MAS-related G protein receptor X2 antagonist and application thereof

Abstract

The invention provides a compound as shown in a formula I-A, and a tautomer, a stereoisomer, a hydrate, a solvate, a pharmaceutically acceptable salt or a prodrug thereof. The compound has a good antagonistic effect on the MAS-related G protein receptor X2. # imgabs0 #

Description

[0001] This application claims the right of priority from the earlier applications filed by the applicant:

[0002] a patent application No. 202410071703.7, titled "MAS-related G protein-coupled receptor X2 antagonist and uses thereof", filed with the China National Intellectual Property Administration on January 17, 2024;

[0003] a patent application No. 202410166244.0, titled "MAS-related G protein-coupled receptor X2 antagonist and uses thereof", filed with the China National Intellectual Property Administration on February 5, 2024;

[0004] a patent application No. 202410843343.8, titled "MAS-related G protein-coupled receptor X2 antagonist and uses thereof", filed with the China National Intellectual Property Administration on June 26, 2024;

[0005] a patent application No. 202411283294.3, titled "MAS-related G protein-coupled receptor X2 antagonist and uses thereof", filed with the China National Intellectual Property Administration on September 12, 2024;

[0006] The entire text of the said earlier applications is incorporated herein by reference. Technical Field

[0007] The present invention belongs to the field of medicine. Specifically, the present invention relates to a MAS-related G protein-coupled receptor X2 antagonist and uses thereof. Background Art

[0008] Mast cells (MCs) are tissue-resident immune cells originating from the hematopoietic lineage. MCs reside in various connective tissues and vascularized organs, with the largest number and highest density at the interface between the internal and external environments. They respond to foreign organisms and antigens, playing a sentinel role. These sites include the dermis, skeletal muscle, oral mucosa, gastrointestinal mucosa and submucosa, conjunctiva, alveoli and airways, and auricles. MCs in the dermis are often very close to blood vessels, nerves, and lymphatic vessels. After activation through the human high-affinity immunoglobulin E (IgE) receptor (FcεRI) and Mas-related G protein-coupled receptor X2 (MRGPRX2), MCs degranulate, releasing various bioactive substances (such as histamine and proteases) or de novo synthesizing new prostaglandins, leukotrienes, and some cytokines, which play a central role in host defense, inflammation, and allergic reactions.

[0009] Mas-related G protein-coupled receptors (MRGPRs) are divided into nine subfamilies according to different receptor members, namely: MRGPRA, B, C, D, E, F, G, H, and MRGPRX unique to primates. 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 mainly expressed in skin MCs, and its expression rate is even higher than that of Fcε-RI. MRGPRX2 can sense various endogenous or exogenous agonists, including polycationic compounds and polypeptides, to trigger mast cell degranulation reactions.

[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 of 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 various MRGPRX2 agonists, and block the release of trypsin and inflammatory factors in human mast cells. Oral administration of EP262 can efficiently inhibit agonist-induced mast cell degranulation and increased vascular permeability in MRGPRX2-KI mice, showing significant potential efficacy in the treatment of mast cell-related diseases. These all indicate that developing new MRGPRX2 antagonists for the treatment of various mast cell-related diseases is a promising direction.

[0011] Currently, there is no drug on the market as an MRGPRX2 antagonist. Therefore, developing 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 object of the present invention is to provide a new compound as an antagonist of MAS-related G protein receptor X2.

[0013] In the first aspect of the present invention, there is provided a compound represented by formula I-A, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug:

[0014]

[0015] Wherein,

[0016] X 1 、X 2 、X 3 、X 4 、X5 and X 6 Each independently represents a ring atom;

[0017] X 1 , X 2 , X 3 , X 5 and X 6 are each independently N, CH2, CH or C;

[0018] X 4 is C;

[0019] X 1 With X 2 Between, X 5 With X 6 The bond between them is a single bond or a double bond;

[0020] X 5 and X 6 The linked group fragment In the 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;

[0021] Ring B is -(CH2) 0-2 -C 3-12 Cycloalkyl, -(CH2) 0-2 -3- to 10-membered heterocycloalkyl or -(CH2) 0-2 -5-10 membered heteroaryl;

[0022] Ring C is a 6- to 10-membered aryl group or a 5- to 10-membered heteroaryl group;

[0023] R a and R c are each 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 Haloalkyl, C 3-8 Halogenated cycloalkyl, C 1-6 Alkyl hydroxyl, C 1-6 Alkylcarbonyl, C 1-6 Alkoxy, C 1-6haloalkoxy 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 is 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, 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, the R d are the same or different;

[0024] 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, 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)(Rk ) 2; The 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, 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 substituted by one or more R d ; The 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 multiple, the R d are the same or different;

[0025] n is 1, 2, 3 or 4, and at least one of the 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;

[0026] 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 、

[0027] -N(R f )S(O)2R f or -S(O)2R f ; the R f is selected from H, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 alkylamino,

[0028] -(CH2)rR g 、6 - 10 membered aryl, C 3-8 cycloalkyl, 5 - 10 membered heteroaryl or 5 - 10 membered heterocycloalkyl, or two R f groups together with the atoms to which they are attached form a 5 - 11 membered heterocycloalkyl; the R g is H, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 3-8 cycloalkyl, 6 - 10 membered aryl, 5 - 10 membered heteroaryl or 5 - 10 membered heterocycloalkyl;

[0029] R j and R k are each independently H, cyano, amino, C 1-6 alkyl, C 1-6 alkylamino, -(CH2) 0-3 C3-8 cycloalkyl, -COC 1-6 alkyl, C 1-6 haloalkyl, C substituted with a hydroxyl group 1-6 alkyl, 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 1-6 haloalkyl, C substituted with a hydroxyl group 1-6 alkyl, a single 5- to 10-membered heterocycloalkyl, and R k the 3- to 10-membered heterocycloalkyl formed with the N, S or P atom to which it is attached is optionally substituted with 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 the substituent R m is plural, said R m are the same or different;

[0030] m and r are each independently 0, 1, 2 or 3.

[0031] In a second aspect of the present invention, there is provided a compound of formula I, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug:

[0032]

[0033] wherein,

[0034] X 1 , X 2 , X 3 , X 4 , X 5 and X 6 each independently represents a ring atom;

[0035] X 1 , X 2 , X 3 , X 5 and X 6 each independently is N, CH2, CH or C;

[0036] X 4 is C;

[0037] X 1 and X 2 between, X 5 and X 6 The bond connected therebetween is a single bond or a double bond;

[0038] X 5 and X 6 The group fragment formed by connection In, A and X 5 , X 6 together with the ring atoms form a 6- to 10-membered aryl group, a 3- to 11-membered heterocycloalkyl 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 the substituents R c are multiple, the substituents are the same or different;

[0039] 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;

[0040] Ring C is a 6- to 10-membered aryl group or a 5- to 10-membered heteroaryl group;

[0041] R a and R c are each independently H, 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 haloalkyl, C 3-8 halocycloalkyl, C 1-6 alkylhydroxyl, 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 alkylhydroxyl, C 1-6 alkylcarbonyl, C 1-6 alkoxy and C1-6 The haloalkoxy group is optionally substituted by one or more R d groups; said 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 there are multiple substituents R d , said R d are the same or different;

[0042] R b is 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, 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 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 1-6The haloalkoxy group is optionally substituted by one or more Rs d ; said R d is a substituent selected from the following: 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 there are multiple substituents R d , said Rs d are the same or different;

[0043] n is 1, 2, 3 or 4, and at least one of the Rs 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;

[0044] Rs 1 , Rs 2 and Rs 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 ,

[0045] -N(R f )S(O)2R f or -S(O)2Rf ; said R f is selected from H, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 alkylamino,

[0046] -(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;

[0047] R j and R k are each independently H, cyano, amino, C 1-6 alkyl, C 3-8 cycloalkyl, C 1-6 haloalkyl, a single 5- to 10-membered heterocycloalkyl, or when said R k is two, they together with the attached N, S or P atom form a 5- to 10-membered heterocycloalkyl; said amino, C 1-6 alkyl, C 3-8 cycloalkyl, C 1-6 haloalkyl, a single 5- to 10-membered heterocycloalkyl, and R k together with the attached N, S or P atom form a 5- to 10-membered heterocycloalkyl which is optionally substituted by one or more R m ; said R m is selected from the following substituents: 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 the substituent R m is plural, said R m are the same or different;

[0048] m, and r are each 0, 1, 2 or 3.

[0049] In the present invention, the definitions of certain substituents in the compounds represented by Formula I-A or Formula I are as described below, and the definitions of the substituents not mentioned are as described in any of the above schemes.

[0050] In a preferred embodiment of the present invention, the compounds represented by Formula I-A are selected from the following structures:

[0051] 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 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; 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 group or a 5- to 8-membered heteroaromatic group; R a and R c are each 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 haloalkyl, C 3-8 halocycloalkyl, C 1-6 alkylhydroxy, C 1-6 alkylcarbonyl, C 1-6 alkoxy, C 1-6 haloalkoxy or a 3- to 8-membered heterocycloalkyl group; R 1 , R 2 , 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; 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, C1-6 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 1-6 haloalkyl, 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 with one or more R d ; the R d is selected from the following substituents: 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 1-6alkylcarbonyl, 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 the 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 are each 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 1-6 alkyl substituted by hydroxy, a 5- to 10-membered heterocycloalkyl alone, or when the R k are two, together with the N, S or P atom to which they are attached form a 3- to 10-membered heterocycloalkyl; the amino, 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, and the 3- to 10-membered heterocycloalkyl formed by R k and the N, S or P atom to which it is attached are optionally substituted by one or more R m ; the R m is selected from the following substituents: 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, C1-6 haloalkoxy, C 3-8 cycloalkyl; when the substituent R m is plural, said R m are the same or different; X 1 、X 2 、X 3 、X 4 、X 5 、X 6 、m and n are defined as described in the first aspect of the present invention.

[0052] In a preferred embodiment of the present invention, 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,

[0053] In a preferred embodiment of the present invention, the R b is F, Cl, -CH3, -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, -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,

[0054] In a preferred embodiment of the present invention, the R j and Rk each independently is H, cyano, amino, -CH3, -CH2CH3, -CH2CH2CH3, -CH2CF3, -CH2CH2CF3, -NH-CH3, -COCH3, -CH(CH3)2, -CH2CH(CH3), -CH(CH3)CH2, -CH(CH3)-, -CH2CH2OH, or said R k formed with the connected N, S or P atom

[0055] In a preferred embodiment of the present invention, said R j and R k each independently is H, cyano, amino, -CH3, -CH2CH3, -CH2CH2CH3, -CH2CF3, -CH2CH2CF3, -NH-CH3, -COCH3, -CH(CH3)2, -CH2CH(CH3), -CH(CH3)CH2, -CH(CH3)-, -CH2CH2OH, or said R k formed with the connected N, S or P atom

[0056] In a preferred embodiment of the present invention, the compound shown in formula I is selected from the following structures:

[0057] wherein, A and X 5 , X 6 ring atoms together form a 6-8 membered aryl, 3-8 membered heterocycloalkyl, 5 membered heteroaryl or 6 membered heteroaryl; the heteroatoms are independently selected from one or more of N, O and S; said A is further substituted by R c ; said R c substitution is one or more substitutions, and when the substituents R c are multiple, said substituents are the same or different; ring C is a 6-10 membered aryl or 5-8 membered heteroaryl; R a and R c each independently is H, 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 haloalkyl, C 3-8 halocycloalkyl, C 1-6 alkylhydroxyl, C 1-6 alkylcarbonyl, C 1-6alkoxy, C 1-6 haloalkoxy, or 3- to 8-membered heterocycloalkyl; R 1 , R 2 , 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; R b is 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, -(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 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 with one or more R d ; the R d is selected from the following substituents: halogen, hydroxy, amino, nitro, cyano, carbonyl, oxo, carboxyl, C 1-6 alkyl, C2-6 Alkynyl, C 1-6 Halogenated alkyl, C 1-6 Alkyl hydroxyl, C 1-6 Alkyl carbonyl, C 1-6 Alkoxy, C 1-6 Halogenated alkoxy; when the substituent R d is plural, the 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 are each independently H, cyano, amino, C 1-6 alkyl, C 3-8 cycloalkyl, C 1-6 halogenated alkyl, a separate 5- to 8-membered heterocycloalkyl, or when the said R k is two, it forms a 5- to 8-membered heterocycloalkyl with the connected N, S or P atom; the amino, C 1-6 alkyl, C 3-8 cycloalkyl, C 1-6 halogenated alkyl, a separate 5- to 8-membered heterocycloalkyl, and the 5- to 8-membered heterocycloalkyl formed by R k with the connected N, S or P atom are optionally substituted by one or more R m ; the said R m is selected from the following substituents: halogen, hydroxyl, amino, nitro, cyano, carbonyl, oxo, carboxyl, C 1-6 alkyl, C 1-6 halogenated alkyl, C 1-6 alkyl hydroxyl, C 1-6 alkyl carbonyl, C 1-6 alkoxy, C 1-6 halogenated alkoxy, C 3-8 cycloalkyl; when the substituent R m is plural, the said R mSame or different; X 1 , X 2 , X 3 , X 4 , X 5 , X 6 , the definitions of m and n are as described in the second aspect of the present invention.

[0058] In a preferred embodiment of the present invention, selected from

[0059] In a preferred embodiment of the present invention, is

[0060] In a preferred embodiment of the present invention, ring B is For example

[0061] In a preferred embodiment of the present invention, said R b 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,

[0062] In a preferred embodiment of the present invention, R d is methyl.

[0063] In a preferred embodiment of the present invention, the R a and R c are each independently H, F, Cl, methyl, amino, oxo, -CF3, -CHF2, -CN 。

[0064] In a preferred embodiment of the present invention, the R a and R c are each independently H, F, Cl, methyl, amino, oxo.

[0065] In a preferred embodiment of the present invention, the R 1 and R 2 and R 3 are each independently H, methyl, -CH2CF3, -CF3,

[0066] In a preferred embodiment of the present invention, ring C is a 6- to 8-membered aryl or a 5- to 8-membered heteroaryl.

[0067] In a preferred embodiment of the present invention, ring C is a 5- to 6-membered nitrogen-containing heteroaryl, and the number of nitrogen atoms is 1, 2, or 3.

[0068] In a preferred embodiment of the present invention, ring C is phenyl, pyrazolyl, or pyridyl.

[0069] In a preferred embodiment of the present invention, ring C is

[0070] In a preferred embodiment of the present invention, is

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

[0072]

[0073] wherein R 1 , R 2 , R 3 , R b , R c , and n each independently have the definitions described above.

[0074] In a preferred embodiment of the present invention, the compound is selected from the following structures:

[0075]

[0076] wherein R 1 , R 2, R 3 , R b , R c , n are each independently as defined above.

[0077] In a preferred embodiment of the present invention, the compound represented by formula I-A is selected from any of the following compounds:

[0078]

[0079]

[0080]

[0081]

[0082]

[0083]

[0084] In a preferred embodiment of the present invention, the pharmaceutically acceptable salt of the compound represented by formula I-A is trifluoroacetate. In a preferred embodiment of the present invention, the compound represented by formula I-A is selected from any of the following compounds:

[0085]

[0086]

[0087]

[0088]

[0089]

[0090]

[0091] In a preferred embodiment of the present invention, the compound represented by formula I-A is selected from any of the following compounds:

[0092]

[0093] In the third aspect of the present invention, there is provided a pharmaceutical composition, which comprises: a compound as described in the first aspect to the second aspect of the present invention, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug; and a pharmaceutically acceptable carrier.

[0094] In the fourth aspect of the present invention, there is provided the use of a compound, a tautomer, a stereoisomer, a hydrate, a solvate, a pharmaceutically acceptable salt or a prodrug as described in the first to second aspects of the present invention, or the use of the pharmaceutical composition as described in the third aspect of the present invention, and the use includes: antagonizing MRGPRX2; and / or, preventing and / or treating MRGPRX2-related diseases; and / or, preparing a drug, a pharmaceutical composition or a preparation for antagonizing MRGPRX2 and / or preventing and / or treating MRGPRX2-related diseases.

[0095] Preferably, the MRGPRX2-related diseases include: mast cell-related diseases.

[0096] Preferably, the MRGPRX2-related diseases include: skin diseases, autoimmune diseases, and nervous system diseases.

[0097] Preferably, the skin diseases are selected from atopic dermatitis, contact dermatitis, urticaria, and pruritus.

[0098] Preferably, the urticaria is selected from chronic spontaneous urticaria and inducible urticaria.

[0099] Preferably, the autoimmune diseases are selected from allergy, mastocytosis, rheumatoid arthritis, asthma, ulcerative colitis, and interstitial cystitis.

[0100] Preferably, the nervous system diseases are selected from pain.

[0101] In the fifth aspect of the present invention, there is provided a method for antagonizing MRGPRX2 or preventing and / or treating MRGPRX2-related diseases, including the step of administering to a subject in need a compound, a tautomer, a stereoisomer, a hydrate, a solvate, a pharmaceutically acceptable salt or a prodrug as described in the first to second aspects of the present invention.

