or 6-9-membered rings. The heteroatoms N and S include their oxidation states. Non-limiting examples of heterocycloalkyl groups include azetidinyl, morpholinyl, piperazinyl, piperidinyl, tetrahydropyranyl, oxetanyl, pyranyl, azolidinyl, azolinyl, oxolanyl, oxekinyl, and the like. Heterocycloalkyl groups may be optionally substituted with substituents.

[0126] "Heteroaromatic ring" or "heteroaryl" unless otherwise specified refers to a ring containing 1 to 4 heteroatoms selected from N, O or S and their oxidation states and having aromatic properties, which may be monocyclic, bicyclic or polycyclic, and the bicyclic or polycyclic rings may be bridged, parallel, spirocyclic or combinations thereof; when bicyclic or polycyclic, it may be a fusion of a heteroaryl and an aryl, or a fusion of a heteroaryl and a heteroaryl, wherein either the heteroaryl or the aryl may be a connection site. Non-limiting examples include furanyl, thienyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, pyridyl, pyrimidinyl, pyridazinyl, pyrazinyl, indolyl, purinyl, The heteroaryl group may be optionally substituted by a substituent.

[0127] "Substitution" or "substituent" unless otherwise specified refers to any substitution at a position permitted by chemical theory, and the number of substituents complies with the chemical bond rules. Exemplary substituents include but are not limited to: C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-8 Heteroalkyl, C 5-12 Aryl, 5-12 membered heteroaryl, hydroxyl, C 1-6Alkoxy, C 5-12 Aryloxy, thiol, C 1-6 Alkylthio, cyano, halogen, C 1-6 Alkylthiocarbonyl, C 1-6 Alkylcarbamoyl, N-carbamoyl, nitro, silyl, sulfinyl, sulfonyl, sulfoxide, halo 1-6 Alkyl, halogenated C 1-6 Alkoxy, amino, phosphonic acid, -CO2(C 1-6 alkyl), -OC(=O)(C 1-6 alkyl), -OCO2(C 1-6 alkyl), -C(=O)NH2, -C(=O)N(C 1- 6-alkyl)2,-OC(=O)NH(C 1-6 alkyl), -NHC(=O)(C 1-6 alkyl), -N(C 1-6 alkyl)C(=O)(C 1-6 Alkyl), -NHCO2(C 1-6 alkyl), -NHC(=O)N(C 1-6 alkyl)2,-HC(=O)NH(C 1-6 alkyl), -NHC(=O)NH2, -NHSO2(C 1-6 alkyl), -SO2N(C 1-6 Alkyl)2,-SO2NH(C 1-6 alkyl), -SO2NH2, -SO2C 1-6 Alkyl, etc.

[0128] "Optional" or "optionally" means that the subsequently described event or circumstance may but need not occur, and the description includes instances where the event or circumstance occurs and instances where it does not. For example, "alkyl optionally substituted with F" means that the alkyl group may but need not be substituted with F, and the description includes instances where the alkyl group is substituted with F and instances where the alkyl group is not substituted with F.

[0129] "Pharmaceutically acceptable salts" refer to salts of the compounds of the present invention that retain the biological effectiveness and properties of the free acids or free bases, and the free acids are reacted with non-toxic inorganic or organic bases, or the free bases are reacted with non-toxic inorganic or organic acids.

[0130] A "pharmaceutical composition" refers to a mixture of one or more compounds, stereoisomers, pharmaceutically acceptable salts, or cocrystals described herein, with other ingredients, wherein the other ingredients include physiologically / pharmaceutically acceptable carriers and / or excipients.

[0131] "Carrier" refers to a system that does not cause significant irritation to the organism and does not eliminate the biological activity and properties of the administered compound, and can change the way the drug enters the human body and its distribution in the body, control the release rate of the drug and deliver the drug to the target organ. Non-limiting examples include microcapsules and microspheres, nanoparticles, liposomes, etc.

[0132] An "excipient" is a substance that is not itself a therapeutic agent but serves as a diluent, adjuvant, binder, and / or vehicle that is added to a pharmaceutical composition to improve its handling or storage properties or to allow or facilitate the formation of a compound or pharmaceutical composition into a unit dosage form for administration. As known to those skilled in the art, excipients can serve a variety of functions and can be described as wetting agents, buffers, suspending agents, lubricants, emulsifiers, disintegrants, absorbents, preservatives, surfactants, colorants, flavoring agents, and sweeteners. Examples of excipients include, but are not limited to: (1) sugars such as lactose, glucose, and sucrose; (2) starches such as corn starch and potato starch; (3) cellulose and its derivatives such as sodium carboxymethylcellulose, ethylcellulose, cellulose acetate, hydroxypropyl methylcellulose, hydroxypropyl cellulose, microcrystalline cellulose, and cross-linked carboxymethylcellulose (e.g., cross-linked sodium carboxymethylcellulose); (4) tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients such as cocoa butter and suppository waxes; (9) oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, etc. and soybean oil; (10) glycols such as propylene glycol; (11) polyols such as glycerol, sorbitol, mannitol and polyethylene glycol; (12) esters such as ethyl oleate and ethyl laurate; (13) agar; (14) buffers such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19) ethanol; (20) pH buffer solutions; (21) polyesters, polycarbonates and / or polyanhydrides; and (22) other non-toxic compatible substances used in pharmaceutical preparations.

[0133] The compounds of the present invention may exist in specific geometric or stereoisomeric forms. All such compounds of the present invention, including cis and trans isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers, (D)-isomers, (L)-isomers, and racemic mixtures and other mixtures thereof, such as enantiomerically or diastereomerically enriched mixtures, all of which are within the scope of the present invention. Additional asymmetric carbon atoms may be present in the substituents of the compounds of the present invention. All of these isomers and their mixtures are within the scope of the present invention. In certain embodiments, preferred compounds are those isomeric compounds that exhibit superior biological activity. Purified or partially purified isomers and stereoisomers of the compounds of the present invention, or racemic mixtures or diastereomeric mixtures, are also within the scope of the present invention. Purification and separation of such substances can be achieved by standard techniques known in the art.

[0134] "Isomers" include "stereoisomers" and "tautomers." "Stereoisomers" refer to isomers in which the atoms or groups of atoms in a molecule have the same order of attachment but different spatial arrangements. Stereoisomers include cis-trans isomers, optical isomers, and conformational isomers. "Tautomers" refer to compounds that can be converted into each other through a reversible chemical reaction called tautomerization, usually caused by the concomitant migration of hydrogen atoms and π bonds (double or triple bonds). Examples include the following pairs of compounds: aldehyde / ketone-enol, imine-enamine.

[0135] "Solvate" refers to a substance formed by a compound of the present invention or a salt thereof and a stoichiometric or non-stoichiometric amount of a solvent bound to the compound or salt thereof by non-covalent forces between the molecules. When the solvent is water, the solvate is a hydrate.

[0136] A "cocrystal" is a crystal formed by the active pharmaceutical ingredient (API) and cocrystal former (CCF) bound together by hydrogen bonds or other non-covalent bonds. Both the API and CCF are solid in their pure form at room temperature, and the components exist in a fixed stoichiometric ratio. A cocrystal is a multi-component crystal, encompassing both binary cocrystals formed between two neutral solids and multi-component cocrystals formed between a neutral solid and a salt or solvate. DETAILED DESCRIPTION

[0137] The present invention will be described in detail below through examples. Where specific conditions are not specified in the examples, the experimental methods are carried out according to conventional conditions. The examples are provided to better illustrate the present invention, but it should not be understood that the present invention is limited to the examples. Any non-essential improvements and adjustments made by those skilled in the art to the embodiments based on the above invention are still within the scope of protection of the present invention.

[0138] Test Method

[0139] The structures of the compounds were determined by nuclear magnetic resonance (NMR) or / and mass spectrometry (MS). NMR shifts (δ) were expressed in 10 -6 The unit of (ppm) is given. NMR measurements were performed using a Bruker Avance III 400 and Bruker Avance 300 NMR spectrometer. The solvents used were deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), and deuterated methanol (CD3OD). The internal standard was tetramethylsilane (TMS).

[0140] MS was determined using (Agilent 6120B (ESI) and Agilent 6120B (APCI));

[0141] HPLC determination was performed using an Agilent 1260DAD high pressure liquid chromatograph (Zorbax SB-C 18 100 × 4.6 mm, 3.5 μM);

[0142] Thin layer chromatography silica gel plates used were Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plates. The specifications of the silica gel plates used for thin layer chromatography (TLC) were 0.15 mm to 0.20 mm, and the specifications used for thin layer chromatography separation and purification products were 0.4 mm to 0.5 mm.

[0143] Column chromatography generally uses Yantai Huanghai silica gel 200-300 mesh silica gel as the carrier.

[0144] Example 1

[0145] Step 1: Under nitrogen, lithium aluminum deuteride (1.96 g, 46.7 mmol) was dispersed in ultra-dry tetrahydrofuran (100 mL), cooled to -10°C, and compound 1A (10 g, 46.7 mmol, dissolved in 100 mL of ultra-dry tetrahydrofuran) was slowly added dropwise. After the addition was complete, the mixture was allowed to react at -10°C for 1 hour. After the reaction was complete, water (2 mL), 15% NaOH (2 mL), and water (6 mL) were slowly added dropwise to the reaction solution. The mixture was stirred at room temperature for 30 minutes, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain compound 1B (7.5 g crude product).

[0146] LC-MS (ESI): m / z = 171.0 [M-17] + .

[0147] Step 2: Under nitrogen, compound 1B (5.0 g, 26.6 mmol) and triphenylphosphine (10.5 g, 40.1 mmol) were dissolved in dichloromethane (100 mL). Carbon tetrabromide (13.3 g, 40.1 mmol) was added portionwise to the reaction mixture. After addition, the mixture was allowed to react at room temperature for 1 hour. The mixture was then concentrated under reduced pressure, and the resulting residue was separated by column chromatography (100% petroleum ether) to afford compound 1C (6.0 g, 90.2% yield).

[0148] Step 3: Under nitrogen, NaH (60% (w / w), 1.92 g, 48.0 mmol) was dispersed in ultra-dry tetrahydrofuran (70 mL). The temperature was lowered to 0°C, and diethyl malonate (9.22 g, 57.6 mmol) was slowly added dropwise. After completion, the mixture was stirred at room temperature for 1 h. After the temperature was lowered to 0°C, 1C (6.0 g, 24.0 mmol, dissolved in 30 mL of ultra-dry tetrahydrofuran) was slowly added dropwise. After completion, the mixture was allowed to react at room temperature overnight. After completion, the reaction mixture was poured into ice water and extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. This was then separated by column chromatography (petroleum ether:ethyl acetate (v:v) = 10:1) to afford compound 1D (2.6 g, 32.8%).

[0149] LC-MS (ESI): m / z = 331.0 [M+H] + .

[0150] Step 4: Under nitrogen, compound 1D (2.66 g, 8.06 mmol), NaCl (935 mg, 16.1 mmol), water (290 mg, 16.1 mmol), and DMSO (130 mL) were added to the reaction flask in this order. After addition, the temperature was raised to 180°C and the reaction was allowed to proceed for 3.5 hours. After completion of the reaction, the reaction solution was poured into water and extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product, which was separated by column chromatography (petroleum ether:ethyl acetate (v:v) = 20:1) to afford compound 1E (1.8 g, yield: 86.5%).

[0151] LC-MS (ESI): m / z = 259.1 [M+H] + .

[0152] Step 5: Compound 1E (1.8 g, 6.97 mmol), anhydrous methanol (50 mL), water (10 mL), and lithium hydroxide (502 mg, 20.9 mmol) were added to the reaction flask in sequence. After addition, the mixture was allowed to react at room temperature overnight. After completion of the reaction, the mixture was concentrated under reduced pressure, diluted with water (20 mL), and the pH of the system was adjusted to 3-4. The filter cake was collected by filtration, washed with water, and dried to obtain compound 1F (1.4 g crude product), which was used directly in the next reaction.

[0153] LC-MS (ESI): m / z = 230.9 [M+H] + .

[0154] Step 6: Under nitrogen, compound 1F (1.4 g, 6.08 mmol) and N,N-dimethylformamide (0.2 mL) were dissolved in ultra-dry dichloromethane (30 mL). The temperature was lowered to 0°C, and oxalyl chloride (2.3 g, 18.2 mmol) was slowly added dropwise. After addition, the mixture was allowed to react at room temperature for 1 hour. After completion of the reaction, the mixture was concentrated under reduced pressure, ultra-dry dichloromethane (40 mL) was added, the temperature was lowered to 0°C, and AlCl₃ (2.4 g, 18.2 mmol) was added portionwise. After addition, the mixture was allowed to react at room temperature for 2 hours. After completion of the reaction, the reaction mixture was poured into ice water and extracted with dichloromethane. The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was separated by column chromatography (petroleum ether:ethyl acetate (v:v) = 5:1) to afford compound 1G (0.7 g, yield: 54.3%).

