Myosin II inhibitor and application thereof

Abstract

The invention discloses a compound shown as a formula (I), or a stereoisomer, a pharmaceutically acceptable salt and a pharmaceutical composition of the compound and the stereoisomer and the pharmaceutically acceptable salt, and application of the compound and the stereoisomer and the pharmaceutically acceptable salt in preparation of drugs for treating / preventing Myosin II mediated diseases, and all groups in the formula (I) are defined as in the specification.

Description

TECHNICAL FIELD

[0001] The present application relates to a Myosin II inhibitor, stereoisomers, pharmaceutically acceptable salts thereof and use thereof in the manufacture of a medicament for treating Myosin II mediated related diseases. BACKGROUND

[0002] Skeletal muscle has two major roles that are critical to the human body: 1) muscle contraction, which generates movement and maintains posture; and 2) skeletal muscle is also the site of glucose, fatty acid, and amino acid metabolism. In a normal human body during daily activities, skeletal muscle contraction is closely associated with muscle stress, breakdown, and remodeling, which are critical for muscle adaptation. However, in patients with progressive muscular dystrophy, such as Duchenne muscular dystrophy (DMD), muscle contraction leads to muscle breakdown that is difficult to repair, amplified in a continuous cycle. As the patient ages, these changes gradually accumulate and develop into a pathological process, which leads to excessive inflammation, fibrosis, and accumulation of fat deposits in the muscle, which in turn progresses to a sharp decline in physical function, ultimately leading to death.

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

[0004] The present application provides a compound of formula (I-1), (I-2), (I-3), (I-4), (I-5), (I-6), (I-7), (I-8), (I-9), (I-10), (I-11), (I-12), (I-13), (I-14), (I-15), (I-16), (II), (III), stereoisomers, pharmaceutically acceptable salts thereof, which has the excellent effects of good activity, excellent physicochemical properties, easy preparation, excellent pharmacokinetic properties, high bioavailability, and low toxicity and side effects.

[0005] The compound of formula (I-1), (I-2), (I-3), (I-4), (I-5), (I-6), (I-7), (I-8), (I-9), (I-10), (I-11), (I-12), (I-13), (I-14), (I-15), (I-16), (II), (III), stereoisomers, pharmaceutically acceptable salts thereof,

[0006]

[0007]

[0008] wherein,

[0009] represents a single or double bond;

[0010] m is selected from 0, 1, 2; in some embodiments, m is selected from 0, 1; in some embodiments, m is selected from 1; in some embodiments, m is selected from 0;

[0011] p is selected from 0, 1, 2; in some embodiments, p is selected from 0, 1; in some embodiments, p is selected from 1; in some embodiments, p is selected from 0;

[0012] n is selected from 0, 1, 2; in some embodiments, n is selected from 0, 1, 2; in some embodiments, n is selected from 0, 1; in some embodiments, n is selected from 1; in some embodiments, n is selected from 0;

[0013] A is selected from 4-10 membered heterocycloalkyl, 5-10 membered heteroaryl, 6-10 membered aryl;

[0014] In some embodiments, A is selected from 6-10 membered heterocycloalkyl, 5-6 membered heteroaryl, 9-10 membered heteroaryl, phenyl, naphthyl;

[0015] In some embodiments, A is selected from

[0016] In some embodiments, A is selected from

[0017] each R 1 , R 2 is each independently selected from H, =O, C 1-4 alkyl, haloC 1-4 alkyl, -O-haloC 1-4 alkyl, -NH-haloC 1-4 alkyl, -OC 1-4 alkyl, C 1-4 alkoxy, halogen, cyano, nitro, OH, C 1-4 alkyl, C 2-6 alkenyl, -NH-C 1-4 alkyl, -N(C 1-4 alkyl)2, -NH-C 3-10 cycloalkyl, -NHC(O)C 1-4 alkyl, C 3-10 cycloalkyl, -O-C 3-10 cycloalkyl, -O-(4-10 membered heterocycloalkyl), -O-C 1-4 alkyl-C3-10 Cycloalkyl, amino, or 4-10 membered heterocyclic alkyl, wherein the alkyl, alkoxy, cycloalkyl, or heterocyclic alkyl may optionally be further selected from 1-3 halogens, 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 Substitution with cycloalkyl, 4-6 membered heterocyclic alkyl, phenyl, and NH2 groups;

[0018] In some implementation schemes, R 1 Selected from H, =O, C 1-4 Alkyl, C 1-4 Alkoxy, halogenated C 1-4 Alkyl, -O-halogenated C 1-4 Alkyl, C 3-6 cycloalkyl, -OC 3-6 Cycloalkyl, 4-7 membered heterocycloalkyl, -O-(4-7 membered heterocycloalkyl), -OC 1-4 Alkyl-C 3-6 cycloalkyl, -OC 1-4 Alkyl group (4-7-membered heterocyclic alkyl group), wherein the alkyl, cycloalkyl, or heterocyclic alkyl group is optionally further selected from 1-3 halogens, 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 3-5 Substitution with cycloalkyl, 4-6 membered heterocyclic alkyl, phenyl, and NH2 groups;

[0019] In some implementation schemes, R 1 Selected from H, =O, C 1-2 Alkyl, C 1-2 Alkoxy, halogenated C 1-2 Alkyl, -O-halogenated C 1-2 Alkyl, cyclopropyl, cyclobutyl, -O-cyclopropyl, -O-cyclobutyl, -O-methylene-cyclopropyl, -O-methylene-cyclobutyl, -O-oxecyclopentyl, wherein the alkyl, alkoxy, cyclopropyl, cyclobutyl, and oxecyclopentyl groups are optionally further selected from 1-3 halogens, D, CN, OH, and C. 1-2 Alkyl group substitution;

[0020] In some implementation schemes, R 1 Selected from

[0021] In some implementations, each R 2 Each is independently selected from =O, C 2-4 alkenyl, C 2-4 Alkyne, halogen, cyano, nitro, amino, -NH-C 1-4 Alkyl, -N(C) 1-4 Alkyl)2, OH, C 1-4 Alkyl, Halogenated C 1-4 Alkyl, deuterated C 1-4 Alkyl, C 3-6 Cycloalkyl, 4-7 membered heterocyclic alkyl;

[0022] In some implementations, each R 2 Each is independently selected from =O, C 2-4 alkenyl, C 2-4 Alkyne, halogen, cyano, nitro, amino, -NH-C 1-4 Alkyl, -N(C) 1-4 Alkyl)2, OH, C 1-4 Alkyl, Halogenated C 1-4 Alkyl, deuterated C 1-4 Alkyl, C 3-6 Cycloalkyl, 4-7 membered heterocyclic alkyl;

[0023] In some implementations, each R 2 Each group is independently selected from =O, vinyl, propenyl, ethynyl, propynyl, halogen, cyano, nitro, amino, -NH-C 1-2 Alkyl, -N(C) 1-2 Alkyl)2, OH, C 1-2 Alkyl, Halogenated C 1-2 Alkyl, deuterated C 1-2 Alkyl, cyclopropyl, cyclobutyl, azircyclopentyl, oxacyclopentyl;

[0024] Or, R 1 and R 2 or two adjacent R 2 Together with the linked atoms, they form a 5-6 membered carbon ring or a 5-6 membered heterocycle, wherein the carbon ring or heterocycle may optionally be further composed of 1-3 atoms selected from halogens, D, CN, OH, and 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, -OC 3-10 Substitution with cycloalkyl, 4-6 membered heterocyclic alkyl, phenyl, and NH2 groups;

[0025] In some implementation schemes, Selected from

[0026]

[0027] in, This indicates the site linked to L1, where L1 is a bond. Indicates the site connected to ring B;

[0028] In some implementation schemes, Selected from

[0029] B is selected from 5-6-membered heteroaryl, 8-10-membered heteroaryl, and 5-6-membered heterocyclic alkyl;

[0030] In some embodiments, ring B is selected from pyridazinones, pyridinones, pyrazinones, pyrimidinones, etc. 5-membered heteroaryl, 8-10-membered bicyclic heteroaryl;

[0031] In some implementation schemes, ring B is selected from...

[0032] Each R 4 Each is independently selected from C 3-10 cycloalkyl, C 2-6 alkenyl, C 2-6 Alkyne, halogen, =O, amino, cyano, -NH-C 1-4 Alkyl, -N(C) 1-4 Alkyl)2, C 1-4 Alkyl, C 1-4 Alkoxy, halogenated C 1-4 Alkyl, 4-10 membered heterocyclic alkyl, 5-10 membered heteroaryl, 6-10 membered aryl;

[0033] In some implementations, each R 4 Each is independently selected from deuterium, =O, halogen, cyano, and 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;

[0034] In some implementation schemes, R 4 Selected from deuterium, =O, halogen, cyano, C 1-2 Alkyl, Halogenated C 1-2 Alkyl, deuterated C 1-2 Alkyl, C 1-2 Alkoxy, halogenated C1-2 Alkoxy, deuterated C 1-2 Alkoxy;

[0035] In some implementation schemes, Selected from

[0036] * indicates the linking site between loop B and L2. When L2 is a bond, * indicates the linking site between loop B and C. ** indicates the linking site between loop B and L1. When L1 is a bond, ** indicates the linking site between loop B and loop A.

[0037] C is selected from C 3-8 Alkyl, phenyl, 5-6 membered heteroaryl, C 3-12 Carbocyclic rings, 4-15 membered heterocyclic groups;

[0038] In some implementations, C is selected from C 5-7 Alkyl, phenyl, 5-6 membered heteroaryl, C 3-10 Carbocyclic rings, 4-14 membered heterocyclic groups;

[0039] In some implementations, C is selected from C 5-7 alkyl,

[0040] Each X is independently selected from CR 6 Or N;

[0041] In some implementations, X is selected from CH or N;

[0042] R 7 Selected from H, halogen, SF5, OH, CN, amino, C 1-4 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 1-4 Alkoxy, halogenated C 1-4 Alkyl, =O, =CH2, =CF2, =C(CH3)2, -NH-C 1-4 Alkyl, -N(C) 1-4 Alkyl)2、-S(O)R c -S(O)2R c -C(O)R c -C(O)OR c -C(O)N(R) c )2、-OC(O)R c -C 0-4 Alkyl-(4-10 membered heterocyclic alkyl), or -C 0-4 Alkyl-(5-10-membered heteroaryl), -P(O)(R c )2、-Si(R c)3、-SF5、-S(O)(=NH)R c The alkyl, alkenyl, alkynyl, alkoxy, heterocyclic alkyl, and heteroaryl groups may optionally be further selected from 1-3 halogens, deuterium, CN, OH, and 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 Substitution of alkoxy and NH2 groups;

[0043] In some implementation schemes, R 7 Selected from H, halogens, 5-6 quinone heteroaryl groups, C 1-4 Alkyl, C 1-4 Alkoxy, C 2-4 alkenyl, C 2-4 alkynyl, halogenated C 1-4 Alkyl, -C(O)C 1-4 Alkyl, =O, =CH2, =CF2, =C(CH3)2, wherein the alkyl, alkenyl, alkynyl, alkoxy, or heteroaryl groups are optionally further selected from 1-3 halogens, 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 Substitution of alkoxy and NH2 groups;

[0044] In some implementation schemes, R 7 Selected from H, halogens, 5-membered heteroaryl groups, C 1-2 Alkyl, C 1-2 Alkoxy, C 2-4 alkenyl, C 2-4 alkynyl, halogenated C 1-2 Alkyl, -C(O)C 1-2 Alkyl, =O, =CH2, =CF2, =C(CH3)2, wherein the alkyl, alkenyl, alkynyl, alkoxy, or heteroaryl groups are optionally further selected from 1-3 halogens, 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 group substitution;

[0045] In some implementation schemes, R 7Selected from H, halogen, OH, NH2, CN, =O, =CH2, =CF2, =C(CH3)2, methyl, ethyl, trifluoromethyl, cyclopropyl;

[0046] Each R 6 Each element is independently selected from H, halogen, SF5, OH, CN, amino, and C. 1-4 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 1-4 Alkoxy, halogenated C 1-4 Alkyl, -NH-C 1-4 Alkyl, -N(C) 1-4 Alkyl)2、-S(O)R c -S(O)2R c -C(O)R c -C(O)OR c -C(O)N(R) c )2、-OC(O)R c The alkyl, alkenyl, alkynyl, and alkoxy groups may optionally be further selected from 1-3 groups selected from halogen, deuterium, CN, OH, and 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 Substitution of alkoxy and NH2 groups;

[0047] In some implementations, each R 6 Each element is independently selected from deuterium, halogens, OH, CN, and 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 Alkyl groups, SF5;

[0048] In some implementations, each R 6 Each element is independently selected from deuterium, halogens, OH, CN, and 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 Alkyl groups, SF5;

[0049] In some implementation schemes, R 6 Selected from halogens;

[0050] Each Rc Each is independently selected from H, OH, and C. 1-4 Alkyl, C 3-7 Cyclopropyl, Halogenated C 1-4 Alkyl, deuterated C 1-4 Alkyl, C 1-4 Alkoxy, halogenated C 1-4 Alkoxy, deuterated C 1-4 Alkoxy groups, NH2;

[0051] L1 is selected from key, C 2-6 alkenyl, C 2-6 Alkyne, O, NH, S, -CD2-, -CHD-, -CR L1 R L2 -、C 1-4 Alkyl, -C(O)-, S(O), S(O)2, wherein the alkyl group is optionally further modified by R L1 replace;

[0052] In some embodiments, L1 is selected from the group of bonds and ethynyl groups; in other embodiments, L1 is selected from the group of bonds.

[0053] L2 is selected from bonds, O, NH, S, -CH2-, -CD2-, -CHD-, -CR L1 R L2 -、-CHR L2 -、-CDR L2 -, -CH2C(R) L1 =, -CH2CH=CH-CH=, C 2-4 Alkyl, C 2-4 alkenyl, C 2-4 Alkyne group, -C(O)-, S(O), S(O)2, wherein the alkyl group is optionally further surrounded by 1-3 R groups. L1 replace;

[0054] In some implementations, L2 is selected from the bond, -CH2-, -CH2CH2-, -C(O)-, -CHF-, -CF2-, C(CH3)2-, ethynyl, propynyl; * indicates the site where L2 is connected to C;

[0055] In some implementations, L2 is selected from the bond, -CH2-, -CH2CH2-, -C(O)-, -CHF-, -CF2-, C(CH3)2-, propynyl; * indicates the site where L2 is connected to C;

[0056] R L1 R L2 Each element is independently selected from H, halogen, OH, CN, amino, and C. 1-4 Alkyl, C 1-4Alkoxy, C 3-6 Cycloalkyl groups, and 4-6 membered heterocyclic alkyl groups containing 1-3 heteroatoms selected from N, O, and S;

[0057] In some implementation schemes, R L1 R L2 Each element is independently selected from H, halogen, OH, CN, amino, and C. 1-2 Alkyl, C 1-2 Alkoxy, C 3-6 Cycloalkyl, 4-6 membered heterocyclic alkyl;

[0058] In some implementation schemes, R L1 R L2 Each is independently selected from H, F, Cl, OH, CN, amino, methyl, methoxy, cyclopropyl, and cyclobutyl;

[0059] In some implementation schemes, Selected from

[0060] In some implementation schemes, Selected from

[0061] In some implementation schemes, Selected from

[0062]

[0063] or Selected from

[0064] As an option, R 2 R 4 L1 and the attached atoms together form a 5-7 membered carbon ring and a 5-7 membered heterocycle, wherein the carbon ring and heterocycle are optionally further composed of 1-3 atoms selected from R. A3 Substitution of groups;

[0065] As an option, R 4 R 6 L2 and the attached atoms together form a 5-7 membered carbon ring and a 5-7 membered heterocycle, wherein the carbon ring and heterocycle are optionally further composed of 1-3 atoms selected from R A3 Substitution of groups;

[0066] As an option, two R 6 Substituents, together with the atoms they are attached to, form 4-6 membered carbon rings, 4-6 membered heterocycles, or R 6 R 7Together with the attached atoms, they form 4-12 membered carbon rings and 4-12 membered heterocycles, wherein the carbon rings and heterocycles are optionally further composed of 1-3 atoms selected from R A3 Substitution of groups;

[0067] Each R A3 Each element is independently selected from O, halogen, deuterium, CN, OH, and 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 groups, NH2;

[0068] In some implementations, each R A3 Each element is independently selected from O, halogen, deuterium, CN, OH, and 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 groups, NH2;

[0069] In some implementations, each R A3 Each is independently selected from =O, F, Cl, deuterium, CN, OH, methyl, trifluoromethyl, methoxy, halomethoxy, and NH2.

