MAS-related g protein-coupled receptor x2 antagonist and use thereof

Abstract

Provided in the present invention is a compound as shown in formula I-A, or a tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug thereof. The compound has a relatively good antagonistic effect against MAS-related G protein-coupled receptor X2.

Description

MAS-related G protein receptor X2 antagonist and use thereof

[0001] This application requires the applicant to:

[0002] Priority benefit of the prior application, patent application number 202410071703.7, filed with the State Intellectual Property Office of China on January 17, 2024, entitled “MAS-related G protein receptor X2 antagonists and uses thereof”;

[0003] Priority benefit of the prior application, patent application number 202410166244.0, filed with the State Intellectual Property Office of China on February 5, 2024, entitled “MAS-related G protein receptor X2 antagonists and uses thereof”;

[0004] Priority benefit of the prior application, patent application number 202410843343.8, filed with the State Intellectual Property Office of China on June 26, 2024, entitled “MAS-related G protein receptor X2 antagonists and uses thereof”;

[0005] Priority benefit of the prior application, patent application number 202411283294.3, filed with the State Intellectual Property Office of China on September 12, 2024, entitled “MAS-related G protein receptor X2 antagonists and uses thereof”;

[0006] The entire contents of said prior application are incorporated into the present application by reference. Technical Field

[0007] The present invention belongs to the field of medicine, and in particular, relates to a MAS-related G protein receptor X2 antagonist and its use. Background Art

[0008] Mast cells (MCs) are tissue-resident immune cells that originate from the hematopoietic lineage. MCs reside within various connective tissues and vascularized organs. They are found in greatest numbers and density at the interface between the internal and external milieu, responding to foreign organisms and antigens and acting as sentinels. These sites include the dermis, skeletal muscle, oral mucosa, gastrointestinal mucosa and submucosa, conjunctiva, alveoli and airways, and auricles. Dermal MCs are often in close proximity to blood vessels, nerves, and lymphatic vessels. Activation of MCs through the human high-affinity immunoglobulin E (IgE) receptor (FcεRI) and Mas-related G protein-coupled receptor X2 (MRGPRX2) leads to degranulation, the release of various bioactive substances (such as histamine and proteases), and the de novo synthesis of prostaglandins, leukotrienes, and several cytokines, playing a central role in host defense, inflammation, and allergic responses.

[0009] Mas-related G protein-coupled receptors (MRGPRs) are divided into nine subfamilies based on the receptor members: MRGPRA, B, C, D, E, F, G, H, and the primate-specific MRGPRX. Each subfamily includes different subtypes, such as MRGPRX1, MRGPRX2, MRGPRX3, and MRGPRX4. Transcriptome analysis shows that most MRGPR family members are expressed in peripheral neurons, but MRGPRX2 is primarily expressed in skin MCs, with an expression rate even higher than that of Fcε-RI. MRGPRX2 can sense a variety of endogenous or exogenous agonists, including polycationic compounds and peptides, to trigger mast cell degranulation.

[0010] Studies have shown that inappropriate activation of MRGPRX2 may lead to mast cell-related diseases such as drug pseudoallergy, rosacea, atopic dermatitis, allergic contact dermatitis, urticaria, pruritus, mastocytosis, interstitial cystitis, pain, rheumatoid arthritis, asthma, and ulcerative colitis. Currently, the IND application for the MRGPRX2 antagonist EP262 has been approved by the FDA for the treatment of mast cell-related diseases (such as chronic urticaria). Preclinical studies have shown that EP262 can effectively inhibit mast cell activation and degranulation induced by multiple MRGPRX2 agonists and block the release of trypsin and inflammatory factors in human mast cells. Oral administration of EP262 can effectively inhibit agonist-induced mast cell degranulation and increased vascular permeability in MRGPRX2-KI mice, showing significant potential for the treatment of mast cell-related diseases. All of this indicates that the development of new MRGPRX2 antagonists for the treatment of various mast cell-related diseases is a promising direction.

[0011] Currently, there are no drugs on the market that act as MRGPRX2 antagonists. Therefore, the development of new compounds that can antagonize the activity of MAS-related G protein receptor X2 has positive significance for the treatment of diseases. Summary of the Invention

[0012] The purpose of the present invention is to provide a new compound as a MAS-related G protein receptor X2 antagonist.

[0013] In a first aspect of the present invention, there is provided a compound of formula IA, a tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug thereof:

[0014] in,

[0015] X 1 、X 2 、X 3 、X 4 、X 5 and X 6Each independently represents a ring atom;

[0016] X 1 、X 2 、X 3 、X 5 and X 6 are each independently N, CH2, CH or C;

[0017] X 4 is C;

[0018] X 1 With X 2 Between, X 5 With X 6 The bond between them is a single bond or a double bond;

[0019] X 5 and X 6 The linked group fragment In the middle, A and X 5 、X 6 The ring atoms together form a 6-10 membered aryl, a 3-11 membered heterocycloalkyl or a 5-10 membered heteroaryl; the heteroatoms are independently selected from one or more of N, O and S; and the A is further replaced by R c Replaced by; said R c Substitution is one or more substitutions, when the substituent R c When there are multiple substituents, the substituents are the same or different;

[0020] Ring B is -(CH2) 0-2 -C 3-12 Cycloalkyl, -(CH2) 0-2 -3-10 membered heterocycloalkyl or -(CH2) 0-2 -5-10 membered heteroaryl;

[0021] Ring C is a 6- to 10-membered aryl group or a 5- to 10-membered heteroaryl group;

[0022] R a and R c Each is independently H, halogen, hydroxy, amino, cyano, carbonyl, oxo, C 1-6 Alkyl, C 3-8 Cycloalkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Halogenated alkyl, C 3-8 Halogenated cycloalkyl, C 1-6 Alkyl hydroxyl, C 1-6 Alkylcarbonyl, C 1-6 Alkoxy, C 1-6 Halogenated alkoxy or 3-10 membered heterocycloalkyl; the C 1-6 Alkyl, C3-8 Cycloalkyl, C 2-6 Alkynyl, C 1-6 Halogenated alkyl, C 3-8 Halogenated cycloalkyl, C 1-6 Alkyl hydroxyl, C 1-6 Alkylcarbonyl, C 1-6 Alkoxy and C 1-6 The haloalkoxy group is optionally substituted with one or more R d Substituted; said R d A substituent selected from the group consisting of halogen, hydroxy, amino, nitro, cyano, carbonyl, oxo, carboxyl, C 1-6 Alkyl, C 2-6 Alkynyl, C 1-6 Halogenated alkyl, C 1-6 Alkyl hydroxyl, C 1- 6-alkylcarbonyl, C 1-6 Alkoxy, C 1-6 Haloalkoxy; when the substituent R d When there are multiple R d Same or different;

[0023] R b Halogen 、 Hydroxyl, amino, cyano, carbonyl, oxo, C 1-6 Alkyl, C 1-6 Alkylamino, C 3-8 Cycloalkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Halogenated alkyl, C 3-8 Halogenated cycloalkyl, C 1-6 Alkyl hydroxyl, C 1-6 Alkylcarbonyl, C 1-6 Alkoxy, C 1-6 Haloalkoxy, 3-10 membered heterocycloalkyl, -(CHR j ) 0-3 -S(=O)(=NR j )-R k 、-(CHR j ) 1-3 -S(O)2-R k 、-(CHR j ) 1-3 -OC(O)-N(R k )2、-(CHR j ) 0-3 -(N=)S(=O)(R k )2 or -(CHR j ) 0-3 -P(=O)(R k )2; said C 1-6 Alkyl, C1-6 Alkylamino, C 3-8 Cycloalkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Halogenated alkyl, C 3- 8-halogenated cycloalkyl, C 1-6 Alkyl hydroxyl, C 1-6 Alkylcarbonyl, C 1-6 Alkoxy, C 1-6 Haloalkoxy, 3-10 membered heterocycloalkyl, -(CHR j ) 0-3 -S(=O)(=NR j )-R k 、-(CHR j ) 1-3 -S(O)2-R k 、-(CHR j ) 1-3 -OC(O)-N(R k )2、-(CHR j ) 0-3 -(N=)S(=O)(R k )2 and -(CHR j ) 0-3 -P(=O)(R k )2 is optionally replaced by one or more R d Substituted; said R d A substituent selected from the group consisting of halogen, hydroxy, amino, nitro, cyano, carbonyl, oxo, carboxyl, C 1-6 Alkyl, C 2-6 Alkynyl, C 1-6 Halogenated alkyl, C 1-6 Alkyl hydroxyl, C 1-6 Alkylcarbonyl, C 1-6 Alkoxy, C 1-6 Haloalkoxy; when the substituent R d When there are multiple R d Same or different;

[0024] n is 1, 2, 3 or 4, and R b At least one of -(CHR j ) 0-3 -S(=O)(=NR j )-R k 、-(CHR j ) 1-3 -S(O)2-R k 、-(CHR j ) 1-3 -OC(O)-N(R k )2、-(CHR j) 0-3 -(N=)S(=O)(R k )2 or -(CHR j ) 0-3 -P(=O)(R k )2;

[0025] R 1 、R 2 and R 3 are independently H, halogen, cyano, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Halogenated alkyl, C 3-8 Cycloalkyl, halogenated C 3-8 Cycloalkyl, -OR f 、-C(O)OR f 、-OC(O)R f 、-N(R f )2、-N(R f )C(O)R f 、

[0026] -N(R f )S(O)2R f or -S(O)2R f ; the R f Selected from H, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Alkylamino,

[0027] -(CH2)rR g , 6-10 membered aromatic group, C 3-8 Cycloalkyl, 5-10 membered heteroaryl or 5-10 membered heterocycloalkyl, or two R f The group and the atoms to which it is attached together form a 5- to 11-membered heterocycloalkyl group; the R g H, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-8 Cycloalkyl, 6- to 10-membered aryl, 5- to 10-membered heteroaryl, or 5- to 10-membered heterocycloalkyl;

[0028] R j and R k are independently H, cyano, amino, C 1-6 Alkyl, C 1-6 Alkylamino, -(CH2) 0-3 C 3-8 Cycloalkyl, -COC 1-6 Alkyl, C 1-6 haloalkyl, C substituted by hydroxy 1-6 Alkyl, a single 5- to 10-membered heterocycloalkyl, or when the R k When there are two, they form a 3-10 membered heterocyclic alkyl group with the connected N, S or P atom; the amino, C 1-6 Alkylamino, -(CH2) 0-3 C 3-8 Cycloalkyl, -COC 1-6 Alkyl, C 1-6 Haloalkyl, C substituted by hydroxy 1-6 Alkyl, a single 5- to 10-membered heterocycloalkyl, and R k The 3-10 membered heterocycloalkyl formed with the attached N, S or P atom is optionally substituted by one or more R m Substituted; said R m A substituent selected from the group consisting of halogen, hydroxy, amino, nitro, cyano, carbonyl, oxo, carboxyl, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Alkyl hydroxyl, C 1-6 Alkylcarbonyl, C 1-6 Alkoxy, C 1-6 Halogenated alkoxy, C 3-8 Cycloalkyl; when the substituent R m When there are multiple R m Same or different;

[0029] m and r are 0, 1, 2, or 3 respectively.

[0030] In a second aspect of the present invention, there is provided a compound of formula I, a tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug thereof:

[0031] in,

[0032] X 1 、X 2 、X 3 、X 4 、X 5 and X 6 Each independently represents a ring atom;

[0033] X 1 、X 2 、X 3 、X 5 and X 6 are each independently N, CH2, CH or C;

[0034] X 4 is C;

[0035] X1 With X 2 Between, X 5 With X 6 The bond between them is a single bond or a double bond;

[0036] X 5 and X 6 The linked group fragment In the middle, A and X 5 、X 6 The ring atoms together form a 6-10 membered aryl, a 3-11 membered heterocycloalkyl or a 5-10 membered heteroaryl; the heteroatoms are independently selected from one or more of N, O and S; and the A is further replaced by R c Replaced by; said R c Substitution is one or more substitutions, when the substituent R c When there are multiple substituents, the substituents are the same or different;

[0037] Ring B is -(CH2) 0-2 -C 3-12 Cycloalkyl, -(CH2) 0-2 -3-10 membered heterocycloalkyl or -(CH2) 0-2 -5-10 membered heteroaryl;

[0038] Ring C is a 6- to 10-membered aryl group or a 5- to 10-membered heteroaryl group;

[0039] R a and R c Each is independently H, halogen, hydroxy, amino, cyano, carbonyl, oxo, C 1-6 Alkyl, C 3-8 Cycloalkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Halogenated alkyl, C 3-8 Halogenated cycloalkyl, C 1-6 Alkyl hydroxyl, C 1-6 Alkylcarbonyl, C 1-6 Alkoxy, C 1-6 Halogenated alkoxy or 3-10 membered heterocycloalkyl; the C 1-6 Alkyl, C 3-8 Cycloalkyl, C 2-6 Alkynyl, C 1-6 Halogenated alkyl, C 3-8 Halogenated cycloalkyl, C 1-6 Alkyl hydroxyl, C 1-6 Alkylcarbonyl, C 1-6 Alkoxy and C 1-6 The haloalkoxy group is optionally substituted with one or more R d Substituted; said R dA substituent selected from the group consisting of halogen, hydroxy, amino, nitro, cyano, carbonyl, oxo, carboxyl, C 1-6 Alkyl, C 2-6 Alkynyl, C 1-6 Halogenated alkyl, C 1-6 Alkyl hydroxyl, C 1- 6-alkylcarbonyl, C 1-6 Alkoxy, C 1-6 Haloalkoxy; when the substituent R d When there are multiple R d Same or different;

[0040] R b Halogen 、 Hydroxyl, amino, cyano, carbonyl, oxo, C 1-6 Alkyl, C 3-8 Cycloalkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Halogenated alkyl, C 3-8 Halogenated cycloalkyl, C 1-6 Alkyl hydroxyl, C 1-6 Alkylcarbonyl, C 1-6 Alkoxy, C 1-6 Haloalkoxy, 3-10 membered heterocycloalkyl, -(CHR j ) 0-3 -S(=O)(=NR j )-R k 、-(CHR j ) 1-3 -S(O)2-R k 、-(CHR j ) 1-3 -OC(O)-N(R k )2、-(CHR j ) 0-3 -(N=)S(=O)(R k )2 or -(CHR j ) 0-3 -P(=O)(R k )2; said C 1-6 Alkyl, C 3-8 Cycloalkyl, C 2-6 Alkynyl, C 1-6 Halogenated alkyl, C 3-8 Halogenated cycloalkyl, C 1-6 Alkyl hydroxyl, C 1-6 Alkylcarbonyl, C 1-6 Alkoxy and C 1-6 The haloalkoxy group is optionally substituted with one or more R d Substituted; said R dA substituent selected from the group consisting of halogen, hydroxy, amino, nitro, cyano, carbonyl, oxo, carboxyl, C 1-6 Alkyl, C 2-6 Alkynyl, C 1-6 Halogenated alkyl, C 1-6 Alkyl hydroxyl, C 1-6 Alkylcarbonyl, C 1-6 Alkoxy, C 1-6 Haloalkoxy; when the substituent R d When there are multiple R d Same or different;

[0041] n is 1, 2, 3 or 4, and R b At least one of -(CHR j ) 0-3 -S(=O)(=NR j )-R k 、-(CHR j ) 1-3 -S(O)2-R k 、-(CHR j ) 1-3 -OC(O)-N(R k )2、-(CHR j ) 0-3 -(N=)S(=O)(R k )2 or -(CHR j ) 0-3 -P(=O)(R k )2;

[0042] R 1 、R 2 and R 3 are independently H, halogen, cyano, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Halogenated alkyl, C 3-8 Cycloalkyl, halogenated C 3-8 Cycloalkyl, -OR f 、-C(O)OR f 、-OC(O)R f 、-N(R f )2、-N(R f )C(O)R f 、

[0043] -N(R f )S(O)2R f or -S(O)2R f ; the R f Selected from H, C 1-6 Alkyl, C2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Alkylamino,

[0044] -(CH2)rR g , 6-10 membered aromatic group, C 3-8 Cycloalkyl, 5-10 membered heteroaryl or 5-10 membered heterocycloalkyl, or two R f The group and the atoms to which it is attached together form a 5- to 11-membered heterocycloalkyl group; the R g H, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-8 Cycloalkyl, 6- to 10-membered aryl, 5- to 10-membered heteroaryl, or 5- to 10-membered heterocycloalkyl;

[0045] R j and R k are independently H, cyano, amino, C 1-6 Alkyl, C 3-8 Cycloalkyl, C 1-6 Haloalkyl, single 5-10 membered heterocycloalkyl, or when the R k When there are two, they form a 5-10 membered heterocyclic alkyl group with the connected N, S or P atom; the amino, C 1-6 Alkyl, C 3-8 Cycloalkyl, C 1-6 haloalkyl, a single 5- to 10-membered heterocycloalkyl, and R k The 5- to 10-membered heterocycloalkyl formed with the attached N, S or P atom is optionally replaced by one or more R m Substituted; said R m A substituent selected from the group consisting of halogen, hydroxy, amino, nitro, cyano, carbonyl, oxo, carboxyl, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Alkyl hydroxyl, C 1-6 Alkylcarbonyl, C 1-6 Alkoxy, C 1-6 Halogenated alkoxy, C 3-8 Cycloalkyl; when the substituent R m When there are multiple R m Same or different;

[0046] m, and r are 0, 1, 2 or 3 respectively.

[0047] In the present invention, the definitions of certain substituents in the compounds of Formula IA or Formula I may be as described below, and the definitions of substituents not mentioned are as described in any of the above schemes.

[0048] In a preferred embodiment of the present invention, the compound shown in Formula IA is selected from the following structures:

[0049] Among them, A and X 5 、X 6 The ring atoms together form a 6-8 membered aryl group, a 3-8 membered heterocycloalkyl group, a 5-membered heteroaromatic ring or a 6-membered heteroaromatic ring; the heteroatoms are independently selected from one or more of N, O and S; the A is further replaced by R c Replaced by; said R c Substitution is one or more substitutions, when the substituent R c When there are multiple substituents, the substituents are the same or different; Ring C is a 6-10 membered aryl group or a 5-8 membered heteroaryl group; R a and R c Each is independently H, halogen, hydroxy, amino, cyano, carbonyl, oxo, C 1-6 Alkyl, C 3-8 Cycloalkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Halogenated alkyl, C 3-8 Halogenated cycloalkyl, C 1-6 Alkyl hydroxyl, C 1-6 Alkylcarbonyl, C 1-6 Alkoxy, C 1-6 Haloalkoxy or 3-8 membered heterocycloalkyl; R 1 、R 2 、R 3 are independently H, halogen, cyano, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Halogenated alkyl, C 3-8 Cycloalkyl, halogenated C 3-8 Cycloalkyl; R b Halogen 、 Hydroxyl, amino, cyano, carbonyl, oxo, C 1-6 Alkyl, C 1-6 Alkylamino, C 3-8 Cycloalkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Halogenated alkyl, C 3-8 Halogenated cycloalkyl, C 1-6 Alkyl hydroxyl, C 1-6 Alkylcarbonyl, C 1-6 Alkoxy, C 1-6 Halogenated alkoxy or 3-10 membered heterocycloalkyl, -(CHR j ) 0-3 -S(=O)(=NR j)-R k 、-(CHR j ) 1-3 -S(O)2-R k 、-(CHR j ) 1-3 -OC(O)-N(R k )2、-(CHR j ) 0-3 -(N=)S(=O)(R k )2、-(CHR j ) 0-3 -P(=O)(R k )2; said C 1-6 Alkyl, C 1-6 Alkylamino, C 3-8 Cycloalkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Halogenated alkyl, C 3-8 Halogenated cycloalkyl, C 1-6 Alkyl hydroxyl, C 1-6 Alkylcarbonyl, C 1-6 Alkoxy, C 1-6 Haloalkoxy, 3-10 membered heterocycloalkyl, -(CHR j ) 0-3 -S(=O)(=NR j )-R k 、-(CHR j ) 1-3 -S(O)2-R k 、-(CHR j ) 1-3 -OC(O)-N(R k )2、-(CHR j ) 0-3 -(N=)S(=O)(R k )2 and -(CHR j ) 0-3 -P(=O)(R k )2 is optionally replaced by one or more R d Substituted; said R d A substituent selected from the group consisting of halogen, hydroxy, amino, nitro, cyano, carbonyl, oxo, carboxyl, C 1-6 Alkyl, C 2-6 Alkynyl, C 1-6 Halogenated alkyl, C 1-6 Alkyl hydroxyl, C 1-6 Alkylcarbonyl, C 1-6 Alkoxy, C 1-6 Haloalkoxy; when the substituent R d When there are multiple R dare the same or different; n is 1, 2, 3 or 4, and R b At least one of -(CHR j ) 0-3 -S(=O)(=NR j )-R k 、-(CHR j ) 1-3 -S(O)2-R k 、-(CHR j ) 1-3 -OC(O)-N(R k )2、-(CHR j ) 0-3 -(N=)S(=O)(R k )2 or -(CHR j ) 0-3 -P(=O)(R k )2; R j and R k are independently H, cyano, amino, C 1-6 Alkyl, C 1-6 Alkylamino, -(CH2) 0-3 C 3-8 Cycloalkyl, -COC 1-6 Alkyl, C 1-6 Haloalkyl, C substituted by hydroxy 1-6 Alkyl, a single 5- to 10-membered heterocycloalkyl, or when the R k When there are two, they form a 3-10 membered heterocyclic alkyl group with the connected N, S or P atom; the amino, C 1-6 Alkylamino, -(CH2) 0-3 C 3-8 Cycloalkyl, -COC 1-6 Alkyl, C 1-6 Haloalkyl, C substituted by hydroxy 1-6 Alkyl, a single 5- to 10-membered heterocycloalkyl, and R k The 3-10 membered heterocycloalkyl formed with the attached N, S or P atom is optionally substituted by one or more R m Substituted; said R m A substituent selected from the group consisting of halogen, hydroxy, amino, nitro, cyano, carbonyl, oxo, carboxyl, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Alkyl hydroxyl, C 1-6 Alkylcarbonyl, C 1-6 Alkoxy, C 1-6 Halogenated alkoxy, C 3-8 Cycloalkyl; when the substituent R m When there are multiple R mSame or different; X 1 、X 2 、X 3 、X 4 、X 5 、X 6 , m and n are as defined in the first aspect of the present invention.