[0102] The additional aspects and advantages of the present invention will be partly given in the following description, partly will become obvious from the following description, or will be understood through the practice of the present invention.

[0103] Term Definitions and Explanations

[0104] Unless otherwise specified, for the present invention application, including the terms and definitions recited in the specification and claims of this application are as follows. Those skilled in the art can understand that according to the convention used in the art, in the structural formula of this application, used to depict chemical bonds, which are the points where the chemical bonds are connected to the core structure or the backbone structure by a part or a substituent.

[0105] Unless otherwise specified, the term "pharmaceutically acceptable" refers to those compounds, materials, compositions, and / or dosage forms that, within the scope of sound medical judgment, are suitable for use in contact with the tissues of humans and animals without excessive toxicity, irritation, allergic response, or other problems or complications, commensurate with a reasonable benefit / risk ratio.

[0106] Unless otherwise specified, the term "pharmaceutically acceptable salt" refers to salts of pharmaceutically acceptable non-toxic acids or bases, including salts of inorganic acids and bases, organic acids and bases.

[0107] Unless otherwise specified, the term "pharmaceutical composition" denotes a mixture of one or more of the 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 the pharmaceutical composition is to facilitate the administration of the compound to an organism.

[0108] Unless otherwise specified, the term "prodrug" refers to a compound that can be converted into a biologically active compound of the present invention under physiological conditions or by solvolysis. The prodrugs of the present invention are prepared by modifying a functional group in the compound, which modification can be removed by conventional operations or in vivo to obtain the parent compound. Prodrugs include compounds formed by linking a hydroxyl group or an amino group in the compound of the present invention to any group, and when the prodrug of the compound of the present invention is administered to a mammalian individual, the prodrug is cleaved to form a free hydroxyl group and a free amino group, respectively.

[0109] Unless otherwise specified, the term "stereoisomer" refers to isomers produced by different spatial arrangements of atoms in a molecule, including cis-trans isomers, enantiomers, diastereoisomers, and conformational isomers.

[0110] Depending on the choice of starting materials and methods, the compounds of the present invention may exist in the form of one of the possible isomers or mixtures thereof, for example as pure enantiomers, or as mixtures of isomers, such as racemic and diastereomeric mixtures, depending on the number of asymmetric carbon atoms. When describing optically active compounds, the prefixes D and L or R and S are used to denote the absolute configuration of the molecule with respect to the chiral center(s) in the molecule. The prefixes D and L or (+) and (–) are symbols used to specify the rotation of plane-polarized light caused by the compound, where (–) or L indicates that the compound is levorotatory. Compounds with the prefix (+) or D are dextrorotatory. For 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 said isomers are commonly referred to as mixtures of enantiomers. A 50:50 mixture of enantiomers is called a racemic mixture or racemate, and such racemic mixtures or racemates may occur when there is no stereoselectivity or stereospecificity in a chemical reaction or process. Many geometric isomers of alkenes, C=N double bonds, etc. may also be present in the compounds described herein, and all such stable isomers are contemplated in the present invention. When the compounds described herein contain an alkene double bond, unless otherwise stated, such double bonds include the E and Z geometric isomers. If the compound contains a disubstituted cycloalkyl group, the substituents on the cycloalkyl group may be in the cis- or trans- configuration.

[0111] When depicting the bonds to a chiral carbon in the formulas of the present invention as straight lines, it should be understood that both the (R) and (S) configurations of the chiral carbon and, thus, both the enantiomerically pure compounds and mixtures thereof resulting therefrom are included within the scope of the general formula. The illustrations of racemates or enantiomerically pure compounds herein are from Maehr, J. Chem. Ed. 1985, 62:114-120. Unless otherwise stated, the absolute configuration of a stereocenter is represented by a wedge bond and a dashed bond.

[0112] The optically active (R)- or (S)-isomers can be prepared using chiral synthons or chiral preparations, or resolved using conventional techniques. The compounds of the present invention containing an asymmetrically substituted carbon atom can be isolated in optically active form or in racemic form. The resolution of the racemic mixtures of the compounds can be carried out by any of a number of methods known in the art. Exemplary methods include fractional crystallization using a chiral resolving acid, which is an optically active salt-forming organic acid. Suitable resolving agents for the fractional crystallization method are, for example, optically active acids such as tartaric acid, diacetyltartaric acid, dibenzoyltartaric acid, mandelic acid, malic acid, lactic acid or the D and L forms of various optically active camphorsulfonic acids such as β-camphorsulfonic acid. Other resolving agents suitable for the fractional crystallization method include α-methyl-benzylamine in stereoisomerically pure form (e.g., the S and R forms or diastereomerically pure form), 2-phenylglycol, norephedrine, ephedrine, N-methylephedrine, cyclohexylethylamine, 1,2-diaminocyclohexane, etc. The resolution of the racemic mixture can also be carried out by elution on a chromatographic column packed with an optically active resolving agent (e.g., dinitrobenzoyl-phenylglycine). High performance liquid chromatography (HPLC) or supercritical fluid chromatography (SFC) can be employed. The choice of the specific method, as well as the elution conditions and the selection of the chromatographic column, can be made by those skilled in the art according to the structure of the compound and the test results. Further, any enantiomer or diastereomer of the compounds described in the present invention can be obtained by stereoselective organic synthesis using optically pure starting materials or reagents of known configuration.

[0113] Unless otherwise specified, the term "tautomer" refers to functional group isomers resulting from the rapid migration of an atom within a molecule between two positions. The compounds of the present invention may exhibit tautomerism. Tautomeric compounds can exist in two or more interconvertible forms. Prototropic tautomers result from the migration of a hydrogen atom covalently bonded between two atoms. Tautomers generally exist in an equilibrium form, and attempting to isolate a single tautomer usually results in a mixture whose physicochemical properties are consistent with those of a mixture of the compounds. The position of the equilibrium depends on the chemical characteristics 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.

[0114] Unless otherwise indicated, a solid wedge bond and a dashed wedge bond are used to represent the absolute configuration of a stereocenter, and a solid straight bond and a dashed straight bond are used to represent the relative configuration of a stereocenter.

[0115] Unless otherwise specified, the term "solvate" refers to a stoichiometric or non-stoichiometric solvent that is bound to a compound or a salt thereof of the present invention by intermolecular non-covalent forces, and when the solvent is water, it is a hydrate.

[0116] For a drug or a pharmacological active agent, the term "effective amount" or "therapeutically effective amount" refers to a sufficient amount of the drug or agent that is non-toxic but can achieve the desired effect. For the oral dosage forms in the present invention, the "effective amount" of an active substance in the composition refers to the amount required to achieve the desired effect when used in combination with another active substance in the composition. The determination of the effective amount varies from person to person, depending on the age and general condition of the recipient, and also depends on the specific active substance. The appropriate effective amount in a particular case can be determined by those skilled in the art through routine tests.

[0117] Unless otherwise specified, the terms "active ingredient", "therapeutic agent", "active substance" or "active agent" refer to a chemical entity that can effectively treat a target disorder, disease or condition.

[0118] 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 variants of hydrogen, provided that the valence state of the particular atom is normal and the resulting compound is stable. When the substituent is a keto group (i.e., =O), it means that two hydrogen atoms are replaced. Keto substitution does not occur on an aromatic group.

[0119] In this application, "optional" or "optionally" means that the subsequent described event or condition may or may not occur, and the description includes both the occurrence and non-occurrence of the event or condition. For example, "optionally substituted aryl" means that the aryl is either substituted or unsubstituted, and the description includes both the substituted aryl and the unsubstituted aryl.

[0120] Unless otherwise specified, the term "C 1-6 alkyl" is used to denote a straight-chain or branched-chain saturated hydrocarbon group consisting of 1 to 6 carbon atoms. The C 1-6 alkyl includes C 1-5 、C 1-4 、C 1-3 、C 1-2 、C 2-6 、C 2-4 、C6 and C5 alkyl, etc.; it can be monovalent (such as methyl), divalent (such as methylene) or polyvalent (such as methine). Examples of C 1-6 alkyl 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, etc.

[0121] Unless otherwise specified, the term "C 1-3 alkyl" is used to denote straight-chain or branched saturated hydrocarbon groups consisting of 1 to 3 carbon atoms. The C 1-3 alkyl includes 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). Examples of C 1-3 alkyl include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), etc.

[0122] When used alone or as part of other substituents, the term "halo" can be used interchangeably with the term "halogen substituted".

[0123] Unless otherwise specified, "haloalkyl" or "halogen-substituted alkyl" refers to straight-chain and branched saturated aliphatic hydrocarbon groups containing a specific number of carbon atoms and substituted with one or more halogens, and can be "C 1-6 haloalkyl".

[0124] Unless otherwise specified, "C 2-6 alkenyl" is used to denote straight-chain or branched hydrocarbon groups consisting of 2 to 6 carbon atoms containing at least one carbon-carbon double bond, and the carbon-carbon double bond can be located at any position in the group. The C 2-6 alkenyl includes C 2-4 、C 2-3 、C4, C3, and C2 alkenyl, etc.; it can be monovalent, divalent, or polyvalent. Examples of C 2-6 alkenyl include, but are not limited to, vinyl, propenyl, butenyl, pentenyl, hexenyl, butadienyl, pentadienyl, hexadienyl, etc.

[0125] Unless otherwise specified, "C 2-6 alkynyl" is used to denote straight-chain or branched hydrocarbon groups consisting of 2 to 6 carbon atoms containing at least one carbon-carbon triple bond, and the carbon-carbon triple bond can be located at any position in the group. The C 2-6 alkynyl includes C 2-4 、C 2-3 、C4, C3, and C2 alkynyl, etc. It can be monovalent, divalent, or polyvalent. Examples of C 2-6 alkynyl include, but are not limited to, ethynyl, propynyl, butynyl, pentynyl, etc.

[0126] Unless otherwise specified, the term "C 1-6 alkoxy" denotes those alkyl groups containing 1 to 6 carbon atoms that are attached to the remainder of the molecule through an oxygen atom. The C 1-6 alkoxy includes C 1-4 、C 1-3 、C 1-2 、C2-6 , C 2-4 , C6, C5, C4, and C3 alkoxy groups, etc. C 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), pentyloxy (including n-pentyloxy, isopentyloxy, and neopentyloxy), hexyloxy, etc.

[0127] Unless otherwise specified, the term "C 1-3 alkoxy" refers to those alkyl groups containing 1 to 3 carbon atoms that are connected to the rest of the molecule through an oxygen atom. The C 1-3 alkoxy groups include C 1-2 , C 2-3 , C3, and C2 alkoxy groups, etc. C 1-3 Examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy (including n-propoxy and isopropoxy), etc.

[0128] Unless otherwise specified, the term "C 1-6 alkylamino" refers to those alkyl groups containing 1 to 6 carbon atoms that are connected to the rest of the molecule through an amino group. The C 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 groups, etc. C 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, etc.

[0129] Unless otherwise specified, the term "C 3-12 cycloalkyl" refers to a saturated cyclic hydrocarbon group composed of 3 to 12 carbon atoms, which includes monocyclic and bicyclic systems, where the bicyclic system includes spiro, fused, and bridged rings. The C 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 groups, etc.; it can be monovalent, divalent, or polyvalent. C 3-12Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, norbornyl, [2.2.2] bicyclooctane, etc.

[0130] Unless otherwise specified, the term "C 3-6 cycloalkyl" means a saturated cyclic hydrocarbon group composed of 3 to 6 carbon atoms, which is a monocyclic and bicyclic system. The C 3-6 cycloalkyl includes C 3-5 、C 4-5 and C 5-6 cycloalkyl, etc.; it can be monovalent, divalent or polyvalent. Examples of C 3-6 cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, etc.

[0131] Unless otherwise specified, C n-n+m or C n -C n+m includes any specific case of n to n + m carbons, such as C 1-12 includes C1, C2, C3, C4, C5, C6, C7, C8, C9, C 10 、C 11 、and C 12 ; it also includes any range within n to n + m, such as C 1-12 includes 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 etc.; Similarly, n - to n + m - membered means that the number of atoms in the ring is from n to n + m. For example, a 3 - 12 - membered ring includes a 3 - membered ring, 4 - membered ring, 5 - membered ring, 6 - membered ring, 7 - membered ring, 8 - membered ring, 9 - membered ring, 10 - membered ring, 11 - membered ring, and 12 - membered ring, and also includes any range within n to n + m, such as a 3 - 12 - membered ring includes a 3 - 6 - membered ring, 3 - 9 - membered ring, 5 - 6 - membered ring, 5 - 7 - membered ring, 6 - 7 - membered ring, 6 - 8 - membered ring, and 6 - 10 - membered ring, etc.

[0132] Unless otherwise specified, the term "aryl" refers to a monocyclic or polycyclic carbocyclic ring having 6 to 20 carbon atoms, where at least one ring is an aromatic ring. When one of the rings is a non - aromatic ring, the group can be connected through the aromatic ring or through the non - aromatic ring. Examples of aryl include, but are not limited to: phenyl, naphthyl, tetrahydronaphthyl, 2,3 - dihydroindenyl, biphenyl, phenanthryl, anthryl, and acenaphthylenyl, and can be "6 - 10 - membered aryl, 6 - 8 - membered aryl". The term "6 - 10 - membered aryl" refers to a monocyclic or polycyclic carbocyclic ring having 6 to 10 carbon atoms, where at least one ring is an aromatic ring.

[0133] Unless otherwise specified, the term "heterocycloalkyl" refers to a cycloalkyl in which one or more (in some embodiments, 1 to 3) carbon atoms are replaced by a heteroatom, such as but not limited to N, O, S, and P. The term "m-n membered heterocycloalkyl" or "Cm-Cn heterocycloalkyl" should be understood to represent a saturated, unsaturated, or partially saturated ring having from m to n atoms, wherein the heteroatoms are selected from N, O, S, P, preferably from N, O, or S, and can 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 separately represents a saturated cyclic group composed of 3 to 11 ring atoms. The term "6-8 membered heterocycloalkyl" alone or in combination with other terms separately represents a saturated cyclic group composed 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 atoms are optionally quaternized and the nitrogen and sulfur heteroatoms can be optionally oxidized (i.e., NO and S(O)p, where p is 1 or 2). It includes monocyclic and bicyclic systems, wherein the bicyclic systems include spirocycles, fused rings, and bridged rings. In addition, with respect to this "6-8 membered heterocycloalkyl", the heteroatom can occupy the position where the heterocycloalkyl is connected to the rest of the molecule. For example, 6-8 membered heterocycloalkyl includes but is not limited to 6-membered, 7-membered, 8-membered. Examples of 6-8 membered heterocycloalkyl include but are not limited to azetidinyl, oxetanyl, thietanyl, pyrrolidinyl, pyrazolidinyl, imidazolidinyl, tetrahydrothienyl (including tetrahydrothiophen-2-yl and tetrahydrothiophen-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.), dioxolanyl, dithiolanyl, isoxazolidinyl, isothiazolidinyl, 1,2-oxazinyl, 1,2-thiazinyl, hexahydropyridazinyl.

[0134] Unless otherwise specified, the term "heteroaryl ring" refers to a monocyclic or polycyclic carbocyclic 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, and 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-linked or N-linked, as long as it is possible). When one of the rings is a non-aromatic ring, the group may be linked 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, benzofuryl, quinolinyl, isoquinolinyl, oxazolyl, isoxazolyl, indolyl, pyrazinyl, pyridazinyl, pyridyl, pyrimidinyl, pyrrolyl, N-methylpyrrolyl, and tetrahydroquinoline. The term "heteroaryl ring" may be used interchangeably with the terms "heteroaromatic ring", "heteroaryl", or "heteroaryl group", and may be "5- to 10-membered heteroaryl, 5- to 8-membered heteroaryl".

[0135] Unless otherwise specified, the term "oxo" means that two hydrogens on the methylene group are replaced by oxygen, i.e., the methylene group is replaced by a carbonyl group, representing =O.

[0136] Unless otherwise specified, the term "halogen" or "halo group" means fluorine, chlorine, bromine, and iodine.

[0137] In addition, it should be noted that unless otherwise explicitly indicated, the description mode "…… independently" adopted in the present invention should be understood in a broad sense, which means that the described individuals are independent of each other and can independently be the same or different specific groups. More specifically, the description mode "…… independently" can either mean that among different groups, the specific options expressed by the same symbol do not affect each other, or it can also mean that within the same group, the specific options expressed by the same symbol do not affect each other.

[0138] In the present application, "pharmaceutically acceptable carrier" includes, but is not limited to, any adjuvant, carrier, excipient, glidant, sweetening agent, diluent, preservative, dye / colorant, flavoring agent, surfactant, wetting agent, dispersing agent, suspending agent, stabilizer, isotonic agent, solvent, or emulsifying agent that is permitted by the relevant government regulatory authorities for use in humans or livestock.