[0070] Specifically, the first technical solution of the present invention relates to a compound of formula (I), its stereoisomers, and pharmaceutically acceptable salts.

[0071]

[0072] in,

[0073] Indicates a single bond or a double bond;

[0074] m is selected from 0, 1, and 2;

[0075] p is selected from 0, 1, and 2;

[0076] n is selected from 0, 1, and 2;

[0077] A is selected from 4-10-membered heterocyclic alkyl, 5-10-membered heteroaryl, and 6-10-membered aryl;

[0078] B is selected from 5-6-membered heteroaryl, 8-10-membered heteroaryl, and 5-6-membered heterocyclic alkyl;

[0079] C is selected from C 3-8 Alkyl, phenyl, 5-6 membered heteroaryl, C 3-12Carbocyclic rings, 4-15 membered heterocyclic groups;

[0080] Each R 1 R 2 Each is independently selected from H, =O, C 1-4 Alkyl, Halogenated C 1-4 Alkyl, -O-halogenated C 1-4 Alkyl, -NH-halogenated C 1-4 Alkyl, -OC 1-4 Alkyl, C 1-4 Alkyl group, halogen, cyano group, nitro group, OH, C 1-4 Alkyl, C 2-6 alkenyl, -NH-C 1-4 Alkyl, -N(C) 1-4 Alkyl)2、-NH-C 3-10 cycloalkyl, -NHC(O)C 1-4 Alkyl, C 3-10 cycloalkyl, -OC 3-10 Cycloalkyl, -O- (4-10 membered heterocycloalkyl), -OC 1-4 Alkyl-C 3-10 Cycloalkyl, amino, or 4-10 membered heterocyclic alkyl, wherein the alkyl, alkoxy, cycloalkyl, or heterocyclic alkyl may optionally be further selected from 1-3 halogens, 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 Substitution with cycloalkyl, 4-6 membered heterocyclic alkyl, phenyl, and NH2 groups;

[0081] Or, R 1 and R 2 or two adjacent R 2 Together with the linked atoms, they form a 5-6 membered carbon ring or a 5-6 membered heterocycle, wherein the carbon ring or heterocycle may optionally be further composed of 1-3 atoms selected from halogens, D, CN, OH, and 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, -OC 3-10 Substitution with cycloalkyl, 4-6 membered heterocyclic alkyl, phenyl, and NH2 groups;

[0082] Each R 4 Each is selected independently from C 3-10cycloalkyl, C 2-6 alkenyl, C 2-6 Alkyne, halogen, =O, amino, cyano, -NH-C 1-4 Alkyl, -N(C) 1-4 Alkyl)2, C 1-4 Alkyl, C 1-4 Alkoxy, halogenated C 1-4 Alkyl, 4-10 membered heterocyclic alkyl, 5-10 membered heteroaryl, 6-10 membered aryl;

[0083] R 7 Selected from H, halogen, SF5, OH, CN, amino, C 1-4 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 1-4 Alkoxy, halogenated C 1-4 Alkyl, =O, =CH2, =CF2, =C(CH3)2, -NH-C 1-4 Alkyl, -N(C) 1-4 Alkyl)2、-S(O)R c -S(O)2R c -C(O)R c -C(O)OR c -C(O)N(R) c )2、-OC(O)R c -C 0-4 Alkyl-(4-10 membered heterocyclic alkyl), or -C 0-4 Alkyl-(5-10-membered heteroaryl), -P(O)(R c )2、-Si(R c )3、-SF5、-S(O)(=NH)R c The alkyl, alkenyl, alkynyl, alkoxy, heterocyclic alkyl, and heteroaryl groups may optionally be further selected from 1-3 halogens, deuterium, CN, OH, and 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 Substitution of alkoxy and NH2 groups;

[0084] Each R 6 Each element is independently selected from H, halogen, SF5, OH, CN, amino, and C. 1-4 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 1-4 Alkoxy, halogenated C 1-4 Alkyl, -NH-C 1-4 Alkyl, -N(C)1-4 Alkyl)2、-S(O)R c -S(O)2R c -C(O)R c -C(O)OR c -C(O)N(R) c )2、-OC(O)R c The alkyl, alkenyl, alkynyl, and alkoxy groups may optionally be further selected from 1-3 groups selected from halogen, deuterium, CN, OH, and 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 Substitution of alkoxy and NH2 groups;

[0085] Each R c Each is independently selected from H, OH, and C. 1-4 Alkyl, C 3-7 Cyclopropyl, Halogenated C 1-4 Alkyl, deuterated C 1-4 Alkyl, C 1-4 Alkoxy, halogenated C 1-4 Alkoxy, deuterated C 1-4 Alkoxy groups, NH2;

[0086] L1 is selected from key, C 2-6 alkenyl, C 2-6 Alkyne, O, NH, S, -CD2-, -CHD-, -CR L1 R L2 -、C 1-4 Alkyl, -C(O)-, S(O), S(O)2, wherein the alkyl group is optionally further modified by R L1 replace;

[0087] L2 is selected from bonds, O, NH, S, -CH2-, -CD2-, -CHD-, -CR L1 R L2 -、-CDR L2 -, -CH2C(R) L1 =, -CH2CH=CH-CH=, C 2-4 Alkyl, C 2-4 alkenyl, C 2-4 Alkyne group, -C(O)-, S(O), S(O)2, wherein the alkyl group is optionally further surrounded by 1-3 R groups. L1 replace;

[0088] R L1 R L2 Each element is independently selected from halogen, OH, CN, amino, C1-4 Alkyl, C 1-4 Alkoxy, C 3-6 Cycloalkyl, 4-6 membered heterocyclic alkyl;

[0089] As an option, R 2 R 4 L1 and the attached atoms together form a 5-7 membered carbon ring and a 5-7 membered heterocycle, wherein the carbon ring and heterocycle are optionally further composed of 1-3 atoms selected from R. A3 Substitution of groups;

[0090] As an option, R 4 R 6 L2 and the attached atoms together form a 5-7 membered carbon ring and a 5-7 membered heterocycle, wherein the carbon ring and heterocycle are optionally further composed of 1-3 atoms selected from R A3 Substitution of groups;

[0091] As an option, two R 6 Substituents, together with the atoms they are attached to, form 4-6 membered carbon rings, 4-6 membered heterocycles, or R 6 R 7 Together with the attached atoms, they form 4-12 membered carbon rings and 4-12 membered heterocycles, wherein the carbon rings and heterocycles are optionally further composed of 1-3 atoms selected from R A3 Substitution of groups;

[0092] Each R A3 Each element is independently selected from O, halogen, deuterium, CN, OH, and 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 groups, NH2.

[0093] The second technical solution of the present invention is that the compound of formula (I) of the present invention, its stereoisomers and pharmaceutically acceptable salts, have the structures of formulas (I-1), (I-2), (I-3), (I-4), (I-5), (I-6), (I-7), (I-8), (I-9), (I-10), (I-11), (I-12), (I-13), (I-14), (I-15), and (I-16):

[0094]

[0095] Each X is independently selected from CR 6 Or N;

[0096] The definitions of other functional groups are consistent with any of the technical solutions mentioned above.

[0097] The third technical solution of the present invention is the compound of formula (I) described in the present invention, its stereoisomers, and pharmaceutically acceptable salts, wherein,

[0098] m is selected from 0 or 1;

[0099] p is selected from 0 and 1;

[0100] n is selected from 0 or 1;

[0101] A is selected from

[0102] R 1 Selected from H, =O, C 1-4 Alkyl, C 1-4 Alkoxy, halogenated C 1-4 Alkyl, -O-halogenated C 1-4 Alkyl, C 3-6 cycloalkyl, -OC 3-6 Cycloalkyl, 4-7 membered heterocycloalkyl, -O-(4-7 membered heterocycloalkyl), -OC 1-4 Alkyl-C 3-6 cycloalkyl, -OC 1-4 Alkyl group (4-7-membered heterocyclic alkyl group), wherein the alkyl, cycloalkyl, or heterocyclic alkyl group is optionally further selected from 1-3 halogens, 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 3-5 Substitution with cycloalkyl, 4-6 membered heterocyclic alkyl, phenyl, and NH2 groups;

[0103] Each R 2 Each is independently selected from =O, C 2-4 alkenyl, C 2-4 Alkyne, halogen, cyano, nitro, amino, -NH-C 1-4 Alkyl, -N(C) 1-4 Alkyl)2, OH, C 1-4 Alkyl, Halogenated C 1-4 Alkyl, deuterated C 1-4 Alkyl, C 3-6 Cycloalkyl, 4-7 membered heterocyclic alkyl;

[0104] Ring B is selected from pyridazinones, pyridinones, pyrazinones, and pyrimidinones. 5-membered heteroaryl, 8-10-membered bicyclic heteroaryl;

[0105] Each R 4 Selected from deuterium, =O, halogen, cyano, C1-4 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;

[0106] C is selected from C 5-7 alkyl,

[0107] Each R 6 Each element is independently selected from deuterium, halogens, OH, CN, and 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 Alkyl groups, SF5;

[0108] R 7 Selected from H, halogens, 5-6 quinone heteroaryl groups, C 1-4 Alkyl, C 1-4 Alkoxy, C 2-4 alkenyl, C 2-4 alkynyl, halogenated C 1-4 Alkyl, -C(O)C 1-4 Alkyl, =O, =CH2, =CF2, =C(CH3)2, wherein the alkyl, alkenyl, alkynyl, alkoxy, or heteroaryl groups are optionally further selected from 1-3 halogens, 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 Substitution of alkoxy and NH2 groups;

[0109] L1 is selected from the bond and ethynyl group;

[0110] L2 is selected from the bond, -CH2-, -CH2CH2-, -C(O)-, -CHF-, -CF2-, C(CH3)2-, ethynyl, propynyl; * indicates the site where L2 is connected to C.

[0111] The definitions of other functional groups are consistent with any of the technical solutions mentioned above.

[0112] The fourth technical solution of the present invention, the compound of formula (I) of the present invention, its stereoisomer, pharmaceutically acceptable salt, wherein,

[0113] R1 Selected from H, =O, C 1-2 Alkyl, C 1-2 Alkoxy, halogenated C 1-2 Alkyl, -O-halogenated C 1-2 Alkyl, cyclopropyl, cyclobutyl, -O-cyclopropyl, -O-cyclobutyl, -O-methylene-cyclopropyl, -O-methylene-cyclobutyl, -O-oxecyclopentyl, wherein the alkyl, alkoxy, cyclopropyl, cyclobutyl, and oxecyclopentyl groups are optionally further selected from 1-3 halogens, D, CN, OH, and C. 1-2 Alkyl group substitution;

[0114] Each R 2 Each group is independently selected from =O, vinyl, propenyl, ethynyl, propynyl, halogen, cyano, nitro, amino, -NH-C 1-2 Alkyl, -N(C) 1-2 Alkyl)2, OH, C 1-2 Alkyl, Halogenated C 1-2 Alkyl, deuterated C 1-2 Alkyl, cyclopropyl, cyclobutyl, azircyclopentyl, oxacyclopentyl;

[0115] R 4 Selected from deuterium, =O, halogen, cyano, 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;

[0116] Each R 6 Each element is independently selected from deuterium, halogens, OH, CN, and 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 Alkyl groups, SF5;

[0117] R 7 Selected from H, halogens, 5-membered heteroaryl groups, C 1-2 Alkyl, C 1-2 Alkoxy, C 2-4 alkenyl, C 2-4 alkynyl, halogenated C 1-2 Alkyl, -C(O)C 1-2 Alkyl, =O, =CH2, =CF2, =C(CH3)2, wherein the alkyl, alkenyl, alkynyl, alkoxy, or heteroaryl groups are optionally further selected from 1-3 halogens, deuterium, CN, OH, C 1-2Alkyl, Halogenated C 1-2 Alkyl, deuterated C 1-2 Alkyl, C 1-2 Alkoxy, halogenated C 1-2 Alkoxy, deuterated C 1-2 Alkoxy group substitution;

[0118] The definitions of other functional groups are consistent with any of the technical solutions mentioned above.

[0119] The fifth technical solution of the present invention is the compound of formula (I) described in the present invention, its stereoisomers, and pharmaceutically acceptable salts, wherein,

[0120] Selected from

[0121]

[0122] This indicates the site linked to L1, where L1 is a bond. This indicates the site connected to ring B.

[0123] Selected from

[0124]

[0125] * indicates the linking site between loop B and L2. When L2 is a bond, * indicates the linking site between loop B and C. ** indicates the linking site between loop B and L1. When L1 is a bond, ** indicates the linking site between loop B and loop A.

[0126] Selected from

[0127]

[0128] or Selected from

[0129] The definitions of other functional groups are consistent with any of the technical solutions mentioned above.

[0130] The sixth technical solution of the present invention is that the compound of formula (I) of the present invention, its stereoisomers and pharmaceutically acceptable salts, have the structures of formulas (II) and (III):

[0131]

[0132] in,

[0133] Selected from

[0134] R 7 Selected from H, halogen, OH, NH2, CN, =O, =CH2, =CF2, =C(CH3)2, methyl, ethyl, trifluoromethyl, cyclopropyl;

[0135] R 6 Selected from halogens;

[0136] R 1 Selected from

[0137] R 2 Selected from H, halogens;

[0138] R 4 Selected from H;

[0139] Or R 2 R 4 Together with the atoms they are attached to, they form 5-7 membered carbon rings;

[0140] X is selected from CH or N;

[0141] Selected from

[0142] The definitions of other functional groups are consistent with any of the technical solutions mentioned above.

[0143] The compounds of formulas (I-1), (I-2), (I-3), (I-4), (I-5), (I-6), (I-7), (I-8), (I-9), (I-10), (I-11), (I-12), (I-13), (I-14), (I-15), (I-16), (II), and (III) of this invention, their stereoisomers, and pharmaceutically acceptable salts, are selected from, but not limited to, those listed in Table 1:

[0144] Table 1

[0145]

[0146]

[0147]

[0148]

[0149]

[0150]

[0151]

[0152]

[0153] Secondly, the present invention also provides a pharmaceutical composition comprising the compound described in any of the foregoing technical solutions, its stereoisomer, a pharmaceutically acceptable salt, and a pharmaceutically acceptable carrier and / or excipient.

[0154] Furthermore, the pharmaceutical composition or pharmaceutical preparation comprises 1-1500 mg of the compound described in any of the foregoing technical solutions, its stereoisomer, a pharmaceutically acceptable salt, and a pharmaceutically acceptable carrier and / or excipient.