[0050] In a preferred embodiment of the present invention, the R b It is F, Cl, CN, -CH3, cyclopropyl, -CHF2, -CH2CF3, -NH-CH3, -CH2C(CH3)2(OH), -S(=O)(=NH)-CH3, -S(=O)(=NH)-CH2CH3, -S(=O)(=NH)- (CH2CH2OH), -CH2-S(=O)(=NH)-CH3, -CH2-S(=O)(=NCH3)-CH3, -CHCH3-S(=O)(=NCH3)-CH3, -C(CH3)2-S(=O)(=NCH3)-CH3, -S(=O)(=NCH3)-CH3, -S(=O)(=NCOCH3)-CH3, -S(=O)(=NCN)-CH3, -CH2-S(O)2-CH3, -C(C H3)2-S(O)2-CH3, -CH(CH2CH3)-S(O)2-CH3, -CH2-S(O)2-CH(CH3)2, -(CH2)2-S(O)2-CH3, -CH(CH3)-S(O)2-CH3, -CH(CH3)-S(O)2-CH2CH3, -CH2-S(O)2-CH3, -CH2-S(O)2-CH2CH3, -CH2CH(CH3)-S(O)2-CH3, -CH(CH3)CH2-S(O)2-CH3, -CH2-OC(O)-N(CH3)2, -(CH2)2-OC(O)-N(CH3)2, -CH2CH(CH3)-OC(O)-N(CH3)2, -CH(CH3)CH2-OC(O)-N(CH3)2, -(CH2)2-OC(O)-N(CH3)(CH2CH2CF3), -(CH2)2-OC(O)-N(CH3)(CH2CH2OH), -(N=)S(=O)(CH3)2、-(N=)S(=O)(CH2CH3)2、-CH2-(N=)S(=O)(CH3)2、 -P(=O)(CH3)2、-P(=O)(CH2CH3)2、

[0051] In a preferred embodiment of the present invention, the R b It is F, Cl, -CH3, -CH2CF3, -NH-CH3, -CH2C(CH3)2(OH), -S(=O)(=NH)-CH3, -S(=O)(=NH)-CH2CH3, -S(=O)(=NH)-(CH2CH 2OH), -CH2-S(=O)(=NH)-CH3, -CH2-S(=O)(=NCH3)-CH3, -CHCH3-S(=O)(=NCH3)-CH3, -C(CH3)2-S(=O)(=NCH3)-CH3, S(=O)(=NCH3)-CH3, -S(=O)(=NCOCH3)-CH3, -S(=O)(=NCN)-CH3, -CH2-S(O)2-CH3, -C(CH 3)2-S(O)2-CH3, -CH(CH2CH3)-S(O)2-CH3, -CH2-S(O)2-CH(CH3)2, -(CH2)2-S(O)2-CH3, -CH(CH3)-S(O)2-CH3, -CH2-S(O)2-CH3, -CH2-S(O)2-CH2CH3, -CH2CH(CH3)-S(O)2-CH3, -CH(CH3)CH2-S(O)2-CH3, -CH2-OC(O)-N(CH3)2, -(CH2)2-OC(O)-N(CH3)2, -CH2CH(CH3)-OC(O)-N(CH3)2, -CH(CH3)CH2-OC(O)-N(CH3)2, -(CH2)2-OC(O)-N(CH3)(CH2CH2CF3), -(CH2)2-OC(O)-N(CH3)(CH2CH2OH), -(N=)S(=O)(CH3)2、-(N=)S(=O)(CH2CH3)2、-CH2-(N=)S(=O)(CH3)2、 -P(=O)(CH3)2、-P(=O)(CH2CH3)2、

[0052] In a preferred embodiment of the present invention, the R j and R k Each is independently H, cyano, amino, -CH3, -CH2CH3, -CH2CH2CH3, -CH2CF3, -CH2CH2CF3, -NH-CH3, -COCH3, -CH(CH3)2, -CH2CH(CH3), -CH(CH3)CH2, -CH(CH3)-, -CH2CH2OH, Or the R k The N, S or P atoms to which it is attached form

[0053] In a preferred embodiment of the present invention, the R j and R k Each is independently H, cyano, amino, -CH3, -CH2CH3, -CH2CH2CH3, -CH2CF3, -CH2CH2CF3, -NH-CH3, -COCH3, -CH(CH3)2, -CH2CH(CH3), -CH(CH3)CH2, -CH(CH3)-, -CH2CH2OH, or the R k The N, S or P atoms to which it is attached form

[0054] In a preferred embodiment of the present invention, the compound shown in Formula I is selected from the following structures:

[0055] Among them, A and X 5 、X 6 The ring atoms together form a 6-8 membered aryl group, a 3-8 membered heterocycloalkyl group, a 5-membered heteroaromatic ring or a 6-membered heteroaromatic ring; the heteroatoms are independently selected from one or more of N, O and S; the A is further replaced by R c Replaced by; said R c Substitution is one or more substitutions, when the substituent R c When there are multiple substituents, the substituents are the same or different; Ring C is a 6-10 membered aryl group or a 5-8 membered heteroaryl group; R a and R c Each is independently H, halogen, hydroxy, amino, cyano, carbonyl, oxo, C 1-6 Alkyl, C 3-8 Cycloalkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Halogenated alkyl, C 3-8 Halogenated cycloalkyl, C 1-6 Alkyl hydroxyl, C 1-6 Alkylcarbonyl, C 1-6 Alkoxy, C 1-6 Haloalkoxy or 3-8 membered heterocycloalkyl; R 1 、R 2 、R 3are independently H, halogen, cyano, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Halogenated alkyl, C 3-8 Cycloalkyl, halogenated C 3-8 Cycloalkyl; R b Halogen 、 Hydroxyl, amino, cyano, carbonyl, oxo, C 1-6 Alkyl, C 3-8 Cycloalkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Halogenated alkyl, C 3-8 Halogenated cycloalkyl, C 1-6 Alkyl hydroxyl, C 1-6 Alkylcarbonyl, C 1-6 Alkoxy, C 1-6 Halogenated alkoxy or 3-10 membered heterocycloalkyl, -(CHR j ) 0-3 -S(=O)(=NR j )-R k 、-(CHR j ) 1-3 -S(O)2-R k 、-(CHR j ) 1-3 -OC(O)-N(R k )2、-(CHR j ) 0-3 -(N=)S(=O)(R k )2、-(CHR j ) 0-3 -P(=O)(R k )2; said C 1-6 Alkyl, C 3-8 Cycloalkyl, C 2-6 Alkynyl, C 1-6 Halogenated alkyl, C 3- 8-halogenated cycloalkyl, C 1-6 Alkyl hydroxyl, C 1-6 Alkylcarbonyl, C 1-6 Alkoxy and C 1-6 The haloalkoxy group is optionally substituted with one or more R d Substituted; said R d A substituent selected from the group consisting of halogen, hydroxy, amino, nitro, cyano, carbonyl, oxo, carboxyl, C 1-6 Alkyl, C 2-6 Alkynyl, C 1- 6 haloalkyl, C 1-6 Alkyl hydroxyl, C 1-6 Alkylcarbonyl, C1-6 Alkoxy, C 1-6 Haloalkoxy; when the substituent R d When there are multiple R d are the same or different; n is 1, 2, 3 or 4, and R b At least one of -(CHR j ) 0-3 -S(=O)(=NR j )-R k 、-(CHR j ) 1-3 -S(O)2-R k 、-(CHR j ) 1-3 -OC(O)-N(R k )2、-(CHR j ) 0-3 -(N=)S(=O)(R k )2 or -(CHR j ) 0-3 -P(=O)(R k )2; R j and R k are independently H, cyano, amino, C 1-6 Alkyl, C 3-8 Cycloalkyl, C 1-6 Haloalkyl, single 5- to 8-membered heterocycloalkyl, or when the R k When there are two, they form a 5-8 membered heterocyclic alkyl group with the connected N, S or P atom; the amino, C 1-6 Alkyl, C 3-8 Cycloalkyl, C 1-6 haloalkyl, a single 5- to 8-membered heterocycloalkyl, and R k The 5- to 8-membered heterocycloalkyl formed with the attached N, S or P atom is optionally substituted with one or more R m Substituted; said R m A substituent selected from the group consisting of halogen, hydroxy, amino, nitro, cyano, carbonyl, oxo, carboxyl, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Alkyl hydroxyl, C 1-6 Alkylcarbonyl, C 1-6 Alkoxy, C 1-6 Halogenated alkoxy, C 3-8 Cycloalkyl; when the substituent R m When there are multiple R m Same or different; X 1 、X 2 、X 3 、X 4 、X 5、X 6 , m and n are as defined in the second aspect of the present invention.

[0056] In a preferred embodiment of the present invention, Selected from

[0057] In a preferred embodiment of the present invention, for

[0058] In a preferred embodiment of the present invention, Ring B is For example

[0059] In a preferred embodiment of the present invention, the R b It is F, Cl, -S(=O)(=NH)-CH3, -S(=O)(=NH)-CH2CH3, -S(=O)(=NH)-(CH2CH2OH), -CH2-S(=O)(=NH)-CH3, -S(=O)(=NCH3)-CH3, -S(=O)(=NCOCH3)-CH3, -S(=O)(=NCN)-CH3, -CH2-S(O)2-CH3, -(CH2)2-S(O)2-CH3, -CH2CH(CH3)-S(O)2-CH3, -CH(CH3)CH2-S(O)2-CH3, -(CH2)2-OC(O)-N(CH3)2, -CH2CH(CH3)-OC(O)-N(CH3)2, -CH(CH3)CH2-OC(O)-N(CH3)2, -(CH2)2-OC(O)-N(CH3)(CH2CH2CF3), -(CH2)2-OC(O)-N(CH3)(CH2CH2OH), -(N=)S(=O)(CH3)2、-(N=)S(=O)(CH2CH3)2、-CH2-(N=)S(=O)(CH3)2、 -P(=O)(CH3)2、-P(=O)(CH2CH3)2、

[0060] In a preferred embodiment of the present invention, R d It is a methyl group.

[0061] In a preferred embodiment of the present invention, the R a 、R c Each independently represents H, F, Cl, methyl, amino, oxo, -CF3, -CHF2, -CN。

[0062] In a preferred embodiment of the present invention, the R a 、R c Each is independently H, F, Cl, methyl, amino, or oxo.

[0063] In a preferred embodiment of the present invention, the R 1 、R 2 、R 3 Each independently represents H, methyl, -CH2CF3, -CF3,

[0064] In a preferred embodiment of the present invention, ring C is a 6- to 8-membered aryl group or a 5- to 8-membered heteroaryl group.

[0065] In a preferred embodiment of the present invention, ring C is a 5- to 6-membered nitrogen-containing heteroaryl group, and the number of the nitrogen atoms is 1, 2 or 3.

[0066] In a preferred embodiment of the present invention, ring C is phenyl, pyrazolyl or pyridinyl.

[0067] In a preferred embodiment of the present invention, ring C is

[0068] In a preferred embodiment of the present invention, for

[0069] In a preferred embodiment of the present invention, the compound is selected from the following structures:

[0070] Among them, R 1 、R 2 、R 3 、R b 、R c , n independently have the definitions given above.

[0071] In a preferred embodiment of the present invention, the compound is selected from the following structures:

[0072] Among them, R 1 、R 2 、R 3 、R b 、R c , n independently have the definitions given above.

[0073] In a preferred embodiment of the present invention, the compound represented by formula IA is selected from any one of the following compounds:

[0074] In a preferred embodiment of the present invention, the pharmaceutically acceptable salt of the compound of Formula IA is trifluoroacetate.

[0075] In a preferred embodiment of the present invention, the compound represented by formula IA is selected from any one of the following compounds:

[0076] In a preferred embodiment of the present invention, the compound represented by formula IA is selected from any one of the following compounds:

[0077] The third aspect of the present invention provides a pharmaceutical composition, which comprises: the compound as described in the first to second aspects of the present invention, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug; and a pharmaceutically acceptable carrier.

[0078] The fourth aspect of the present invention provides the use of the compound as described in the first to second aspects of the present invention, its tautomers, stereoisomers, hydrates, solvates, pharmaceutically acceptable salts or prodrugs, or the use of the pharmaceutical composition as described in the third aspect of the present invention, the uses including: antagonizing MRGPRX2; and / or preventing and / or treating MRGPRX2-related diseases; and / or preparing drugs, pharmaceutical compositions or preparations for antagonizing MRGPRX2, and / or preventing and / or treating MRGPRX2-related diseases.

[0079] Preferably, the MRGPRX2-related diseases include mast cell-related diseases.

[0080] Preferably, the MRGPRX2-related diseases include: skin diseases, autoimmune diseases, and nervous system diseases.

[0081] Preferably, the skin disease is selected from atopic dermatitis, contact dermatitis, urticaria, and pruritus.

[0082] Preferably, the urticaria is selected from chronic spontaneous urticaria and induced urticaria.

[0083] Preferably, the autoimmune disease is selected from the group consisting of allergy, mastocytosis, rheumatoid arthritis, asthma, ulcerative colitis and interstitial cystitis.

[0084] Preferably, the neurological disease is selected from pain.

[0085] In the fifth aspect of the present invention, a method for antagonizing MRGPRX2, or preventing and / or treating MRGPRX2-related diseases is provided, comprising the steps of administering to a subject in need thereof the compound described in the first to second aspects of the present invention, its tautomers, stereoisomers, hydrates, solvates, pharmaceutically acceptable salts or prodrugs.

[0086] Additional aspects and advantages of the present invention will be set forth in part in the description which follows and, in part, will be obvious from the description which follows, or may be learned by practice of the present invention.

[0087] Definitions and Explanations of Terms

[0088] Unless otherwise specified, the terms and definitions used in this application, including the description and claims, are as follows. It will be understood by those skilled in the art that, according to the conventions used in the art, in the structural formula of this application, Used to depict chemical bonds, which are the points where a moiety or substituent is attached to a core or backbone structure.

[0089] Unless otherwise specified, the term "pharmaceutically acceptable" refers to those compounds, materials, compositions and / or dosage forms which, within the scope of sound medical judgment, are suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problems or complications, commensurate with a reasonable benefit / risk ratio.

[0090] Unless otherwise specified, the term "pharmaceutically acceptable salts" refers to salts of pharmaceutically acceptable non-toxic acids or bases including salts of inorganic acids and bases, and organic acids and bases.

[0091] Unless otherwise specified, the term "pharmaceutical composition" means a mixture of one or more compounds described herein, or their physiologically / pharmaceutically acceptable salts or prodrugs, with other chemical components, such as physiologically / pharmaceutically acceptable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate administration of the compound to an organism.

[0092] Unless otherwise specified, the term "prodrug" refers to a compound of the present invention that can be converted to a biologically active compound under physiological conditions or by solvolysis. Prodrugs of the present invention are prepared by modifying functional groups within the compound. These modifications can be removed by conventional procedures or in vivo to yield the parent compound. Prodrugs include compounds in which a hydroxyl or amino group within a compound of the present invention is attached to any group. When a prodrug of a compound of the present invention is administered to a mammalian subject, the prodrug is cleaved to form a free hydroxyl group or free amino group, respectively.

[0093] Unless otherwise specified, the term "stereoisomer" refers to isomers resulting from different arrangements of atoms in a molecule in space, and includes cis-trans isomers, enantiomers, diastereomers, and conformational isomers.

[0094] Depending on the choice of raw materials and methods, the compounds of the present invention may exist in the form of one of the possible isomers or a mixture thereof, for example as pure optical isomers, or as a mixture of isomers, such as a racemic and diastereomeric mixture, depending on the number of asymmetric carbon atoms. When describing an optically active compound, the prefixes D and L or R and S are used to indicate the absolute configuration of the molecule with respect to the chiral center (or multiple chiral centers) in the molecule. The prefixes D and L or (+) and (–) are the symbols used to specify the rotation of plane polarized light caused by the compound, where (–) or L indicates that the compound is left-handed. Compounds prefixed with (+) or D are right-handed. With respect to a given chemical structure, these stereoisomers are identical except that they are mirror images of each other. Specific stereoisomers may also be referred to as enantiomers, and mixtures of the isomers are often referred to as mixtures of enantiomers. A 50:50 mixture of enantiomers is called a racemic mixture or racemate, which can occur when there is no stereoselectivity or stereospecificity in a chemical reaction or process. Many geometric isomers of alkenes, C=N double bonds, etc. can also exist in the compounds described herein, and all such stable isomers are contemplated by the present invention. When the compounds described herein contain olefinic double bonds, unless otherwise specified, such double bonds include both E and Z geometric isomers. If the compound contains a disubstituted cycloalkyl group, the cycloalkyl substituents may be in either the cis- or trans- configuration.

[0095] When bonds to chiral carbon atoms in formulae of the present invention are depicted as straight lines, it is understood that both the (R) and (S) configurations of the chiral carbon atoms and the enantiomerically pure compounds and mixtures thereof are encompassed within the scope of the formulae. The diagrammatic representations of racemates and enantiomerically pure compounds herein are adapted from Maehr, J. Chem. Ed. 1985, 62: 114-120. Unless otherwise indicated, wedge-shaped bonds and dashed bonds are used to represent the absolute configuration of a stereocenter.

[0096] Optically active (R)- or (S)-isomers can be prepared using chiral synthons or chiral preparations, or resolved using conventional techniques. Compounds of the invention containing asymmetrically substituted carbon atoms can be separated in optically active form or racemic form. Resolution of a racemic mixture of a compound can be carried out by any of a number of methods known in the art. An exemplary method includes fractional recrystallization using a chiral resolving acid that is an optically active, salified organic acid. Suitable resolving agents for fractional recrystallization methods are, for example, optically active acids, such as tartaric acid, diacetyltartaric acid, dibenzoyltartaric acid, mandelic acid, malic acid, lactic acid, or various optically active camphorsulfonic acids such as the D and L forms of β-camphorsulfonic acid. Other resolving agents suitable for fractional crystallization methods include α-methyl-benzylamine (e.g., S and R forms or diastereoisomerically pure forms), 2-phenylglycinol, norephedrine, ephedrine, N-methylephedrine, cyclohexylethylamine, 1,2-diaminocyclohexane, etc. The resolution of the racemic mixture can also be carried out by eluting on a chromatographic column filled with an optically active resolving agent (e.g., dinitrobenzoylphenylglycine). High performance liquid chromatography (HPLC) can also be used to carry out supercritical fluid chromatography (SFC). The selection of specific methods and elution conditions, chromatographic column selection can be selected by those skilled in the art according to the structure of the compound and test results. Further, optically pure starting materials or reagents of known configuration can also be used to obtain any enantiomer or diastereomer of the compound described in the present invention through stereoorganic synthesis.

[0097] Unless otherwise specified, the term "tautomer" refers to functional group isomers resulting from the rapid shift of an atom in a molecule between two positions. The compounds of the present invention may exhibit tautomerism. Tautomeric compounds may exist as two or more interconvertible species. Prototropic tautomers arise from the migration of a covalently bonded hydrogen atom between two atoms. Tautomers generally exist in equilibrium, and attempts to isolate a single tautomer usually produce a mixture whose physical and chemical properties are consistent with a mixture of compounds. The position of equilibrium depends on the chemical properties within the molecule. For example, in many aliphatic aldehydes and ketones such as acetaldehyde, the keto form predominates, while in phenols, the enol form predominates. The present invention encompasses all tautomeric forms of the compounds.

[0098] Unless otherwise specified, use a solid wedge key. and dotted wedge key To indicate the absolute configuration of a stereocenter, use a straight solid bond and straight dashed key Indicates the relative configuration of a stereocenter.

[0099] Unless otherwise indicated, the term "solvate" means that the compound of the present invention or its salt includes a stoichiometric or non-stoichiometric amount of a solvent bound by non-covalent intermolecular forces. When the solvent is water, it is a hydrate.

[0100] With respect to a drug or pharmacologically active agent, the term "effective amount" or "therapeutically effective amount" refers to a non-toxic amount of the drug or agent sufficient to achieve the intended effect. For the oral dosage forms of the present invention, an "effective amount" of an active substance in the composition means the amount required to achieve the intended effect when used in combination with another active substance in the composition. The determination of an effective amount varies from person to person, depending on the age and general condition of the recipient, as well as the specific active substance. The appropriate effective amount in each individual case can be determined by those skilled in the art through routine experimentation.

[0101] Unless otherwise specified, the terms "active ingredient," "therapeutic agent," "active substance," or "active agent" refer to a chemical entity that is effective in treating a target disorder, disease, or condition.

[0102] Unless otherwise specified, the term "substituted" means that any one or more hydrogen atoms on a particular atom are replaced by a substituent, including deuterium and hydrogen variants, as long as the valence state of the particular atom is normal and the substituted compound is stable. When the substituent is a keto group (i.e., =0), it means that two hydrogen atoms are replaced. Keto substitution does not occur on aromatic groups.

[0103] In this application, "optional" or "optionally" means that the subsequently described event or circumstance may or may not occur, and the description includes both instances where the event or circumstance occurs and instances where it does not occur. For example, "optionally substituted aryl" means that the aryl group is substituted or unsubstituted, and the description includes both substituted aryl groups and unsubstituted aryl groups.

[0104] Unless otherwise specified, the term “C 1-6 "Alkyl" is used to represent a straight or branched chain saturated hydrocarbon group consisting of 1 to 6 carbon atoms. 1- 6 alkyl groups including C 1-5 、C 1-4 、C 1-3 、C 1-2 、C 2-6 、C 2-4 , C6 and C5 alkyl, etc.; which can be monovalent (such as methyl), divalent (such as methylene) or polyvalent (such as methine). 1-6 Examples of alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), butyl (including n-butyl, isobutyl, s-butyl and t-butyl), pentyl (including n-pentyl, isopentyl and neopentyl), hexyl, and the like.

[0105] Unless otherwise specified, the term “C 1-3 "Alkyl" is used to represent a straight or branched chain saturated hydrocarbon group consisting of 1 to 3 carbon atoms. 1- 3 alkyl groups including C 1-2 and C 2-3 Alkyl, etc.; it can be monovalent (such as methyl), divalent (such as methylene) or polyvalent (such as methine). 1-3 Examples of alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), and the like.

[0106] The term "halo" by itself or as part of another substituent is used interchangeably with the term "halogen-substituted."

[0107] Unless otherwise specified, "haloalkyl" or "halogen-substituted alkyl" refers to a saturated aliphatic hydrocarbon group including branched and straight chains having the specified number of carbon atoms, substituted by one or more halogens, which may be "C 1-6 "Haloalkyl".

[0108] Unless otherwise specified, “C 2-6 "Alkenyl" is used to refer to a linear or branched hydrocarbon group consisting of 2 to 6 carbon atoms containing at least one carbon-carbon double bond, which may be located at any position of the group. 2-6 Alkenyl groups include C 2-4 、C 2-3 , C4, C3 and C2 alkenyl, etc.; which may be monovalent, divalent or polyvalent. 2-6 Examples of alkenyl groups include, but are not limited to, ethenyl, propenyl, butenyl, pentenyl, hexenyl, butadienyl, piperyl, hexadienyl, and the like.

[0109] Unless otherwise specified, “C 2-6 "Alkynyl" is used to represent a linear or branched hydrocarbon group consisting of 2 to 6 carbon atoms containing at least one carbon-carbon triple bond, which may be located at any position of the group. 2-6 Alkynyl groups include C 2-4 、C 2-3 , C4, C3 and C2 alkynyl, etc. It can be monovalent, divalent or polyvalent. 2-6 Examples of alkynyl groups include, but are not limited to, ethynyl, propynyl, butynyl, pentynyl, and the like.

[0110] Unless otherwise specified, the term “C 1-6 "Alkoxy" refers to an alkyl group containing 1 to 6 carbon atoms which is attached to the rest of the molecule via an oxygen atom. 1-6 Alkoxy groups include C 1-4 、C 1-3 、C 1-2、C 2-6 、C 2-4 , C6, C5, C4 and C3 alkoxy, etc. 1-6 Examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy (including n-propoxy and isopropoxy), butoxy (including n-butoxy, isobutoxy, s-butoxy and t-butoxy), pentoxy (including n-pentoxy, isopentoxy and neopentoxy), hexyloxy, and the like.

[0111] Unless otherwise specified, the term “C 1-3 "Alkoxy" refers to those alkyl groups containing 1 to 3 carbon atoms which are attached to the rest of the molecule via an oxygen atom. The C 1-3 Alkoxy groups include C 1-2 、C 2-3 , C3 and C2 alkoxy, etc. 1-3 Examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy (including n-propoxy and isopropoxy), and the like.

[0112] Unless otherwise specified, the term “C 1-6 "Alkylamino" means an alkyl group containing 1 to 6 carbon atoms which is attached to the rest of the molecule via an amino group. 1-6 Alkylamino groups include C 1-4 、C 1-3 、C 1-2 、C 2-6 、C 2-4 , C6, C5, C4, C3 and C2 alkylamino, etc. 1-6 Examples of alkylamino groups include, but are not limited to, -NHCH3, -N(CH3)2, -NHCH2CH3, -N(CH3)CH2CH3, -N(CH2CH3)(CH2CH3), -NHCH2CH2CH3, -NHCH2(CH3)2, -NHCH2CH2CH2CH3, and the like.

[0113] Unless otherwise specified, the term “C 3-12 "Cycloalkyl" refers to a saturated cyclic hydrocarbon group consisting of 3 to 12 carbon atoms, including monocyclic and bicyclic systems, wherein the bicyclic system includes spirocyclic, fused and bridged rings. 3-12 Cycloalkyl groups include C 3-8 、C 3-6 、C 3-5 、C 4-8 、C 4-6 、C 4-5 、C 5- 8 or C 5-6 Cycloalkyl, etc.; it may be monovalent, divalent or polyvalent. 3-12Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, norbornyl, [2.2.2]bicyclooctane, and the like.

[0114] Unless otherwise specified, the term “C 3-6 "Cycloalkyl" means a saturated cyclic hydrocarbon group consisting of 3 to 6 carbon atoms, which is a monocyclic or bicyclic ring system. 3-6 Cycloalkyl groups include C 3-5 、C 4-5 and C 5-6 Cycloalkyl, etc.; it may be monovalent, divalent or polyvalent. 3-6 Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like.

[0115] Unless otherwise specified, C n-n+m or C n -C n+m Any specific case including n to n+m carbons, such as C 1-12 Including C1, C2, C3, C4, C5, C6, C7, C8, C9, C 10 、C 11 , and C 12 , also includes any range from n to n+m, such as C 1-12 Including C 1-3 、C 1- 6. C 1-9 、C 3-6 、C 3-9 、C 3-12 、C 6-9 、C 6-12 , and C 9-12 Similarly, n-membered to n+m-membered means that the number of atoms in the ring is n to n+m, for example, a 3-12-membered ring includes a 3-membered ring, a 4-membered ring, a 5-membered ring, a 6-membered ring, a 7-membered ring, an 8-membered ring, a 9-membered ring, a 10-membered ring, an 11-membered ring, and a 12-membered ring, and also includes any range from n to n+m, for example, a 3-12-membered ring includes a 3-6-membered ring, a 3-9-membered ring, a 5-6-membered ring, a 5-7-membered ring, a 6-7-membered ring, a 6-8-membered ring, and a 6-10-membered ring, etc.

[0116] Unless otherwise specified, the term "aryl" refers to a monocyclic or polycyclic carbocyclic ring having 6 to 20 carbon atoms, at least one of which is aromatic. When one of the rings is non-aromatic, the group may be attached via either the aromatic or non-aromatic ring. Examples of aryl groups include, but are not limited to, phenyl, naphthyl, tetrahydronaphthyl, indanyl, biphenyl, phenanthrenyl, anthracenyl, and acenaphthenyl, and may be a "6- to 10-membered aryl" or a "6- to 8-membered aryl." The term "6- to 10-membered aryl" refers to a monocyclic or polycyclic carbocyclic ring having 6 to 10 carbon atoms, at least one of which is aromatic.