[0139] The term "treatment" refers to therapeutic treatment. When referring to a specific disease or disorder, treatment means: (1) alleviating one or more biological manifestations of the disease or disorder, (2) interfering with (a) one or more points in the biological cascade that causes or gives rise to the disorder or (b) one or more biological manifestations of the disorder, (3) improving one or more symptoms, effects, or side effects associated with the disorder, or one or more symptoms, effects, or side effects associated with the disorder or its treatment, or (4) slowing the progression of the disorder or one or more biological manifestations of the disorder.

[0140] The term "prevention" refers to a reduction in the risk of acquiring or developing a disease or disorder.

[0141] The term "patient" refers to any animal, preferably a mammal, that is about to receive or has received administration of the compound or composition according to an embodiment of the present invention. The term "mammal" includes any mammal. Examples of mammals include, but are not limited to, cows, horses, sheep, pigs, cats, dogs, mice, rats, rabbits, guinea pigs, monkeys, humans, etc., with humans being preferred.

[0142] The term "therapeutically effective amount" refers to the amount of a compound that, when administered to a patient, is sufficient to effectively treat a disease or disorder as described herein. The "therapeutically effective amount" will vary depending on the compound, the disorder and its severity, and the age of the patient to be treated, and can be adjusted by those skilled in the art as needed.

[0143] Based on common general knowledge in the art, the above preferred conditions can be combined arbitrarily to obtain various preferred embodiments of the present invention.

[0144] Beneficial effects

[0145] After extensive and in - depth research, the inventors of the present invention unexpectedly developed a compound or its pharmaceutically acceptable salt, and methods for its preparation and use. The present invention provides a compound represented by Formula I - A, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug. The compound of Formula I - A has a significant inhibitory effect on MRGPRX2, can be used as an antagonist of MAS - related G - protein - coupled receptor X2, and has high safety and drug - like properties. Detailed embodiments

[0146] The following further illustrates the present invention with specific embodiments. It should be understood that the following description is only the most preferred embodiment of the present invention and should not be considered as a limitation on the protection scope of the present invention. Based on a full understanding of the present invention, for the experimental methods without specific conditions noted in the following embodiments, they are generally carried out under conventional conditions or according to the conditions recommended by the manufacturer. Those skilled in the art can make non - essential modifications to the technical solutions of the present invention, and such modifications should be considered as being included within the protection scope of the present invention.

[0147] This application has the following definitions:

[0148] Symbol or unit:

[0149] IC 50 : Half maximal inhibitory concentration, referring to the concentration when the maximum inhibitory effect reaches half

[0150] M: mol / L. For example, n-butyllithium (14.56 mL, 29.1 mmol, 2.5 M n-hexane solution) represents an n-hexane solution of n-butyllithium with a molar concentration of 2.5 mol / L

[0151] N: Normality. For example, 2N hydrochloric acid represents a 2 mol / L hydrochloric acid solution

[0152] Reagent:

[0153] PPA: Polyphosphoric acid

[0154] DCE: 1,2 - Dichloroethane Intermediate A1: (1S,3R)-N 1 Synthesis of -(6-chloro-2-(trifluoromethyl)quinolin-4-yl)cyclohexane-1,3-diamine

[0155] The synthesis route of Intermediate A1 is as follows:

[0156]

[0157] The first step: Synthesis of 6-chloro-2-(trifluoromethyl)quinolin-4-ol (A1-3)

[0158]

[0159] At room temperature, p-chloroaniline (5 g, 39.37 mmol) was added to PPA (200 mmol, 75 mL, 5.0 eq), and then ethyl trifluoroacetoacetate (7.2 g, 39.37 mmol) was added. The reaction solution was heated to 140 °C and reacted for 12 hours. After the reaction was completed, the reaction solution was poured into ice water and extracted with ethyl acetate (4 * 50 mL). The obtained organic phase was dried with anhydrous sodium sulfate, filtered and concentrated to obtain the crude product of the compound 6-chloro-2-(trifluoromethyl)quinolin-4-ol (8.5 g, yield 87.6%), which was directly used for the next step of the reaction.

[0160] LC-MS, M / Z(ESI): 248.0[M+H] +

[0161] The second step: Synthesis of 4,6-dichloro-2-(trifluoromethyl)quinoline (A1-4)

[0162]

[0163] 6-Chloro-2-(trifluoromethyl)quinolin-4-ol (A1-3) (8.5 g, 34.5 mmol) was dissolved in DCE (100 mL). At 0 °C, phosphorus oxychloride (13.2 g, 86.25 mmol) was added. Subsequently, the reaction was heated to 80 °C and reacted for 12 hours. After the reaction was completed, the obtained crude product was dried by rotary evaporation and purified by silica gel column (PE:EA (V / V) = 100:2) to obtain 4,6-dichloro-2-(trifluoromethyl)quinoline (8.3 g, yield 91.6%).

[0164] LC-MS, M / Z (ESI): 265.9 [M+H] +

[0165] Step 3: Synthesis of tert-butyl ((1R,3S)-3-((6-chloro-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)carbamate (A1-6)

[0166]

[0167] 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), and then N,N-diisopropylethylamine (1.9 g, 15 mmol) was added. The reaction solution was heated to 130 °C and reacted for 12 hours. After the reaction was completed, the reaction solution was poured into ice water and extracted with ethyl acetate (4×50 mL). The obtained organic phase was dried over anhydrous sodium sulfate, filtered and dried by rotary evaporation. The obtained crude product was purified by silica gel column (PE:EA (V / V) = 2:1) to obtain tert-butyl ((1R,3S)-3-((6-chloro-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)carbamate (A1-6) (1.9 g, yield 96.3%).

[0168] LC-MS, M / Z (ESI): 444.1 [M+H] +

[0169] Step 4: Synthesis of (1S,3R)-N 1 -(6-chloro-2-(trifluoromethyl)quinolin-4-yl)cyclohexane-1,3-diamine (A1)

[0170]

[0171] At room temperature, tert-butyl ((1R,3S)-3-((6-chloro-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)carbamate (A1-6) (1 g, 2.3 mmol) was dissolved in 1,4-dioxane (20 mL), and then concentrated hydrochloric acid (0.5 mL) was added. The reaction mixture was stirred at room temperature for 1 hour. After the reaction was completed, the reaction mixture was adjusted to alkaline with saturated aqueous sodium bicarbonate, extracted with dichloromethane / methanol (4×50 mL), and the obtained organic phase was dried over anhydrous sodium sulfate, filtered and concentrated by rotary evaporation. The crude product was purified by silica gel column chromatography (DCM:MeOH (V / V) = 100:2) to obtain the compound (1S,3R)-N 1 -(6-chloro-2-(trifluoromethyl)quinolin-4-yl)cyclohexane-1,3-diamine (720 mg, yield 91.3%).

[0172] LC / MS (ESI) (m / z): 344 [M+H] +

[0173] 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)

[0174] The synthetic route of Compound 1 is shown as follows:

[0175]

[0176] First step: Synthesis of ethyl 1-(2-(methylsulfonyl)ethyl)-1H-pyrazole-4-carboxylate

[0177]

[0178] To a solution of ethyl 1H-pyrazole-4-carboxylate (500 mg, 3.57 mmol) in N,N-dimethylformamide (5 mL) were 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). Then the resulting mixture was stirred at 25 °C for 3 hours. The mixture was diluted with water (20 mL) and then extracted with ethyl acetate (15 mL×3). The combined organic layers were washed with saturated brine (30 mL), dried over sodium sulfate, and concentrated to obtain the crude product. The crude product was separated and purified by silica gel column chromatography (methanol:dichloromethane (V / V) = 100:1 - 100:5, gradient elution) to obtain ethyl 1-(2-(methylsulfonyl)ethyl)-1H-pyrazole-4-carboxylate (300 mg, yield 34.1%).

[0179] LC-MS, M / Z(ESI): 247.2 [M+H] +

[0180] Step 2: Synthesis of 1-(2-(methylsulfonyl)ethyl)-1H-pyrazole-4-carboxylic acid

[0181]

[0182] 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 h. The reaction solution was concentrated by rotary evaporation, and the crude product was separated and purified by 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, yield 86.5%).

[0183] LC-MS, M / Z(ESI): 219.2 [M+H] +

[0184] 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)

[0185]

[0186] To (1S,3R)-N 1To 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-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (199 mg, 0.52 mmol). 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 (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 obtain 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, yield 9.9%).

[0187] 1 H NMR (400 MHz, CD3OD): δ 8.54 (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).

[0188] LC-MS, M / Z (ESI): 544.2 [M + H] +

[0189] 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)

[0190] The synthetic route of Compound 5 is shown below:

[0191]

[0192] First step: Synthesis of ethyl 3-chloro-1-(2-(methylsulfonyl)ethyl)-1H-pyrazole-4-carboxylate

[0193]

[0194] At room temperature, ethyl 3-chloro-1H-pyrazole-4-carboxylate (0.500 g, 2.86 mmol) was dissolved in N,N-dimethylformamide (5 mL), then potassium carbonate (0.792 mg, 5.73 mmol), sodium iodide (4.00 mg, 0.286 mmol) and 2-(methylsulfonyl)ethyl bromide (589 mg, 3.15 mmol) were added. The reaction mixture was stirred at 25 °C for 3 h. After completion of the reaction, the reaction mixture was filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by column chromatography (PE:EA (V / V) = 10:1 to 1:1) to give ethyl 3-chloro-1-(2-(methylsulfonyl)ethyl)-1H-pyrazole-4-carboxylate (0.600 g, yield 74.8%).

[0195] LC / MS(ESI)(m / z):281.0(M+H) + .

[0196] Step 2: Synthesis of 3-chloro-1-(2-(methylsulfonyl)ethyl)-1H-pyrazole-4-carboxylic acid

[0197]

[0198] 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), then a solution of lithium hydroxide (128 mg, 5.34 mmol) in water (1 mL) was slowly added. After addition, the temperature was raised to 25 °C and the mixture was stirred for 12 h. After completion of the reaction, the reaction mixture was adjusted to pH 3 with 4N HCl solution in an ice bath, and a solid precipitated. The solid was filtered, and the filter cake was collected and dried to give 3-chloro-1-(2-(methylsulfonyl)ethyl)-1H-pyrazole-4-carboxylic acid (200 mg, yield 73.8%).

[0199] LC / MS(ESI)(m / z):253.0(M+H) +

[0200] 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

[0201]

[0202] 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), then diisopropylethylamine (56.0 mg, 0.440 mmol) and 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (86.0 mg, 0.230 mmol) were added, and the reaction mixture was stirred at room temperature for 2 h. After completion of the reaction, the reaction mixture was filtered, and the filtrate was concentrated in vacuo. The obtained crude product was purified by preparative liquid chromatography (column: YMC-Triart Prep C18 7 μm 30 mm × 40 cm; solvent: A = 0.1% aqueous ammonia, B = acetonitrile; gradient: 30% - 87%, 9 min) 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%).

[0203] LC / MS(ESI)(m / z): 578.0(M+H) + ;

[0204] 1 1H NMR(400 MHz, CDCl3) δ 8.07(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).

[0205] 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)

[0206] The synthetic route of Compound 7 is shown as follows:

[0207]

[0208] Step 1: Synthesis of 3-chloro-1-((methylsulfonyl)methyl)-1H-pyrazole-4-carboxylic acid (7-2)

[0209]

[0210] Dissolve ethyl 3-chloro-1-((methylsulfonyl)methyl)-1H-pyrazole-4-carboxylate (7-1) (80 mg, 0.30 mmol) in a mixed solution of methanol (2 mL) and water (0.8 mL), then add sodium hydroxide (96 mg, 2.40 mmol), and stir the reaction solution at room temperature for 2 h. After monitoring by TLC shows that the raw materials have reacted completely, stop stirring, distill off methanol under reduced pressure, add water (5 mL) to dilute the residue, then adjust the pH to 2 with dilute hydrochloric acid (2 mol / L), extract with ethyl acetate (15 mL × 3), collect the organic phase and dry it with anhydrous sodium sulfate, distill off the organic phase under reduced pressure to concentrate, and obtain 3-chloro-1-((methylsulfonyl)methyl)-1H-pyrazole-4-carboxylic acid (70 mg, yield 97.79%). The obtained crude product can be directly used for the next reaction.

[0211] 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)

[0212]

[0213] Dissolve 3-chloro-1-((methylsulfonyl)methyl)-1H-pyrazole-4-carboxylic acid (7-2) (70 mg, 0.29 mmol) in anhydrous DMF (2 mL), stir the reaction solution at 0 °C for 10 min, then add DIPEA (114 mg, 0.88 mmol) and HATU (223 mg, 0.59 mmol), after stirring at 0 °C for 30 min, add (1S,3R)-N 1-(6-Chloro-2-(trifluoromethyl))-quinolin-4-yl)cyclohexane-1,3-diamine (Intermediate A1) (111 mg, 0.32 mmol), and the resulting reaction solution was slowly warmed to room temperature and stirred for an additional 6 h. After monitoring by TLC showed that the raw materials had reacted completely, stirring was stopped, and the reaction was quenched by adding water (5 mL) to the reaction solution. Then, it was extracted with ethyl acetate (15 mL × 3), the organic phase was collected and dried over anhydrous sodium sulfate, and the organic phase was concentrated by distillation under reduced pressure. 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, yield 28.39%).

[0214] LC-MS, M / Z (ESI): 564.0 (M+1)

[0215] 1 H NMR (400 MHz, DMSO-d6) δ 8.59 (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). 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)

[0216] The synthetic route of Compound 8 is as follows:[[]]

[0217]

[0218] The first step: Synthesis of ethyl 1-(2-hydroxyethyl)-1H-pyrazole-4-carboxylate

[0219]

[0220] At 0 °C, a solution of 2-diazo-1-ol (5.28 g, 69.4 mmol) in ethanol (40 mL) was added dropwise to a solution of ethyl 2-formyl-3-oxopropionate (10.0 g, 69.4 mmol) and 4A molecular sieve (6.00 g, 69.4 mmol) in ethanol (40 mL). Then the resulting mixture was stirred at 25 °C for 13 h. The reaction solution was filtered through diatomaceous earth, and the filtrate was concentrated in vacuo. The crude product was separated and purified by reversed-phase column chromatography (water: acetonitrile (V / V) = 100:0 - 10:1, gradient elution) to obtain ethyl 1-(2-hydroxyethyl)-1H-pyrazole-4-carboxylate (6.50 g, yield 50.9%).

[0221] LC-MS, M / Z(ESI): 185.2 [M+H] +

[0222] Step 2: Ethyl 1-(2-((dimethylcarbamoyl)oxy)ethyl)-1H-pyrazole-4-carboxylate

[0223]

[0224] 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). Then the resulting mixture was stirred at 25 °C for 1 h under nitrogen protection, and subsequently dimethylamine (0.73 g, 16.4 mmol) was slowly added to the reaction solution through a syringe. The resulting mixture was stirred at 25 °C for 12 h. The reaction mixture was concentrated in vacuo, and then added to dichloromethane (20 mL) and hydrochloric acid (10 mL, 1N aqueous solution). The layers were 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 the crude product. The crude product was separated and purified by silica gel column chromatography (methanol: dichloromethane (V / V) = 100:1 - 100:5, gradient elution) to obtain ethyl 1-(2-((dimethylcarbamoyl)oxy)ethyl)-1H-pyrazole-4-carboxylate (900 mg, yield 43.3%).

[0225] LC-MS, M / Z(ESI): 256.2 [M+H] +

[0226] Step 3: 1-(2-((dimethylcarbamoyl)oxy)ethyl)-1H-pyrazole-4-carboxylic acid

[0227]

[0228] 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 mixture was stirred at 25 °C for 3 h. After concentration under reduced pressure, the residue was purified by reverse-phase column chromatography (water:acetonitrile (v / v) = 100:0 - 100:1) to give 1-(2-((dimethylcarbamoyl)oxy)ethyl)-1H-pyrazole-4-carboxylic acid (240 mg, yield 59.9%).

[0229] LC-MS, M / Z(ESI): 228.2[M+H] +

[0230] 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)

[0231]

[0232] To a solution of (1S,3R)-N 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) was 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-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (111 mg, 0.29 mmol). The resulting mixture was then stirred under nitrogen at 25 °C for 12 h. The reaction mixture was purified by preparative high performance liquid chromatography (column: Kinetex EVO C18, 21.2 * 250 mm, 5 μm; 0.05% NH3.H2O - ACN; 37 - 67%; 20 mL / 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, yield 27.6%).

[0233] LC-MS, M / Z(ESI): 553.2[M+H] +

[0234] 11H NMR (400 MHz, CD3OD): δ 8.38 (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).