[0155] Furthermore, the present invention also provides the use of the compounds described in any of the foregoing embodiments, their stereoisomers, pharmaceutically acceptable salts, or pharmaceutical compositions in the preparation of medicaments for treating / preventing Myosin II-mediated diseases. Furthermore, Myosin II-mediated diseases include, but are not limited to, muscular dystrophy.

[0156] The present invention also provides a method for treating a disease in mammals, the method comprising administering to a subject a therapeutically effective amount of the compound described in any of the foregoing technical solutions, its stereoisomers, pharmaceutically acceptable salts, and pharmaceutically acceptable carriers and / or excipients, the therapeutically effective amount preferably being 1-1500 mg, wherein the disease is preferably muscular dystrophy.

[0157] The present invention also provides a method for treating diseases in mammals, comprising administering to the mammal a therapeutically effective amount of the compound of the present invention or its stereoisomers, pharmaceutically acceptable salts, or pharmaceutical compositions thereof. In some embodiments, the mammals described in the present invention include humans.

[0158] The term "effective amount" or "therapeutic effective amount" as used in this application means that administering a sufficient amount of the compound disclosed in this application will 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 mitigation of the signs, symptoms, or causes of the disease, or any other desired alteration of the biological system. For example, an "effective amount" for therapeutic use is the amount of a composition comprising the compound disclosed in this application required to provide a clinically significant reduction in disease symptoms. Examples of therapeutically effective doses 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. ;

[0159] This invention relates to a pharmaceutical composition or pharmaceutical formulation comprising a therapeutically effective amount of the compound or its stereoisomer, a pharmaceutically acceptable salt, and a carrier and / or excipients. The pharmaceutical composition may be in unit dosage form (the amount of the active ingredient in a unit dosage form is also referred to as a "dosage 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 mg, etc. The compounds of the present invention or their stereoisomers, or pharmaceutically acceptable salts thereof, in the following amounts: g, 190 mg, 200 mg, 210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 275 mg, 300 mg, 325 mg, 350 mg, 375 mg, 400 mg, 425 mg, 450 mg, 475 mg, 500 mg, 525 mg, 550 mg, 575 mg, 600 mg, 625 mg, 650 mg, 675 mg, 700 mg, 725 mg, 750 mg, 775 mg, 800 mg, 850 mg, 900 mg, 950 mg, 1000 mg, 1100 mg, 1200 mg, 1300 mg, 1400 mg, 1500 mg.

[0160] A method for treating a disease in mammals, the method comprising administering to a subject a therapeutically effective amount of the compound of the present invention, its stereoisomer, or a pharmaceutically acceptable salt, and a pharmaceutically acceptable carrier and / or excipient, the therapeutically effective amount preferably being 1-1500 mg, the disease preferably being muscular dystrophy.

[0161] A method for treating a disease in mammals. The method comprises administering a pharmaceutical compound of the invention, its stereoisomers, a pharmaceutically acceptable salt, and a pharmaceutically acceptable carrier and / or excipient to a subject at a daily dose of 1-1500 mg / day. The daily dose may be a single dose or multiple 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... -1000mg / day, 200-1000mg / day, 25-800mg / day, 50-800mg / day, 100-800mg / day, 200-800mg / day, 25-400mg / day, 50-400mg / day, 100-400mg / day, 200-400mg / day. In some embodiments, the daily dose includes, but is not limited to, 1mg / day, 5mg / day, 10mg / day, 20mg / day, 25mg / day, 50mg / day, 75mg / day, 100mg / day, 125mg / day, 150mg / day, 200mg / day, 400mg / day, 600mg / day, 800mg / day, 1000mg / day, 1200mg / day, 1400mg / day, and 1500mg / day.

[0162] This invention relates to a kit that may include a single-dose or multi-dose composition comprising a compound of the present invention or its stereoisomers, or a pharmaceutically acceptable salt thereof, wherein the amount of the compound of the present invention or its stereoisomers, or the pharmaceutically acceptable salt thereof is the same as the amount in the aforementioned pharmaceutical composition.

[0163] In this invention, the amount of the compound or its stereoisomer, or pharmaceutically acceptable salt thereof, is converted in each case as a free base.

[0164] "Product specification" refers to the weight of the active pharmaceutical ingredient contained in each vial, tablet, or other unit of preparation.

[0165] Synthetic route

[0166] Those skilled in the art can prepare the compounds of this invention using known organic synthesis techniques, with starting materials being commercially available chemicals and / or compounds described in chemical literature. "Commercially available chemicals" are obtained from legitimate commercial sources, and suppliers include: Titan Technology, Energie Chemicals, Shanghai Demo, Chengdu Kelon Chemical, Shaoyuan Chemical Technology, Nanjing Yaoshi, WuXi AppTec, and Bailingwei Technology, among others.

[0167] Indexes of known chemical substances prepared by the American Chemical Society's Chemical Abstracts Service can selectively identify specific and similar reactants. These indexes are available in most public and university libraries, as well as online. Known but not commercially available chemicals in the catalogue can optionally be prepared by custom chemical synthesis plants, many of which offer custom synthesis services to standard chemical supply plants (such as those listed above).

[0168] the term

[0169] Unless otherwise specified in this invention, the terminology used in this invention has the following meanings:

[0170] The carbon, hydrogen, oxygen, sulfur, nitrogen, or halogen involved in the groups and compounds described in this invention all include their isotopes, and the carbon, hydrogen, oxygen, sulfur, nitrogen, or halogen involved in the groups and compounds described in this invention may optionally be further replaced by one or more of their corresponding isotopes, wherein the isotopes of carbon include 12 C 13 C and 14 C, the isotopes of hydrogen include protium (H), deuterium (also known as heavy hydrogen), and tritium (T, also known as superheavy hydrogen), and the isotopes of oxygen include 16 O、 17 O and 18 O, isotopes of sulfur include 32 S, 33 S, 34 S and 36 S, nitrogen isotopes include 14 N and 15 N, an isotope of fluorine 19 F, isotopes of chlorine include 35 Cl and 37 Cl, isotopes of bromine include 79 Br and 81 Br.

[0171] In this article, "halogen" refers to F, Cl, Br, I, or their isotopes.

[0172] "Halogenation" or "halogen substitution" refers to substitution by one or more halogen substituents selected from F, Cl, Br, I, 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 substituted in the substituted group. Unless otherwise specified, the number of halogen substituents can be any integer between 1 and this upper limit. When the number of halogen substituents is greater than 1, the substitution can be by the same or different halogens. Common cases include 1-5 halogen substitutions, 1-3 halogen substitutions, 1-2 halogen substitutions, and 1 halogen substitution.

[0173] "Deuterium" refers to the hydrogen (H) isotope deuterium, which is synonymous with "D".

[0174] "Deuteration" refers to the substitution of hydrogen atoms on alkyl, cycloalkyl, alkylene, aryl, heteroaryl, mercapto, heterocycloalkyl, alkenyl, alkynyl, etc. groups by at least one deuterium atom. The upper limit of the number of deuterations is equal to the sum of the number of hydrogen atoms that can be substituted in the substituted group. Unless otherwise specified, the number of deuterations is any integer between 1 and this upper limit, such as 1-20 deuterium atoms, 1-10 deuterium atoms, 1-6 deuterium atoms, 1-3 deuterium atoms, 1-2 deuterium atoms, or 1 deuterium atom.

[0175] “C x-y A "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.

[0176] "Alkyl" refers to a monovalent straight-chain or branched saturated aliphatic hydrocarbon group. It is typically an alkyl group with 1 to 20 carbon atoms, or 1 to 8 carbon atoms, or 1 to 6 carbon atoms, or 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, etc., and the alkyl group may be further substituted with substituents.

[0177] "Alkylene" refers to divalent straight-chain and branched saturated alkyl groups. Examples of alkylene groups include, but are not limited to, methylene, ethylene, etc.

[0178] "Halogenated alkyl" refers to an alkyl group in which one or more hydrogen atoms 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 substituted in the alkyl group. Unless otherwise specified, the number of halogen substituents is any integer between 1 and this upper limit. Typically, alkyl groups are 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.

[0179] "Alkoxy" or "alkyloxy" refers to -O-alkyl. For example, -OC 1-8 Alkyl, -OC 1-6 Alkyl, -OC 1-4 Alkyl or -OC 1-2 Alkyl groups. Specific, non-limiting examples include methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, sec-butoxy, tert-butoxy, n-pentoxy, n-hexoxy, cyclopropoxy, and cyclobutoxy, etc.; the alkoxy groups may optionally be substituted with substituents.

[0180] "Haloalkoxy" refers to -O-haloalkyl. 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 groups; the upper limit of the number of halogen substituents is equal to the sum of the number of hydrogens that can be replaced in the substituted group. Unless otherwise specified, the number of halogen substituents is any integer between 1 and the upper limit, preferably 1-5 halogen substituents, 1-3 halogen substituents, 1-2 halogen substituents, or 1 halogen substituent. When the number of halogen substituents is greater than 1, the same or different halogens can be used for substitution. Non-limiting examples include monofluoromethoxy, difluoromethoxy, trifluoromethoxy, difluoroethyloxy, etc.

[0181] "Alkenyl" refers to a straight-chain or branched 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 even further such as 2 to 4 carbon atoms. Examples 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-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 optionally be further substituted with substituents.

[0182] "Alkenyl" refers to a straight-chain or branched divalent unsaturated 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 even further such as 2 to 4 carbon atoms. Non-limiting examples include ethynylene groups, which may optionally be substituted with substituents.

[0183] "Alynyl" 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-hepynyl, 3-hepynyl, 4-hepynyl, 3-octyynyl, 3-nonynyl, and 4-decynyl; the alkynyl group may optionally be substituted with substituents.

[0184] "Imyynyl" refers to a straight-chain or branched divalent unsaturated hydrocarbon group containing a 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. Non-limiting embodiments include ethynyl, propynyl, and butynyl, wherein the ethynyl group may optionally be substituted with substituents.

[0185] "Cycloalkyl" refers to a saturated or partially unsaturated, non-aromatic carbocyclic hydrocarbon group that does not contain cyclic heteroatoms. Cycloalkyl groups can be monocyclic, bicyclic, or polycyclic. Bicyclic or polycyclic groups can be fused, spirocyclic, bridged, or combinations thereof. A bicyclic or polycyclic group may include one or more aromatic rings, but the ring system as a whole is not aromatic. The linkage site can be on an aromatic ring or a non-aromatic ring. Typically, cycloalkyl groups contain 3 to 20 carbon atoms, more commonly 3 to 8 carbon atoms, and even more commonly 3 to 6 carbon atoms. When it is a monocyclic cycloalkyl group, it contains 3 to 15 carbon atoms, or 3 to 10 carbon atoms, or 3 to 8 carbon atoms, or 3 to 6 carbon atoms. When it is a bicyclic or polycyclic cycloalkyl group, it contains 5 to 12 carbon atoms, or 5 to 11 carbon atoms, or 6 to 10 carbon atoms. Non-limiting examples include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, butenyl, cyclopentenyl, and cyclohexenyl. In addition, cycloalkyl groups may optionally be substituted with substituents.

[0186] "Cycloalkylene" refers to the divalent group of a cycloalkyl group.

[0187] "Aryl" refers to an aromatic carbon ring that does not contain heteroatoms, including monocyclic aryl and fused-ring aryl. It typically contains 6 to 14 carbon atoms, and more commonly 6 to 10 carbon atoms. Non-limiting examples include phenyl, naphthyl, anthraceneyl, and phenanthreneyl groups, and the aryl group may optionally be substituted with substituents.

[0188] "Carbocyclic" or "carbocyclic group" refers to a saturated, partially unsaturated, or aromatic carbon ring, including aryl and cycloalkyl groups. The carbon ring can be monocyclic, bicyclic, or polycyclic, including bridged rings, fused rings, and spirocyclic rings, as well as combinations thereof. Carbon rings typically have 3 to 12 carbon atoms, or 3 to 10 carbon atoms, or 3 to 6 carbon atoms. In non-limiting embodiments, monocyclic carbon rings include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, or phenyl, etc., and bicyclic bridged rings include... Etc., double-ring parallel rings include etc., double-ring spiral rings include In addition, the carbon ring can be optionally replaced by substituents.

[0189] "Heterocyclic alkyl" refers to a non-aromatic ring containing 1, 2, 3, or 4 heteroatoms selected from N, S, O, and Si, either saturated or partially unsaturated. Heterocyclic alkyl groups can be monocyclic, bicyclic, or polycyclic. Bicyclic or polycyclic rings can be bridged, fused, spirocyclic, or combinations thereof. A bicyclic or polycyclic ring may include one or more aromatic or heteroaromatic rings, but the ring system as a whole is not aromatic. Linking sites can be on aromatic or non-aromatic rings. Typically, heterocyclic alkyl groups are 3 to 20-membered rings. When monocyclic, they are typically 3 to 15-membered, or 3-10-membered, or 3-8-membered, or 3-6-membered rings. When bicyclic or polycyclic heterocyclic alkyl groups, they are typically 5-12-membered, or 5-11-membered, or 6-9-membered rings. The heteroatoms N and S include their oxidation states. Non-limiting examples of heterocyclic alkyl groups include azirrobutyl, morpholino, piperazinyl, piperidinyl, tetrahydropyranyl, oxacyclobutyl, pyranyl, azirropentenyl, azirrohexenyl, oxacyclopentenyl, oxacyclohexenyl, etc., and the heterocyclic alkyl groups may optionally be substituted with substituents.

[0190] Unless otherwise specified, "heteroaromatic ring" or "heteroaryl" refers to an aromatic ring containing 1 to 4 heteroatoms or oxo groups selected from N, O, S, and their oxidation states. It can be monocyclic, bicyclic, or polycyclic. Bicyclic or polycyclic rings can be bridged rings, fused rings, spirocyclic rings, and combinations thereof. When bicyclic or polycyclic, it can be a fusion of a heteroaryl group and an aryl group, or a fusion of two heteroaryl groups, wherein either the heteroaryl group or the aryl group can be a linking site. Non-limiting examples include furanyl, thiophene, pyrrole, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, pyridinyl, pyrimidinyl, pyridazinyl, indoleyl, purine, etc. The heteroaryl group can be optionally substituted with a substituent.

[0191] "Heterocycle" or "heterocyclic group" refers to a saturated or unsaturated, aromatic or non-aromatic ring containing one to four heteroatoms selected from N, O, or S and their oxidation states. It includes heteroaryl and heterocyclic alkyl groups. Heterocycles include monocyclic heterocycles, bicyclic bridged heterocycles, bicyclic fused heterocycles, and bicyclic spirocyclic heterocycles, or combinations thereof. They are typically 3- to 12-membered heterocycles, 5- to 12-membered heterocycles, or 5- to 7-membered heterocycles. Heterocyclic groups can be attached to heteroatoms or carbon atoms. Non-limiting examples include epoxyethyl, azirropropyl, oxacyclobutyl, azirrobutyl, 1,3-dioxopentyl, 1,4-dioxopentyl, 1,3-dioxohexyl, piperazine, azirroheptyl, pyridinyl, furanyl, thiophene, pyranyl, N-alkylpyrroleyl, pyrimidinyl, pyrazinyl, pyrazolyl, pyridazinyl, imidazoleyl, piperidinyl, piperinyl, morpholinyl, thiomorpholinyl, and 1,3-dithiayl. Dihydrofuranyl, dihydropyranyl, dithiapentylcycloyl, tetrahydrofuranyl, tetrahydropyrroleyl, tetrahydroimidazoyl, oxazolyl, dihydrooxazolyl, tetrahydrooxazolyl, tetrahydrothiazoyl, tetrahydropyranyl, benzimidazolyl, benzopyridyl, pyrrolopyridyl, benzodihydrofuranyl, azabicyclo[3.2.1]octyl, azabicyclo[5.2.0]nonyl, oxatricyclo[5.3.1.1]dodecyl, azaadamantyl and oxaspiro[3.3]heptyl, In addition, heterocyclic rings can be optionally substituted by substituents.

