Benzimidazole derivatives, preparation methods and medical uses thereof

Abstract

A benzimidazole derivative, its preparation method, and medical use. Specifically, a benzimidazole derivative represented by general formula (I), its preparation method, a pharmaceutical composition containing the derivative, and its use as a therapeutic agent, particularly for treating diseases related to P2X3 activity. #imgabs0#

Description

Technical Field

[0001] The present disclosure relates to the field of medicine, and in particular to a new class of benzimidazole derivatives, a preparation method thereof, and applications in medicine. Background Art

[0002] P2X receptors are a family of cation-permeable, ligand-gated ion channels that open in response to the binding of extracellular adenosine 5'-triphosphate (ATP). They belong to a larger family of receptors known as purinergic receptors. P2X receptors are found in a variety of organisms, including humans, mice, rats, rabbits, chickens, zebrafish, bullfrogs, flukes, and amoebas. Seven separate genes encoding P2X subunits have been identified and designated P2X1 to P2X7. Different subunits exhibit varying sensitivities to purinergic agonists and antagonists.

[0003] The P2X3 receptor has four ATP-binding sites on a single subunit and is composed of two transmembrane domains, an intracellular N-terminus and a conserved sequence located within an extracellular loop. High levels of P2X3 receptor expression have been found in small and medium-diameter neurons specifically involved in nociceptive information. P2X3 receptors also participate in the transmission of some non-noxious sensations. It has been demonstrated that the P2X3 receptor is involved in bladder sensory function and is a key receptor-mediated bladder sensory signaling. It is expressed in the bladder mucosa, which is rich in sensory nerve fibers. P2X3 is also expressed in sensory nerve fibers of the pharyngeal mucosa, where it is involved in the transmission and formation of taste.

[0004] When the body is injured or nerves are damaged, a large amount of ATP is released, activating the presynaptic P2X3 receptors, causing a large amount of Ca2 + Influx of calcium and increased intracellular calcium concentration activate protein kinase A (PKA) and protein kinase C (PKC), leading to phosphorylation of PKA and PKC. This in turn promotes glutamate release, further activating NMDA receptors, leading to excitatory postsynaptic currents and central nervous system sensitization. Numerous studies have shown that upregulation of P2X3 receptor expression can lead to hyperalgesia and participate in pain signaling.

[0005] MK-7264 is a P2X3 receptor antagonist with IC values ​​of 1. 50 The values ​​are ~30nM and 100-250nM respectively. Its use in treating patients with chronic cough has now entered clinical phase III. Summary of the Invention

[0006] The present disclosure provides a compound of formula (I), or a pharmaceutically acceptable salt thereof, or an isomer thereof,

[0007]

[0008] wherein R1 is selected from hydrogen, deuterium, halogen, C1-C6 hydroxyalkyl, C1-C6 alkyl optionally substituted by halogen or deuterium, and C1-C6 alkoxy optionally substituted by halogen or deuterium;

[0009] R2 is each independently selected from hydrogen, deuterium, halogen, C1-C6 hydroxyalkyl, C1-C6 alkyl optionally substituted by halogen or deuterium, and C1-C6 alkoxy optionally substituted by halogen or deuterium;

[0010] R3 and R4 are each independently selected from hydrogen, halogen and C1-C4 alkyl optionally substituted by halogen, or R3 and R4 together with the carbon atom to which they are commonly attached form a C3-C6 cycloalkylene group optionally substituted by halogen, or R3 and R4 on adjacent carbon atoms together form a C3-C8 cycloalkane optionally substituted by halogen;

[0011] R5 is selected from C1-C6 alkyl optionally substituted by halogen or cyano, C3-C6 cycloalkyl optionally substituted by halogen or cyano, heterocyclyl optionally substituted by halogen or cyano, C1-C6 alkoxy optionally substituted by halogen or cyano, and amino optionally substituted by alkyl;

[0012] R6 are each independently selected from hydrogen, deuterium, halogen, cyano, cyclopropyl and C1-C6 alkyl optionally substituted by halogen or deuterium;

[0013] R7 and R8 are each independently selected from:

[0014] a) hydrogen, deuterium, halogen, cyano, amino, hydroxy, C1-C6 alkyl optionally substituted by halogen, sulfone, sulfoxide, sulfonamide, sulfenamide, C 1-3 and a C1-C6 alkoxy group optionally substituted by halogen;

[0015] b) p is selected from 0, 1 and 2, R9 and R 10 are independently selected from hydrogen, C1-C6 alkyl, C1-C6 alkoxy and C3-C8 cycloalkyl, or R9 and R 10 Together with the nitrogen atom to which they are commonly attached, they form a 4-6 membered heterocyclic group, which is optionally substituted with one or more substituents selected from halogen, hydroxy, cyano, C1-C6 haloalkyl and C1-C6 alkyl; R' is selected from hydrogen, C1-C6 alkyl, C3-C8 cycloalkyl, aryl and heteroaryl; and When p is 0, R9 and R 10 The combination of is not hydrogen and methyl;

[0016] c)

[0017] d) heterocyclyl and heteroaryl, each of which is optionally substituted with one or more substituents selected from oxo, halogen, hydroxy, carbonyl, C1-C6 alkyl and cyano, wherein the C1-C6 alkyl is optionally substituted with one or more halogen; and

[0018] e) Among them, R 11 is selected from C1-C6 alkyl, C1-C6 alkoxy, aryl, heteroaryl, C3-C8 cycloalkyl, heterocyclyl, C1-C6 cyanoalkyl, C3-C8 cycloalkyloxy and amino optionally substituted with C1-C6 alkyl; or

[0019] R7 and R8 together with the atoms to which they are attached form an optionally substituted aromatic or non-aromatic heterocyclic ring;

[0020] X is selected from oxygen atoms, -NH- and methylene, wherein the methylene is optionally substituted with one or more substituents selected from halogen, C3-C8 cycloalkyl, C3-C6 cycloalkylene and C1-C6 alkyl;

[0021] m is an integer from 1 to 3; and

[0022] n is an integer from 1 to 4.

[0023] In some embodiments, R7 and R8 are each independently selected from:

[0024] a) hydrogen, deuterium, halogen, cyano, amino, sulfone, sulfonamide, sulfenamide and C1-C3 alkyl substituted by one or more halogens;

[0025] b) p is selected from 0, 1 and 2, R9 and R 10 are independently selected from hydrogen, C1-C3 alkyl, C1-C3 alkoxy and C3-C6 cycloalkyl, or R9 and R 10 Together with the nitrogen atom to which they are commonly attached, they form a 4-6 membered heterocyclic group, which is optionally substituted with one or more substituents selected from halogen, hydroxyl and C1-C3 alkyl; R' is selected from hydrogen, C1-C3 alkyl, C3-C6 cycloalkyl, aryl and heteroaryl; and When p is 0, R9 and R 10 The combination of is not hydrogen and methyl;

[0026] c)

[0027] d) 4-6 membered heterocyclic or heteroaryl, said heterocyclic or heteroaryl being optionally substituted by one or more substituents selected from oxo, halogen, hydroxy, carbonyl, C1-C3 alkyl and cyano, said C1-C3 alkyl being optionally substituted by one or more halogen; and

[0028] e) Among them, R 11 Selected from C1-C3 alkyl, C1-C3 alkoxy, 5-6 membered aryl or heteroaryl, 3-8 membered cycloalkyl, 3-8 membered heterocyclyl, C1-C3 cyanoalkyl, C3-C6 cycloalkyloxy and amino optionally substituted by C1-C3 alkyl.

[0029] In some embodiments, R7 is a 4-6 membered heterocyclyl or heteroaryl, wherein the heterocyclyl or heteroaryl is optionally substituted with one or more substituents selected from oxo, halogen, hydroxy, carbonyl, C1-C3 alkyl and cyano, wherein the C1-C3 alkyl is optionally substituted with one or more halogen; and

[0030] R8 is selected from hydrogen, deuterium, halogen, cyano and C1-C3 alkyl substituted by one or more halogens.

[0031] In some embodiments, R7 is a 4-6 membered heterocyclyl comprising -NH-C(=O)- or -NH-S(=O)2-, wherein the heterocyclyl is optionally substituted with one or more substituents selected from oxo, halogen, hydroxy, carbonyl, C1-C3 alkyl and cyano, wherein the C1-C3 alkyl is optionally substituted with one or more halogen; and

[0032] R8 is selected from hydrogen, deuterium, halogen, cyano and C1-C3 alkyl substituted by one or more halogens.

[0033] In some embodiments, R7 is a 5-membered heterocyclyl comprising -NH-C(=O)-, which is optionally substituted with one or more substituents selected from oxo, halogen, hydroxy, carbonyl, C1-C3 alkyl, and cyano, wherein the C1-C3 alkyl is optionally substituted with one or more halogen; and

[0034] R8 is selected from hydrogen, deuterium, halogen, cyano and C1-C3 alkyl substituted by one or more halogens.

[0035] In some embodiments, in the compound of formula (I),

[0036] R1 is selected from hydrogen, deuterium, halogen, C1-C6 hydroxyalkyl, C1-C6 alkyl optionally substituted by halogen or deuterium, and C1-C6 alkoxy optionally substituted by halogen or deuterium;

[0037] R2 is each independently selected from hydrogen, deuterium, halogen, C1-C6 hydroxyalkyl, C1-C6 alkyl optionally substituted by halogen or deuterium, and C1-C6 alkoxy optionally substituted by halogen or deuterium;

[0038] R3 and R4 are each independently selected from hydrogen, halogen and C1-C4 alkyl optionally substituted by halogen, or R3 and R4 together with the carbon atom to which they are commonly attached form a C3-C6 cycloalkylene group optionally substituted by halogen, or R3 and R4 on adjacent carbon atoms together form a C3-C8 cycloalkane optionally substituted by halogen;

[0039] R5 is selected from C1-C6 alkyl optionally substituted by halogen or cyano, C3-C6 cycloalkyl optionally substituted by halogen or cyano, heterocyclyl optionally substituted by halogen or cyano, C1-C6 alkoxy optionally substituted by halogen or cyano, and amino optionally substituted by alkyl;

[0040] R6 are each independently selected from hydrogen, deuterium, halogen, cyano, cyclopropyl and C1-C6 alkyl optionally substituted by halogen or deuterium;

[0041] R7 and R8 together with the atoms to which they are attached form a 3-12 membered aromatic or non-aromatic heterocyclic ring, said heterocyclic ring being monocyclic or bicyclic, said heterocyclic ring being optionally substituted with one or more substituents selected from C1-C6 alkylamide, halogen, oxo, C1-C6 alkyl optionally substituted with halogen, and C1-C6 alkoxy;

[0042] X is selected from oxygen atoms, -NH- and methylene, wherein the methylene is optionally substituted with one or more substituents selected from halogen, C3-C8 cycloalkyl, C3-C6 cycloalkylene and C1-C6 alkyl;

[0043] m is an integer from 1 to 3; and

[0044] n is an integer from 1 to 4.

[0045] In some embodiments, R7 and R8 together with the atoms to which they are attached form a 3-12 membered non-aromatic heterocyclic ring, which is monocyclic or bicyclic, and which contains -NH-C(=O)- or -NH-S(=O)2-, which is optionally substituted with one or more substituents selected from C1-C6 alkylamide, halogen, oxo, C1-C6 alkyl optionally substituted with halogen, and C1-C6 alkoxy.

[0046] In some embodiments, R7 and R8 together with the atoms to which they are attached form a 4-8 membered non-aromatic heterocyclic ring, which is monocyclic or bicyclic, and which contains -NH-C(=O)- or -NH-S(=O)2-, which is optionally substituted with one or more substituents selected from C1-C3 alkylamide, halogen, oxo, C1-C3 alkyl optionally substituted with halogen, and C1-C3 alkoxy.

[0047] In some embodiments, in the compound of formula (I),

[0048] R1 is selected from hydrogen, deuterium, halogen, C1-C6 hydroxyalkyl, C1-C6 alkyl optionally substituted by halogen or deuterium, and C1-C6 alkoxy optionally substituted by halogen or deuterium;

[0049] R2 is each independently selected from hydrogen, deuterium, halogen, C1-C6 hydroxyalkyl, C1-C6 alkyl optionally substituted by halogen or deuterium, and C1-C6 alkoxy optionally substituted by halogen or deuterium;

[0050] R3 and R4 are each independently selected from hydrogen, halogen and C1-C4 alkyl optionally substituted by halogen, or R3 and R4 together with the carbon atom to which they are commonly attached form a C3-C6 cycloalkylene group optionally substituted by halogen, or R3 and R4 on adjacent carbon atoms together form a C3-C8 cycloalkane optionally substituted by halogen;

[0051] R5 is selected from C1-C6 alkyl optionally substituted by halogen or cyano, C3-C6 cycloalkyl optionally substituted by halogen or cyano, heterocyclyl optionally substituted by halogen or cyano, C1-C6 alkoxy optionally substituted by halogen or cyano, and amino optionally substituted by alkyl;

[0052] R6 are each independently selected from hydrogen, deuterium, halogen, cyano, cyclopropyl and C1-C6 alkyl optionally substituted by halogen or deuterium;

[0053] R7 is p is selected from 0, 1 and 2, R9 and R 10 are independently selected from hydrogen, C1-C3 alkyl, C1-C3 alkoxy and C3-C6 cycloalkyl, or R9 and R 10 Together with the nitrogen atom to which they are commonly attached, they form a 4-6 membered heterocyclic group, which is optionally substituted with one or more substituents selected from halogen, hydroxy and C1-C3 alkyl;

[0054] R8 is selected from hydrogen, deuterium, halogen, cyano and C1-C3 alkyl substituted by one or more halogens;

[0055] X is selected from oxygen atoms, -NH- and methylene, wherein the methylene is optionally substituted with one or more substituents selected from halogen, C3-C8 cycloalkyl, C3-C6 cycloalkylene and C1-C6 alkyl;

[0056] m is an integer from 1 to 3; and

[0057] n is an integer from 1 to 4.

[0058] In some embodiments, in the compound of formula (I),

[0059] R7 is p is 0 or 1, R9 is selected from hydrogen, C1-C3 alkyl, C1-C3 alkoxy and C3-C6 cycloalkyl, R10 is hydrogen, or R9 and R 10 Together with the nitrogen atom to which they are attached, they form a 4-6 membered heterocyclic group, which is optionally substituted with one or more halogens; and

[0060] R8 is selected from hydrogen, deuterium and halogen.

[0061] In some embodiments, in the compound of formula (I),

[0062] R1 is selected from hydrogen, deuterium, halogen, C1-C6 hydroxyalkyl, C1-C6 alkyl optionally substituted by halogen or deuterium, and C1-C6 alkoxy optionally substituted by halogen or deuterium;

[0063] R2 is each independently selected from hydrogen, deuterium, halogen, C1-C6 hydroxyalkyl, C1-C6 alkyl optionally substituted by halogen or deuterium, and C1-C6 alkoxy optionally substituted by halogen or deuterium;

[0064] R3 and R4 are each independently selected from hydrogen, halogen and C1-C4 alkyl optionally substituted by halogen, or R3 and R4 together with the carbon atom to which they are commonly attached form a C3-C6 cycloalkylene group optionally substituted by halogen, or R3 and R4 on adjacent carbon atoms together form a C3-C8 cycloalkane optionally substituted by halogen;

[0065] R5 is selected from C1-C6 alkyl optionally substituted by halogen or cyano, C3-C6 cycloalkyl optionally substituted by halogen or cyano, heterocyclyl optionally substituted by halogen or cyano, C1-C6 alkoxy optionally substituted by halogen or cyano, and amino optionally substituted by alkyl;

[0066] R6 are each independently selected from hydrogen, deuterium, halogen, cyano, cyclopropyl and C1-C6 alkyl optionally substituted by halogen or deuterium;

[0067] R7 is p is selected from 0, 1 and 2, R9 and R 10 are independently selected from hydrogen, C1-C3 alkyl, C1-C3 alkoxy and C3-C6 cycloalkyl, or R9 and R 10 Together with the nitrogen atom to which they are commonly attached, they form a 4-6 membered heterocyclic group, which is optionally substituted with one or more substituents selected from halogen, hydroxyl and C1-C3 alkyl; R' is selected from hydrogen, C1-C3 alkyl, C3-C6 cycloalkyl, aryl and heteroaryl;

[0068] R8 is selected from hydrogen, deuterium, halogen, cyano and C1-C3 alkyl substituted by one or more halogens;

[0069] X is selected from oxygen atoms, -NH- and methylene, wherein the methylene is optionally substituted with one or more substituents selected from halogen, C3-C8 cycloalkyl, C3-C6 cycloalkylene and C1-C6 alkyl;

[0070] m is an integer from 1 to 3; and

[0071] n is an integer from 1 to 4.

[0072] In some embodiments, in the compound of formula (I),

[0073] R7 is p is 0 or 1, R9 is selected from hydrogen, C1-C3 alkyl, C1-C3 alkoxy and C3-C6 cycloalkyl, R 10 is hydrogen, or R9 and R 10 Together with the nitrogen atom to which they are attached, they form a 4-6 membered heterocyclic group, which is optionally substituted with one or more halogens; R' is selected from hydrogen, C1-C3 alkyl, C3-C6 cycloalkyl, aryl and heteroaryl; and

[0074] R8 is selected from hydrogen, deuterium and halogen.