[0117] Unless otherwise specified, the term "heterocycloalkyl" refers to a cycloalkyl group in which one or more (in some embodiments, 1 to 3) carbon atoms are replaced by heteroatoms such as, but not limited to, N, O, S, and P. The term "mn-membered heterocycloalkyl" or "Cm-Cn heterocycloalkyl" is understood to mean a saturated, unsaturated or partially saturated ring having m to n atoms, wherein the heteroatoms are selected from N, O, S, P, preferably from N, O or S, and may be "3-11-membered heterocycloalkyl, 3-10-membered heterocycloalkyl, 5-11-membered heterocycloalkyl, 5-10-membered heterocycloalkyl, 3-8-membered heterocycloalkyl, 5-8-membered heterocycloalkyl. The term "3-11-membered heterocycloalkyl" alone or in combination with other terms means a saturated cyclic group consisting of 3 to 11 ring atoms. The term "6-8-membered heterocycloalkyl" alone or in combination with other terms means a saturated cyclic group consisting of 6 to 8 ring atoms, 1, 2, 3 or 4 of which are heteroatoms independently selected from O, S and N, and the rest are carbon atoms, wherein the nitrogen atom is optionally quaternized, and the nitrogen and sulfur heteroatoms may be optionally oxidized (i.e., NO and S(O)p, where p is 1 or 2 ). It includes monocyclic and bicyclic ring systems, wherein the bicyclic ring system includes spirocyclic, cyclic and bridged rings. In addition, with respect to the "6-8 membered heterocycloalkyl", a heteroatom may occupy the position where the heterocycloalkyl is connected to the rest of the molecule. For example, a 6-8 membered heterocycloalkyl includes but is not limited to 6-membered, 7-membered, and 8-membered. Examples of 6-8 membered heterocycloalkyls include but are not limited to azetidinyl, oxetanyl, thietanyl, pyrrolidinyl, pyrazolidinyl, imidazolidinyl, tetrahydrothiophenyl (including tetrahydrothiophene-2-yl and tetrahydrothiophene-3-yl, etc.), tetrahydrofuranyl (including tetrahydrofuran-2-yl, etc.), tetrahydropyranyl, piperidinyl (including 1-piperidinyl, 2-piperidinyl and 3-piperidinyl, etc.), piperazinyl (including 1-piperazinyl and 2-piperazinyl, etc.), morpholinyl (including 3-morpholinyl and 4-morpholinyl, etc.), dioxanyl, dithianyl, isoxazolidinyl, isothiazolidinyl, 1,2-oxazinyl, 1,2-thiazinyl, hexahydropyridazinyl.

[0118] Unless otherwise specified, the term "heteroaromatic ring" refers to a monocyclic or polycyclic carbon ring having 5 to 10 atoms, wherein at least one ring atom is a heteroatom independently selected from oxygen, sulfur and nitrogen, and the remaining ring atoms are C, wherein at least one ring is an aromatic ring. The group may be a carbon group or a heteroatom group (i.e., it may be C-connected or N-connected, as long as it is possible). When one of the rings is a non-aromatic ring, the group may be connected through the aromatic ring or through the non-aromatic ring. Examples of heteroaryl groups include, but are not limited to, imidazolyl, acridinyl, carbazolyl, cinnolinyl, quinoxalinyl, pyrazolyl, indolyl, benzotriazolyl, furyl, thienyl, benzothienyl, benzofuranyl, quinolyl, isoquinolyl, oxazolyl, isoxazolyl, indolyl, pyrazinyl, pyridazinyl, pyridyl, pyrimidinyl, pyrrolyl, N-methylpyrrolyl and tetrahydroquinoline. The term "heteroaromatic ring" can be used interchangeably with the term "heteroaromatic ring", "heteroaryl" or "heteroaromatic ring group", and can refer to "5- to 10-membered heteroaryl group, 5- to 8-membered heteroaryl group".

[0119] Unless otherwise specified, the term "oxo" refers to the replacement of two hydrogen atoms on a methylene group by oxygen atoms, ie, the methylene group is replaced by a carbonyl group, representing =0.

[0120] Unless otherwise specified, the term "halo" or "halogen" refers to fluoro, chloro, bromo and iodo.

[0121] Additionally, it should be noted that, unless otherwise expressly stated, the term "independently" used in the present invention should be broadly construed to mean that the individual entities described are independent of one another and may independently represent the same or different specific groups. More specifically, the term "independently" can mean that specific options expressed by identical symbols in different groups do not affect each other, or that specific options expressed by identical symbols in the same group do not affect each other.

[0122] In this application, "pharmaceutically acceptable carrier" includes but is not limited to any adjuvant, carrier, excipient, glidant, sweetener, diluent, preservative, dye / colorant, flavoring, surfactant, wetting agent, dispersant, suspending agent, stabilizer, isotonic agent, solvent or emulsifier approved by the relevant governmental regulatory authorities as acceptable for human or livestock use.

[0123] The term "treat" refers to therapeutic treatment. When referring to a specific condition, treatment means: (1) alleviating the disease or one or more biological manifestations of the condition, (2) interfering with (a) one or more points in the biological cascade that leads to or causes the condition or (b) one or more biological manifestations of the condition, (3) ameliorating one or more symptoms, effects, or side effects associated with the condition or one or more symptoms, effects, or side effects associated with the condition or its treatment, or (4) slowing the progression of the condition or one or more biological manifestations of the condition.

[0124] The term "prevent" refers to the reduction of the risk of acquiring or developing a disease or disorder.

[0125] The term "patient" refers to any animal, preferably a mammal, that is about to be or has been administered a compound or composition according to embodiments of the present invention. The term "mammal" includes any mammal. Examples of mammals include, but are not limited to, cattle, horses, sheep, pigs, cats, dogs, mice, rats, rabbits, guinea pigs, monkeys, humans, etc., preferably humans.

[0126] The term "therapeutically effective amount" refers to an amount of a compound that, when administered to a patient, is sufficient to effectively treat a disease or condition described herein. The "therapeutically effective amount" will vary depending on the compound, the condition and its severity, and the age of the patient to be treated, and can be adjusted as needed by those skilled in the art.

[0127] Without violating the common sense in the art, the above-mentioned preferred conditions can be arbitrarily combined to obtain preferred embodiments of the present invention. Beneficial effects

[0128] After extensive and in-depth research, the inventors unexpectedly discovered a compound or pharmaceutically acceptable salt thereof, as well as a preparation method and use thereof. The present invention provides a compound represented by Formula IA, and its tautomers, stereoisomers, hydrates, solvates, pharmaceutically acceptable salts, or prodrugs. The compound of Formula IA exhibits significant inhibitory effects on MRGPRX2 and can be used as a MAS-related G protein receptor X2 antagonist, exhibiting high safety and druggable properties. DETAILED DESCRIPTION

[0129] The present invention will be further described below in conjunction with specific examples. It should be understood that the following description is only the most preferred embodiment of the present invention and should not be considered as limiting the scope of protection of the present invention. Based on a full understanding of the present invention, the experimental methods in the following examples that do not specify specific conditions are generally carried out under conventional conditions or under conditions recommended by the manufacturer. Those skilled in the art may make non-essential changes to the technical solutions of the present invention, and such changes should be considered as included in the scope of protection of the present invention.

[0130] This application has the following definitions:

[0131] Symbol or unit:

[0132] IC 50 : Half-maximal inhibitory concentration, which refers to the concentration at which half of the maximum inhibitory effect is achieved

[0133] M: mol / L, for example, n-butyllithium (14.56 mL, 29.1 mmol, 2.5 M n-hexane solution) means a n-butyllithium n-hexane solution with a molar concentration of 2.5 mol / L

[0134] N: equivalent concentration, for example, 2N hydrochloric acid means 2 mol / L hydrochloric acid solution

[0135] Reagents:

[0136] PPA: polyphosphoric acid

[0137] DCE: 1,2-dichloroethane

[0138] Intermediate A1: (1S,3R)-N 1 Synthesis of 6-chloro-2-(trifluoromethyl)quinolin-4-yl)cyclohexane-1,3-diamine

[0139] The synthetic route of intermediate A1 is as follows:

[0140] Step 1: Synthesis of 6-chloro-2-(trifluoromethyl)quinolin-4-ol (A1-3)

[0141] At room temperature, p-chloroaniline (5 g, 39.37 mmol) was added to PPA (200 mmol, 75 mL, 5.0 eq), followed by ethyl trifluoroacetate (7.2 g, 39.37 mmol). The reaction mixture was heated to 140°C for 12 hours. After completion, the reaction mixture was poured into ice water and extracted with ethyl acetate (4 x 50 mL). The resulting organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to afford the crude compound 6-chloro-2-(trifluoromethyl)quinolin-4-ol (8.5 g, 87.6% yield), which was used directly in the next reaction.

[0142] LC-MS, M / Z(ESI):248.0[M+H] +

[0143] Step 2: Synthesis of 4,6-dichloro-2-(trifluoromethyl)quinoline (A1-4)

[0144] 6-Chloro-2-(trifluoromethyl)quinolin-4-ol (A1-3) (8.5 g, 34.5 mmol) was dissolved in DCE (100 mL) and phosphorus oxychloride (13.2 g, 86.25 mmol) was added at 0°C. The reaction temperature was then raised to 80°C for 12 hours. After completion of the reaction, the crude product was dried and purified via a silica gel column (PE:EA (V / V) = 100:2) to afford 4,6-dichloro-2-(trifluoromethyl)quinoline (8.3 g, 91.6% yield).

[0145] LC-MS, M / Z(ESI):265.9[M+H] +

[0146] Step 3: Synthesis of tert-butyl ((1R,3S)-3-((6-chloro-2-(trifluoromethyl)quinolin-4-yl)-amino)cyclohexane)carbamate (A1-6)

[0147] At room temperature, 4,6-dichloro-2-(trifluoromethyl)quinoline (A1-4) (1.32 g, 5 mmol) and tert-butyl (1R,3S)-3-aminocyclohexane (A1-5) (1 g, 4.67 mmol) were added to N-methylpyrrolidone (10 mL), followed by N,N-diisopropylethylamine (1.9 g, 15 mmol). The reaction mixture was heated to 130°C for 12 hours. After completion of the reaction, the reaction mixture was poured into ice water and extracted with ethyl acetate (4 x 50 mL). The resulting organic phase was dried over anhydrous sodium sulfate, filtered, and dried. The crude product was purified on a silica gel column (PE:EA (V / V) = 2:1) to afford tert-butyl ((1R,3S)-3-((6-chloro-2-(trifluoromethyl)quinolin-4-yl)-amino)cyclohexane)carbamate (A1-6) (1.9 g, 96.3% yield).

[0148] LC-MS, M / Z(ESI):444.1[M+H] +

[0149] Step 4: (1S, 3R)-N 1 Synthesis of -(6-chloro-2-(trifluoromethyl)quinolin-4-yl)cyclohexane-1,3-diamine (A1)

[0150] At room temperature, tert-butyl ((1R,3S)-3-((6-chloro-2-(trifluoromethyl)quinolin-4-yl)-amino)cyclohexane)carbamate (A1-6) (1 g, 2.3 mmol) was dissolved in 1,4-dioxane (20 mL), and concentrated hydrochloric acid (0.5 mL) was added. The reaction solution was stirred at room temperature for 1 hour. After the reaction was completed, saturated sodium bicarbonate aqueous solution was added to the reaction solution to make it alkaline, and the mixture was extracted with dichloromethane / methanol (4*50 mL). The obtained organic phase was dried over anhydrous sodium sulfate, filtered and dried, and the crude product was purified by silica gel column chromatography (DCM:MeOH (V / V) = 100:2) to obtain compound (1S,3R)-N 1 -(6-chloro-2-(trifluoromethyl)quinolin-4-yl)cyclohexane-1,3-diamine (720 mg, 91.3% yield).

[0151] LC / MS(ESI)(m / z):344[M+H] +

[0152] Example 1: Preparation of N-((1R,3S)-3-((6-chloro-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)-1-(2-(methylsulfonyl)ethyl)-1H-pyrazole-4-carboxamide trifluoroacetate (Compound 1)

[0153] The synthetic route of compound 1 is as follows:

[0154] Step 1: Synthesis of ethyl 1-(2-(methylsulfonyl)ethyl)-1H-pyrazole-4-carboxylate

[0155] To a solution of ethyl 1H-pyrazole-4-carboxylate (500 mg, 3.57 mmol) in N,N-dimethylformamide (5 mL) was added 2-bromoethyl methyl sulfone (801 mg, 4.28 mmol), potassium carbonate (986 mg, 7.14 mmol), and potassium iodide (118 mg, 0.71 mmol). The resulting mixture was then stirred at 25°C for 3 hours. The mixture was diluted with water (20 mL) and extracted with ethyl acetate (15 mL x 3). The organic layers were combined, washed with saturated brine (30 mL), dried over sodium sulfate, and concentrated to obtain the crude product. The product was separated and purified on a silica gel column (methanol:dichloromethane (V / V) = 100:1-100:5, gradient elution) to obtain ethyl 1-(2-(methylsulfonyl)ethyl)-1H-pyrazole-4-carboxylate (300 mg, 34.1% yield).

[0156] LC-MS, M / Z(ESI):247.2[M+H] +

[0157] Step 2: Synthesis of 1-(2-(methylsulfonyl)ethyl)-1H-pyrazole-4-carboxylic acid

[0158] To a solution of ethyl 1-(2-(methylsulfonyl)ethyl)-1H-pyrazole-4-carboxylate (300 mg, 1.22 mmol) in tetrahydrofuran (2 mL) and water (2 mL) was added lithium hydroxide (51.1 mg, 1.22 mmol), and the resulting mixture was stirred at 25° C. for 18 hours. The reaction solution was concentrated by rotary evaporation, and the crude product was separated and purified using a reverse phase column (water:acetonitrile (V / V) = 100:0-100:10) to obtain 1-(2-(methylsulfonyl)ethyl)-1H-pyrazole-4-carboxylic acid (230 mg, 86.5% yield).

[0159] LC-MS, M / Z(ESI):219.2[M+H] +

[0160] Step 3: N-((1R,3S)-3-((6-chloro-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)-1-(2-(methylsulfonyl)ethyl)-1H-pyrazole-4-carboxamide trifluoroacetate (Compound 1)

[0161] To (1S,3R)-N 1 To a solution of -(6-chloro-2-(trifluoromethyl)quinolin-4-yl)cyclohexane-1,3-diamine (Intermediate A1) (120 mg, 0.35 mmol) in N,N-dimethylformamide (2 mL) were added 1-(2-(methylsulfonyl)ethyl)-1H-pyrazole-4-carboxylic acid (114 mg, 0.52 mmol), N,N-diisopropylethylamine (0.17 mL, 1.05 mmol) and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (199 mg, 0.52 mmol), and the resulting mixture was stirred at 25 ° C under nitrogen protection for 2 hours. The reaction solution was concentrated under reduced pressure and then purified by preparative high performance liquid chromatography (chromatographic column: Kinetex EVO C18, 21.2*250 mm, 5 μm; solvent: A = water + trifluoroacetic acid (0.1%), B = acetonitrile; gradient: 20-50%; 20 mL / min) to give N-((1R,3S)-3-((6-chloro-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)-1-(2-(methylsulfonyl)ethyl)-1H-pyrazole-4-carboxamide trifluoroacetate (36.0 mg, 9.9% yield).

[0162] 1H NMR (400MHz, CD3OD): δ8.54(d,1H),8.18(s,1H),7.97-7.93(m,2H),7.89-7.83(m,1H),7.17(s,1H),4.66(t,2H),4.11-3.98(m ,2H),3.72(t,2H),2.82(s,3H),2.42-2.32(m,1H),2.15-2.07(m,1H),2.03-1.95(m,2H),1.68-1.51(m,3H),1.49-1.39(m,1H).

[0163] LC-MS, M / Z(ESI):544.2[M+H] +

[0164] Example 2: Preparation of 3-chloro-N-((1R,3S))-3-((6-chloro-2-trifluoromethyl))quinolin-4-yl)amino)cyclohexyl)-1-(2-(methylsulfonyl)ethyl)-1H-pyrazole-4-carboxamide (Compound 5)

[0165] The synthetic route of compound 5 is as follows:

[0166] Step 1: Synthesis of ethyl 3-chloro-1-(2-(methylsulfonyl)ethyl)-1H-pyrazole-4-carboxylate

[0167] At room temperature, ethyl 3-chloro-1H-pyrazole-4-carboxylate (0.500 g, 2.86 mmol) was dissolved in N,N-dimethylformamide (5 mL), followed by the addition of potassium carbonate (0.792 mg, 5.73 mmol), sodium iodide (4.00 mg, 0.286 mmol), and 2-methylsulfonylethyl bromide (589 mg, 3.15 mmol). The reaction mixture was stirred at 25° C. for 3 hours. After completion of the reaction, the reaction mixture was filtered, the filtrate was dried, and the crude product was purified by column chromatography (PE:EA (V / V) = 10:1 to 1:1) to obtain ethyl 3-chloro-1-(2-(methylsulfonyl)ethyl)-1H-pyrazole-4-carboxylate (0.600 g, 74.8% yield).

[0168] LC / MS (ESI) (m / z): 281.0 (M+H) + .

[0169] Step 2: Synthesis of 3-chloro-1-(2-(methylsulfonyl)ethyl)-1H-pyrazole-4-carboxylic acid

[0170] At room temperature, ethyl 3-chloro-1-(2-(methylsulfonyl)ethyl)-1H-pyrazole-4-carboxylate (0.300 g, 1.07 mmol) was dissolved in tetrahydrofuran (2 mL) and methanol (1 mL). A solution of lithium hydroxide (128 mg, 5.34 mmol) in water (1 mL) was then slowly added. After the addition was complete, the mixture was heated to 25°C and stirred for 12 hours. After completion of the reaction, the pH of the reaction solution was adjusted to 3 using 4N HCl solution in an ice bath. Solid precipitated and was filtered. The filter cake was collected and dried to yield 3-chloro-1-(2-(methylsulfonyl)ethyl)-1H-pyrazole-4-carboxylic acid (200 mg, 73.8% yield).

[0171] LC / MS (ESI) (m / z): 253.0 (M+H) +

[0172] Step 3: Synthesis of 3-chloro-N-((1R,3S))-3-((6-chloro-2-trifluoromethyl))quinolin-4-yl)amino)cyclohexyl)-1-(2-(methylsulfonyl)ethyl)-1H-pyrazole-4-carboxamide

[0173] At room temperature, (1S,3R)-N 1 -(6-Chloro-2-(trifluoromethyl))-quinolin-4-yl)cyclohexane-1,3-diamine (50.0 mg, 0.150 mmol) and 3-chloro-1-(2-(methylsulfonyl)ethyl)-1H-pyrazole-4-carboxylic acid (40.0 mg, 0.160 mmol) were dissolved in N,N-dimethylformamide (0.5 mL), followed by the addition of diisopropylethylamine (56.0 mg, 0.440 mmol) and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (86.0 mg, 0.230 mmol), and the reaction solution was stirred at room temperature for 2 hours. After the reaction was completed, the reaction solution was filtered, the filtrate was spin-dried, and the crude product was purified by preparative liquid chromatography (chromatographic column: YMC-Triart Prep C18 7μm 30mm×40cm; solvent: A=0.1% ammonia water, B=acetonitrile; gradient: 30%-87%, 9 minutes) to give 3-chloro-N-((1R,3S))-3-((6-chloro-2-trifluoromethyl))quinolin-4-yl)amino)cyclohexyl)-1-(2-(methylsulfonyl)ethyl)-1H-pyrazole-4-carboxamide (Compound 5) (18.0 mg, yield 20.7%).

[0174] LC / MS (ESI) (m / z): 578.0 (M+H) + ;

[0175] 1H NMR(400MHz, CDCl3)δ8.07(s,1H),8.01(d,1H),7.66(s,1H),7.63(d,1H),6.76(s,1H),6.41(d,1H),4.96(d,1H),4.56(t,2H),4.14(d, 1H),3.71(d,1H),3.60(t,2H),2.74(s,3H),2.59(d,1H),2.25(d,1H),2.17(d,1H),2.01–1.96(m,1H),1.61(d,1H),1.36–1.28(m,3H).

[0176] Example 3: Preparation of 3-chloro-N-((1R,3S))-3-((6-chloro-2-trifluoromethyl))quinolin-4-yl)amino)cyclohexyl)-1-((methylsulfonyl)methyl)-1H-pyrazole-4-carboxamide (Compound 7)

[0177] The synthetic route of compound 7 is as follows:

[0178] Step 1: Synthesis of 3-chloro-1-((methylsulfonyl)methyl)-1H-pyrazole-4-carboxylic acid (7-2)

[0179] Compound 3-chloro-1-((methylsulfonyl)methyl)-1H-pyrazole-4-carboxylic acid ethyl ester (7-1) (80 mg, 0.30 mmol) was dissolved in a mixture of methanol (2 mL) and water (0.8 mL). Sodium hydroxide (96 mg, 2.40 mmol) was then added, and the reaction mixture was stirred at room temperature for 2 h. After TLC monitoring indicated that the reaction was complete, stirring was stopped, and the methanol was removed by distillation under reduced pressure. The residue was diluted with water (5 mL), and the pH was adjusted to 2 with dilute hydrochloric acid (2 mol / L). The mixture was then extracted with ethyl acetate (15 mL x 3). The organic phase was collected and dried over anhydrous sodium sulfate. The organic phase was then concentrated by distillation under reduced pressure to obtain compound 3-chloro-1-((methylsulfonyl)methyl)-1H-pyrazole-4-carboxylic acid (70 mg, 97.79% yield). The crude product was used directly in the next reaction.

[0180] Step 2: Synthesis of 3-chloro-N-((1R,3S))-3-((6-chloro-2-trifluoromethyl))quinolin-4-yl)amino)cyclohexyl)-1-((methylsulfonyl)methyl)-1H-pyrazole-4-carboxamide (Compound 7)

[0181] Compound 3-chloro-1-((methylsulfonyl)methyl)-1H-pyrazole-4-carboxylic acid (7-2) (70 mg, 0.29 mmol) was dissolved in anhydrous DMF (2 mL). The reaction solution was stirred at 0°C for 10 min, and then DIPEA (114 mg, 0.88 mmol) and HATU (223 mg, 0.59 mmol) were added. After stirring at 0°C for 30 min, (1S,3R)-N 1 -(6-chloro-2-(trifluoromethyl))-quinolin-4-yl)cyclohexane-1,3-diamine (Intermediate A1) (111 mg, 0.32 mmol), the resulting reaction solution was slowly warmed to room temperature and stirred for 6 h. After TLC monitoring showed that the reaction of the starting materials was complete, stirring was stopped and water (5 mL) was added to the reaction solution to quench the reaction. The reaction solution was then extracted with ethyl acetate (15 mL × 3). The organic phase was collected and dried over anhydrous sodium sulfate. The organic phase was concentrated by distillation under reduced pressure, and the residue was separated and purified by silica gel column chromatography (DCM:MeOH (V / V) = 100:3) to obtain compound 3-chloro-N-((1R,3S))-3-((6-chloro-2-trifluoromethyl))quinolin-4-yl)amino)cyclohexyl)-1-((methylsulfonyl)methyl)-1H-pyrazole-4-carboxamide (Compound 7) (47 mg, 28.39% yield).

[0182] LC-MS, M / Z (ESI): 564.0 (M+1)

[0183] 1 H NMR(400MHz,DMSO-d6)δ8.59(d,1H),8.36(s,1H),8.11(d,1H),7.89(d,1H),7.74(dd,1H),7.48(d,1H),6. 94(s,1H),5.79(s,2H),4.04–3.80(m,2H),3.07(s,3H),2.16(d,1H),2.03–1.72(m,3H),1.62–1.15(m,4H).

[0184] Example 4: Preparation of 2-(4-(((1R,3S)-3-((6-chloro-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)carbamoyl)-1H-pyrazol-1-yl)ethyl dimethylcarbamate (Compound 8)

[0185] The synthetic route of compound 8 is as follows:

[0186] Step 1: Synthesis of ethyl 1-(2-hydroxyethyl)-1H-pyrazole-4-carboxylate

[0187] To a solution of ethyl 2-formyl-3-oxopropanoate (10.0 g, 69.4 mmol) and 4A molecular sieves (6.00 g, 69.4 mmol) in ethanol (40 mL) was added dropwise a solution of 2-diazepin-1-ol (5.28 g, 69.4 mmol) in ethanol (40 mL) at 0°C, and the resulting mixture was stirred at 25°C for 13 hours. The reaction solution was filtered through celite, and the filtrate was concentrated in vacuo. The crude product was separated and purified using a reverse-phase column (water:acetonitrile (V / V) = 100:0-10:1, gradient elution) to obtain ethyl 1-(2-hydroxyethyl)-1H-pyrazole-4-carboxylate (6.50 g, 50.9% yield).

[0188] LC-MS, M / Z(ESI):185.2[M+H] +

[0189] Step 2: Ethyl 1-(2-((dimethylcarbamoyl)oxy)ethyl)-1H-pyrazole-4-carboxylate

[0190] To a solution of ethyl 1-(2-hydroxyethyl)-1H-pyrazole-4-carboxylate (1.50 g, 8.14 mmol) in tetrahydrofuran (20 mL) was added N,N'-carbonyldiimidazole (1.98 g, 12.22 mmol). The resulting mixture was then stirred at 25 ° C for 1 hour under nitrogen protection, and then dimethylamine (0.73 g, 16.4 mmol) was slowly added to the reaction solution via syringe. The resulting mixture was stirred at 25 ° C for 12 hours. The reaction mixture was concentrated in vacuo, added to dichloromethane (20 mL) and hydrochloric acid (10 mL, 1N aqueous solution), separated, and the aqueous phase was extracted with dichloromethane (20 mL × 2). The organic layers were combined, washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, and concentrated to obtain a crude product. The product was separated and purified by silica gel column (methanol:dichloromethane (V / V) = 100:1-100:5, gradient elution) to give ethyl 1-(2-((dimethylcarbamoyl)oxy)ethyl)-1H-pyrazole-4-carboxylate (900 mg, yield 43.3%).

[0191] LC-MS, M / Z(ESI):256.2[M+H] +

[0192] Step 3: 1-(2-((dimethylcarbamoyl)oxy)ethyl)-1H-pyrazole-4-carboxylic acid

[0193] To a solution of ethyl 1-(2-((dimethylcarbamoyl)oxy)ethyl)-1H-pyrazole-4-carboxylate (450 mg, 1.76 mmol) in tetrahydrofuran (5 mL) and water (5 mL) was added lithium hydroxide (148 mg, 3.53 mmol), and the reaction solution was stirred at 25° C. for 3 hours. The reaction solution was concentrated under reduced pressure and then purified by reverse phase column separation (water:acetonitrile (v / v) = 100:0-100:1) to give 1-(2-((dimethylcarbamoyl)oxy)ethyl)-1H-pyrazole-4-carboxylic acid (240 mg, 59.9% yield).