[0235] Example 5: Preparation of N-(1R,3S)-3-((6-chloro-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)-4-(dimethylphosphoryl)benzamide (Compound 32)

[0236] The synthetic route of Compound 32 is as follows:

[0237]

[0238] The first step: Synthesis of methyl 4-(dimethylphosphoryl)benzoate

[0239]

[0240] Dissolve methyl 4-iodobenzoate (2.00 g, 7.63 mmol) in dimethyl sulfoxide (20 mL). Under nitrogen protection, add 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) in sequence. The reaction is stirred at 25 °C for 10 minutes, then add a solution of dimethylphosphine oxide (0.89 g, 11.4 mmol) in dimethyl sulfoxide (1 mL). The reaction solution is reacted at 130 °C for 1 hour under nitrogen protection. Cool the reaction solution to room temperature, dilute with water (100 mL), and then extract with ethyl acetate (100 mL × 3). Combine the organic phases, wash with saturated brine (50 mL), dry over anhydrous sodium sulfate, filter and concentrate to obtain the crude product. The crude product is purified by a reverse-phase chromatographic column (water:acetonitrile (V / V) = 100:1 to 2:1, gradient elution) to obtain methyl 4-(dimethylphosphoryl)benzoate (1.50 g, yield 87.9%).

[0241] LC-MS, M / Z (ESI): 213.2 [M + H] + .

[0242] The second step: Synthesis of 4-(dimethylphosphoryl)benzoic acid

[0243]

[0244] Methyl 4-(dimethylphosphoryl)benzoate (1.00 g, 4.48 mmol) was dissolved in water (2 mL) and methanol (20 mL), and then lithium hydroxide (3.76 g, 89.5 mmol) was added. The reaction mixture was reacted at 25 °C for 1 hour. The reaction solution was adjusted to pH = 2 with hydrochloric acid (2 M), and then washed with ethyl acetate (10 mL × 3). The aqueous phase was concentrated to obtain the crude product. The crude product was purified by reverse-phase chromatography column (water:acetonitrile (V / V) = 100:1 - 10:3, gradient elution) to obtain 4-(dimethylphosphoryl)benzoic acid (750 mg, yield 82.6%).

[0245] LC-MS, M / Z (ESI): 199.4 [M+H] +

[0246] Step 3: N-(1R,3S)-3-((6-chloro-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)-4-(dimethylphosphoryl)benzamide (Compound 32)

[0247]

[0248] 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) were added in sequence. The reaction mixture was stirred at 25 °C for 18 hours under nitrogen protection. 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 the crude product. It was separated and purified by silica gel column (methylene chloride: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, yield 10.8%).

[0249] 11H NMR (400 MHz, CD3OD): δ 8.38 (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 - 3.74 (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).

[0250] LC - MS, M / Z (ESI): 523.8 [M + H] +

[0251] Example 6: Preparation of N - ((1R,3S) - 3 - ((6 - chloro - 2 - (trifluoromethyl)quinolin - 4 - yl)amino)cyclohexyl)-4-(dimethylphosphoryl)benzamide (Compound 33)

[0252] The synthetic route of Compound 33 is as follows:

[0253]

[0254] First step: Synthesis of methyl 4 - bromo - 3 - ((tert - butoxycarbonyl)amino)benzoate

[0255]

[0256] To a solution of methyl 3 - amino - 4 - bromobenzoate (5.00 g, 21.7 mmol) in dichloromethane (50 mL) were successively 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). The reaction mixture was stirred at 25 °C for 12 hours. The reaction was quenched with saturated brine (50 mL), then extracted with dichloromethane (50 mL × 3). The organic layers were combined, dried over anhydrous sodium sulfate and concentrated to obtain the crude product, which was purified by silica gel column chromatography (petroleum ether:ethyl acetate (V / V) = 1:0 - 10:1, gradient elution) to give methyl 4 - bromo - 3 - ((tert - butoxycarbonyl)amino)benzoate (2.01 g, yield 27.9%).

[0257] Second step: Synthesis of methyl 3 - amino - 4 - (dimethylphosphoryl)benzoate

[0258]

[0259] To a solution of methyl 4-bromo-3-((tert-butoxycarbonyl)amino)benzoate (1.90 g, 5.75 mmol) in dimethyl sulfoxide (15 mL) were successively added palladium acetate (129 mg, 0.58 mmol), 1,3-bis(diphenylphosphino)propane (475 mg, 1.15 mmol), potassium phosphate (3.66 mg, 17.3 mmol). The reaction mixture was stirred at 25 °C for 5 minutes under nitrogen protection, and then a solution of dimethylphosphine oxide (665 mg, 8.63 mmol) in dimethyl sulfoxide (5 mL) was added. The reaction 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 column (water:acetonitrile (V / V)=100:0 - 5:3, gradient elution) to obtain methyl 3-amino-4-(dimethylphosphoryl)benzoate (800 mg, yield 61.2%).

[0260] LC-MS, M / Z(ESI): 228.0[M + H] +

[0261] Step 3: Synthesis of methyl 4-(dimethylphosphoryl)-3-(methylamino)benzoate

[0262]

[0263] To a solution of methyl 3-amino-4-(dimethylphosphoryl)benzoate (500 mg, 2.20 mmol) in methanol (5 mL) were successively added acetic acid (0.01 mL, 0.22 mmol), paraformaldehyde (330 mg, 11.0 mmol), sodium cyanoborohydride (415 mg, 6.60 mmol). The reaction was stirred at 50 °C for 12 hours. Water (5 mL) was added to quench the reaction, and the mixture was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by silica gel column (methylene chloride:methanol (V / V)=100:1 - 10:1, gradient elution) to obtain methyl 4-(dimethylphosphoryl)-3-(methylamino)benzoate (450 mg, yield 74.6%).

[0264] LC-MS, M / Z(ESI): 242.2[M + H] +

[0265] Step 4: 4-(Dimethylphosphoryl)-3-(methylamino)benzoic acid

[0266]

[0267] 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). The reaction mixture was stirred at 25 °C for 2 h. The reaction mixture was concentrated in vacuo, and the crude product was purified by reverse-phase chromatography (water:acetonitrile (V / V) = 100:1 - 10:1, gradient elution) to give 4-(dimethylphosphoryl)-3-(methylamino)benzoic acid (200 mg, yield 84.9%).

[0268] LC-MS, M / Z (ESI): 228.0 [M+H] +

[0269] Step 5: N-((1R,3S)-3-((6-Chloro-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)-4-(dimethylphosphoryl)benzamide

[0270]

[0271] To a solution of 4-(dimethylphosphoryl)-3-(methylamino)benzoic acid (150 mg, 0.66 mmol) in dimethyl sulfoxide (5 mL) were successively 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). The reaction mixture was stirred at 25 °C for 2 h. The reaction mixture 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, and concentrated to give the crude product. The crude product was purified by preparative high-performance liquid chromatography (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, yield 20.3%).

[0272] 11H NMR (400 MHz, DMSO-d6): δ 8.61 (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).

[0273] LC-MS, M / Z (ESI): 553.2 (M + H) +

[0274] Example 7: Preparation of N-((1R,3S)-3-((6-chloro-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)-3-(dimethylphosphoryl)benzamide (Compound 36)

[0275] The synthetic route of Compound 36 is as follows:

[0276]

[0277] First step: Synthesis of methyl 3-(dimethylphosphoryl)benzoate

[0278]

[0279] Dissolve methyl 3-iodobenzoate (3.00 g, 11.4 mmol) in dimethyl sulfoxide (30 mL). Under nitrogen protection, add 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) in sequence. Stir the reaction solution at 25 °C for 10 minutes under nitrogen protection, then add a solution of dimethylphosphine oxide (1.34 g, 17.2 mmol) in dimethyl sulfoxide (3 mL). Stir this mixture at 130 °C for 2 hours under N2 protection. Cool the reaction mixture to room temperature, dilute with water (100 mL), and then extract with ethyl acetate (50 mL × 3). Retain the aqueous layer, concentrate to obtain the crude product, and purify by reverse-phase silica gel column chromatography (C18, acetonitrile:water (V / V) = 0:100 - 10:90, gradient elution) to obtain methyl 3-(dimethylphosphoryl)benzoate (2.20 g, yield 88.7%).

[0280] LC-MS, M / Z(ESI): 213.4 [M+H] + .

[0281] Step 2: Synthesis of 3-(dimethylphosphoryl)benzoic acid

[0282]

[0283] Dissolve methyl 3-(dimethylphosphoryl)benzoate (1.00 g, 4.71 mmol) in water (5 mL) and methanol (5 mL), and then add lithium hydroxide (1.98 g, 47.1 mmol). The reaction mixture 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 × 3), and the aqueous layer was retained and concentrated to obtain the crude product. The crude product was purified by reverse-phase chromatography column C18 (acetonitrile: water (V / V) = 0:100 - 10:90, gradient elution) to obtain 3-(dimethylphosphoryl)benzoic acid (600 mg, yield 64.3%).

[0284] LC-MS, M / Z(ESI): 199.2 [M+H] + .

[0285] Step 3: Synthesis of N-((1R,3S)-3-((6-chloro-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)-3-(dimethylphosphoryl)benzamide (Compound 36)

[0286]

[0287] Dissolve 3-(dimethylphosphoryl)benzoic acid (170 mg, 0.86 mmol) in dimethyl sulfoxide (2 mL), and successively add (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), and the reaction mixture was stirred at 25 °C for 18 h under N2 protection. The resulting mixture was diluted with saturated brine (10 mL), then extracted with ethyl acetate (10 mL × 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the 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, yield 42.5%).

[0288] 1 H NMR (400 MHz, CD3OD): δ 8.39 (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).

[0289] LC-MS, M / Z (ESI): 524.2 [M+H] + .

[0290] Example 8: Preparation of N-((1R,3S)-3-((6-chloro-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)-3-(dimethylphosphoryl)benzamide (Compound 38)

[0291] The synthetic route of Compound 38 is shown below:

[0292]

[0293] Step 1: Synthesis of ethyl 3-iodo-1-(2,2,2-trifluoroethyl)-1H-pyrazole-4-carboxylate

[0294]

[0295] At 0 °C, sodium hydride (0.45 g, 11.3 mmol, 60% Wt) was added to a solution of ethyl 3-iodo-1H-pyrazole-4-carboxylate (2.00 g, 7.52 mmol) in tetrahydrofuran (20 mL). The mixture was stirred at 0 °C for 30 minutes, and then 2,2,2-trifluoroethyl trifluoromethanesulfonate (2.62 g, 11.3 mmol) was added. The reaction mixture was stirred at 25 °C for 30 minutes under N2 protection. LC-MS detected the completion of the reaction. The reaction mixture was diluted with water (50 mL), and then extracted with ethyl acetate (50 mL × 3). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was separated and purified by silica gel column chromatography (petroleum ether:ethyl acetate (V / V) = 100:0 - 80:20, gradient elution) to obtain ethyl 3-iodo-1-(2,2,2-trifluoroethyl)-1H-pyrazole-4-carboxylate (370 mg, yield 12.7%).

[0296] LC-MS, M / Z(ESI): 349.2[M+H] + .

[0297] Step 2: Synthesis of ethyl 3-(dimethylphosphoryl)-1-(2,2,2-trifluoroethyl)-1H-pyrazole-4-carboxylate

[0298]

[0299] Ethyl 3-iodo-1-(2,2,2-trifluoroethyl)pyrazole-4-carboxylate (320 mg, 0.92 mmol) was dissolved in dimethyl sulfoxide (2 mL). Under nitrogen protection, 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 in sequence. After the reaction mixture was stirred at 25 °C for 10 minutes, a solution of dimethylphosphine oxide (108 mg, 1.38 mmol) in dimethyl sulfoxide (2 mL) was added. The resulting mixture was stirred at 130 °C for 2 hours under N2 protection. LC-MS detected the completion of the reaction. The reaction mixture was diluted with water (10 mL), and then 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 by C18 column chromatography (water:acetonitrile (V / V) = 100:0 - 90:10, gradient elution) to obtain ethyl 3-(dimethylphosphoryl)-1-(2,2,2-trifluoroethyl)-1H-pyrazole-4-carboxylate (100 mg, yield 35.2%).

[0300] LC-MS, M / Z(ESI): 299.2[M+H] + .

[0301] Step 3: Synthesis of 3-(Dimethylphosphoryl)-1-(2,2,2-trifluoroethyl)-1H-pyrazole-4-carboxylic acid

[0302]

[0303] 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 then lithium hydroxide (141 mg, 3.35 mmol) was added. The resulting mixture was stirred at 25 °C for 1 h. After the reaction was completed as detected by LC-MS, the reaction solution was adjusted to pH = 4 with hydrochloric acid (2 mol / L), extracted with ethyl acetate (10 mL × 3), and the combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated to obtain 3-(dimethylphosphoryl)-1-(2,2,2-trifluoroethyl)-1H-pyrazole-4-carboxylic acid (70.0 mg, yield 77.3%). This crude product was directly used in the next step without further purification.

[0304] LC-MS, M / Z(ESI): 271.1[M+H] + .

[0305] 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

[0306]

[0307] 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 was added successively 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. LC-MS detected the completion of the reaction. 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 obtain the crude product. The crude product was purified by preparative liquid chromatography (Xtimate C18, 21.2*250 mm, 5 μm; 0.1% TFA-ACN; 38 - 68; 20 mL / min) to obtain 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%).

[0308] LC-MS, M / Z(ESI): 596.2[M+H] + .

[0309] 1 H NMR(400 MHz, 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).

[0310] Example 9: Preparation of N-((1R,3S)-3-((6-chloro-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)-3-(dimethylphosphoryl)benzamide (Compound 40)

[0311] The synthetic route of Compound 40 is as follows:

[0312]

[0313] First step: Synthesis of ethyl 3-iodo-1-methyl-1H-pyrazole-4-carboxylate

[0314]

[0315] 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 for 1 h under N2 protection. The formation of the target product was detected by LC-MS. The reaction mixture was diluted with water (100 mL), then extracted with ethyl acetate (50 mL×3). The organic phases were combined, washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered and concentrated to obtain the crude product. The crude product was separated and purified by silica gel column chromatography (petroleum ether:ethyl acetate (V / V)=100:0 - 80:20, gradient elution) to give ethyl 3-iodo-1-methyl-1H-pyrazole-4-carboxylate (900 mg, yield 41.9%).

[0316] LC-MS, M / Z(ESI): 281.1[M+H] + .

[0317] Step 2: Synthesis of ethyl 3-(dimethylphosphoryl)-1-methyl-1H-pyrazole-4-carboxylate

[0318]

[0319] Ethyl 3-iodo-1-methyl-1H-pyrazole-4-carboxylate (600 mg, 2.14 mmol) was dissolved in dimethyl sulfoxide (5 mL). Under nitrogen protection, 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 in sequence. The reaction mixture was stirred at 25 °C for 10 min, then a solution of dimethylphosphine oxide (251 mg, 3.21 mmol) in dimethyl sulfoxide (1 mL) was added. The resulting mixture was stirred at 135 °C for 1 h under N2 protection. The completion of the reaction was detected by LC-MS. The reaction mixture was diluted with water (10 mL), then 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 by reverse-phase C18 column chromatography (water:acetonitrile (V / V)=100:0 - 90:10, gradient elution) to give ethyl 3-(dimethylphosphoryl)-1-methyl-1H-pyrazole-4-carboxylate (250 mg, yield 50.7%).

[0320] LC-MS, M / Z(ESI): 231.2[M+H] + .

[0321] Step 3: Synthesis of 3-(dimethylphosphoryl)-1-methyl-1H-pyrazole-4-carboxylic acid

[0322]

[0323] 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). The resulting mixture was stirred at 25 °C for 1 hour. The reaction was monitored by LC-MS and when completed, methanol was removed by concentration under reduced pressure. The resulting reaction mixture was adjusted to pH = 5 with hydrochloric acid (2 mol / L), and a solid precipitated. The filter cake was collected by filtration and purified by reverse-phase C18 column chromatography (water:acetonitrile (V / V) = 100:0 - 60:40, gradient elution) to give 3-(dimethylphosphoryl)-1-methyl-1H-pyrazole-4-carboxylic acid (120 mg, yield 68.3%).

[0324] LC-MS, M / Z(ESI): 203.0 [M-H] + .

[0325] Step 4: Synthesis of N-((1R,3S)-3-((6-chloro-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)-3-(dimethylphosphoryl)benzamide (Compound 40)

[0326]

[0327] 3-(Dimethylphosphoryl)-1-methyl-1H-pyrazole-4-carboxylic acid (100 mg, 0.49 mmol) was dissolved in DMSO (2 mL), and successively (1S,3R)-N 1 -(6-chloro-2-(trifluoromethyl)quinolin-4-yl)cyclohexane-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), and 1-hydroxybenzotriazole (201 mg, 1.48 mmol) were added. The resulting mixture was stirred at 25 °C for 18 hours under N2 protection. The reaction was monitored by LC-MS and when completed, 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 the crude product. The crude product was purified by normal-phase silica gel column chromatography (petroleum ether:ethyl acetate (V / V) = 100:1 - 0:100, gradient elution) to give N-((1R,3S)-3-((6-chloro-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)-3-(dimethylphosphoryl)benzamide (Compound 40) (47.4 mg, yield 17.9%).