[0075] In some embodiments, in the compound of formula (I),

[0076] R1 is selected from hydrogen, deuterium, C1-C6 alkyl optionally substituted by halogen or deuterium, and halogen;

[0077] R2 are each independently selected from hydrogen, deuterium, C1-C6 alkyl optionally substituted by halogen or deuterium, and halogen;

[0078] R3 and R4 are each independently hydrogen or halogen, or R3 and R4 together with the carbon atom to which they are attached form a C3-C6 cycloalkylene group optionally substituted by halogen, or R3 and R4 on adjacent carbon atoms together form a C3-C8 cycloalkane optionally substituted by halogen;

[0079] R5 is selected from C1-C6 alkyl optionally substituted by halogen or cyano, C3-C6 cycloalkyl optionally substituted by halogen or cyano, heterocyclyl optionally substituted by halogen or cyano, C1-C6 alkoxy optionally substituted by halogen or cyano, and amino optionally substituted by alkyl;

[0080] R6 are each independently selected from hydrogen, deuterium, halogen, cyano, cyclopropyl and C1-C6 alkyl optionally substituted by halogen or deuterium;

[0081] R7 and R8 are each independently selected from:

[0082] a) hydrogen, deuterium, halogen, cyano, amino, hydroxy, C1-C6 alkyl optionally substituted by halogen, sulfone, sulfoxide, sulfonamide, sulfenamide, C 1-3and a C1-C6 alkoxy group optionally substituted by halogen;

[0083] b) p is selected from 0, 1 and 2, R9 and R 10 are independently selected from hydrogen, C1-C3 alkyl, C1-C3 alkoxy and C3-C6 cycloalkyl, or R9 and R 10 Together with the nitrogen atom to which they are commonly attached, they form a 4-6 membered heterocyclic group, which is optionally substituted with one or more substituents selected from halogen, hydroxyl and C1-C3 alkyl; R' is selected from hydrogen, C1-C3 alkyl, C3-C6 cycloalkyl, aryl and heteroaryl; and When p is 0, R9 and R 10 The combination of is not hydrogen and methyl;

[0084] c)

[0085] d) heterocyclyl and heteroaryl, each of which is optionally substituted with one or more substituents selected from oxo, halogen, hydroxy, carbonyl, C1-C6 alkyl and cyano, wherein the C1-C6 alkyl is optionally substituted with one or more halogen; and

[0086] e) Among them, R 11 is selected from C1-C6 alkyl, C1-C6 alkoxy, aryl, heteroaryl, C3-C8 cycloalkyl, heterocyclyl, C1-C6 cyanoalkyl, C3-C8 cycloalkyloxy and amino optionally substituted with C1-C6 alkyl; or

[0087] R7 and R8 together with the atoms to which they are attached form an optionally substituted aromatic or non-aromatic heterocyclic ring;

[0088] X is selected from oxygen atoms, -NH- and methylene, wherein the methylene is optionally substituted with one or more substituents selected from halogen, C3-C8 cycloalkyl, C3-C6 cycloalkylene and C1-C6 alkyl;

[0089] m is an integer from 1 to 3; and

[0090] n is an integer from 1 to 4.

[0091] In some embodiments, R5 is C1-C6 alkyl optionally substituted by halogen or cyano or C1-C6 alkoxy optionally substituted by halogen or cyano;

[0092] R6 are each independently selected from hydrogen, deuterium, halogen and cyano; and

[0093] n is an integer from 1 to 4.

[0094] In some embodiments, R1 is selected from hydrogen, deuterium, C1-C3 alkyl optionally substituted with halogen or deuterium, and halogen;

[0095] R2 are each independently selected from hydrogen, deuterium, C1-C3 alkyl optionally substituted by halogen or deuterium, and halogen;

[0096] R3 and R4 are each independently hydrogen or halogen, or R3 and R4 together with the carbon atom to which they are attached form a C3-C6 cycloalkylene group optionally substituted by halogen;

[0097] R5 is C1-C6 alkyl or C1-C6 alkoxy;

[0098] R6 are each independently selected from hydrogen, deuterium, halogen and cyano;

[0099] R7 and R8 are each independently selected from:

[0100] a) hydrogen, deuterium, halogen, cyano, amino, hydroxy, C1-C6 alkyl optionally substituted by halogen, sulfone, sulfoxide, sulfonamide, sulfenamide, C 1-3 and a C1-C6 alkoxy group optionally substituted by halogen;

[0101] b) p is selected from 0, 1 and 2, R9 and R 10 are independently selected from hydrogen, C1-C6 alkyl, C1-C6 alkoxy and C3-C8 cycloalkyl, or R9 and R 10 Together with the nitrogen atom to which they are commonly attached, they form a 4-6 membered heterocyclic group, wherein: the heterocyclic group is optionally substituted with one or more substituents selected from halogen, hydroxy, cyano, C1-C6 haloalkyl and C1-C6 alkyl; and When p is 0, R9 and R 10 The combination of is not hydrogen and methyl;

[0102] c)

[0103] d) heterocyclyl and heteroaryl, each of which is optionally substituted with one or more substituents selected from oxo, halogen, hydroxy, carbonyl, C1-C6 alkyl and cyano, wherein the C1-C6 alkyl is optionally substituted with one or more halogen; and

[0104] e) Among them, R 11 is selected from C1-C6 alkyl, C1-C6 alkoxy, aryl, heteroaryl, C3-C8 cycloalkyl, heterocyclyl, C1-C6 cyanoalkyl, C3-C8 cycloalkyloxy and amino optionally substituted with C1-C6 alkyl; or

[0105] R7 and R8 together with the atoms to which they are attached form an optionally substituted aromatic or non-aromatic heterocyclic ring;

[0106] m is an integer from 1 to 3; and

[0107] n is an integer from 1 to 4.

[0108] In some embodiments, in the compound of formula (I),

[0109] R1 is selected from hydrogen, deuterium, halogen, C1-C6 hydroxyalkyl, C1-C6 alkyl optionally substituted by halogen or deuterium, and C1-C6 alkoxy optionally substituted by halogen or deuterium;

[0110] R2 is each independently selected from hydrogen, deuterium, halogen, C1-C6 hydroxyalkyl, C1-C6 alkyl optionally substituted by halogen or deuterium, and C1-C6 alkoxy optionally substituted by halogen or deuterium;

[0111] R3 and R4 are each independently selected from hydrogen, halogen and C1-C4 alkyl optionally substituted by halogen, or R3 and R4 together with the carbon atom to which they are commonly attached form a C3-C6 cycloalkylene group optionally substituted by halogen, or R3 and R4 on adjacent carbon atoms together form a C3-C8 cycloalkane optionally substituted by halogen;

[0112] R5 is selected from C1-C6 alkyl optionally substituted by halogen or cyano, C3-C6 cycloalkyl optionally substituted by halogen or cyano, heterocyclyl optionally substituted by halogen or cyano, C1-C6 alkoxy optionally substituted by halogen or cyano, and amino optionally substituted by alkyl;

[0113] R6 are each independently selected from hydrogen, deuterium, halogen, cyano, cyclopropyl and C1-C6 alkyl optionally substituted by halogen or deuterium;

[0114] R7 is the following group optionally substituted by one or more substituents selected from a methyl group, a fluorine atom, a chlorine atom, a halomethyl group and a cyano group:

[0115]

[0116] R8 is selected from hydrogen, deuterium, halogen, cyano and C1-C3 alkyl substituted by one or more halogens;

[0117] X is selected from oxygen atoms, -NH- and methylene, wherein the methylene is optionally substituted with one or more substituents selected from halogen, C3-C8 cycloalkyl, C3-C6 cycloalkylene and C1-C6 alkyl;

[0118] m is an integer from 1 to 3; and

[0119] n is an integer from 1 to 4.

[0120] In some embodiments, in the compound of formula (I),

[0121] R1 is selected from hydrogen, deuterium, halogen, C1-C6 hydroxyalkyl, C1-C6 alkyl optionally substituted by halogen or deuterium, and C1-C6 alkoxy optionally substituted by halogen or deuterium;

[0122] R2 is each independently selected from hydrogen, deuterium, halogen, C1-C6 hydroxyalkyl, C1-C6 alkyl optionally substituted by halogen or deuterium, and C1-C6 alkoxy optionally substituted by halogen or deuterium;

[0123] R3 and R4 are each independently selected from hydrogen, halogen and C1-C4 alkyl optionally substituted by halogen, or R3 and R4 together with the carbon atom to which they are commonly attached form a C3-C6 cycloalkylene group optionally substituted by halogen, or R3 and R4 on adjacent carbon atoms together form a C3-C8 cycloalkane optionally substituted by halogen;

[0124] R5 is selected from C1-C6 alkyl optionally substituted by halogen or cyano, C3-C6 cycloalkyl optionally substituted by halogen or cyano, heterocyclyl optionally substituted by halogen or cyano, C1-C6 alkoxy optionally substituted by halogen or cyano, and amino optionally substituted by alkyl;

[0125] R7 and R8 together with the atoms to which they are attached form a heterocyclic ring A Heterocycle A is selected from the following structures:

[0126]

[0127] R6 are each independently selected from hydrogen, deuterium, halogen and cyano;

[0128] R 12 Each independently selected from halogen, C1-C3 alkyl and C3-C6 cycloalkylene, or adjacent R 12 Together they form a ring, said ring being optionally substituted with one or more halogen or C1-C3 alkyl;

[0129] m is an integer from 1 to 3;

[0130] n is an integer from 1 to 3; and

[0131] q is an integer from 0 to 6.

[0132] In some embodiments, in the compound of formula (I),

[0133] R1 is selected from hydrogen, deuterium, C1-C3 alkyl optionally substituted by halogen or deuterium, and halogen;

[0134] R2 are each independently selected from hydrogen, deuterium, C1-C3 alkyl optionally substituted by halogen or deuterium, and halogen;

[0135] R3 and R4 are each independently hydrogen or halogen, or R3 and R4 together with the carbon atom to which they are attached form a C3-C6 cycloalkylene group optionally substituted by halogen;

[0136] R5 is C1-C6 alkyl or C1-C6 alkoxy;

[0137] R7 and R8 together with the atoms to which they are attached form a heterocyclic ring A Heterocycle A is selected from the following structures:

[0138]

[0139] R6 are each independently selected from hydrogen, deuterium, halogen and cyano;

[0140] m is an integer from 1 to 3; and

[0141] n is an integer from 1 to 3.

[0142] In some embodiments, in the compound of formula (I),

[0143] R1 is selected from hydrogen, deuterium, halogen, C1-C6 hydroxyalkyl, C1-C6 alkyl optionally substituted by halogen or deuterium, and C1-C6 alkoxy optionally substituted by halogen or deuterium;

[0144] R2 is each independently selected from hydrogen, deuterium, halogen, C1-C6 hydroxyalkyl, C1-C6 alkyl optionally substituted by halogen or deuterium, and C1-C6 alkoxy optionally substituted by halogen or deuterium;

[0145] R3 and R4 are each independently selected from hydrogen, halogen and C1-C4 alkyl optionally substituted by halogen, or R3 and R4 together with the carbon atom to which they are commonly attached form a C3-C6 cycloalkylene group optionally substituted by halogen, or R3 and R4 on adjacent carbon atoms together form a C3-C8 cycloalkane optionally substituted by halogen;

[0146] R5 is selected from C1-C6 alkyl optionally substituted by halogen or cyano, C3-C6 cycloalkyl optionally substituted by halogen or cyano, heterocyclyl optionally substituted by halogen or cyano, C1-C6 alkoxy optionally substituted by halogen or cyano, and amino optionally substituted by alkyl;

[0147] R6 are each independently selected from hydrogen, deuterium, halogen, cyano, cyclopropyl and C1-C6 alkyl optionally substituted by halogen or deuterium;

[0148] R7 is selected from the following groups:

[0149]

[0150] R8 is selected from hydrogen, deuterium, halogen, cyano and C1-C3 alkyl substituted by one or more halogens;

[0151] X is selected from oxygen atoms, -NH- and methylene, wherein the methylene is optionally substituted with one or more substituents selected from halogen, C3-C8 cycloalkyl, C3-C6 cycloalkylene and C1-C6 alkyl;

[0152] m is an integer from 1 to 3; and

[0153] n is an integer from 1 to 4.

[0154] In some embodiments, in the compound of formula (I),

[0155] R1 is selected from hydrogen, deuterium, halogen, C1-C6 hydroxyalkyl, C1-C6 alkyl optionally substituted by halogen or deuterium, and C1-C6 alkoxy optionally substituted by halogen or deuterium;

[0156] R2 is each independently selected from hydrogen, deuterium, halogen, C1-C6 hydroxyalkyl, C1-C6 alkyl optionally substituted by halogen or deuterium, and C1-C6 alkoxy optionally substituted by halogen or deuterium;

[0157] R3 and R4 are each independently selected from hydrogen, halogen and C1-C4 alkyl optionally substituted by halogen, or R3 and R4 together with the carbon atom to which they are commonly attached form a C3-C6 cycloalkylene group optionally substituted by halogen, or R3 and R4 on adjacent carbon atoms together form a C3-C8 cycloalkane optionally substituted by halogen;

[0158] R5 is selected from C1-C6 alkyl optionally substituted by halogen or cyano, C3-C6 cycloalkyl optionally substituted by halogen or cyano, heterocyclyl optionally substituted by halogen or cyano, C1-C6 alkoxy optionally substituted by halogen or cyano, and amino optionally substituted by alkyl;

[0159] R6 are each independently selected from hydrogen, deuterium, halogen, cyano, cyclopropyl and C1-C6 alkyl optionally substituted by halogen or deuterium;

[0160] R7 is selected from the following groups:

[0161]

[0162]

[0163] R8 is selected from hydrogen, deuterium, halogen, cyano and C1-C3 alkyl substituted by one or more halogens;

[0164] X is selected from oxygen atoms, -NH- and methylene, wherein the methylene is optionally substituted with one or more substituents selected from halogen, C3-C8 cycloalkyl, C3-C6 cycloalkylene and C1-C6 alkyl;

[0165] m is an integer from 1 to 3; and

[0166] n is an integer from 1 to 4.

[0167] The present disclosure also provides a compound of formula (I-1), or a pharmaceutically acceptable salt thereof, or an isomer thereof,

[0168]

[0169] wherein R1 is selected from hydrogen, deuterium, C1-C3 alkyl optionally substituted by halogen or deuterium, and halogen;

[0170] R2 are each independently selected from hydrogen, deuterium, C1-C3 alkyl optionally substituted by halogen or deuterium, and halogen;

[0171] R5 is C1-C3 alkyl or C1-C3 alkoxy;

[0172] R 6a and R 6b are each independently selected from hydrogen, deuterium, a chlorine atom, a fluorine atom and a cyano group;

[0173] R7 is a 4-6 membered heterocyclic group, wherein the heterocyclic group comprises -NH-C(=O)- or -NH-S(=O)2-, wherein the heterocyclic group is optionally substituted by one or more substituents selected from oxo, halogen, hydroxyl, carbonyl, C1-C3 alkyl and cyano, wherein the C1-C3 alkyl is optionally substituted by one or more halogen;

[0174] R8 is selected from hydrogen, deuterium, halogen, cyano and C1-C3 alkyl substituted by one or more halogens; and

[0175] m is an integer from 1 to 3.

[0176] In some embodiments, R7 in the compound of formula (I-1) is the following group optionally substituted by one or more substituents selected from methyl, fluorine, chlorine, halomethyl and cyano:

[0177]

[0178]

[0179] and

[0180] R8 is selected from hydrogen, deuterium, halogen and cyano.

[0181] The present disclosure also provides a compound of formula (I-1), or a pharmaceutically acceptable salt thereof, or an isomer thereof,

[0182]

[0183] wherein R1 is selected from hydrogen, deuterium, C1-C3 alkyl optionally substituted by halogen or deuterium, and halogen;

[0184] R2 are each independently selected from hydrogen, deuterium, C1-C3 alkyl optionally substituted by halogen or deuterium, and halogen;

[0185] R5 is C1-C3 alkyl or C1-C3 alkoxy;

[0186] R 6a and R 6b are each independently selected from hydrogen, deuterium, a chlorine atom, a fluorine atom and a cyano group;

[0187] R7 and R8 together with the atoms to which they are attached form a 4-8 membered non-aromatic heterocyclic ring, said heterocyclic ring being monocyclic or bicyclic, said heterocyclic ring comprising -NH-C(=O)- or -NH-S(=O)2-, said heterocyclic ring being optionally substituted with one or more substituents selected from C1-C3 alkylamide, halogen, oxo, C1-C3 alkyl optionally substituted with halogen, and C1-C3 alkoxy; and

[0188] m is an integer from 1 to 3.

[0189] In some embodiments, in the compound of formula (I-1), R7 and R8 together with the atoms to which they are attached form a heterocyclic ring A. Heterocycle A is selected from the following structures:

[0190]

[0191]

[0192] R 6a and R 6b are each independently selected from hydrogen, deuterium, chlorine atoms and fluorine atoms;

[0193] R 12 Each independently selected from halogen, C1-C3 alkyl and C3-C6 cycloalkylene, or adjacent R 12 Together they form a ring, said ring being optionally substituted with one or more halogen or C1-C3 alkyl; and

[0194] q is an integer from 0 to 6.

[0195] In some embodiments, in the compound of formula (I-1), R7 and R8 together with the atoms to which they are attached form a heterocyclic ring A. Heterocycle A is selected from the following structures:

[0196]

[0197] and

[0198] R 6a and R 6b are each independently selected from hydrogen, deuterium, a chlorine atom and a fluorine atom.

[0199] The present disclosure also provides a compound of formula (I-1), or a pharmaceutically acceptable salt thereof, or an isomer thereof,

[0200]

[0201] wherein R1 is selected from hydrogen, deuterium, C1-C3 alkyl optionally substituted by halogen or deuterium, and halogen;

[0202] R2 are each independently selected from hydrogen, deuterium, C1-C3 alkyl optionally substituted by halogen or deuterium, and halogen;

[0203] R5 is C1-C3 alkyl or C1-C3 alkoxy;

[0204] R 6a and R 6b are each independently selected from hydrogen, deuterium, a chlorine atom, a fluorine atom and a cyano group;

[0205] R7 is p is selected from 0, 1 and 2, R9 and R 10 are independently selected from hydrogen, C1-C3 alkyl, C1-C3 alkoxy and C3-C6 cycloalkyl, or R9 and R 10 Together with the nitrogen atom to which they are commonly attached, they form a 4-6 membered heterocyclic group, which is optionally substituted with one or more substituents selected from halogen, hydroxy and C1-C3 alkyl;

[0206] R8 is selected from hydrogen, deuterium, halogen, cyano and C1-C3 alkyl substituted by one or more halogens; and

[0207] m is an integer from 1 to 3.

[0208] In some embodiments, R7 in the compound of formula (I-1) is selected from the following groups:

[0209]

[0210] and

[0211] R8 is selected from hydrogen, deuterium, halogen, cyano and C1-C3 alkyl substituted by one or more halogens.