[0155] LC-MS (ESI): m / z = 212.9 [M+H] + .

[0156] Step 7: Compound 1G (700 mg, 3.3 mmol), tri-n-butyltinmethanol (1.59 g, 4.95 mmol), and Xphos Pd G2 (0.26 g, 0.33 mmol) were added to a reaction flask, dissolved with 1,4-dioxane (40 mL), and the atmosphere was purged with nitrogen three times. The reaction was allowed to proceed at 90°C for 3 hours. After completion of the reaction, monitored by TLC, the mixture was cooled to room temperature and concentrated under reduced pressure. The resulting residue was purified by column chromatography (petroleum ether:ethyl acetate (v:v) = 1:1) to afford compound 1H (0.5 g, yield: 92.4%).

[0157] LC-MS (ESI): m / z = 165.2 [M+H] + .

[0158] Step 8: Under nitrogen, compound 1H (500 mg, 3.05 mmol) and triphenylphosphine (1.2 g, 4.57 mmol) were dissolved in dichloromethane (30 mL). CBr (1.51 g, 4.57 mmol) was added portionwise to the reaction mixture. After addition, the mixture was allowed to react at room temperature for 1 hour. Completion of the reaction was monitored by TLC. The residue was concentrated under reduced pressure and separated by column chromatography (petroleum ether:ethyl acetate (v:v) = 5:1) to afford compound 1I (650 mg, yield: 94.3%).

[0159] Step 9: Compound 1I (200 mg, 0.88 mmol), compound 1J (synthesized by the method described in reference patent WO2021231630 A1) (240 mg, 0.88 mmol) and cesium carbonate (431 mg, 1.33 mmol) were added to the reaction flask, dissolved with acetonitrile (10 mL), and then reacted at 50 ° C for 4 hours. After the completion of the reaction monitored by TLC, it was cooled to room temperature and concentrated under reduced pressure. The resulting residue was purified by column chromatography (dichloromethane: ethyl acetate (v:v) = 1:1) to obtain compound 1 (210 mg, yield: 57.0%).

[0160] 1 HNMR(400MHz,DMSO-d6)δ9.15(s,2H),8.13-8.11(m,1H),7.74-7.72(m,1H),7.6 2-7.56(m,2H),7.17-7.14(m,1H),5.40(s,2H),5.14-5.07(m,2H),2.61(s,2H);

[0161] LC-MS (ESI): m / z = 419.1 [M+H] + .

[0162] Example 2

[0163] Step 1: Dissolve 2,2-difluoroethanol-1-ol (16.5 g, 201.2 mmol) in tetrahydrofuran (50 mL) and cool to 0°C. Slowly add sodium hydride (7.44 g, 310 mmol) in portions. Return to room temperature and stir for 40 minutes. Then, add compound 2A (30 g, 155.1 mmol) dissolved in tetrahydrofuran (50 mL) and continue the reaction for 3 hours. After completion of the reaction, dilute with water and extract three times with ethyl acetate. The organic phases are combined, dried, and concentrated to obtain compound 2B (36 g crude product), which is used directly in the next step.

[0164] LC-MS (ESI): m / z = 239.1 [M+H] + .

[0165] Step 2: Dissolve 2B (38 g, 158.98 mmol), potassium acetate (39.01 g, 397.45 mmol), pinacol diboronate (52.48 g, 206.67 mmol), and XPhos Pd G2 (12.51 g, 15.90 mmol) in 1,4-dioxane (500 mL). The atmosphere was purged with nitrogen three times, and then stirred at 100°C for 3 hours. After the reaction was complete, the mixture was filtered while hot, and the filter cake was rinsed four times with ethyl acetate and dichloromethane. The filtrate was collected and concentrated to obtain compound 2C (32 g crude product), which was used directly in the next reaction.

[0166] LC-MS (ESI): m / z = 205.2 [M+H] + .

[0167] Step 3: Compound 2C (30 g, 147.10 mmol), 6-bromo-3-pyridazinone (30.71 g, 176.52 mmol), tripotassium phosphate (40.59 g, 191.23 mmol), and PEPPSI-SIPR catalyst (10.02 g, 14.71 mmol) were dissolved in a mixed solvent of 1,4-dioxane (360 mL) and water (120 mL). The atmosphere was then replaced with nitrogen three times and stirred at 90°C for 3 hours. After cooling, the mixture was filtered and the filter cake was rinsed with water, ethyl acetate, and dichloromethane. The filter cake was collected and dried to obtain compound 2D (27 g, yield: 72%).

[0168] LC-MS (ESI): m / z = 255.1 [M+H] + .

[0169] Step 4: Compound 1I (200 mg, 0.88 mmol), compound 2D (225 mg, 0.88 mmol), and cesium carbonate (431 mg, 1.33 mmol) were added to a reaction flask, dissolved in acetonitrile (10 mL), and reacted at 50°C for 4 hours. After completion of the reaction, monitored by TLC, the mixture was cooled to room temperature and concentrated under reduced pressure. The resulting residue was purified by column chromatography (DCM:EA = 1:1) to provide compound 2 (230 mg, yield: 65.3%).

[0170] 1 HNMR(400MHz,DMSO-d6)δ9.12(s,2H),8.12-8.09(m,1H),7.74-7.72(m,1H),7.61-7.56(m ,2H),7.16-7.14(m,1H),6.57-6.30(m,1H),5.40(s,2H),4.73-4.64(m,2H),2.61(s,2H);

[0171] LC-MS (ESI): m / z = 401.2 [M+H] + .

[0172] Example 3 and Example 4

[0173] Step 1: Add a solution of diethylzinc (102.5 mL, 102.5 mmol, 1 M) to dichloromethane (100 mL) under nitrogen protection and cool to 0°C. Then, add a solution of trifluoroacetic acid (7.6 mL, 102.5 mmol) in dichloromethane (10 mL) and react for half an hour. Then, add a solution of diiodomethane (8.3 mL, 102.5 mmol) in dichloromethane (10 mL) and react for half an hour. Then, add a solution of compound 3A (10 g, 51.4 mmol) in dichloromethane (10 mL) and slowly warm to room temperature to react for 2 hours. After the reaction is complete, the reaction solution is concentrated to dryness under reduced pressure, and the residue is purified by column chromatography (ethyl acetate: petroleum ether (v:v) = 0% to 5%) to obtain compound 3B (9.3 g, yield: 85%).

[0174] 1 H NMR(400MHz,DMSO-d6)δ7.32(s,1H),7.23(d,2H),3.16–3.08(m,1H),2.94–2.84(m, 1H),2.40–2.30(m,1H),1.93–1.82(m,1H),1.11–1.05(m,1H),-0.02–-0.09(m,1H).

[0175] Step 2: Compound 3B (3 g, 15.4 mmol), (tributyltin)methanol (14.8 g, 46.14 mmol), and XPhos Pd G2 (0.24 g, 0.31 mmol) were added sequentially to 1,4-dioxane (30 mL). The atmosphere was replaced with nitrogen three times, and the mixture was heated to 90°C and stirred for 3 hours. After completion of the reaction, the mixture was cooled to room temperature and concentrated to dryness under reduced pressure. The residue was purified by column chromatography (ethyl acetate:petroleum ether (v:v) = 0% to 10%) to obtain compound 3C (1.8 g, yield: 73%).

[0176] 1 H NMR(400MHz,DMSO-d6)δ7.21(d,1H),7.08(s,1H),7.03–6.96(m,1H),4.99(s,1H),4.40(s,2H),3.15–3.05(m,1H ),2.85(d,1H),2.36–2.30(m,1H),1.91–1.79(m,1H),1.09–1.01(m,1H),0.92–0.85(m,1H),-0.11–-0.19(m,1H).

[0177] Step 3: Compound 3C (1.5 g, 9.36 mmol) was dissolved in dichloromethane (15 mL), and carbon tetrabromide (3.41 g, 10.3 mmol) and triphenylphosphine (2.95 g, 11.23 mmol) were added. The mixture was stirred at room temperature for 4 hours. After completion of the reaction, the reaction solution was concentrated to dryness under reduced pressure, and the residue was purified by column chromatography (single mobile phase: petroleum ether) to obtain compound 3D (1.4 g, yield: 67%).

[0178] Step 4: Compound 2D (0.1 g, 0.62 mmol), compound 3D (0.13 g, 0.51 mmol), and cesium carbonate (0.33 g, 1.03 mmol) were dissolved in acetonitrile (2 mL) and stirred at 80°C for 2 hours. After cooling, the mixture was concentrated directly. The residue was added with water (5 mL) and extracted three times with ethyl acetate (5 mL x 3). The organic phases were combined. The organic phases were dried over anhydrous sodium sulfate and concentrated to dryness under reduced pressure. The resulting residue was purified by reverse-phase column chromatography (acetonitrile:water (v:v) = 45:55) to obtain compound 3I (12 mg, yield: 10%).

[0179] LC-MS (ESI): m / z = 397.1 [M+H] + .

[0180] Step 5: Compound 3I (12.0 mg) was subjected to chiral separation by SFC to afford P1 (SFC elution time: 12.65–13.40 min, designated as compound 3) and P2 (SFC elution time: 14.10–15.23 min, designated as compound 4). SFC separation method: The sample was dissolved in DMF and filtered through a 0.45 μm filter to prepare a sample solution; instrument: SFC Prep 150AP; column: Daicel IG (19 mm x 250 mm); mobile phase: A for CO2; B for ethanol; gradient: 35% B; flow rate: 40 mL / min; column temperature: 25°C; wavelength: 220 nm; sample preparation: acetonitrile solution. After separation, the product was concentrated by rotary evaporation at 35°C, and the solvent was then dried in a lyophilizer at -80°C to afford compounds 3 (5 mg) and 4 (5 mg).

[0181] Compound 3 (SFC resolution peak time: 12.65-13.40 min): 1H NMR(400MHz,DMSO-d6)δ9.11(s,2H),8.09(d,1H),7.25(d,1H),7.17(s,1H),7.12(d,2H),6.62–6.27(m,1H),5.24(s,2H),4.7 1-4.65(m,2H),3.11-3.07(m,1H),2.85(d,1H),2.35-3.31(m,1H),1.90–1.80(m,1H),1.06-1.01(m,1H),-0.08—-0.11(m,1H).

[0182] Compound 4 (SFC resolution peak time: 14.10-15.23 min): 1 H NMR(400MHz,DMSO-d6)δ9.11(s,2H),8.09(d,1H),7.25(d,1H),7.17(s,1H),7.12(d,2H),6.60–6.28(m,1H),5.24(s,2H),4.7 1-4.65(m,2H),3.11-3.08(m,1H),2.85(d,1H),2.34–2.31(m,1H),1.89–1.80(m,1H),1.06-1.01(m,1H),-0.08—-0.11(m,1H).

[0183] Example 5 and Example 6

[0184] Step 1: Dissolve 2-methyl-2,2-difluoroethanol (3 g, 31.0 mmol) in tetrahydrofuran (50 mL). Add sodium hydride (0.99 g, 41.3 mmol) portionwise at 0°C. Return to room temperature and stir for 40 minutes. Then, inject compound 2A (4 g, 20.67 mmol) in tetrahydrofuran (50 mL) and continue stirring for 3 hours. Dilute with water and extract three times with ethyl acetate. The organic phase is collected, dried, and concentrated to yield compound 5A (5 g crude product), which is used directly in the next step.

[0185] LC-MS (ESI): m / z = 253.2 [M+H] + .

[0186] Step 2: Dissolve compound 5A (5 g, 19.76 mmol), potassium acetate (4.85 g, 49.40 mmol), pinacol diboronate (6.52 g, 25.70 mmol), and XPhos Pd G2 (1.55 g, 1.98 mmol) in 1,4-dioxane (60 mL). The mixture was purged with nitrogen three times and heated to 100°C under a nitrogen atmosphere with stirring for 3 hours. The mixture was filtered while hot, and the filter cake was rinsed four times with ethyl acetate and four times with dichloromethane. The organic phases were combined and concentrated under reduced pressure to afford compound 5B (5 g crude product), which was used directly in the next reaction.

[0187] LC-MS (ESI): m / z = 219.3 [M+H] + .

[0188] Step 3: Compound 5B (5 g, 22.94 mmol), 6-bromo-3-pyridazinone (4.85 g, 27.53 mmol), tripotassium phosphate (6.33 g, 29.82 mmol), and PEPPSI-SIPR catalyst (1.56 g, 2.29 mmol) were dissolved in a mixture of 1,4-dioxane (60 mL) and water (20 mL). The atmosphere was replaced with nitrogen three times and heated to 90°C under a nitrogen atmosphere with stirring for 3 hours. After cooling, the mixture was filtered and the filter cake was rinsed with water, ethyl acetate, and dichloromethane. The filter cake was collected and dried to yield compound 5C (2.5 g crude product), which was used directly in the next reaction.

[0189] LC-MS (ESI): m / z = 269.0 [M+H] + .