[0194] LC-MS, M / Z(ESI):228.2[M+H] +

[0195] Step 4: 2-(4-(((1R,3S)-3-((6-chloro-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)carbamoyl)-1H-pyrazol-1-yl)ethyl dimethylcarbamate (Compound 8)

[0196] To (1S,3R)-N 1 To a solution of 1-(6-chloro-2-(trifluoromethyl)quinolin-4-yl)cyclohexane-1,3-diamine (Intermediate A1) (50.0 mg, 0.15 mmol) in N,N-dimethylformamide (1.5 mL) were added 1-(2-((dimethylcarbamoyl)oxy)ethyl)-1H-pyrazole-4-carboxylic acid (50.0 mg, 0.22 mmol), N,N-diisopropylethylamine (0.07 mL, 0.44 mmol) and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (111 mg, 0.29 mmol). The resulting mixture was then stirred at 25° C. under nitrogen for 12 hours. The reaction mixture was purified by preparative high performance liquid chromatography (chromatographic column: Kinetex EVO C18, 21.2*250mm, 5μm; 0.05% NH3.H2O-ACN; 37~67%; 20mL / min) to give 2-(4-(((1R,3S)-3-((6-chloro-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)carbamoyl)-1H-pyrazol-1-yl)ethyl dimethylcarbamate (Compound 8) (22.2 mg, 27.6% yield).

[0197] LC-MS, M / Z(ESI):553.2[M+H] +

[0198] 1H NMR (400MHz, CD3OD): δ8.38(d,1H),8.12(s,1H),7.98-7.88(m,2H),7.69(dd,1H),6.90(s,1H),4.45-4.37(m,4H),4.15-4.03(m,1H), 3.89-3.71(m,1H),2.89-2.80(m,6H),2.43-2.35(m,1H),2.16-2.09(m,1H),2.05-1.94(m,2H),1.67-1.57(m,1H),1.56-1.38(m,3H).

[0199] Example 5: Preparation of N-(1R,3S)-3-((6-chloro-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)-4-(dimethylphosphoryl)benzamide (Compound 32)

[0200] The synthetic route of compound 32 is as follows:

[0201] Step 1: Synthesis of methyl 4-(dimethylphosphoryl)benzoate

[0202] Methyl 4-iodobenzoate (2.00 g, 7.63 mmol) was dissolved in dimethyl sulfoxide (20 mL). Potassium phosphate (4.86 g, 22.9 mmol), 1,3-bis(diphenylphosphino)propane (0.63 g, 1.53 mmol), and palladium acetate (0.17 g, 0.76 mmol) were added sequentially under nitrogen. The reaction was stirred at 25°C for 10 minutes. A solution of dimethylphosphine oxide (0.89 g, 11.4 mmol) in dimethyl sulfoxide (1 mL) was then added. The reaction mixture was reacted at 130°C under nitrogen for 1 hour. The reaction mixture was cooled to room temperature, diluted with water (100 mL), and then extracted with ethyl acetate (100 mL*3). The organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was purified by reverse phase chromatography (water:acetonitrile (V / V) = 100:1 to 2:1, gradient elution) to give methyl 4-(dimethylphosphoryl)benzoate (1.50 g, yield 87.9%).

[0203] LC-MS, M / Z(ESI):213.2[M+H] + .

[0204] Step 2: Synthesis of 4-(dimethylphosphoryl)benzoic acid

[0205] Methyl 4-(dimethylphosphoryl)benzoate (1.00 g, 4.48 mmol) was dissolved in water (2 mL) and methanol (20 mL), and lithium hydroxide (3.76 g, 89.5 mmol) was added. The reaction solution was reacted at 25°C for 1 hour. The reaction solution was adjusted to pH = 2 with hydrochloric acid (2 M), then washed with ethyl acetate (10 mL × 3), and the aqueous phase was concentrated to obtain a crude product. The crude product was purified by reverse phase chromatography (water:acetonitrile (V / V) = 100:1-10:3, gradient elution) to obtain 4-(dimethylphosphoryl)benzoic acid (750 mg, 82.6% yield).

[0206] LC-MS, M / Z(ESI):199.4[M+H] +

[0207] Step 3: N-(1R,3S)-3-((6-chloro-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)-4-(dimethylphosphoryl)benzamide (Compound 32)

[0208] 4-(Dimethylphosphoryl)benzoic acid (104 mg, 0.52 mmol) was dissolved in dimethyl sulfoxide (2 mL), and (1S,3R)-N 1 -(6-chloro-2-(trifluoromethyl)quinolin-4-yl)cyclohexane-1,3-diamine (Intermediate A1) (150 mg, 0.44 mmol), N,N-diisopropylethylamine (0.36 mL, 2.18 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (251 mg, 1.31 mmol), and 1-hydroxybenzotriazole (174 mg, 1.31 mmol). The reaction solution was stirred at 25°C under nitrogen for 18 hours. The reaction solution was diluted with saturated brine (10 mL) and then extracted with ethyl acetate (10 mL × 3). The organic layers were combined, dried over anhydrous sodium sulfate, and concentrated to obtain a crude product. The product was separated and purified by silica gel column (dichloromethane:methanol (V / V) = 100:1-10:1, gradient elution) to obtain N-(1R,3S)-3-((6-chloro-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)-4-(dimethylphosphoryl)benzamide (Compound 32) (25.0 mg, 10.8% yield).

[0209] 1H NMR (400MHz, CD3OD): δ8.38(d,1H),7.97(dd,2H),7.94-7.80(m,3H),7.69(dd,1H),6.92(s,1H),4.24-4.05(m,1H),3.91-3.7 4(m,1H),2.42(d,1H),2.21-2.10(m,1H),2.10-2.02(m,1H),2.02-1.93(m,1H),1.82(s,3H),1.79(s,3H),1.69-1.43(m,4H).

[0210] LC-MS, M / Z(ESI):523.8[M+H] +

[0211] Example 6: Preparation of N-((1R,3S)-3-((6-chloro-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)-4-(dimethylphosphoryl)benzamide (Compound 33)

[0212] The synthetic route of compound 33 is as follows:

[0213] Step 1: Synthesis of methyl 4-bromo-3-((tert-butoxycarbonyl)amino)benzoate

[0214] To a solution of methyl 3-amino-4-bromobenzoate (5.00 g, 21.7 mmol) in dichloromethane (50 mL) were added triethylamine (9.06 mL, 65.2 mmol), 4-dimethylaminopyridine (0.27 g, 2.17 mmol), and di-tert-butyl dicarbonate (7.49 mL, 32.6 mmol) in sequence. The reaction mixture was stirred at 25°C for 12 hours. The reaction mixture was quenched with saturated brine (50 mL) and extracted with dichloromethane (50 mL x 3). The organic layers were combined, dried over anhydrous sodium sulfate, and concentrated to afford the crude product, which was purified on a silica gel column (petroleum ether:ethyl acetate (v / v) = 1:0-10:1, gradient elution) to afford methyl 4-bromo-3-((tert-butoxycarbonyl)amino)benzoate (2.01 g, 27.9% yield).

[0215] Step 2: Synthesis of methyl 3-amino-4-(dimethylphosphoryl)benzoate

[0216] To a solution of methyl 4-bromo-3-((tert-butoxycarbonyl)amino)benzoate (1.90 g, 5.75 mmol) in dimethyl sulfoxide (15 mL) were added palladium acetate (129 mg, 0.58 mmol), 1,3-bis(diphenylphosphino)propane (475 mg, 1.15 mmol), and potassium phosphate (3.66 mg, 17.3 mmol) in sequence. The reaction mixture was stirred at 25°C under nitrogen for 5 minutes. A solution of dimethylphosphine oxide (665 mg, 8.63 mmol) in dimethyl sulfoxide (5 mL) was then added, and the mixture was stirred at 130°C for 1 hour. The reaction mixture was cooled to room temperature, filtered, and the filtrate was purified by reverse phase chromatography (water:acetonitrile (v / v) = 100:0 to 5:3, gradient elution) to afford methyl 3-amino-4-(dimethylphosphino)benzoate (800 mg, 61.2% yield).

[0217] LC-MS, M / Z(ESI):228.0[M+H] +

[0218] Step 3: Synthesis of methyl 4-(dimethylphosphoryl)-3-(methylamino)benzoate

[0219] To a solution of methyl 3-amino-4-(dimethylphosphoryl)benzoate (500 mg, 2.20 mmol) in methanol (5 mL) were added acetic acid (0.01 mL, 0.22 mmol), paraformaldehyde (330 mg, 11.0 mmol), and sodium cyanoborohydride (415 mg, 6.60 mmol) in sequence. The reaction was stirred at 50°C for 12 hours. The reaction solution was quenched with water (5 mL) and concentrated under reduced pressure to afford the crude product. The crude product was purified by silica gel column chromatography (dichloromethane:methanol (v / v) = 100:1 to 10:1, gradient elution) to afford methyl 4-(dimethylphosphoryl)-3-(methylamino)benzoate (450 mg, 74.6% yield).

[0220] LC-MS, M / Z(ESI):242.2[M+H] +

[0221] Step 4: 4-(dimethylphosphoryl)-3-(methylamino)benzoic acid

[0222] To a solution of methyl 4-(dimethylphosphoryl)-3-(methylamino)benzoate (250 mg, 1.04 mmol) in methanol (5 mL) and water (1 mL) was added lithium hydroxide (87.0 mg, 2.07 mmol), and the reaction mixture was stirred at 25°C for 2 hours. The reaction mixture was spin-dried to dryness, and the crude product was purified by reverse-phase column chromatography (water:acetonitrile (V / V) = 100:1-10:1, gradient elution) to afford 4-(dimethylphosphoryl)-3-(methylamino)benzoic acid (200 mg, 84.9% yield).

[0223] LC-MS, M / Z(ESI):228.0[M+H] +

[0224] Step 5: N-((1R,3S)-3-((6-chloro-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)-4-(dimethylphosphoryl)benzamide

[0225] To a solution of 4-(dimethylphosphoryl)-3-(methylamino)benzoic acid (150 mg, 0.66 mmol) in dimethyl sulfoxide (5 mL) were added 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (253 mg, 1.32 mmol), 1-hydroxybenzotriazole (178 mg, 1.32 mmol), (1S,3R)-N 1 -(6-chloro-2-(trifluoromethyl)quinolin-4-yl)cyclohexane-1,3-diamine (Intermediate A1) (227 mg, 0.66 mmol) and N,N-diisopropylethylamine (0.55 mL, 3.30 mmol) were added, and the reaction mixture was stirred at 25°C for 2 hours. The reaction mixture was diluted with saturated brine (50 mL) and then extracted with ethyl acetate (50 mL x 3). The organic layers were combined, dried over anhydrous sodium sulfate, and concentrated to obtain a crude product. The crude product was purified by HPLC (column: Xtimate C18, 21.2*250 mm, 5 μm; 0.05% NH3.H2O-ACN; 45%-75%; 60 mL / min) to give N-((1R,3S)-3-((6-chloro-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)-4-(dimethylphosphoryl)benzamide (compound 33) (74.0 mg, 20.3% yield).

[0226] 1H NMR (400MHz, DMSO-d6): δ8.61(d,1H),8.30(d,1H),7.89(d,1H),7.74(dd,1H),7.50 (d,1H),7.46–7.39(m,1H),7.38-7.28(m,1H),7.01(d,1H),6.97–6.90(m,2H),4.10 –3.97(m,1H),3.97–3.82(m,1H),2.78(d,3H),2.20–2.13(m,1H),2.01–1.94(m,1H) ,1.92–1.79(m,2H),1.70(s,3H),1.67(s,3H),1.60–1.49(m,2H),1.45–1.29(m,2H).

[0227] LC-MS, M / Z (ESI): 553.2 (M+H) +

[0228] Example 7: Preparation of N-((1R,3S)-3-((6-chloro-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)-3-(dimethylphosphoryl)benzamide (Compound 36)

[0229] The synthetic route of compound 36 is as follows:

[0230] Step 1: Synthesis of methyl 3-(dimethylphosphoryl)benzoate

[0231] Methyl 3-iodobenzoate (3.00 g, 11.4 mmol) was dissolved in dimethyl sulfoxide (30 mL), and potassium phosphate (4.86 g, 22.9 mmol), 1,3-bis(diphenylphosphino)propane (0.63 g, 1.53 mmol), and palladium acetate (129 mg, 0.57 mmol) were added in sequence under nitrogen protection. The reaction solution was stirred at 25 ° C. for 10 minutes under nitrogen protection, and then a solution of dimethyl phosphine oxide (1.34 g, 17.2 mmol) in dimethyl sulfoxide (3 mL) was added. The mixture was stirred at 130 ° C. for 2 hours under N2 protection. The reaction mixture was cooled to room temperature, diluted with water (100 mL), and then extracted with ethyl acetate (50 mL×3). The aqueous layer was retained and concentrated to give a crude product, which was separated and purified using a reverse-phase silica gel column (C18, acetonitrile:water (V / V) = 0:100-10:90, gradient elution) to give compound 3-(dimethylphosphoryl)benzoic acid methyl ester (2.20 g, yield 88.7%).

[0232] LC-MS, M / Z(ESI):213.4[M+H] + .

[0233] Step 2: Synthesis of 3-(dimethylphosphoryl)benzoic acid

[0234] Methyl 3-(dimethylphosphoryl)benzoate (1.00 g, 4.71 mmol) was dissolved in water (5 mL) and methanol (5 mL), and lithium hydroxide (1.98 g, 47.1 mmol) was added. The reaction solution was reacted at 25°C for 1 hour, then adjusted to pH 4 with dilute hydrochloric acid (2 M), washed with ethyl acetate (10 mL x 3), and the aqueous layer was retained and concentrated to obtain a crude product. The crude product was purified by reverse phase chromatography on a C18 column (acetonitrile:water (V / V) = 0:100-10:90, gradient elution) to obtain compound 3-(dimethylphosphoryl)benzoic acid (600 mg, 64.3% yield).

[0235] LC-MS, M / Z(ESI):199.2[M+H] + .

[0236] Step 3: Synthesis of N-((1R,3S)-3-((6-chloro-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)-3-(dimethylphosphoryl)benzamide (Compound 36)

[0237] 3-(Dimethylphosphoryl)benzoic acid (170 mg, 0.86 mmol) was dissolved in dimethyl sulfoxide (2 mL), and (1S,3R)-N 1 -(6-chloro-2-(trifluoromethyl)quinolin-4-yl)cyclohexane-1,3-diamine (Intermediate A1) (295 mg, 0.86 mmol), N,N-diisopropylethylamine (846 mg, 6.55 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (493 mg, 2.57 mmol), 1-hydroxybenzotriazole (348 mg, 2.57 mmol), the reaction solution was stirred at 25 ° C for 18 hours under N2 protection. The resulting mixture was diluted with saturated brine (10 mL), then extracted with ethyl acetate (10 mL × 3), the organic layers were combined, dried over anhydrous sodium sulfate, filtered and concentrated to obtain a crude product. The crude product was purified by silica gel column chromatography (methanol:dichloromethane (V / V) = 0:100-10:90, gradient elution) to give N-((1R,3S)-3-((6-chloro-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)-3-(dimethylphosphoryl)benzamide (Compound 36) (193 mg, 42.5% yield).

[0238] 1H NMR (400MHz, CD3OD): δ8.39(d,1H),8.26–8.19(m,1H),8.08–8.02(m,1H),8.00–7.89(m,2H),7.72–7.63(m,2H),6.93(s,1H),4.20–4.09 (m,1H),3.90–3.79(m,1H),2.51–2.40(m,1H),2.21–2.12(m,1H),2.11–2.05(m,1H),2.03–1.95(m,1H),1.83(d,6H),1.69–1.42(m,4H).

[0239] LC-MS, M / Z(ESI):524.2[M+H] + .

[0240] Example 8: Preparation of N-((1R,3S)-3-((6-chloro-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)-3-(dimethylphosphoryl)benzamide (Compound 38)

[0241] The synthetic route of compound 38 is as follows:

[0242] Step 1: Synthesis of ethyl 3-iodo-1-(2,2,2-trifluoroethyl)-1H-pyrazole-4-carboxylate

[0243] To ethyl 3-iodo-1H-pyrazole-4-carboxylate (2.00 g, 7.52 mmol) in tetrahydrofuran (20 mL) was added sodium hydride (0.45 g, 11.3 mmol, 60% Wt) at 0°C. The mixture was stirred at 0°C for 30 minutes, followed by the addition of 2,2,2-trifluoroethyl trifluoromethanesulfonate (2.62 g, 11.3 mmol). The reaction mixture was stirred at 25°C under N2 protection for 30 minutes. LC-MS confirmed the reaction was complete. The reaction mixture was diluted with water (50 mL) and extracted with ethyl acetate (50 mL x 3). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to yield the crude product. The crude product was separated and purified by silica gel column (petroleum ether:ethyl acetate (V / V) = 100:0-80:20, gradient elution) to give ethyl 3-iodo-1-(2,2,2-trifluoroethyl)-1H-pyrazole-4-carboxylate (370 mg, yield 12.7%).

[0244] LC-MS, M / Z(ESI):349.2[M+H] + .

[0245] Step 2: Synthesis of ethyl 3-(dimethylphosphoryl)-1-(2,2,2-trifluoroethyl)-1H-pyrazole-4-carboxylate

[0246] Methyl 3-iodo-1-(2,2,2-trifluoroethyl)pyrazole-4-carboxylate (320 mg, 0.92 mmol) was dissolved in dimethyl sulfoxide (2 mL). Potassium phosphate (585 mg, 2.76 mmol), 1,3-bis(diphenylphosphino)propane (75.8 mg, 0.18 mmol), and palladium acetate (20.6 mg, 0.09 mmol) were added sequentially under nitrogen. The reaction solution was stirred at 25°C for 10 minutes, followed by the addition of a solution of dimethylphosphine oxide (108 mg, 1.38 mmol) in dimethyl sulfoxide (2 mL). The resulting mixture was stirred at 130°C for 2 hours under nitrogen. LC-MS confirmed the reaction was complete. The reaction mixture was diluted with water (10 mL) and extracted with ethyl acetate (10 mL × 3). The aqueous layer was retained and concentrated to obtain the crude product. The crude product was separated and purified using a C18 column (water:acetonitrile (V / V) = 100:0-90:10, gradient elution) to give compound 3-(dimethylphosphoryl)-1-(2,2,2-trifluoroethyl)-1H-pyrazole-4-carboxylic acid ethyl ester (100 mg, yield 35.2%).

[0247] LC-MS, M / Z(ESI):299.2[M+H] + .

[0248] Step 3: Synthesis of 3-(dimethylphosphoryl)-1-(2,2,2-trifluoroethyl)-1H-pyrazole-4-carboxylic acid

[0249] Ethyl 3-(dimethylphosphoryl)-1-(2,2,2-trifluoroethyl)-1H-pyrazole-4-carboxylate (100 mg, 0.34 mmol) was dissolved in water (1 mL) and methanol (5 mL), and lithium hydroxide (141 mg, 3.35 mmol) was added. The resulting mixture was stirred at 25°C for 1 hour. LC-MS analysis confirmed the reaction was complete. The reaction solution was adjusted to pH 4 with hydrochloric acid (2 mol / L) and extracted with ethyl acetate (10 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to afford 3-(dimethylphosphoryl)-1-(2,2,2-trifluoroethyl)-1H-pyrazole-4-carboxylic acid (70.0 mg, 77.3% yield). The crude product was used directly in the next step without further purification.

[0250] LC-MS, M / Z(ESI):271.1[M+H] + .

[0251] Step 4: Synthesis of N-((1R,3S)-3-((6-chloro-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)-3-(dimethylphosphoryl)-1-(2,2,2-trifluoroethyl)-1H-pyrazole-4-carboxamide

[0252] 3-(Dimethylphosphoryl)-1-(2,2,2-trifluoroethyl)-1H-pyrazole-4-carboxylic acid (60.0 mg, 0.22 mmol) was dissolved in DMSO (2 mL), and (1S,3R)-N 1 -(6-chloro-2-(trifluoromethyl)quinolin-4-yl)cyclohexane-1,3-diamine (Intermediate A1) (76.4 mg, 0.22 mmol), N,N-diisopropylethylamine (144 mg, 1.11 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (144 mg, 1.11 mmol), 1-hydroxybenzotriazole (90.0 mg, 0.67 mmol), and the resulting mixture was stirred at 25 ° C for 18 hours under nitrogen protection. The reaction was completed by LC-MS detection. The reaction solution was diluted with saturated brine (20 mL), then extracted with ethyl acetate (10 mL×3). The organic layers were combined, dried over anhydrous sodium sulfate, filtered and concentrated to give a crude product, which was purified by preparative liquid chromatography (Xtimate C18, 21.2*250 mm, 5 μm; 0.1% TFA-ACN; 38-68; 20 mL / min) to give compound N-((1R,3S)-3-((6-chloro-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)-3-(dimethylphosphoryl)-1-(2,2,2-trifluoroethyl)-1H-pyrazole-4-carboxamide (Compound 38) (12.6 mg, yield 9.5%).

[0253] LC-MS, M / Z(ESI):596.2[M+H] + .

[0254] 1 H NMR (400MHz, CD3OD): δ8.52–8.32(m,2H),7.93(d,1H),7.76–7.63(m,1H),6.92(s,1H),5.13(q,2H),4.15–3.97(m,1H ),3.94–3.73(m,1H),2.52–2.36(m,1H),2.17–2.06(m,2H),2.02–1.87(m,7H),1.72–1.59(m,1H),1.59–1.36(m,3H).

[0255] Example 9: Preparation of N-((1R,3S)-3-((6-chloro-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)-3-(dimethylphosphoryl)benzamide (Compound 40)

[0256] The synthetic route of compound 40 is as follows:

[0257] Step 1: Synthesis of ethyl 3-iodo-1-methyl-1H-pyrazole-4-carboxylate

[0258] Ethyl 3-iodo-1H-pyrazole-4-carboxylate (2.00 g, 7.518 mmol) was dissolved in anhydrous N,N-dimethylformamide (20 mL). Cesium carbonate (2.45 g, 7.52 mmol) and iodomethane (1.28 g, 9.02 mmol) were added. The reaction mixture was stirred at 25°C under N2 protection for 1 hour. LC-MS confirmed the formation of the desired product. The reaction mixture was diluted with water (100 mL) and extracted with ethyl acetate (50 mL x 3). The organic phases were combined, washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, and filtered and concentrated to obtain the crude product. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate (v / v) = 100:0-80:20, gradient elution) to obtain ethyl 3-iodo-1-methyl-1H-pyrazole-4-carboxylate (900 mg, 41.9% yield).

[0259] LC-MS, M / Z(ESI):281.1[M+H] + .

[0260] Step 2: Synthesis of ethyl 3-(dimethylphosphoryl)-1-methyl-1H-pyrazole-4-carboxylate

[0261] Ethyl 3-iodo-1-methyl-1H-pyrazole-4-carboxylate (600 mg, 2.14 mmol) was dissolved in dimethyl sulfoxide (5 mL). Potassium phosphate (1.36 g, 6.43 mmol), 1,3-bis(diphenylphosphino)propane (139 mg, 0.43 mmol), and palladium acetate (48.5 mg, 0.21 mmol) were added sequentially under nitrogen. The reaction solution was stirred at 25°C for 10 minutes, followed by the addition of a solution of dimethylphosphine oxide (251 mg, 3.21 mmol) in dimethyl sulfoxide (1 mL). The resulting mixture was stirred at 135°C for 1 hour under nitrogen protection. LC-MS confirmed the reaction was complete. The reaction mixture was diluted with water (10 mL) and extracted with ethyl acetate (10 mL × 3). The aqueous layer was retained and concentrated to obtain the crude product. The crude product was separated and purified using a reverse phase C18 column (water:acetonitrile (V / V) = 100:0-90:10, gradient elution) to give compound 3-(dimethylphosphoryl)-1-methyl-1H-pyrazole-4-carboxylic acid ethyl ester (250 mg, yield 50.7%).

[0262] LC-MS, M / Z(ESI):231.2[M+H] + .

[0263] Step 3: Synthesis of 3-(dimethylphosphoryl)-1-methyl-1H-pyrazole-4-carboxylic acid

[0264] To a solution of ethyl 3-(dimethylphosphoryl)-1-methyl-1H-pyrazole-4-carboxylate (200 mg, 0.87 mmol) in water (10 mL) and methanol (2 mL) was added lithium hydroxide (365 mg, 8.69 mmol), and the resulting mixture was stirred at 25° C. for 1 hour. LC-MS confirmed the completion of the reaction, and the methanol was removed by concentration under reduced pressure. The resulting reaction mixture was adjusted to pH 5 with hydrochloric acid (2 mol / L), resulting in the precipitation of a solid. The filter cake was collected by filtration and purified using a reversed phase C18 column (water:acetonitrile (V / V) = 100:0-60:40, gradient elution) to afford 3-(dimethylphosphoryl)-1-methyl-1H-pyrazole-4-carboxylic acid (120 mg, 68.3% yield).

[0265] LC-MS, M / Z (ESI): 203.0 [MH] + .