[0212] The present disclosure also provides a compound of formula (I-1), or a pharmaceutically acceptable salt thereof, or an isomer thereof,

[0213]

[0214] wherein R1 is selected from hydrogen, deuterium, C1-C3 alkyl optionally substituted by halogen or deuterium, and halogen;

[0215] R2 are each independently selected from hydrogen, deuterium, C1-C3 alkyl optionally substituted by halogen or deuterium, and halogen;

[0216] R5 is C1-C3 alkyl or C1-C3 alkoxy;

[0217] R 6a and R 6b are each independently selected from hydrogen, deuterium, a chlorine atom, a fluorine atom and a cyano group;

[0218] R7 is p is selected from 0, 1 and 2, R9 and R 10 are independently selected from hydrogen, C1-C3 alkyl, C1-C3 alkoxy and C3-C6 cycloalkyl, or R9 and R 10 Together with the nitrogen atom to which they are commonly attached, they form a 4-6 membered heterocyclic group, which is optionally substituted with one or more substituents selected from halogen, hydroxyl and C1-C3 alkyl; R' is selected from hydrogen, C1-C3 alkyl, C3-C6 cycloalkyl, aryl and heteroaryl;

[0219] R8 is selected from hydrogen, deuterium, halogen, cyano and C1-C3 alkyl substituted by one or more halogens; and

[0220] m is an integer from 1 to 3.

[0221] In some embodiments, R7 in the compound of formula (I-1) is selected from the following groups:

[0222]

[0223] and

[0224] R8 is selected from hydrogen, deuterium, halogen, cyano and C1-C3 alkyl substituted by one or more halogens.

[0225] The present disclosure also provides the following compounds, or pharmaceutically acceptable salts thereof, or isomers thereof:

[0226]

[0227]

[0228] The present disclosure provides a method for preparing a compound of formula (I) or a pharmaceutically acceptable salt thereof, or an isomer thereof, comprising the following steps:

[0229]

[0230] The compound represented by formula (Ia) reacts with the compound represented by formula (Ib) under alkaline conditions to obtain the compound represented by formula (Ic); the compound represented by formula (Ic) undergoes a reduction reaction to obtain the compound represented by formula (Id); the compound represented by formula (Id) and the compound represented by formula (Ie) undergo a ring-closure reaction under acidic conditions to obtain the compound represented by formula (I);

[0231] R1, R2, R3, R4, R5, R6, R7, R8, X, m and n are as defined for the compound of formula (I); and Y is selected from halogen, sulfonyl and sulfinyl.

[0232] The present disclosure also provides another method for preparing a compound of formula (I) or a pharmaceutically acceptable salt thereof, or an isomer thereof, comprising the following steps:

[0233]

[0234] The compound represented by formula (Ia) reacts with the compound represented by formula (Ib) under alkaline conditions to obtain the compound represented by formula (Ic); the compound represented by formula (Ic) is subjected to a reduction reaction to obtain the compound represented by formula (Id); the compound represented by formula (Id) and the compound represented by formula (If) are subjected to a ring-closure reaction under acidic conditions to obtain the compound represented by formula (Ig); the compound represented by formula (Ig) is reacted in the presence of a catalyst to obtain the compound represented by formula (I);

[0235] The catalyst is selected from palladium / carbon, Raney nickel, tetrakistriphenylphosphine palladium, palladium dichloride, palladium acetate, [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium, 1,1'-bis(dibenzylphosphino)dichlorodipentaferropalladium, tris(dibenzylideneacetone)dipalladium, 2-dicyclohexylphosphine-2',6'-dimethoxybiphenyl, [1,1'-bis(di-tert-butylphosphino)ferrocene]dichloropalladium(II), cuprous iodide, cuprous bromide, cuprous chloride and copper trifluoromethanesulfonate;

[0236] R1, R2, R3, R4, R5, R6, R7, R8, X, m and n are as defined in the compound of Formula I; and

[0237] Y and Z are each independently selected from halogen, sulfonyl and sulfinyl.

[0238] The present disclosure also provides a method for preparing the compound or its pharmaceutically acceptable salt, or its isomer, in particular, by using the method of the examples.

[0239] The present disclosure also relates to a pharmaceutical composition comprising the compound described herein or a pharmaceutically acceptable salt thereof, or an isomer thereof. Generally, the composition further comprises at least one pharmaceutically acceptable carrier, diluent, or excipient.

[0240] In some embodiments, in the pharmaceutical composition, the unit dose of the compound or its pharmaceutically acceptable salt, or its isomer is 0.001 mg-1000 mg.

[0241] In certain embodiments, the pharmaceutical composition contains 0.01-99.99% of the aforementioned compound or a pharmaceutically acceptable salt thereof, based on the total weight of the composition. In certain embodiments, the pharmaceutical composition contains 0.1-99.9% of the aforementioned compound or a pharmaceutically acceptable salt thereof. In certain embodiments, the pharmaceutical composition contains 0.5%-99.5% of the aforementioned compound or a pharmaceutically acceptable salt thereof. In certain embodiments, the pharmaceutical composition contains 1%-99% of the aforementioned compound or a pharmaceutically acceptable salt thereof. In certain embodiments, the pharmaceutical composition contains 2%-98% of the aforementioned compound or a pharmaceutically acceptable salt thereof.

[0242] In certain embodiments, the pharmaceutical composition comprises 0.01% to 99.99% of a pharmaceutically acceptable excipient, based on the total weight of the composition. In certain embodiments, the pharmaceutical composition comprises 0.1% to 99.9% of a pharmaceutically acceptable excipient. In certain embodiments, the pharmaceutical composition comprises 0.5% to 99.5% of a pharmaceutically acceptable excipient. In certain embodiments, the pharmaceutical composition comprises 1% to 99% of a pharmaceutically acceptable excipient. In certain embodiments, the pharmaceutical composition comprises 2% to 98% of a pharmaceutically acceptable excipient.

[0243] The present disclosure also relates to the use of the compound or its pharmaceutically acceptable salt, or its isomer, or a pharmaceutical composition comprising the same in the preparation of a drug for treating diseases related to P2X3 activity.

[0244] The present disclosure also relates to the compound or its pharmaceutically acceptable salt, or its isomer, or a pharmaceutical composition comprising the same, for use as a medicament.

[0245] The present disclosure also relates to the compound or its pharmaceutically acceptable salt, or its isomer, or a pharmaceutical composition comprising the same, for treating diseases related to P2X3 activity.

[0246] The present disclosure also relates to a method for treating a disease associated with P2X3 activity, comprising administering a therapeutically effective amount of the compound or its pharmaceutically acceptable salt, or its isomer, or a pharmaceutical composition comprising the same to a patient in need thereof.

[0247] In some embodiments, the disease associated with P2X3 activity refers to a disease associated with excessive P2X3 activity. The compounds of the present disclosure are highly selective for P2X3 and can prevent taste loss. In some embodiments, the compounds of the present disclosure have an antagonistic effect on P2X3 homoreceptors that is more than 20 times stronger than the antagonistic effect on P2X2 / 3 heteromeric receptors. In some embodiments, the compounds of the present disclosure have an antagonistic effect on P2X3 homoreceptors that is more than 30 times stronger than the antagonistic effect on P2X2 / 3 heteromeric receptors. In some embodiments, the compounds of the present disclosure have an antagonistic effect on P2X3 homoreceptors that is more than 50 times stronger than the antagonistic effect on P2X2 / 3 heteromeric receptors. In some embodiments, the compounds of the present disclosure have an antagonistic effect on P2X3 homoreceptors that is more than 100 times stronger than the antagonistic effect on P2X2 / 3 heteromeric receptors.

[0248] The present disclosure also relates to the use of the compound or its pharmaceutically acceptable salt, or its isomer, or a pharmaceutical composition containing the same in the preparation of drugs for treating pain, urinary tract diseases, cough and the like.

[0249] The present disclosure also relates to the compound or its pharmaceutically acceptable salt, or its isomer, or a pharmaceutical composition comprising the same, which is used for treating pain, urinary tract diseases, cough and the like.

[0250] The present disclosure also relates to a method for treating pain, urinary tract diseases, cough and the like, which comprises administering to a patient in need thereof a therapeutically effective amount of the compound or its pharmaceutically acceptable salt, or its isomer, or a pharmaceutical composition comprising the same.

[0251] In some embodiments, the pain can be, for example, chronic pain, neuropathic pain, acute pain, back pain, cancer pain, pain caused by rheumatoid arthritis, migraines, and visceral pain. Urinary tract disorders, such as overactive bladder (also known as urinary incontinence), pelvic hypersensitivity, and urethritis.

[0252] In some embodiments, the compounds of the present disclosure, or pharmaceutically acceptable salts thereof, or isomers thereof, or pharmaceutical compositions comprising the same, can be used to treat gastrointestinal disorders, including, for example, constipation and functional gastrointestinal disorders (e.g., irritable bowel syndrome or functional dyspepsia); can be used to treat cancer; can be used to treat cardiovascular disorders or for cardioprotection after myocardial infarction; can be used as immunomodulators, particularly for the treatment of autoimmune diseases (e.g., arthritis), for skin transplants, organ transplants, or similar surgical needs, for collagen diseases, for allergies, or as antitumor agents or antiviral agents; can be used to treat multiple sclerosis, Parkinson's disease, and Huntington's chorea; can be used to treat depression, anxiety, stress-related disorders (e.g., post-traumatic stress disorder, panic disorder, social phobia, or obsessive-compulsive disorder), premature ejaculation, psychosis, traumatic brain injury, stroke, Alzheimer's disease, disease), spinal cord injury, drug addiction (e.g., treatment of alcohol, nicotine, opioid, or other drug abuse), or sympathetic nervous system disorders (e.g., high blood pressure); can be used to treat diarrhea; can be used to treat lung disorders, such as asthma, cough, or pulmonary edema.

[0253] The compounds of the present disclosure, or pharmaceutically acceptable salts thereof, or isomers thereof, can be formulated for administration orally, buccally, vaginally, rectally, by inhalation, by insufflation, intranasally, sublingually, topically, or parenterally (e.g., intramuscularly, subcutaneously, intraperitoneally, intrathoracically, intravenously, epidurally, intrathecally, intracerebroventricularly, or by injection into a joint).

[0254] The pharmaceutically acceptable salts of the compounds described in the present disclosure may be selected from inorganic salts or organic salts.

[0255] The term "treating" refers to the administration of a pharmaceutical composition for prophylactic and / or therapeutic purposes. For "preventing a disease," this refers to the prophylactic treatment of a subject who is not already suffering from the disease but is susceptible to or at risk for the disease. For "treating a disease," this refers to the treatment of a patient already suffering from the disease to improve or stabilize the patient's condition.

[0256] Any isotopically labeled (or "radiolabeled") derivative of the compounds of the present disclosure, or pharmaceutically acceptable salts thereof, or isomers thereof, is covered by the present disclosure. Such derivatives are those in which one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of radionuclides that may be incorporated include 2 H (also written as "D" for deuterium), 3 H (also written as "T" for tritium), 11 C. 13 C. 14 C.13 N. 15 N. 15 O. 17 O. 18 O. 18 F. 36 Cl, 82 Br, 75 Br, 76 Br, 77 Br, 123 I. 124 I. 125 I. 31 P. 32 P. 35 S, and 131 I. The radionuclide used will depend on the specific application of the radiolabeled derivative. For example, for in vitro receptor labeling and competition assays, 3 H or 14 C is often useful. For radiographic applications, 11 C or 18 F is often useful. In some embodiments, the radionuclide is 3 H. In some embodiments, the radionuclide is 14 C. In some embodiments, the radionuclide is 11 C. And in some embodiments, the radionuclide is 18 F.

[0257] Unless otherwise noted, when a position is specifically designated as deuterium (D), the position is understood to have an abundance of deuterium at least 3000 times greater than the natural abundance of deuterium, which is 0.015% (ie, at least 45% deuterium incorporation).

[0258] Unless stated otherwise, the following terms used in the specification and claims have the following meanings.

[0259] A "pharmaceutical composition" refers to a mixture containing one or more compounds described herein, or their physiologically acceptable salts or prodrugs, together with other chemical components, as well as other components such as physiologically acceptable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate administration to an organism, facilitating absorption of the active ingredients and thereby exerting their biological activity.

[0260] "Pharmaceutically acceptable excipients" include, but are not limited to, any adjuvant, carrier, excipient, glidant, sweetener, diluent, preservative, dye / colorant, flavoring agent, surfactant, wetting agent, dispersant, suspending agent, stabilizer, isotonic agent, solvent or emulsifier approved by the U.S. Food and Drug Administration for use in humans or domestic animals.

[0261] As used herein, an "effective amount" or "therapeutically effective amount" encompasses an amount sufficient to ameliorate or prevent the symptoms or conditions of a medical condition. An effective amount also refers to an amount sufficient to permit or facilitate diagnosis. The effective amount for a particular patient or veterinary subject may vary depending on factors such as the condition to be treated, the patient's overall health, the route and dosage of administration, and the severity of side effects. An effective amount can be the maximum dose or dosage regimen that avoids significant side effects or toxic effects.

[0262] In the chemical structures of the compounds disclosed herein, the bond Indicates that the configuration is not specified, that is, if chiral isomers exist in the chemical structure, the bond Can be or include both Two configurations. In the chemical structure of the compound disclosed in the present invention, the bond No configuration is specified, i.e., the bond The configuration can be E-type or Z-type, or include both E and Z configurations.

[0263] Compounds and intermediates of the present disclosure can also exist in different tautomeric forms, and all such forms are included in the scope of the present disclosure. The term "tautomer" or "tautomeric form" refers to structural isomers of different energies that can interconvert via a low energy barrier. For example, proton tautomers (also referred to as prototransfer tautomers) include interconversions via proton migration, such as keto-enol and imine-enamine, lactam-lactim isomerization. The lactam-lactim equilibrium example is between A and B as shown below.

[0264]

[0265] "Halogen" refers to fluorine, chlorine, bromine and iodine.

[0266] All compounds in this disclosure can be drawn as either Form A or Form B. All tautomeric forms are within the scope of this disclosure. The naming of the compounds does not exclude any tautomers.

[0267] "Alkyl" refers to a straight or branched chain alkyl group, including straight and branched groups of 1 to 20 carbon atoms. Preferably, it contains 1 to 6 carbon atoms, more preferably 1 to 3 carbon atoms, such as methyl, ethyl, n-propyl, isopropyl, etc. The alkyl group can be substituted or unsubstituted. When substituted, the substituent can be substituted at any available point of attachment, preferably one or more of the following groups, independently selected from halogen, hydroxy, oxo, amino, C 1-6 Alkyl, C 1-6Alkoxy, 3 to 6 membered cycloalkyl, 3 to 6 membered heterocycloalkyl, 6 to 10 membered aryl or 5 to 10 membered heteroaryl, wherein the alkyl, alkoxy, cycloalkyl, heterocycloalkyl, aryl or heteroaryl group is optionally substituted by one or more halogen, hydroxyl, amino, C 1-6 Alkyl or C 1-6 Alkoxy is substituted. "Alkoxy" refers to an alkyloxy group, wherein the alkyl group is as defined above, for example, methoxy, ethoxy, etc. Alkoxy groups may be optionally substituted or unsubstituted, and when substituted, the substituents are preferably one or more of the following groups, which are independently selected from halogen, hydroxy, oxo, nitro, cyano, amino, C 1-6 Alkyl, C 1-6 Alkoxy, 3 to 6 membered cycloalkyl, 3 to 6 membered heterocycloalkyl, said alkyl, alkoxy, cycloalkyl or heterocycloalkyl being substituted by one or more selected from halogen, hydroxy, amino, C 1-6 Alkyl or C 1-6 Alkoxy substitution.

[0268] "Heterocyclyl" refers to a non-aromatic ring group containing 1 to 6 heteroatoms and 3 to 18 ring atoms, wherein the heteroatoms are selected from oxygen, nitrogen and sulfur. The heterocyclyl preferably contains 1 to 4 heteroatoms, more preferably 1 to 3 heteroatoms, and even more preferably 1 or 2 heteroatoms; the heterocyclyl is preferably 3 to 12 members, more preferably 3 to 8 members or 4 to 8 members, even more preferably 4 to 6 members, and even more preferably 5 or 6 members. Unless otherwise specified in this specification, the heterocyclyl can be a monocyclic, bicyclic, tricyclic or tetracyclic ring system, which can include a spirocyclic or bridged ring system; and the nitrogen, carbon or sulfur atoms in the heterocyclyl can be optionally oxidized; the nitrogen atom can be optionally quaternized; and the heterocyclyl can be partially or fully saturated. The "heterocyclic group containing -NH-C(=O)- or -NH-S(=O)2-" ring structure may contain heteroatoms in addition to the "-NH-C(=O)-" and "-NH-S(=O)2-" parts.

[0269] The term "aryl" refers to a 6- to 14-membered all-carbon monocyclic or fused polycyclic (fused polycyclic is a ring which shares adjacent pairs of carbon atoms) group having a conjugated pi electron system, preferably 6- to 10-membered, such as phenyl and naphthyl.

[0270] "Heteroaryl" refers to an aromatic ring group containing 1 to 4 heteroatoms and 5 to 14 ring atoms, wherein the heteroatoms are selected from oxygen, sulfur and nitrogen. Heteroaryl is preferably 4 to 6-membered or 6 to 12-membered, more preferably 5-membered or 6-membered. For example. Non-limiting examples include: imidazolyl, furyl, thienyl, thiazolyl, pyrazolyl, oxazolyl, pyrrolyl, tetrazolyl, pyridyl, pyrimidinyl, thiadiazole, pyrazine, etc.

[0271] The heteroaryl group may be optionally substituted or unsubstituted, and when substituted, the substituents are preferably one or more groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, carboxyl or carboxylate.

[0272] Aryl or heteroaryl may be optionally substituted or unsubstituted. When substituted, the substituents are preferably one or more of the following groups independently selected from halogen, hydroxy, oxo, nitro, cyano, amino, C 1-6 Alkyl, C 1-6 Alkoxy, 3 to 6 membered cycloalkyl, 3 to 6 membered heterocycloalkyl, said alkyl, alkoxy, cycloalkyl or heterocycloalkyl being substituted by one or more selected from halogen, hydroxy, amino, C 1-6 Alkyl or C 1-6 Alkoxy substitution.