[0190] Step 4: Compound 3D (0.15 g, 0.67 mmol), compound 5C (0.15 g, 0.56 mmol), and cesium carbonate (0.36 g, 1.12 mmol) were dissolved in acetonitrile (3 mL) and stirred at 80°C for 2 hours. After cooling, the mixture was concentrated. Water (5 mL) was added to the residue, which was extracted three times with ethyl acetate (5 mL x 3). The organic phases were collected, dried, and concentrated. The resulting residue was purified by reverse-phase column chromatography (acetonitrile:water (v:v) = 45:55) to afford compound 5D (120 mg, 52.2% yield).

[0191] LC-MS (ESI): m / z = 411.1 [M+H] + .

[0192] Step 5: Compound 5D (120.0 mg) was subjected to chiral separation to afford P1 (SFC elution time: 13.10–13.75 min, identified as compound 5) and P2 (SFC elution time: 14.15–14.80 min, identified as compound 6). SFC separation method: The sample was dissolved in DMF and filtered through a 0.45 μm filter to prepare a sample solution; instrument: SFC Prep 150AP; column: Daicel OJ (19 mm x 250 mm); mobile phase: A for CO2; B for ethanol (0.5% ammonia); gradient: 33% B; flow rate: 40 mL / min; column temperature: 25°C; wavelength: 220 nm; sample preparation: acetonitrile solution. After separation, the product was concentrated by rotary evaporation at 35°C, and the solvent was then dried in a lyophilizer at -80°C to afford compounds 5 (57 mg) and 6 (61 mg).

[0193] Compound 5 (SFC resolution peak time: 13.10-13.75 min): 1 H NMR(400MHz,DMSO-d6)δ9.11(s,2H),8.09(d,1H),7.24(d,1H),7.17(s,1H),7.12(d,2H),5.24(s,2H),4.72-4.65(m,2H),3.1 1-3.07(m,1H),2.85(d,1H),2.36-2.31(m,1H),1.88-1.82(m,1H),1.80-1.71(m,3H),1.06-1.01(m,1H),-0.08—-0.11(m,1H).

[0194] Compound 6 (SFC resolution peak time: 14.15-14.80 min): 1 H NMR(400MHz,DMSO-d6)δ9.11(s,2H),8.09(d,1H),7.25(d,1H),7.17(s,1H),7.12(d,2H),5.24(s,2H),4.72-4.65(m,2H),3.1 1-3.07(m,1H),2.85(d,1H),2.36-2.31(m,1H),1.88-1.82(m,1H),1.81-1.71(m,3H),1.06-1.01(m,1H),-0.08—-0.11(m,1H).

[0195] Example 7 and Example 8

[0196] Step 1: (R)-(-)-3-Hydroxytetrahydrofuran (Compound 7A) (2.05 g, 23.27 mmol) was added to tetrahydrofuran (50 mL) solvent, cooled to 0-10°C under nitrogen protection, and sodium hydride (0.45 g, 18.61 mmol) was slowly added. The mixture was stirred for half an hour, and 5-bromo-2-chloropyrimidine (3.00 g, 15.51 mmol) was slowly added. The reaction was allowed to react at room temperature for 2 hours. After completion of the reaction, the mixture was filtered, the filter cake was washed with ethyl acetate, and the organic phase was washed with saturated brine. The combined organic phases were dried over anhydrous sodium sulfate and concentrated under reduced pressure. The resulting residue was purified by column chromatography (ethyl acetate: petroleum ether (v:v) = 0-20%) to obtain Compound 7B (3.40 g, yield: 89.45%).

[0197] LC-MS (ESI): m / z = 245.0 [M+H] + .

[0198] Step 2: Compound 7B (3.40 g, 13.87 mmol) and pinacol diboron (3.52 g, 13.87 mmol) were added to 1,4-dioxane (50 mL) solvent. Potassium acetate (3.40 g, 34.67 mmol) was then added and the atmosphere was purged with nitrogen three times. XPhos Pd G2 (1.09 g, 1.39 mmol) was added and the atmosphere was purged with nitrogen three times. The temperature was raised to 100°C and the reaction was allowed to react for 2 hours. After completion of the reaction, the mixture was filtered and the filtrate was concentrated to dryness to obtain compound 7C (3.40 g crude product), which was used directly in the next reaction.

[0199] LC-MS (ESI): m / z = 211.1 [M+H] + .

[0200] Step 3: Compound 7C (3.40 g crude product), 6-bromo-3-pyridazinone (2.43 g, 13.87 mmol), tripotassium phosphate (2.94 g, 13.87 mmol), and PEPPSI-SIPR catalyst (0.94 g, 1.39 mmol) were dissolved in 1,4-dioxane (100 mL). The atmosphere was replaced with nitrogen three times and heated to 110°C under a nitrogen atmosphere with stirring for 3 hours. After cooling, the mixture was filtered and the filter cake was washed with ethyl acetate. The combined filtrates were washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated. The resulting residue was purified by column chromatography (ethyl acetate:petroleum ether (v:v) = 0-100%) to afford compound 7D (150 mg, yield: 4.16%).

[0201] LC-MS (ESI): m / z = 261.0 [M+H] + .

[0202] Step 4: Compound 7D (0.15 g, 0.58 mmol), compound 3D (0.13 g, 0.58 mmol), and cesium carbonate (0.38 g, 1.16 mmol) were dissolved in acetonitrile (20 mL) and stirred at 80°C for 2 hours. After cooling, the mixture was concentrated directly, and the residue was extracted with water and ethyl acetate. The organic phase was collected, dried over anhydrous sodium sulfate, and concentrated to dryness. The resulting residue was purified by reverse phase column chromatography (acetonitrile:water (v:v) = 5%-80%) to give compound 7E (71.00 mg, yield: 30.61%).

[0203] LC-MS (ESI): m / z = 403.1 [M+H] + .

[0204] Step 5: Compound 7E (71.00 mg) was chirally resolved to yield P1 (SFC elution time: 9.10–10.26 min, assumed to be compound 7) and P2 (SFC elution time: 13.13–13.93 min, assumed to be compound 8). Preparation method: Instrument: SFC Prep 150AP; Column: Daicel OJ-H (19 mm × 250 mm); Mobile phase: A for CO2; B for EtOH; Gradient: 35% B isocratic elution; Flow rate: 40 mL / min; Column temperature: 25°C; Wavelength: 220 nm; Cycle time: 25.0 min; Sample preparation: Sample concentration: 10 mg / mL, methanol solution; Injection: 10 mL per sample. After separation, the product was concentrated by rotary evaporation at 35°C, and the solvent was then dried in a lyophilizer at -80°C to yield Compound 7 (23 mg) and Compound 8 (23 mg).

[0205] Compound 7 (SFC resolution peak time: 9.10-10.26 min): 1 H NMR(400MHz, CDCl3)δ8.89(s,2H),7.54(d,1H),7.26–7.22(m,3H),7.04( d,1H),5.61-5.57(m,1H),5.32(s,2H),4.16-4.12(m,1H),4.10–3.91(m,3 H),3.17-3.11(m,1H),2.92-2.88(m,1H),2.34-2.20(m,3H),1.89–1.79(m ,1H),1.11-1.01(m,1H),0.04-0.01(m,1H); LC-MS(ESI):m / z=403.1[M+H] + .

[0206] Compound 8 (SFC resolution peak time: 13.13-13.93 min):1 H NMR(400MHz, CDCl3)δ8.89(s,2H),7.54(d,1H),7.26–7.21(m,3H),7.04( d,1H),5.60-5.59(m,1H),5.32(s,2H),4.16-4.12(m,1H),4.08–3.91(m,3 H),3.17-3.11(m,1H),2.92-2.88(m,1H),2.37–2.19(m,3H),1.87-1.80(m ,1H),1.06-1.01(m,1H),0.04-0.01(m,1H); LC-MS(ESI):m / z=403.1[M+H] + .

[0207] Example 9 and Example 10

[0208] Step 1: (S)-(+)-3-Hydroxytetrahydrofuran (Compound 9A) (2.05 g, 23.27 mmol) was added to tetrahydrofuran (50 mL) under nitrogen protection. The temperature was lowered to 0-10°C, and sodium hydride (0.45 g, 18.61 mmol) was slowly added. The mixture was stirred for half an hour, and 5-bromo-2-chloropyrimidine (3.00 g, 15.51 mmol) was slowly added. The reaction was allowed to react at room temperature for 2 hours. After the reaction was completed, the mixture was filtered, the filter cake was washed with ethyl acetate, and the organic phase was washed with saturated brine. The combined organic phases were dried over anhydrous sodium sulfate and concentrated to dryness under reduced pressure. The resulting residue was purified by column chromatography (ethyl acetate: petroleum ether (v:v) = 0-20%) to obtain Compound 9B (3.40 g, yield: 89.45%).

[0209] LC-MS(ESI): m / z=245.1, 247.1[M+H] + .

[0210] Step 2: Compound 9B (3.40 g, 13.87 mmol) and pinacol diboron (3.52 g, 13.87 mmol) were added to 1,4-dioxane (50 mL) solvent. Potassium acetate (3.40 g, 34.67 mmol) was then added and the atmosphere was purged with nitrogen three times. XPhos Pd G2 (1.09 g, 1.39 mmol) was added and the atmosphere was purged with nitrogen three times. The temperature was raised to 100°C and the reaction was allowed to react for 2 hours. After completion of the reaction, the mixture was filtered and the filtrate was concentrated to dryness to obtain compound 9C (4.5 g crude product), which was used directly in the next reaction.

[0211] LC-MS (ESI): m / z = 211.1 [M+H] + .

[0212] Step 3: Compound 9C (4.5 g crude product), 6-bromo-3-pyridazinone (2.43 g, 13.87 mmol), tripotassium phosphate (2.94 g, 13.87 mmol), and PEPPSI-SIPR catalyst (0.94 g, 1.39 mmol) were dissolved in 1,4-dioxane (100 mL). The atmosphere was replaced with nitrogen three times and heated to 110°C under a nitrogen atmosphere with stirring for 3 hours. After cooling, the mixture was filtered and the filter cake was washed with ethyl acetate. The organic phase was collected, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to dryness. The residue was purified by column chromatography (ethyl acetate:petroleum ether (v:v) = 0-100%) to obtain compound 9D (340 mg, yield: 9.42%).

[0213] LC-MS (ESI): m / z = 261.1 [M+H] + .

[0214] Step 4: Compound 9D (0.15 g, 0.58 mmol), compound 3D (0.13 g, 0.58 mmol) and cesium carbonate (0.38 g, 1.16 mmol) were dissolved in acetonitrile (20 mL), followed by stirring at 80°C for 2 hours. After cooling, the mixture was concentrated directly, and the residue was extracted with water and ethyl acetate. The organic phase was collected, dried over anhydrous sodium sulfate, and concentrated to dryness. The resulting residue was purified by reverse phase column chromatography (acetonitrile: water (v:v) = 5%-80%) to give compound 9E (101 mg, yield: 30.61%).

[0215] LC-MS (ESI): m / z = 403.1 [M+H] + .

[0216] Step 5: Compound 9E (101 mg) was chirally resolved to yield P1 (SFC elution time: 9.48–9.98 min, designated as compound 9) and P2 (SFC elution time: 13.83–14.73 min, designated as compound 10). Preparation method: Instrument: SFC Prep 150AP; Column: Daicel OJ-H (19 mm × 250 mm); Mobile phase: A for CO2; B for EtOH; Gradient: 35% B isocratic elution; Flow rate: 40 mL / min; Column temperature: 25°C; Wavelength: 220 nm; Cycle time: 25.0 min; Sample preparation: Sample concentration: 10 mg / mL, methanol solution; Injection: 10 mL per sample. After separation, the product was concentrated by rotary evaporation at 35°C, and the solvent was then dried in a lyophilizer at -80°C to yield compounds 9 (35 mg) and 10 (35 mg).

[0217] Compound 9 (SFC resolution peak time: 9.48-9.98 min):1 H NMR(400MHz, CDCl3)δ8.89(s,2H),7.54(d,1H),7.26–7.19(m,3H),7.04( d,1H),5.61-5.57(m,1H),5.32(s,2H),4.16-4.12(m,1H),4.08–3.89(m,3 H),3.17-3.11(m,1H),2.95–2.86(m,1H),2.36–2.20(m,3H),1.87-1.80(m ,1H),1.06-1.01(m,1H),0.04-0.01(m,1H); LC-MS(ESI):m / z=403.1[M+H] + .

[0218] Compound 10 (SFC resolution peak time: 13.83-14.73 min): 1 H NMR(400MHz, CDCl3)δ8.89(s,2H),7.54(d,1H),7.25–7.17(m,3H),7.04( d,1H),5.60-5.57(m,1H),5.32(s,2H),4.16-4.12(m,1H),4.09–3.90(m,3 H),3.17-3.11(m,1H),2.95–2.86(m,1H),2.34-2.29(m,3H),1.87-1.80(m ,1H),1.06-1.01(m,1H),0.04-0.01(m,1H); LC-MS(ESI):m / z=403.1[M+H] + .