[0266] Step 4: Synthesis of N-((1R,3S)-3-((6-chloro-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)-3-(dimethylphosphoryl)benzamide (Compound 40)

[0267] 3-(Dimethylphosphoryl)-1-methyl-1H-pyrazole-4-carboxylic acid (100 mg, 0.49 mmol) was dissolved in DMSO (2 mL), and (1S,3R)-N 1 -(6-chloro-2-(trifluoromethyl)quinolin-4-yl)cyclohexyl-1,3-diamine (Intermediate A1) (170 mg, 0.49 mmol), N,N-diisopropylethylamine (320 mg, 2.47 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (284 mg, 1.48 mmol), 1-hydroxybenzotriazole (201 mg, 1.48 mmol), the resulting mixture was stirred at 25 ° C. under N2 protection for 18 hours. LC-MS detection showed that the reaction was complete. The reaction solution was diluted with saturated brine (10 mL), then extracted with ethyl acetate (10 mL × 3). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product. The crude product was separated and purified on a normal phase silica gel column (petroleum ether:ethyl acetate (V / V) = 100:1-0:100, gradient elution) to give compound N-((1R,3S)-3-((6-chloro-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)-3-(dimethylphosphoryl)benzamide (Compound 40) (47.4 mg, yield 17.9%).

[0268] 1 H NMR (400MHz, CD3OD): δ8.38(d,1H),8.20(s,1H),7.92(d,1H),7.70(dd,J=9.2,2.4Hz 1H),6.91(s,1H),4.06–4.00(m,1H),3.98(s,3H),3.85–3.78(m,1H),2.44–2.38(m,1H),2.15–2.05(m,2H ),1.99–1.94(m,1H),1.90(d,3H),1.86(d,3H),1.68–1.59(m,1H),1.53–1.43(m,2H),1.38–1.34(m,1H).

[0269] LC-MS, M / Z(ESI):528.2[M+H] + .

[0270] Example 10: Preparation of (4-((1R,3S)-3-((6-chloro-2-trifluoromethyl))quinolin-4-yl)amino)cyclohexyl)carbamoyl)-1-methyl-1H-pyrazol-3-yl)methyl dimethylcarbamate (Compound 41)

[0271] The synthetic route of compound 41 is as follows:

[0272] Step 1: Synthesis of methyl 3-formyl-1-methyl-1H-pyrazole-4-carboxylate

[0273] At room temperature, 4-bromo-1-methyl-pyrazole-3-carbaldehyde (2.50 g, 13.2 mmol) and triethylamine (4.00 g, 39.68 mmol) were dissolved in methanol (25 mL). [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (484 mg, 0.662 mmol) was then added. After complete addition, the reaction mixture was stirred at 60°C under a carbon monoxide atmosphere (15 psi) for 12 hours. After completion of the reaction, the reaction mixture was cooled to room temperature, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by column chromatography (PE:EA (V / V) = 10:1 to 1:1) to afford methyl 3-formyl-1-methyl-1H-pyrazole-4-carboxylate (1.10 g, 49.5% yield).

[0274] LC / MS (ESI) (m / z): 169.0 (M+H) + .

[0275] Step 2: Synthesis of methyl 3-(hydroxymethyl)-1-methyl-1H-pyrazole-4-carboxylate

[0276] At room temperature, 3-formyl-1-methyl-1H-pyrazole-4-carboxylic acid methyl ester (1.10 g, 6.54 mmol) was dissolved in methanol (20 mL), and then sodium borohydride (272 mg, 7.20 mmol) was added in portions under an ice bath. The reaction solution was stirred under an ice bath for 2 hours. After the reaction was completed, water (20 mL) was slowly added to quench the reaction, and the mixture was extracted with ethyl acetate (3 × 20 mL). The organic phase was collected and then washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was purified by column chromatography (PE:EA (V / V) = 10:1t to 0:1) to obtain compound 3-(hydroxymethyl)-1-methyl-1H-pyrazole-4-carboxylic acid methyl ester (800 mg, yield 71.9%).

[0277] LC / MS (ESI) (m / z): 171.0 (M+H) + .

[0278] Step 3: Synthesis of methyl 3-((dimethylaminoformyloxy)methyl)-1-methyl-1H-pyrazole-4-carboxylate

[0279] Sodium hydride (85.0 mg, 2.12 mmol) was dissolved in tetrahydrofuran (3 mL) at room temperature, and then methyl 3-(hydroxymethyl)-1-methyl-1H-pyrazole-4-carboxylate (300 mg, 1.76 mmol) was slowly added at 0°C. The reaction solution was stirred at room temperature for 1 hour, and then dimethylcarbamoyl chloride (284 mg, 2.64 mmol) was added dropwise. After the addition, the temperature was raised to 80°C and stirred for 1 hour. The reaction solution was cooled to room temperature and quenched with saturated ammonium chloride (2 mL) in an ice bath. The mixture was then extracted with dichloromethane (5 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain crude methyl 3-((dimethylcarbamoyloxy)methyl)-1-methyl-1H-pyrazole-4-carboxylate (424 mg, 1.76 mmol, 100% yield), which was used directly in the next reaction.

[0280] LC / MS (ESI) (m / z): 242.1 (M+H) + .

[0281] Step 4: Synthesis of 3-((dimethylaminoformyloxy)methyl)-1-methyl-1H-pyrazole-4-carboxylic acid

[0282] To a solution of methyl 3-((dimethylcarbamoyloxy)methyl)-1-methyl-1H-pyrazole-4-carboxylate (424 mg, 1.76 mmol, 100% yield) in tetrahydrofuran (3 mL) was added dropwise a solution of lithium hydroxide (127 mg, 5.29 mmol) in water (3 mL) at 0°C, and the reaction solution was stirred at room temperature for 12 hours. After completion of the reaction, the reaction solution was adjusted to pH 3 with 1N HCl solution in an ice bath, extracted with ethyl acetate (3×10 mL), and the organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to afford 3-((dimethylcarbamoyloxy)methyl)-1-methyl-1H-pyrazole-4-carboxylic acid (300 mg, 75% yield).

[0283] LC / MS (ESI) (m / z): 228.0 (M+H) +

[0284] Step 5: Synthesis of (4-((1R,3S)-3-((6-chloro-2-trifluoromethyl))quinolin-4-yl)amino)cyclohexyl)carbamoyl)-1-methyl-1H-pyrazol-3-yl)methyl dimethylcarbamate (Compound 41)

[0285] At room temperature, (1S,3R)-N 1-(6-Chloro-2-(trifluoromethyl)quinolin-4-yl)cyclohexyl-1,3-diamine (Intermediate A1) (45.0 mg, 0.130 mmol) and 3-((dimethylaminoformyloxy)methyl)-1-methyl-1H-pyrazole-4-carboxylic acid (30.0 mg, 0.130 mmol) were dissolved in N,N-dimethylformamide (0.5 mL), followed by the addition of diisopropylethylamine (51.6 mg, 0.400 mmol) and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (49.0 mg, 0.130 mmol), and the reaction mixture was stirred at 25 ° C for 2 hours. After the reaction was completed, the reaction solution was filtered and the filtrate was dried by rotary evaporation. The crude product was purified by preparative liquid chromatography (chromatographic column: YMC-Triart Prep C18 7 μm 30 mm × 40 cm; solvent: A = 0.1% ammonia water, B = acetonitrile; gradient: 30%-70%, 9 minutes) to give (4-((1R,3S)-3-((6-chloro-2-trifluoromethyl))quinolin-4-yl)amino)cyclohexyl)carbamoyl)-1-methyl-1H-pyrazol-3-yl)methyl dimethylcarbamate (Compound 41) (14.9 mg, 20.7% yield).

[0286] LC / MS (ESI) (m / z): 553.10 (M+H) + ;

[0287] 1 H NMR(400MHz,DMSO-d6)δ8.54(d,1H),8.07(s,1H),7.83(t,2H),7.69(dd,1H),7.43(d,1H),6.88(s,1H),5.10(s,2H),3.89(d,1H),3 .81(d,1H),3.77(s,3H),2.72(s,6H),2.09(d,1H),1.91(d,1H),1.79(dd,2H),1.48(d,1H),1.39(d,1H),1.32(d,1H),1.19(d,1H).

[0288] Example 11: N-((1R,3S)-3-((6-chloro-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)-3-(dimethyl(oxo)-λ 6 Preparation of 1-(2-hydroxy-2-methylpropyl)-1H-pyrazole-4-carboxamide (Compound 42)

[0289] The synthetic route of compound 42 is as follows:

[0290] Step 1: Synthesis of ethyl 1-(2-hydroxy-2-methylpropyl)-3-iodo-1H-pyrazole-4-carboxylate

[0291] To a solution of ethyl 3-iodo-1H-pyrazole-4-carboxylate (2.00 g, 7.52 mmol) in N,N-dimethylformamide (20 mL) was added cesium carbonate (2.45 g, 7.52 mmol) and iodomethane (1.28 g, 9.02 mmol), and the resulting mixture was stirred at 25 ° C for 1 hour under N2 protection. LC-MS detection showed that the reaction was complete, and the reaction solution was diluted with water (100 mL), then extracted with ethyl acetate (100 mL × 3), and the organic layers were combined, washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, and filtered and concentrated to obtain the crude product. The crude product was separated and purified by a normal silica gel column (petroleum ether: ethyl acetate (V / V) = 100:0-80:20, gradient elution) to obtain compound 1-(2-hydroxy-2-methylpropyl)-3-iodo-1H-pyrazole-4-carboxylic acid ethyl ester (900 mg, yield 41.9%).

[0292] Step 2: 3-((dimethyl(oxo)-λ 6 Synthesis of ethyl 1-(2-hydroxy-2-methylpropyl)-1H-pyrazole-4-carboxylate

[0293] To a solution of ethyl 1-(2-hydroxy-2-methylpropyl)-3-iodo-1H-pyrazole-4-carboxylate (700 mg, 2.07 mmol) in 1,4-dioxane (10 mL) were added dimethylsulfenyl imide (289 mg, 3.11 mmol), cesium carbonate (1.01 mg, 3.11 mmol), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (180 mg, 0.31 mmol), and tris(dibenzylideneacetone)dipalladium (94.8 mg, 0.10 mmol) in sequence. The resulting mixture was stirred at 110°C for 18 hours. LC-MS confirmed the completion of the reaction. The reaction solution was diluted with water (50 mL) and extracted with ethyl acetate (50 mL x 3). The combined organic layers were washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to yield the crude product. The crude product was separated and purified by normal silica gel column (dichloromethane: methanol (V / V) = 100:0-90:10, gradient elution) to obtain compound 3-((dimethyl(oxo)-λ 6 -aminosulfonyl imide)-1-(2-hydroxy-2-methylpropyl)-1H-pyrazole-4-carboxylic acid ethyl ester (480 mg, yield 68.8%).

[0294] LC-MS, M / Z(ESI):304.2[M+H] + .

[0295] Step 3: 3-((dimethyl(oxo)-λ 6 Synthesis of (sulfapyridinyl)amino)-1-(2-hydroxy-2-methylpropyl)-1H-pyrazole-4-carboxylic acid

[0296] To 3-((dimethyl(oxy)-λ 6 To a solution of ethyl 1-(2-hydroxy-2-methylpropyl)-1H-pyrazole-4-carboxylate (200 mg, 0.66 mmol) in water (5 mL) and methanol (1 mL) was added lithium hydroxide (55.3 mg, 1.32 mmol), and the resulting mixture was stirred at 20°C for 18 hours. LC-MS analysis confirmed the completion of the reaction. The reaction solution was concentrated under reduced pressure to remove methanol, and the resulting mixture was adjusted to pH 5 with hydrochloric acid (2 mol / L). The filter cake was collected after filtration and dried to yield compound 3-((dimethyl(oxo)-1,6-sulfapyridinyl)amino)-1-(2-hydroxy-2-methylpropyl)-1H-pyrazole-4-carboxylic acid (180 mg, 89.3% yield). The crude product was used directly in the next step without further purification.

[0297] LC-MS, M / Z(ESI):276.2[MH] - .

[0298] Step 4: N-((1R,3S)-3-((6-chloro-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)-3-(dimethyl(oxo)-λ 6 Synthesis of 1-(2-hydroxy-2-methylpropyl)-1H-pyrazole-4-carboxamide

[0299] 3-((dimethyl(oxo)-λ 6 Sulfapyridinyl)amino)-1-(2-hydroxy-2-methylpropyl)-1H-pyrazole-4-carboxylic acid (100 mg, 0.36 mmol) was dissolved in dimethyl sulfoxide (2 mL), and (1S,3R)-N 1-(6-chloro-2-(trifluoromethyl)quinolin-4-yl)cyclohexyl-1,3-diamine (Intermediate A1) (125 mg, 0.36 mmol), N,N-diisopropylethylamine (46.9 mg, 0.36 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (69.6 mg, 0.36 mmol), 1-hydroxybenzotriazole (49.1 mg, 0.36 mmol), the resulting mixture was stirred at 60 ° C for 18 hours under N2 protection. LC-MS detection showed that the reaction was complete. The reaction solution was diluted with saturated brine (20 mL), then extracted with ethyl acetate (20 mL × 3), and the organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product. The crude product was separated and purified by silica gel column (petroleum ether: ethyl acetate (V / V) = 100:0-50:50, gradient elution) to obtain the compound N-((1R,3S)-3-((6-chloro-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)-3-(dimethyl(oxo)-λ 6 -aminosulfonamide)-1-(2-hydroxy-2-methylpropyl)-1H-pyrazole-4-carboxamide (Compound 42) (109 mg, yield 48.6%).

[0300] 1 H NMR (400MHz, DMSO-d6): δ8.58(d,1H),7.93–7.84(m,2H),7.82(s,1H),7.74(dd,1H),7.47(d,1H),6.93(s,1H),4.65(s,1H),4.04–3.72(m,4H) ,3.38(s,6H),2.29–2.20(m,1H),2.01–1.93(m,2H),1.87–1.76(m,1H) ,1.62–1.48(m,1H),1.44–1.32(m,2H),1.22–1.12(m,1H),1.06(s,6H).

[0301] LC-MS, M / Z(ESI):601.2[M+H] + .

[0302] Example 12: Preparation of 2-(3-chloro-4-((1R,3S)-3-((6-chloro-2-trifluoromethyl))quinolin-4-yl)amino)cyclohexyl)carbamoyl)-1H-pyrazol-1-yl)ethyl dimethylcarbamate (Compound 43)

[0303] The synthetic route of compound 43 is as follows:

[0304] Step 1: Synthesis of ethyl 3-chloro-1-(2-hydroxyethyl)-1H-pyrazole-4-carboxylate

[0305] 3-Chloro-1H-pyrazole-4-carboxylic acid ethyl ester (0.500 g, 2.86 mmol) and potassium carbonate (0.792 g, 5.73 mmol) were added to acetonitrile (15 mL), followed by the addition of 2-bromoethanol (394 mg, 3.15 mmol). The reaction solution was heated to 60° C. and stirred for 12 hours. After completion of the reaction, the reaction solution was filtered and the filtrate was concentrated. The crude product was purified by column chromatography (PE:EA (V / V) = 10:1 to 1:1) to obtain the compound 3-chloro-1-(2-hydroxyethyl)-1H-pyrazole-4-carboxylic acid ethyl ester (0.6 g, 96.0% yield).

[0306] LC / MS (ESI) (m / z): 219.0 (M+H) + .

[0307] Step 2: Synthesis of ethyl 3-chloro-1-(2-((dimethylcarbamoyl)oxy)ethyl)-1H-pyrazole-4-carboxylate

[0308] Sodium hydride (66.0 mg, 1.65 mmol, 60%) was dissolved in tetrahydrofuran (3 mL). 3-Chloro-1-(2-hydroxyethyl)-1H-pyrazole-4-carboxylic acid ethyl ester (300 mg, 1.37 mmol) was slowly added under ice-bath. The reaction solution was warmed to room temperature and stirred for 1 hour. Dimethylaminoformyl chloride (221 mg, 2.06 mmol) was then added dropwise under ice-bath. After the addition was complete, the reaction solution was warmed to room temperature and stirred for 12 hours. The reaction solution was cooled to room temperature and quenched with saturated ammonium chloride (2 mL) under ice-bath. The mixture was then extracted with dichloromethane (5 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to give the crude compound 3-chloro-1-(2-((dimethylaminoformyl)oxy)ethyl)-1H-pyrazole-4-carboxylic acid ethyl ester (397 mg, 1.37 mmol, 100% yield), which was used directly in the next reaction.

[0309] LC / MS (ESI) (m / z): 290.1 ​​(M+H) + .

[0310] Step 3: Synthesis of 3-chloro-1-(2-((dimethylcarbamoyl)oxy)ethyl)-1H-pyrazole-4-carboxylic acid

[0311] To a solution of ethyl 3-chloro-1-(2-((dimethylcarbamoyl)oxy)ethyl)-1H-pyrazole-4-carboxylate (397 mg, 1.37 mmol) in tetrahydrofuran (3 mL) was added dropwise a solution of lithium hydroxide (164 mg, 6.83 mmol) in water (3 mL) at 0°C. The reaction solution was stirred at room temperature for 12 hours. After completion of the reaction, the pH of the reaction solution was adjusted to 3 with 4N HCl solution in an ice bath, then extracted with ethyl acetate (3 x 10 mL). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to afford the crude compound 3-chloro-1-(2-((dimethylcarbamoyl)oxy)ethyl)-1H-pyrazole-4-carboxylic acid (300 mg, 83% yield).

[0312] LC / MS (ESI) (m / z): 262.0 (M+H) +

[0313] Step 4: Synthesis of 2-(3-chloro-4-((1R,3S)-3-((6-chloro-2-trifluoromethyl))quinolin-4-yl)amino)cyclohexyl)carbamoyl)-1H-pyrazol-1-yl)ethyl dimethylcarbamate (Compound 43)

[0314] (1S,3R)-N 1 -(6-Chloro-2-(trifluoromethyl))-quinolin-4-yl)cyclohexane-1,3-diamine (Intermediate A1) (50.0 mg, 0.150 mmol) and 3-chloro-1-(2-((dimethylaminoformyl)oxy)ethyl)-1H-pyrazole-4-carboxylic acid (57.0 mg, 0.220 mmol) were dissolved in N,N-dimethylformamide (0.5 mL), followed by the addition of diisopropylethylamine (56.0 mg, 0.440 mmol) and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (84.0 mg, 0.220 mmol), and the reaction mixture was stirred at 25 ° C for 2 hours. After the reaction was completed, the reaction solution was filtered and the filtrate was dried. The crude product was purified by preparative high performance liquid chromatography (chromatographic column: YMC-Triart Prep C18 7μm 30mm×40cm; solvent: A=0.1% ammonia water, B=acetonitrile; gradient: 30%-82%, 9 minutes) to obtain compound 2-(3-chloro-4-((1R,3S)-3-((6-chloro-2-trifluoromethyl))quinolin-4-yl)amino)cyclohexyl)carbamoyl)-1H-pyrazol-1-yl)ethyl dimethylcarbamate (Compound 43) (24.4 mg, yield 27.7%).

[0315] LC / MS (ESI) (m / z): 587.10 (M+H) + ;

[0316] 1 H NMR(400MHz,DMSO-d6)δ8.55(d,1H),8.23(s,1H),7.85(d,1H),7.76(d,1H),7.69(dd,1H),7.43(d,1H),6.89(s,1H),4.27(d,2H),4.23(d,2H), 3.89(d,1H),3.81(s,1H),2.72(d,6H),2.11(d,1H),1.93(s,1H),1.84( d,1H),1.76(d,1H),1.49(d,1H),1.42(d,1H),1.33(s,1H),1.24(d,1H).

[0317] Example 13: Preparation of 3-chloro-N-((1R,3S)-3-((6-chloro-2-trifluoromethyl))quinolin-4-yl)amino)cyclohexyl)-1-(1-(methylsulfonyl)ethyl)-1H-pyrazole-4-carboxamide (Compound 44)

[0318] The synthetic route of compound 44 is as follows:

[0319] Step 1: Synthesis of ethyl 3-chloro-1-((methylthio)methyl)-1H-pyrazole-4-carboxylate

[0320] At room temperature, ethyl 3-chloro-1H-pyrazole-4-carboxylate (2.00 g, 11.46 mmol) was dissolved in anhydrous tetrahydrofuran (20 mL) and stirred in an ice bath at 0°C for 10 min. Sodium hydride (504 mg, 12.60 mmol) was then added portionwise, followed by the slow addition of chloromethyl methyl sulfide (1.33 g, 13.75 mmol). The reaction solution was slowly warmed to room temperature and stirred for 30 min. After completion of the reaction, stirring was stopped and the reaction solution was poured into water (30 mL), followed by extraction with ethyl acetate (30 mL × 3). The organic phase was collected and dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was separated and purified by silica gel column chromatography (petroleum ether:ethyl acetate (V / V) = 10:1) to obtain ethyl 3-chloro-1-((methylthio)methyl)-1H-pyrazole-4-carboxylate (930 mg, 34.59% yield).

[0321] LC-MS, M / Z(ESI):235.3[M+H] +

[0322] 1H NMR (400MHz, DMSO-d6) δ8.50(s,1H),5.26(s,2H),4.23(q,2H),2.14(s,3H),1.27(t,3H).

[0323] Step 2: Synthesis of ethyl 3-chloro-1-((methylsulfonyl)methyl)-1H-pyrazole-4-carboxylate

[0324] Ethyl 3-chloro-1-((methylthio)methyl)-1H-pyrazole-4-carboxylate (550 mg, 2.34 mmol) was dissolved in anhydrous dichloromethane (11 mL) at room temperature and stirred at 0°C for 10 min. m-Chloroperbenzoic acid (1.21 g, 7.03 mmol) was then added. The reaction mixture was slowly warmed to room temperature and stirred for 2 h. After thin-layer chromatography indicated complete reaction, stirring was stopped and the reaction mixture was poured into a saturated aqueous sodium thiosulfate solution (30 mL). The mixture was then extracted with ethyl acetate (30 mL x 3). The organic phase was collected and washed with a saturated aqueous sodium carbonate solution (40 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated to afford ethyl 3-chloro-1-((methylsulfonyl)methyl)-1H-pyrazole-4-carboxylate (615 mg, 98.40% yield). The crude product was used directly in the next reaction.

[0325] LC-MS, M / Z(ESI):267.0[M+H] +

[0326] Step 3: Synthesis of ethyl 3-chloro-1-(1-(methylsulfonyl)ethyl)-1H-pyrazole-4-carboxylate

[0327] Ethyl 3-chloro-1-((methylsulfonyl)methyl)-1H-pyrazole-4-carboxylate (200 mg, 0.75 mmol) was dissolved in anhydrous DMF (2 mL) at room temperature, followed by the addition of cesium carbonate (488.67 mg, 1.50 mmol). The reaction mixture was stirred for 10 minutes, and then iodomethane (212.88 mg, 1.50 mmol) was slowly added dropwise. Stirring was continued at room temperature for 12 hours. After TLC monitoring indicated complete reaction, stirring was stopped and the reaction was quenched with water (5 mL). The reaction was then extracted with ethyl acetate (15 mL x 3). The organic phase was collected, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silica gel column chromatography (DCM:MeOH (v / v) = 100:2) to afford ethyl 3-chloro-1-(1-(methylsulfonyl)ethyl)-1H-pyrazole-4-carboxylate (139 mg, 66.03% yield).

[0328] LC-MS, M / Z(ESI):281.0[M+H] +

[0329] Step 4: Synthesis of compound 3-chloro-1-(1-(methylsulfonyl)ethyl)-1H-pyrazole-4-carboxylic acid

[0330] Ethyl 3-chloro-1-(1-(methylsulfonyl)ethyl)-1H-pyrazole-4-carboxylate (130 mg, 0.46 mmol) was dissolved in a mixture of methanol (2 mL) and water (0.8 mL) at room temperature. Sodium hydroxide (148 mg, 3.70 mmol) was then added, and the reaction mixture was stirred at room temperature for 2 h. After TLC monitoring indicated complete reaction, stirring was stopped and the methanol was removed by distillation under reduced pressure. The residue was diluted with water (5 mL) and then adjusted to pH 2 with dilute hydrochloric acid (2 mol / L). The mixture was extracted with ethyl acetate (15 mL x 3). The organic phase was collected and dried over anhydrous sodium sulfate. The organic phase was then concentrated by distillation under reduced pressure to afford 3-chloro-1-(1-(methylsulfonyl)ethyl)-1H-pyrazole-4-carboxylic acid (110 mg, 94.01% yield). The crude product was used directly in the next reaction.

[0331] LC-MS, M / Z(ESI):252.8[M+H] +

[0332] Step 5: Synthesis of 3-chloro-N-((1R,3S)-3-((6-chloro-2-trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)-1-(1-(methylsulfonyl)ethyl)-1H-pyrazole-4-carboxamide (Compound 44)

[0333] At room temperature, compound 3-chloro-1-(1-(methylsulfonyl)ethyl)-1H-pyrazole-4-carboxylic acid (1119E) (50 mg, 0.20 mmol) was dissolved in anhydrous DMF (1 mL) and stirred at 0°C for 10 min. DIPEA (102 mg, 0.79 mmol) and HATU (151 mg, 0.40 mmol) were then added. Stirring was continued at 0°C for 30 min. Then, (1S,3R)-N 1-(6-chloro-2-(trifluoromethyl))-quinolin-4-yl)cyclohexane-1,3-diamine (Intermediate A1) (68 mg, 0.20 mmol) was added, and the reaction solution was slowly warmed to room temperature and stirred for 6 h. After TLC monitoring showed that the reaction of the starting material was complete, stirring was stopped and water (5 mL) was added to quench the reaction. The mixture was then extracted with ethyl acetate (15 mL × 3). The organic phase was collected and dried over anhydrous sodium sulfate. The organic phase was concentrated by distillation under reduced pressure, and the residue was separated and purified by silica gel column chromatography (DCM:MeOH (V / V) = 100:3) to obtain compound 3-chloro-N-((1R,3S)-3-((6-chloro-2-trifluoromethyl))quinolin-4-yl)amino)cyclohexyl)-1-(1-(methylsulfonyl)ethyl)-1H-pyrazole-4-carboxamide (Compound 44) ​​(55 mg, 48.05% yield).