[0192] "Hypo-heterocyclic group" refers to a divalent heterocyclic group that is substituted or unsubstituted, saturated or unsaturated, aromatic or non-aromatic. Non-limiting examples include... wait.

[0193] A "spirocyclic ring" is a polycyclic group that shares a single carbon atom (called a spiro atom) between rings. It can contain 0 or more double or triple bonds and 0 to 5 heteroatoms selected from N, O, S, P, Si, and their oxidation states. Spirocyclic rings are typically 6 to 14-membered, 6 to 12-membered, or 6 to 10-membered. Common spirocyclic rings include tri-spirotri- (representing a three-membered ring followed by a spirotri-membered ring), tri-spirotetra-, tri-spiropenta-, tri-spirohexa-, tetra-spirotetra-, tetra-spiropenta-, tetra-spirohexa-, tetra-spiropenta-, tetra-spirohexa-, penta-spiropenta-, or penta-spirohexa-. Non-limiting examples of spirocyclic rings include...

[0194]

[0195] The spiroring can be optionally replaced by a substituent.

[0196] "Fused rings" or "fused rings" refer to polycyclic groups where rings share two adjacent ring atoms and a chemical bond. They can contain one or more double or triple bonds and can contain 0 to 5 heteroatoms selected from N, S, O, P, Si, and their oxidation states. Fused rings are typically 5 to 20-membered, 5 to 14-membered, 5 to 12-membered, or 5 to 10-membered rings. Common fused ring types include trifused tetracyclic rings (representing fused rings formed by a three-membered and a four-membered ring; according to IUPC nomenclature, this could be a fused ring with either a three-membered or a four-membered ring as the base ring, and the same applies below), trifused pentacyclic rings, trifused hexacyclic rings, tetrafused tetracyclic rings, tetrafused pentacyclic rings, tetrafused hexacyclic rings, pentafused pentacyclic rings, pentafused hexacyclic rings, and hexafused hexacyclic rings. Non-limiting examples of fused rings include purines, quinolines, isoquinolines, benzopyrans, benzofurans, benzothiophenes, etc. The cyclic ring can be aromatic or non-aromatic, and optionally substituted with substituents.

[0197] A "bridged ring" refers to two rings sharing two non-adjacent ring atoms and may contain one or more double or triple bonds. Bridged rings can contain 0 to 5 heteroatoms selected from N, S, O, P, Si, and their oxidation states. Typically, bridged rings have 5 to 20, 5 to 14, 5 to 12, or 5 to 10 ring atoms. Non-limiting examples of bridged rings include adamantane, etc.

[0198]

[0199] Unless otherwise specified, "substitution" or "substituent" refers to any substitution occurring at a position permitted by chemical theory, with the number of substituents conforming to the rules of chemical bonding. Exemplary substituents include, but are not limited to: C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 3-8 Heteroalkyl, C 5-12 Aryl, 5-12 heteroaryl, hydroxyl, C 1-6 Alkoxy, C 5-12 aryloxy groups, thiol groups, C 1-6 Alkylthio, cyano, halogen, C 1-6 alkylthiocarbonyl, C 1-6 Alkyl carbamoyl, N-carbamoyl, nitro, silyl, sulfinyl, sulfonyl, sulfoxide, halogenated C 1-6 Alkyl, Halogenated C 1-6 Alkyl, 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 1-6Alkyl), -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 group, -SO2NH2, -SO2C 1-6 Alkyl groups, etc.

[0200] "Pyridazinone" refers to pyridazine or pyridazin groups containing =O, such as "Pyridone" refers to pyridine or pyridinyl groups containing the =O group, such as... "Pyrazinone" refers to a pyrazine or pyrazin group containing the =O radical, such as... "Pyrimidine ketone" refers to a pyrimidine or pyrimidine group containing the =O group, such as...

[0201] "Optional" or "optionally" means that the event or environment described below may but does not have to occur, and the description includes the possibility or possibility that the event or environment may or may not occur. For example, "optionally substituted F alkyl" means that the alkyl group may but does not have to be substituted with F, and the description includes the case where the alkyl group is substituted with F and the case where the alkyl group is not substituted with F.

[0202] "Pharmaceutically acceptable salt" means that the compound of the present invention retains the bioavailability and properties of a free acid or a free base, wherein the free acid is obtained by reacting with a non-toxic inorganic or organic base, and the free base is obtained by reacting with a non-toxic inorganic or organic acid.

[0203] "Pharmaceutical composition" means one or more of the compounds described herein or their stereoisomers, pharmaceutically acceptable salts, and mixtures with other components, wherein the other components contain physiologically / pharmaceuticalally acceptable carriers and / or excipients.

[0204] "Carrier" refers to a system that does not cause significant stimulation to the organism and does not eliminate the biological activity and properties of the given 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.

[0205] "Excipients" refer to substances that are not therapeutic agents themselves, but are used as diluents, excipients, binders, and / or mediators to be added to pharmaceutical compositions to improve their disposal or storage properties or to allow or facilitate the formation of unit dosage forms of compounds or pharmaceutical compositions for administration. As known to those skilled in the art, excipients can provide 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 carboxymethyl cellulose, ethyl cellulose, cellulose acetate, hydroxypropyl methyl cellulose, hydroxypropyl cellulose, microcrystalline cellulose and croscarmellose (e.g. croscarmellose sodium); (4) tragacanth gum powder; (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 and corn oil. (10) Soybean oil; (11) Diols, such as propylene glycol; (12) Polyols, such as glycerol, sorbitol, mannitol and polyethylene glycol; (13) Esters, such as ethyl oleate and ethyl laurate; (14) Agar; (15) Buffers, such as magnesium hydroxide and aluminum hydroxide; (16) Alginate; (17) Atherless water; (18) Isotonic saline; (19) Ringer's solution; (20) Ethanol; (21) pH buffer solution; (22) Polyesters, polycarbonates and / or polyanhydrides; and (23) Other non-toxic compatible substances used in pharmaceutical preparations.

[0206] "Stereoisomers" refer to isomers that have the same order of interconnection of atoms or groups of atoms in a molecule but different spatial arrangements. All such compounds of this invention, including cis-trans isomers, optical isomers, and racemic mixtures and other mixtures thereof, such as cis and trans isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers, (D)- isomers, (L)- isomers, and racemic mixtures and other mixtures thereof, such as mixtures enriched with enantiomers or diastereomers, are all within the scope of this invention. Additional asymmetric carbon atoms may be present in the substituents of the compounds of this invention. All such isomers and mixtures thereof are included within the scope of this invention. In some embodiments, the preferred compounds are those isomers that exhibit superior biological activity. Purified or partially purified isomers and stereoisomers, or racemic mixtures or diastereomer mixtures of the compounds of this invention are also included within the scope of this invention. The purification and separation of such substances can be achieved using standard techniques known in the art.

[0207] The compounds of this invention also include their tautomers, which are interconvertible via a reversible chemical reaction known as tautomerization, typically resulting from the associated migration of hydrogen atoms and π bonds (double or triple bonds), transforming one functional group into another. Examples include paired compounds such as aldehydes / ketones–enols, imines–enamines. For instance, when this invention describes the left-hand compound with a pyrimidine ring substituted with OH, the right-hand tautomer compound is also included, for example... Detailed Implementation

[0208] The present invention will be described in detail below through embodiments. Unless otherwise specified, experimental methods under conventional conditions were used in the embodiments. The embodiments are provided to better illustrate the present invention, but should not be construed as limiting the invention to the examples given. Non-essential improvements and adjustments made to the implementation schemes by those skilled in the art based on the above description are still within the scope of protection of the present invention.

[0209] The structure of the compound was determined by nuclear magnetic resonance (NMR) and / or mass spectrometry (MS). NMR shifts (δ) were expressed in 10⁻¹⁰ increments. -6 The unit (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).

[0210] MS determination was performed using (Agilent 6120B (ESI) and Agilent 6120B (APCI));

[0211] The HPLC determination was performed using an Agilent 1260DAD high-performance liquid chromatograph (Zorbax SB-C). 18 100×4.6mm, 3.5μM);

[0212] Thin-layer chromatography silica gel plates are Yantai Huanghai HSGF254 or Qingdao GF254. The silica gel plates used in thin-layer chromatography (TLC) are 0.15mm-0.20mm in diameter, and the silica gel plates used for thin-layer chromatography separation and purification are 0.4mm-0.5mm in diameter.

[0213] Column chromatography typically uses Yantai Huanghai silica gel with a mesh size of 200-300 as the carrier.

[0214] Example 1

[0215]

[0216] Step 1: 2,2,2-Trifluoroethanol (5.17 g, 51.7 mmol) was added to a tetrahydrofuran (100 mL) solution. Sodium hydride (2.48 g, 62.04 mmol) was added under ice bath conditions. After stirring at room temperature for 30 minutes, compound 1A (10 g, 51.70 mmol) was added, and the reaction was allowed to proceed at room temperature for 2 hours. After the reaction was complete, the reaction mixture was poured into ice water, and ethyl acetate (200 mL) was added for extraction. The mixture was separated, and the organic phase was washed with water (100 mL × 3). The organic phase was dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel column chromatography to give compound 1B (12.05 g, yield: 90.69%). LC-MS (ESI): m / z = 257.0 [M + H] + .

[0217] Step 2: Compound 1B (11 g, 42.80 mmol), pinacol diboronate (13.05 g, 51.38 mmol), tricyclohexylphosphine (1.20 g, 4.28 mmol), and potassium acetate (10.50 g, 107 mmol) were added to a 1,4-dioxane (100 mL) solution. The mixture was purged with nitrogen three times. Palladium acetate (0.48 g, 2.14 mmol) was then added, followed by another three purgings with nitrogen. The mixture was heated to 100 °C and reacted for 2 hours. After the reaction was complete, the mixture was filtered, and the filtrate was concentrated to obtain compound 1C (9.5 g crude product), which was directly used in the next step of the reaction. LC-MS (ESI): m / z = 223.1 [M+H] + .

[0218] Step 3: Compound 1C (9.5 g, 42.81 mmol), 6-bromopyridazine-3(2H)-one (7.49 g, 42.81 mmol), and potassium phosphate (13.63 g, 64.22 mmol) were added to a 1,4-dioxane (100 mL) solution. The mixture was purged with nitrogen three times. Then, 1,3-bis(2,6-diisopropylphenyl)imidazolium-2-ylylene](3-chloropyridinyl)palladium(II) dichloride (2.92 g, 4.28 mmol) was added, and the mixture was purged with nitrogen three times again. The mixture was heated to 100 °C and reacted for 2 hours. After cooling, the mixture was filtered directly. The filter cake was washed sequentially with 1,4-dioxane (20 mL), dichloromethane (20 mL), and water (20 mL). The filter cake was collected and dried to give compound 1D (3 g, yield: 100%). LC-MS (ESI): m / z = 273.1 [M+H] + .

[0219] Step 4: Compound 1D (800 mg, 2.94 mmol), 3-chloro-5-chloromethylpyridine (0.48 g, 2.94 mmol), and potassium carbonate (1.02 g, 7.35 mmol) were added to a solution of N,N-dimethylformamide (10 mL), and the mixture was heated to 80 °C for 2 hours. After cooling, ethyl acetate (100 mL) and water (100 mL) were added for liquid-liquid extraction. The organic phase was washed with water (100 mL × 3) and dried over anhydrous sodium sulfate. The residue was concentrated and purified by reversed-phase column chromatography to give compound 1 (210 mg, yield: 18%). 1 HNMR(400MHz, CDCl3)δ8.86(d,1H),8.64(d,1H),8.53(d,1H),8.34(d,1H),8.19(d,1H),7. 84-7.83(m,1H),7.07(d,1H),5.40(s,2H),4.87-4.81(m,2H); LC-MS(ESI):m / z=398.1[M+H] + .

[0220] Example 2

[0221]

[0222] Step 1: Compound 2A (2.8 g, 20.0 mmol) was dissolved in tetrahydrofuran (50.0 mL), and diisobutylaluminum hydride (33.3 mL, 50.0 mmol, 1.5 M tetrahydrofuran solution) was slowly added under ice bath conditions. The mixture was slowly heated to room temperature and reacted for 1 h. Water (2.0 mL), 15% sodium hydroxide solution (2.0 mL), and water (5.0 mL) were added successively. The mixture was stirred for 0.5 h, and then anhydrous sodium sulfate was added to dry the mixture. The mixture was filtered, and the filtrate was collected and concentrated. The residue was purified by silica gel column chromatography to obtain compound 2B (1.6 g, yield: 85%). 1 H NMR (400MHz, CDCl3) δ5.41–5.25(m,1H),4.01(d,2H),2.74–2.67(m,4H),2.01-1.93(m,2H).

[0223] Step 2: Compound 2B (1.0 g, 10.0 mmol) was dissolved in dichloromethane (50 mL), and triethylamine (203.0 mg, 20.0 mmol) and methanesulfonyl chloride (1.2 g, 11.0 mmol) were added. The reaction was carried out at room temperature for 2 h. The mixture was quenched with water, extracted with dichloromethane (30 mL × 3), and the organic phase was collected and concentrated to give compound 2C (0.7 g crude product, yield: 39%), which was used directly in the next step of the reaction.

[0224] Step 3: Compound 2C (0.47 g, 2.6 mmol) was dissolved in acetonitrile (20 mL), followed by the addition of potassium carbonate (717.0 mg, 5.2 mmol) and 6-bromo-3-pyridazinone (0.45 g, 2.6 mmol). The mixture was heated to 80 °C and reacted for 4 h. After cooling, the mixture was filtered, the filtrate was collected and concentrated, and the residue was purified by silica gel column chromatography to give compound 2D (0.38 g, yield: 57%). LC-MS (ESI): m / z = 255.0 [M+H] + .

[0225] Step 4: Compound 2D (0.4 g, 1.5 mmol) was dissolved in 1,4-dioxane (20 mL), and [1,1'-bis(diphenylphosphine)ferrocene]palladium(II) dichloride (105.0 mg, 0.15 mmol), potassium acetate (290.0 mg, 3.0 mmol), and pinacol diborate (456.0 mg, 1.8 mmol) were added. The mixture was heated to 90 °C and reacted for 2 h. After cooling to room temperature, compound 1B (0.38 g, 1.5 mmol), [1,1'-bis(diphenylphosphine)ferrocene]palladium(II) dichloride (105.0 mg, 0.15 mmol), potassium carbonate (415.0 mg, 3.0 mmol), and water (2.0 mL) were added and heated to 90 °C and reacted for 2 h. After cooling, the filter cake was washed with ethyl acetate and the filtrate was collected and concentrated. The residue was purified by silica gel column chromatography to give compound 2 (0.06 g, yield: 13%). 1 H NMR (400MHz, DMSO-d6) δ8.87(d,1H),8.58(d,1H),8.12(d,1H),7.08(d,1H),5.38–5.34(m,1H),5.15–5.08( m,2H),4.62(d,2H),2.81–2.77(m,2H),2.68–2.64(m,2H),1.97–1.89(m,2H); LC-MS(ESI):m / z=353.1[M+H] + .