[0219] Example 11, Example 12, Example 13 and Example 14

[0220] Step 1: Compound 11A (1.0 g, 8.14 mmol) was added to tetrahydrofuran (50 mL) and cooled to 0-10°C under nitrogen. Sodium hydride (0.33 g, 8.14 mmol) was slowly added, and the mixture was stirred for half an hour. 5-Bromo-2-chloropyrimidine (1.50 g, 7.75 mmol) was slowly added, and the mixture was allowed to react at room temperature for 2 hours. After completion of the reaction, the mixture was filtered, the filter cake was washed with ethyl acetate, and the organic phase was washed with saturated brine. The combined organic phases were dried over anhydrous sodium sulfate and concentrated to dryness under reduced pressure. The resulting residue was purified on a silica gel column (ethyl acetate:petroleum ether (v:v) = 0-20%) to obtain compound 11B (1.68 g, yield: 77.08%).

[0221] LC-MS(ESI): m / z=281.0, 283.0[M+H] +.

[0222] Step 2: Compound 11B (1.70 g, 6.05 mmol) and pinacol diboron (1.54 g, 6.05 mmol) were added to 1,4-dioxane (50 mL). Potassium acetate (1.48 g, 15.13 mmol) was then added and the atmosphere was purged with nitrogen three times. XPhos Pd G2 (0.48 g, 0.60 mmol) was added and the atmosphere was purged with nitrogen three times. The temperature was raised to 100°C and the reaction was allowed to react for 2 hours. After completion of the reaction, the mixture was filtered, and the filtrate was collected and concentrated to obtain compound 11C (1.5 g crude product), which was used directly in the next reaction.

[0223] LC-MS (ESI): m / z = 247.1 [M+H] + .

[0224] Step 3: Compound 11C (1.5 g crude product), 6-bromo-3-pyridazinone (1.16 g, 6.66 mmol), tripotassium phosphate (1.28 g, 6.05 mmol), and PEPPSI-SIPR catalyst (0.41 g, 0.60 mmol) were dissolved in 1,4-dioxane (50 mL). The solvent was replaced with nitrogen three times and heated to 110°C under a nitrogen atmosphere with stirring for 3 hours. After cooling, the mixture was filtered and the filter cake was washed with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to dryness. The residue was purified by column chromatography (ethyl acetate:petroleum ether (v:v) = 0-100%) to obtain compound 11D (150 mg, yield: 8.37%).

[0225] LC-MS (ESI): m / z = 297.1 [M+H] + .

[0226] Step 4: Compound 11D (0.13 g, 0.44 mmol), compound 3D (0.098 g, 0.44 mmol) and cesium carbonate (0.15 g, 1.10 mmol) were dissolved in acetonitrile (20 mL), followed by stirring at 80°C for 2 hours. After cooling, the mixture was directly concentrated. Water and ethyl acetate were added to the residue, the layers were separated and washed, and the organic phase was collected, dried over anhydrous sodium sulfate, and concentrated to dryness. The resulting residue was purified by reverse phase column chromatography (acetonitrile: water (v:v) = 5%-80%) to give compound 11E (96 mg, yield: 49.89%).

[0227] LC-MS (ESI): m / z = 439.3 [M+H] + .

[0228] Step 5: Compound 11E (96 mg) was separated by chiral SFC to obtain P1 (SFC separation peak time: 11.03-11.83 min, set as compound 11), P2 (SFC separation peak time: 13.02-14.01 min, set as compound 12), P3 (SFC separation peak time: 15.23-15.95 min, set as compound 13) and P4 (SFC separation peak time: 20.70-21.42 min, set as compound 14). Preparation method: Instrument: SFC Prep 150AP; Column: Daicel OJ-H (19 mm × 250 mm); Mobile phase: A for CO2; B for EtOH; Gradient: 35% B isocratic elution; Flow rate: 40 mL / min; Column temperature: 25°C; Wavelength: 220 nm; Cycle time: 25.0 min; Sample preparation: Sample concentration: 10 mg / mL, methanol solution; Injection: 10 mL per sample. After separation, the product was dried and concentrated by rotary evaporation at a bath temperature of 35°C, and the solvent was then dried in a lyophilizer at -80°C to obtain Compound 11 (15 mg), Compound 12 (12 mg), Compound 13 (15 mg), and Compound 14 (12 mg).

[0229] Compound 11 (SFC resolution peak time: 11.03-11.83 min): 1 H NMR(400MHz, CDCl3)δ8.93(s,2H),7.55(d,1H),7.25–7.19(m,3H),7.05( d,1H),5.67-5.61(m,1H),5.32(s,2H),4.48-4.44(m,1H),4.13-3.99(m,3 H),3.17-3.11(m,1H),2.93-2.88(m,1H),2.35-2.28(m,1H),1.87-1.80(m ,1H),1.06-0.88(m,1H),0.04-0.01(m,1H); LC-MS(ESI):m / z=439.3[M+H] + .

[0230] Compound 12 (SFC resolution peak time: 13.02-14.01 min): 1H NMR(400MHz, CDCl3)δ8.93(s,2H),7.55(d,1H),7.25–7.19(m,3H),7.05( d,1H),5.67-5.61(m,1H),5.32(s,2H),4.48-4.43(m,1H),4.15–3.98(m,3 H),3.17-3.11(m,1H),2.92-2.88(m,1H),2.35–2.27(m,1H),1.88–1.79(m ,1H),1.06-1.01(m,1H),0.04-0.01(m,1H); LC-MS(ESI):m / z=439.3[M+H] + .

[0231] Compound 13 (SFC resolution peak time: 15.23-15.95 min): 1 H NMR(400MHz, CDCl3)δ8.93(s,2H),7.55(d,1H),7.25–7.18(m,3H),7.05( d,1H),5.67-5.61(m,1H),5.32(s,2H),4.48-4.43(m,1H),4.15–3.97(m,3 H),3.17-3.11(m,1H),2.95–2.86(m,1H),2.35–2.27(m,1H),1.87-1.80(m ,1H),1.06-1.01(m,1H),0.04-0.01(m,1H); LC-MS(ESI):m / z=439.3[M+H] + .

[0232] Compound 14 (SFC resolution peak time: 20.70-21.42 min): 1 H NMR(400MHz, CDCl3)δ8.93(s,2H),7.55(d,1H),7.25–7.19(m,3H),7.05( d,1H),5.67-5.61(m,1H),5.32(s,2H),4.48-4.43(m,1H),4.15–3.98(m,3 H),3.17-3.11(m,1H),2.93-2.88(m,1H),2.36–2.26(m,1H),1.87-1.80(m ,1H),1.06-1.01(m,1H),0.04-0.01(m,1H); LC-MS(ESI):m / z=439.3[M+H] + .

[0233] Example 15

[0234] Step 1: Methyltriphenylphosphonium bromide (18.6 g, 52.12 mmol) was added to a reaction flask and dissolved in THF (120 mL). The atmosphere was purged with nitrogen three times and cooled to -20°C. Then, n-butyllithium (21 mL, 52.12 mmol, 2.5 M in hexanes) was added dropwise. After the addition was complete, the mixture was stirred at this temperature for 30 min. Compound 15A (10.0 g, 47.38 mmol) was then dissolved in THF (30 mL) and added dropwise to the above system. After the addition was complete, the mixture was slowly warmed to room temperature and allowed to react for 18 hours. After completion of the reaction, saturated ammonium chloride solution (50 mL) was added to quench the reaction and diluted with water (500 mL). The mixture was then extracted twice with ethyl acetate (100 mL x 2). The organic phases were combined, dried, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (PE:EA = 50:1) to afford compound 15B (2.2 g, yield: 22.21%).

[0235] 1 H NMR(400MHz,DMSO-d6)δ7.78-7.75(m,1H),7.41-7.36(m,1H),7.27-7.23(m,1H ),5.62-5.58(m,1H),5.10-5.06(m,1H),2.92-2.86(m,2H),2.78-2.71(m,2H).

[0236] Step 2: Compound 15B (2.2 g, 10.52 mmol) was dissolved in THF (30 mL) and cooled to 0°C. Borane tetrahydrofuran (15.78 mL, 15.78 mmol, 1.0 M solution in THF) was then added dropwise. After the addition was complete, the mixture was warmed to room temperature and reacted for 3 hours. The mixture was then cooled to 0°C, and 2 mol / L sodium hydroxide (27 mL, 53 mmol) and hydrogen peroxide (2.8 mL, 21.04 mmol, 30% wt) were added sequentially. The mixture was warmed to room temperature and reacted for 1 hour. After completion of the reaction, the mixture was diluted with water (100 mL) and extracted twice with ethyl acetate (50 mL x 2). The organic phases were combined, dried, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (PE:EA = 5:1) to obtain compound 15C (2.0 g, yield: 83.71%).

[0237] 1 H NMR(400MHz,DMSO-d6)δ7.47(s,1H),7.31-7.27(m,1H),7.18-7.13(m,1H),4.85-4.60(m,1H), 3.59-3.47(m,2H),3.24-3.16(m,1H),2.87-2.69(m,2H),2.18-2.08(m,1H),1.86-1.75(m,1H).

[0238] Step 3: Compound 15C (2.0 g, 8.81 mmol) was added to a reaction flask and dissolved in DCM (70 mL). Dess-Martin reagent (5.5 g, 13.21 mmol) was then added and allowed to react at room temperature for 1 hour. After completion of the reaction, as monitored by TLC, saturated sodium thiosulfate solution (100 mL) was added to quench the reaction. The product was then extracted twice with DCM (70 mL x 2). The organic phases were combined, dried, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (PE:EA = 10:1) to afford compound 15D (1.0 g, yield: 50.43%).

[0239] 1 H NMR(400MHz,DMSO-d6)δ9.74-9.20(m,1H),7.57(s,1H),7.43-7.39(m,1H), 7.26-7.22(m,1H),4.12-4.06(m,1H),2.89-2.82(m,2H),2.41–2.20(m,2H).

[0240] Step 4: Compound 15D (1.0 g, 4.44 mmol) was added to a reaction flask and dissolved in ethylene glycol (30 mL). Then, aqueous formaldehyde (10 mL) and 50% KOH solution (10 mL) were added and refluxed for 6 hours. After completion of the reaction, the mixture was cooled to room temperature, diluted with water (100 mL), and extracted three times with ethyl acetate (100 mL x 3). The organic phases were combined, dried, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (PE:EA = 1:1) to obtain compound 15E (0.85 g, yield: 74.46%).

[0241] 1 H NMR(400MHz,DMSO-d6)δ7.40-7.37(m,1H),7.31-7.26(m,1H),7.15-7.10(m,1H ),4.67-4.61(m,2H),3.51-3.46(m,4H),2.81-2.74(m,2H),1.95-1.88(m,2H).

[0242] Step 5: Compound 15E (0.85 g, 3.31 mmol) was added to a reaction flask and dissolved in THF (30 mL). Carbon tetrachloride (2.04 g, 13.24 mmol) was then added, cooled to -48°C, and a solution of tri(dimethylamino)phosphine (0.56 g, 3.64 mmol) in THF (10 mL) was added dropwise. After the addition was complete, the mixture was transferred to an ice bath and slowly warmed to room temperature for 3 hours. The mixture was then concentrated under reduced pressure, and the resulting residue was dissolved in methanol (50 mL). Sodium methoxide (2.24 g, 13.24 mmol, 30% wt) was then added and the mixture was reacted at 90°C for 1 hour. After the reaction was completed as monitored by TLC, the mixture was cooled to room temperature and concentrated under reduced pressure. Water (100 mL) was added to the residue, and the mixture was extracted twice with ethyl acetate (50 mL*2). The organic phases were combined and dried, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (PE:EA=10:1) to obtain compound 15F (0.40 g, yield: 50.54%).

[0243] 1 H NMR (400MHz, DMSO-d6) δ7.82-7.79(m,1H),7.41-7.37(m,1H),7.20-7.16(m,1H),4.71-4.65(m,4H),2.82-2.76(m,2H),2.43-2.38(m,2H).

[0244] Step 6: Compound 15F (0.36 g, 1.51 mmol), tri-n-butyltinmethanol (0.97 g, 2.26 mmol), and Xphos Pd G2 (0.12 g, 0.15 mmol) were added to a reaction flask, dissolved with 1,4-dioxane (15 mL), and the atmosphere was replaced with nitrogen three times. The reaction was continued at 90°C for 4 hours. After completion of the reaction as monitored by TLC, the mixture was cooled to room temperature and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (PE:EA = 1:1) to obtain compound 15G (0.23 g, yield: 80.07%).

[0245] 1 H NMR(400MHz,DMSO-d6)δ7.60(s,1H),7.18–7.11(m,2H),5.18-5.12(m,1H), 4.72-4.65(m,4H),4.55-4.51(m,2H),2.83-2.76(m,2H),2.42–2.36(m,2H).