[0334] LC-MS, M / Z(ESI):578.2[M+H] +

[0335] 1 H NMR(400MHz,DMSO-d6)δ8.59(d,1H),8.47(d,1H),8.04(dd,1H),7.89(d,1H),7.74(dd,1H),7.48(d,1H),6.94 (s,1H),5.91(q,1H),3.92(dd,2H),2.99(d,3H),2.17(d,1H),1.93(dd,2H),1.82(dd,3H),1.61–1.19(m,5H).

[0336] Example 14: Preparation of N-((1R,3S))-3-((6-chloro-2-trifluoromethyl))(quinolin-4-yl)amino)cyclohexyl)-3-((methylsulfonyl)methyl)benzamide (Compound 45)

[0337] The synthetic route of compound 45 is as follows:

[0338] At room temperature, (1S,3R)-N 1-(6-chloro-2-(trifluoromethyl))-quinolin-4-yl)cyclohexane-1,3-diamine (80.0 mg, 0.230 mmol), 3-((methylsulfonyl)methyl)benzoic acid (50.0 mg, 0.230 mmol) and N,N-diisopropylethylamine (45.0 mg, 0.350 mmol) were dissolved in N,N-dimethylformamide (1.0 mL), and then 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (133 mg, 0.350 mmol) was added. After the addition was complete, the reaction solution was stirred at room temperature for 12 hours. After completion of the reaction, the reaction solution was concentrated under reduced pressure, and the crude product was purified by preparative high performance liquid chromatography (chromatographic column: YMC-Triart Prep C18 7 μm 30 mm × 40 cm; solvent: A = 0.1% ammonia water, B = acetonitrile; gradient: 30%-72%, 9 minutes) to give compound N-((1R,3S))-3-((6-chloro-2-trifluoromethyl))(quinolin-4-yl)amino)cyclohexyl)-3-((methylsulfonyl)methyl)benzamide (Compound 44) ​​(62.0 mg, 49.9% yield).

[0339] LC / MS (ESI) (m / z): 540.10 (M+H) + ;

[0340] 1 H NMR(400MHz,DMSO-d6)δ8.59(d,1H),8.37(d,1H),7.85(dd,3H),7.72(dd,1H),7.49(dt,3H),6.93(s,1H),4.52(s,2H) ,4.08–4.00(m,1H),3.92–3.84(m,1H),2.90(d,3H),2.17(d,1H),1.97–1.78(m,3H),1.53(dd,2H),1.41–1.29(m,2H).

[0341] Example 15: Preparation of 3-chloro-N-((1R,3S))-3-((6-chloro-2-trifluoromethyl))quinolin-4-yl)amino)cyclohexyl)-1-(2-(methylsulfonyl)propan-2-yl)-1H-pyrazole-4-carboxamide (Compound 46)

[0342] The synthetic route of compound 46 is as follows:

[0343] Step 1: Synthesis of ethyl 3-chloro-1-(1-(methylsulfonyl)ethyl)-1H-pyrazole-4-carboxylate

[0344] At room temperature, ethyl 3-chloro-1-(1-(methylsulfonyl)ethyl)-1H-pyrazole-4-carboxylate (44-4) (0.138 g, 0.490 mmol) was dissolved in N,N-dimethylformamide (2 mL) and tetrahydrofuran (2 mL), and then sodium hydroxide (60%, 40.0 mg, 0.980 mmol) was slowly added under ice bath. The reaction solution was stirred under ice bath for 20 minutes, and then iodomethane (140 mg, 0.980 mmol) was slowly added. After the addition, the reaction solution was heated to 25°C and stirred for 12 hours. After the reaction was completed, saturated ammonium chloride solution (2 mL) was added dropwise under ice bath to quench the reaction, and the mixture was extracted with ethyl acetate (5 mL*3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered and concentrated to obtain compound 3-chloro-1-(2-(methylsulfonyl)propan-2-yl)-1H-pyrazole-4-carboxylic acid ethyl ester (46-2) (0.140 g, yield 97.1%).

[0345] LC / MS (ESI) (m / z): 295.0 (M+H) + .

[0346] Step 2: Synthesis of 3-chloro-1-(1-(methylsulfonyl)ethyl)-1H-pyrazole-4-carboxylic acid

[0347] At room temperature, ethyl 3-chloro-1-(2-(methylsulfonyl)propan-2-yl)-1H-pyrazole-4-carboxylate (46-2) (0.140 g, 0.476 mmol) was dissolved in tetrahydrofuran (2 mL) and methanol (1 mL). A solution of lithium hydroxide (100 mg, 2.38 mmol) in water (1 mL) was then slowly added. The reaction mixture was stirred at 25°C for 12 hours. After completion of the reaction, the pH of the reaction mixture was adjusted to 3 with 4N HCl solution under ice-cooling, and then extracted with ethyl acetate (5 mL*3). The organic phases were combined, washed with saturated brine (5 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to afford 3-chloro-1-(1-(methylsulfonyl)ethyl)-1H-pyrazole-4-carboxylic acid (46-3) (126 mg, 100% yield).

[0348] LC / MS (ESI) (m / z): 267.0 (M+H) +

[0349] Step 3: Synthesis of 3-chloro-N-((1R,3S))-3-((6-chloro-2-trifluoromethyl))quinolin-4-yl)amino)cyclohexyl)-1-(2-(methylsulfonyl)propan-2-yl)-1H-pyrazole-4-carboxamide (Compound 46)

[0350] At room temperature, (1S,3R)-N 1-(6-Chloro-2-(trifluoromethyl))-quinolin-4-yl)cyclohexane-1,3-diamine (Intermediate A1) (81.0 mg, 0.240 mmol) and 3-chloro-1-(1-(methylsulfonyl)ethyl)-1H-pyrazole-4-carboxylic acid (46-3) (63.0 mg, 0.240 mmol) were dissolved in N,N-dimethylformamide (1.0 mL), and then diisopropylethylamine (61.0 mg, 0.470 mmol) and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (110 mg, 0.280 mmol) were added, and the reaction solution was stirred at 25 ° C for 12 hours. After the reaction was completed, the reaction solution was filtered and the filtrate was concentrated. The crude product was purified by preparative high performance liquid chromatography (chromatographic column: YMC-Actus Triart C18ExRS-5μm 100*30mm; solvent: A=0.1% FA, B=acetonitrile; gradient: 30-85%, 9 min; 85-98% 0.1 min; 85-98% 2 min.) to give 3-chloro-N-((1R,3S))-3-((6-chloro-2-trifluoromethyl))quinolin-4-yl)amino)cyclohexyl)-1-(2-(methylsulfonyl)propan-2-yl)-1H-pyrazole-4-carboxamide (Compound 46) (9.70 mg, yield 6.8%).

[0351] LC / MS (ESI) (m / z): 592.0 (M+H) + ;

[0352] 1 H NMR(400MHz,DMSO-d6)δ8.61(s,1H),8.56(s,1H),7.96(d,1H),7.86(d,1H),7.71(d,1H),7.44(d,1H),6.9 1(s,1H),3.93(d,1H),3.86(s,1H),2.89(s,3H),2.15(d,1H),2.01–1.83(m,8H),1.79(d,1H),1.43(m,4H).

[0353] Example 16: Preparation of 3-chloro-N-((1R,3S))-3-((6-chloro-2-trifluoromethyl))quinolin-4-yl)amino)cyclohexyl)-1-((S-methylsulfonylimino)methyl)-1H-pyrazole-4-carboxamide (Compound 47)

[0354] The synthetic route of compound 47 is as follows:

[0355] Step 1: Synthesis of ethyl 3-chloro-1-((S-methylsulfonylimino)methyl)-1H-pyrazole-4-carboxylate

[0356] At room temperature, ethyl 3-chloro-1-((methylthio)methyl)-1H-pyrazole-4-carboxylate (44-2) (400 mg, 1.70 mmol) was dissolved in anhydrous methanol (15 mL). Iodophenyldiacetic acid (1.70 g, 5.10 mmol) and ammonium carbonate (490 mg, 5.10 mmol) were then added. After the addition was complete, the mixture was stirred at room temperature for 4 hours. After TLC monitoring indicated complete reaction of the starting materials, stirring was stopped, water (50 mL) was added for dilution, and the mixture was extracted with ethyl acetate (30 mL x 3). The organic phase was collected, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silica gel column chromatography (PE:EA = 100:10 (V / V)) to obtain ethyl 3-chloro-1-((S-methylsulfonylimino)methyl)-1H-pyrazole-4-carboxylate (47-1) (376 mg, 83.03% yield).

[0357] LC-MS, M / Z(ESI):266.0[M+H] + .

[0358] Step 2: Synthesis of 3-chloro-1-((S-methylsulfonylimino)methyl)-1H-pyrazole-4-carboxylic acid (47-2)

[0359] At room temperature, ethyl 3-chloro-1-((S-methylsulfonylimino)methyl)-1H-pyrazole-4-carboxylate (47-1) (200 mg, 0.75 mmol) was dissolved in a mixture of tetrahydrofuran / methanol / water (2 mL / 1 mL / 1 mL). Lithium hydroxide monohydrate (253 mg, 6.02 mmol) was then slowly added under ice-cooling. After the addition was complete, the reaction solution was stirred at room temperature for 2 hours. After TLC monitoring indicated complete reaction of the starting materials, stirring was stopped and the reaction solution was diluted with water (10 mL). The pH was adjusted to 4 with dilute hydrochloric acid (1 mol / L), followed by extraction with ethyl acetate (30 mL x 3). The organic phase was collected, dried over anhydrous sodium sulfate, filtered, and concentrated to yield 3-chloro-1-((S-methylsulfonylimino)methyl)-1H-pyrazole-4-carboxylic acid (47-2) (150 mg, 83.86% yield).

[0360] LC-MS, M / Z(ESI):238.0[M+H] + .

[0361] Step 3: Synthesis of 3-chloro-N-((1R,3S))-3-((6-chloro-2-trifluoromethyl))quinolin-4-yl)amino)cyclohexyl)-1-((S-methylsulfonylimino)methyl)-1H-pyrazole-4-carboxamide (Compound 47)

[0362] At room temperature, 3-chloro-1-((S-methylsulfonylimino)methyl)-1H-pyrazole-4-carboxylic acid (47-2) (50 mg, 0.21 mmol) was dissolved in anhydrous N,N-dimethylformamide (1 mL) and stirred in an ice bath for 10 minutes. Then, N,N-diisopropylethylamine (109 mg, 0.84 mmol) and N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)uronium hexafluorophosphate (160 mg, 0.42 mmol) were added. The reaction solution was stirred in an ice bath for 30 minutes, and then (1S,3R)-N 1 After addition, the mixture was slowly heated to room temperature and stirred for 6 hours. After TLC monitoring indicated complete reaction of the starting materials, stirring was stopped and the mixture was diluted with water (5 mL). The mixture was then extracted with ethyl acetate (5 mL × 3). The organic phase was collected and dried over anhydrous sodium sulfate. The organic phase was concentrated by distillation under reduced pressure, and the residue was purified by silica gel column chromatography (DCM:MeOH = 100:5 (V / V)) to obtain 3-chloro-N-((1R,3S))-3-((6-chloro-2-trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)-1-((S-methylsulfonylimino)methyl)-1H-pyrazole-4-carboxamide (Compound 47) (23 mg, 19.40% yield).

[0363] LC-MS, M / Z(ESI):563.0[M+H] + .

[0364] 1 H NMR (400MHz, DMSO-d6): δ8.59(d,1H),8.33(s,1H),8.03(d,1H),7.89(d,1H),7.74(dd,1H),7.48(d,1H),6.94(s,1H),5.64 -5.39(m,2H),4.02(d,1H),3.99-3.81(m,1H),2.92(d,3H),2.16(d,1H),2.03-1.76(m,3H),1.61-1.22(m,5H).

[0365] Example 17: Preparation of 3-chloro-N-((1R,3S)-3-((6-chloro-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)-1-((N,S-dimethylsulfonylimino)methyl)-1H-pyrazole-4-carboxamide (Compound 51)

[0366] The synthetic route of compound 51 is as follows:

[0367] Step 1: 3-Chloro-1-((N,S-dimethylsulfonylimino)methyl)-1H-pyrazole-4-carboxylic acid ethyl ester

[0368] At room temperature, ethyl 3-chloro-1-((S-methylsulfonylimino)methyl)-1H-pyrazole-4-carboxylate (47-1) (50.0 mg, 0.190 mmol) was dissolved in N,N-dimethylformamide (1 mL), and then cesium carbonate (61.0 mg, 0.190 mmol) and iodomethane (32.0 mg, 0.230 mmol) were added at room temperature. After the addition, the mixture was stirred at 25°C for 12 hours. After the reaction was completed, the reaction solution was diluted with ethyl acetate (20 mL), washed with water (10 mL) and saturated brine (10 mL) in sequence, and the organic phase was concentrated under reduced pressure to obtain the compound ethyl 3-chloro-1-((N,S-dimethylsulfonylimino)methyl)-1H-pyrazole-4-carboxylate (51-1) (53.0 mg, 100% yield). LC / MS (ESI) (m / z): 280.0 (M+H) + .

[0369] Step 2: 3-chloro-1-((N,S-dimethylsulfonylimino)methyl)-1H-pyrazole-4-carboxylic acid

[0370] At room temperature, ethyl 3-chloro-1-((N,S-dimethylsulfonylimino)methyl)-1H-pyrazole-4-carboxylate (51-1) (53.0 mg, 0.190 mmol) was dissolved in a mixture of tetrahydrofuran (0.4 mL) / methanol (0.2 mL) / water (0.2 mL). Lithium hydroxide monohydrate (40.0 mg, 0.950 mmol) was then added at room temperature. The reaction mixture was stirred at 25°C for 12 hours. After completion of the reaction, the pH was adjusted to 3 with dilute hydrochloric acid (1 mol / L) under ice-cooling, followed by extraction with ethyl acetate (20 mL), followed by washing with water (5 mL) and saturated brine (5 mL). The organic phase was concentrated under reduced pressure to afford 3-chloro-1-((N,S-dimethylsulfonylimino)methyl)-1H-pyrazole-4-carboxylic acid (51-2) (48.0 mg, 0.190 mmol, 100% yield). LC / MS (ESI) (m / z): 252.0 (M+H) +

[0371] Step 3: 3-Chloro-N-((1R,3S)-3-((6-chloro-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)-1-((N,S-dimethylsulfonylimino)methyl)-1H-pyrazole-4-carboxamide (Compound 51)

[0372] At room temperature, 3-chloro-1-((N,S-dimethylsulfonylimino)methyl)-1H-pyrazole-4-carboxylic acid (51-2) (48.0 mg, 0.190 mmol) was dissolved in N,N-dimethylformamide (1.0 mL), and then diisopropylethylamine (0.100 mL, 0.570 mmol), (7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (87.0 mg, 0.228 mmol) and (1R,3S)-3-{[6-chloro-2-(trifluoromethyl)quinolin-4-yl]amino}cyclohexan-1-amine (66.0 mg, 0.190 mmol) were added in sequence. The reaction solution was stirred at 25°C for 12 hours. After the reaction, the reaction solution was diluted with ethyl acetate (20 mL), washed with water (10 mL) and saturated brine (10 mL) in that order, and the organic phase was concentrated under reduced pressure. The crude product was purified by preparative high performance liquid chromatography (chromatographic column: YMC-Triart Prep C18 7 μm 30 mm × 40 cm; solvent: A = 0.1% ammonia water, B = acetonitrile; gradient: 25%-76%, 9 minutes) to give 3-chloro-N-((1R,3S)-3-((6-chloro-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)-1-((N,S-dimethylsulfonylimino)methyl)-1H-pyrazole-4-carboxamide (Compound 51) (4.90 mg, 4.5% yield).

[0373] LC / MS (ESI) (m / z): 577.0 (M+H) + ;

[0374] 1 H NMR (400MHz, DMSO-d6): δ8.56(s,1H),8.29(s,1H),8.01(d,1H),7.86(d,1H),7.70(d,1H),7.45(d,1H),6.90(s,1H),5. 65(d,2H),3.92(s,1H),3.83(s,1H),2.92(s,3H),2.61(s,3H),2.12(s,1H),1.96(d,1H),1.85(s,1H),1.76(s,1H),1.52 -1.48(m,3H),1.42-1.39(m,1H).

[0375] Example 18: Preparation of 3-chloro-N-[(1R,3S)-3-((6-chloro-2-(trifluoromethyl)quinolin-4-yl]amino)cyclohexyl]-1-[1-(methyldioxyylidene-λ6-thio)propyl]pyrazole-4-carboxamide (Compound 56)

[0376] The synthetic route of compound 56 is as follows:

[0377] Step 1: Synthesis of ethyl 3-chloro-1-(1-(methylsulfonyl)propyl)-1H-pyrazole-4-carboxylate

[0378] At room temperature, ethyl 3-chloro-1-((methylsulfonyl)methyl)-1H-pyrazole-4-carboxylate (44-3) (40.0 mg, 0.150 mmol) was dissolved in N,N-dimethylformamide (0.5 mL) and tetrahydrofuran (0.5 mL). Sodium hydroxide (6.0 mg, 0.150 mmol, 60%) was added under ice bath. After 20 minutes, iodoethane (23.4 mg, 0.150 mmol) was slowly added. The reaction solution was slowly heated to 25°C and stirred for 12 hours. After the reaction was completed, the mixture was quenched with saturated ammonium chloride solution (10 mL) under ice bath, and then extracted with ethyl acetate (20 mL). The organic phase was washed with water (10 mL) and saturated brine (10 mL) in sequence, and the organic phase was concentrated under reduced pressure to obtain compound 3-chloro-1-(1-(methylsulfonyl)propyl)-1H-pyrazole-4-carboxylic acid ethyl ester (56-1) (44.0 mg, yield 100%).

[0379] LC / MS (ESI) (m / z): 295.0 (M+H) + .

[0380] Step 2: 3-chloro-1-(1-(methylsulfonyl)propyl)-1H-pyrazole-4-carboxylic acid

[0381] Ethyl 3-chloro-1-(1-(methylsulfonyl)propyl)-1H-pyrazole-4-carboxylate (56-1) (44.0 mg, 0.150 mmol) was dissolved in tetrahydrofuran (0.4 mL) / methanol (0.2 mL) / water (0.2 mL) at room temperature. Lithium hydroxide monohydrate (31.0 mg, 0.750 mmol) was then added at room temperature. The reaction mixture was stirred at 25°C for 12 hours. After completion of the reaction, the pH was adjusted to 3 with dilute hydrochloric acid (1 mol / L) under ice-cooling. The mixture was then extracted with ethyl acetate (20 mL), washed sequentially with water (5 mL) and saturated brine (5 mL), and the organic phase was concentrated under reduced pressure to afford compound 3-chloro-1-(1-(methylsulfonyl)propyl)-1H-pyrazole-4-carboxylic acid (56-2) (40.0 mg, 100% yield).

[0382] LC / MS (ESI) (m / z): 267.0 (M+H) +

[0383] Step 3: 3-chloro-N-((1R,3S)-3-((6-chloro-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)-1-(1-(methylsulfonyl)propyl)-1H-pyrazole-4-carboxamide

[0384] 3-Chloro-1-(1-(methylsulfonyl)propyl)-1H-pyrazole-4-carboxylic acid (56-2) (40.0 mg, 0.150 mmol) was dissolved in N,N-dimethylformamide (1.0 mL) at room temperature, and then diisopropylethylamine (0.052 mL, 0.300 mmol), (7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (68.0 mg, 0.180 mmol) and (1R,3S)-3-{[6-chloro-2-(trifluoromethyl)quinolin-4-yl]amino}cyclohexan-1-amine (Intermediate A1) (52.0 mg, 0.150 mmol) were added in sequence. The reaction solution was stirred at 25°C for 12 hours. After the reaction was completed, the reaction solution was diluted with ethyl acetate (20 mL), washed with water (10 mL) and saturated brine (10 mL) in sequence, and the organic phase was concentrated under reduced pressure. The obtained crude product was purified by preparative high performance liquid chromatography (chromatographic column: YMC-Actus Triart C 18 ExRS-5μm 100*30mm; solvent: A=0.1% FA, B=acetonitrile; gradient: 30~80% 9min) to obtain compound 3-chloro-N-((1R,3S)-3-((6-chloro-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)-1-(1-(methylsulfonyl)propyl)-1H-pyrazole-4-carboxamide (56) (16.1 mg, yield 18.1%).

[0385] LC / MS (ESI) (m / z): 592.0 (M+H) + ;

[0386] 1 H NMR (400MHz, DMSO-d6): δ8.54(d,1H),8.39(d,1H),8.05(d,1H),7.86(d,1H),7.70(dd,1H),7.44(d,1H),6.89(s,1 H),5.69(dd,1H),3.96-3.79(m,2H),2.93(s,3H),2.22(m,3H),1.96-1.74(m,3H),1.53-1.22(m,4H),0.81(t,3H).

[0387] Example 19: Preparation of 3-chloro-N-((1R,3S)-3-((6-chloro-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)-1-(1-(ethylsulfonyl)ethyl)-1H-pyrazole-4-carboxamide (Compound 64)

[0388] The synthetic route of compound 64 is as follows:

[0389] Step 1: Synthesis of ethyl 3-chloro-1-hydroxymethyl-1H-pyrazole-4-carboxylate

[0390] To a solution of ethyl 3-chloro-1H-pyrazole-4-carboxylate (1.00 g, 5.73 mmol) in diethyl ether (5 mL) was added formaldehyde (0.16 mL, 5.73 mmol), and the reaction solution was stirred at 25° C. for 18 hours. After completion of the reaction, the reaction solution was quenched with saturated brine (100 mL), then extracted with ethyl acetate (100 mL×3). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to give a crude product which was separated and purified on a silica gel column (petroleum ether:ethyl acetate (V / V) = 100:0-50:50) to give ethyl 3-chloro-1-hydroxymethyl-1H-pyrazole-4-carboxylate (1.04 g, 88.7% yield).

[0391] LC-MS, M / Z(ESI):205.0[M+H] +

[0392] Step 2: Synthesis of ethyl 1-bromomethyl-3-chloro-1H-pyrazole-4-carboxylate

[0393] Phosphorus tribromide (2.13 g, 7.86 mmol) was added to a solution of ethyl 3-chloro-1-hydroxymethyl-1H-pyrazole-4-carboxylate (804 mg, 3.93 mmol) in diethyl ether (10 mL), and the reaction solution was stirred at 15° C. for 18 hours. After completion of the reaction, the reaction solution was quenched with water (100 mL), then extracted with ethyl acetate (100 mL×3). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to give a crude product which was separated and purified on a silica gel column (petroleum ether:ethyl acetate (V / V) = 100:0 to 50:50) to give ethyl 1-bromomethyl-3-chloro-1H-pyrazole-4-carboxylate (760 mg, 72.3% yield).

[0394] LC-MS, M / Z(ESI):267.0[M+H] +

[0395] Step 3: Synthesis of ethyl 3-chloro-1-((ethylthio)methyl)-1H-pyrazole-4-carboxylate

[0396] To a solution of ethyl 1-bromomethyl-3-chloro-1H-pyrazole-4-carboxylate (760 mg, 2.84 mmol) in acetonitrile (10 mL) was added sodium ethanethiolate (717 mg, 8.52 mmol), and the reaction mixture was stirred at 25° C. for 18 hours. After completion of the reaction, the reaction mixture was quenched with water (100 mL), then extracted with ethyl acetate (150 mL×3). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to give a crude product which was separated and purified on a silica gel column (petroleum ether:ethyl acetate (V / V) = 100:0 to 0:100) to give ethyl 3-chloro-1-((ethylthio)methyl)-1H-pyrazole-4-carboxylate (730 mg, 93.0% yield).

[0397] LC-MS, M / Z(ESI):249.0[M+H] +

[0398] Step 4: Synthesis of ethyl 3-chloro-1-((ethylsulfonyl)methyl)-1H-pyrazole-4-carboxylate

[0399] The compound 3-chloro-1-((ethylthio)methyl)-1H-pyrazole-4-carboxylic acid, ethyl ester (630 mg, 2.53 mmol) was dissolved in dichloromethane (10 mL), and m-chloroperbenzoic acid (1.31 g, 7.60 mmol) was added. The reaction solution was stirred at 25° C. for 2 h. After the reaction was completed, the reaction solution was poured into a saturated aqueous sodium thiosulfate solution (50 mL), and then extracted with ethyl acetate (100 mL×3). The organic phase was collected and washed with a saturated aqueous sodium carbonate solution (50 mL×3), then dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the compound 3-chloro-1-((ethylsulfonyl)methyl)-1H-pyrazole-4-carboxylic acid, ethyl ester (670 mg, 75.4% yield).