[0273] "Cycloalkyl" or "cycloalkyl" refers to a stable, non-aromatic monocyclic or polycyclic alkyl group consisting solely of carbon and hydrogen atoms, which may contain spiro or bridged ring systems, having 3 to 15 carbon atoms, 3 to 10 carbon atoms, 3 to 8 carbon atoms, 3 to 6 carbon atoms, or 5 to 7 carbon atoms, and which is saturated or unsaturated and is attached to the rest of the molecule by a single bond. Monocyclic cycloalkyl groups include non-bridged cycloalkyl groups such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. Cycloalkyl groups may be substituted or unsubstituted, and when substituted, the substituents are preferably one or more of the following groups independently selected from halogen, hydroxy, oxo, nitro, cyano, amino, C 1-6 Alkyl, C 1-6 Alkoxy, 3 to 6 membered cycloalkyl, 3 to 6 membered heterocycloalkyl, said alkyl, alkoxy, cycloalkyl or heterocycloalkyl being substituted by one or more selected from halogen, hydroxy, amino, C 1-6 Alkyl or C 1-6 Alkoxy substituted. "Cycloalkylene" or "cycloalkylene" refers to a divalent cyclic hydrocarbon radical derived from a cycloalkyl radical. For example, It may be substituted or unsubstituted. When substituted, the substituent is preferably one or more of the following groups, which are independently selected from halogen, hydroxy, oxo, nitro, cyano, amino, C 1-6 Alkyl, C 1-6 Alkoxy, 3 to 6 membered cycloalkyl, 3 to 6 membered heterocycloalkyl, said alkyl, alkoxy, cycloalkyl or heterocycloalkyl being substituted by one or more selected from halogen, hydroxy, amino, C 1-6 Alkyl or C 1-6 Alkoxy substitution.

[0274] "Optionally" or "optionally" means that the subsequently described event or circumstance may but need not occur, and the description includes instances where the event or circumstance occurs and instances where it does not. For example, "C1-C6 alkyl optionally substituted with halogen or cyano" means that halogen or cyano may but need not be present, and the description includes instances where the alkyl group is substituted with halogen or cyano and instances where the alkyl group is not substituted with halogen or cyano.

[0275] The term "hydroxy" refers to an -OH group.

[0276] The term "halogen" refers to fluorine, chlorine, bromine or iodine.

[0277] The term "cyano" refers to -CN.

[0278] The term "amino" refers to -NH2.

[0279] The term "nitro" refers to -NO2.

[0280] The term "oxo" refers to a =0 substituent.

[0281] The term "carboxy" refers to -C(O)OH.

[0282] "Substituted" means that one or more hydrogen atoms, preferably 1 to 5, more preferably 1 to 3 hydrogen atoms in the group are independently replaced by a corresponding number of substituents.

[0283] "Pharmaceutically acceptable salts" refer to salts of the disclosed compounds that are safe and effective for use in mammals and possess the desired biological activity. Salts can be prepared during the final isolation and purification of the compounds, or separately by reacting a suitable group with a suitable base or acid. Bases commonly used to form pharmaceutically acceptable salts include inorganic bases, such as sodium hydroxide and potassium hydroxide, and organic bases, such as ammonia. Acids commonly used to form pharmaceutically acceptable salts include inorganic acids and organic acids. DETAILED DESCRIPTION

[0284] The structures of the compounds were determined by nuclear magnetic resonance (NMR) and / or mass spectrometry (MS). -6 The unit of ppm is given. NMR measurements were performed using a Bruker AVANCE-400 NMR spectrometer. The solvents used were deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), and deuterated methanol (CD3OD), and tetramethylsilane (TMS) was used as the internal standard.

[0285] HPLC determinations were performed using a Waters ACQUITY ultra high performance LC, Shimadzu LC-20A systems, Shimadzu LC-2010HT series, or Agilent 1200 LC high pressure liquid chromatograph (ACQUITY UPLC BEH C18 1.7 μm 2.1*50 mm column, Ultimate XB-C18 3.0*150 mm column, or Xtimate C18 2.1*30 mm column).

[0286] MS was measured using a Waters SQD2 mass spectrometer in positive / negative ion mode with a mass scan range of 100-1200.

[0287] Chiral HPLC analysis was performed using Chiralpak IC-3 100×4.6mm ID, 3μm, Chiralpak AD-3 150×4.6mm ID, 3μm, Chiralpak AD-3 50×4.6mm ID, 3μm, Chiralpak AS-3 150×4.6mm ID, 3μm, Chiralpak AS-3 100×4.6mm ID, 3μm, ChiralCel OD-3 150×4.6mm ID, 3μm, ChiralCel OD-3 100×4.6mm ID, 3μm, ChiralCel OJ-H 150×4.6mm ID, 5μm, and ChiralCel OJ-3 150×4.6mm ID, 3μm columns.

[0288] The thin layer chromatography silica gel plate uses Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plate. The specification of the silica gel plate used in thin layer chromatography (TLC) is 0.15mm~0.2mm, and the specification used for thin layer chromatography separation and purification products is 0.4mm~0.5mm.

[0289] Column chromatography generally uses Yantai Huanghai silica gel 100-200 mesh, 200-300 mesh or 300-400 mesh silica gel as the carrier.

[0290] Chiral preparative columns used were DAICEL CHIRALPAK IC (250 mm*30 mm, 10 μm) or Phenomenex-Amylose-1 (250 mm*30 mm, 5 μm).

[0291] The known starting materials disclosed herein can be synthesized by methods known in the art, or can be purchased from ABCR GmbH & Co. KG, Acros Organics, Aldrich Chemical Company, AccelaChemBio Inc, Darui Chemicals, and other companies.

[0292] Unless otherwise specified in the examples, all reactions can be carried out under argon or nitrogen atmosphere.

[0293] Argon atmosphere or nitrogen atmosphere means that the reaction bottle is connected to an argon or nitrogen balloon with a volume of about 1L.

[0294] Hydrogen atmosphere means that the reaction bottle is connected to a hydrogen balloon with a capacity of about 1L.

[0295] The pressurized hydrogenation reaction uses a Parr 3916EKX hydrogenator and a Qinglan QL-500 hydrogen generator or an HC2-SS hydrogenator.

[0296] The hydrogenation reaction is usually carried out by evacuating the chamber and filling it with hydrogen, and the operation is repeated three times.

[0297] Unless otherwise specified in the examples, the solution refers to an aqueous solution.

[0298] Unless otherwise specified in the examples, the reaction temperature is room temperature, 20°C to 30°C.

[0299] The reaction progress in the examples was monitored by thin layer chromatography (TLC). The developing solvent used in the reaction, the column chromatography eluent system used to purify the compound, and the developing solvent system for thin layer chromatography, the volume ratio of the solvent were adjusted according to the polarity of the compound, and a small amount of alkaline or acidic reagents such as triethylamine and acetic acid could be added for adjustment.

[0300] The preparation of the positive compound MK-7264 was based on the experimental steps of patent WO2005095359.

[0301] Example 1

[0302] 1-[4-(1-{[(2R)-4-acetylmorpholin-2-yl]methyl}-5-methyl-1H-1,3-benzodiazol-2-yl)-3,5-difluorophenyl]pyrrolidin-2-one (1)

[0303]

[0304] Step 1: Tert-butyl (2S)-2-[(1,3-dicarbonyl-2,3-dihydro-1H-isoindol-2-yl)methyl]morpholine-4-carboxylate (1a)

[0305] Tert-butyl (2R)-2-(hydroxymethyl)morpholine-4-carboxylate (3.00 g, 13.81 mmol), 2,3-dihydro-1H-isoindole-1,3-dione (2.23 g, 15.19 mmol), and triphenylphosphine (5.43 g, 20.71 mmol) were dissolved in THF (50 mL). Diisopropyl azodicarboxylate (4.1 mL, 20.71 mmol) was added dropwise at 0°C under nitrogen. After the reaction mixture was stirred at room temperature for 12 hours, LCMS indicated complete reaction of the starting material. The mixture was concentrated under reduced pressure, and the residue was purified by flash silica gel chromatography (ethyl acetate / petroleum ether = 0-30%, 60 mL / min) to afford the title compound 1a (6.00 g, 87.8% yield).

[0306] MS (ESI) m / z = 291.1 [M+H] + .

[0307] 1 H NMR (400MHz, CDCl3) δ7.83-7.76(m,2H),7.69-7.63(m,2H),4.91(spt,J=6.2Hz,3H),3.96-3.84(m,1H),3 .83-3.77(m,2H),3.75-3.64(m,2H),3.60(dd,J=4.6,13.6Hz,1H),3.37(dt,J=2.9,11.4Hz,1H),2.92(br s,1H),2.69(br s,1H),1.39(s,9H),1.21(s,11H).

[0308] Step 2: tert-Butyl (2S)-2-(aminomethyl)morpholine-4-carboxylate (1b)

[0309] To a solution of compound 1a (6.00 g, 12.12 mmol) in ethanol (150 mL) was added hydrazine hydrate (1.07 g, 18.19 mmol) at room temperature. The reaction mixture was then stirred at 80°C for 1 hour. LCMS indicated the reaction was complete. The mixture was cooled to room temperature, filtered, and the filtrate was concentrated under reduced pressure. The residue was dissolved in ethyl acetate (80 mL) and filtered. The filtrate was concentrated under reduced pressure to afford the title compound 1b (3.00 g, 91.5% yield).

[0310] 1H NMR (400MHz, CDCl3) δ6.27(br s,2H),3.82(br d,J=10.9Hz,3H),3.51-3.41(m,1H),3.34-3.23(m,1H),3.29(br dd,J=3.7,6.7Hz,1H),2.85(br s,1H),2.73-2.63(m,2H),2.57(br s,1H),1.40(s,9H).

[0311] Step 3: tert-Butyl (2S)-2-{[(4-methyl-2-nitrophenyl)amino]methyl}morpholine-4-carboxylate (1c)

[0312] To a solution of compound 1b (3.00 g, 11.10 mmol) in 1,4-dioxane (60 mL) at room temperature were added 1-fluoro-4-methyl-2-nitrobenzene (1.72 g, 11.10 mmol) and ethyldiisopropylamine (3.7 mL, 22.19 mmol). The reaction mixture was then stirred at 100°C for 12 hours. LCMS indicated complete disappearance of the starting material. Water (50 mL) was added, and the mixture was extracted with ethyl acetate (80 mL x 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to afford the crude product, which was then purified by flash silica gel chromatography (ethyl acetate / petroleum ether = 0-17%, 60 mL / min) to afford the title compound 1c (2.35 g, 60% yield).

[0313] MS (ESI) m / z = 374.2 [M + Na] + .

[0314] 1 H NMR(400MHz, CDCl3)δ8.11(br s,1H),7.99(d,J=0.8Hz,1H),7.30-7.27(m,1H),6.77(d,J=8.8Hz,1H),4.06-3.81(m,3 H),3.75-3.65(m,1H),3.64-3.53(m,1H),3.47-3.29(m,2H),3.09-2.91(m,1H),2.80(br s,1H),2.28(s,3H),1.47(s,9H).

[0315] Step 4: tert-Butyl (2S)-2-{[(2-amino-4-methylphenyl)amino]methyl}morpholine-4-carboxylate (1d)

[0316] To a solution of compound 1c (2.00 g, 5.69 mmol) in methanol (20 mL) was added 10% Pd / C (100 mg) at room temperature. The resulting mixture was purged with hydrogen three times, and the reaction mixture was stirred at room temperature for 12 hours. LCMS showed complete disappearance of the starting material. The mixture was filtered and the filtrate was concentrated in vacuo to afford the title compound 1d (1.73 g, 94.4% yield).

[0317] MS (ESI) m / z = 322.2 [M+H] + .

[0318] 1 H NMR (400MHz, CDCl3): δ6.65-6.50(m,3H),3.92(br d,J=10.0Hz,3H),3.70-3.62(m,1H),3.60-3.51(m,1H),3.44(br s,2H),3.32-3.00(m,3H),2.97(br s,1H),2.79(br s,1H),2.23(s,3H),1.48(s,9H).

[0319] Step 5: Tert-butyl (2S)-2-{[2-(4-bromo-2,6-difluorophenyl)-5-methyl-1H-1,3-benzodiazol-1-yl]methyl}morpholine-4-carboxylate (1e)

[0320] To a solution of compound 1d (810.0 mg, 2.52 mmol) in n-butanol (14 mL) at room temperature were added 4-bromo-2,6-difluorobenzaldehyde (612.6 mg, 2.77 mmol) and acetic acid (144 μL, 2.52 mmol). The resulting mixture was purged with nitrogen three times, and the reaction mixture was stirred at 90°C for 14 hours. LCMS indicated complete reaction of the starting material. The reaction solution was then concentrated in vacuo to afford the crude product, which was purified by flash chromatography on silica gel (ethyl acetate / petroleum ether = 0-17%) to afford the title compound 1e (189.8 mg, 14.4% yield).

[0321] MS (ESI) m / z = 524.2 [M+H] + .

[0322] 1H NMR (400MHz, CDCl3) δ7.63(br s,1H),7.37(br d,J=7.6Hz,1H),7.26(br d,J=6.0Hz,2H),7.19(br d,J=7.2Hz,1H),4.10(br s,2H),3.88-3.53(m,4H),3.30(br s,1H),2.77(br s,1H),2.51(br s,3H),2.48-2.38(m,1H),1.42(br s,9H).

[0323] Step 6: 1-[4-(1-{[(2R)-4-acetylmorpholin-2-yl]methyl}-5-methyl-1H-1,3-benzodiazol-2-yl)-3,5-difluorophenyl]pyrrolidin-2-one (1)

[0324] To a solution of compound 1e (90.0 mg, 0.172 mmol) and pyrrolidin-2-one (29.3 mg, 0.345 mmol) in 1,4-dioxane (3 mL) were added CsCO (112.3 mg, 0.345 mmol), Pd(OAc) (5.8 mg, 0.026 mmol), and Xantphos (20.0 mg, 0.034 mmol) at room temperature. The resulting mixture was then sparged with nitrogen for 5 minutes and stirred in a microwave at 100°C for 35 minutes. LCMS indicated the reaction was complete. The reaction was quenched by the addition of water (5 mL) and extracted with ethyl acetate (10 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to give a crude product, which was dissolved in dichloromethane (2 mL). Trifluoroacetic acid (400 μL) was added under an ice-water bath. The reaction mixture was stirred at room temperature for 1 hour, and LCMS indicated the reaction was complete. The mixture was concentrated under reduced pressure, and the resulting residue was dissolved in methanol (2 mL). Triethylamine (238 μL, 1.71 mmol) was added under ice-water bath to adjust the pH to 8. Acetic anhydride (80 μL, 0.854 mmol) was then added and allowed to react at room temperature for 1 hour. LCMS indicated the reaction was complete. The reaction mixture was concentrated and purified by C18 reverse-phase chromatography (acetonitrile / water (containing 0.05% ammonia) = 10-75%) and lyophilized to afford the title compound 1 (52.0 mg, 64% yield).

[0325] MS (ESI) m / z = 469.2 [M+H] + .

[0326] 1H NMR (400MHz, DMSO-d6) δ8.43(br d,J=7.0Hz,1H),7.63(dd,J=3.3,10.3Hz,2H),7.37(s,1H),6.93-6.73(m,1H),4.09(br dd,J=9.2,18.9Hz,2H),3.99-3.85(m,3H),3.77-3.52(m,2H),3.51-3.41(m,2H),3.09-2.99(m ,2H),2.90-2.82(m,1H),2.73-2.65(m,1H),2.63-2.58(m,2H),2.41(s,3H),2.18-2.03(m,2H).

[0327] Example 2

[0328] Methyl 2-((2-(2,6-difluoro-4-(2-carbonylpyrrolidin-1-yl)phenyl)-5-methyl-1H-benzo[d]imidazol-1-yl)methyl)morpholine-4-carboxylate (2)

[0329]

[0330] The synthesis steps of Example 2 refer to Example 1. In step 1, tert-butyl 2-(hydroxymethyl)morpholine-4-carboxylate is substituted for tert-butyl (2R)-2-(hydroxymethyl)morpholine-4-carboxylate, and in step 6, methyl chloroformate is substituted for acetic anhydride.

[0331] MS (ESI) m / z = 486.4 [M+H] + .

[0332] Example 3

[0333] (S)-1-(4-(1-((4-acetylmorpholin-2-yl)methyl)-5-methyl-1H-benzo[d]imidazol-2-yl)-3-chlorophenyl)pyrrolidin-2-one (3)

[0334]

[0335] The synthesis steps of Example 3 refer to Example 1, wherein in step 5, 4-bromo-2-chlorobenzaldehyde is substituted for 4-bromo-2,6-difluorobenzaldehyde.

[0336] MS (ESI) m / z = 467.2 [M+H] + .

[0337] Example 4

[0338] (S)-1-(4-(1-((4-acetylmorpholin-2-yl)methyl)-5-methyl-1H-benzo[d]imidazol-2-yl)-3-chloro-5-fluorophenyl)pyrrolidin-2-one (4)

[0339]

[0340] The synthesis steps of Example 4 refer to Example 1, wherein in step 5, 4-bromo-2-chloro-6-fluorobenzaldehyde is used instead of 4-bromo-2,6-difluorobenzaldehyde.

[0341] MS (ESI) m / z = 485.5 [M+H] + .

[0342] 1 H NMR (400MHz, DMSO-d6) δ7.88(br s,1H),7.79(br d,J=11.6Hz,1H),7.70-7.60(m,1H),7.49(br s,1H),7.20-7.12(m,1H),4.33-4.16(m,2H),4.03(br d,J=15.0Hz,1H),3.97-3.87(m,3H),3.78(br d,J=17.6Hz,1H),3.68-3.55(m,1H),3.12(br d,J=11.0Hz,1H),2.99(br d,J=11.6Hz,1H),2.81-2.70(m,2H),2.58(br d,J=7.9Hz,2H),2.44(br s,3H),2.14-2.06(m,2H),1.93(s,2H).

[0343] Example 5

[0344] (S)-1-(4-(1-((4-acetylmorpholin-2-yl)methyl)-5-(difluoromethyl)-1H-benzo[d]imidazol-2-yl)-3,5-difluorophenyl)pyrrolidin-2-one (5)

[0345]

[0346] The synthesis steps of Example 5 refer to Example 1, except that 4-(difluoromethyl)-1-fluoro-2-nitrobenzene is used in place of 1-fluoro-4-methyl-2-nitrobenzene in Step 3.