[0246] Step 7: Compound 15G (0.20 g, 1.05 mmol) was added to a reaction flask and dissolved in DCM (4 mL). Carbon tetrabromide (520 mg, 1.58 mmol) and triphenylphosphine (410 mg, 1.58 mmol) were added sequentially. The mixture was allowed to react at room temperature for 10 min. After completion of the reaction, monitored by TLC, the product was purified by silica gel column chromatography (PE:EA = 10:1) to afford compound 15H (65 mg, yield: 24.46%).

[0247] 1 H NMR(400MHz,DMSO-d6)δ7.74-7.71(m,1H),7.32-7.28(m,1H),7.22-7.18(m ,1H),4.77(s,2H),4.72-4.65(m,4H),2.84-2.78(m,2H),2.43–2.38(m,2H).

[0248] Step 8: Compound 1J (70 mg, 0.26 mmol), compound 15H (63 mg, 0.25 mmol), and cesium carbonate (163 mg, 0.50 mmol) were added to a reaction flask, dissolved in acetonitrile (10 mL), and reacted at 80°C for 2 hours. After completion of the reaction, monitored by TLC, the mixture was cooled to room temperature and concentrated under reduced pressure. The resulting residue was purified by reverse-phase column chromatography to afford compound 15 (22 mg, yield: 19.04%).

[0249] 1 H NMR(400MHz,DMSO-d6)δ9.15(s,2H),8.13-8.09(m,1H),7.72(s,1H),7.27–7.23(m,1H),7.20-7.1 3(m,2H),5.37(s,2H),5.15-5.06(m,2H),4.70-4.62(m,4H),2.82-2.76(m,2H),2.41-2.36(m,2H);

[0250] LC-MS (ESI): m / z = 445.8 [M+H] + .

[0251] Example 16

[0252] Step 1: Add 3-methylenecyclobutanecarbonitrile (2.91 g, 31.21 mmol) to tetrahydrofuran (50 mL), replace the atmosphere with nitrogen, and cool to -20°C. Slowly add a 1.0 mmol / L sodium bis(trimethylsilyl)amide solution in tetrahydrofuran (34 mL) dropwise. After complete addition, stir at room temperature for 30 minutes. Slowly add compound 16A (5.00 g, 28.37 mmol), and slowly warm the mixture to room temperature for 2 hours. After completion, add saturated aqueous ammonium chloride and ethyl acetate, separate the layers, and wash. The combined organic phases are dried over anhydrous sodium sulfate and concentrated. The resulting residue is purified on a silica gel column to yield compound 16B (3.90 g, 55.11% yield).

[0253] LC-MS(ESI):m / z=249.1,251.1[M+H] + .

[0254] Step 2: Compound 16B (3.4 g, 13.65 mmol) was added to 1,4-dioxane (50 mL), followed by potassium hydroxide (2.30 g, 40.95 mmol) and water (10 mL). The temperature was raised to 110°C and the reaction was allowed to react for 12 hours. After completion of the reaction, ethyl acetate and water were added, the layers were separated, and the mixture was washed. The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The resulting residue was purified on a silica gel column to obtain compound 16C (1.00 g, yield: 71.89%).

[0255] LC-MS(ESI):m / z=224.0,226.1[M+H] + .

[0256] Step 3: Compound 16C (1.00 g, 4.46 mmol) and (tributyltin)methanol (4.32 g, 13.44 mmol) were added to 1,4-dioxane (50 mL). After nitrogen purge, chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II) (XPhos Pd G2) (0.35 g, 0.45 mmol) was added. The atmosphere was again purged with nitrogen and the temperature was raised to 110°C. After completion of the reaction, the reaction was filtered, ethyl acetate and saturated brine were added, and the layers were separated and washed with saturated brine. The organic phases were combined and dried over anhydrous sodium sulfate. The organic phase was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to obtain compound 16D (0.71 g, yield: 90.91%).

[0257] LC-MS(ESI):m / z=157.1[M-OH-H] + .

[0258] Step 4: Compound 16D (0.71 g, 4.07 mmol) was dissolved in dichloromethane (50 mL), and triphenylphosphine (1.28 g, 4.88 mmol) was added. Carbon tetrabromide (1.48 g, 4.48 mmol) was slowly added under ice-cooling. The reaction was allowed to react at room temperature for 4 hours. After completion of the reaction, the reaction solution was concentrated under reduced pressure, toluene was added, stirred for 1 hour, and then filtered. The filtrate was washed with saturated brine, and the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain compound 16E (0.6 g crude product), which was used directly in the next reaction.

[0259] Step 5: Compound 16E (0.17 g, 0.73 mmol) and compound 1J (0.20 g, 0.73 mmol) were added sequentially to acetonitrile, followed by potassium carbonate (0.25 g, 1.82 mmol). The reaction was heated to 80°C for 2 hours. After completion, ethyl acetate and saturated brine were added, the layers separated, and washed. The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by reverse-phase silica gel column chromatography to yield compound 16 (152.00 mg, yield: 48.17%).

[0260] 1H NMR (400MHz, CDCl3) δ8.94(s,2H),8.76-8.64(m,1H),7.83-7.74(m,1H),7.64-7.54(m,1H),7.24-7. 15(m,1H),7.12-7.05(m,1H),5.39(s,2H),4.96–4.77(m,4H),3.72-3.58(m,1H),3.18–2.94(m,4H);

[0261] LC-MS (ESI): m / z = 430.2 [M+H] + .

[0262] Example 17

[0263] Step 1: Add compound 17A (5.00 g, 27.31 mmol) and zinc copper reagent (5.28 g, 40.96 mmol) to ether (50 mL) solvent. Dilute trichloroacetyl chloride (9.93 g, 54.62 mmol) and phosphorus oxychloride (4.61 g, 30.04 mmol) with ether (25 mL). Slowly add the diluted reagent to the reaction solution. Heat to 40 ° C for two hours, then react at room temperature for 24 hours. After the reaction is completed, filter, wash the filter cake with ether, wash the organic phase with saturated ammonium chloride and saturated brine, dry the organic phase over anhydrous sodium sulfate, and concentrate under reduced pressure. The residue is separated and purified by silica gel column to obtain compound 17B (8.00 g, yield: 99.63%).

[0264] LC-MS (ESI): m / z = 295.0 [M+H] + .

[0265] Step 2: Compound 17B (8.00 g, 27.21 mmol) was added to acetic acid (10 mL). Zinc powder (8.90 g, 136.05 mmol) was slowly added with stirring, and the temperature was raised to 120°C for 4 hours. After completion of the reaction, ethyl acetate was added, the mixture was filtered, and the filtrate was washed with saturated brine and saturated sodium bicarbonate. The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain compound 17C (3.80 g, yield: 62.04%).

[0266] LC-MS(ESI):m / z=225.1,227.1[M+H] + .

[0267] Step 3: Compound 17C (2.80 g, 12.44 mmol) was added to tetrahydrofuran (20 mL). The atmosphere was replaced with nitrogen three times, and TEBBE (0.5 mol / L, 8 mL) was added. The reaction was allowed to react at room temperature for 12 hours. After completion, 20% aqueous sodium hydroxide solution (2 mL) was added, and the mixture was filtered. The filter cake was washed with ethyl acetate, and the filtrate was washed with saturated brine. The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain compound 17D (2.20 g, yield: 79.27%).

[0268] LC-MS(ESI):m / z=223.2,225.2[M+H] + .

[0269] Step 4: Compound 17D (1.00 g, 4.46 mmol) and (tributyltin)methanol (4.32 g, 13.44 mmol) were added to 1,4-dioxane (50 mL). After nitrogen purge, chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II) (XPhos Pd G2) (0.35 g, 0.45 mmol) was added and the atmosphere was purged with nitrogen again. The temperature was raised to 110°C and the reaction mixture was reacted for 4 hours. After completion of the reaction, the mixture was filtered, ethyl acetate and saturated brine were added, the layers were separated, and the mixture was washed with saturated brine. The organic phases were combined and dried over anhydrous sodium sulfate. The organic phase was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to obtain compound 17E (0.71 g, yield: 90.91%).

[0270] LC-MS (ESI): m / z = 157.1 [M-OH] + .

[0271] Step 5: Compound 17E (0.71 g, 4.07 mmol) was added to dichloromethane, followed by triphenylphosphine (1.28 g, 4.88 mmol). Carbon tetrabromide (1.48 g, 4.48 mmol) was slowly added with stirring. The reaction was allowed to react at room temperature for 4 hours. After completion of the reaction, the reaction solution was concentrated to dryness under reduced pressure. Toluene was added, stirred for 1 hour, and then filtered. The filtrate was washed with saturated brine. The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain compound 17F (0.60 g crude product), which was used directly in the next reaction.

[0272] Step 6: Compound 17F (200 mg, 0.73 mmol), compound 1J (0.19 g, 0.80 mmol), and potassium carbonate (0.25 g, 1.82 mmol) were added sequentially to acetonitrile (20 mL) and heated to 80°C for 2 hours. After completion of the reaction, ethyl acetate and saturated brine were added, the layers separated, and washed. The organic phase was dried over anhydrous sodium sulfate and concentrated. The residue was purified by reverse-phase silica gel column chromatography to obtain compound 17 (157 mg, yield: 49.87%).

[0273] 1 H NMR(400MHz, CDCl3)δ8.94(s,2H),7.59(d,1H),7.48–7.38(m,2H),7.28–7.20(m,2H),7.09(d,1H),5.38 (s,2H),4.91-4.85(m,2H),4.83-4.81(m,2H),3.54-3.46(m,1H),3.12-3.04(m,2H),2.86–2.75(m,2H);

[0274] LC-MS (ESI): m / z = 429.2 [M+H] + .

[0275] Example 18

[0276] Step 1: Dissolve 1,1-dideutero-2,2,2-trifluoroethanol (3.96 g, 38.80 mmol) in tetrahydrofuran (50 mL). Cool the mixture to 0°C and slowly add sodium hydride (1.24 g, 31.04 mmol) in portions. Return the mixture to room temperature and stir for 40 minutes. Then, dissolve compound 2A (5 g, 25.87 mmol) in tetrahydrofuran (50 mL) and add the mixture. Continue the reaction for 3 hours. After completion, dilute with water and extract three times with ethyl acetate. The organic phases are combined, dried, and concentrated to yield compound 18B (6.1 g crude product), which is used directly in the next step.

[0277] LC-MS (ESI): m / z = 258.9 [M+H] +.

[0278] Step 2: Compound 18B (6 g, 23.16 mmol), potassium acetate (5.68 g, 57.9 mmol), pinacol diboronate (7.06 g, 27.79 mmol), and chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II) (XPhos Pd G2) (0.9 g, 1.16 mmol) were dissolved in 1,4-dioxane (100 mL). The atmosphere was exchanged with nitrogen three times and then stirred at 100°C for 3 hours. After completion of the reaction, the mixture was filtered while hot, and the filter cake was rinsed four times with ethyl acetate and dichloromethane. The filtrate was collected and concentrated to give compound 18C (10 g crude product), which was used directly in the next reaction.

[0279] LC-MS (ESI): m / z = 225.0 [M+H] + .

[0280] Step 3: Compound 18C (10 g, 44.65 mmol), 6-bromo-3-pyridazinol (5.47 g, 31.25 mmol), potassium carbonate (18.51 g, 133.9 mmol), and [1,3-bis(2,6-diisopropylphenyl)imidazol-2-ylidene](3-chloropyridyl)palladium(II) dichloride (PEPPS-IPR catalyst) (3.04 g, 4.46 mmol) were dissolved in a mixed solvent of 1,4-dioxane (100 mL) and water (20 mL). The mixture was then purged with nitrogen three times and stirred at 90°C for 3 hours. After completion of the reaction, the mixture was filtered, and the filter cake was washed with water, ethyl acetate, and dichloromethane, respectively. The filter cake was collected and dried to obtain compound 18D (6 g crude product), which was used directly in the next reaction.

[0281] LC-MS (ESI): m / z = 275.0 [M+H] + .

[0282] Step 4: Compound 1I (2.3 g, 10.21 mmol), compound 18D (3.5 g, 12.76 mmol), and potassium carbonate (5.3 g, 38.28 mmol) were added to a reaction flask, dissolved in DMF (50 mL), and reacted at 70°C for 4 hours. After completion of the reaction, monitored by TLC, the mixture was cooled to room temperature and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain compound 18 (1.5 g, yield: 27.47%).

[0283] 1H NMR (400MHz, DMSO-d6) δ9.15(s,2H),8.13(d,1H),7.75-7.72(m,1H),7.62(s,1H),7.57(d,1H),7.16(d,1H),5.40(s,2H),2.61(s,2H);

[0284] LC-MS (ESI): m / z = 421.1 [M+H] + .

[0285] Example 19

[0286] Step 1: Dissolve compound 19A (0.2 g, 1.23 mmol) in dichloromethane (10 mL), add thionyl chloride (0.16 g, 1.35 mmol), and stir at room temperature for 2 hours. After completion of the reaction, concentrate under reduced pressure to obtain compound 19B (0.24 g crude product), which is used directly in the next reaction.