[0226] Example 3

[0227]

[0228] Step 1: Triethyl phosphoroacetate (8.22 g, 36.68 mmol) was dissolved in tetrahydrofuran (50.0 mL). Sodium hydride (1.71 g, 42.80 mmol) was added in portions under ice-water bath conditions. After stirring for 30 minutes, cyclohexanone (3A) (3 g, 30.57 mmol) was added, and the mixture was stirred at room temperature for 1 hour. The reaction was quenched with saturated ammonium chloride solution. The mixture was extracted with ethyl acetate (50 mL × 3). The organic phase was washed with saturated brine (100 mL × 3), dried over anhydrous sodium sulfate, and the concentrated residue was purified by silica gel column chromatography to give compound 3B (2.5 g, yield: 48%). LC-MS (ESI): m / z = 169.1 [M + H] + .

[0229] Step 2: Compound 3B (0.5 g, 3.0 mmol) was dissolved in tetrahydrofuran (20.0 mL), and diisobutylaluminum hydride (9.0 mL, 1.0 M n-hexane solution, 9.0 mmol) was slowly added under ice-water bath conditions. The mixture was slowly heated to room temperature and stirred for 1 h. Water (0.4 mL), 15% sodium hydroxide solution (0.4 mL), and water (1.0 mL) were added sequentially. After stirring for 0.5 h, anhydrous sodium sulfate was added for drying. The mixture was filtered, and the filtrate was collected and concentrated to obtain compound 3C (0.25 g crude product, yield: 66%), which was directly used in the next reaction.

[0230] Step 3: Compound 3C (0.25 g, 1.98 mmol) was dissolved in dichloromethane (10.0 mL), and triethylamine (0.50 g, 4.95 mmol) and p-toluenesulfonyl chloride (0.57 g, 2.97 mmol) were added. The reaction was carried out at room temperature for 4 h. The reaction was quenched with water, and the mixture was extracted with dichloromethane (20 mL × 3). The organic phase was collected and concentrated to give compound 3D (0.32 g crude product, yield: 57%), which was directly used in the next step of the reaction.

[0231] Step 4: Compound 1D (0.20 g, 0.73 mmol) was dissolved in N,N-dimethylformamide (10 mL), followed by the addition of cesium carbonate (0.36 g, 1.10 mmol) and compound 3D (0.23 g, 0.8 mmol). The mixture was heated to 80 °C and stirred for 3 h. After cooling, the reaction solution was diluted with ethyl acetate (30 mL), washed with saturated brine (30 mL × 2), and the organic phase was dried over anhydrous sodium sulfate. The residue was filtered and concentrated, and then purified by preparative HPLC to obtain compound 3 (60 mg, yield: 21%). 1H NMR(400MHz,DMSO-d6)δ8.88(d,1H),8.57(d,1H),8.11(d,1H),7.08(d,1H),5.32-5.28(m,1H),5.15-5.07(m,2H),4.26-4.21(m, 2H),2.43-2.37(m,2H),3.03-1.98(m,2H),1.85-1.79(m,2H),1.57-1.51(m,2H),1.45-1.39(m,2H); LCMS(ESI):m / z=381.2[M+H] + .

[0232] Example 4

[0233]

[0234] Step 1: Compound 4A (0.7 g, 5.22 mmol) was dissolved in dichloromethane (12 mL), and thionyl chloride (1.24 g, 10.44 mmol) was added. The mixture was stirred at room temperature for 2 hours. The solution was then concentrated under reduced pressure to obtain compound 4B (0.8 g crude product, yield: 100%), which was used directly in the next step of the reaction. 1 HNMR (400MHz, DMSO-d6) δ7.24(dd,1H),7.15(t,1H),7.07(dd,1H),4.72(s,2H),3.12(s,4H).

[0235] Step 2: Compound 1D (0.2 g, 0.73 mmol), compound 4B (0.16 g, 1.05 mmol), and potassium carbonate (1.0 g, 2.17 mmol) were dissolved in N,N-dimethylformamide (12 mL), and the mixture was heated to 80 °C and stirred for 3 hours. After cooling, water (10 mL) and ethyl acetate (15 mL × 3) were added for extraction three times. The organic phase was collected, concentrated, and the residue was sent for preparative HPLC purification to obtain compound 4 (130 mg, yield: 45.8%). 1 H NMR(400MHz,DMSO-d6)δ8.89(d,1H),8.57(d,1H),8.13(d,1H),7.26(dd,1H),7.15-7.09(m, 2H),7.05(d,1H),5.29(s,2H),5.16-5.07(m,2H),3.09(s,4H); LC-MS(ESI):m / z=389.1[M+H] + .

[0236] Example 5

[0237]

[0238] Step 1: 2-Cyclobutylethanol (5A) (4.0 g, 40.0 mmol) was dissolved in dichloromethane (50 mL), and triethylamine (810 mg, 80.0 mmol) and methanesulfonyl chloride (4.6 g, 40.0 mmol) were added. The mixture was stirred at room temperature for 2 h. The solution was quenched with water and extracted with dichloromethane (30 mL × 3). The organic phase was collected, dried, and concentrated to give compound 5B (7.8 g crude product, yield: 100%), which was used directly in the next step of the reaction.

[0239] Step 2: Compound 1D (200 mg, 0.73 mmol), compound 5B (0.16 g, 0.88 mmol), and cesium carbonate (0.71 g, 2.19 mmol) were added to N,N-dimethylformamide (20 mL). After reacting at 80 °C for 2 hours, ethyl acetate (100 mL) and water (100 mL) were added for separation. The organic phase was washed with water (100 mL × 3) and dried and concentrated. The residue was purified by reversed-phase column chromatography to obtain compound 5 (200 mg, yield: 77.32%). 1 H NMR(400MHz,MeOD)δ8.92(d,1H),8.40(d,1H),8.27(d,1H),7.07(d,1H),4.02-4.95(m,2H),4.22-4.18(m,2H),2.44-2 .32(m,1H),2.14–2.02(m,2H),1.99-1.93(m,2H),1.92–1.78(m,2H),1.75–1.63(m,2H); LC-MS(ESI):m / z=355.1[M+H] + .

[0240] Example 6

[0241]

[0242] Step 1: 2,2-Difluoroethanol-1-ol (1A) (19.09 g, 232.59 mmol) was dissolved in tetrahydrofuran (80 mL). Sodium hydride (7.44 g, 10 mmol) was slowly added in portions under an ice-water bath. The mixture was stirred at room temperature for 0.5 hours. Then, 2-bromo-5-chloropyrazine (30 g, 155.10 mmol) was dissolved in tetrahydrofuran (80 mL) and added dropwise to the above system under an ice-water bath. The mixture was stirred at room temperature for another 2 hours. The mixture was diluted with water and extracted three times with ethyl acetate. The organic phases were combined and concentrated. The residue was purified by silica gel column chromatography to give compound 6A (29 g, yield: 78%). LC-MS (ESI): m / z = 239.3 [M+H] + .

[0243] Step 2: Compound 6A (15 g, 62.76 mmol), potassium acetate (15.40 g, 156.9 mmol), pinacol diboronate (20.72 g, 81.59 mmol), and chloro(2-dicyclohexylphosphino-2',6'-dimethoxy-1,1'-biphenyl)(2'-amino-1,1'-biphenyl-2-yl)palladium(II) (Sphos Pd G2) (4.52 g, 6.28 mmol) were dissolved in 1,4-dioxane (80 mL). The solution was then purged three times with nitrogen and heated to 100 °C with stirring for 3 hours under a nitrogen atmosphere. The mixture was filtered while hot, and the filter cake was washed four times with ethyl acetate and dichloromethane. The filtrate was collected and concentrated to give compound 6B (12 g crude product, yield: 93%). LC-MS (ESI): m / z = 205.1 [M+H] + .

[0244] Step 3: Compound 6B (13 g, 63.74 mmol), 6-bromo-2,3-dihydropyridazin-3-one (13.38 g, 76.49 mmol), tripotassium phosphate (17.59 g, 82.86 mmol), and (1,3-bis(2,6-diisopropylphenyl)imidazolyl)(3-chloropyridinyl)palladium(II) dichloride (PEPPSI(TM)-SIPR catalyst) (4.34 g, 6.37 mmol) were dissolved in a mixed solvent of 1,4-dioxane (120 mL) and water (40 mL). The solution was then purged three times with nitrogen and heated to 90 °C under a nitrogen atmosphere with stirring for 3 hours. After cooling, the solution was filtered directly. The filter cake was repeatedly washed with ethyl acetate and water. The filtrate was collected and extracted three times with ethyl acetate. The organic phase was collected and concentrated. The residue was purified by silica gel column chromatography to give compound 6C (8.3 g, yield: 51%). LC-MS (ESI): m / z = 255.1 [M+H] + .

[0245] Step 4: Compound 6C (0.21 g, 0.81 mmol), compound 4B (0.19 g, 1.22 mmol), and potassium carbonate (0.28 g, 2.03 mmol) were dissolved in N,N-dimethylformamide (12 mL), and the mixture was heated to 70 °C and stirred for 3 hours. After cooling, the reaction solution was diluted with water, and then extracted three times with ethyl acetate. The organic phases were combined and concentrated, and the residue was sent for preparative HPLC purification to obtain compound 6 (30 mg, yield: 10%). 1H NMR(400MHz,DMSO-d6)δ8.88(d,1H),8.50(d,1H),8.13(d,1H),7.26(d,1H),7.15–7.09(m,2H),7.05( d,1H),6.61-6.29(m,1H),5.29(s,2H),4.77-4.63(m,2H),3.09(s,4H); LC-MS(ESI):m / z=371.2[M+H] + .

[0246] Example 7

[0247]

[0248] Step 1: Compound 1D (500 mg, 1.84 mmol), 4-chlorobenzyl chloride (0.35 g, 2.19 mmol), and potassium carbonate (0.64 g, 4.63 mmol) were added to N,N-dimethylformamide (20 mL), and the mixture was heated to 80 °C for 2 hours. After cooling, ethyl acetate (100 mL) and water (100 mL) were added for separation. The organic phase was washed with water (100 mL × 3), dried over anhydrous sodium sulfate, and concentrated. The residue was purified by reversed-phase column chromatography to give compound 7 (650 mg, yield: 89.04%). 1 H NMR(400MHz,CD3OD)δ8.91(d,1H),8.39(d,1H),8.28(d,1H),7.47–7.40(m,2H),7.37–7 .31(m,2H),7.11(d,1H),5.40(s,2H),5.01-4.95(m,2H); LC-MS(ESI):m / z=397.0[M+H] + .

[0249] Example 8

[0250]

[0251] Step 1: Add 400 mL of tetrahydrofuran to a flask, and add 312 mL of 624.21 mmol of 2 M tetrahydrofuran solution at -78 °C. Dilute 50 g of 2-cyclohexen-1-one (505.1 mmol) with 100 mL of tetrahydrofuran and add it dropwise to the above system. Maintain the temperature and continue the reaction for 1 hour. Then dilute 104.3 g of ethyl bromofuran (100 mL) with 100 mL of tetrahydrofuran and add it dropwise to the system. React at -78 °C for 1 hour, then gradually restore the temperature to room temperature and react overnight. The reaction was quenched by adding 100 mL of saturated ammonium chloride solution dropwise to the system at -78 °C. Extraction was then performed by adding 500 mL of water and 200 mL × 3 of ethyl acetate. The organic phase was collected, washed, dried, and concentrated. The residue was purified by column chromatography (petroleum ether:ethyl acetate (v:v) = 5:1) to give compound 8A (23 g, yield: 24.26%). LCMS (ESI): m / z = 183.2 [M+H] + .

[0252] Step 2: Compound 8A (17 g, 93.29 mmol) was added to a flask, followed by methanol (250 mL). Then, under ice bath conditions, 30% hydrogen peroxide (64 mL, 626.64 mmol) was added, followed by sodium hydroxide solution (1.87 g, 46.65 mmol, dissolved in 13 mL of water). The mixture was gradually heated to room temperature and reacted for 16 h. The reaction was quenched by adding saturated sodium thiosulfate solution under ice bath conditions. The mixture was extracted with ethyl acetate, and the organic phase was collected, washed, and concentrated. The residue was purified by column chromatography (petroleum ether:ethyl acetate (v:v) = 10:1) to give compound 8B (8.2 g, yield: 44.34%). LCMS (ESI): m / z = 199.1 [M+H] + .

[0253] Step 3: Diphenyldiselenoether (19.37 g, 62.05 mmol) was added to a flask, followed by ethanol (290 mL). Sodium borohydride (4.7 g, 124.11 mmol) was added in portions at room temperature, and the reaction was allowed to proceed for 10 minutes. Acetic acid (0.45 mL) was then added, followed by the dissolution of compound 8B (8.2 g, 41.37 mmol) in ethanol (20 mL), which was then added to the above system. The reaction was continued at room temperature for 3 hours. Water was added to the system, and the mixture was extracted with dichloromethane. The organic phase was collected, washed, dried, and concentrated. The residue was purified by column chromatography (petroleum ether:ethyl acetate (v:v) = 1:1) to give compound 8C (7.1 g, yield: 85.71%). LCMS (ESI): m / z = 201.2 [M+H] + .

[0254] Step 4: Compound 8C (8.7 g, 43.45 mmol) was added to a flask, followed by ethanol (160 mL) and hydrazine hydrate (17 mL, 349.78 mmol). The mixture was then reacted at 85 °C for 1 hour. After cooling, the mixture was directly concentrated, and the residue was purified by column chromatography (dichloromethane:methanol (v:v) = 1:10) to give compound 8D (6.4 g, yield: 87.58%). LCMS (ESI): m / z = 169.1 [M+H] + .

[0255] Step 5: Compound 8D (6.4 g, 38.05 mmol) was added to a flask, along with acetonitrile (100 mL) and copper dichloride (10.23 g, 76.1 mmol). The mixture was then reacted at 85 °C for 1 hour. After cooling, the mixture was directly concentrated, and the residue was purified by column chromatography (dichloromethane:methanol (v:v) = 1:5) to give compound 8E (4.0 g, yield: 63.26%). LCMS (ESI): m / z = 167.1 [M+H] + .

[0256] Step 6: Compound 8E (4 g, 24.07 mmol) was added to a flask, followed by acetonitrile (60 mL), cesium carbonate (15.68 g, 48.14 mmol), and 4-chlorobenzyl chloride (5.81 g, 36.11 mmol). The mixture was then reacted at 85 °C for 2 hours. After cooling, the mixture was filtered directly. The yield filtrate was concentrated, and the residue was purified by column chromatography (dichloromethane:methanol (v:v) = 10:1) to give compound 8F (6.3 g, yield: 90.02%). LCMS (ESI): m / z = 291.1 [M+H] + .

[0257] Step 7: Add 40 mL of anhydrous dichloromethane to a flask, then add 1.2 mL of oxaloyl chloride (14.42 mmol). Under nitrogen protection, add 1.88 mL of anhydrous dimethyl sulfoxide (26.49 mmol) dropwise at -78 °C. After the addition is complete, continue the reaction for 30 minutes. Then, dissolve 3.5 g of compound 8F (12.04 mmol) in 30 mL of dichloromethane and add it dropwise to the above system. After the addition is complete, react at -78 °C for 1 hour. Then, add 13.35 mL of triethylamine (96.31 mmol) dropwise and react for another hour. Add water to the reaction system, extract with dichloromethane, collect the organic phase, wash, and use the resulting dichloromethane solution of compound 8G directly for the next reaction. LCMS (ESI): m / z = 289.1 [M+H] + .