[0347] MS (ESI) m / z = 505.5 [M+H] + .

[0348] Example 6

[0349] Methyl (S)-2-((2-(2,6-difluoro-4-(methylcarbamoyl)phenyl)-6-methyl-3H-imidazo[4,5-b]pyridin-3-yl)methyl)morpholine-4-carboxylate (6)

[0350]

[0351] Step 1: Methyl (R)-2-(hydroxymethyl)morpholine-4-carboxylate (6a)

[0352] To a solution of tert-butyl (2R)-2-(hydroxymethyl)morpholine-4-carboxylate (10.00 g, 46.03 mmol) in methanol (50 mL) was added dropwise 4N HCl in methanol (57.5 mL, 230.13 mmol) at room temperature. The reaction mixture was then stirred at room temperature for 4 hours. TLC (petroleum ether / ethyl acetate = 3 / 1, sample treated with saturated NaHCO₃ solution) indicated complete reaction of the starting material, and the mixture was concentrated under reduced pressure. The resulting residue was dissolved in dichloromethane (50 mL), and ethyldiisopropylamine (38.0 mL, 230.13 mmol) and methyl chloroformate (4.3 mL, 55.23 mmol) were slowly added dropwise in an ice-water bath. The reaction mixture was then stirred at room temperature for 2 hours. TLC (petroleum ether / ethyl acetate = 1 / 2) indicated complete reaction of the starting material. The reaction was quenched by the addition of water (100 mL) and extracted with ethyl acetate (50 mL x 5). The combined organic phase was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give the title compound 6a (9.45 g, yield: 94%).

[0353] 1 H NMR (400MHz, CDCl3) δ4.02-3.85(m,3H),3.73(s,3H),3.69-3.66(m,1H),3.63-3.53(m,3H),3.05-2.92(m,1H),2.90-2.72(m,1H).

[0354] Step 2: Methyl (S)-2-((1,3-dicarbonylisoindolin-2-yl)methyl)morpholine-4-carboxylate (6b)

[0355] The synthesis steps of compound 6b refer to step 1 of Example 1, wherein compound 6a is used instead of tert-butyl (2R)-2-(hydroxymethyl)morpholine-4-carboxylate.

[0356] MS (ESI) m / z = 305.0 [M+H] + .

[0357] 1H NMR (400MHz, CDCl3) δ7.91-7.84(m,2H),7.77-7.71(m,2H),3.99(br s,1H),3.93-3.87(m,2H),3.82-3.80(m,1H),3.77(br s,1H),3.71(s,3H),3.70-3.64(m,1H),3.47(dt,J=2.4,11.2Hz,1H),3.04(br s,1H),2.83(br t,J=11.6Hz,1H).

[0358] Step 3: Methyl (S)-2-(aminomethyl)morpholine-4-carboxylate (6c)

[0359] The synthesis steps of compound 6c refer to step 2 of Example 1, wherein compound 6b is substituted for compound 1a.

[0360] 1 H NMR (400MHz, CDCl3) δ3.91(br d,J=9.2Hz,3H),3.72(s,3H),3.61-3.48(m,1H),3.37(br d,J=2.4Hz,1H),2.99(br s,1H),2.76(br d,J=6.4Hz,1H),2.74-2.63(m,1H).

[0361] Step 4: Methyl (S)-2-(((5-methyl-3-nitropyridin-2-yl)amino)methyl)morpholine-4-carboxylate (6d)

[0362] To a solution of 2-chloro-5-methyl-3-nitropyridine (200.0 mg, 1.16 mmol) and compound 6c (356.2 mg, 1.39 mmol) in dimethyl sulfoxide (2 mL) was added ethyl diisopropylamine (0.57 mL, 3.48 mmol) at room temperature, and the reaction mixture was stirred at 60°C for 12 hours. LCMS showed complete disappearance of the starting material. Ethyl acetate (30 mL) was added for dilution, the mixture was filtered, and the filtrate was concentrated under reduced pressure to afford the crude product, which was purified by flash silica gel chromatography (ethyl acetate / petroleum ether = 0-50%) to afford the title compound 6d (237.2 mg, 66% yield).

[0363] MS (ESI) m / z = 310.9 [M+H] + .

[0364] 1H NMR (400MHz, CDCl3) δ8.42-8.19(m,3H),4.09-3.83(m,3H),3.73(s,3H),3.70-3.51(m,3H),3.11-2.93(m,1H),2.89-2.69(m,1H),2.28(s,3H).

[0365] Step 5: Methyl (S)-2-(((3-amino-5-methylpyridin-2-yl)amino)methyl)morpholine-4-carboxylate (6e)

[0366] The synthesis steps of compound 6e refer to step 4 of Example 1, wherein compound 6d is substituted for compound 1c.

[0367] 1 H NMR (400MHz, CDCl3) δ7.55(s,1H),6.71(s,1H),4.31(br s,1H),4.07-3.82(m,J=12.8Hz,3H),3.72(s,3H),3.71-3.63(m,2H),3.61-3.51(m,1H),3.44-3.31(m,1H),3.21(br s,2H),3.06-2.96(m,1H),2.86-2.75(m,1H),2.16(s,3H).

[0368] Step 6: Methyl (S)-2-((2-(2,6-difluoro-4-(methylcarbamoyl)phenyl)-6-methyl-3H-imidazo[4,5-b]pyridin-3-yl)methyl)morpholine-4-carboxylate (6)

[0369] The synthesis steps of compound 6 refer to step 5 of Example 1, wherein compound 6e is substituted for compound 1d, and 3,5-difluoro-4-formyl-N-methylbenzamide is substituted for 4-bromo-2,6-difluorobenzaldehyde.

[0370] MS (ESI) m / z = 460.1 [M+H] + .

[0371] 1H NMR (400MHz, CDCl3) δ8.32(s,1H),7.92(s,1H),7.53(br s,1H),7.45(br d,J=8.8Hz,2H),4.36(dd,J=3.2,14.4Hz,1H),4.16(br dd,J=7.6,14.4Hz,1H),3.98(br s,1H),3.83-3.69(m,1H),3.66(s,3H),3.65-3.61(m,1H),3.57(br d,J=10.8Hz,1H),3.23(br t,J=10.8Hz,1H),3.03(d,J=4.4Hz,3H),2.76(br s,1H),2.53(s,3H),2.47(br s,1H).

[0372] Example 7

[0373] Methyl (S)-2-((2-(2,6-difluoro-4-(methylcarbamoyl)phenyl)-5-methyl-1H-imidazo[4,5-b]pyridin-1-yl)methyl)morpholine-4-carboxylate (7)

[0374]

[0375] Step 1: 6-Methyl-2-nitropyridin-3-yltrifluoromethanesulfonic acid (7a)

[0376] To a solution of 6-methyl-2-nitropyridine-3-ol (1.00 g, 6.50 mmol) in dichloromethane (10 mL) at 0°C under nitrogen was added triethylamine (1.4 mL, 9.70 mmol) and trifluoromethanesulfonic anhydride (1.3 mL, 7.80 mmol). The resulting solution was stirred at 0°C for 2 hours. Water (80 mL) was added to quench the reaction, and the mixture was extracted with ethyl acetate (50 mL x 3). The combined organic layers were washed with brine (80 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to afford the title compound 7a (1.70 g, yield: 92%).

[0377] 1 H NMR (400MHz, CDCl3) δ7.82 (d, J = 8.4 Hz, 1H), 7.60 (d, J = 8.4 Hz, 1H), 2.70 (s, 3H).

[0378] Step 2: Methyl (S)-2-(((6-methyl-2-nitropyridin-3-yl)amino)methyl)morpholine-4-carboxylate (7b)

[0379] To a solution of compound 7a (1.20 g, 4.2 mmol) and compound 6c (870 mg, 3.5 mmol) in acetonitrile (15 mL) was added dropwise triethylamine (1.10 g, 10.5 mmol) at room temperature. The resulting solution was stirred at 80°C for 12 hours. LCMS showed complete disappearance of the starting material. The product was concentrated under reduced pressure, and the resulting crude product was purified by reverse-phase C18 column chromatography (methanol / water (0.1% TFA) = 50%) and lyophilized to afford the title compound 7b (275.0 mg, 25% yield).

[0380] MS (ESI) m / z = 310.9 [M+H] + .

[0381] 1 H NMR (400MHz, CDCl3) δ7.92-7.80(m,1H),7.35-7.29(m,2H),3.74(s,3H),3.63-3.52(m,3H),3.5 1-3.40(m,2H),3.39-3.32(m,1H),3.25(dd,J=7.6,13.2Hz,1H),3.11-2.93(m,2H),2.52(s,3H).

[0382] Step 3: Methyl (S)-2-(((2-amino-6-methylpyridin-3-yl)amino)methyl)morpholine-4-carboxylate (7c)

[0383] The synthesis steps of compound 7c refer to step 4 of Example 1, wherein compound 7b is substituted for compound 1c.

[0384] MS (ESI) m / z = 280.9 [M+H] + .

[0385] 1 H NMR (400MHz, CDCl3) δ6.75(d,J=8.0Hz,1H),6.53(d,J=7.6Hz,1H),3.74(s,3H),3.61-3.53(m,3H),3.46( dd,J=3.6,13.6Hz,1H),3.25(dd,J=7.6,13.6Hz,1H),3.11-3.02(m,2H),2.91-2.69(m,2H),2.34(s,3H).

[0386] Step 4: Methyl (S)-2-((2-(2,6-difluoro-4-(methylcarbamoyl)phenyl)-5-methyl-1H-imidazo[4,5-b]pyridin-1-yl)methyl)morpholine-4-carboxylate (7)

[0387] The synthesis steps of compound 7 refer to step 5 of Example 1, wherein compound 7c is substituted for compound 1d, and 3,5-difluoro-4-formyl-N-methylbenzamide is substituted for 4-bromo-2,6-difluorobenzaldehyde.

[0388] MS (ESI) m / z = 460.1 [M+H] + .

[0389] 1 H NMR(400MHz, CDCl3)δ8.91(br s,1H),7.78(d,J=6.4Hz,1H),7.52(d,J=8.4Hz,2H),7.22(d,J=8.4Hz,1H),4.17-3.98(m,2H),3.95-3.59(m,6H) ,3.59-3.46(m,1H),3.36-3.18(m,1H),3.05(d,J=4.8Hz,3H),2.86-2.77(m,1H),2.75(s,3H),2.56-2.42(m,1H).

[0390] Example 8

[0391] Methyl (S)-2-((2-(2,6-difluoro-4-(methylcarbamoyl)phenyl)-6-methyl-3H-imidazo[4,5-c]pyridin-3-yl)methyl)morpholine-4-carboxylate (8)

[0392]

[0393] Step 1: (S)-5-(((4-(methoxycarbonyl)morpholin-2-yl)methyl)amino)-2-methyl-4-nitropyridine 1-oxide (8a)

[0394] 5-Bromo-2-methyl-4-nitropyridine 1-oxide (200.0 mg, 0.86 mmol) and compound 6c (329.0 mg, 1.89 mmol) were dissolved in tetrahydrofuran (5 mL) at room temperature, and the resulting solution was stirred at 80°C for 12 hours. LCMS showed complete disappearance of the starting material. The crude product was concentrated under reduced pressure, and methanol (2 mL) was added. The filtered solid was dried to afford the title compound 8a (50.0 mg, 18% yield).

[0395] MS (ESI) m / z = 327.0 [M+H] + .

[0396] 1H NMR(400MHz, CDCl3)δ8.06(s,1H),8.01(s,1H),7.90-7.80(m,1H),4.23-3.81(m,4H),3.74(s, 3H),3.61-3.55(m,1H),3.42-3.35(m,1H),3.32-3.24(m,1H),3.14-3.00(m,2H),2.41(s,3H).

[0397] Step 2: Methyl (S)-2-(((4-amino-6-methylpyridin-3-yl)amino)methyl)morpholine-4-carboxylate (8b)

[0398] The synthesis steps of compound 8b refer to step 4 of Example 1, wherein compound 8a is substituted for compound 1c.

[0399] MS (ESI) m / z = 281.2 [M+H] + .

[0400] 1 H NMR (400MHz, CDCl3) δ7.76(s,1H),6.46(s,1H),4.03-3.81(m,4H),3.73(s,3H),3.69-3.53(m,3H),3.16-3.07(m,2H),2.40(s,3H).

[0401] Step 3: Methyl (S)-2-((2-(2,6-difluoro-4-(methylcarbamoyl)phenyl)-6-methyl-3H-imidazo[4,5-c]pyridin-3-yl)methyl)morpholine-4-carboxylate (8)

[0402] The synthesis steps of compound 8 refer to step 5 of Example 1, wherein compound 8b is substituted for compound 1d, and 3,5-difluoro-4-formyl-N-methylbenzamide is substituted for 4-bromo-2,6-difluorobenzaldehyde.

[0403] MS (ESI) m / z = 460.2 [M+H] + .

[0404] 1H NMR (400MHz, CDCl3) δ8.87(s,1H),7.58(s,1H),7.49(d,J=8.4Hz,2H),6.92-6.75(m,1H),4.25-4.09(m,2H),3.98-3.71(m,3H) ,3.69(s,3H),3.65-3.55(m,1H),3.36-3.25(m,1H),3.06(d,J=4.8Hz,3H),2.90-2.77(m,1H),2.72(s,3H),2.60-2.46(m,1H).

[0405] Example 9

[0406] Methyl (S)-2-((2-(2,6-difluoro-4-(2-carbonylpyrrolidin-1-yl)phenyl)-7-methylimidazo[1,2-a]pyridin-3-yl)methyl)morpholine-4-carboxylate (9)

[0407]

[0408] Step 1: tert-Butyl (2R)-2-formylmorpholine-4-carboxylate (9a)

[0409] At -70 ° C and under nitrogen protection, dimethyl sulfoxide (9.8 mL, 138.08 mmol) was added dropwise to a solution of oxalyl chloride (9.9 mL, 115.07 mmol) in anhydrous dichloromethane (200 mL). The mixture was stirred at -70 ° C for 30 minutes. A solution of (2R)-2-(hydroxymethyl)morpholine-4-carboxylic acid tert-butyl ester (10.00 g, 46.03 mmol) in dichloromethane (30 mL) was added dropwise, and the mixture was stirred at -70 ° C for 2 hours. Triethylamine (32.0 mL, 230.13 mmol) was added dropwise at -70 ° C, and the mixture was stirred at -70 ° C for 30 minutes. TLC (petroleum ether / ethyl acetate = 1 / 1) showed that the reaction was complete. After the mixture returned to room temperature, it was washed with saturated NaHCO3 solution (80 mL × 2) and brine (80 mL). The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give the title compound 9a (9.90 g, yield: 99.9%).

[0410] 1 H NMR (400MHz, CDCl3) δ9.67(s,1H),4.15-4.04(m,1H),3.95-3.82(m,2H),3.70-3.53(m,2H),3.13-2.97(m,2H),1.50(s,9H).

[0411] Step 2: tert-Butyl (2R)-2-formylmorpholine-4-carboxylate (9b)

[0412] To a solution of compound 9a (9.90 g, 45.99 mmol) in methanol (200 mL) at 0°C under nitrogen was added potassium carbonate (25.43 g, 183.97 mmol) and dimethyl (1-diazo-2-oxopropyl)phosphonate (17.67 g, 91.99 mmol) in sequence. The mixture was stirred at room temperature for 12 hours. TLC (petroleum ether / ethyl acetate = 3 / 1) indicated completion of the reaction. Water (80 mL) was added to the mixture, and the mixture was concentrated under reduced pressure to remove the methanol. The residue was extracted with ethyl acetate (100 mL x 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash silica gel chromatography (ethyl acetate / petroleum ether = 0-25%) to afford compound 9b (6.4 g, 65.9% yield).

[0413] 1 H NMR (400MHz, CDCl3) δ4.29-4.25(m,1H),3.99-3.96(m,1H),3.93-3.63(m,1H ), 3.64-3.55 (m, 2H), 3.33-3.27 (m, 2H), 2.50 (d, J = 2.4Hz, 1H), 1.49 (s, 9H).

[0414] Step 3: Methyl (S)-2-ethynylmorpholine-4-carboxylate (9c)

[0415] To a solution of compound 9b (6.4 g, 30.29 mmol) in dichloromethane (60 mL) was added dropwise a 4N HCl solution in methanol (60 mL, 240.00 mmol) at room temperature. The reaction mixture was then stirred at room temperature for 12 hours. TLC (petroleum ether / ethyl acetate = 3 / 1) indicated complete reaction of the starting material, and the mixture was concentrated under reduced pressure. The resulting residue was dissolved in dichloromethane (80 mL), and ethyldiisopropylamine (14.8 mL, 89.43 mmol) and methyl chloroformate (2.8 mL, 35.98 mmol) were slowly added dropwise under an ice-water bath. The reaction mixture was then stirred at room temperature for 12 hours. TLC (petroleum ether / ethyl acetate = 1 / 2) indicated complete reaction of the starting material, and the reaction solution was concentrated under reduced pressure. The resulting residue was purified by flash silica gel chromatography (ethyl acetate / petroleum ether = 0-25%) to afford the title compound 9c (5.00 g, 97.6% yield).

[0416] 1H NMR (400MHz, CDCl3) δ4.31-4.28(m,1H),4.01-3.97(m,1H),3.90-3.82(m,1H ), 3.75 (s, 3H), 3.68-3.56 (m, 2H), 3.74-3.34 (m, 2H), 2.52 (d, J = 2.0Hz, 1H).