[0287] Step 2: Methyltriphenylphosphonium bromide (1.09 g, 3.05 mmol) was added to tetrahydrofuran (20 mL). After nitrogen replacement, the temperature was lowered to -15°C, and n-butyllithium tetrahydrofuran solution (1.66 mL, 2 mol / L) was slowly added dropwise. The mixture was stirred and maintained at this temperature for 30 minutes. Compound 19B (0.50 g, 2.77 mmol) was added under nitrogen protection, and the temperature was slowly raised to room temperature for 1 hour. After completion of the reaction, toluene and saturated aqueous ammonium chloride were added, and the layers were separated and washed. The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain Compound 19C (180 mg, yield: 36.37%).

[0288] Step 3: Compound 19C (0.16 g, 0.90 mmol), compound 1J (0.20 g, 0.73 mmol), and potassium carbonate (0.20 g, 1.46 mmol) were added sequentially to acetonitrile (20 mL) and the reaction temperature was raised to 80°C for 2 hours. After completion of the reaction, ethyl acetate and saturated brine were added, the layers separated, and washed. The organic phase was concentrated under reduced pressure, and the residue was purified by reverse-phase silica gel column chromatography to obtain compound 19 (75.00 mg, yield: 24.63%).

[0289] 1H NMR(400MHz, CDCl3)δ8.93(s,2H),7.63–7.54(m,2H),7.38-7.31(m,1H),7.25-7.20(m,1H),7.09-7.04(m,1H),5 .48-5.44(m,1H),5.41-5.36(m,2H),5.07-5.00(m,1H),4.93-4.82(m,2H),2.99-2.89(m,2H),2.84-2.74(m,2H);

[0290] LC-MS (ESI): m / z = 415.4 [M+H] + .

[0291] Example 20

[0292] Step 1: Compound 20A (2.0 g, 8.36 mmol), (tributyltin)methanol (2.68 g, 8.36 mmol), and XPhos Pd G2 (0.66 g, 0.84 mmol) were added sequentially to 1,4-dioxane (30 mL). The atmosphere was purged with nitrogen three times, and the mixture was heated to 90°C with stirring for 3 hours. After completion of the reaction, the mixture was cooled to room temperature and concentrated to dryness under reduced pressure. The residue was purified by column chromatography to afford compound 20B (1.3 g, yield: 81.70%).

[0293] LC-MS (ESI): m / z = 191.1 [M+H] + .

[0294] Step 2: Dissolve compound 20B (1.0 g, 5.26 mmol) in dichloromethane (15 mL) and add phosphorus tribromide (0.48 g, 1.79 mmol). Stir at room temperature for 1 hour. After completion of the reaction, the reaction solution was concentrated to dryness under reduced pressure. The residue was purified by column chromatography to obtain compound 20C (0.92 g, yield: 69.14%).

[0295] LC-MS (ESI): m / z = 253.0 [M+H] + .

[0296] Step 3: Compound 1J (0.2 g, 0.73 mmol), compound 20C (0.18 g, 0.73 mmol), and potassium carbonate (0.3 g, 2.19 mmol) were dissolved in DMF (2 mL) and stirred at 70°C for 2 hours. After cooling, the mixture was concentrated directly. The residue was added with water (5 mL) and extracted three times with ethyl acetate (5 mL x 3). The organic phases were combined. The organic phases were dried over anhydrous sodium sulfate and concentrated to dryness under reduced pressure. The resulting residue was purified by reverse-phase column chromatography to obtain compound 20 (0.21 g, yield: 64.31%).

[0297] 1 H NMR(400MHz,DMSO-d6)δ9.16(s,2H),8.13(d,1H),7.77-7.75(m,1H),7.64(s,1H),7 .54(d,1H),7.16(d,1H),5.40(s,2H),5.14-5.08(m,2H),2.97(s,2H),1.12(s,6H);

[0298] LC-MS (ESI): m / z = 445.1 [M+H] + .

[0299] Example 21

[0300] Step 1: Compound 21A (1.8 g, 7.59 mmol), (tributyltin)methanol (2.44 g, 7.59 mmol), and XPhos Pd G2 (0.6 g, 0.76 mmol) were added sequentially to 1,4-dioxane (30 mL). The atmosphere was purged with nitrogen three times, and the mixture was heated to 90°C with stirring for 3 hours. After completion of the reaction, the mixture was cooled to room temperature and concentrated to dryness under reduced pressure. The residue was purified by column chromatography to afford compound 21B (1.3 g, 90.97% yield).

[0301] LC-MS (ESI): m / z = 189.1 [M+H] + .

[0302] Step 2: Dissolve compound 21B (1.0 g, 5.31 mmol) in dichloromethane (15 mL) and add phosphorus tribromide (0.49 g, 1.81 mmol). Stir at room temperature for 1 hour. After completion of the reaction, the reaction solution was concentrated to dryness under reduced pressure. The residue was purified by column chromatography to obtain compound 21C (1.1 g, yield: 82.45%).

[0303] LC-MS (ESI): m / z = 250.9 [M+H] + .

[0304] Step 3: 1J (100 mg, 0.37 mmol), compound 21C (93 mg, 0.37 mmol), and potassium carbonate (0.15 g, 1.11 mmol) were dissolved in DMF (2 mL) and stirred at 70°C for 2 hours. After cooling, the mixture was concentrated directly. The residue was added with water (5 mL) and extracted three times with ethyl acetate (5 mL x 3). The organic phases were combined. The organic phases were dried over anhydrous sodium sulfate and concentrated to dryness under reduced pressure. The resulting residue was purified by reverse-phase column chromatography to obtain compound 21 (50 mg, yield: 30.76%).

[0305] 1 H NMR(400MHz,DMSO-d6)δ9.15(s,2H),8.13(d,1H),7.76-7.74(m,1H),7.65(s,1H),7.6 0(d,1H),7.16(d,1H),5.42(s,2H),5.14-5.08(m,2H),3.21(s,2H),1.23-1.20(m,4H);

[0306] LC-MS (ESI): m / z = 443.2 [M+H] + .

[0307] Biological testing

[0308] 1. Rabbit skeletal muscle myosin activity analysis method

[0309] The purpose of this test is to evaluate the ability of compounds to inhibit the hydrolysis of ATP by rabbit skeletal muscle myosin. The assay is as follows:

[0310] 1. Solution preparation:

[0311] Preformed F-actin filaments (#AKF99-B, Cytoskeleton) were diluted to 1 mg / mL in reaction buffer (25 mM Tris-HCl, pH 7.5 + 35 mM KCl + 0.1 mM EGTA + 1 mM MgCl2);

[0312] Rabbit myosin (#MY02-A, Cytoskeleton) was diluted to 0.1 mg / mL in resuspension buffer (15 mM Tris-HCl, pH 7.5 + 0.2 M KCl + 1 mM MgCl2).

[0313] ATP (#BSA04-001, Cytoskeleton) was diluted to 3 mM working solution with 15 mM Tris-HCl (pH = 7.5);

[0314] 2. Experimental steps:

[0315] Add 13 μL reaction buffer + 12 μL F-actin to the actin control wells;

[0316] To the positive control wells, add 10 μL reaction buffer, 12 μL F-actin, and 3 μL myosin II.

[0317] Add 10 μL compound working solution + 12 μL F-actin + 3 μL myosin II to the compound wells;

[0318] Add 5 μL of ATP working solution to all test wells to start the reaction. Simultaneously add the Pi standard wells according to the manufacturer's instructions (#BK054, Cytoskeleton).

[0319] Mix gently and incubate at 37°C for 60 minutes.

[0320] · Add 70 μL of CytoPhos Reagent (#BK054, Cytoskeleton) to each well and incubate for 10 min to terminate the reaction.

[0321] 3. Detection:

[0322] Detect the OD value of each well using the endpoint method at 650 nm. Fit a standard curve based on Pi concentration and OD value to determine the Pi concentration in each test well. Inhibition (%) = (Pi positive control well - Pi test well) / Pi positive control well * 100%. Fit the Inhibition%-Enzyme concentration curve using Graphpad Prism 8 software to calculate the IC 50 .

[0323] 4. Test results: IC inhibitory activity of the compound on ATP hydrolysis of rabbit skeletal muscle myosin 50 Less than 1000 μM. Specific results of some examples are shown in Table 1.

[0324] Table 1 Inhibitory activity of compounds on ATP hydrolysis in rabbit skeletal muscle myosin

[0325] A represents 2.5 μM <IC 50 ≤20μM.

[0326] Conclusion: The compounds of the present invention, such as the compounds in the examples, have strong inhibitory effects on the hydrolysis of ATP by rabbit skeletal muscle myosin. The IC values ​​of some compounds are 50 Less than 1μM.

[0327] 2. Analysis of cardiac myosin SII activity

[0328] The purpose of this test is to evaluate the ability of compounds to inhibit the hydrolysis of ATP by cardiac myosin SII. The assay is as follows:

[0329] 1. Solution preparation:

[0330] Preformed F-actin filaments (#AKF99-B, Cytoskeleton) were diluted to 1 mg / mL in reaction buffer (15 mM Tris-HCl, pH 7.5 + 10 mM KCl + 0.1 mM EGTA + 2 mM MgCl2);

[0331] Cardiac muscle myosin (#MY03-A, Cytoskeleton) was diluted to 1 mg / mL in resuspension buffer (15 mM Tris-HCl, pH 7.5 + 0.2 M KCl + 1 mM MgCl2);

[0332] ATP (#BSA04-001, Cytoskeleton) was diluted to 3 mM working solution with 15 mM Tris-HCl (pH = 7.5);

[0333] 2. Preparation of compound working solution: the final concentration of the compound is 100 μM

[0334] 3. Experimental steps:

[0335] Add 10 μL compound working solution + 12 μL preformed F-actin filaments + 3 μL cardiac muscle myosin to the compound wells;

[0336] To the positive control wells, add 10 μL of reaction buffer containing DMSO, 12 μL of preformed F-actin filaments, and 3 μL of cardiac muscle myosin.

[0337] To the negative control wells, add 10 μL of reaction buffer containing DMSO, 12 μL of preformed F-actin filaments, and 3 μL of resuspension buffer.

[0338] Add 5 μL of 3 mM ATP working solution to all test wells to start the reaction, mix gently, and incubate at 37°C for 2 hours;

[0339] After the reaction, take 6 μL of the reaction solution, add 24 μL of reaction buffer and dilute it 5-fold. Then use a phosphate reagent kit (#BK054, Cytoskeleton) to detect the content of the product Pi.

[0340] ·Add sample to the Pi standard well according to the instructions;

[0341] 70 μL of CytoPhos Reagent (#BK054, Cytoskeleton) was added to each well and incubated at room temperature for 10 min before detection.

[0342] 4. Detection:

[0343] Detect the OD value of each well at 650 nm. Fit a standard curve based on the Pi standard concentration and OD value to determine the Pi concentration in each test well. Inhibition (%) = (positive control well - test well) / positive control well * 100%.

[0344] Table 2 Inhibition rate of compounds on cardiac myosin SII

[0345] Conclusion: The compounds of the present invention, such as the compounds in the examples, have no inhibitory activity on cardiac myosin SII.

[0346] 3. Smooth Muscle Myosin S1 Fragment Activity Analysis Method

[0347] The purpose of this test is to evaluate the ability of compounds to inhibit the hydrolysis of ATP by the smooth muscle myosin S1 fragment. The assay is as follows:

[0348] 1. Solution Preparation

[0349] F-actin polymer: Dissolve Cardiac Actin powder (#CS-AD99, Cytoskeleton) in buffer (5 mM Pipes-KOH solution, pH 7.0, containing 100 μM ATP and 500 μM DTT) to a 2 mg / mL solution and incubate at room temperature for 30 minutes. Then, add 2 mM MgCl2 and 2 mM EGTA and incubate at room temperature for 40 minutes.

[0350] Smooth muscle S1 fragment (#CS-MYS05, Cytoskeleton) was diluted to 0.7 mg / mL in pre-chilled PM12 buffer (12 mM Pipes-KOH solution, pH 7.0, containing 2 mM MgCl2) containing 1 mM DTT.

[0351] ATP (#BSA04-001, Cytoskeleton) was diluted to 5 mM working solution with 15 mM Tris-HCl (pH = 7.5);

[0352] 2. Compound working solution: Final concentration of compound is 100 μM

[0353] 3. Experimental steps:

[0354] Add 10 μL compound working solution + 6 μL F-actin polymer + 4 μL smooth muscle S1 fragment solution to the compound wells;

[0355] To the positive control wells, add 10 μL of PM12 buffer containing DMSO, 6 μL of F-actin polymer, and 4 μL of Smooth muscle S1 fragment solution.

[0356] To the negative control wells, add 10 μL of PM12 buffer containing DMSO, 6 μL of F-actin polymer, and 4 μL of PM12 buffer.