[0328] 1 1H NMR (400 MHz, CD3OD): δ 8.38 (d, 1H), 8.20 (s, 1H), 7.92 (d, 1H), 7.70 (dd, J = 9.2, 2.4 Hz 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).

[0329] LC-MS, M / Z (ESI): 528.2 [M + H] + .

[0330] 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)

[0331] The synthetic route of Compound 41 is as follows:

[0332]

[0333] The first step: Synthesis of methyl 3 - formyl - 1 - methyl - 1H - pyrazole - 4 - carboxylate

[0334]

[0335] 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), then [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium (484 mg, 0.662 mmol) was added. After addition, the reaction mixture was stirred at 60 °C for 12 hours under a carbon monoxide (15 psi) atmosphere. After the reaction was completed, the reaction mixture was cooled to room temperature, filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by column chromatography (PE:EA (V / V) = 10:1 to 1:1) to obtain methyl 3 - formyl - 1 - methyl - 1H - pyrazole - 4 - carboxylate (1.10 g, yield 49.5%).

[0336] LC / MS (ESI) (m / z): 169.0 (M + H) + .

[0337] Step 2: Synthesis of methyl 3-(hydroxymethyl)-1-methyl-1H-pyrazole-4-carboxylate

[0338]

[0339] At room temperature, methyl 3-formyl-1-methyl-1H-pyrazole-4-carboxylate (1.10 g, 6.54 mmol) was dissolved in methanol (20 mL), and then sodium borohydride (272 mg, 7.20 mmol) was added portionwise under an ice bath. The reaction mixture was stirred under an ice bath for 2 hours. After completion of the reaction, 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, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The obtained crude product was purified by column chromatography (PE:EA (V / V) = 10:1 to 0:1) to give methyl 3-(hydroxymethyl)-1-methyl-1H-pyrazole-4-carboxylate (800 mg, yield 71.9%).

[0340] LC / MS(ESI)(m / z):171.0(M+H) + .

[0341] Step 3: Synthesis of methyl 3-((dimethylcarbamoyloxy)methyl)-1-methyl-1H-pyrazole-4-carboxylate

[0342]

[0343] At room temperature, sodium hydride (85.0 mg, 2.12 mmol) was dissolved in tetrahydrofuran (3 mL), and then methyl 3-(hydroxymethyl)-1-methyl-1H-pyrazole-4-carboxylate (300 mg, 1.76 mmol) was slowly added at 0 °C. The reaction mixture was stirred at room temperature for 1 hour, and then dimethylcarbamoyl chloride (284 mg, 2.64 mmol) was added dropwise. After addition, the temperature was raised to 80 °C and stirred for 1 hour. The reaction mixture was cooled to room temperature, quenched with saturated ammonium chloride (2 mL) under an ice bath, and then extracted with dichloromethane (5 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered and concentrated to obtain the crude product methyl 3-((dimethylcarbamoyloxy)methyl)-1-methyl-1H-pyrazole-4-carboxylate (424 mg, 1.76 mmol, yield 100%), which was directly used for the next step reaction.

[0344] LC / MS(ESI)(m / z):242.1(M+H) + .

[0345] Step 4: Synthesis of 3-((dimethylcarbamoyloxy)methyl)-1-methyl-1H-pyrazole-4-carboxylic acid

[0346]

[0347] To a solution of methyl 3-((dimethylcarbamoyloxy)methyl)-1-methyl-1H-pyrazole-4-carboxylate (424 mg, 1.76 mmol, yield 100%) in tetrahydrofuran (3 mL) was added dropwise a solution of lithium hydroxide (127 mg, 5.29 mmol) in water (3 mL) at 0 °C. The reaction mixture was stirred at room temperature for 12 h. After completion of the reaction, the pH of the reaction mixture was adjusted to 3 with 1N HCl solution under ice bath conditions, and then extracted with ethyl acetate (3 × 10 mL). The organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered and concentrated to obtain 3-((dimethylcarbamoyloxy)methyl)-1-methyl-1H-pyrazole-4-carboxylic acid (300 mg, yield 75%).

[0348] LC / MS(ESI)(m / z): 228.0(M+H) +

[0349] 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)

[0350]

[0351] At room temperature, (1S,3R)-N 1 -(6-chloro-2-(trifluoromethyl)quinolin-4-yl)cyclohexane-1,3-diamine (Intermediate A1) (45.0 mg, 0.130 mmol) and 3-((dimethylcarbamoyloxy)methyl)-1-methyl-1H-pyrazole-4-carboxylic acid (30.0 mg, 0.130 mmol) were dissolved in N,N-dimethylformamide (0.5 mL), then diisopropylethylamine (51.6 mg, 0.400 mmol) and 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (49.0 mg, 0.130 mmol) were added. The reaction mixture was stirred at 25 °C for 2 h. After completion of the reaction, the reaction mixture was filtered, and the filtrate was concentrated to dryness. The obtained crude product was purified by preparative liquid chromatography (column: YMC-Triart Prep C18 7μm 30mm×40cm; solvent: A = 0.1% ammonia water, B = acetonitrile; gradient: 30%-70%, 9 min) to obtain (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, yield 20.7%).

[0352] LC / MS(ESI)(m / z): 553.10 (M+H) + ;

[0353] 1 H NMR(400 MHz, DMSO-d6) δ 8.54 (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).

[0354] Example 11: Preparation of 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)

[0355] The synthetic route of Compound 42 is as follows:

[0356]

[0357] First step: Synthesis of ethyl 1-(2-hydroxy-2-methylpropyl)-3-iodo-1H-pyrazole-4-carboxylate

[0358]

[0359] 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). The resulting mixture was stirred at 25 °C for 1 hour under N2 protection. LC-MS was used to detect the completion of the reaction. The reaction solution was diluted with water (100 mL), then extracted with ethyl acetate (100 mL × 3). The organic layers were combined, washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered and concentrated to obtain the crude product. The crude product was separated and purified by normal-phase silica gel column (petroleum ether:ethyl acetate (V / V) = 100:0 - 80:20, gradient elution) to obtain ethyl 1-(2-hydroxy-2-methylpropyl)-3-iodo-1H-pyrazole-4-carboxylate (900 mg, yield 41.9%).

[0360] Step 2: Synthesis of Ethyl 3-((dimethyl(oxo)-λ 6 -aminosulfonylimide)-1-(2-hydroxy-2-methylpropyl)-1H-pyrazole-4-carboxylate

[0361]

[0362] 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 successively added dimethylsulfoximine (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). The resulting mixture was stirred at 110 °C for 18 h. After completion of the reaction detected by LC-MS, the reaction solution was diluted with water (50 mL), then extracted with ethyl acetate (50 mL × 3). The combined organic layers were washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was separated and purified by a normal-phase silica gel column (dichloromethane:methanol (V / V) = 100:0 - 90:10, gradient elution) to give ethyl 3-((dimethyl(oxo)-λ 6 -aminosulfonylimide)-1-(2-hydroxy-2-methylpropyl)-1H-pyrazole-4-carboxylate (480 mg, yield 68.8%).

[0363] LC-MS, M / Z (ESI): 304.2 [M+H] + .

[0364] Step 3: Synthesis of 3-((dimethyl(oxo)-λ 6 -sulfanidylpyridyl)amino)-1-(2-hydroxy-2-methylpropyl)-1H-pyrazole-4-carboxylic acid

[0365]

[0366] To 3-((dimethyl(oxo)-λ 6To a solution of ethyl 3-((dimethyl(oxo)-λ6-sulfanylideneamino)sulfonyl)-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). The resulting mixture was stirred at 20 °C for 18 h. LC-MS detected the completion of the reaction. The reaction solution was concentrated under reduced pressure to remove methanol. The resulting mixture was adjusted to pH 5 with hydrochloric acid (2 mol / L), and the precipitate was collected by filtration and dried to obtain 3-((dimethyl(oxo)-λ6-sulfanylideneamino)sulfonyl)-1-(2-hydroxy-2-methylpropyl)-1H-pyrazole-4-carboxylic acid (180 mg, yield 89.3%). The crude product was used directly in the next step of synthesis without further purification.

[0367] LC-MS, M / Z (ESI): 276.2 [M-H] - .

[0368] Step 4: Synthesis of N-((1R,3S)-3-((6-chloro-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)-3-(dimethyl(oxo)-λ 6 6-sulfanylideneamino)sulfonyl)-1-(2-hydroxy-2-methylpropyl)-1H-pyrazole-4-carboxamide

[0369]

[0370] Dissolve 3-((dimethyl(oxo)-λ 6 6-sulfanylideneamino)sulfonyl)-1-(2-hydroxy-2-methylpropyl)-1H-pyrazole-4-carboxylic acid (100 mg, 0.36 mmol) in dimethyl sulfoxide (2 mL). Sequentially add (1S,3R)-N 1 -(6-chloro-2-(trifluoromethyl)quinolin-4-yl)cyclohexane-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 h under N2 protection. LC-MS detected the completion of the reaction. The reaction solution was diluted with saturated brine (20 mL), then extracted with ethyl acetate (20 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 by silica gel column chromatography (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)-λ6 -Aminosulfonamido)-1-(2-hydroxy-2-methylpropyl)-1H-pyrazole-4-carboxamide (Compound 42) (109 mg, yield 48.6%).

[0371] 1 H NMR (400 MHz, DMSO-d6): δ 8.58 (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).

[0372] LC-MS, M / Z (ESI): 601.2 [M+H] + .

[0373] 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)

[0374] The synthetic route of Compound 43 is as follows:

[0375]

[0376] First step: Synthesis of ethyl 3-chloro-1-(2-hydroxyethyl)-1H-pyrazole-4-carboxylate

[0377]

[0378] Ethyl 3-chloro-1H-pyrazole-4-carboxylate (0.500 g, 2.86 mmol) and potassium carbonate (0.792 g, 5.73 mmol) were added to acetonitrile (15 mL), then 2-bromoethanol (394 mg, 3.15 mmol) was added, and the reaction mixture was heated to 60 °C and stirred for 12 hours. After completion of the reaction, the reaction mixture was filtered, the filtrate was concentrated, and the obtained crude product was purified by column chromatography (PE:EA (V / V) = 10:1 to 1:1) to obtain ethyl 3-chloro-1-(2-hydroxyethyl)-1H-pyrazole-4-carboxylate (0.6 g, yield 96.0%).

[0379] LC / MS (ESI) (m / z): 219.0 (M+H)+ .

[0380] Step 2: Synthesis of Ethyl 3-chloro-1-(2-((dimethylcarbamoyl)oxy)ethyl)-1H-pyrazole-4-carboxylate

[0381]

[0382] Sodium hydride (66.0 mg, 1.65 mmol, 60%) was dissolved in tetrahydrofuran (3 mL), and ethyl 3-chloro-1-(2-hydroxyethyl)-1H-pyrazole-4-carboxylate (300 mg, 1.37 mmol) was slowly added thereto under an ice bath. The reaction solution was stirred at room temperature for 1 hour, and then dimethylcarbamoyl chloride (221 mg, 2.06 mmol) was added dropwise thereto under an ice bath. After the addition, the reaction solution was stirred at room temperature for 12 hours. The reaction solution was cooled to room temperature, quenched with saturated ammonium chloride (2 mL) under an ice bath, and then extracted with dichloromethane (5 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered and concentrated to obtain the crude product ethyl 3-chloro-1-(2-((dimethylcarbamoyl)oxy)ethyl)-1H-pyrazole-4-carboxylate (397 mg, 1.37 mmol, yield 100%), which was directly used in the next step of the reaction.

[0383] LC / MS(ESI)(m / z):290.1(M+H) + .

[0384] Step 3: Synthesis of 3-chloro-1-(2-((dimethylcarbamoyl)oxy)ethyl)-1H-pyrazole-4-carboxylic acid

[0385]

[0386] A solution of lithium hydroxide (164 mg, 6.83 mmol) in water (3 mL) was added dropwise 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) at 0 °C. The reaction solution was stirred at room temperature for 12 hours. After the reaction was completed, the reaction solution was adjusted to pH 3 with 4N HCl solution under an ice bath condition, and then extracted with ethyl acetate (3 × 10 mL). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to obtain the crude product 3-chloro-1-(2-((dimethylcarbamoyl)oxy)ethyl)-1H-pyrazole-4-carboxylic acid (300 mg, yield 83%).

[0387] LC / MS(ESI)(m / z):262.0(M+H) +

[0388] 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)

[0389]

[0390] Dissolve (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-((dimethylcarbamoyl)oxy)ethyl)-1H-pyrazole-4-carboxylic acid (57.0 mg, 0.220 mmol) in N,N-dimethylformamide (0.5 mL), then add diisopropylethylamine (56.0 mg, 0.440 mmol) and 2-(7-azabenzotriazol)-N,N,N',N'-tetramethyluronium hexafluorophosphate (84.0 mg, 0.220 mmol), and stir the reaction mixture at 25 °C for 2 h. After completion of the reaction, filter the reaction mixture, concentrate the filtrate under reduced pressure, and purify the obtained crude product by preparative high performance liquid chromatography (column: YMC-Triart Prep C18 7 μm 30 mm × 40 cm; solvent: A = 0.1% ammonia water, B = acetonitrile; gradient: 30% - 82%, 9 min) to obtain 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%).

[0391] LC / MS(ESI)(m / z): 587.10(M+H) + ;

[0392] 1 1H NMR(400 MHz, DMSO-d6) δ 8.55(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).

[0393] 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)

[0394] The synthetic route of Compound 44 is as follows:

[0395]

[0396] First step: Synthesis of ethyl 3-chloro-1-((methylthio)methyl)-1H-pyrazole-4-carboxylate

[0397]

[0398] At room temperature, dissolve ethyl 3-chloro-1H-pyrazole-4-carboxylate (2.00 g, 11.46 mmol) in anhydrous tetrahydrofuran (20 mL), stir in an ice bath at 0 °C for 10 min, then add sodium hydride (504 mg, 12.60 mmol) in portions, and then slowly add chloromethyl methyl sulfide (1.33 g, 13.75 mmol). After the reaction solution slowly rises to room temperature, continue to stir for 30 min. After the reaction is completed, stop stirring, pour the reaction solution into water (30 mL), then extract with ethyl acetate (30 mL × 3), collect the organic phase and dry it with anhydrous sodium sulfate, filter and concentrate. The residue is 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, yield 34.59%).

[0399] LC-MS, M / Z (ESI): 235.3 [M+H] +

[0400] 1 1H NMR (400 MHz, DMSO-d6) δ 8.50 (s, 1H), 5.26 (s, 2H), 4.23 (q, 2H), 2.14 (s, 3H), 1.27 (t, 3H).

[0401] Second step: Synthesis of ethyl 3-chloro-1-((methylsulfonyl)methyl)-1H-pyrazole-4-carboxylate

[0402]

[0403] At room temperature, dissolve ethyl 3-chloro-1-((methylthio)methyl)-1H-pyrazole-4-carboxylate (550 mg, 2.34 mmol) in anhydrous dichloromethane (11 mL), stir at 0 °C for 10 min, then add m-chloroperoxybenzoic acid (1.21 g, 7.03 mmol). The reaction mixture is slowly warmed to room temperature and stirred for an additional 2 h. After thin-layer chromatography shows that the starting material has reacted completely, stop stirring, pour the reaction mixture into saturated aqueous sodium thiosulfate (30 mL), then extract with ethyl acetate (30 mL × 3). Collect the organic phase and wash it with saturated aqueous sodium carbonate (40 mL × 3), then dry over anhydrous sodium sulfate, filter and concentrate to obtain ethyl 3-chloro-1-((methylsulfonyl)methyl)-1H-pyrazole-4-carboxylate (615 mg, yield 98.40%). The crude product obtained can be directly used in the next reaction.

[0404] LC-MS, M / Z(ESI): 267.0[M+H] +

[0405] Step 3: Synthesis of ethyl 3-chloro-1-(1-(methylsulfonyl)ethyl)-1H-pyrazole-4-carboxylate

[0406]

[0407] At room temperature, dissolve ethyl 3-chloro-1-((methylsulfonyl)methyl)-1H-pyrazole-4-carboxylate (200 mg, 0.75 mmol) in anhydrous DMF (2 mL), then add cesium carbonate (488.67 mg, 1.50 mmol). After stirring the reaction mixture for 10 min, slowly add methyl iodide (212.88 mg, 1.50 mmol), and then continue to stir at room temperature for 12 h. After TLC monitoring shows that the starting material has reacted completely, stop stirring, add water (5 mL) to quench the reaction, then extract with ethyl acetate (15 mL × 3). Collect the organic phase and dry over anhydrous sodium sulfate, filter and concentrate. The residue is separated and purified by silica gel column chromatography (DCM:MeOH (V / V)=100:2) to obtain ethyl 3-chloro-1-(1-(methylsulfonyl)ethyl)-1H-pyrazole-4-carboxylate (139 mg, yield 66.03%).