[0417] Step 4: Methyl (S)-2-((2-(4-bromo-2,6-difluorophenyl)-7-methylimidazo[1,2-a]pyridin-3-yl)methyl)morpholine-4-carboxylate (9d)

[0418] Compound 9c (7.5 g, 44.33 mmol), 4-bromo-2,6-difluorobenzaldehyde (9.80 g, 44.33 mmol), and 4-methylpyridin-2-amine (4.79 g, 44.33 mmol) were dissolved in toluene (75 mL). Under nitrogen, cuprous chloride (1.32 g, 13.30 mmol) and copper trifluoromethanesulfonate (4.81 g, 13.30 mmol) were added. The mixture was stirred at 85°C for 5 minutes, and then N,N-dimethylacetamide (1.2 mL, 13.30 mmol) was added. After the reaction mixture was stirred at 85°C for 12 hours, TLC indicated the complete disappearance of the starting material. Water (50 mL) and aqueous ammonia (30 mL) were added. The reaction mixture was extracted with dichloromethane (100 mL × 3), the combined organic phases were dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (ethyl acetate / petroleum ether = 0-85%) to give compound 9d (6.00 g, yield: 26.8%).

[0419] MS (ESI) m / z = 480.1 [M+H] + .

[0420] 1 H NMR (400MHz, CDCl3) δ8.17(d,J=6.8Hz,1H),7.38(s,1H),7.25-7.18(m,2H),6.67(br d,J=6.8Hz,1H),4.01-3.74(m,3H),3.68(s,3H),3.56(br s,1H),3.44-3.34(m,1H),3.06-2.85(m,3H),2.60(dd,J=11.2,12.8Hz,1H),2.42(s,3H).

[0421] Step 5: Methyl (S)-2-((2-(2,6-difluoro-4-(2-carbonylpyrrolidin-1-yl)phenyl)-7-methylimidazo[1,2-a]pyridin-3-yl)methyl)morpholine-4-carboxylate (9)

[0422] To a solution of compound 9d (48.0 mg, 0.10 mmol) in 1,4-dioxane (2 mL) in a microwave oven, 2-pyrrolidone (34.0 mg, 0.40 mmol), N,N'-dimethylethylenediamine (6 μL, 0.05 mmol), cesium carbonate (195.5 mg, 0.60 mmol), and cuprous iodide (9.5 mg, 0.05 mmol) were added sequentially. The reaction mixture was heated to 150°C under microwave conditions for 1 hour. LCMS indicated the reaction was complete. The reaction mixture was concentrated under reduced pressure, and the residue was dissolved in acetonitrile and filtered. The filtrate was directly purified by C18 reverse-phase chromatography (acetonitrile / water (containing 0.05% NH3·H2O) = 5-95%, flow rate: 60 mL / min) and lyophilized to afford the title compound 9 (26.0 mg, 53.7% yield).

[0423] MS (ESI) m / z = 485.3 [M+H] + .

[0424] 1 H NMR(400MHz,DMSO-d6)δ8.40(d,J=7.3Hz,1H),7.60(d,J=10.0Hz,2H),7.34(s,1H),6.8 2(dd,J=1.4,7.2Hz,1H),3.88(t,J=7.2Hz,2H),3.75-3.60(m,3H),3.55(s,3H),3.46(br d,J=7.3Hz,1H),3.28-3.19(m,1H),3.01(br d,J=6.0Hz,2H),2.80(br s,1H),2.57(t,J=8.0Hz,3H),2.37(s,3H),2.08(quin,J=7.6Hz,2H).

[0425] Example 10

[0426] (S)-1-(4-(1-((4-acetylmorpholin-2-yl)methyl)-5-chloro-1H-benzo[d]imidazol-2-yl)-3,5-difluorophenyl)pyrrolidin-2-one (10)

[0427]

[0428] Step 1: (R)-2-((4-acetylmorpholin-2-yl)methyl)isoindoline-1,3-dione (10a)

[0429] Compound 1a (5.00 g, 14.44 mmol) was dissolved in 4M HCl in 1,4-dioxane (20 mL), and the mixture was stirred at 25°C for 2 hours. TLC (petroleum ether / ethyl acetate = 1 / 1) indicated the reaction was complete. After concentration, the residue was dissolved in tetrahydrofuran (30 mL), and triethylamine (9.8 mL, 70.74 mmol) and acetic anhydride (4.0 mL, 42.44 mmol) were added under an ice-water bath. The mixture was allowed to react at room temperature for 1.5 hours. TLC (petroleum ether / ethyl acetate = 1 / 1) indicated the reaction was complete. The reaction was quenched by the addition of water (50 mL) and extracted with ethyl acetate (50 mL x 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash silica gel chromatography (ethyl acetate / petroleum ether = 0-100%) to afford compound 10a (2.50 g, 54.0% yield).

[0430] MS (ESI) m / z = 314.1 [M+H] + .

[0431] 1 H NMR (400MHz, CDCl3) δ7.88 (br s, 2H), 7.75 (br d, J = 5.4Hz, 2H), 4.55-4.28 (m, 1H), 3.94 (br d,J=7.2Hz,2H),3.83-3.64(m,3H),3.60-3.42(m,1H),3.38-3.02(m,1H),2.93-2.61(m,1H),2.10(br d,J=6.0Hz,3H).

[0432] Step 2: (S)-1-(2-(aminomethyl)morpholino)ethan-1-one (10b)

[0433] To a solution of compound 10a (1.00 g, 3.47 mmol) in ethanol (6 mL) was added hydrazine hydrate (206 μL, 4.16 mmol) at room temperature. The reaction mixture was then stirred at 80°C for 1 hour. TLC (petroleum ether / ethyl acetate = 1 / 1) indicated the reaction was complete. The mixture was cooled to room temperature, filtered, and the filtrate was concentrated under reduced pressure. The residue was dissolved in ethyl acetate (20 mL) and filtered. The filtrate was concentrated under reduced pressure to give the title compound 10b (600.0 mg, 98.4% yield).

[0434] 1H NMR (400MHz, CDCl3) δ4.42-4.32(m,1H),3.95-3.87(m,1H),3.68-3.43(m,3H),3.42-3.18 (m,2H),3.03-2.91(m,1H),2.88-2.57(m,3H),2.46(dd,J=10.8,13.2Hz,1H),2.07(s,3H).

[0435] Step 3: (S)-1-(2-(((4-chloro-2-nitrophenyl)amino)methyl)morpholino)ethan-1-one (10c)

[0436] To a solution of compound 10b (300.0 mg, 1.90 mmol) in tetrahydrofuran (5 mL) was added 1-fluoro-4-chloro-2-nitrobenzene (399.5 mg, 2.28 mmol) and potassium carbonate (523.0 mg, 3.79 mmol) at room temperature, and the reaction mixture was stirred at 25°C for 2 hours. LCMS showed complete disappearance of the starting material. Water (10 mL) was added, and the mixture was extracted with ethyl acetate (15 mL x 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to afford the crude product, which was purified by flash silica gel chromatography (ethyl acetate / petroleum ether = 0-50%, 35 mL / min) to afford the title compound 10c (300.0 mg, 30.2% yield).

[0437] MS (ESI) m / z = 314.1 [M+H] + .

[0438] 1 H NMR (400MHz, CDCl3) δ8.27-8.15(m,2H),7.46-7.37(m,1H),6.89-6.79(m,1H),4.54(br d,J=13.2Hz,1H),4.02(dd,J=2.8,10.8Hz,1H),3.77-3.53(m,3H),3.51-3.28(m,3H),2.67(dd,J=10.8,13.2Hz,1H),2.15-2.10(m,3H).

[0439] Step 4: (S)-1-(2-(((2-amino-4-chlorophenyl)amino)methyl)morpholino)ethan-1-one (10d)

[0440] To a solution of compound 10c (400.0 mg, 1.28 mmol) in tetrahydrofuran (5 mL), ethanol (5 mL), and water (5 mL) at room temperature were added ammonium chloride (675.7 mg, 12.75 mmol) and iron powder (356.0 mg, 6.38 mmol). The reaction mixture was then stirred at 60°C for 2 hours. LCMS showed complete disappearance of the starting material. The mixture was filtered and the filtrate was extracted with ethyl acetate (30 mL x 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to afford the title compound 10d (380.0 mg, 94.5% yield).

[0441] 1 H NMR (400MHz, CDCl3) δ6.78-6.67(m,2H),6.57-6.51(m,1H),4.56-4.38(m,1H),4.03-3.93(m,1H), 3.71-3.50(m,5H),3.36-3.25(m,1H),3.23-3.07(m,3H),2.87-2.60(m,1H),2.11(d,J=3.2Hz,3H).

[0442] Step 5: (S)-1-(2-((2-(4-bromo-2,6-difluorophenyl)-5-chloro-1H-benzo[d]imidazol-1-yl)methyl)morpholino)ethan-1-one (10e)

[0443] To a solution of compound 10d (190.0 mg, 0.67 mmol) in n-butanol (5 mL) was added 4-bromo-2,6-difluorobenzaldehyde (163.0 mg, 0.74 mmol) and acetic acid (238 μL, 1.34 mmol) at room temperature. The resulting mixture was stirred at 90°C for 12 hours. LCMS indicated complete reaction. The reaction mixture was then concentrated in vacuo to afford the crude product, which was purified by flash chromatography on silica gel (petroleum ether / ethyl acetate = 0-80%) to afford the title compound 10e (300.0 mg, 83.1% yield).

[0444] 1 H NMR (400MHz, CDCl3) δ7.86-7.79(m,1H),7.46-7.40(m,1H),7.38-7.27(m,3H) ,4.50-4.31(m,1H),4.16-4.10(m,3H),3.85-3.70(m,1H),3.55-3.47(m,1H), 3.35-3.25(m,1H),3.18-3.06(m,1H),2.37-2.26(m,1H),2.05-2.05(m,3H).

[0445] Step 6: (S)-1-(4-(1-((4-acetylmorpholin-2-yl)methyl)-5-chloro-1H-benzo[d]imidazol-2-yl)-3,5-difluorophenyl)pyrrolidin-2-one (10)

[0446] To a solution of compound 10e (50.0 mg, 0.172 mmol) and pyrrolidin-2-one (8.7 mg, 0.10 mmol) in 1,4-dioxane (3 mL) was added CsCO (67.2 mg, 0.21 mmol), Pd(dba) (5.9 mg, 0.01 mmol), and Xantphos (11.9 mg, 0.02 mmol) at room temperature. The resulting mixture was then sparged with nitrogen for 5 minutes and stirred at 90°C for 12 hours. LCMS indicated the reaction was complete. The reaction solution was cooled to room temperature, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by preparative HPLC (column: YMC-Actus Triart C18 150*30 mm*7 μm, acetonitrile / water (containing 0.05% ammonia) = 47-70%, 9 minutes) and lyophilized to afford the title compound 10 (13.5 mg, 26.8% yield).

[0447] MS (ESI) m / z = 489.1 [M+H] + .

[0448] 1 H NMR (400MHz, DMSO-d6) δ7.82-7.73(m,2H),7.66(d,J=10.8Hz,2H),7.36(dd,J=2.0,8.8Hz,1H),4.33(br dd,J=3.2,15.2Hz,1H),4.17(br dd,J=7.6,15.2Hz,1H),3.92(t,J=7.2Hz,2H),3.69-3.48(m,3H),3.21(br d,J=10.0Hz,2H),3.10(br s,1H),3.00(br s, 1H), 2.60 (t, J = 8.0Hz, 2H), 2.17-2.11 (m, 2H), 1.93 (s, 3H).

[0449] Example 11

[0450] (S)-1-(4-(1-((4-acetylmorpholin-2-yl)methyl)-6-fluoro-5-methyl-1H-benzo[d]imidazol-2-yl)-3-chloro-5-fluorophenyl)pyrrolidin-2-one (11)

[0451]

[0452] Step 1: (S)-1-(2-(((5-fluoro-4-methyl-2-nitrophenyl)amino)methyl)morpholino)ethan-1-one (11a)

[0453] The synthesis steps of compound 11a refer to step 3 of Example 10, wherein 1,5-difluoro-2-methyl-4-nitrobenzene is used instead of 1-fluoro-4-chloro-2-nitrobenzene.

[0454] MS (ESI) m / z = 312.1 [M+H] + .

[0455] 1 H NMR(400MHz, CDCl3)δ8.24(br s,1H),8.11-8.04(m,1H),6.53-6.45(m,1H),4.54(br d,J=13.3Hz,1H),4.07-4.01(m,1H),3.74-3.58(m,3H),3.46-3.26 (m,3H),2.66(dd,J=10.8,13.1Hz,1H),2.20-2.12(m,6H).

[0456] Step 2: (S)-1-(2-((2-(4-bromo-2-chloro-6-fluorophenyl)-6-fluoro-5-methyl-1H-benzo[d]imidazol-1-yl)methyl)morpholino)ethan-1-one (11b)

[0457] To a mixed solution of compound 11a (100.0 mg, 0.32 mmol) in ethanol (2 mL) and DMSO (2 mL) at room temperature were added 4-bromo-2-chloro-6-fluorobenzaldehyde (83.9 mg, 0.35 mmol) and Na2S2O4 (447.4 mg, 2.57 mmol). The resulting mixture was stirred at 90°C for 12 hours. LCMS indicated complete reaction of the starting material. The reaction was quenched by the addition of water (15 mL) and extracted with ethyl acetate (20 mL x 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by C18 reverse-phase chromatography (acetonitrile / water (containing 0.05% NH3·H2O) = 10-75%, flow rate: 40 mL / min) and lyophilized to afford the title compound 11b (100.0 mg, yield: 62.5%).

[0458] MS (ESI) m / z = 498.1 [M+H] + .

[0459] Step 3: (S)-1-(4-(1-((4-acetylmorpholin-2-yl)methyl)-6-fluoro-5-methyl-1H-benzo[d]imidazol-2-yl)-3-chloro-5-fluorophenyl)pyrrolidin-2-one (11)

[0460] The synthesis steps of compound 11 refer to step 6 of Example 10, wherein compound 11b is substituted for compound 10e.

[0461] MS (ESI) m / z = 503.4 [M+H] + .

[0462] Example 12

[0463] (S)-1-(4-(1-((4-acetylmorpholin-2-yl)methyl)-5-chloro-6-fluoro-1H-benzo[d]imidazol-2-yl)-3-chloro-5-fluorophenyl)pyrrolidin-2-one (12)

[0464]

[0465] Step 1: tert-Butyl (S)-2-(((4-chloro-5-fluoro-2-nitrophenyl)amino)methyl)morpholine-4-carboxylate (12a)

[0466] The synthesis steps of compound 12a refer to step 3 of Example 10, wherein 1-chloro-2,4-difluoro-5-nitrobenzene is used instead of 1-fluoro-4-chloro-2-nitrobenzene.

[0467] 1 H NMR (400MHz, CDCl3) δ8.36-8.25 (m, 2H), 6.65 (d, J = 11.6Hz, 1H), 4.09-3.80 (m,3H),3.75-3.67(m,1H),3.63-3.53(m,1H),3.43-3.35(m,1H),3.34-3.25 (m,1H),2.99(brs,1H),2.79(br s,1H),1.48(s,9H).

[0468] Step 2: tert-Butyl (S)-2-(((2-amino-4-chloro-5-fluorophenyl)amino)methyl)morpholine-4-carboxylate (12b)

[0469] The synthesis steps of compound 12b refer to step 4 of Example 10, wherein compound 12a is substituted for compound 10c.

[0470] 1H NMR (400MHz, DMSO-d6) δ7.81-7.65(m,1H),6.63-6.37(m,1H),4.68(br s,1H),4.40-4.18(m,1H),3.84(br d,J=12.0Hz,1H),3.76-3.61(m,2H),3.12-3.00(m,1H),2.85(br s,1H),2.53(br s,2H),1.40(d,J=1.6Hz,9H),1.27-1.20(m,3H).

[0471] Step 3: Tert-butyl (S)-2-((2-(4-bromo-2-chloro-6-fluorophenyl)-5-chloro-6-fluoro-1H-benzo[d]imidazol-1-yl)methyl)morpholine-4-carboxylate (12c)

[0472] The synthesis steps of compound 12c refer to step 5 of Example 10, wherein compound 12b is substituted for compound 10d, and 4-bromo-2-chloro-6-fluorobenzaldehyde is substituted for 4-bromo-2,6-difluorobenzaldehyde.

[0473] MS (ESI) m / z = 578.1 [M+H] + .

[0474] 1 H NMR (400MHz, CDCl3) δ7.87(d,J=6.8Hz,1H),7.59-7.55(m,1H),7.40-7.33(m,2H),4.08-3.97(m,2H),3.75(br s,3H),3.62-3.50(m,1H),3.33(br s,1H),2.81(br s,1H),2.47(br t,J=11.6Hz,1H),0.96(s,9H).

[0475] Step 4: (S)-1-(2-((2-(4-bromo-2-chloro-6-fluorophenyl)-5-chloro-6-fluoro-1H-benzo[d]imidazol-1-yl)methyl)morpholino)ethan-1-one (12d)

[0476] The synthesis steps of compound 12c refer to step 1 of Example 10, wherein compound 12c is substituted for compound 1a.

[0477] MS (ESI) m / z = 518.0 [M+H] + .

[0478] 1H NMR (400MHz, CDCl3) δ7.90-7.85(m,1H),7.61-7.56(m,1H),7.41-7.32(m,2H),4.50-4.38(m,1H),4.08-4.01(m,2H),3.86-3.79(m,1H),3.55(br d,J=11.2Hz,2H),3.41-3.32(m,1H),3.24-3.11(m,1H),2.42-2.27(m,1H),2.09(s,3H).

[0479] Step 5: (S)-1-(4-(1-((4-acetylmorpholin-2-yl)methyl)-5-chloro-6-fluoro-1H-benzo[d]imidazol-2-yl)-3-chloro-5-fluorophenyl)pyrrolidin-2-one (12)

[0480] The synthesis steps of compound 12 refer to step 6 of Example 10, wherein compound 12d is used instead of compound 10e.

[0481] MS (ESI) m / z = 523.3 [M+H] + .

[0482] 1 H NMR (400MHz, DMSO-d6) δ7.93-7.87(m,3H),7.78(dd,J=2.0,12.0Hz,1H),4.38-3.99(m,3H),3.93(t,J=7.2Hz,2H),3. 72-3.44(m,3H),3.33-3.13(m,2H),3.06-2.85(m,1H),2.60(t,J=8.4Hz,2H),2.13(quin,J=7.6Hz,2H),1.95(s,3H).