[0357] Add 10 μL of 5 mM ATP working solution to all test wells to start the reaction, mix gently, and incubate at 37°C for 20 minutes;

[0358] After the reaction, take 6 μL of the reaction solution, add 24 μL of PM12 buffer and dilute it 5-fold. Then use a phosphate reagent kit (#BK054, Cytoskeleton) to detect the content of the product Pi.

[0359] Standard samples were added according to the instructions, and 70 μL CytoPhos Reagent (#BK054, Cytoskeleton) was added to all wells and incubated at room temperature for 10 minutes before detection.

[0360] 4. Detection:

[0361] Detect the OD value of each well at 650nm. Fit the standard curve based on the Pi standard concentration and OD value to calculate the Pi concentration in each test well.

[0362] Inhibition (%) = (positive control well - test well) / positive control well * 100%.

[0363] Table 3 Inhibition rate of compounds on smooth muscle myosin S1

[0364] Conclusion: The compounds of the present invention, such as the compounds in the examples, have no inhibitory activity against smooth muscle myosin S1.

[0365] 4. Pharmacokinetics test in mice

[0366] 1. Experimental Animals: Male C57 mice, 22-25 g, 9 mice per compound, purchased from Beijing Huafukang Biotechnology Co., Ltd.

[0367] 2. Experimental Design: On the day of the experiment, C57 mice were randomly divided into groups according to body weight. They were fasted (but not watered) for 12–14 hours prior to dosing and fed 4 hours after dosing. Dosing was performed according to Table 4.1.

[0368] Table 4.1 Dosage Information

[0369] Note: Intravenous administration solvent: 10% DMA + 10% Solutol + 80% Saline; Oral administration solvent: 5% DMA + 5% Solutol + 90% Saline

[0370] (DMA: dimethylacetamide; Solutol: polyethylene glycol-15-hydroxystearate; Saline: normal saline)

[0371] Before and after drug administration, 0.06 mL of blood was collected from the eye socket under isoflurane anesthesia. The blood was placed in an EDTAK2 centrifuge tube and centrifuged at 5000 rpm at 4°C for 10 minutes to collect plasma. All samples were stored at -80°C prior to analysis and quantitative analysis was performed using LC-MS / MS.

[0372] Table 4. Pharmacokinetic parameters of test compounds in mouse plasma

[0373] *: The vehicle for the iv administration of compound 19 was 5% DMA + 5% Solutol + 90% Saline.

[0374] Conclusion: The compounds of the present invention, such as the compounds in the examples, have good pharmacokinetic characteristics in mice.

[0375] 5. Pharmacokinetics test in rats

[0376] 1. Experimental Animals: Male SD rats, approximately 220 g, 6 to 8 weeks old, 6 rats per compound, purchased from Chengdu Dashuo Experimental Animal Co., Ltd.

[0377] 2. Experimental Design: On the day of the experiment, SD rats were randomly divided into groups according to body weight. They were fasted (but not water) for 12–14 hours prior to dosing and fed 4 hours after dosing. Dosing was performed according to Table 5.1.

[0378] Table 5.1 Dosage Information

[0379] Note: Intravenous administration solvent: 5% DMA + 5% Solutol + 90% Saline; Oral administration solvent: 0.5% MC

[0380] (MC: methylcellulose)

[0381] Before and after drug administration, 0.15 mL of blood was collected from the eye socket under isoflurane anesthesia. The blood was placed in an EDTAK2 centrifuge tube and centrifuged at 5000 rpm at 4°C for 10 minutes to collect plasma. All samples were stored at -80°C prior to analysis and quantitative analysis was performed using LC-MS / MS.

[0382] Conclusion: The compounds of the present invention, such as the compounds in the examples, have good pharmacokinetic characteristics in rats.

[0383] 6. Pharmacokinetic Testing in Beagle Dogs

[0384] 1. Experimental Animals: Male beagle dogs, weighing approximately 8-11 kg, 6 per compound, purchased from Beijing Masi Biotechnology Co., Ltd.

[0385] 2. Experimental Method: On the day of the experiment, beagle dogs were randomly divided into groups according to body weight. They were fasted but not watered for 12-14 hours before administration and fed 4 hours after administration.

[0386] Before and after dosing, 1 mL of blood was collected from the jugular vein or limb vein and placed in an EDTAK2 centrifuge tube. Plasma was collected by centrifugation at 5000 rpm and 4°C for 10 minutes. All samples were stored at -80°C prior to analysis and quantitative analysis by LC-MS / MS.

[0387] Conclusion: The compounds of the present invention, such as the compounds in the examples, have good pharmacokinetic characteristics in beagle dogs.

[0388] 7. Monkey Pharmacokinetics Test

[0389] 1. Experimental Animals: Male cynomolgus monkeys, 3-5 kg, 3-6 years old, 4 per compound. Purchased from Suzhou Xishan Biotechnology Co., Ltd.

[0390] 2. Experimental Method: On the day of the experiment, monkeys were randomly divided into groups according to body weight. They were fasted but not watered for 14-18 hours before administration and fed 4 hours after administration.

[0391] Before and after dosing, 1.0 mL of blood was collected from a limb vein and placed in an EDTAK2 centrifuge tube. Plasma was collected by centrifugation at 5000 rpm and 4°C for 10 minutes. All samples were stored at -80°C prior to analysis and quantitative analysis was performed using LC-MS / MS.

[0392] Conclusion: The compounds of the present invention, such as the compounds in the examples, have good pharmacokinetic characteristics in monkeys.

[0393] 8. hERG potassium channel function test

[0394] 1. Experimental platform: electrophysiological manual patch clamp system

[0395] 2. Cell line: Chinese hamster ovary (CHO) cell line stably expressing hERG potassium channel

[0396] 3. Experimental method: CHO (Chinese Hamster Ovary) cells stably expressing hERG potassium channel were used to record hERG potassium channel current using whole-cell patch clamp technique at room temperature. Glass microelectrodes were drawn from glass electrode blanks (BF150-86-10, Sutter) using a drawing instrument. The tip resistance after perfusing the electrode liquid was about 2-5MΩ. The glass microelectrode was inserted into the amplifier probe to connect to the patch clamp amplifier. The clamping voltage and data recording were controlled and recorded by computer using pClamp 10 software, with a sampling frequency of 10kHz and a filter frequency of 2kHz. After obtaining the whole-cell recording, the cell was clamped at -80mV to induce hERG potassium current (I hERG ) was administered with a 2-second depolarization step from -80 mV to +20 mV, followed by repolarization to -50 mV for 1 second before returning to -80 mV. This voltage stimulus was administered every 10 seconds, and administration began after confirming that the hERG potassium current was stable (for at least 1 minute). Compounds were administered for at least 1 minute at each test concentration, and at least two cells (n ≥ 2) were tested for each concentration.

[0397] 4. Data processing: Data analysis was performed using pClamp 10, GraphPad Prism 5, and Excel. The degree of inhibition of hERG potassium current (peak hERG tail current induced at -50 mV) by different compound concentrations was calculated using the following formula:

[0398] Inhibition%=[1–(I / Io)]×100%

[0399] Wherein, Inhibition% represents the inhibition percentage of the compound on hERG potassium current, and I and Io represent the amplitude of hERG potassium current before and after drug addition, respectively.

[0400] Compound IC 50 Calculated using GraphPad Prism 5 software by fitting the following equation:

[0401] Y=Bottom+(Top-Bottom) / (1+10^((LogIC50-X)*HillSlope))

[0402] Where X is the Log value of the test sample concentration, Y is the inhibition percentage at the corresponding concentration, and Bottom and Top are the minimum and maximum inhibition percentages, respectively.

[0403] Table 6 IC of the compounds' inhibitory effects on hERG potassium channel current50

[0404] Conclusion: The compounds of the present invention, such as the compounds in the examples, have no significant inhibitory effect on hERG potassium channel current.

[0405] 9. CYP450 enzyme inhibition test

[0406] The purpose of this study was to evaluate the effects of test substances on the activities of five isoenzymes of cytochrome P450 (CYP) in human liver microsomes (CYP1A2, CYP2C9, CYP2C19, CYP2D6, and CYP3A4) using an in vitro test system. Specific probe substrates for the CYP450 isoenzymes were incubated with human liver microsomes and varying concentrations of the test substances. Reduced nicotinamide adenine dinucleotide phosphate (NADPH) was added to initiate the reaction. After the reaction, the samples were processed and the metabolites produced by the specific substrates were quantitatively detected by liquid chromatography-tandem mass spectrometry (LC-MS / MS). Changes in CYP enzyme activity were measured, and the IC values ​​were calculated. 50 The inhibitory potential of the test substance on each CYP enzyme isoform was evaluated.

[0407] Conclusion: The compounds of the present invention, such as the compounds in the examples, have no significant inhibitory activity on the five isoenzymes of human liver microsomal cytochrome P450 (CYP).

[0408] 10. Liver microsome stability test

[0409] In this study, liver microsomes from five species, including humans, dogs, rats, and mice, were used as in vitro models to evaluate the metabolic stability of the test substances.

[0410] At 37°C, 1 μM of the test substance was incubated with microsomal proteins and coenzyme NADPH. After a certain time (5, 10, 20, 30, 60 min), ice-cold acetonitrile containing the internal standard was added to terminate the reaction. The concentration of the test substance in the sample was detected by LC-MS / MS. The T1 / 2 was calculated based on the ln value of the drug residual rate in the incubation system and the incubation time, and the liver microsomal intrinsic clearance CLint(mic) and liver intrinsic clearance CLint(Liver) were further calculated.

[0411] Conclusion: The compounds of the present invention, such as the compounds in the examples, showed good metabolic stability in the liver microsome stability test.

[0412] 11. Caco-2 permeability test

[0413] The experiment used Caco-2 cell monolayers in 96-well Transwell plates, incubated in triplicate. Transport buffer (HBSS, 10 mM HEPES, pH 7.4 ± 0.05) containing the compound of the invention (2 μM) or the control compounds digoxin (10 μM), nadolol (2 μM), and metoprolol (2 μM) was added to the apical or basolateral wells of the cell monolayer. Transport buffer containing DMSO was added to the corresponding receiving wells. After incubation at 37 ± 1°C for 2 hours, the cell plate was removed and appropriate samples were taken from the apical and basolateral wells to a new 96-well plate. Proteins were then precipitated by adding acetonitrile containing an internal standard. Samples were analyzed using LC-MS / MS to determine the concentrations of the compound of the invention and the control compound. The concentration data were used to calculate the apparent permeability coefficients for transport from the apical to basolateral side of the cell monolayer and from the basolateral to apical side, thereby calculating the efflux rate. The integrity of the cell monolayer after 2 hours of incubation was assessed by leakage of Lucifer Yellow.

[0414] Conclusion: The compounds of the present invention, such as the compounds in the examples, showed good permeability in the Caco-2 permeability test.

[0415] 12. PK / PD testing in Dmd (Mdx) gene mutation mouse models

[0416] Objective: B10-Dmd-KO (Mdx) mice harbor a frameshift mutation in the Dmd gene, resulting in a loss-of-function Duchenne muscular dystrophy (DMD). Phenotypic analysis has shown that B10-Dmd-KO mice share a range of characteristics seen in DMD patients. This study investigated the effects of compound administration on plasma CK and TNNI2 levels in Mdx mice.

[0417] Experimental animals: B10-Dmd-KO (C57BL / 10ScSnJGpt-Dmdem3Cd4 / Gpt), male, 5-7 weeks old, weighing 28-30 g, supplier: Jiangsu Jicui Yaokang Biotechnology Co., Ltd.

[0418] Experimental process:

[0419] 1) Running training (D-5): Mdx mice were trained in batches with three training sessions: an initial speed of 4 m / min for 2 min, followed by 8 m / min for 8 min, and finally 12 m / min for 30 min, for a total of 40 min.

[0420] 2) Grip strength test (D-2): three times per round, three rounds of testing;

[0421] 3) Grouping (D0): After animals were anesthetized with isoflurane (concentration: 5%, gas flow rate: 1 L / min), blood (approximately 150 μL) was collected from the orbital venous sinus using a capillary tube into a 0.6 mL centrifuge tube containing EDTA anticoagulant. Plasma was separated and the CK value was measured, which was used as the baseline value for grouping.

[0422] 4) Administration, Grip, and Blood Collection (D1): The compound was dissolved in a solvent and orally administered to mice. The grip strength test was then initiated (three rounds of testing, three times each). One hour after the test, the animals were anesthetized with isoflurane (concentration: 5%, gas flow rate: 1 L / min) and blood (approximately 300 μL) was collected via the orbital venous sinus using a capillary tube into a 0.6 mL centrifuge tube containing EDTA anticoagulant (blood collection was performed 6 hours after administration).