[0408] LC-MS, M / Z(ESI): 281.0[M+H] +

[0409] Step 4: Synthesis of 3-chloro-1-(1-(methylsulfonyl)ethyl)-1H-pyrazole-4-carboxylic acid

[0410]

[0411] At room temperature, ethyl 3-chloro-1-(1-(methylsulfonyl)ethyl)-1H-pyrazole-4-carboxylate (130 mg, 0.46 mmol) was dissolved in a mixed solution of methanol (2 mL) and water (0.8 mL), and then sodium hydroxide (148 mg, 3.70 mmol) was added. The reaction mixture was stirred at room temperature for 2 h. After monitoring by TLC showed that the raw material had reacted completely, the stirring was stopped, and methanol was removed by distillation under reduced pressure. Water (5 mL) was added to the residue for dilution, and then the pH was adjusted to 2 with dilute hydrochloric acid (2 mol / L). The mixture was extracted with ethyl acetate (15 mL × 3), the organic phase was collected and dried over anhydrous sodium sulfate, and the organic phase was concentrated by distillation under reduced pressure to obtain ethyl 3-chloro-1-(1-(methylsulfonyl)ethyl)-1H-pyrazole-4-carboxylate (110 mg, yield 94.01%). The obtained crude product could be directly used for the next reaction.

[0412] LC-MS, M / Z(ESI): 252.8[M+H] +

[0413] 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)

[0414]

[0415] At room temperature, 3-chloro-1-(1-(methylsulfonyl)ethyl)-1H-pyrazole-4-carboxylic acid (1119E) (50 mg, 0.20 mmol) was dissolved in anhydrous DMF (1 mL), stirred at 0 °C for 10 min, then DIPEA (102 mg, 0.79 mmol) and HATU (151 mg, 0.40 mmol) were added, and 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), the reaction solution was slowly warmed to room temperature and stirred for an additional 6 h. After monitoring by TLC showed that the raw materials had reacted completely, stirring was stopped, water (5 mL) was added to quench the reaction, and then extraction was carried out 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 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, yield 48.05%).

[0416] LC-MS, M / Z (ESI): 578.2 [M+H] +

[0417] 1 1H NMR (400 MHz, DMSO-d6) δ 8.59 (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).

[0418] Example 14: Preparation of N-((1R,3S))-3-((6-chloro-2-trifluoromethyl)(quinolin-4-yl)amino)cyclohexyl)-3-((methylsulfonyl)methyl)benzamide (Compound 45)

[0419] The synthetic route of Compound 45 is shown below:

[0420]

[0421] 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-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (133 mg, 0.350 mmol) was added. After the addition, the reaction mixture was stirred at room temperature for 12 h. After completion of the reaction, the reaction mixture was concentrated under reduced pressure. The resulting crude product was purified by preparative high performance liquid chromatography (column: YMC-Triart Prep C18 7 μm 30 mm×40 cm; solvent: A = 0.1% aqueous ammonia, B = acetonitrile; gradient: 30%-72%, 9 min) to give the compound N-((1R,3S))-3-((6-chloro-2-(trifluoromethyl))(quinolin-4-yl)amino)cyclohexyl)-3-((methylsulfonyl)methyl)benzamide (Compound 44) (62.0 mg, yield 49.9%).

[0422] LC / MS(ESI)(m / z):540.10(M+H) + ;

[0423] 1 1H 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).

[0424] 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)

[0425] The synthetic route of Compound 46 is shown below:

[0426]

[0427] First step: Synthesis of ethyl 3-chloro-1-(1-(methylsulfonyl)ethyl)-1H-pyrazole-4-carboxylate

[0428]

[0429] 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 hydride (60%, 40.0 mg, 0.980 mmol) was slowly added under an ice bath. The reaction mixture was stirred for 20 minutes under the ice bath, and then iodomethane (140 mg, 0.980 mmol) was slowly added. After the addition, the reaction mixture was warmed to 25 °C and stirred for 12 hours. After completion of the reaction, the reaction was quenched by dropwise addition of saturated ammonium chloride solution (2 mL) under an ice bath. The mixture was extracted with ethyl acetate (5 mL × 3), and the organic phases were combined, dried over anhydrous sodium sulfate, filtered and concentrated to obtain ethyl 3-chloro-1-(2-(methylsulfonyl)propan-2-yl)-1H-pyrazole-4-carboxylate (46-2) (0.140 g, yield 97.1%).

[0430] LC / MS(ESI)(m / z):295.0(M+H) + .

[0431] Step 2: Synthesis of 3-chloro-1-(1-(methylsulfonyl)ethyl)-1H-pyrazole-4-carboxylic acid

[0432]

[0433] 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), and then a solution of lithium hydroxide (100 mg, 2.38 mmol) in water (1 mL) was slowly added. After the addition, 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 an ice bath, and then the mixture was 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 obtain 3-chloro-1-(1-(methylsulfonyl)ethyl)-1H-pyrazole-4-carboxylic acid (46-3) (126 mg, yield 100%).

[0434] LC / MS(ESI)(m / z):267.0(M+H) +

[0435] 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)

[0436]

[0437] 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), then diisopropylethylamine (61.0 mg, 0.470 mmol) and 2-(7-azabenzotriazol)-N,N,N',N'-tetramethyluronium hexafluorophosphate (110 mg, 0.280 mmol) were added, and the reaction mixture was stirred at 25 °C for 12 hours. After completion of the reaction, the reaction mixture was filtered, the filtrate was concentrated, and the obtained crude product was purified by preparative high performance liquid chromatography (column: YMC-Actus Triart C18 ExRS-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 obtain 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%).

[0438] LC / MS (ESI) (m / z): 592.0 (M+H) + ;

[0439] 1 1H NMR (400 MHz, DMSO-d6) δ 8.61 (s, 1H), 8.56 (s, 1H), 7.96 (d, 1H), 7.86 (d, 1H), 7.71 (d, 1H), 7.44 (d, 1H), 6.91 (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).

[0440] Example 16: Preparation of 3-chloro-N-((1R,3S))-3-((6-chloro-2-trifluoromethyl))quinolin-4-yl)amino)cyclohexyl)-1-((S-methylsulfinimidoyl)methyl)-1H-pyrazole-4-carboxamide (Compound 47)

[0441] The synthetic route of Compound 47 is as follows:

[0442]

[0443] Step 1: Synthesis of Ethyl 3-chloro-1-((S-methylsulfinylimino)methyl)-1H-pyrazole-4-carboxylate

[0444]

[0445] At room temperature, dissolve ethyl 3-chloro-1-((methylthio)methyl)-1H-pyrazole-4-carboxylate (44-2) (400 mg, 1.70 mmol) in anhydrous methanol (15 mL), then successively add iodobenzene diacetate (1.70 g, 5.10 mmol) and ammonium carbonate (490 mg, 5.10 mmol). After the addition is complete, stir at room temperature for 4 hours. After monitoring by TLC shows that the raw materials have reacted completely, stop stirring, add water (50 mL) for dilution, then extract with ethyl acetate (30 mL×3). Collect the organic phase and dry it with anhydrous sodium sulfate, filter and concentrate. The residue is separated and purified by silica gel column chromatography (PE:EA = 100:10 (V / V)) to obtain ethyl 3-chloro-1-((S-methylsulfinylimino)methyl)-1H-pyrazole-4-carboxylate (47-1) (376 mg, yield 83.03%).

[0446] LC-MS, M / Z(ESI): 266.0[M+H] + .

[0447] Step 2: Synthesis of 3-chloro-1-((S-methylsulfinylamino)methyl)-1H-pyrazole-4-carboxylic acid (47-2)

[0448]

[0449] At room temperature, dissolve ethyl 3-chloro-1-((S-methylsulfinylimino)methyl)-1H-pyrazole-4-carboxylate (47-1) (200 mg, 0.75 mmol) in a mixed solution of tetrahydrofuran / methanol / water (2 mL / 1 mL / 1 mL), then slowly add lithium hydroxide monohydrate (253 mg, 6.02 mmol) under ice bath conditions. After the addition is complete, stir the reaction solution at room temperature for 2 hours. After monitoring by TLC shows that the raw materials have reacted completely, stop stirring, add water (10 mL) to dilute the reaction solution, adjust the pH to 4 with dilute hydrochloric acid (1 mol / L), then extract with ethyl acetate (30 mL×3). Collect the organic phase and dry it with anhydrous sodium sulfate, filter and concentrate to obtain 3-chloro-1-((S-methylsulfinylamino)methyl)-1H-pyrazole-4-carboxylic acid (47-2) (150 mg, yield 83.86%).

[0450] LC-MS, M / Z (ESI): 238.0 [M+H] + .

[0451] Step 3: Synthesis of 3-chloro-N-((1R,3S))-3-((6-chloro-2-(trifluoromethyl))quinolin-4-yl)amino)cyclohexyl)-1-((S-methylsulfinimidoyl)methyl)-1H-pyrazole-4-carboxamide (Compound 47)

[0452]

[0453] At room temperature, dissolve 3-chloro-1-((S-methylsulfinimidoyl)methyl)-1H-pyrazole-4-carboxylic acid (47-2) (50 mg, 0.21 mmol) in anhydrous N,N-dimethylformamide (1 mL), stir for 10 minutes in an ice bath, then add 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). Continue to stir the reaction mixture in the ice bath for 30 minutes, then add (1S,3R)-N 1 -(6-chloro-2-(trifluoromethyl)quinolin-4-yl)cyclohexane-1,3-diamine (80 mg, 0.23 mmol). After the addition is complete, slowly warm to room temperature and continue to stir for 6 hours. When TLC monitoring shows that the raw materials have reacted completely, stop stirring, add water (5 mL) for dilution, then extract with ethyl acetate (5 mL×3). Collect the organic phase and dry it over anhydrous sodium sulfate. Concentrate the organic phase by vacuum distillation. Purify the residue 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-methylsulfinimidoyl)methyl)-1H-pyrazole-4-carboxamide (Compound 47) (23 mg, yield 19.40%).

[0454] LC-MS, M / Z (ESI): 563.0 [M+H] + .

[0455] 11H NMR (400 MHz, DMSO-d6): δ 8.59 (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).

[0456] Example 17: Preparation of 3-chloro-N-((1R,3S)-3-((6-chloro-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)-1-((N,S-dimethylsulfimidoyl)methyl)-1H-pyrazole-4-carboxamide (Compound 51)

[0457] The synthetic route of Compound 51 is as follows:

[0458]

[0459] First step: Ethyl 3-chloro-1-((N,S-dimethylsulfimidoyl)methyl)-1H-pyrazole-4-carboxylate

[0460]

[0461] At room temperature, dissolve ethyl 3-chloro-1-((S-methylsulfimidoyl)methyl)-1H-pyrazole-4-carboxylate (47-1) (50.0 mg, 0.190 mmol) in N,N-dimethylformamide (1 mL), then add cesium carbonate (61.0 mg, 0.190 mmol) and methyl iodide (32.0 mg, 0.230 mmol) at room temperature. After addition, stir at 25 °C for 12 hours. After the reaction is complete, dilute the reaction solution with ethyl acetate (20 mL), wash successively with water (10 mL) and saturated brine (10 mL), and concentrate the organic phase under reduced pressure to obtain ethyl 3-chloro-1-((N,S-dimethylsulfimidoyl)methyl)-1H-pyrazole-4-carboxylate (51-1) (53.0 mg, yield 100%). LC / MS (ESI) (m / z): 280.0 (M + H) + .

[0462] Second step: 3-chloro-1-((N,S-dimethylsulfimidoyl)methyl)-1H-pyrazole-4-carboxylic acid

[0463]

[0464] At room temperature, ethyl 3-chloro-1-((N,S-dimethylsulfamoylimino)methyl)-1H-pyrazole-4-carboxylate (51-1) (53.0 mg, 0.190 mmol) was dissolved in a mixed solution of tetrahydrofuran (0.4 mL) / methanol (0.2 mL) / water (0.2 mL), and then lithium hydroxide monohydrate (40.0 mg, 0.950 mmol) was added at room temperature. After the addition, the reaction solution was stirred at 25 °C for 12 hours. After the reaction was completed, the pH was adjusted to 3 with dilute hydrochloric acid (1 mol / L) under an ice bath, and then extracted with ethyl acetate (20 mL), washed successively with water (5 mL) and saturated brine (5 mL), and the organic phase was concentrated under reduced pressure to obtain 3-chloro-1-((N,S-dimethylsulfamoylimino)methyl)-1H-pyrazole-4-carboxylic acid (51-2) (48.0 mg, 0.190 mmol, yield 100%). LC / MS (ESI) (m / z): 252.0 (M+H) +

[0465] Step 3: 3-Chloro-N-((1R,3S)-3-((6-chloro-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)-1-((N,S-dimethylsulfamoylimino)methyl)-1H-pyrazole-4-carboxamide (Compound 51)

[0466]

[0467] At room temperature, 3-chloro-1-((N,S-dimethylsulfimidoyl)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-azabenzotriazol-1-yl)-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 successively. The reaction mixture was stirred at 25 °C for 12 h. After completion of the reaction, the reaction mixture was diluted with ethyl acetate (20 mL) and washed successively with water (10 mL) and saturated brine (10 mL). The organic phase was concentrated under reduced pressure, and the crude product obtained was purified by preparative high performance liquid chromatography (column: YMC-Triart Prep C18 7μm 30mm×40cm; solvent: A = 0.1% aqueous ammonia, B = acetonitrile; gradient: 25%-76%, 9 min) to give 3-chloro-N-((1R,3S)-3-((6-chloro-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)-1-((N,S-dimethylsulfimidoyl)methyl)-1H-pyrazole-4-carboxamide (Compound 51) (4.90 mg, yield 4.5%).

[0468] LC / MS(ESI)(m / z):577.0(M+H) + ;

[0469] 1 1H 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).

[0470] Example 18: Preparation of 3-chloro-N-[(1R,3S)-3-((6-chloro-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl]-1-[1-(methyldioxido-λ6-sulfanyl)propyl]pyrazole-4-carboxamide (Compound 56)

[0471] The synthetic route of Compound 56 is shown as follows:

[0472]

[0473] Step 1: Synthesis of Ethyl 3-chloro-1-(1-(methylsulfonyl)propyl)-1H-pyrazole-4-carboxylate

[0474]

[0475] At room temperature, dissolve ethyl 3-chloro-1-((methylsulfonyl)methyl)-1H-pyrazole-4-carboxylate (44-3) (40.0 mg, 0.150 mmol) in N,N-dimethylformamide (0.5 mL) and tetrahydrofuran (0.5 mL). Add sodium hydride (6.0 mg, 0.150 mmol, 60%) under an ice bath. After 20 minutes, slowly add iodoethane (23.4 mg, 0.150 mmol). The reaction mixture is slowly warmed to 25 °C and stirred for 12 hours. After completion of the reaction, quench with saturated ammonium chloride solution (10 mL) under an ice bath, then extract with ethyl acetate (20 mL). The organic phase is washed successively with water (10 mL) and saturated brine (10 mL). The organic phase is concentrated under reduced pressure to obtain ethyl 3-chloro-1-(1-(methylsulfonyl)propyl)-1H-pyrazole-4-carboxylate (56-1) (44.0 mg, yield 100%).

[0476] LC / MS(ESI)(m / z): 295.0 (M+H) + .

[0477] Step 2: 3-chloro-1-(1-(methylsulfonyl)propyl)-1H-pyrazole-4-carboxylic acid

[0478]

[0479] At room temperature, dissolve ethyl 3-chloro-1-(1-(methylsulfonyl)propyl)-1H-pyrazole-4-carboxylate (56-1) (44.0 mg, 0.150 mmol) in tetrahydrofuran (0.4 mL) / methanol (0.2 mL) / water (0.2 mL). Then add lithium hydroxide monohydrate (31.0 mg, 0.750 mmol) at room temperature. After addition, the reaction mixture is stirred at 25 °C for 12 hours. After completion of the reaction, adjust the pH to 3 with dilute hydrochloric acid (1 mol / L) under an ice bath, then extract with ethyl acetate (20 mL). Wash successively with water (5 mL) and saturated brine (5 mL). The organic phase is concentrated under reduced pressure to obtain 3-chloro-1-(1-(methylsulfonyl)propyl)-1H-pyrazole-4-carboxylic acid (56-2) (40.0 mg, yield 100%).

[0480] LC / MS(ESI)(m / z): 267.0 (M+H) +

[0481] 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

[0482]

[0483] At room temperature, dissolve 3-chloro-1-(1-(methylsulfonyl)propyl)-1H-pyrazole-4-carboxylic acid (56-2) (40.0 mg, 0.150 mmol) in N,N-dimethylformamide (1.0 mL), then successively add diisopropylethylamine (0.052 mL, 0.300 mmol), (7-azabenzotriazol-1-yl)-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), and stir the reaction solution at 25 °C for 12 hours. After the reaction is completed, dilute the reaction solution with ethyl acetate (20 mL), wash it successively with water (10 mL) and saturated brine (10 mL), concentrate the organic phase under reduced pressure, and purify the obtained crude product by preparative high performance liquid chromatography (column: YMC-Actus Triart C 18 ExRS-5μm 100*30mm; solvent: A = 0.1% FA, B = acetonitrile; gradient: 30-80% 9 min), 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%).