[0258] Step 8: Add the dichloromethane solution of compound 8g from the previous step to the flask, add 20 mL of N,N-dimethylformamide dimethyl acetal, and then concentrate to remove the dichloromethane. Continue reacting the remaining solution in the flask at 110°C for 1 hour. After cooling, concentrate directly to obtain compound 8H (5.2 g crude product), which can be used directly in the next reaction. LCMS (ESI): m / z = 344.3 [M+H] + .

[0259] Step 9: Compound 8H (3.5 g, 12.04 mmol) was added to a flask, along with dioxane (30 mL), S-methylisothiourea sulfate (13.62 g, 72.24 mmol), and potassium carbonate (19.94 g, 144.48 mmol). The mixture was then reacted at 110 °C for 5 hours. After cooling, the mixture was filtered directly. The filtrate was concentrated, and the residue was purified by column chromatography (petroleum ether:ethyl acetate (v:v) = 1:5) to give compound 8I (1.2 g, yield: 26.93%). LCMS (ESI): m / z = 371.1 [M+H] + .

[0260] Step 10: Add 8I (200 mg, 0.54 mmol) to a flask, along with dichloromethane (8 mL) and m-chloroperoxybenzoic acid (131 mg, 0.65 mmol). React at room temperature for 16 hours. Add saturated sodium thiosulfate solution to the reaction mixture, extract with dichloromethane, collect the organic phase, wash and concentrate. Purify the residue by column chromatography (ethyl acetate) to give compound 8J (120 mg, yield: 55.16%). LCMS (ESI): m / z = 403.3 [M+H] + .

[0261] Step 11: Add 8 J (90 mg, 0.22 mmol) to a flask, along with acetonitrile (10 mL), 2,2,2-trifluoroethanol (88 mg, 0.88 mmol), and potassium carbonate (91 mg, 0.66 mmol). React at 50 °C for 1 hour. After cooling, filter directly. Concentrate the filtrate and purify the residue by preparative HPLC to obtain compound 8 (41 mg, yield: 44.08%). 1 H NMR(400MHz, CDCl3)δ9.08(s,1H),7.41(d,2H),7.31(d,2H),6.80(d,1H),5.33(d,2 H),4.86(t,2H),3.05-3.02(m,2H),2.97-2.94(m,2H); LCMS(ESI):m / z=423.0[M+H] + .

[0262] Example 9

[0263]

[0264] Step 1: Compound 9A (10.00 g, 64.86 mmol), potassium carbonate (22.41 g, 162.11 mmol), and 3-chloro-5-chloromethylpyridine (10.51 g, 64.86 mmol) were sequentially added to acetonitrile (50 mL) solvent, and the mixture was heated to 80 °C and reacted for two hours. After the reaction was complete, ethyl acetate and saturated brine were added, and the mixture was separated and washed. The organic phase was dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel column chromatography to give compound 9B (15.20 g, yield: 83.76%). LC-MS (ESI): m / z = 280.20 [M+H] + .

[0265] Step 2: Compound 9B (15.2 g, 54.35 mmol) and lithium hydroxide monohydrate (4.56 g, 108.70 mmol) were added to methanol (50 mL) solvent. Distilled water (20 mL) was added with stirring, and the mixture was heated to 50 °C and reacted for two hours. After the reaction was complete, dichloromethane and distilled water were added, and the mixture was 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 9C (13.6 g, yield: 94.20%). LC-MS (ESI): m / z = 266.20 [M+H] + .

[0266] Step 3: Compound 9C (32.00 g, 105.93 mmol) was added to dichloromethane (300 mL) solvent, followed by N,N-dimethylformamide (0.5 mL). The mixture was reacted dropwise with oxaloyl chloride (26.89 g, 211.86 mmol) under a nitrogen atmosphere on an ice bath for 2 hours at room temperature. After the reaction was complete, the reaction solution was concentrated and dissolved in dichloromethane (100 mL) to obtain a dichloromethane solution of the acyl chloride. Dimethylhydroxylamine hydrochloride (15.50 g, 158.90 mmol) and triethylamine (32.16 g, 317.79 mmol) were added to dichloromethane (200 mL) solvent. The prepared dichloromethane solution of the acyl chloride was then slowly added dropwise to the reaction solution under an ice bath. The reaction was continued at room temperature for 1 hour. After the reaction was complete, distilled water was added, and the mixture was separated and washed. The resulting organic phase was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to give compound 9D (32.00 g, yield: 97.86%). LC-MS (ESI): m / z = 309.0 [M+H] + .

[0267] Step 4: Compound 9D (32.00 g, 103.65 mmol) was added to tetrahydrofuran (200 mL) solvent. Under nitrogen protection, the temperature was lowered to -15 °C, and methyl magnesium chloride (3 mol / L, 38.01 mL) tetrahydrofuran solution was slowly added dropwise. After the addition was complete, the reaction was allowed to proceed for 2 hours. After the reaction was complete, ethyl acetate and saturated ammonium chloride aqueous solution were added, and the mixture was 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 give compound 9E (16.70 g, yield: 61.10%). LC-MS (ESI): m / z = 264.20 [M+H] + .

[0268] Step 5: Compound 9E (16.70 g, 63.57 mmol) was added to dimethyl sulfoxide (100 mL) solvent, followed by 48% hydrobromic acid (16.07 g, 95.36 mmol). The mixture was heated to 110 °C and reacted for 2 hours. After the reaction was complete, ethyl acetate and saturated brine were added, and the mixture was 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 give compound 9F (15 g, yield: 85.28%). LC-MS (ESI): m / z = 275.1 [MH] - .

[0269] Step 6: Compound 9F (5.00 g, 18.07 mmol) was added to toluene (50 mL) solvent, followed by trimethyl orthoformate (2.30 g, 21.68 mmol) and p-toluenesulfonic acid (3.73 mg, 21.68 mmol). The mixture was heated to 110 °C and reacted for 2 hours. After the reaction was complete, ethyl acetate and saturated brine were added, and the mixture was separated and washed. The organic phase was dried over anhydrous sodium sulfate and concentrated to dryness under reduced pressure. The residue was purified by silica gel column chromatography to give compound 9G (5.10 g, yield: 87.44%). LC-MS (ESI): m / z = 323.50 [M+H] + .

[0270] Step 7: Compound 9G (2.00 g, 6.20 mmol) was added to ethanol (20 mL), followed by aminoguanidine hydrochloride (0.75 g, 6.82 mmol) and p-toluenesulfonic acid (0.21 g, 1.24 mmol). The mixture was heated to 80 °C and reacted overnight. After the reaction was complete, dichloromethane and distilled water were added, and the mixture was 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 give compound 9H (1.9 g, yield: 97.42%). LC-MS (ESI): m / z = 379.1 [M+H] + .

[0271] Step 8: Compound 9H (1.9 g, 5.03 mmol) was added to acetic acid (20 mL) solvent, followed by p-toluenesulfonic acid (0.21 g, 1.24 mmol). The mixture was heated to 120 °C and reacted for 24 hours. After the reaction was complete, the reaction solution was concentrated under reduced pressure, and dichloromethane and distilled water were added. The mixture was 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 9J (1.5 g, yield: 95.00%). LC-MS (ESI): m / z = 315.30 [M+H] + .

[0272] Step 9: Compound 9J (1.50 g, 4.78 mmol) and copper bromide (1.60 g, 7.17 mmol) were added to acetonitrile (20 mL) solvent. Under nitrogen protection, the temperature was lowered to -15 °C, and tert-butyl nitrite (0.74 g, 7.17 mmol) was slowly added dropwise. After the addition was complete, the mixture was slowly brought to room temperature and stirred for 30 minutes. After the reaction was complete, ethyl acetate and water were added, and the mixture was washed separately. The organic phase was dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel column chromatography to give compound 9K (520 mg, yield: 28.80%). LC-MS (ESI): m / z = 378.0, 380.0 [M+H] + .

[0273] Step 10: 2,2,2-Trifluoroethanol (78 mg, 0.78 mmol) was added to tetrahydrofuran (10 mL) solvent, followed by sodium hydride (19.92 mg, 0.52 mmol). The mixture was stirred at room temperature for 30 minutes. Compound 9K (100 mg, 0.26 mmol) was added, and the mixture was reacted at room temperature for 2 hours. After the reaction was complete, ethyl acetate and saturated ammonium chloride aqueous solution were added. The mixture was washed separately, and the organic phase was dried over anhydrous sodium sulfate and concentrated to dryness. The residue was purified by reversed-phase silica gel column chromatography to give compound 9 (65 mg, yield: 61.87%). 1 H NMR(400MHz, CDCl3)δ9.22(s,1H),8.37-8.31(m,1H),7.45–7.37(m,2H),7.37–7.28(m, 2H),7.15-7.08(m,1H),5.38(s,2H),5.08-4.98(m,2H); LC-MS(ESI):m / z=398.30[M+H] + .

[0274] Example 10

[0275]

[0276] Step 1: Compound 10A (10 g, 47.96 mmol) and 2,2-difluoroethanol (5.51 g, 67.1 mmol) were dissolved in N,N-dimethylformamide (80 mL). The mixture was then cooled to 0°C, and sodium hydride (2.53 g, 105.5 mmol) was added in portions. After reacting at room temperature for half an hour, the mixture was reacted at 90°C for 3 hours. After the reaction was complete, the reaction solution was diluted with water, extracted three times with ethyl acetate, and the combined organic phases were concentrated. The residue was purified by silica gel column chromatography to give compound 10B (7.7 g, yield: 63%). LC-MS (ESI): m / z = 253.7 [M+H] + .

[0277] Step 2: Compound 10B (7.6 g, 29.9 mmol) was dissolved in pyridine hydrofluoride solution (80 mL), and sodium nitrite (3.10 g, 44.88 mmol) was added in portions at 0°C. The reaction was continued for 15 min, then allowed to return to room temperature for 15 min, followed by heating to 35°C for 3 hours. After completion, the reaction solution was slowly added to a saturated sodium carbonate solution, and the mixture was extracted three times with ethyl acetate. The combined organic phases were concentrated, and the residue was purified by silica gel column chromatography to give compound 10C (6 g, yield: 78%). LC-MS (ESI): m / z = 256.8 [M+H] + .

[0278] Step 3: Compound 10C (7 g, 27.24 mmol), dipentanoyl diboron (8.99 g, 35.41 mmol), potassium acetate (6.68 g, 68.1 mmol), and XPhos Pd G2 (2.14 g, 2.72 mmol) were dissolved in 1,4-dioxane (70 mL), then purged three times with nitrogen, and heated to 100 °C with stirring for 3 hours under a nitrogen atmosphere. The reaction solution was filtered while hot, and the filtrate was directly concentrated to obtain compound 10D (6 g crude product), which was used directly in the next step of the reaction. LC-MS (ESI): m / z = 223.2 [M+H] + .

[0279] Step 4: Compound 10D (6 g, 27.04 mmol), 6-bromo-2,3-dihydropyridazin-3-one (4.73 g, 27.04 mmol), tripotassium phosphate (5.74 g, 27.04 mmol), and [1,3-bis(2,6-diisopropylphenyl)imidazol-2-ylyl](3-chloropyridinyl)palladium(II) dichloride (PEPPSI-IPR catalyst) (1.8 g, 2.7 mmol) were dissolved in 1,4-dioxane (60 mL) and water (20 mL). The mixture was exchanged with nitrogen three times, and then heated to 90 °C and stirred for 3 hours under a nitrogen atmosphere. After the reaction was complete, the mixture was filtered, and the filtrate was extracted three times with ethyl acetate. The combined organic phases were concentrated, and the residue was purified by silica gel column chromatography to give compound 10E (7 g, yield: 95%). LC-MS (ESI): m / z = 273.1 [M+H] + .

[0280] Step 5: Compound 10E (0.25 g, 0.92 mmol), 1-(bromomethyl)-4-chlorobenzene (0.25 g, 1.21 mmol), and potassium carbonate (0.32 g, 2.32 mmol) were dissolved in N,N-dimethylformamide (8 mL), and then stirred at 70 °C for 3 hours. After the reaction was complete, the mixture was diluted with water, extracted three times with ethyl acetate, and the organic phases were combined and concentrated. The residue was sent for preparative HPLC purification to obtain compound 10 (27 mg, yield: 7.4%). 1 H NMR(400MHz,DMSO-d6)δ8.76(d,1H),8.02(d,1H),7.45–7.40(m,4H),7.13(d,1H) ,6.65-6.26(m,1H),5.33(s,2H),4.84-4.69(m,2H); LC-MS(ESI):m / z=397.1[M+H] + .

[0281] Biological testing

[0282] I. Methods for Analyzing Rabbit Skeletal Muscle Myosin Activity

[0283] The purpose of this experiment was to evaluate the ability of the compound to inhibit the hydrolysis of ATP by rabbit skeletal muscle myosin. The assay method is as follows:

[0284] Solution preparation:

[0285] • Prepared F-actin filaments (#AKF99-B, Cytoskeleton) were diluted to 1 mg / mL with reaction buffer (25 mM Tris-HCl, pH 7.5 + 35 mM KCl + 0.1 mM EGTA + 1 mM MgCl2);

[0286] Rabbit myosin (#MY02-A, Cytoskeleton) was diluted to 0.1 mg / mL with resuspension buffer (15 mM Tris-HCl, pH 7.5 + 0.2 M KCl + 1 mM MgCl2);

[0287] • ATP (#BSA04-001, Cytoskeleton) was diluted with 15 mM Tris-HCl (pH = 7.5) to prepare a 3 mM working solution;

[0288] Experimental steps:

[0289] • Add 13 μL of reaction buffer + 12 μL of F-actin to the actin control well;

[0290] • Add 10 μL reaction buffer + 12 μL F-actin + 3 μL myosin II to the positive control well;

[0291] • Add 10 μL of compound working solution + 12 μL of F-actin + 3 μL of myosin II to the compound well;

[0292] • Add 5 μL of ATP working solution to all test wells to start the reaction. Add Pi standard solution to the wells simultaneously according to the instructions (#BK054, Cytoskeleton).

[0293] After gently mixing, incubate at 37°C for 60 minutes.

[0294] • Add 70 μL of CytoPhos Reagent (#BK054, Cytoskeleton) to each well and incubate for 10 min to terminate the reaction.

[0295] Detection:

[0296] The OD value of each well was measured at 650 nm using the endpoint method. A standard curve was fitted based on the Pi concentration and OD value, and the Pi concentration of each test well was calculated. Inhibition (%) = (Pi positive control wells - Pi test wells) / Pi positive control wells * 100%. The Inhibition % - Enzyme concentration curve was fitted using Graphpad Prism 8 software, and the IC50 was calculated. 50 .

[0297] Detection results: The compound's inhibitory activity against ATP hydrolysis by myosin in rabbit skeletal muscle has an IC50 value. 50 Less than 1000 μM, see Table 1 for specific results.

[0298] Table 1. Inhibitory activity of compounds against rabbit skeletal muscle myosin.

[0299] Compound No. Rabbit skeletal muscle myosin (IC 50 , μM)]]> 1 A 2 A 3 A 4 AA 5 A 6 AA 7 A 8 A 9 A 10 A

[0300] AA indicates 0μM <IC 50 ≤2.5μM, where A represents 2.5μM <IC 50 ≤20μM, where B indicates 20μM <IC 50 ≤50μM, where C represents 50μM <IC 50 ≤100μM, where D represents 100μM <IC 50 ≤200μM.

[0301] Conclusion: The compounds of this invention exhibit strong inhibitory activity against ATP hydrolysis by myosin in rabbit skeletal muscle, and the IC50 of some compounds is [not specified]. 50 Less than 20 μM, for example, the IC50 of compound 1 50 The IC50 of compound 4 was 5.44 μM. 50 The IC50 of compound 5 was 1.31 μM. 50 The IC50 of compound 6 was 2.8 μM. 50 It is 2.1 μM.