[0400] LC-MS, M / Z(ESI):281.0[M+H] +

[0401] Step 5: Synthesis of ethyl 3-chloro-1-(1-(ethylsulfonyl)ethyl)-1H-pyrazole-4-carboxylate

[0402] The compound 3-chloro-1-((ethylsulfonyl)methyl)-1H-pyrazole-4-carboxylic acid ethyl ester (670 mg, 2.39 mmol) was dissolved in anhydrous DMF (6 mL), cesium carbonate (1.56 g, 4.77 mmol) was added, and then iodomethane (678 mg, 4.77 mmol) was slowly added dropwise, and the reaction solution was stirred at 25 ° C for 18 h. After the reaction was completed, water (50 mL) was added to the reaction solution to quench the mixture, followed by extraction with ethyl acetate (100 mL × 3). The organic phase was collected and dried over anhydrous sodium sulfate, filtered and concentrated, and the residue was separated and purified by silica gel column chromatography (petroleum ether: ethyl acetate (V / V) = 100: 0 to 0: 100) to obtain compound 3-chloro-1-(1-(ethylsulfonyl)ethyl)-1H-pyrazole-4-carboxylic acid ethyl ester (630 mg, yield: 90.0%).

[0403] LC-MS, M / Z(ESI):295.0[M+H] +

[0404] Step 6: Synthesis of 3-chloro-1-(1-(ethylsulfonyl)ethyl)-1H-pyrazole-4-carboxylic acid

[0405] To a solution of ethyl 3-chloro-1-(1-(ethylsulfonyl)ethyl)-1H-pyrazole-4-carboxylate (330 mg, 1.12 mmol) in methanol (3 mL) / water (3 mL) / tetrahydrofuran (3 mL) was added lithium hydroxide (235 mg, 5.60 mmol), and the reaction solution was stirred at 10° C. for 1 hour. After completion of the reaction, the reaction solution was concentrated to give a crude product (260 mg), which was used directly in the next reaction.

[0406] LC-MS, M / Z(ESI):267.0[M+H] +

[0407] Step 7: Synthesis of 3-chloro-N-((1R,3S)-3-((6-chloro-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)-1-(1-(ethylsulfonyl)ethyl)-1H-pyrazole-4-carboxamide

[0408] To a solution of 3-chloro-1-(1-(ethylsulfonyl)ethyl)-1H-pyrazole-4-carboxylic acid (130 mg, 0.49 mmol) in dimethyl sulfoxide (3 mL) was added (1S,3R)-N 1-(6-chloro-2-(trifluoromethyl)quinolin-4-yl)cyclohexane-1,3-diamine (Intermediate A1) (168 mg, 0.49 mmol), 1-hydroxybenzotriazole (132 mg, 0.98 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (140 mg, 0.73 mmol) and N,N-diisopropylethylamine (189 mg, 1.46 mmol), the reaction solution was stirred at 10 ° C for 18 hours. After completion of the reaction, the reaction solution was diluted with saturated brine (50 mL), then extracted with ethyl acetate (50 mL×3), the organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to give a crude product which was purified by preparative high performance liquid chromatography (Xtimate C18, 21.2*250 mm, 5 μm; 10 mM NH4HCO3-ACN; 32-62; 60 mL / min) to give compound 3-chloro-N-((1R,3S)-3-((6-chloro-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)-1-(1-(ethylsulfonyl)ethyl)-1H-pyrazole-4-carboxamide (4.20 mg, yield 1.45%).

[0409] LC-MS, M / Z(ESI):592.2[M+H] +

[0410] 1 H NMR (400MHz, DMSO-d6): δ8.59(d,1H),8.49(d,1H),8.03(d,1H),7.90(d,1 H),7.74(dd,1H),7.49(d,1H),6.95(s,1H),5.98(q,1H),3.96-3.87(m,2H ),3.08(q,2H),2.54-2.52(m,1H),2.19-2.14(m,1H),1.99-1.88(m,2H),1 .83-1.80(m,3H),1.59-1.45(m,2H),1.41-1.24(m,2H),1.21-1.17(m,3H).

[0411] Example 20: Preparation of 5-chloro-N-((1R,3S)-3-((6-chloro-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)-1-((S-methylsulfonylimino)methyl)-1H-pyrazole-4-carboxamide (Compound 65)

[0412] The synthetic route of compound 65 is as follows:

[0413] Step 1: Synthesis of ethyl 5-chloro-1-((S-methylsulfonylimino)methyl)-1H-pyrazole-4-carboxylate

[0414] At room temperature, ethyl 5-chloro-1-((methylthio)methyl)-1H-pyrazole-4-carboxylate (500 mg, 2.13 mmol) was dissolved in anhydrous methanol (20 mL). Iodophenyldiacetic acid (2.06 g, 6.39 mmol) and ammonium carbonate (614 mg, 6.39 mmol) were then added. After the addition was complete, the reaction solution was stirred at room temperature for 4 hours. After TLC monitoring indicated complete reaction of the starting materials, stirring was stopped, the mixture was diluted with water (50 mL), and then extracted with ethyl acetate (30 mL x 3). The organic phase was collected and dried over anhydrous sodium sulfate. The organic phase was concentrated by distillation under reduced pressure, and the residue was purified by silica gel column chromatography (PE:EA = 100:10 (V / V)) to obtain ethyl 5-chloro-1-((S-methylsulfonylimino)methyl)-1H-pyrazole-4-carboxylate (500 mg, 88.34% yield).

[0415] LC-MS, M / Z(ESI):266.0[M+H] + .

[0416] Step 2: Synthesis of compound 5-chloro-1-((S-methylsulfonylimino)methyl)-1H-pyrazole-4-carboxylic acid (65-3)

[0417] At room temperature, ethyl 5-chloro-1-((S-methylsulfonylimino)methyl)-1H-pyrazole-4-carboxylate (270 mg, 1.02 mmol) was dissolved in a mixture of tetrahydrofuran / methanol / water (1:1:1) (3 mL). Hydrated lithium hydroxide (341 mg, 8.13 mmol) was then slowly added under ice-cooling conditions. The reaction mixture was stirred at room temperature for 2 hours. After TLC monitoring indicated complete reaction of the starting materials, the reaction mixture was diluted with water (10 mL), adjusted to pH 4 with dilute hydrochloric acid (1 M), and extracted with ethyl acetate (30 mL x 3). The organic phase was collected and dried over anhydrous sodium sulfate. The organic phase was concentrated by distillation under reduced pressure to obtain 5-chloro-1-((S-methylsulfonylimino)methyl)-1H-pyrazole-4-carboxylic acid (Compound 65-3) (220 mg, 91.10% yield).

[0418] LC-MS, M / Z(ESI):238.0[M+H] + .

[0419] Step 3: Synthesis of compound 5-chloro-N-((1R,3S)-3-((6-chloro-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)-1-((S-methylsulfonylimino)methyl)-1H-pyrazole-4-carboxamide

[0420] At room temperature, the compound 5-chloro-1-((S-methylsulfonylimino)methyl)-1H-pyrazole-4-carboxylic acid (70 mg, 0.29 mmol) was dissolved in anhydrous N,N-dimethylformamide (1 mL) and stirred in an ice bath for 10 minutes. Then, N,N-diisopropylethylamine (152 mg, 1.18 mmol) and N,N,N′,N′-tetramethyl-O-(7-azabenzotriazole-1-yl)uronium hexafluorophosphate (224 mg, 0.59 mmol) were added. Stirring was continued in an ice bath for 30 minutes. Then, (1S,3R)-N 1 -(6-chloro-2-(trifluoromethyl)quinolin-4-yl)cyclohexane-1,3-diamine (Intermediate A1) (111 mg, 0.32 mmol) was added and the mixture was slowly heated to room temperature and stirred for 6 hours. After TLC monitoring showed that the reaction of the starting materials was complete, stirring was stopped and the reaction solution was diluted with water (5 mL), then extracted with ethyl acetate (5 mL×3). The organic phase was collected and dried over anhydrous sodium sulfate. The organic phase was concentrated by distillation under reduced pressure, and the residue was separated and purified by silica gel column chromatography (DCM:MeOH=100:5 (V / V)) to obtain compound 5-chloro-N-((1R,3S)-3-((6-chloro-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)-1-((S-methylsulfonylimino)methyl)-1H-pyrazole-4-carboxamide (65) (50 mg, 30.20% yield).

[0421] LC-MS, M / Z(ESI):563.0[M+H] + .

[0422] 1 H NMR (400MHz, DMSO-d6): δ8.60(d,1H),8.16(d,J=0.7Hz,1H),8.07(d,1H),7.89(d,1H),7.74(dd,1H),7.48(d,1H), 6.94(s,1H),5.54(s,2H),4.16-4.12(m,1H),4.05-3.81(m,1H),2.97(s,3H),2.16(d,1H),2.00-1.79(m,3H),1.58 -1.27(m,5H).

[0423] Example 21: Preparation of 3-chloro-N-((1R,3S)-3-((6-fluoro-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)-1-(1-(methylsulfonyl)ethyl)-1H-pyrazole-4-carboxamide (Compound 77)

[0424] The synthetic route of compound 77 is as follows:

[0425] Step 1: Synthesis of tert-butyl ((1R,3S)-3-((6-fluoro-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)carbamate

[0426] At room temperature, the compound 4-chloro-6-fluoro-2-(trifluoromethyl)quinoline (300.00 mg, 1.20 mmol) and tert-butyl (1R,3S)-3-aminocyclohexyl)carbamate (386.40 mg, 1.80 mmol) were dissolved in anhydrous N,N-dimethylformamide (3 mL), and then N,N-diisopropylethylamine (621.37 mg, 4.80 mmol) was added, and the reaction solution was stirred at 100 ° C for 12 hours. After TLC monitoring showed that the reaction of the raw materials was completed, stirring was stopped, the reaction solution was cooled to room temperature, and then water (20 mL) was added to dilute it, and it was extracted with ethyl acetate (8 mL×3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered and concentrated, and the residue was separated and purified by silica gel column chromatography (petroleum ether:ethyl acetate (V / V)=100:10) to obtain the compound ((1R,3S)-3-((6-fluoro-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)carbamic acid tert-butyl ester (295 mg, yield 57.4%).

[0427] LC-MS, M / Z(ESI):428.6[M+H] +

[0428] Step 2: (1S, 3R)-N 1 Synthesis of 6-fluoro-2-(trifluoromethyl)quinolin-4-yl)cyclohexane-1,3-diamine hydrochloride

[0429] At room temperature, the compound ((1R,3S)-3-((6-fluoro-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)carbamic acid tert-butyl ester (250 mg, 0.58 mmol) was dissolved in dichloromethane (5 mL). The reaction solution was stirred in an ice bath for 10 minutes, and then a solution of hydrogen chloride in 1,4-dioxane (1 mol / L, 0.29 mL, 0.29 mmol) was slowly added dropwise. The reaction solution was slowly warmed to room temperature and stirred for 2 hours. After TLC monitoring showed that the reaction of the raw materials was complete, stirring was stopped and the reaction solution was concentrated under reduced pressure to remove the solvent to obtain compound (1S,3R)-N 1 -(6-Fluoro-2-(trifluoromethyl)quinolin-4-yl)cyclohexane-1,3-diamine hydrochloride (209 mg, crude product) was directly used in the next reaction.

[0430] LC-MS, M / Z(ESI):328.5[M+H] +

[0431] Step 3: Synthesis of 3-chloro-N-((1R,3S)-3-((6-fluoro-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)-1-(1-(methylsulfonyl)ethyl)-1H-pyrazole-4-carboxamide

[0432] At room temperature, compound 3-chloro-1-(1-(methylsulfonyl)ethyl)-1H-pyrazole-4-carboxylic acid (60 mg, 0.24 mmol) was dissolved in anhydrous N,N-dimethylformamide (2 mL), and then N,N-diisopropylethylamine (122.76 mg, 0.95 mmol) and N,N,N′,N′-tetramethyl-O-(7-azabenzotriazole-1-yl)uronium hexafluorophosphate (180.6 mg, 0.47 mmol) were added. The mixture was stirred at room temperature for 30 minutes, and then (1S,3R)-N 1 -(6-Fluoro-2-(trifluoromethyl)quinolin-4-yl)cyclohexane-1,3-diamine hydrochloride (93.3 mg, 0.28 mmol) was added, and the reaction solution was stirred at room temperature for 4 hours. After TLC monitoring showed that the reaction of the raw materials was complete, stirring was stopped, the reaction solution was cooled to room temperature, and then diluted with water (10 mL), extracted with ethyl acetate (5 mL×3), and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was separated and purified by silica gel column chromatography (petroleum ether:ethyl acetate (V / V) = 100:30) to obtain the compound 3-chloro-N-((1R,3S)-3-((6-fluoro-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)-1-(1-(methylsulfonyl)ethyl)-1H-pyrazole-4-carboxamide (48 mg, yield 35.97%).

[0433] LC-MS, M / Z(ESI):562.1[M+H] +

[0434] 1 H NMR(400MHz,DMSO-d6)δ8.47(d,1H),8.29(dd,1H),8.06(dd,1H),7.96(dd,1H),7.65(ddd,1H),7.28(d,1H),6.92(s,1H ),5.91(q,1H),4.02–3.81(m,2H),2.99(d,3H),2.17(d,1H),1.93(m,2H),1.82(dd,3H),1.61–1.24(m,3H),1.23(d,2H).

[0435] Step 4: 3-chloro-N-((1R,3S)-3-((6-fluoro-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)-1-((R)-1-(methylsulfonyl)ethyl)-1H-pyrazole-4-carboxamide and 3-chloro-N-((1R,3S)-3-((6-fluoro-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)-1-((S)-1-(methylsulfonyl)ethyl)-1H-pyrazole-4-carboxamide

[0436] Compound 3-chloro-N-((1R,3S)-3-((6-fluoro-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)-1-(1-(methylsulfonyl)ethyl)-1H-pyrazole-4-carboxamide (250 mg, 0.445 mmol) was purified by SFC (Daicel ChiralPak IC, 40 mm ID×250 mm, 10 μm, flowability: A = n-hexane, B = ethanol; slope: 30%, 1.68 h) to give compound 77-A (114.6 mg, yield 45.8%, retention time = 17.974 min) and compound 77-B (108.7 mg, yield 43.5%, retention time = 18.789 min).

[0437] Compound 77-A: LC-MS, M / Z (ESI): 562.2 [M+H] + ;

[0438] 1 H NMR (400MHz, DMSO-d6): δ8.47(s,1H),8.30(dd,1H),8.06(d,1H),7.97(dd,1H),7.71–7.61(m,1H),7.29(d,1H),6.93(s,1H ),5.92(q,1H),4.07–3.76(m,2H),2.99(s,3H),2.27–2.11(m,1H),2.06–1.88(m,2H),1.87–1.76(m,4H),1.57–1.28(m,4H).

[0439] Compound 77-B: LC-MS, M / Z (ESI): 562.2 [M+H] +

[0440] 1H NMR (400MHz, DMSO-d6): δ8.47(s,1H),8.30(dd,1H),8.06(d,1H),7.97(dd,1H),7.71–7.61(m,1H),7.29(d,1H),6.93(s,1H ),5.92(q,1H),4.07–3.76(m,2H),2.99(s,3H),2.27–2.11(m,1H),2.06–1.88(m,2H),1.87–1.76(m,4H),1.57–1.28(m,4H).

[0441] Example 22: Preparation of 3-chloro-N-((1R,3S)-3-((6-methyl-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)-1-(1-(methylsulfonyl)ethyl)-1H-pyrazole-4-carboxamide (Compound 80)

[0442] The synthetic route of compound 80 is as follows:

[0443] Step 1: Synthesis of tert-butyl ((1R,3S)-3-((6-methyl-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)carbamate

[0444] At room temperature, the compound 4-chloro-6-methyl-2-(trifluoromethyl)quinoline (500.00 mg, 2.04 mmol) and tert-butyl ((1R,3S)-3-aminocyclohexyl)carbamate (479.87 mg, 2.24 mmol) were dissolved in dimethyl sulfoxide (5 mL), and N,N-diisopropylethylamine (1.05 g, 8.14 mmol) was added. After the addition was completed, the reaction solution was stirred at 100 ° C for 8 hours. After TLC monitoring showed that the reaction of the raw materials was completed, stirring was stopped, the reaction solution was cooled to room temperature, and then water (30 mL) was added to dilute it, and extracted with ethyl acetate (10 mL×3). The organic phase was collected, dried over anhydrous sodium sulfate, filtered and concentrated, and the residue was separated and purified by silica gel column chromatography (petroleum ether:ethyl acetate (V / V)=80:20) to obtain compound ((1R,3S)-3-((6-methyl-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)carbamic acid tert-butyl ester (400 mg, yield 46.4%).

[0445] LC-MS, M / Z (ESI): 424.6 (M+H + )

[0446] Step 2: (1S, 3R)-N 1 Synthesis of 6-methyl-2-(trifluoromethyl)quinolin-4-yl)cyclohexane-1,3-diamine hydrochloride

[0447] At room temperature, the compound ((1R,3S)-3-((6-methyl-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)carbamic acid tert-butyl ester (400 mg, 0.94 mmol) was dissolved in dichloromethane (5 mL) and stirred at 0°C for 10 minutes. Then, a solution of hydrogen chloride in 1,4-dioxane (1 M, 0.47 mL, 0.47 mmol) was slowly added dropwise. After the addition was completed, the reaction solution was naturally warmed to room temperature and stirred for 2 hours. TLC monitoring showed that the reaction of the raw materials was complete, and stirring was stopped. The reaction solution was evaporated under reduced pressure to remove the solvent to obtain compound (1S,3R)-N 1 -(6-methyl-2-(trifluoromethyl)quinolin-4-yl)cyclohexane-1,3-diamine hydrochloride (350.00 mg, crude).

[0448] LC-MS, M / Z (ESI): 323.5 (M+H + )

[0449] Step 3: Synthesis of 3-chloro-N-((1R,3S)-3-((6-methyl-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)-1-(1-(methylsulfonyl)ethyl)-1H-pyrazole-4-carboxamide

[0450] At room temperature, compound 3-chloro-1-(1-(methylsulfonyl)ethyl)-1H-pyrazole-4-carboxylic acid (80 mg, 0.32 mmol) was dissolved in anhydrous N,N-dimethylformamide (2 mL), and then N,N-diisopropylethylamine (163.68 mg, 1.27 mmol) and N,N,N′,N′-tetramethyl-O-(7-azabenzotriazole-1-yl)uronium hexafluorophosphate (240.78 mg, 0.63 mmol) were added. The reaction solution was stirred at room temperature for 30 minutes, and then (1S,3R)-N 1 -(6-methyl-2-(trifluoromethyl)quinolin-4-yl)cyclohexane-1,3-diamine hydrochloride (170.89 mg, 0.47 mmol) was added and stirred at room temperature for 4 hours. After TLC monitoring showed that the reaction of the raw materials was complete, stirring was stopped, water (10 mL) was added to dilute, and extraction was performed with ethyl acetate (5 mL×3). The organic phase was collected and dried over anhydrous sodium sulfate, filtered and concentrated, and the residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate (V / V) = 100:40) to obtain the compound 3-chloro-N-((1R,3S)-3-((6-methyl-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)-1-(1-(methylsulfonyl)ethyl)-1H-pyrazole-4-carboxamide (39.6 mg, yield 22.4%).

[0451] LC-MS, M / Z (ESI): 558.1 (M+H + )

[0452] 1 H NMR(400MHz, CDCl3)δ8.23(s,1H),7.98(d,1H),7.53(d,1H),7.45(s,1H),6.73(s,1H),6.44(d,1H),5.31–5.24(m,1H),5.04(d,1H),4.23 –4.07(m,1H),3.82–3.64(m,1H),2.85(s,3H),2.61(d,1H),2.54(s,3H),2.32–2.23(m,1H),2.19(d,1H),1.98(d,4H),1.39–1.23(m,4H).

[0453] Example 23: Preparation of 3-chloro-N-((1R,3S)-3-((6-fluoro-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)-1-((methylsulfonyl)methyl)-1H-pyrazole-4-carboxamide (Compound 86)

[0454] The synthetic route of compound 86 is as follows:

[0455] At room temperature, compound 3-chloro-1-((methylsulfonyl)methyl)-1H-pyrazole-4-carboxylic acid (65.0 mg, 0.27 mmol) was dissolved in anhydrous N,N-dimethylformamide (2 mL), and then N,N-diisopropylethylamine (140.81 mg, 1.09 mmol) and N,N,N′,N′-tetramethyl-O-(7-azabenzotriazole-1-yl)uronium hexafluorophosphate (178.81 mg, 0.54 mmol) were added. The reaction solution was stirred at room temperature for 30 minutes, and then (1S,3R)-N 1-(6-Fluoro-2-(trifluoromethyl)quinolin-4-yl)cyclohexane-1,3-diamine hydrochloride (118.90 mg, 0.32 mmol) was added, and stirring was continued at room temperature for 4 hours. After TLC monitoring showed that the reaction of the raw materials was complete, stirring was stopped, and the reaction solution was diluted with water (10 mL). It was extracted with ethyl acetate (5 mL×3). The organic phase was collected and dried over anhydrous sodium sulfate. The organic phase was concentrated by distillation under reduced pressure. The residue was separated and purified by silica gel column chromatography (petroleum ether:ethyl acetate (V / V) = 100:30) to obtain the compound 3-chloro-N-((1R,3S)-3-((6-fluoro-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)-1-((methylsulfonyl)methyl)-1H-pyrazole-4-carboxamide (63.3 mg, 42.5% yield).

[0456] LC-MS, M / Z (ESI): 548.0 (M+H + )

[0457] 1 H NMR(400MHz,DMSO-d6)δ8.36(s,1H),8.29(dd,1H),8.13(d,1H),7.96(dd,1H),7.68–7.61(m,1H),7.29(d,1H),6.92(s, 1H),5.79(s,2H),4.06–3.79(m,2H),3.06(s,3H),2.16(d,1H),2.01–1.77(m,3H),1.60–1.42(m,2H),1.39–1.26(m,2H).

[0458] The preparation methods of the following compounds are described in Preparation Example 1.

[0459] Test Example 1: MRGPRX2 in vitro calcium flux assay

[0460] The determination of the antagonistic effect of the compounds on MRGPRX2 was carried out in a CHO stable cell line that highly expresses human MRGPRX2. 18 hours before the experiment, the cells were seeded at a certain density in a black-walled, transparent-bottomed plate containing DMEM / F12 (1:1) culture medium and incubated at 37°C, 5% CO2 for 18 hours. Then, the corresponding amount of dye solution was added to each well of the cells, and the cells were returned to the 37°C incubator for further incubation in the dark for 30 minutes, and then incubated at room temperature in the dark for 10 minutes. Then, different final concentrations of compounds were added to each well, and the cells were equilibrated for 20 minutes. Finally, a certain amount of C48 / 80 solution (sigma, lot: 0000197124) was added to the cells, and the fluorescence signal value was detected by FLIPR. The antagonistic effect (IC) of the compound was calculated using the software GraphPad Prism 8.0, with the compound concentration as the X-axis and the fluorescence signal value as the Y-axis. 50 The experimental results are shown in Table 1.

[0461] Table 1: MRGPRX2 calcium flux assay results

[0462] The results of the MRGPRX2 calcium flux test showed that the compound of the present invention has a good antagonistic effect on MRGPRX2.

[0463] Test Example 2: Compound toxicity test on hepatocytes

[0464] The toxicity test of the compounds on hepatocellular carcinoma was conducted on HepG2 (ATCC, HB-8065) ​​cells. Cell viability was measured using the CellTiter-Glo Luminescent Cell Viability Assay kit (Promega, G7573). The toxicity of the compounds was characterized by inhibition of HepG2 cell viability. HepG2 cells in the logarithmic phase were collected, the cell suspension concentration was adjusted, and 5000 cells / well were plated in a 96-well cell culture plate. The cells were incubated overnight in a cell culture incubator with 5% CO2 and 37°C. The next day, the medium was changed and different concentrations of compound solutions were added. A negative control group (cells + DMSO) and a blank control group (medium + DMSO) were set up at the same time. The cells were incubated in a cell culture incubator with 5% CO2 and 37°C for 72 hours. After the treatment, the kit instructions were followed and the luminescence signal values ​​in different wells were detected on the EnVision plate reader (2104). The inhibition of HepG2 cell viability by different concentrations of compounds was calculated according to the following formula, with the compound concentration as the X-axis and the inhibition rate as the Y-axis. The toxicity of the compound on HepG2 (IC 50 value).

[0465] The results of the hepatotoxicity test showed that the compound of the present invention exhibited good safety and low hepatotoxicity.

[0466] Test Example 3: Pharmacokinetics test in mice

[0467] Mouse pharmacokinetic studies were conducted using male ICR mice weighing 20-25 g and fasted overnight. Three mice were orally gavaged at 10 mg / kg. Blood was collected before dosing and 15, 30 minutes, and 1, 2, 4, 8, and 24 hours after dosing. Blood samples were centrifuged at 6800 g at 2-8°C for 6 minutes, and plasma was collected and stored at -80°C. Plasma was collected at each time point and mixed with 3-5 times the volume of acetonitrile solution containing the internal standard. The mixture was vortexed for 1 minute and centrifuged at 13,000 rpm at 4°C for 10 minutes. The supernatant was mixed with 3 times the volume of water, and an appropriate amount of the mixture was analyzed by LC-MS / MS. The main pharmacokinetic parameters were analyzed using a non-compartmental model using WinNonlin 7.0 software.

[0468] Table 2: Pharmacokinetic test results in mice

[0469] The results of the mouse pharmacokinetic test showed that the compound of the present invention exhibited excellent pharmacokinetic properties and good drugability.