[0484] LC / MS(ESI)(m / z):592.0(M+H) + ;

[0485] 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,1H),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).

[0486] 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)

[0487] The synthetic route of Compound 64 is as follows:

[0488]

[0489] Step 1: Synthesis of ethyl 3-chloro-1-hydroxymethyl-1H-pyrazole-4-carboxylate

[0490]

[0491] 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). The reaction mixture was stirred at 25 °C for 18 hours. After completion of the reaction, the reaction mixture was quenched with saturated brine (100 mL), then extracted with ethyl acetate (100 mL × 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate (V / V) = 100:0 - 50:50) to obtain ethyl 3-chloro-1-hydroxymethyl-1H-pyrazole-4-carboxylate (1.04 g, yield 88.7%).

[0492] LC-MS, M / Z (ESI): 205.0 [M+H] +

[0493] Step 2: Synthesis of ethyl 1-bromomethyl-3-chloro-1H-pyrazole-4-carboxylate

[0494]

[0495] To a solution of ethyl 3-chloro-1-hydroxymethyl-1H-pyrazole-4-carboxylate (804 mg, 3.93 mmol) in diethyl ether (10 mL) was added phosphorus tribromide (2.13 g, 7.86 mmol). The reaction mixture was stirred at 15 °C for 18 hours. After completion of the reaction, the reaction mixture was quenched with water (100 mL), then extracted with ethyl acetate (100 mL × 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate (V / V) = 100:0 to 50:50) to obtain ethyl 1-bromomethyl-3-chloro-1H-pyrazole-4-carboxylate (760 mg, yield 72.3%).

[0496] LC-MS, M / Z (ESI): 267.0 [M+H] +

[0497] Step 3: Synthesis of Ethyl 3-chloro-1-((ethylthio)methyl)-1H-pyrazole-4-carboxylate

[0498]

[0499] 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 ethylthiolate (717 mg, 8.52 mmol), and the reaction mixture was stirred at 25 °C for 18 h. After completion of the reaction, the reaction mixture was quenched with water (100 mL), then extracted with ethyl acetate (150 mL × 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was purified by silica gel column chromatography (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, yield 93.0%).

[0500] LC-MS, M / Z(ESI): 249.0 [M+H] +

[0501] Step 4: Synthesis of Ethyl 3-chloro-1-((ethylsulfonyl)methyl)-1H-pyrazole-4-carboxylate

[0502]

[0503] Compound ethyl 3-chloro-1-((ethylthio)methyl)-1H-pyrazole-4-carboxylate (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 mixture was stirred at 25 °C for 2 h. After completion of the reaction, the reaction mixture was poured into saturated aqueous sodium thiosulfate (50 mL), then extracted with ethyl acetate (100 mL × 3). The organic phase was collected and washed with saturated aqueous sodium carbonate (50 mL × 3), then dried over anhydrous sodium sulfate, filtered and concentrated to give ethyl 3-chloro-1-((ethylsulfonyl)methyl)-1H-pyrazole-4-carboxylate (670 mg, yield 75.4%).

[0504] LC-MS, M / Z(ESI): 281.0 [M+H] +

[0505] Step 5: Synthesis of Ethyl 3-chloro-1-(1-(ethylsulfonyl)ethyl)-1H-pyrazole-4-carboxylate

[0506]

[0507] Dissolve ethyl 3-chloro-1-((ethylsulfonyl)methyl)-1H-pyrazole-4-carboxylate (670 mg, 2.39 mmol) in anhydrous DMF (6 mL), add cesium carbonate (1.56 g, 4.77 mmol), and then slowly add methyl iodide (678 mg, 4.77 mmol) dropwise. The reaction mixture was stirred at 25 °C for 18 h. After completion of the reaction, the reaction mixture was quenched by adding water (50 mL), then extracted with ethyl acetate (100 mL × 3). The organic phase was collected and dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate (V / V) = 100:0 to 0:100) to obtain ethyl 3-chloro-1-(1-(ethylsulfonyl)ethyl)-1H-pyrazole-4-carboxylate (630 mg, yield: 90.0%).

[0508] LC-MS, M / Z(ESI): 295.0 [M+H] +

[0509] Step 6: Synthesis of 3-chloro-1-(1-(ethylsulfonyl)ethyl)-1H-pyrazole-4-carboxylic acid

[0510]

[0511] 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), add lithium hydroxide (235 mg, 5.60 mmol). The reaction mixture was stirred at 10 °C for 1 h. After completion of the reaction, the reaction mixture was concentrated to obtain the crude product (260 mg) which was directly used for the next step of the reaction.

[0512] LC-MS, M / Z(ESI): 267.0 [M+H] +

[0513] 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

[0514]

[0515] 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), add (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) were stirred in the reaction solution 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 combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated. The crude product 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 obtain 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%).

[0516] LC-MS, M / Z(ESI): 592.2[M + H] +

[0517] 1 1H NMR (400 MHz, DMSO-d6): δ 8.59 (d, 1H), 8.49 (d, 1H), 8.03 (d, 1H), 7.90 (d, 1H), 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).

[0518] Example 20: Preparation of 5-chloro-N-((1R,3S)-3-((6-chloro-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)-1-((S-methylsulfinimidoyl)methyl)-1H-pyrazole-4-carboxamide (Compound 65)

[0519] The synthetic route of Compound 65 is shown below:

[0520]

[0521] Step 1: Synthesis of Ethyl 5-chloro-1-((S-methylsulfinimidoyl)methyl)-1H-pyrazole-4-carboxylate

[0522]

[0523] At room temperature, dissolve ethyl 5-chloro-1-((methylthio)methyl)-1H-pyrazole-4-carboxylate (500 mg, 2.13 mmol) in anhydrous methanol (20 mL), then successively add iodobenzene diacetate (2.06 g, 6.39 mmol) and ammonium carbonate (614 mg, 6.39 mmol). After the addition is complete, stir the reaction mixture at room temperature for 4 hours. After monitoring by TLC shows that the raw materials have reacted completely, stop stirring, add water (50 mL) for dilution, then extract with ethyl acetate (30 mL×3), collect the organic phase and dry it over anhydrous sodium sulfate. Concentrate the organic phase by distillation under reduced pressure, and purify the residue by silica gel column chromatography (PE:EA = 100:10 (V / V)) to obtain ethyl 5-chloro-1-((S-methylsulfinimidoyl)methyl)-1H-pyrazole-4-carboxylate (500 mg, yield 88.34%).

[0524] LC-MS, M / Z(ESI): 266.0 [M+H] + .

[0525] Step 2: Synthesis of 5-Chloro-1-((S-methylsulfinimido)methyl)-1H-pyrazole-4-carboxylic acid (65-3)

[0526]

[0527] At room temperature, dissolve ethyl 5-chloro-1-((S-methylsulfinimidoyl)methyl)-1H-pyrazole-4-carboxylate (270 mg, 1.02 mmol) in a mixed solution of tetrahydrofuran / methanol / water (1:1:1) (3 mL), then slowly add lithium hydroxide monohydrate (341 mg, 8.13 mmol) under ice bath conditions. Stir the reaction mixture at room temperature for 2 hours. After monitoring by TLC shows that the raw materials have reacted completely, add water (10 mL) to the reaction mixture for dilution, adjust the pH to 4 with dilute hydrochloric acid (1 M), then extract with ethyl acetate (30 mL×3), collect the organic phase and dry it over anhydrous sodium sulfate. Concentrate the organic phase by distillation under reduced pressure to obtain 5-chloro-1-((S-methylsulfinimido)methyl)-1H-pyrazole-4-carboxylic acid (Compound 65-3) (220 mg, yield 91.10%).

[0528] LC-MS, M / Z(ESI): 238.0 [M+H] + .

[0529] Step 3: Synthesis of compound 5-chloro-N-((1R,3S)-3-((6-chloro-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)-1-((S-methylsulfinimidoyl)methyl)-1H-pyrazole-4-carboxamide

[0530]

[0531] Under room temperature conditions, dissolve compound 5-chloro-1-((S-methylsulfinimidoyl)methyl)-1H-pyrazole-4-carboxylic acid (70 mg, 0.29 mmol) in anhydrous N,N-dimethylformamide (1 mL), stir for 10 minutes in an ice bath, then add N,N-diisopropylethylamine (152 mg, 1.18 mmol) and N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)uronium hexafluorophosphate (224 mg, 0.59 mmol), continue to stir in the ice bath for 30 minutes, and then add (1S,3R)-N 1 -(6-chloro-2-(trifluoromethyl)quinolin-4-yl)cyclohexane-1,3-diamine (Intermediate A1) (111 mg, 0.32 mmol). After the addition is complete, slowly warm to room temperature and stir for 6 hours. After monitoring by TLC shows that the raw materials have reacted completely, stop stirring, add water (5 mL) to dilute the reaction solution, then extract with ethyl acetate (5 mL×3), collect the organic phase and dry it with anhydrous sodium sulfate, concentrate the organic phase by distillation under reduced pressure, and purify the residue 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-methylsulfinimidoyl)methyl)-1H-pyrazole-4-carboxamide (65) (50 mg, yield 30.20%).

[0532] LC-MS, M / Z(ESI): 563.0 [M+H] + .

[0533] 1 1H NMR (400 MHz, DMSO-d6): δ 8.60 (d, 1H), 8.16 (d, J = 0.7 Hz, 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).

[0534] 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)

[0535] The synthetic route of Compound 77 is shown as follows:

[0536]

[0537] Step 1: Synthesis of tert-butyl ((1R,3S)-3-((6-fluoro-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)carbamate

[0538]

[0539] At room temperature, 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), then N,N-diisopropylethylamine (621.37 mg, 4.80 mmol) was added, and the reaction mixture was stirred at 100 °C for 12 hours. After the reaction was monitored by TLC and showed completion of the raw materials, stirring was stopped, the reaction mixture was cooled to room temperature, then diluted with water (20 mL), 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 tert-butyl ((1R,3S)-3-((6-fluoro-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)carbamate (295 mg, yield 57.4%).

[0540] LC-MS, M / Z (ESI): 428.6 [M+H] +

[0541] Step 2: Synthesis of (1S,3R)-N 1 -(6-fluoro-2-(trifluoromethyl)quinolin-4-yl)cyclohexane-1,3-diamine hydrochloride

[0542]

[0543] At room temperature, dissolve tert-butyl ((1R,3S)-3-((6-fluoro-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)carbamate (250 mg, 0.58 mmol) in dichloromethane (5 mL). Stir the reaction solution in an ice bath for 10 minutes, then slowly add a 1,4-dioxane solution of hydrogen chloride (1 mol / L, 0.29 mL, 0.29 mmol). The reaction solution is slowly warmed to room temperature and stirred for 2 hours. After monitoring by TLC shows that the raw material reaction is complete, stop stirring. Concentrate the reaction solution 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), which is directly used in the next step of the reaction.

[0544] LC-MS, M / Z(ESI): 328.5[M+H] +

[0545] 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

[0546]

[0547] At room temperature, dissolve 3-chloro-1-(1-(methylsulfonyl)ethyl)-1H-pyrazole-4-carboxylic acid (60 mg, 0.24 mmol) in anhydrous N,N-dimethylformamide (2 mL), then add N,N-diisopropylethylamine (122.76 mg, 0.95 mmol) and N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)uronium hexafluorophosphate (180.6 mg, 0.47 mmol). Stir at room temperature for 30 minutes, then add (1S,3R)-N 1 -(6-fluoro-2-(trifluoromethyl)quinolin-4-yl)cyclohexane-1,3-diamine hydrochloride (93.3 mg, 0.28 mmol). Continue to stir the reaction solution at room temperature for 4 hours. After monitoring by TLC shows that the raw material reaction is complete, stop stirring. Cool the reaction solution to room temperature, then add water (10 mL) for dilution, extract with ethyl acetate (5 mL×3). Combine the organic phases, dry with anhydrous sodium sulfate, filter and concentrate. The residue is separated and purified by silica gel column chromatography (petroleum ether: ethyl acetate (V / V)=100:30) to obtain 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%).

[0548] LC-MS, M / Z (ESI): 562.1 [M+H] +

[0549] 1 H NMR (400 MHz, DMSO-d6) δ 8.47 (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).

[0550] 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

[0551] 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 (250 mg, 0.445 mmol) was purified by SFC (Daicel ChiralPak IC, 40 mm I.D. × 250 mm, 10 μm, mobile phase: A = n-hexane, B = ethanol; gradient: 30%, 1.68 h) to obtain 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).

[0552] Compound 77-A: LC-MS, M / Z (ESI): 562.2 [M+H] + ;

[0553] 11H NMR (400 MHz, DMSO-d6): δ 8.47 (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).

[0554] Compound 77-B: LC-MS, M / Z (ESI): 562.2 [M + H] +

[0555] 1 1H NMR (400 MHz, DMSO-d6): δ 8.47 (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).

[0556] 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)

[0557] The synthetic route of Compound 80 is as follows:

[0558]

[0559] The first step: Synthesis of tert-butyl ((1R,3S)-3-((6-methyl-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)carbamate

[0560]

[0561] At room temperature, 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). N,N-Diisopropylethylamine (1.05 g, 8.14 mmol) was added. After the addition was completed, the reaction mixture was stirred at 100 °C for 8 hours. After monitoring by TLC showed that the raw materials were completely reacted, the stirring was stopped. The reaction mixture was cooled to room temperature, then diluted with water (30 mL) and extracted with ethyl acetate (10 mL×3). The organic phase was collected, 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)=80:20) to obtain tert-butyl ((1R,3S)-3-((6-methyl-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)carbamate (400 mg, yield 46.4%).

[0562] LC-MS,M / Z(ESI):424.6(M+H + )

[0563] Step 2: Synthesis of (1S,3R)-N 1 -(6-methyl-2-(trifluoromethyl)quinolin-4-yl)cyclohexane-1,3-diamine hydrochloride

[0564]

[0565] At room temperature, tert-butyl ((1R,3S)-3-((6-methyl-2-(trifluoromethyl)quinolin-4-yl)amino)cyclohexyl)carbamate (400 mg, 0.94 mmol) was dissolved in dichloromethane (5 mL). The mixture was stirred at 0 °C for 10 minutes, and then a 1,4-dioxane solution of hydrogen chloride (1 M, 0.47 mL, 0.47 mmol) was slowly added dropwise. After the addition was completed, the reaction mixture was allowed to warm to room temperature and stirred for 2 hours. After monitoring by TLC showed that the raw materials were completely reacted, the stirring was stopped. The solvent was removed by evaporation under reduced pressure to obtain (1S,3R)-N 1 -(6-methyl-2-(trifluoromethyl)quinolin-4-yl)cyclohexane-1,3-diamine hydrochloride (350.00 mg, crude product).

[0566] LC-MS,M / Z(ESI):323.5(M+H + )

[0567] 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

[0568]

[0569] At room temperature, dissolve 3-chloro-1-(1-(methylsulfonyl)ethyl)-1H-pyrazole-4-carboxylic acid (80 mg, 0.32 mmol) in anhydrous N,N-dimethylformamide (2 mL), then add N,N-diisopropylethylamine (163.68 mg, 1.27 mmol) and N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)uronium hexafluorophosphate (240.78 mg, 0.63 mmol). Stir the reaction mixture at room temperature for 30 minutes, then add (1S,3R)-N 1 -(6-methyl-2-(trifluoromethyl)quinolin-4-yl)cyclohexane-1,3-diamine hydrochloride (170.89 mg, 0.47 mmol), and continue to stir at room temperature for 4 hours. After monitoring by TLC shows that the raw materials have reacted completely, stop stirring, add water (10 mL) for dilution, extract with ethyl acetate (5 mL×3), collect the organic phase and dry it over anhydrous sodium sulfate, filter and concentrate. The residue is separated and purified by silica gel column chromatography (petroleum ether: ethyl acetate (V / V) = 100:40) to obtain 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%).

[0570] LC-MS, M / Z (ESI): 558.1 (M+H + )

[0571] 1 H NMR (400 MHz, CDCl3) δ 8.23 (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).

[0572] 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)

[0573] The synthetic route of compound 86 is as follows:

[0574]

[0575] At room temperature, dissolve compound 3-chloro-1-((methylsulfonyl)methyl)-1H-pyrazole-4-carboxylic acid (65.0 mg, 0.27 mmol) in anhydrous N,N-dimethylformamide (2 mL), then add N,N-diisopropylethylamine (140.81 mg, 1.09 mmol) and N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)uronium hexafluorophosphate (178.81 mg, 0.54 mmol). Stir the reaction mixture at room temperature for 30 minutes, then add (1S,3R)-N 1 -(6-Fluoro-2-(trifluoromethyl)quinolin-4-yl)cyclohexane-1,3-diamine hydrochloride (118.90 mg, 0.32 mmol), and continue to stir at room temperature for 4 hours. After monitoring by TLC shows that the raw materials have reacted completely, stop stirring. Dilute the reaction mixture with water (10 mL), extract with ethyl acetate (5 mL × 3), collect the organic phase and dry it over anhydrous sodium sulfate. Concentrate the organic phase under reduced pressure by distillation. Purify the residue by silica gel column chromatography (petroleum ether: ethyl acetate (V / V) = 100:30) to obtain 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, yield 42.5%).