[0423] 5) Running, Blood Collection, and Sample Collection (D2): 22 hours and 20 minutes after administration, run for 40 minutes (running parameters are the same as above). One hour after the completion of the run, the mice were anesthetized by inhalation of isoflurane (concentration: 5%, gas flow rate: 1 L / min). Blood (≥300 μL) was collected through the orbital venous sinus using a capillary tube into a 0.6 mL centrifuge tube containing EDTA anticoagulant. The animals were then euthanized, and the tibialis anterior muscle (left leg), soleus muscle (both legs), and myocardium were weighed and recorded. The blood was placed in a homogenate tube and quickly frozen in liquid nitrogen and stored at -80°C.

[0424] Sample Preparation: EDTA-anticoagulated whole blood was centrifuged at 3000 rpm and 4°C for 10 minutes, and serum was collected and aliquoted. The separated serum was analyzed using a biochemical analyzer (Roche C311 fully automated biochemical analyzer) for CK and TNNI2 levels (Mouse TNNI2 Elisa Kit, ABBEXA, E2207370Y). Serum concentrations of the compound, as well as drug concentrations in the tibialis anterior, soleus, and myocardial muscles, were determined using LC / MS.

[0425] Conclusion: The compounds of the present invention, such as the compounds in the examples, have a good inhibitory effect on the CK value in plasma in the Dmd (Mdx) gene mutation mouse model.

Claims

1. A compound represented by formula (I) or formula (I-1), a stereoisomer or a pharmaceutically acceptable salt thereof, in, R is selected from -O-halogenated C 1-4 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, -O-(CH2) r -R a 、C 3-10 Cycloalkyl, 5-10 membered heteroaryl containing 1-3 heteroatoms selected from N, O, S, -C 1-4 Alkyl-R a 、-C 1-4 Alkyl-OR a 、-C 1-4 Alkyl-NR b R a 、-NR b -S(O)2-R a 、-NR b -S(O)2-NR b R a 、-O-NR b R a 、-NH-OR b 、-NH-halogenated C 1-4 Alkyl, -OC 1-4 Alkyl, C 1-6 Alkoxy, halogen, cyano, nitro, C 1-4 Alkyl, -NH-C 1-4 Alkyl, -N(C 1-4 Alkyl)2, -NH-C 3-10 Cycloalkyl, -NHC(O)C 1-4 Alkyl, 4-10 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, S, -CO-C 1-4 Alkyl, -CO-R a 、-S(O)-R a 、-S(O)2-R a The alkyl, alkoxy, alkenyl, alkynyl, heteroaryl, cycloalkyl, heterocycloalkyl may be further substituted by 1-3 halogen, D, CN, OH, C 1-4 Alkyl, halogenated C 1-4 Alkyl, deuterated C 1-4 Alkyl, C 1-4 Alkoxy, halogenated C 1-4 Alkoxy, deuterated C 1-4 Alkoxy, C 3-10 substituted by cycloalkyl, 4-6 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, S, phenyl and NH2; R a Selected from CN, C 3-10 Cycloalkyl, 4-10 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, S, 6-10 membered aryl, 5-10 membered heteroaryl containing 1-3 heteroatoms selected from N, O, S, -C(O)-R a1 The cycloalkyl, heterocycloalkyl, aryl, heteroaryl groups are optionally further substituted by 1-3 groups selected from halogen, =O, deuterium, CN, OH, C 1-4 Alkyl, halogenated C 1-4 Alkyl, deuterated C 1-4 Alkyl, C 1-4 Alkoxy, halogenated C 1-4 Alkoxy, deuterated C 1-4 Alkoxy and NH2 group substitution; Each R b Selected from H, deuterium, C 1-4 Alkyl, C 3-6 Cycloalkyl, halogenated C 1-4 Alkyl, deuterated C 1-4 alkyl; R a1 Selected from OH, NH2, -NHC 1-4 Alkyl, -N(C 1-4 Alkyl)2, -NHC 3-10 Cycloalkyl, C 1-4 Alkoxy, C 1-6 Alkyl, C 3-10 Cycloalkyl, 6-10 membered aryl, 5-10 membered heteroaryl containing 1-3 heteroatoms selected from N, O, S; R 1 、R 2 、R 3 、R 4 、R 5 、R 6 、R 8 、R 9 Each independently selected from H, deuterium, halogen, OH, C 1-4 Alkyl, halogenated C 1-4 Alkyl, deuterated C 1-4 Alkyl, C 1-4 Alkoxy, halogenated C 1-4 Alkoxy, deuterated C 1-4 Alkoxy, C 3-6 Cycloalkyl, or R 1 and R 3 , or R 4 and R 5 , or R 3 and R 4 Together with the carbon atom to which it is attached, it forms C 3-6 Cycloalkyl, optionally further substituted by 1-3 groups selected from =O, Halogen, deuterium, CN, OH, C 1-3 Alkyl, halogenated C 1-3 Alkyl, deuterated C 1-3 Alkyl, C 1-3 Alkoxy, halogenated C 1-3 Alkoxy, deuterated C 1-3 Alkoxy group substitution; Alternatively, R 1 and R 2 、R 3 and R 4 、R 5 and R 6 Together with the carbon atom to which it is attached, it forms =O, =CH2, =CF2 or a 5-10 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, S, which is optionally further substituted by 1-3 heteroatoms selected from =O, halogen, deuterium, CN, OH, C 1-3 Alkyl, halogenated C 1-3 Alkyl, deuterated C 1-3 Alkyl, C 1-3 Alkoxy, halogenated C 1-3 Alkoxy, deuterated C 1-3 Alkoxy group substitution; X is selected from N or CH; R 7 Selected from C 3-10 Cycloalkyl, 4-10 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, S, 6-10 membered aryl, 5-10 membered heteroaryl containing 1-3 heteroatoms selected from N, O, S, wherein the cycloalkyl, heterocycloalkyl, aryl, heteroaryl may be further substituted by 1-3 halogens, =O, deuterium, CN, OH, C 1-4 Alkyl, halogenated C 1-4 Alkyl, deuterated C 1-4 Alkyl, C 1-4 Alkoxy, halogenated C 1-4 Alkoxy, deuterated C 1-4 Alkoxy, NH2, =CH2 and =CF2 group substitution; r is selected from 0, 1 or 2; And the compound of formula (I) is not the following structure:

2. The compound according to claim 1, its stereoisomer or pharmaceutically acceptable salt, R 1 、R 2 、R 3 、R 4 、R 6 、R 8 、R 9 Each independently selected from H, deuterium, halogen, OH, C 1-4 Alkyl, halogenated C 1-4 Alkyl, deuterated C 1- 4 alkyl, C 1-4 Alkoxy, halogenated C 1-4 Alkoxy, deuterated C 1-4 Alkoxy, C 3-6 Cycloalkyl, and R 1 and R 3 , or R 4 and R 5 , or R 3 and R 4 Together with the carbon atom to which it is connected, it forms a C3, C4, C5, or C6 cycloalkyl group, wherein the C3, C4, C5, or C6 cycloalkyl group is optionally further substituted by 1, 2, or 3 groups selected from ═O, Halogen, deuterium, CN, OH, C 1-3 Alkyl, halogenated C 1-3 Alkyl, deuterated C 1-3 Alkyl, C 1-3 Alkoxy, halogenated C 1-3 Alkoxy, deuterated C 1-3 Alkoxy groups are substituted.

3. The compound according to claim 1, its stereoisomer or pharmaceutically acceptable salt, R 1 、R 2 、R 3 、R 4 、R 8 、R 9 Each independently selected from H, deuterium, halogen, OH, C 1-4 Alkyl, halogenated C 1-4 Alkyl, deuterated C 1-4 Alkyl, C 1-4 Alkoxy, halogenated C 1-4 Alkoxy, deuterated C 1-4 Alkoxy, C 3-6 Cycloalkyl, or R 3 and R 4 Together with the carbon atom to which it is connected, it forms a C3, C4, C5, or C6 cycloalkyl group, wherein the C3, C4, C5, or C6 cycloalkyl group is optionally further substituted by 1, 2, or 3 groups selected from ═O, Halogen, deuterium, CN, OH, C 1-3 Alkyl, halogenated C 1-3 Alkyl, deuterated C 1-3 Alkyl, C 1-3 Alkoxy, halogenated C 1-3 Alkoxy, deuterated C 1- 3 alkoxy group substitution; And R 5 、R 6 Together with the carbon atom to which it is attached, it forms =O, =CH2, =CF2 or a 5-10 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, S, which is optionally further substituted by 1-3 heteroatoms selected from =O, halogen, deuterium, CN, OH, C 1-3 Alkyl, halogenated C 1-3 Alkyl, deuterated C 1-3 Alkyl, C 1-3 Alkoxy, halogenated C 1-3 Alkoxy, deuterated C 1-3 Alkoxy groups are substituted.

4. The compound according to claim 1, wherein the stereoisomer or pharmaceutically acceptable salt thereof has the structure of formula (II) or (II-1): R is selected from -O-halogenated C 1-4 Alkyl, -O- containing 1-3 heteroatoms selected from N, O, S 4-10 membered heterocycloalkyl, the alkyl, heterocycloalkyl optionally further 1-3 selected from halogen, D, CN, OH, C 1-4 Alkyl, halogenated C 1-4 Alkyl, deuterated C 1-4 Alkyl, C 1-4 Alkoxy, halogenated C 1-4 Alkoxy, deuterated C 1-4 Alkoxy and NH2 groups are substituted.

5. The compound according to claim 1, its stereoisomer or pharmaceutically acceptable salt, wherein R is selected from -COCH3、 -COCF3、-COCH3CF3、 R 1 、R 2 、R 3 、R 4 、R 5 、R 6 、R 8 、R 9 Each independently selected from H, deuterium, C 1-2 Alkyl, halogenated C 1-2 Alkyl, C 3-6 Cycloalkyl; or R 1 and R 3 , or R 3 and R 4 Together with the carbon atom to which it is attached, it forms C 3-6 Cycloalkyl, said cycloalkyl optionally further substituted by 1-3 groups selected from =O, Halogen, deuterium, C 1-3 Alkyl, halogenated C 1-3 Alkyl, deuterated C 1-3 Alkyl, C 1-3 Alkoxy, halogenated C 1-3 Alkoxy, deuterated C 1-3 Alkoxy group substitution; Alternatively, R 5 and R 6 Together with the carbon atom to which it is attached, it forms =O, =CH2, =CF2 or a 5-10 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, S, which is optionally further substituted by 1-3 heteroatoms selected from =O, halogen, deuterium, CN, OH, C 1-3 Alkyl, halogenated C 1-3 Alkyl, deuterated C 1-3 Alkyl, C 1-3 Alkoxy, halogenated C 1-3 Alkoxy, deuterated C 1-3 Alkoxy group substitution; Alternatively, R 1 and R 2 、R 3 and R 4 Together with the carbon atom to which it is attached, it forms =O, =CH2, or =CF2; The condition is that when R 5 and R 6 Together with the carbon atom to which it is attached, it forms =O, R 1 、R 2 、R 3 、R 4 、R 8 、R 9 When selected from H, R is not selected from the following structures:

6. The compound according to claim 5, its stereoisomer or pharmaceutically acceptable salt, wherein R 1 、R 2 、R 5 、R 6 、R 8 、R 9 selected from H or deuterium; R 3 、R 4 Selected from H, deuterium, C 1-2 alkyl; or R 1 and R 3 , or R 3 and R 4 Together with the carbon atom to which it is attached, it forms C 3-6 Cycloalkyl, the cycloalkyl is optionally further substituted by 1-3 groups selected from Halogen, deuterium, C 1-3 Alkyl, halogenated C 1-3 Alkyl, deuterated C 1-3 Alkyl radical substitution; Alternatively, R 5 and R 6 Together with the carbon atom to which it is attached, it forms =O, =CH2, =CF2 or a 5-6 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, S, which is optionally further substituted by 1-3 heteroatoms selected from halogen, deuterium, C 1-3 Alkyl, halogenated C 1-3 Alkyl, deuterated C 1-3 Alkyl groups are substituted.

7. The compound according to claim 1, its stereoisomers or pharmaceutically acceptable salts, wherein the compound is selected from one of the structures in Table 1 or Table 2.

8. A pharmaceutical composition or pharmaceutical preparation comprising the compound according to any one of claims 1 to 7, or a stereoisomer or pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier and / or excipient.

9. The pharmaceutical composition or pharmaceutical preparation according to claim 8, comprising 1-1500 mg of the compound or stereoisomer or pharmaceutically acceptable salt according to any one of claims 1-7 and a carrier and / or excipient.

10. Use of the compound according to any one of claims 1 to 7, its stereoisomers or pharmaceutically acceptable salts, or the pharmaceutical composition or pharmaceutical preparation according to any one of claims 8 to 9 in the preparation of a medicament for treating / preventing Myosin II-mediated diseases. The use according to claim 10 , wherein the Myosin II-mediated disease is selected from muscular dystrophy.

12. A method for treating a disease in a mammal, comprising administering to a subject a therapeutically effective amount of the compound or stereoisomer or pharmaceutically acceptable salt according to any one of claims 1 to 7, preferably 1 to 1500 mg, wherein the disease is preferably muscular dystrophy.

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