[0483] Example 13

[0484] (S)-1-(4-(1-((4-acetylmorpholin-2-yl)methyl)-5-chloro-7-fluoro-1H-benzo[d]imidazol-2-yl)-3,5-difluorophenyl)pyrrolidin-2-one (13)

[0485]

[0486] Step 1: (S)-1-(2-(((4-chloro-2-fluoro-6-nitrophenyl)amino)methyl)morpholino)ethan-1-one (13a)

[0487] The synthesis steps of compound 13a refer to step 3 of Example 10, wherein 5-chloro-1,2-difluoro-3-nitrobenzene is used instead of 1-fluoro-4-chloro-2-nitrobenzene.

[0488] MS (ESI) m / z = 332.2 [M+H] + .

[0489] 1 H NMR (400MHz, CDCl3) δ8.05-7.92(m,2H),7.27-7.20(m,1H),4.54-4.40(m,1H),4.05-3.97(m,1H),3.83 -3.73(m,1H),3.71-3.60(m,2H),3.60-3.51(m,2H),3.36-3.06(m,1H),2.87-2.55(m,1H),2.12(s,3H).

[0490] Step 2: (S)-1-(2-(((2-amino-4-chloro-6-fluorophenyl)amino)methyl)morpholino)ethan-1-one (13b)

[0491] The synthesis steps of compound 13b refer to step 4 of Example 10, wherein compound 13a is substituted for compound 10c.

[0492] 1 H NMR (400MHz, CDCl3) δ6.53-6.45(m,2H),4.42(br d,J=13.2Hz,1H),4.13(q,J=7.2Hz,3H),4.00(br d,J=11.6Hz,1H),3.63-3.44(m,3H),3.36-3.25(m,1H),3.09-2.92(m,2H),2.85-2.53(m,1H),2.09(d,J=9.2Hz,3H).

[0493] Step 3: (S)-1-(2-((2-(4-bromo-2,6-difluorophenyl)-5-chloro-7-fluoro-1H-benzo[d]imidazol-1-yl)methyl)morpholino)ethan-1-one (13c)

[0494] The synthesis steps of compound 13c refer to step 5 of Example 10, wherein compound 13b is substituted for compound 10d.

[0495] MS (ESI) m / z = 503.8 [M+H] + .

[0496] 1H NMR (400MHz, CDCl3) δ7.67-7.60(m,1H),7.33-7.27(m,2H),7.14-7.05(m,1H),4.51(br d,J=13.2Hz,1H),4.41-4.27(m,2H),3.73-3.58(m,2H),3.28(br t,J=11.6Hz,1H),3.15-3.03(m,1H),2.81(br t,J=12.2Hz,1H),2.64-2.53(m,1H),2.34-2.25(m,1H),2.05(s,3H).

[0497] Step 4: (S)-1-(4-(1-((4-acetylmorpholin-2-yl)methyl)-5-chloro-7-fluoro-1H-benzo[d]imidazol-2-yl)-3,5-difluorophenyl)pyrrolidin-2-one (13)

[0498] The synthesis steps of compound 13 refer to step 6 of Example 10, wherein compound 13c is substituted for compound 10e.

[0499] MS (ESI) m / z = 507.2 [M+H] + .

[0500] 1 H NMR (400MHz, DMSO-d6) δ7.79-7.65(m,3H),7.41(d,J=11.2Hz,1H),4.51-4.37(m,1H),4.23(br d,J=13.2Hz,1H),4.11(br dd,J=8.4,15.2Hz,1H),4.02(br d,J=13.2Hz,1H),3.90(t,J=7.2Hz,2H),3.73(br d,J=12.8Hz,1H),3.63-3.50(m,2H),3.28-3.07(m,1H),3.02-2.72(m,1H) ,2.59(t,J=8.0Hz,2H),2.30-2.20(m,1H),2.57-2.07(m,2H),1.93(s,3H).

[0501] Example 14

[0502] (S)-1-(4-(1-((4-acetylmorpholin-2-yl)methyl)-5-(methyl-d3)-1H-benzo[d]imidazol-2-yl)-3,5-difluorophenyl)pyrrolidin-2-one (14)

[0503]

[0504] Step 1: tert-Butyl (S)-2-(((4-bromo-2-nitrophenyl)amino)methyl)morpholine-4-carboxylate (14a)

[0505] To a solution of compound 1b (2.00 g, 7.40 mmol) in 1,4-dioxane (10 mL) was added 4-bromo-1-fluoro-2-nitrobenzene (1.63 g, 7.40 mmol) and triethylamine (2.0 mL, 14.80 mmol) at room temperature. The reaction mixture was then stirred at 80°C for 12 hours. LCMS analysis showed complete disappearance of the starting material. The reaction mixture was then concentrated under reduced pressure to afford the crude product, which was then purified by flash silica gel chromatography (ethyl acetate / petroleum ether = 0-15%, 60 mL / min) to afford the title compound 14a (2.00 g, 58.4% yield).

[0506] 1 H NMR (400MHz, CDCl3) δ8.25 (d, J = 2.4Hz, 1H), 8.15 (br s,1H),7.44(dd,J=2.4,9.2Hz,1H),6.71(d,J=9.2Hz,1H),3.96-3.72(m,3H),3. 63(tdd,J=3.2,7.2,10.4Hz,1H),3.55-3.46(m,1H),3.39-3.23(m,2H),2.92(br s,1H),2.71(br s,1H),1.40(s,9H).

[0507] Step 2: tert-Butyl (S)-2-(((4-(methyl-d3)-2-nitrophenyl)amino)methyl)morpholine-4-carboxylate (14b)

[0508] To a mixed solution of compound 14a (1.10 g, 2.64 mmol) and 4,4,5,5-tetramethyl-2-(methyl-d3)-1,3,2-dioxaborolane (1.15 g, 7.93 mmol) in 1,4-dioxane (15 mL) and water (5 mL) was added Cs2CO3 (2.58 g, 7.93 mmol) and Pd(dppf)Cl2.CH2Cl2 (58.8 mg, 0.072 mmol) at room temperature. The resulting mixture was then sparged with nitrogen for 5 minutes and stirred at 100°C under a nitrogen atmosphere for 12 hours. LCMS indicated the reaction was complete. The reaction solution was cooled to room temperature, water (20 mL) was added, and extraction was performed with ethyl acetate (50 mL x 3). The combined organic phase was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a crude product which was purified by flash silica gel chromatography (ethyl acetate / petroleum ether = 0-20%, 35 mL / min) to give the title compound 14b (470.0 mg, yield: 45.2%).

[0509] MS (ESI) m / z = 299.2 [M+H-56] + .

[0510] 1 H NMR (400MHz, DMSO-d6) δ8.09(br t,J=5.6Hz,1H),7.87(d,J=2.4Hz,1H),7.39(dd,J=2.0,8.8Hz,1H),7.04(d,J=8.8Hz,1H),3.87(br d,J=11.2Hz,2H),3.70(br d,J=13.2Hz,1H),3.63-3.50(m,2H),3.47-3.36(m,2H),3.27-3.24(m,1H),2.88(br s,1H),1.39(s,9H).

[0511] Step 3: Tert-butyl (S)-2-((2-(4-bromo-2,6-difluorophenyl)-5-(methyl-d3)-1H-benzo[d]imidazol-1-yl)methyl)morpholine-4-carboxylate (14c)

[0512] To a solution of compound 14b (200.0 mg, 0.56 mmol) in ethanol (6 mL) and water (2 mL) at room temperature were added 4-bromo-2,6-difluorobenzaldehyde (124.7 mg, 0.56 mmol) and Na2S2O4 (294.8 mg, 1.69 mmol). The resulting mixture was stirred at 80°C under a nitrogen atmosphere for 4 hours. LCMS indicated complete reaction of the starting material. The reaction mixture was filtered, and water (3 mL) was added to the filtrate, which was then extracted with ethyl acetate (10 mL x 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to afford the title compound 14c (340.0 mg, 114.7% yield).

[0513] MS (ESI) m / z = 525.1 [M+H] + .

[0514] Step 4: (S)-1-(4-(1-((4-acetylmorpholin-2-yl)methyl)-5-(methyl-d3)-1H-benzo[d]imidazol-2-yl)-3,5-difluorophenyl)pyrrolidin-2-one (14)

[0515] To a solution of compound 14c (300.0 mg, 0.57 mmol) and pyrrolidin-2-one (48.4 mg, 0.57 mmol) in 1,4-dioxane (8 mL) were added CsCO (372.1 mg, 1.14 mmol), Pd(dba) (32.8 mg, 0.057 mmol), and Xantphos (66.1 mg, 0.11 mmol) at room temperature. The resulting mixture was then sparged with nitrogen for 5 minutes and stirred at 100°C for 2 hours. LCMS indicated the reaction was complete. After the reaction solution cooled to room temperature, p-toluenesulfonic acid (981.5 mg, 5.70 mmol) was added, and the reaction mixture was stirred at 100°C for 1 hour. LCMS indicated the reaction was complete. The reaction mixture was cooled to room temperature and methanol (5 mL) was added. Triethylamine (1.2 mL, 8.55 mmol) was added under an ice-water bath to adjust the pH to 8. Acetic anhydride (267 μL, 2.85 mmol) was then added and allowed to react at room temperature for 1 hour. LCMS indicated the reaction was complete. Water (10 mL) was added and the mixture was extracted with ethyl acetate (20 mL x 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC (column: YMC Triart C18 250*50 mm*7 μm, acetonitrile / water (containing 0.05% ammonia) = 12-52%, 9 minutes) and lyophilized to afford the title compound 14 (90.2 mg, 33.3% yield).

[0516] MS (ESI) m / z = 472.2 [M+H] + .

[0517] 1 H NMR (400MHz, DMSO-d6) δ7.69-7.58(m,3H),7.49(d,J=1.2Hz,1H),7.16(dd,J=1.6,8.4Hz,1H),4.27(br dd,J=3.6,15.2Hz,1H),4.12(br dd,J=7.2,15.2 Hz,1H),3.92(t,J=7.2Hz,2H),3.64(br dd,J=2.0,11.6Hz,1H),3.54(br s,2H),3.23(br s,2H),3.14-3.09(m,2H),2.60(t,J=8.0Hz,2H),2.18-2.08(m,2H),1.93(s,3H).

[0518] Example 15

[0519] (S)-1-(3-chloro-4-(1-((4-(cyclopropylcarbonyl)morpholin-2-yl)methyl)-5-methyl-1H-benzo[d]imidazol-2-yl)-5-fluorophenyl)pyrrolidin-2-one (15)

[0520]

[0521] Step 1: Tert-butyl (S)-2-((2-(4-bromo-2-chloro-6-fluorophenyl)-5-methyl-1H-benzo[d]imidazol-1-yl)methyl)morpholine-4-carboxylate (15a)

[0522] The synthesis steps of compound 15a refer to step 5 of Example 1, wherein 4-bromo-2-chloro-6-fluorobenzaldehyde is used instead of 4-bromo-2,6-difluorobenzaldehyde.

[0523] MS (ESI) m / z = 540.1 [M+H+2] + .

[0524] Step 2: (R)-1-(3-chloro-5-fluoro-4-(5-methyl-1-(morpholin-2-ylmethyl)-1H-benzo[d]imidazol-2-yl)phenyl)pyrrolidin-2-one (15b)

[0525] To a solution of compound 15a (870.0 mg, 1.62 mmol) and pyrrolidin-2-one (549.7 mg, 6.46 mmol) in 1,4-dioxane (10 mL) were added CsCO (1.05 g, 3.23 mmol), Pd(dba) (92.9 mg, 0.16 mmol), and Xantphos (186.8 mg, 0.32 mmol) at room temperature. The resulting mixture was then sparged with nitrogen for 5 minutes and stirred at 100°C for 4 hours. LCMS indicated the reaction was complete. After the reaction solution cooled to room temperature, p-toluenesulfonic acid monohydrate (3.07 g, 16.14 mmol) was added. The reaction mixture was stirred at 100°C for 1 hour, which indicated completion by LCMS. Water (15 mL) was added, and the mixture was extracted with ethyl acetate (20 mL x 3). The aqueous phase was adjusted to pH 8 with 1N NaOH aqueous solution and extracted with ethyl acetate (20 mL×3). The combined organic phases were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give the title compound 15b (680.0 mg, yield: 95.1%).

[0526] MS (ESI) m / z = 443.5 [M+H] + .

[0527] Step 3: (S)-1-(3-chloro-4-(1-((4-(cyclopropylcarbonyl)morpholin-2-yl)methyl)-5-methyl-1H-benzo[d]imidazol-2-yl)-5-fluorophenyl)pyrrolidin-2-one (15)

[0528] To a solution of compound 15b (50.0 mg, 0.11 mmol) in DMF (1 mL) were added cyclopropylcarboxylic acid (19.4 mg, 0.23 mmol), HATU (85.8 mg, 0.0.23 mmol), and N,N-diisopropylethylamine (75 μL, 0.45 mmol) in sequence. The mixture was stirred at room temperature for 1 hour. The reaction solution was then directly purified by C18 reverse-phase chromatography (acetonitrile / water (containing 0.05% ammonia) = 10-75%) and lyophilized to afford the title compound 15 (32.0 mg, 55.5% yield).

[0529] MS (ESI) m / z = 469.4 [M+H] + .

[0530] 1 H NMR (400MHz, DMSO-d6, t=75℃) δ7.87(s,1H),7.77(dd,J=12.0,2.1Hz,1H),7.60(d,J=8.3Hz,1H),7.49(s,1H),7.16(d d,J=8.4,1.6Hz,1H),4.27-4.11(m,2H),4.08(t,J=5.1Hz,2H),3.99(d,J=13.5Hz,1H),3.93(t,J=7.1Hz,2H),3.68(br s,1H),3.56(br s,1H),3.47(dd,J=7.0,5.1Hz,2H),3.25(t,J=11.7Hz,1H),2.59(dd,J=8.5,7.6Hz,2H),2.46(s,3H),2.13(p,J=7.5Hz,2H),1.08(t,J=7.0Hz,4H).

[0531] Example 16

[0532] (S)-1-(3-chloro-5-fluoro-4-(5-methyl-1-((4-(3,3,3-trifluoropropionyl)morpholin-2-yl)methyl)-1H-benzo[d]imidazol-2-yl)phenyl)pyrrolidin-2-one (16)

[0533]

[0534] The synthesis steps of compound 16 refer to step 3 of Example 15, wherein 3,3,3-trifluoropropionic acid is used instead of cyclopropylcarboxylic acid.

[0535] 1 H NMR (400MHz, DMSO-d6, t=75℃) δ7.86 (s, 1H), 7.76 (dd, J=12.0, 2.1Hz, 1H), 7.60 (d,J=8.3Hz,1H),7.49(s,1H),7.16(dd,J=8.5,1.6Hz,1H),4.41-4.13(m,2H),4 .08(t,J=5.1Hz,2H),3.93(t,J=7.1Hz,2H),3.78-3.64(m,1H),3.59-3.44(m,5H ), 3.35-3.16 (m, 1H), 2.59 (t, J = 8.1Hz, 2H), 2.46 (s, 3H), 2.13 (p, J = 7.6Hz, 2H).

[0536] Example 17

[0537] (S)-6-Chloro-8-fluoro-7-(5-methyl-1-((4-propionylmorpholin-2-yl)methyl)-1H-benzo[d]imidazol-2-yl)-2H-benzo[b][1,4]oxazin-3(4H)-one (17)

[0538]

[0539] Step 1: 4-Chloro-2-fluoro-6-nitrophenol (17a)

[0540] To a solution of 4-chloro-2-fluorophenol (10.00 g, 68.24 mmol) in acetic acid (100 mL) was slowly added dropwise fuming nitric acid (4.51 g, 71.65 mmol) under ice-water bath. The reaction mixture was then stirred under ice-water bath for 1 hour. LCMS showed complete disappearance of the starting material. While stirring, the reaction solution was slowly poured into water (1.5 L). After stirring for 1 hour, the solid was filtered and dried to give the title compound 17a (11.00 g, 84% yield).

[0541] 1 H NMR (400MHz, CDCl3) δ10.38 (s, 1H), 7.94 (t, J = 2.4Hz, 1H), 7.45 (dd, J = 2.4, 9.6Hz, 1H).

[0542] Step 2: 2-Amino-4-chloro-6-fluorophenol (17b)

[0543] To a solution of compound 17a (5.00 g, 26.10 mmol) in tetrahydrofuran (60 mL), ethanol (60 mL), and water (30 mL) at room temperature were added ammonium chloride (13.96 g, 261.04 mmol) and iron powder (11.69 g, 208.83 mmol). The reaction mixture was then stirred at 60°C for 2 hours. LCMS showed complete disappearance of the starting material. The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure. The residue was dispersed with ethyl acetate (500 mL) and filtered. The filtrate was then concentrated under reduced pressure to afford the title compound 17b (4.20 g, 99.6% yield).

[0544] MS (ESI) m / z = 162.0 [M+H] + .

[0545] 1 H NMR (400MHz, CDCl3) δ6.56-6.49(m,2H),5.03(br s,1H),3.93(br s,2H).

[0546] Step 3: 6-Chloro-8-fluoro-2H-benzo[b][1,4]oxazin-3(4H)-one (17c)

[0547] To a solution of compound 17b (4.20 g, 26.00 mmol) in tetrahydrofuran (50 mL) was added potassium carbonate (10.78 g, 77.99 mmol) and chloroacetyl chloride (3.1 mL, 38.99 mmol) under an ice-water bath. The mixture was then stirred at 40°C for 15 hours. LCMS showed complete disappearance of the starting material. The reaction was quenched by the addition of water (100 mL) and extracted with dichloromethane (100 mL x 3). The organic phase was dried and concentrated, and the crude product was purified by silica gel column chromatography (ethyl acetate / petroleum ether = 30%, flow rate: 30 mL / min) to afford the title compound 17c (4.30 g, 82.0%).

[0548] 1 H NMR (400MHz, DMSO-d6) δ11.00 (br s, 1H), 7.10 (dd, J = 2.4, 10.4Hz, 1H), 6.74 (t, J = 2.0Hz, 1H), 4.68 (s, 2H).