[0470] Test Example 4: Pharmacokinetics test in rats

[0471] For the pharmacokinetic study in rats, male SD rats weighing 180-240 g were fasted overnight. Three rats were orally gavaged at 10 mg / kg. Blood was collected before dosing and 15, 30 minutes, and 1, 2, 4, 8, and 24 hours after dosing. Blood samples were centrifuged at 6800 g for 6 minutes at 2-8°C, and plasma was collected and stored at -80°C. Plasma was collected at each time point and mixed with 3-5 times the volume of acetonitrile solution containing the internal standard. The mixture was vortexed for 1 minute and centrifuged at 13,000 rpm at 4°C for 10 minutes. The supernatant was mixed with 3 times the volume of water, and an appropriate amount of the mixture was analyzed by LC-MS / MS. The main pharmacokinetic parameters were analyzed using a non-compartmental model using WinNonlin 7.0 software.

[0472] The results of the rat pharmacokinetic test showed that the compound of the present invention exhibited excellent rat pharmacokinetic properties and good drugability.

[0473] Test Example 5: Dog Pharmacokinetics Study

[0474] Canine pharmacokinetic studies were conducted using male Beagle dogs weighing 8-10 kg, fasted overnight. Three Beagle dogs were orally gavaged and administered 5 mg / kg. Blood was collected before dosing and 15, 30 minutes, and 1, 2, 4, 8, and 24 hours after dosing. Blood samples were centrifuged at 6800 g for 6 minutes at 2-8°C, and plasma was collected and stored at -80°C. Plasma was collected at each time point and mixed with 3-5 times the volume of acetonitrile solution containing the internal standard. The mixture was vortexed for 1 minute and centrifuged at 13,000 rpm at 4°C for 10 minutes. The supernatant was mixed with 3 times the volume of water, and an appropriate amount of the mixture was analyzed by LC-MS / MS. Key pharmacokinetic parameters were analyzed using a non-compartmental model using WinNonlin 7.0 software.

[0475] Table 5 Results of dog pharmacokinetic tests

[0476] The results of the canine pharmacokinetic test showed that the compound of the present invention exhibited excellent canine pharmacokinetic properties and good drugability.

[0477] Test Example 6: Determination of the inhibitory effect of compounds on BSEP bile efflux transporter

[0478] The inhibitory effect of compounds on the BSEP (Bile Salt Export Pump) bile efflux transporter was tested using vesicles expressing the human BSEP bile efflux transporter (GenoMembrane). The vesicles were preincubated with various concentrations of compound for 5 minutes. Negative control (NC) and positive control (PC) controls were also established: the NC control consisted of vesicles preincubated with blank buffer for 5 minutes at 37°C, while the PC control consisted of vesicles preincubated with a positive inhibitor for 5 minutes at 37°C. Subsequently, the cells were incubated with the probe substrate at 37°C for 5 minutes in the presence of ATP or AMP. The assay was terminated with pre-chilled Buffer B1 (10× Buffer B1 (Stopping and Washing Buffer): 100 mM Hepes-Tris, 1000 mM KNO3, 500 mM Sucrose). The test sample was transferred to a 96-well filter plate, filtered with a vacuum pump, and then washed repeatedly with 0.2 mL of pre-cooled Buffer B1 5 times. The vesicles on the filter plate were dissolved with 50 μL of 80% methanol. After collection, the filtrate was collected by centrifugation at 2000 rpm for 2 minutes. Repeat once, combine the two filtrates together, mix well, and obtain approximately 100 μL of filtrate. Pre-cooled methanol containing the internal standard was added and centrifuged at 12,000 rpm for 5 minutes. The supernatant was used for LC-MS / MS quantitative detection of the content of the transported substrate. The IC of the compound inhibiting the bile efflux transporter activity was calculated using the Prism software, with the compound concentration as the X-axis and the relative activity (% of NC) as the Y-axis. 50values ​​and inhibition rates.

[0479] The transport rate (activity) and relative activity under different conditions were calculated according to the following formula:

[0480] Substrate active transport rate (pmol / min / mg) =

[0481] The results of the BSEP bile efflux transporter inhibition test showed that the compound of the present invention had no obvious inhibitory effect on the BSEP bile efflux transporter and no risk of cholestatic toxicity.

[0482] Test Example 7: Thermodynamic Solubility Test

[0483] Prepare phosphate buffered saline (PBS) at pH 7.4. Accurately weigh the compound and add it to the prepared PBS at pH 7.4 to a concentration of 4 mg / mL. Shake the solution at 1000 rpm for 1 hour, then incubate at room temperature overnight. Centrifuge the incubated solution at 12000 rpm for 10 minutes to remove undissolved particles, and transfer the supernatant to a fresh centrifuge tube. After appropriate dilution of the supernatant, add acetonitrile containing the internal standard and quantify using a standard curve prepared in the same matrix.

[0484] The results of the thermodynamic solubility test show that the compound of the present invention has good thermodynamic solubility and good drugability.

[0485] Test Example 8: Human liver microsome stability test

[0486] The stability test for human liver microsomes was performed by incubating the compound with human liver microsomes in vitro. The test compound was first prepared as a 10 mM stock solution in DMSO solvent, and then the compound was diluted to 0.5 mM in acetonitrile. Human liver microsomes (Corning) were diluted with PBS to form a microsome / buffer solution, and this solution was used to dilute 0.5 mM of the compound to form a working solution. The working solution contained 1.5 μM compound and 0.75 mg / ml human liver microsomes. A deep-well plate was prepared, and 30 μL of the working solution was added to each well. The reaction was then initiated by adding 15 μL of preheated 6 mM NADPH solution and incubated at 37°C. The reaction was terminated by adding 135 μL of acetonitrile to the corresponding wells at 0, 5, 15, 30, and 45 minutes of incubation. After terminating the reaction with acetonitrile at the final 45-minute time point, the deep-well plate was vortexed for 10 minutes (600 rpm / min) and then centrifuged for 15 minutes. After centrifugation, the supernatant was collected and purified water was added in a 1:1 ratio. LC-MS / MS was then performed to obtain the ratio of the peak area of ​​the compound to the peak area of ​​the internal standard at each time point. The peak area ratios of the compound at 5, 15, 30, and 45 minutes were compared with the peak area ratio at 0 minute. The remaining percentage of the compound at each time point was calculated. T was calculated using Graphpad 5 software. 1 / 2 .

[0487] The results of the human liver microsome stability test show that the compound of the present invention exhibits excellent human liver microsome stability and good drugability.

[0488] Test Example 9: Inhibition test of compounds on cytochrome P450

[0489] Compounds were tested for their inhibitory potential against the cytochrome P450 (CYP450) isoform CYP3A4 (two substrates, midazolam and testosterone). Test compounds were prepared in DMSO to a 10 mM stock solution. The CYP3A4 inhibitor ketoconazole was prepared in DMSO to 10 mM, 2.5 mM, and 2.5 mM stock solutions. Test compounds and ketoconazole were diluted in acetonitrile to a 400-fold final concentration (compound: 10 μM, ketoconazole: 2.5 μM).

[0490] Potassium phosphate buffer (0.1 M, pH 7.4) was used to prepare 4 times the final concentration of NADPH cofactor (66.7 mg NADPH was added to 10 mL potassium phosphate buffer) and substrate. The final concentration of CYP3A4 substrate midazolam was 320 μM, and the final concentration of CYP3A4 substrate testosterone was 20 μM.

[0491] Prepare a 0.2 mg / mL human liver microsomal solution in potassium phosphate buffer on ice. Prepare test compound and control inhibitor solutions at 2x the final concentration in the human liver microsomal solution on ice. Add 30 μL of test compound and control inhibitor solution to each test well, along with 15 μL of substrate, in duplicate. Incubate the 96-well assay plate and NADPH solution at 37°C for 5 minutes. Add 15 μL of preheated 8 mM NADPH solution to the assay plate to initiate the reaction. Preincubate the CYP3A4 assay plate at 37°C for 5 minutes. Terminate the reaction by adding 120 μL of acetonitrile. After quenching, shake the plate on a shaker (IKA, MTS2 / 4) for 10 minutes (600 rpm / min) and then centrifuge for 15 minutes. After centrifugation, the supernatant was collected and purified water was added in a 1:1 ratio. LC-MS / MS was then performed to obtain the ratio of the compound peak area to the internal standard peak area. The peak area ratio of the compound was compared with the peak area ratio of the control inhibitor to calculate the inhibition rate.

[0492] Table 6 Inhibition data of 10 μM compounds on cytochrome P450 (CYP450) isoform CYP3A4 (two substrates midazolam and testosterone)

[0493] The results of the inhibition test of the compound on cytochrome P450 show that the compound of the present invention has no obvious inhibitory effect on CYP3A4 (two substrates, midazolam and testosterone) and has good drugability.

[0494] Test Example 10: Evans blue vascular permeability test

[0495] The Evans blue vascular permeability test was performed using C57 mice, with the left hind paw of the mouse as the model group and the right hind paw as the negative control group. Different doses of drugs were administered orally, and 2 hours after administration, a 0.4% Evans blue (Sigma-Aldrich, Lot#SHBP1253) solution was injected into the tail vein. 5-10 minutes later, a C48 / 80 (Sigma-Aldrich, Lot#C2313) solution was injected subcutaneously into the left hind paw, and a normal saline solution was injected into the right hind paw. The mice were killed 15 minutes later, and the thickness of the left and right hind paws of the mice was measured with a vernier caliper. The mouse feet were cut off, dried, and weighed. The paws were crushed, and the Evans blue in the paws was dissolved with acetone: normal saline (7:3). The absorbance was measured at 620 nm, and the absorbance per unit volume was calculated. The mouse ear swelling rate and the amount of Evans blue exuded per unit mass were calculated.

[0496] The results of the Evans blue vascular permeability test showed that the compound of the present invention exhibited good efficacy, could significantly inhibit C48 / 80-induced vascular leakage, and had good drugability.

[0497] The above is an exemplary description of the implementation methods of the technical solutions disclosed herein. It should be understood that the scope of protection of the present disclosure is not limited to the above-mentioned implementation methods. Any modifications, equivalent substitutions, improvements, etc. made by those skilled in the art within the spirit and principles of the present disclosure shall be included in the scope of protection of the claims of this application.

Claims

1. A compound represented by Formula I-A, a tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug thereof: Wherein, X 1 , X 2 , X 3 , X 4 , X 5 and X 6 each independently represents a ring atom; X 1 、 X 2 、 X 3 、 X 5 and X 6 each independently is N, CH2, CH or C; X 4 is C; X 1 Between X 2 and X, 5 between X 6 and X, the connecting bond is a single bond or a double bond; X 5 and X 6 The group fragment formed by connection wherein A and X 5 , X 6 together with the ring atoms form a 6- to 10-membered aryl group, a 3- to 11-membered heteroalkyl group or a 5- to 10-membered heteroaryl group; the heteroatoms are independently selected from one or more of N, O and S; and the A is further substituted by R c ; the R c substitution is one or more substitutions, and when there are multiple substituents R c , the substituents are the same or different; Ring B is -(CH2) 0-2 -C 3-12 cycloalkyl, -(CH2) 0-2 -3- to 10-membered heterocycloalkyl or -(CH2) 0-2 -5- to 10-membered heteroaryl; ring C is a 6- to 10-membered aryl or a 5- to 10-membered heteroaryl; R a and R c each independently is H, halogen, hydroxy, amino, cyano, carbonyl, oxo, C 1-6 alkyl, C 3-8 cycloalkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 haloalkyl, C 3-8 halocycloalkyl, C 1-6 alkylhydroxy, C 1-6 alkylcarbonyl, C 1-6 alkoxy, C 1-6 haloalkoxy or 3- to 10-membered heterocycloalkyl; the C 1-6 alkyl, C 3-8 cycloalkyl, C 2-6 alkynyl, C 1-6 haloalkyl, C 3-8 halocycloalkyl, C 1-6 alkylhydroxy, C 1-6 alkylcarbonyl, C 1-6 alkoxy and C 1-6 haloalkoxy are optionally substituted by one or more R d ; the R d is a substituent selected from the following: halogen, hydroxy, amino, nitro, cyano, carbonyl, oxo, carboxyl, C 1-6 alkyl, C 2-6 alkynyl, C 1-6 haloalkyl, C 1-6 alkylhydroxy, C 1-6 alkylcarbonyl, C 1-6 alkoxy, C 1-6 haloalkoxy; when the substituent R d is plural, the R d are the same or different; R b Halogen 、 Hydroxyl, amino, cyano, carbonyl, oxo, C 1-6 Alkyl, C 1-6 Alkylamino, C 3-8 Cycloalkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Haloalkyl, C 3-8 Halogenated cycloalkyl, C 1-6 Alkyl hydroxyl, C 1-6 Alkylcarbonyl, C 1-6 Alkoxy, C 1-6 Haloalkoxy, 3- to 10-membered heterocycloalkyl, -(CHR j ) 0-3 -S(=O)(=NR j )-R k 、-(CHR j ) 1-3 -S(O)2-R k 、-(CHR j ) 1-3 -OC(O)-N(R k )2、-(CHR j ) 0-3 -(N=)S(=O)(R k )2 or -(CHR j ) 0-3 -P(=O)(R k )2; said C 1-6 Alkyl, C 1-6 Alkylamino, C 3-8 Cycloalkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Haloalkyl, C 3-8 Halogenated cycloalkyl, C 1-6 Alkyl hydroxyl, C 1-6 Alkylcarbonyl, C 1-6 Alkoxy, C 1-6 Haloalkoxy, 3- to 10-membered heterocycloalkyl, -(CHR j ) 0-3 -S(=O)(=NR j )-R k 、-(CHR j ) 1-3 -S(O)2-R k 、-(CHR j ) 1-3 -OC(O)-N(R k )2、-(CHR j ) 0-3 -(N=)S(=O)(R k ) 2 and -(CHR j ) 0-3 -P(=O)(R k )2 is optionally substituted by one or more R d ; the R d is a substituent selected from the following: halogen, hydroxyl, amino, nitro, cyano, carbonyl, oxo, carboxyl, C 1-6 alkyl, C 2-6 alkynyl, C 1-6 haloalkyl, C 1-6 alkylhydroxyl, C 1-6 alkylcarbonyl, C 1-6 alkoxy, C 1-6 haloalkoxy; when the substituent R d is plural, the R d are the same or different; n is 1, 2, 3 or 4, and at least one of R b is -(CHR j ) 0-3 -S(=O)(=NR j )-R k 、-(CHR j ) 1-3 -S(O)2-R k 、-(CHR j ) 1-3 -OC(O)-N(R k )2、-(CHR j ) 0-3 -(N=)S(=O)(R k )2 or -(CHR j ) 0-3 -P(=O)(R k )2; R 1 、 R 2 and R 3 are each independently H, halogen, cyano, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 haloalkyl, C 3-8 cycloalkyl, halo-C 3-8 cycloalkyl, -OR f 、 -C(O)OR f 、 -OC(O)R f 、 -N(R f )2、 -N(R f )C(O)R f 、 -N(R f )S(O)2R f or -S(O)2R f ; said R f is selected from H, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 alkylamino, -(CH2)rR g , 6- to 10-membered aryl, C 3-8 cycloalkyl, 5- to 10-membered heteroaryl or 5- to 10-membered heterocycloalkyl, or two R f groups together with the atoms to which they are attached form a 5- to 11-membered heterocycloalkyl; said R g is H, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 3-8 cycloalkyl, 6- to 10-membered aryl, 5- to 10-membered heteroaryl or 5- to 10-membered heterocycloalkyl; R j and R k each independently is H, cyano, amino, C 1-6 alkyl, C 1-6 alkylamino, -(CH2) 0-3 C 3-8 cycloalkyl, -COC 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkyl substituted by hydroxy, a 5- to 10-membered heterocycloalkyl alone, or when the said R k is two, it forms a 3- to 10-membered heterocycloalkyl with the N, S or P atom to which it is attached; the amino, C 1-6 alkylamino, -(CH2) 0-3 C 3-8 cycloalkyl, -COC 1-6 alkyl, C 1- haloalkyl, C 1-6 alkyl substituted by hydroxy, a 5- to 10-membered heterocycloalkyl alone, and the 3- to 10-membered heterocycloalkyl formed by R k with the N, S or P atom to which it is attached is optionally substituted by one or more R m ; the said R m is a substituent selected from the following: halogen, hydroxy, amino, nitro, cyano, carbonyl, oxo, carboxyl, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkylhydroxy, C 1-6 alkylcarbonyl, C 1-6 alkoxy, C 1-6 haloalkoxy, C 3-8 cycloalkyl; when there are a plurality of substituents R m , the said R m are the same or different; m and r are each 0, 1, 2 or 3.

2. The compound represented by Formula I-A according to claim 1, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug, characterized in that, It has the structure shown in Formula II-A, Wherein, A and X 5 , X 6 together with the ring atoms form a 6- to 8-membered aryl group, a 3- to 8-membered heteroalkyl group, a 5-membered heteroaryl ring or a 6-membered heteroaryl ring; the heteroatoms are independently selected from one or more of N, O, and S; A is further substituted by R c ; the R c substitution is one or more substitutions, and when there are multiple substituents R c , the substituents are the same or different; ring C is a 6- to 10-membered aryl or a 5- to 8-membered heteroaryl; R a and R c each independently represents H, halogen, hydroxy, amino, cyano, carbonyl, oxo, C 1-6 alkyl, C 3-8 cycloalkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 haloalkyl, C 3-8 halocycloalkyl, C 1-6 alkylhydroxy, C 1-6 alkylcarbonyl, C 1-6 alkoxy, C 1-6 haloalkoxy or 3- to 8-membered heterocycloalkyl; R 1 、R 2 、R 3 each independently represents H, halogen, cyano, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 haloalkyl, C 3-8 cycloalkyl, halo-C 3-8 cycloalkyl; R b is halogen, hydroxy, amino, cyano, carbonyl, oxo, C 1-6 alkyl, C 1-6 alkylamino, C 3-8 cycloalkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 haloalkyl, C 3-8 halocycloalkyl, C 1-6 alkylhydroxy, C 1-6 alkylcarbonyl, C 1-6 alkoxy, C 1-6 haloalkoxy or 3- to 10-membered heterocycloalkyl, -(CHR j ) 0-3 -S(=O)(=NR j )-R k 、-(CHR j ) 1-3 -S(O)2-R k 、-(CHR j ) 1-3 -OC(O)-N(R k )2、-(CHR j ) 0-3 -(N=)S(=O)(R k )2、-(CHR j ) 0-3 -P(=O)(R k )2; the C 1-6 alkyl, C 1-6 alkylamino, C 3-8 cycloalkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1- 6haloalkyl, C 3-8 halocycloalkyl, C 1-6 alkylhydroxy, C 1-6 alkylcarbonyl, C 1-6 alkoxy, C 1-6 haloalkoxy, 3- to 10-membered heterocycloalkyl, -(CHR j ) 0-3 -S(=O)(=NR j )-R k 、-(CHR j ) 1-3 -S(O)2-R k 、-(CHR j ) 1-3 -OC(O)-N(R k )2、-(CHR j ) 0-3 -(N=)S(=O)(R k )2 and -(CHR j ) 0-3 -P(=O)(R k )2 is optionally substituted by one or more R d ; said R d is selected from the following substituents: halogen, hydroxy, amino, nitro, cyano, carbonyl, oxo, carboxy, C 1-6 alkyl, C 2-6 alkynyl, C 1-6 haloalkyl, C 1-6 alkylhydroxy, C 1-6 alkylcarbonyl, C 1-6 alkoxy, C 1-6 haloalkoxy; when the substituent R d is plural, said R d are the same or different; n is 1, 2, 3 or 4, and at least one of R b is -(CHR j ) 0-3 -S(=O)(=NR j )-R k 、-(CHR j ) 1-3 -S(O)2-R k 、-(CHR j ) 1-3 -OC(O)-N(R k )2、-(CHR j ) 0-3 -(N=)S(=O)(R k )2 or -(CHR j ) 0-3 -P(=O)(R k )2; R j and R k each independently is H, cyano, amino, C 1-6 alkyl, C 1-6 alkylamino, -(CH2) 0-3 C 3-8 cycloalkyl, -COC 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkyl substituted by a hydroxyl group, a single 5- to 10-membered heterocycloalkyl, or when said R k is two, together with the N, S or P atom to which it is attached forms a 3- to 10-membered heterocycloalkyl; said amino, C 1-6 alkylamino, -(CH2) 0-3 C 3-8 cycloalkyl, -COC 1-6 alkyl, C 1- haloalkyl, C 1-6 alkyl substituted by a hydroxyl group, a single 5- to 10-membered heterocycloalkyl, and R k and the 3- to 10-membered heterocycloalkyl formed with the N, S or P atom to which it is attached is optionally substituted by one or more R m ; said R m is a substituent selected from the following: halogen, hydroxyl, amino, nitro, cyano, carbonyl, oxo, carboxyl, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkylhydroxyl, C 1-6 alkylcarbonyl, C 1-6 alkoxy, C 1-6 haloalkoxy, C 3-8 cycloalkyl; when there are multiple substituents R m , said R m are the same or different; X 1 、X 2 、X 3 、X 4 、X 5 、X 6 、 The definitions of m and n are as described in claim 1.

3. The compound represented by Formula I-A according to claim 1, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug, characterized in that, selected from and / or, For and / or, ring B is For example and / or, said R b is F, Cl, CN, -CH3, cyclopropyl, -CHF2, -CH2CF3, -NH-CH3, -CH2C(CH3)2(OH), -S(=O)(=NH)-CH3, -S(=O)(=NH)-CH2CH3, -S(=O)(=NH)-(CH2CH2OH), -CH2-S(=O)(=NH)-CH3, -CH2-S(=O)(=NCH3)-CH3, -CHCH3-S(=O)(=NCH3)-CH3, -C(CH3)2-S(=O)(=NCH3)-CH3, -S(=O)(=NCH3)-CH3, -S(=O)(=NCOCH3)-CH3, -S(=O)(=NCN)-CH3, -CH2-S(O)2-CH3, -C(CH3)2-S(O)2-CH3, -CH(CH2CH3)-S(O)2-CH3, -CH2-S(O)2-CH(CH3)2, -(CH2)2-S(O)2-CH3, -CH(CH3)-S(O)2-CH3, -CH(CH3)-S(O)2-CH2CH3, -CH2-S(O)2-CH3, -CH2-S(O)2-CH2CH3, -CH2CH(CH3)-S(O)2-CH3, -CH(CH3)CH2-S(O)2-CH3, -CH2-OC(O)-N(CH3)2, -(CH2)2-OC(O)-N(CH3)2, -CH2CH(CH3)-OC(O)-N(CH3)2, -CH(CH3)CH2-OC(O)-N(CH3)2, -(CH2)2-OC(O)-N(CH3)(CH2CH2CF3), -(CH2)2-OC(O)-N(CH3)(CH2CH2OH), -(N=)S(=O)(CH3)2, -(N=)S(=O)(CH2CH3)2, -CH2-(N=)S(=O)(CH3)2, -P(=O)(CH3)2, -P(=O)(CH2CH3)2, 4. The compound represented by Formula I-A according to claim 1, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug, characterized in that, The R j and R k are each independently H, cyano, amino, -CH3, -CH2CH3, -CH2CH2CH3, -CH2CF3, -CH2CH2CF3, -NH-CH3, -COCH3, -CH(CH3)2, -CH2CH(CH3), -CH(CH3)CH2, -CH(CH3)-, -CH2CH2OH, or the R k forms with the N, S or P atom to which it is attached and / or, R d is methyl.

5. The compound represented by Formula I-A according to claim 1, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug, characterized in that, The R a and R c are each independently H, F, Cl, methyl, amino, oxo, -CF3, -CHF2, -CN; and / or, said R 1 、R 2 、R 3 are each independently H, methyl, -CH2CF3, -CF3, 6. The compound represented by Formula I-A, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug according to any one of claims 1 to 2, characterized in that, ring C is a 6- to 8-membered aryl or a 5- to 8-membered heteroaryl; and / or, ring C is a 5- to 6-membered nitrogen-containing heteroaryl, and the number of nitrogen atoms is 1, 2 or 3; and / or, ring C is phenyl, pyrazolyl or pyridyl; and / or, ring C is and / or For 7. The compound represented by Formula I-A, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug according to any one of claims 1 to 6, characterized in that, The compound is selected from the following structures: wherein, R 1 , R 2 , R 3 , R b , R c , and n independently of one another have the definitions described in any one of claims 1 to 6; And / or, the compound is selected from the following structures: Among them, R 1 , R 2 , R 3 , R b , R c , and n independently of each other have the definitions described in any one of claims 1 to 6.

8. The compound represented by Formula I-A, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug according to claim 1, characterized in that, The compounds include: Optionally, the pharmaceutically acceptable salt is trifluoroacetate.

9. The compound represented by Formula I-A according to claim 1, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug, characterized in that, The compounds include: and / or, the compound is selected from any of the following compounds:

10. A pharmaceutical composition, characterized in that, The pharmaceutical composition comprises: a compound represented by formula I-A as described in any one of claims 1-9, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug; and a pharmaceutically acceptable carrier.

11. Use of a compound represented by formula I-A as described in any one of claims 1-9, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug, or use of the pharmaceutical composition as claimed in claim 10, said use comprising: antagonizing MRGPRX2; and / or, preventing and / or treating diseases related to MRGPRX2; and / or, preparing a drug, pharmaceutical composition or preparation for antagonizing MRGPRX2, and / or preventing and / or treating diseases related to MRGPRX2.

12. The use according to claim 11, wherein, The diseases related to MRGPRX2 include: mast cell-related diseases, skin diseases (such as atopic dermatitis, contact dermatitis, urticaria, chronic spontaneous urticaria, inducible urticaria, pruritus), autoimmune diseases (such as allergy, mastocytosis, rheumatoid arthritis, asthma, ulcerative colitis and interstitial cystitis), nervous system diseases (such as pain).

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