[0302] II. Methods for Analyzing Cardiac Myosin SII Activity

[0303] The purpose of this experiment was to evaluate the ability of the compound to inhibit the hydrolysis of ATP by cardiac myosin SII. The assay method is as follows:

[0304] Solution preparation:

[0305] • Prepared F-actin filaments (#AKF99-B, Cytoskeleton) were diluted to 1 mg / mL with reaction buffer (15 mM Tris-HCl, pH 7.5 + 10 mM KCl + 0.1 mM EGTA + 2 mM MgCl2);

[0306] Cardiac Muscle Myosin (#MY03-A, Cytoskeleton) was diluted to 1 mg / mL with Resuspension buffer (15 mM Tris-HCl, pH 7.5 + 0.2 MkCl + 1 mM MgCl2).

[0307] • ATP (#BSA04-001, Cytoskeleton) was diluted with 15 mM Tris-HCl (pH = 7.5) to prepare a 3 mM working solution;

[0308] Preparation of working solution for compound: The final concentration of the compound is 100 μM.

[0309] Experimental steps:

[0310] • Add 10 μL of compound working solution + 12 μL of Preformed F-actin filaments + 3 μL of Cardiac Muscle myosin to the compound well;

[0311] • Add 10 μL of reaction buffer containing DMSO, 12 μL of Preformed F-actinfilaments, and 3 μL of Cardiac Musclemyosin to the positive control well;

[0312] • Add 10 μL of DMSO-containing reaction buffer, 12 μL of Preformed F-actinfilaments, and 3 μL of Resuspension buffer to the negative control well;

[0313] • 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.

[0314] After the reaction is complete, take 6 μL of the reaction solution, add 24 μL of reaction buffer to dilute it 5 times, and then use a phosphate kit (#BK054, Cytoskeleton).

[0315] The content of product Pi was detected;

[0316] • Add Pi standard sample to the orifice according to the instructions;

[0317] • Add 70 μL of CytoPhos Reagent (#BK054, Cytoskeleton) to each well and incubate at room temperature for 10 min before detection.

[0318] Detection:

[0319] The OD value of each well was measured at 650 nm. A standard curve was fitted based on the Pi standard concentration and OD value, and the Pi concentration of each test well was calculated. Inhibition (%) = (positive control wells - test wells) / positive control wells * 100%.

[0320] Table 2. Inhibitory activity of compounds against cardiac myosin SII

[0321] Compound No. Myosin SII (inhibition @ 100 μM) 1 2％ 4 23.1％ 5 20.9％ 7 0％

[0322] Conclusion: The compounds of this invention have almost no inhibitory effect on cardiac myosin SII.

[0323] III. Methods for Analyzing the Activity of S1 Fragment of Smooth Muscle Myosin

[0324] The purpose of this experiment was to evaluate the ability of the compound to inhibit the hydrolysis of ATP by the S1 fragment of smooth muscle myosin. The assay method is as follows:

[0325] Solution preparation

[0326] • F-actin polymer: Cardiac Actin powder (#CS-AD99, Cytoskeleton) was dissolved in buffer (5 mM MPipes-KOH solution, pH=7.0, containing 100 μM ATP and 500 μM DTT) to a concentration of 2 mg / mL, and incubated at room temperature for 30 min; then 2 mM MgCl2 and 2 mM MEGTA were added, and the solution was incubated at room temperature for 40 min.

[0327] • Smooth muscle S1 fragment (#CS-MYS05, Cytoskeleton) was diluted to a solution of 0.7 mg / mL with pre-cooled PM12 buffer (12 mM Pipes-KOH solution, pH=7.0, containing 2 mM MgCl2) containing 1 mM DTT.

[0328] • ATP (#BSA04-001, Cytoskeleton) was diluted with 15 mM Tris-HCl (pH = 7.5) to prepare a 5 mM working solution;

[0329] Compound working solution: The final concentration of the compound is 100 μM

[0330] Experimental steps:

[0331] • Add 10 μL of compound working solution + 6 μL of F-actin polymer + 4 μL of Smooth muscle S1fragment solution to the compound well;

[0332] • Add 10 μL of PM12 buffer containing DMSO + 6 μL of F-actin polymer + 4 μL of Smoothmuscle S1 fragment solution to the positive control well;

[0334] • Add 10 μL of PM12 buffer containing DMSO, 6 μL of F-actin polymer, and 4 μL of PM12 buffer to the negative control well;

[0335] • 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 min.

[0336] After the reaction is complete, take 6 μL of the reaction solution, add 24 μL of PM12 buffer, dilute 5 times, and then use a phosphate kit (#BK054, Cytoskeleton).

[0337] The content of product Pi was detected;

[0338] • Add the standard according to the instructions. Add 70 μL of CytoPhos Reagent (#BK054, Cytoskeleton) to all wells and incubate at room temperature for 10 min before testing.

[0339] Detection: The OD value of each well was measured at 650 nm. A standard curve was fitted based on the Pi standard concentration and OD value, and the Pi concentration of each test well was calculated. Inhibition (%) = (Positive control wells - Test wells) / Positive control wells * 100%.

[0340] Conclusion: The compounds of the present invention, such as the compounds in the examples, have almost no inhibitory effect on smooth muscle myosin S1.

[0341] IV. Mouse Pharmacokinetic Test

[0342] 1. Experimental animals: Male C57 mice, 22-25g, 9 mice / compound. Purchased from Beijing Huafukang Biotechnology Co., Ltd.

[0343] 2. Experimental Design: On the day of the experiment, C57 mice were randomly divided into groups according to body weight. They were fasted for 12–14 hours before administration but allowed free access to water. Food was given 4 hours after administration. Administration was performed according to Table 3.

[0344] Table 3 Dosage Information

[0345]

[0346] Note: Intravenous administration solvent: 5% DMA + 5% Solutol + 90% Saline; Gavage administration solvent: 0.5% MC

[0347] (DMA: dimethylacetamide; Solutol: polyethylene glycol-15-hydroxystearate; Saline: physiological saline; MC: methylcellulose)

[0348] Blood samples of 0.06 mL were collected via the orbital cavity before and after isoflurane anesthesia, placed in EDTAK2 centrifuge tubes, and centrifuged at 5000 rpm for 10 min at 4°C to collect plasma. Blood collection time points for both the intravenous and gavage groups were 0, 5, 15, 30 min, 1, 2, 4, 7, 24, and 48 h. All samples were stored at -80°C before analysis and quantitative analysis was performed using LC-MS / MS.

[0349] Table 4. Pharmacokinetic parameters of the tested compounds in mouse plasma.

[0350]

[0351] Compare the structure of compound A:

[0352] Conclusion: The compounds of the present invention, such as the compounds in the examples, have favorable pharmacokinetic characteristics in mice.

[0353] V. Rat Pharmacokinetic Tests

[0354] 1. Experimental animals: Male SD rats, approximately 220g, 6-8 weeks old, 6 rats / compound. Purchased from Chengdu Dashuo Experimental Animal Co., Ltd.

[0355] 2. Experimental Design: On the day of the experiment, SD rats were randomly divided into groups according to their body weight. They were fasted for 12–14 hours before administration but allowed free access to water. Food was given 4 hours after administration. Administration was performed according to Table 5.

[0356] Table 5. Drug Administration Information

[0357]

[0358] Note: Intravenous administration solvent: 5% DMA + 5% Solutol + 90% Saline; Gavage administration solvent: 0.5% MC

[0359] Blood samples of 0.15 mL were collected via the orbital cavity before and after isoflurane anesthesia, placed in EDTAK2 centrifuge tubes, and centrifuged at 5000 rpm for 10 min at 4°C to collect plasma. Blood collection time points for both the intravenous and gavage groups were 0, 5, 15, 30 min, 1, 2, 4, 6, 8, and 24 h. All samples were stored at -80°C before analysis and quantitative analysis was performed using LC-MS / MS.

[0360] Table 5.1 Pharmacokinetic parameters of the tested compounds in rat plasma

[0361]

[0362] Conclusion: The compounds of the present invention, such as the compounds in the examples, have favorable pharmacokinetic characteristics in rats.

[0363] VI. Pharmacokinetic Testing in Beagle Dogs

[0364] 1. Experimental animals: Male beagle dogs, weighing approximately 8-11 kg, 6 dogs per compound, purchased from Beijing Mars Biotechnology Co., Ltd.

[0365] 2. Experimental Method: On the day of the experiment, beagles were randomly grouped according to their body weight. They were fasted for 12-14 hours prior to administration but allowed free access to water. Food was given 4 hours after administration. Administration was performed according to Table 6.

[0366] Table 6. Drug Administration Information

[0367]

[0368] Note: Intravenous administration solvent: 5% DMA + 5% Solutol + 90% Saline; Gavage administration solvent: 0.5% MC

[0369] Blood samples of 1 mL were collected via jugular or limb veins before and after drug administration and placed in EDTAK2 centrifuge tubes. The plasma was collected by centrifugation at 5000 rpm and 4°C for 10 min. All samples were stored at -80°C before analysis and quantitative analysis was performed using LC-MS / MS.

[0370] Conclusion: The compounds of the present invention, such as the compounds in the examples, have favorable pharmacokinetic characteristics in beagle dogs.

[0371] VII. Pharmacokinetic Tests in Monkeys

[0372] 1. Experimental animals: Male cynomolgus monkeys, 3-5 kg, 3-6 years old, 4 per compound. Purchased from Suzhou Xishan Biotechnology Co., Ltd.

[0373] 2. Experimental Methods: On the day of the experiment, monkeys were randomly divided into groups according to their body weight. They were fasted for 14–18 hours prior to drug administration but allowed free access to water. Food was given 4 hours after drug administration. Drug administration was performed according to Table 7.

[0374] Table 7 Dosage Information

[0375]

[0376] Note: Intravenous administration solvent: 5% DMA + 5% Solutol + 90% Saline; Gavage administration solvent: 0.5% MC

[0377] Blood samples of 1.0 mL were collected from venous sites in the extremities before and after drug administration and placed in EDTAK2 centrifuge tubes. The plasma was collected by centrifugation at 5000 rpm and 4°C for 10 min. All samples were stored at -80°C before analysis and quantitative analysis was performed using LC-MS / MS.

[0378] Conclusion: The compounds of the present invention, such as the compounds in the examples, have favorable pharmacokinetic characteristics in monkeys.

[0379] VIII. hERG Potassium Ion Channel Function Test

[0380] 1. Experimental platform: Electrophysiological manual patch-clamp system

[0381] 2. Cell line: Chinese hamster ovary (CHO) cell line stably expressing hERG potassium ion channels

[0382] 3. Experimental Methods: CHO (Chinese Hamster Ovary) cells stably expressing hERG potassium channels were used to record hERG potassium channel currents at room temperature using whole-cell patch-clamp technique. Glass microelectrodes were fabricated from glass electrode blanks (BF150-86-10, Sutter) using a stretching device. The tip resistance after perfusion with electrode fluid was approximately 2-5 MΩ. The glass microelectrodes were inserted into the amplifier probe to connect to the patch-clamp amplifier. Clamp voltage and data recording were controlled and recorded using pClamp 10 software via computer, with a sampling frequency of 10 kHz and a filtering frequency of 2 kHz. After obtaining whole-cell recordings, the cells were clamped at -80 mV to induce hERG potassium current (Ig). hERG The step voltage was applied from -80 mV to +20 mV for 2 seconds, then repolarized to -50 mV for 1 second before returning to -80 mV. This voltage stimulation was applied every 10 seconds, and the drug administration process began after the hERG potassium current stabilized (at least 1 minute). Each test concentration of the compound was administered for at least 1 minute, and at least 2 cells (n≥2) were tested for each concentration.

[0383] 4. Data Processing: Data analysis and processing were performed using pClamp 10, GraphPad Prism 5, and Excel software. The degree of inhibition of hERG potassium current (the peak hERG tail current induced at -50mV) by different compound concentrations was calculated using the following formula:

[0384] Inhibition%=[1–(I / Io)]×100%

[0385] Where Inhibition% represents the percentage of inhibition of hERG potassium current by the compound, and I and Io represent the amplitude of hERG potassium current before and after drug administration, respectively.

[0386] Compound IC 50 The following equations were fitted and calculated using GraphPad Prism 5 software:

[0387] Y=Bottom+(Top-Bottom) / (1+10^((LogIC50-X)*HillSlope))

[0388] Where X is the Log value of the detected concentration of the test sample, Y is the inhibition percentage at the corresponding concentration, and Bottom and Top are the minimum and maximum inhibition percentages, respectively.

[0389] Table 8 shows the IC50 values ​​of the compounds on the inhibitory effect of hERG potassium channel current. 50

[0390] Test Compound No. IC50(μM) 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 1 >10 2 ＞10 5 >10

[0391] Conclusion: The compounds of the present invention, such as the compounds in the examples, do not inhibit the hERG potassium channel current.

[0392] IX. CYP450 Enzyme Inhibition Test

[0393] The aim of this study was to evaluate the effects of test substances on the activities of five isoenzymes (CYP1A2, CYP2C9, CYP2C19, CYP2D6, and CYP3A4) of human liver microsomal cytochrome P450 (CYP) using an in vitro assay system. Specific probe substrates for CYP450 isoenzymes were co-incubated with human liver microsomes and different concentrations of the test substances. Reduced nicotinamide adenine dinucleotide phosphate (NADPH) was added to initiate the reaction. After the reaction, the metabolites produced by the specific substrates were quantitatively detected by liquid chromatography-tandem mass spectrometry (LC-MS / MS) after sample processing, and the changes in CYP enzyme activity were measured. The IC50 values ​​were calculated. 50 The value is used to evaluate the inhibitory potential of the test substance against each CYP enzyme subtype.

[0394] Conclusion: The compounds of the present invention, such as the compounds in the examples, do not inhibit the five isoenzymes of human liver microsomal cytochrome P450 (CYP).

[0395] 10. Caco-2 Permeability Test

[0396] The experiment used monolayers of Caco-2 cells, incubated in triple parallel in 96-well Transwell plates. A transport buffer solution (HBSS, 10 mM HEPES, pH 7.4±0.05) containing either the compound of the present invention (2 μM) or the control compounds digoxin (10 μM), naldolol (2 μM), and metoprolol (2 μM) was added to the dosing well on the apical or basal side. A transport buffer solution containing DMSO was added to the corresponding receiving well. After incubation at 37±1 °C for 2 hours, the cell plate was removed, and appropriate amounts of sample were transferred from both the apical and basal sides to new 96-well plates. Acetonitrile containing an internal standard was then added to precipitate the protein. The samples were analyzed using LC MS / MS to determine the concentrations of the compound of the present invention and the control compounds. The concentration data were used to calculate the apparent permeability coefficients for transport from the apical to the basal side of the monolayer cells, and from the basal side to the apical side, thereby calculating the efflux rate. Leakage of fluorescein was used to evaluate the integrity of the monolayer cells after 2 hours of incubation.

[0397] Conclusion: The compounds of the present invention, such as the compounds in the examples, exhibited good permeability in the Caco-2 permeability test.

[0398] XI. Liver microsomal stability test

[0399] This experiment used liver microsomes from five genera—human, canine, rat, and mouse—as in vitro models to evaluate the metabolic stability of the test substance.