[0549] Step 4: 6-Chloro-8-fluoro-3-carbonyl-3,4-dihydro-2H-benzo[b][1,4]oxazine-7-carbaldehyde (17d)

[0550] To a solution of 2,2,6,6-tetramethylpiperidine (3.8 mL, 22.32 mmol) in anhydrous tetrahydrofuran (4 mL) at -78°C was added a 2.5 M solution of n-butyllithium in n-hexane (8.9 mL, 22.32 mmol) dropwise. After the addition was complete, the mixture was stirred at -78°C for 20 minutes, followed by the slow dropwise addition of a solution of compound 17c (1.50 g, 7.44 mmol) in anhydrous tetrahydrofuran (15 mL). After stirring at -78°C for 3 hours, N,N-dimethylformamide (1.09 g, 14.88 mmol) was slowly added dropwise. The mixture was then slowly warmed to room temperature and the reaction continued for 30 minutes. After TLC (petroleum ether / ethyl acetate = 1 / 1) indicated the reaction was complete, the mixture was quenched with saturated aqueous ammonium chloride and extracted with ethyl acetate (50 mL x 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated to yield the title compound 17d (600.0 mg, 35.0%).

[0551] 1 H NMR (400MHz, DMSO-d6) δ11.38 (br s, 1H), 10.16 (d, J = 0.8Hz, 1H), 6.85 (d, J = 1.6Hz, 1H), 4.76 (s, 2H).

[0552] Step 5: Tert-butyl (S)-2-((2-(6-chloro-8-fluoro-3-carbonyl-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)-5-methyl-1H-benzo[d]imidazol-1-yl)methyl)morpholine-4-carboxylate (17e)

[0553] The synthesis steps of compound 17e refer to step 3 of Example 14, wherein compound 1c is substituted for compound 14b, and compound 17d is substituted for 4-bromo-2,6-difluorobenzaldehyde.

[0554] MS (ESI) m / z = 531.2 [M+H] + .

[0555] 1 H NMR (400MHz, CDCl3) δ9.81 (br s,1H),7.69(s,1H),7.41(dd,J=2.4,8.4Hz,1H),7.21(d,J=8.4Hz,1H),6.79(dd,J=1.2,5.6Hz,1H),4.75-4.71(m,2H),4.12-4.02 (m,2H),3.99-3.69(m,3H),3.66-3.56(m,1H),3.41-3.26(m,1H),2.90-2.76(m,1H),2.52(s,3H),2.52-2.44(m,1H),1.43(s,9H).

[0556] Step 6: (S)-6-Chloro-8-fluoro-7-(5-methyl-1-((4-propionylmorpholin-2-yl)methyl)-1H-benzo[d]imidazol-2-yl)-2H-benzo[b][1,4]oxazin-3(4H)-one (17)

[0557] To a solution of compound 17e (182.0 mg, 0.34 mmol) in dichloromethane (5 mL) was slowly added dropwise a 4M HCl solution in 1,4-dioxane (0.9 mL) under an ice-water bath. The mixture was stirred at room temperature for 4 hours. TLC (petroleum ether / ethyl acetate = 1 / 1) indicated the reaction was complete. After concentration, the residue was dissolved in dichloromethane (3 mL). Triethylamine (240 μL, 1.72 mmol) and propionyl chloride (33 μL, 0.38 mmol) were added under an ice-water bath and allowed to react at room temperature for 2 hours. TLC (petroleum ether / ethyl acetate = 1 / 1) indicated the reaction was complete. The reaction was quenched by the addition of water (20 mL) and extracted with dichloromethane (20 mL x 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash silica gel chromatography (ethyl acetate / petroleum ether = 0-100%) to afford compound 17 (74.0 mg, 44.7% yield).

[0558] MS (ESI) m / z = 487.1 [M+H] + .

[0559] Biological tests

[0560] Test Example 1: In vitro biological activity evaluation

[0561] The FLIPR assay was used to screen the antagonistic activity of compounds on hP2X3 and hP2X2 / 3 receptors (the effect of the compound on the ion channel was indicated by the change of calcium flux signal).

[0562] 1. Experimental instruments and materials

[0563]

[0564]

[0565] 2. Experimental steps

[0566] 1321N1 cells stably transfected with hP2X3 and hP2X2 / 3 receptor (adherent cells) were digested, centrifuged, resuspended in plating medium (DMEM + 10% DFBS) and counted. The cells were adjusted to 2*10 5Cells / mL were plated in 384-well assay plates, 50 μL of cells were plated per well, and cultured in a 5% CO2, 37°C incubator for 16-24 hours. Test compounds were prepared with DMSO at 180 times the required concentration (20 mM DMSO stock solution), 500 nL was taken per well and added to the 384-well compound plate, supplemented with 30 μL of FLIPR buffer (containing 1.26 mM CaCl2). 2+ Prepare 3 times the required concentration of agonist α,β-meATP in FLIPR buffer (500nM for hP2X3 cells and 1000nM for hP2X2 / 3 cells) and add 35μL of agonist to each well of another 384-well compound plate. Remove the plate of cells that have been cultured for 16-24 hours, aspirate the cell supernatant, and add 30μL of dye ( Calcium 4 detection kit, diluted with FLIPR buffer) and incubated for 1 hour. 15 μL of compound was added to each well of cells (FLIPR instrument loading). After 15 minutes, 22.5 μL of agonist was added to each well and the fluorescence signal was detected (excitation wavelength 470nm-495nm, emission wavelength 515nm-575nm). The difference between the peak and valley values ​​of the signal was taken as the basic data. The highest concentration data of the positive drug was taken as 100% inhibition rate, and the DMSO data was taken as 0% inhibition rate. The inhibitory effect curve of the compound was fitted on the software Graphpad Prism 6 and the IC was calculated. 50 value.

[0567] Table 1. Half-maximal inhibitory concentration (IC50) of the compounds of the present disclosure on hP2X3 and hP2X2 / 3 receptors 50 )

[0568] Compound number <![CDATA[hP2X3(IC 50 ,nM)]]> <![CDATA[hP2X2 / 3(IC 50 ,nM)]]> MK-7264 35.4 116.2

[0569] 1 36.8 5566 2 105.0 18990 3 41.2 1805 4 31.6 1927 5 100.4 NT 6 >10000 NT 7 5843 NT 8 >10000 NT 9 75.8 30270 10 54.1 7407 11 79.4 NT 12 87.7 NT 13 765.9 NT 14 33.0 NT 15 85.1 NT 16 70.5 NT 17 45.3 3501

[0570] NT: Not Tested.

[0571] Test Example 2: CYP inhibition experiment

[0572] The metabolic reactions of representative substrates of the five major human CYP isoforms (CYP1A2, CYP2C9, CYP2C19, CYP2D6, and CYP3A4 / 5) were evaluated using 150 pooled human liver microsomes (Corning, Cat. No. 452117). The effects of varying concentrations of the test compounds on the metabolic reactions of phenacetin (CYP1A2), diclofenac sodium (CYP2C9), S-mephenytoin (CYP2C19), bufuralol hydrochloride (CYP2D6), and midazolam (CYP3A4 / 5) were determined by liquid chromatography-tandem mass spectrometry (LC / MS / MS).

[0573] A 200 μL reaction mixture (100 mmol / L phosphate buffer, pH 7.4, containing 0.3% DMSO, 0.6% acetonitrile, and 0.1% methanol, by volume) containing 30 μM phenacetin, 10 μM diclofenac sodium, 35 μM S-mephenytoin, 5 μM bufuralol hydrochloride, 3 μM midazolam, 1 mM NADPH, test compounds (at concentrations of 0.1, 0.3, 1, 3, 10, and 30 μmol / L), positive compounds, or a blank control, was incubated with mixed human liver microsomes (0.2 mg / mL) for 5 minutes at 37°C. Then, 200 μL of an acetonitrile solution containing 3% formic acid and 40 nM internal standard verapamil was added, and the mixture was centrifuged at 4000 rpm for 50 minutes. The mixture was cooled on ice for 20 minutes and then centrifuged at 4000 rpm for 20 minutes to precipitate the protein. A 200 μL supernatant was used for LC / MS / MS analysis.

[0574] The peak area was calculated based on the chromatogram. The residual activity ratio (%) was calculated using the following formula:

[0575] Peak area ratio = metabolite peak area / internal standard peak area

[0576] Residual activity ratio (%) = peak area ratio of the test compound group / peak area ratio of the blank group

[0577] CYP half-maximal inhibitory concentration (IC 50 ) was calculated using Excel XLfit 5.3.1.3.

[0578] The measured CYP half-maximal inhibition concentration (IC50) values ​​are shown in the table below.

[0579] Table 2. Half maximal inhibitory concentration (IC50) of some compounds of the present disclosure on CYP 50 )

[0580]

[0581] Test Example 3: In vitro metabolic stability test of human hepatocytes

[0582] The compound concentrations in the reaction system were determined by LC / MS / MS to calculate the intrinsic clearance of the test compounds and to evaluate their in vitro metabolic stability in human hepatocytes.

[0583] 247.5 μL of 1×10 6 The reaction was initiated by adding a mixture of 100 μM human hepatocytes (purchased from Bioreclamation IVT, Catalog No. S01205) and 2.5 μL of 100 μM test compound or positive control to the incubation plate. The reaction was incubated at 37°C and 600 rpm. At 0.5, 5, 15, 30, 45, 60, 80, 100, and 120 minutes, 20 μL of the incubation system was transferred to the stop plate. Vortex to mix for 2 minutes. The stop plate was centrifuged at 4000 rpm for 20 minutes. 40 μL of the supernatant of each compound was transferred to a 96-well sample plate, and 160 μL of purified water was added to dilute the sample.

[0584] The resulting samples were quantified by ion chromatogram. The residual rate was calculated based on the peak area of ​​the test compound or positive control. The slope k was determined by linear regression of the natural logarithm of the residual rate against the incubation time using Microsoft Excel.

[0585] Intrinsic clearance (in vitro CL int , μL / min / 10 6 cells) was calculated from the slope value according to the following equation:

[0586] in vitro CL int =kV / N

[0587] V = incubation volume (0.25 mL);

[0588] N = number of cells per well (0.25 × 10 6 cell)

[0589] The measured human hepatocyte intrinsic clearance values ​​are shown in Table 3.

[0590] Table 3. Intrinsic clearance of some compounds in this disclosure by human hepatocytes

[0591]

[0592]

[0593] Test Example 4: Caco-2 permeability test

[0594] The apparent permeability coefficient (P) of the drug was determined by liquid chromatography tandem mass spectrometry (LC / MS / MS) using a Caco-2 cell model. app ).

[0595] The transport rate of the drug from the top to the base was measured. 5 108 μL of HBSS (25 mM HEPES, pH 7.4) containing 10 μM of the test compound was added to the top of a Transwell (from Corning) chamber containing Caco-2 cells (from ATCC) at a density of 5 cells / cm2. At the same time, 8 μL of the sample was immediately taken out and added to a new 96-well plate containing 72 μL of HBSS (25 mM HEPES, pH 7.4) and 240 μL of acetonitrile solution (containing 100 nM alprazolam, 200 nM caffeine and 100 nM tolbutamide) as the initial drug-addition end sample (AB). The plate was vortexed at 1000 rpm for 10 minutes, and 300 μL of HBSS (25 mM HEPES, pH 7.4) was added to the base.

[0596] To measure the drug transport rate from the basolateral to the apical side, add 308 μL of HBSS (25 mM HEPES, pH 7.4) containing 10 μM of the test compound to the basolateral wells. Immediately remove 8 μL of the sample and transfer it to a new 96-well plate containing 72 μL of HBSS (25 mM HEPES, pH 7.4) and 240 μL of acetonitrile solution (containing 100 nM alprazolam, 200 nM caffeine, and 100 nM tolbutamide) as the initial drug-addition sample (BA). Vortex at 1000 rpm for 10 minutes, and then add 100 μL of HBSS (25 mM HEPES, pH 7.4) to the apical side.

[0597] Both tip-to-base and base-to-tip testing are performed simultaneously.

[0598] After the transport cycle, 8 μL of sample was taken from the dosing port (apical for AB flow and basolateral for BA flow) and added to 72 μL of HBSS (25 mM HEPES, pH 7.4) and 240 μL of acetonitrile (containing 100 nM alprazolam, 200 nM caffeine, and 100 nM tolbutamide) in a new 96-well plate. 80 μL of sample was taken directly from the basolateral for AB flow and the apical for BA flow, and added to 240 μL of acetonitrile (containing 100 nM alprazolam, 200 nM caffeine, and 100 nM tolbutamide) in a new 96-well plate. The sample was vortexed at 1000 rpm for 10 minutes. The sample was centrifuged at 4000 rpm for 30 minutes. 100 μL of supernatant was transferred to a new 96-well plate. All samples were mixed with 100 μL of purified water for LC-MS / MS analysis.

[0599] Data were calculated using Microsoft Excel, and peak areas were calculated from the chromatograms. The apparent permeability coefficient (Papp) is in cm / s and is calculated using the following formula:

[0600]

[0601] Papp is the apparent permeability (cm / s×10 -6 );

[0602] dQ / dt is the drug delivery rate (pmol / s);

[0603] A is the surface area of ​​the membrane (cm 2 );

[0604] D0 is the initial drug concentration at the supply end (nM; pmol / cm3).

[0605] The outflow ratio can be determined by the following formula:

[0606]

[0607] Papp(BA) is the apparent permeability coefficient from the base to the apex;

[0608] Papp(AB) is the apparent permeability from the apical end to the basal end.

[0609] The measured apparent permeability coefficient values ​​of Caco-2 cells are shown in Table 4.

[0610] Table 4

[0611]

[0612] Note: BLU-5937.

[0613] Test Example 5: Pharmacokinetics experiment in rats

[0614] The LC / MS / MS method was used to determine the drug concentration in the plasma of rats at different times after oral administration of the disclosed compound, to study the pharmacokinetic behavior of the disclosed compound in rats and to evaluate its pharmacokinetic characteristics.

[0615] 1. Experimental plan

[0616] 1.1 Investigational Drugs

[0617] Compound 4 and BLU-5937.

[0618] 1.2 Experimental animals

[0619] 6-8 week old, SPF grade, male, healthy SD rats, 3 rats per group.

[0620] 1.3 Drug preparation

[0621] Oral administration: Weigh a certain amount of drug and add 0.5% hydropropyl methylcellulose, 0.1% Tween 80 and 99.4% water to prepare a 1 mg / mL white suspension.

[0622] 1.4 Administration

[0623] SD rats were fasted overnight and then gavaged with compound 4 and BLU-5937 at a dose of 5 mg / kg.

[0624] 2. Operation

[0625] Rats were orally administered with the disclosed compounds. 0.2 mL of blood was collected from the jugular vein at 0.25, 0.5, 1, 2, 4, 8, and 24 hours after administration. The blood was placed in a tube containing EDTA-K2 and centrifuged at 4°C, 4000 rpm for 5 minutes to separate the plasma, which was then stored at -75°C.

[0626] Determination of the test compound content in rat plasma after oral administration of different drug concentrations: 50 μL of rat plasma at each time point after administration was taken, 200 μL of acetonitrile solution of internal standard dexamethasone (50 ng / mL) was added, vortexed for 30 seconds, and centrifuged at 4°C and 4700 rpm for 15 minutes. The supernatant of the plasma sample was diluted three times with water, and 2.0 μL was taken for LC / MS / MS analysis.

[0627] 3. Pharmacokinetic parameter results

[0628] The pharmacokinetic parameters of some compounds disclosed in this disclosure in rats are as follows:

[0629] Table 5

[0630]

Claims

1. A compound of formula (I) or a pharmaceutically acceptable salt thereof, which is a compound of formula (I-1) or a pharmaceutically acceptable salt thereof in, R1 is selected from hydrogen, deuterium, halogen, C1-C6 alkyl optionally substituted by deuterium; R2 are each independently selected from hydrogen; R5 is selected from C1-C3 alkyl; R 6a and R 6b are each independently selected from hydrogen, deuterium, chlorine atoms and fluorine atoms; R7 is R8 is selected from hydrogen; Alternatively, R7 and R8 together with the atoms to which they are attached form a heterocyclic ring A m is an integer from 1 to 3.

2. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to claim 1, which is 3. A method for preparing the compound of formula (I-1) or a pharmaceutically acceptable salt thereof according to claim 1, comprising the following steps: The compound represented by formula (Ia) reacts with the compound represented by formula (Ib)-1 under alkaline conditions to obtain the compound represented by formula (Ic)-1; the compound represented by formula (Ic)-1 is subjected to a reduction reaction to obtain the compound represented by formula (Id)-1; the compound represented by formula (Id)-1 and the compound represented by formula (Ie)-1 are subjected to a ring-closure reaction under acidic conditions to obtain the compound represented by formula (I-1); R1, R2, R5, R 6a 、R 6b , R7, R8 and m are as defined in claim 1; and Y is halogen.

4. A method for preparing the compound of formula (I-1) or a pharmaceutically acceptable salt thereof according to claim 1, comprising the following steps: The compound represented by formula (Ia) reacts with the compound represented by formula (Ib)-1 under alkaline conditions to obtain the compound represented by formula (Ic)-1; the compound represented by formula (Ic)-1 is subjected to a reduction reaction to obtain the compound represented by formula (Id)-1; the compound represented by formula (Id)-1 and the compound represented by formula (If)-1 are subjected to a ring-closure reaction under acidic conditions to obtain the compound represented by formula (Ig)-1; the compound represented by formula (Ig)-1 is reacted in the presence of a catalyst to obtain the compound represented by formula (I-1); The catalyst is selected from palladium / carbon, Raney nickel, tetrakistriphenylphosphine palladium, palladium dichloride, palladium acetate, [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium, 1,1'-bis(dibenzylphosphino)dichlorodipentaferropalladium, tris(dibenzylideneacetone)dipalladium or 2-dicyclohexylphosphine-2',6'-dimethoxybiphenyl, [1,1''-bis(di-tert-butylphosphino)ferrocene]dichloropalladium(II), cuprous iodide, cuprous bromide, cuprous chloride, and copper trifluoromethanesulfonate; R1, R2, R5, R 6a 、R 6b , R7, R8 and m are as defined in claim 1; and Y and Z are each independently selected from halogen.

5. A pharmaceutical composition comprising the compound according to claim 1 or 2 or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable excipient.

6. Use of the compound according to claim 1 or 2 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 5, in the preparation of a medicament for treating a disease associated with P2X3 activity.

7. The method according to claim 6, wherein the disease is selected from the group consisting of pain, urinary tract disease and cough.

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