[0400] At 37°C, 1 μM of the test substance was co-incubated with microsomal protein and coenzyme NADPH. The reaction was terminated by adding ice-cold acetonitrile containing an internal standard after certain time intervals (5, 10, 20, 30, 60 min). The concentration of the test substance in the sample was detected by LC-MS / MS. T was calculated using the ln value of the drug residue in the incubation system and the incubation time. 1 / 2 Furthermore, the intrinsic clearance rate (CL) of liver microsomes was calculated. int(mic) and hepatic intrinsic clearance

[0401] CL int(Liver) .

[0402] Conclusion: The compounds of the present invention, such as the compounds in the examples, exhibit good metabolic stability in liver microsomal stability tests.

[0403] 12. PK / PD Detection in Dmd(Mdx) Gene Mutant Mouse Model

[0404] Experimental Objective: B10-Dmd-KO (Mdx) mice are mice with loss of function in DMD caused by a frameshift mutation in the Dmd gene. Phenotypic analysis showed that B10-Dmd-KO mice possess a series of characteristics of DMD patients. This study investigated the effects of compound administration on plasma CK and TNNI2 levels in Mdx mice.

[0405] Laboratory animals: B10-Dmd-KO(C57BL / 10ScSnJGpt-Dmdem3Cd4 / Gpt), male, 5-7 weeks old, weighing 28-30g, supplier: Jiangsu Jicui Yaokang Biotechnology Co., Ltd.

[0406] Experimental procedure:

[0407] 1) Running training (D-5): Mdx mice were trained in batches with three time periods: initial speed of 4 m / min, maintained for 2 min, followed by 8 m / min, maintained for 8 min, and finally 12 m / min, maintained for 30 min, for a total of 40 min.

[0408] 2) Grip strength test (D-2): Three tests per round, for a total of three rounds;

[0409] 3) Grouping (D0): After anesthetizing animals with isoflurane (concentration: 5%, gas flow rate: 1L / min), blood (approximately 150μL) was collected via capillary tube through the orbital sinus into a 0.6mL centrifuge tube containing EDTA anticoagulant. Plasma was separated and CK values ​​were measured, which were used as baseline values ​​for grouping. 4) Administration, gripping, and blood collection (D1): The compound was dissolved in solvents of 5% DMA + 5% HS-15 + 90% Saline and administered orally to mice. Then, gripping strength testing was started (three tests per round, for three rounds). One hour after the test, the animals were anesthetized with isoflurane (concentration: 5%, gas flow rate: 1L / min), and blood (approximately 300μL) was collected via capillary tube through the orbital sinus into a 0.6mL centrifuge tube containing EDTA anticoagulant (ensuring blood collection was 6 hours after administration).

[0410] 5) Running, blood collection, and sample collection (D2): 22h and 20min after drug administration, mice were run for 40min (running parameters as above). One hour after the run, mice were anesthetized by inhalation of isoflurane (concentration: 5%, gas flow rate: 1L / min). Blood (≥300μL) was collected through the orbital sinus using a capillary tube into a 0.6mL centrifuge tube containing EDTA anticoagulant. Subsequently, the animals were euthanized, and the tibialis anterior muscle (left leg), soleus muscle (both legs), and myocardium were collected, weighed, and recorded. The samples were then placed in homogenization tubes, flash-frozen in liquid nitrogen, and stored at -80℃.

[0411] Sample processing: EDTA-anticoagulated whole blood was centrifuged at 3000 rpm and 4°C for 10 min, and serum was collected and aliquoted. The separated serum was analyzed for CK and TNNI2 values ​​using a Roche C311 fully automated biochemical analyzer (Mouse TNNI2 ElisaKit, ABBEXA, E2207370Y). Serum concentrations of the compounds, as well as drug concentrations in the tibialis anterior muscle, soleus muscle, and myocardium, were determined by LC / MS.

[0412] Conclusion: The compounds of the present invention, such as those in the examples, have a good inhibitory effect on plasma CK levels in a Dmd(Mdx) gene mutant mouse model.

Claims

1. A compound of formula (I), a stereoisomer, or a pharmaceutically acceptable salt thereof, wherein, A is selected from 4-10 membered heterocycloalkyl, 5-10 membered heteroaryl, 6-10 membered aryl; B is selected from 5-6 membered heteroaryl, 8-10 membered heteroaryl, 5-6 membered heterocycloalkyl; m is selected from 0, 1, 2; p is selected from 0, 1, 2; n is selected from 0, 1, 2.

2. The compound of formula (I) according to claim 1, a stereoisomer, or a pharmaceutically acceptable salt thereof, having the structure of formula (I-1), (I-2), (I-3), (I-4), (I-5), (I-6), (I-7), (I-8), (I-9), (I-10), (I-11), (I-12), (I-13), (I-14), (I-15), (I-16). represents a single or double bond; 3. The compound of formula (I) according to claim 1, a stereoisomer, or a pharmaceutically acceptable salt thereof, wherein, m is selected from 0, 1; p is selected from 0, 1; n is selected from 0, 1; L1 is selected from bond, ethynyl; * indicates the linking position of B ring with L2, * indicates the linking position of B ring with C when L2 is bond, ** indicates the linking position of B ring with L1, ** indicates the linking position of B ring with A ring when L1 is bond.

5. The compound according to claim 1, a stereoisomer, or a pharmaceutically acceptable salt thereof, having the structure of formula (II), (III).

6. The compound according to claim 1, a stereoisomer, or a pharmaceutically acceptable salt thereof, selected from the following structures.

7. A pharmaceutical composition or a pharmaceutical preparation comprising a compound according to any one of claims 1-6, or a stereoisomer, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier and / or excipient.

8. The pharmaceutical composition or the pharmaceutical preparation according to claim 7, comprising 1-1500 mg of a compound according to any one of claims 1-6, or a stereoisomer, or a pharmaceutically acceptable salt thereof, and a carrier and / or excipient. C is selected from C 3-8 alkyl, phenyl, 5-6 membered heteroaryl, C 3-12 carbocyclic, 4-15 membered heterocyclyl; each R 1 is independently selected from the group consisting of H, =0, C 2 1-6 alkyl, haloC 1-4 1-6 alkyl, -O-haloC 1-4 1-6 alkyl, -NH-haloC 1-4 1-6 alkyl, -OC 1-4 1-6 alkyl, C 1-4 1-6 alkyl, C 1-4 1-6 alkyl, halogen, cyano, nitro, OH, C 1-4 1-6 alkyl, C 2-6 1-6 alkyl, -NH-C 1-4 1-6 alkyl, -N(C 1-4 1-6 alkyl)2, -NH-C 3-10 1-6 alkyl, -NHC(O)C 1-4 1-6 alkyl, C 3-10 1-6 alkyl, -O-C 3-10 1-6 alkyl, -O-(4-10 membered heterocycloalkyl), -O-C 1-4 1-6 alkyl, -O-(4-10 membered heterocycloalkyl), -O-C 3-10 1-6 alkyl, amino, or 4-10 membered heterocycloalkyl, said alkyl, alkoxy, cycloalkyl, heterocycloalkyl groups are optionally further substituted with 1-3 groups selected from halogen, D, CN, OH, C 1-4 1-6 alkyl, haloC 1-4 1-6 alkyl, deuterated C 1-4 1-6 alkyl, C 1-4 1-6 alkyl, haloC 1-4 1-6 alkyl, deuterated C 1-4 1-6 alkyl, C 3-10 1-6 alkyl, 4-6 membered heterocycloalkyl, phenyl and NH2; or R 1 and R 2 , or two R 2 adjacent together with the atom linking them form a 5-6 membered carbocyclic ring, a 5-6 membered heterocyclic ring, which is optionally further substituted with 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, C 3-10 cycloalkyl, -O-C 3-10 cycloalkyl, 4-6 membered heterocycloalkyl, phenyl and NH2; each R is independently selected from C 4 each R is independently selected from C 3-10 cycloalkyl, C 2-6 alkenyl, C 2-6 alkynyl, halogen, =0, amino, cyano, -NH-C 1-4 alkyl, -N(C 1-4 alkyl)2, C 1-4 alkyl, C 1-4 alkoxy, halogenated C 1-4 alkyl, 4-10 membered heterocycloalkyl, 5-10 membered heteroaryl, 6-10 membered aryl; R 7 H, halogen, SF5, OH, CN, amino, C 1-4 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-4 alkoxy, halogenated C 1-4 alkyl, =0, =CH2, =CF2, =C(CH3)2, -NH-C 1-4 alkyl, -N(C 1-4 alkyl)2, -S(O)R c , -S(O)2R c , -C(O)R c , -C(O)OR c , -C(O)N(R c )2, -OC(O)R c , -C 0-4 alkyl-(4-10 membered heterocycloalkyl), or -C 0-4 alkyl-(5-10 membered heteroaryl), -P(O)(R c )2, -Si(R c )3, -SF5, -S(O)(=NH)R c , said alkyl, alkenyl, alkynyl, alkoxy, heterocycloalkyl, heteroaryl groups are optionally further substituted with 1-3 groups selected from halogen, 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; each R 6 is independently selected from the group consisting of H, halogen, SF5, OH, CN, amino, C 1-4 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-4 alkoxy, haloC 1-4 alkyl, -NH-C 1-4 alkyl, -N(C 1-4 alkyl)2, -S(O)R c , -S(O)2R c , -C(O)R c , -C(O)OR c , -C(O)N(R c )2, -OC(O)R c , said alkyl, alkenyl, alkynyl, alkoxy groups are optionally further substituted with 1 to 3 groups selected from halogen, deuterium, CN, OH, C 1-4 alkyl, haloC 1-4 alkyl, deuterated C 1-4 alkyl, C 1-4 alkoxy, haloC 1-4 alkoxy, deuterated C 1-4 alkoxy and NH2; each R is independently selected from H, OH, C c each R is independently selected from H, OH, C 1-4 alkyl, C 3-7 cyclopropyl, haloC 1-4 alkyl, deuterated C 1-4 alkyl, C 1-4 alkoxy, haloC 1-4 alkoxy, deuterated C 1-4 alkoxy, NH2; L1is selected from the group consisting of a bond, C 2-6 alkenyl, C 2-6 alkynyl, O, NH, S, -CD2-, -CHD-, -CR L1 R L2 -, C 1-4 alkyl, -C(O)-, S(O), S(O)2, said alkyl being optionally further substituted by R L1 ; L2is selected from the group consisting of a bond, O, NH, S, -CH2-, -CD2-, -CHD-, -CR L1 R L2 -, -CDR L2 -, -CH2C(R L1 )=, -CH2CH=CH-CH=, C 2-4 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, -C(O)-, S(O), S(O)2, said alkyl being optionally further substituted with 1-3 R L1 ; R L1 , R L2 are each independently selected from the group consisting of halogen, OH, CN, amino, C 1-4 alkyl, C 1-4 alkoxy, C 3-6 cycloalkyl, 4-6 membered heterocycloalkyl; R 2 , R 4 , L1and the atom to which they are attached form a 5-7 membered carbocyclic ring, 5-7 membered heterocyclic ring, which is optionally further substituted with 1-3 groups selected from R A3 ; R 4 , R 6 , L2and the atom to which they are attached form a 5-7 membered carbocyclic ring, 5-7 membered heterocyclic ring, which is optionally further substituted with 1-3 groups selected from R A3 ; As an option, two R 6 Substituents, together with the atoms they are attached to, form 4-6 membered carbon rings, 4-6 membered heterocycles, or R 6 R 7 Together with the attached atoms, they form 4-12 membered carbon rings and 4-12 membered heterocycles, wherein the carbon rings and heterocycles are optionally further composed of 1-3 atoms selected from R A3 Substitution of groups; each R is independently selected from =0, halo, deuterium, CN, OH, C A3 each independently selected from =0, halo, deuterium, CN, OH, C 1-4 alkyl, haloC 1-4 alkyl, haloC 1-4 alkyl, C 1-4 alkoxy, haloC 1-4 alkoxy, haloC 1-4 alkoxy, NH2.

9. Use of a compound according to any one of claims 1-6, or a stereoisomer, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition or a pharmaceutical preparation according to claim 7 or 8, for the manufacture of a medicament for the treatment / prevention of a Myosin II mediated disease. each X is independently selected from CR 6 or N.

10. The use according to claim 9, wherein the Myosin II mediated disease is selected from muscular dystrophy. ​ ​ ​ A is selected from R 1 selected from H, =0, C 1-4 alkyl, C 1-4 alkoxy, haloC 1-4 alkyl, -O-haloC 1-4 alkyl, C 3-6 cycloalkyl, -O-C 3-6 cycloalkyl, 4-7 membered heterocycloalkyl, -O-(4-7 membered heterocycloalkyl), -O-C 1-4 alkyl-C 3-6 cycloalkyl, -O-C 1-4 alkyl-(4-7 membered heterocycloalkyl), said alkyl, cycloalkyl, heterocycloalkyl groups being optionally further substituted with 1-3 groups selected from halogen, D, CN, OH, C 1-2 alkyl, haloC 1-2 alkyl, deuteratedC 1-2 alkyl, C 1-2 alkoxy, haloC 1-2 alkoxy, deuteratedC 1-2 alkoxy, C 3-5 cycloalkyl, 4-6 membered heterocycloalkyl, phenyl and NH2groups; each R is independently selected from =0, C 2 each R is independently selected from =0, C 2-4 alkenyl, C 2-4 alkynyl, halo, cyano, nitro, amino, -NH-C 1-4 alkyl, -N(C 1-4 alkyl)2, OH, C 1-4 alkyl, haloC 1-4 alkyl, deuterated C 1-4 alkyl, C 3-6 cycloalkyl, 4-7 membered heterocycloalkyl; B ring is selected from pyridazinone, pyridinone, pyrazinone, pyrimidinone, 5-membered heteroaryl, 8-10-membered bicyclic heteroaryl; each R is independently selected from the group consisting of deuterium, =0, halo, cyano, C 4 alkyl, haloC 1-4 alkyl, haloC 1-4 alkyl, haloC 1-4 alkyl, haloC 1-4 alkyl, haloC 1-4 alkyl, haloC 1-4 alkyl, haloC C is selected from C 5-7 alkyl, each R is independently selected from deuterium, halogen, OH, CN, C 6 each independently selected from deuterium, halogen, OH, CN, C 1-4 alkyl, haloC 1-4 alkyl, haloC 1-4 alkyl, C 1-4 alkoxy, haloC 1-4 alkoxy, haloC 1-4 alkoxy, SF5; R 7 selected from H, halogen, 5-6 membered heteroaryl, C 1-4 alkyl, C 1-4 alkoxy, C 2-4 alkenyl, C 2-4 alkynyl, halogenated C 1-4 alkyl, -C(O)C 1-4 alkyl, =0, =CH2, =CF2, =C(CH3)2, said alkyl, alkenyl, alkynyl, alkoxy, heteroaryl groups are optionally further substituted with 1-3 groups selected from halogen, 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; ​ L2is selected from the group consisting of a bond, -CH2-, -CH2CH2-, -C(O)-, -CHF-, -CF2-, C(CH3)2-, ethynyl, propynyl; * indicates the point of attachment of L2to C.

4. The compound of formula (I) according to claim 1 or 2, a stereoisomer, a pharmaceutically acceptable salt thereof, wherein is selected from wherein represents the site of attachment to L1, which is a bond when, represents the site of attachment to the B ring. selected from ​ selected from or selected from ​ wherein selected from R 7 selected from H, halogen, OH, NH2, CN, =0, =CH2, =CF2, =C(CH3)2, methyl, ethyl, trifluoromethyl, cyclopropyl; R 6 selected from halogen; R 1 selected from R 2 selected from H, halogen; R 4 selected from H; or R 2 , R 4 and the atom to which they are attached form a 5-7 membered carbocyclic ring; ​ selected from ​ ​ ​ ​ ​

Images