[0576] LC-MS, M / Z (ESI): 548.0 (M + H + )

[0577] 1 1H NMR (400 MHz, DMSO-d6) δ 8.36 (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).

[0578] The preparation method of the following compounds refers to Preparation Example 1,

[0579]

[0580]

[0581]

[0582]

[0583]

[0584]

[0585]

[0586]

[0587]

[0588]

[0589]

[0590]

[0591]

[0592]

[0593]

[0594]

[0595]

[0596]

[0597] Test Example 1: In vitro Calcium Flux Assay of MRGPRX2

[0598] Determination of the antagonistic effect of the compound on MRGPRX2 was carried out in a CHO stable cell line highly expressing human MRGPRX2. 18 hours before the experiment, cells were seeded at a certain density in a black-walled transparent bottom plate containing DMEM / F12 (1:1) medium and incubated at 37 °C in 5% CO2 for 18 hours. Then, the corresponding amount of dye solution was added to each well of the cells, and the plate was returned to the 37 °C incubator for continued incubation in the dark for 30 min. Then, it was incubated in the dark at room temperature for 10 min. Subsequently, different final concentrations of the compound were added to each well and allowed to equilibrate for 20 min. Finally, a certain amount of C48 / 80 solution (sigma, lot: 0000197124) was added to the cells, and the fluorescence signal value was detected using FLIPR. With the compound concentration on the X-axis and the fluorescence signal value on the Y-axis, the antagonistic effect (IC 50 value) of the compound was calculated using the software GraphPad Prism 8.0. The experimental results are shown in Table 1.

[0599] Table 1: Results of the MRGPRX2 calcium influx assay

[0600]

[0601]

[0602] The results of the MRGPRX2 calcium influx assay indicate that the compound of the present invention has a good antagonistic effect on MRGPRX2.

[0603] Test Example 2: Test for the cytotoxicity of the compound to hepatocytes

[0604] The cytotoxicity test of the compound on hepatocytes was carried out on HepG2 (ATCC, HB - 8065) cells. The cell viability was measured using the CellTiter - Glo Luminescent Cell Viability Assay kit (Promega, G7573). The toxicity of the compound was characterized by the inhibition of the viability of HepG2 cells. Logarithmic - phase HepG2 cells were collected, the concentration of the cell suspension was adjusted, and plated in a 96 - well cell culture plate at 5000 cells / well. The cells were incubated overnight in a cell culture incubator with 5% CO2 at 37 °C. The next day, the culture medium was changed and different concentrations of the compound solution were added. At the same time, a negative control group (cells + DMSO) and a blank control group (culture medium + DMSO) were set up, and incubated in a cell culture incubator with 5% CO2 at 37 °C for 72 hours. After the treatment, the operation was carried out according to the kit instructions, and the luminescence signal values in different wells were detected on an EnVision plate reader (2104). The viability inhibition of the compound at different concentrations on HepG2 cells was calculated according to the following formula. With the compound concentration as the X - axis and the inhibition rate as the Y - axis, the toxic effect of the compound on HepG2 (IC 50 value) was calculated by the software GraphPad Prism 8.0.

[0605]

[0606] The results of the hepatocyte cytotoxicity test showed that the compound of the present invention exhibited good safety and low hepatocyte toxicity.

[0607] Test Example 3: Pharmacokinetics Test in Mice

[0608] For the pharmacokinetics test in mice, male ICR mice, weighing 20 - 25 g, were fasted overnight. Three mice were taken and administered orally by gavage at a dose of 10 mg / kg. Blood samples were collected before dosing and at 15, 30 minutes, and 1, 2, 4, 8, 24 hours after dosing. The blood samples were centrifuged at 6800 g at 2 - 8 °C for 6 minutes, the plasma was collected and stored at - 80 °C. Plasma samples at each time point were taken, mixed with 3 - 5 volumes of acetonitrile solution containing an internal standard, vortex - mixed for 1 minute, centrifuged at 13000 rpm at 4 °C for 10 minutes, the supernatant was taken and mixed with 3 volumes of water, and an appropriate amount of the mixed solution was subjected to LC - MS / MS analysis. The main pharmacokinetic parameters were analyzed using the non - compartmental model of WinNonlin 7.0 software.

[0609] Table 2: Results of the Pharmacokinetics Test in Mice

[0610]

[0611] The results of the mouse pharmacokinetic experiment showed that the compound of the present invention exhibited excellent pharmacokinetic properties and good drug-likeness.

[0612] Test Example 4: Rat Pharmacokinetic Experiment

[0613] For the rat pharmacokinetic experiment, male SD rats weighing 180 - 240 g were used and fasted overnight. Three rats were selected and administered 10 mg / kg by oral gavage. Blood samples were collected before dosing and at 15, 30 minutes, and 1, 2, 4, 8, 24 hours after dosing. The blood samples were centrifuged at 6800 g for 6 minutes at 2 - 8 °C, the plasma was collected and stored at -80 °C. Plasma samples at each time point were mixed with 3 - 5 volumes of acetonitrile solution containing internal standard, vortexed for 1 minute, centrifuged at 13000 rpm for 10 minutes at 4 °C, the supernatant was mixed with 3 volumes of water, and an appropriate amount of the mixture was subjected to LC-MS / MS analysis. The main pharmacokinetic parameters were analyzed using the non-compartmental model of WinNonlin 7.0 software.

[0614] The results of the rat pharmacokinetic experiment showed that the compound of the present invention exhibited excellent rat pharmacokinetic properties and good drug-likeness.

[0615] Test Example 5: Dog Pharmacokinetic Experiment

[0616] For the dog pharmacokinetic experiment, male Beagle dogs weighing 8 - 10 kg were used and fasted overnight. Three Beagle dogs were selected and administered 5 mg / kg by oral gavage. Blood samples were collected before dosing and at 15, 30 minutes, and 1, 2, 4, 8, 24 hours after dosing. The blood samples were centrifuged at 6800 g for 6 minutes at 2 - 8 °C, the plasma was collected and stored at -80 °C. Plasma samples at each time point were mixed with 3 - 5 volumes of acetonitrile solution containing internal standard, vortexed for 1 minute, centrifuged at 13000 rpm for 10 minutes at 4 °C, the supernatant was mixed with 3 volumes of water, and an appropriate amount of the mixture was subjected to LC-MS / MS analysis. The main pharmacokinetic parameters were analyzed using the non-compartmental model of WinNonlin 7.0 software.

[0617] Table 5 Results of Dog Pharmacokinetic Experiment

[0618]

[0619]

[0620] The results of the dog pharmacokinetic experiment showed that the compound of the present invention exhibited excellent dog pharmacokinetic properties and good drug-likeness.

[0621] Test Example 6: Determination of the Inhibitory Effect of the Compound on the BSEP Bile Efflux Transporter

[0622] The inhibitory effect of the compound on the bile salt export pump (BSEP) was tested using vesicles (GenoMembrane) expressing the human BSEP bile efflux transporter. Different concentrations of the compound were pre-incubated with the vesicles for 5 minutes. A negative control (NC) group and a positive control (PC) group were set up: in the NC group, the vesicles were pre-incubated with blank buffer at 37 °C for 5 minutes, and in the PC group, the positive inhibitor was pre-incubated with the vesicles at 37 °C for 5 minutes. Subsequently, under the conditions of adding ATP or AMP respectively, they were incubated with the probe substrate at 37 °C for 5 minutes. The experiment was terminated with pre-cooled Buffer B1 (10×Buffer B1 (Stopping and Washing Buffer): 100 mM Hepes-Tris, 1000 mM KNO3, 500 mM Sucrose). The test samples were transferred to a 96-well filter plate, filtered by a vacuum pump, then washed repeatedly 5 times with 0.2 mL of pre-cooled Buffer B1, the vesicles on the filter plate were dissolved with 50 μL of 80% methanol, and after collection, the filtrate was collected by centrifugation at 2000 rpm for 2 minutes. This was repeated once, and the filtrates of the two times were combined and mixed to obtain approximately 100 μL of filtrate. Pre-cooled methanol containing the internal standard was added, and centrifuged at 12,000 rpm for 5 min. The supernatant was taken for LC-MS / MS quantitative detection of the content of the transported substrate. With the compound concentration on the X-axis and the relative activity (% of NC) on the Y-axis, the IC 50 value and the inhibition rate of the compound inhibiting the bile efflux transporter activity were calculated by the software Prism.

[0623] The transport rate (activity) and relative activity under different conditions were calculated according to the following formula:

[0624]

[0625] The results of the inhibitory test on the BSEP bile efflux transporter 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.

[0626] Test Example 7: Thermodynamic Solubility Test

[0627] Phosphate buffer (PBS) with pH 7.4 was prepared. The compound was accurately weighed and added to the prepared phosphate buffer with pH 7.4 to prepare a solution with a concentration of 4 mg / mL, shaken at a speed of 1000 rpm for 1 hour, and then incubated overnight at room temperature. The incubated solution was centrifuged at 12000 rpm for 10 minutes to remove undissolved particles, and the supernatant was transferred to a new centrifuge tube. After appropriate dilution of the supernatant, acetonitrile solution containing the internal standard was added, and quantification was carried out using a standard curve prepared with the same matrix.

[0628] The results of the thermodynamic solubility test show that the compounds of the present invention have good thermodynamic solubility and good drug-likeness.

[0629] Test Example 8: Human Liver Microsome Stability Test

[0630] The human liver microsome stability test was performed by co-incubating the compound with human liver microsomes in vitro. First, the test compound was prepared as a 10 mM stock solution in DMSO, and then the compound was diluted to 0.5 mM with acetonitrile. Human liver microsomes (Corning) were diluted with PBS to form a microsome / buffer solution, and this solution was used to dilute the 0.5 mM compound to form a working solution with a compound concentration of 1.5 μM and a human liver microsome concentration of 0.75 mg / ml in the working solution. Take a deep well plate, add 30 μL of the working solution to each well, then add 15 μL of pre-warmed 6 mM NADPH solution to initiate the reaction, and incubate at 37 °C. At 0, 5, 15, 30, and 45 minutes of incubation, 135 μL of acetonitrile was added to the corresponding wells to terminate the reaction. After terminating the reaction with acetonitrile at the last 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 taken, purified water was added in a 1:1 ratio, and then LC-MS / MS detection was performed to obtain the ratio of the compound peak area to the internal standard peak area at each time point. The ratio of the compound peak area at 5, 15, 30, and 45 minutes to the peak area at 0 minutes was compared, and the remaining percentage of the compound at each time point was calculated. Graphpad 5 software was used to calculate T 1 / 2 。

[0631] The results of the human liver microsome stability test show that the compounds of the present invention exhibit excellent human liver microsome stability and good drug-likeness.

[0632] Test Example 9: Inhibition Test of Compounds on Cytochrome P450

[0633] The inhibitory potential of the compound on cytochrome P450 (CYP450) subtype CYP3A4 (two substrates, midazolam and testosterone) was detected. First, the test compound was prepared as a 10 mM stock solution in DMSO, and the CYP3A4 inhibitor ketoconazole was prepared as 10 mM, 2.5 mM, and 2.5 mM stock solutions in DMSO. The test compound and ketoconazole were diluted with acetonitrile to a final concentration of 400-fold (compound: 10 μM, ketoconazole: 2.5 μM).

[0634] A 4-fold final concentration of NADPH cofactor (66.7 mg NADPH was added to 10 mL of potassium phosphate buffer) and substrates were prepared with potassium phosphate buffer (0.1 M, pH 7.4). The final concentration of the CYP3A4 substrate midazolam was 320 μM, and the final concentration of the CYP3A4 substrate testosterone was 20 μM.

[0635] Prepare a human liver microsome solution with potassium phosphate buffer on ice at a concentration of 0.2 mg / mL. Prepare solutions of the test compound and control inhibitor at twice the final concentration using the human liver microsome solution on ice. Add 30 μL of the test compound and control inhibitor solutions to the test wells respectively, and add 15 μL of the substrate, and perform duplicate well operations. Incubate a 96-well assay plate and NADPH solution at 37 °C for 5 minutes, and add 15 μL of pre-warmed 8 mM NADPH solution to the assay plate to initiate the reaction. The CYP3A4 assay plate is pre-incubated at 37 °C for 5 minutes. Add 120 μL of acetonitrile to terminate the reaction. 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, take the supernatant, add purified water at a ratio of 1:1 and then perform LC-MS / MS detection to obtain the ratio of the compound peak area to the internal standard peak area. Compare the peak area ratio of the compound with the peak area ratio of the control inhibitor, and calculate the inhibition rate.

[0636] Table 6 Inhibition data of 10 μM compound on cytochrome P450 (CYP450) subtype CYP3A4 (two substrates midazolam and testosterone)

[0637] Inhibition rate of CYP3A4 (midazolam) (%) Inhibition rate of CYP3A4 (testosterone) (%) Compound 77 26.3 31.7 Compound 46 16.5 28.4

[0638] 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 drug-likeness.

[0639] Test Example 10: Evans blue vascular permeability test

[0640] For the Evans blue vascular permeability test, C57 mice are used. The left hind paw of the mouse is used as the model group, and the right hind paw is used as the negative control group. Different doses of the drug are administered by gavage. 2 h after administration, inject a 0.4% Evans blue (Sigma-Aldrich, Lot#SHBP1253) solution into the tail vein. 5 - 10 min later, inject a C48 / 80 (Sigma-Aldrich, Lot#C2313) solution subcutaneously into the left hind paw, and inject normal saline into the right hind paw. The mice are sacrificed 15 min later. The thickness of the left and right hind paws of the mice is measured with a vernier caliper. Cut off the mouse's feet, dry them, and weigh them. Crush the paws, dissolve the Evans blue in the paws with acetone: normal saline (7:3), and measure the absorbance at 620 nm to calculate the absorbance per unit volume. Calculate the ear swelling rate and the amount of Evans blue exudation per unit mass.

[0641] The results of the Evans blue vascular permeability test show that the compound of the present invention exhibits good drug efficacy, can significantly inhibit C48 / 80-induced vascular leakage, and has good drug-likeness.

[0642] The above has given an exemplary description of the implementation modes of the technical solution of the present disclosure. It should be understood that the protection scope of the present disclosure is not limited to the above implementation modes. Any modifications, equivalent replacements, improvements, etc. made by those skilled in the art within the spirit and principle of the present disclosure shall be included within the protection scope of the claims of this application.

Claims

1. A compound represented by formula I-A, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug: 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 In, 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. When the substituents R c are multiple, the substituents are the same or different; Ring B is -(CH2) 0-2 -C 3-12 cycloalkyl, -(CH2) 0-2 -3- to 10-membered hetero cycloalkyl or -(CH2) 0-2 -5- to 10-membered heteroaryl; Ring C is a 6- to 10-membered aryl or 5- to 10-membered heteroaryl; R a and R c each independently is H, 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 haloalkyl, C 3-8 halocycloalkyl, C 1-6 alkylhydroxyl, C 1-6 alkylcarbonyl, C 1-6 alkoxy, C 1-6 haloalkoxy or a 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 alkylhydroxyl, 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, 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; 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 with one or more R d ; said 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, 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 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 , 6-10 membered aryl, C 3-8 cycloalkyl, 5-10 membered heteroaryl or 5-10 membered heterocycloalkyl, or two R f groups together with the atoms to which they are attached form a 5-11 membered heterocycloalkyl; said R g is H, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 3-8 cycloalkyl, 6-10 membered aryl, 5-10 membered heteroaryl or 5-10 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 hydroxyl, a 5- to 10-membered heterocycloalkyl alone, or when there are two of said R k 's, they form a 3- to 10-membered heterocycloalkyl with the N, S or P atom to which they are attached; said amino, 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 hydroxyl, 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 's; 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 's, said R m 's are the same or different; m and r are each 0, 1, 2 or 3.

2. The compound of formula I-A as claimed in claim 1, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug, characterized in that, It has the structure shown in formula II-A, Among them, 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. When there are multiple substituents R c , the substituents are the same or different; Ring C is a 6- to 10-membered aryl or 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 a 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 a 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 halocycloalkyl, C 1-6 alkylhydroxyl, 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 1-6 haloalkyl, C 3-8 halocycloalkyl, C 1-6 alkylhydroxyl, 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 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, 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 hydroxy, a 5- to 10-membered heterocycloalkyl alone, or when 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; said amino, 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, 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 ; 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 , 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 as described in claim 1, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug, characterized in that, Selected from and / or is 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 as claimed in 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 as claimed in claim 1, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug, characterized in that, The said 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 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, is 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 definition described in any one of claims 1 to 6.

8. The compound represented by Formula I-A as described in claim 1, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug, characterized in that, The compound includes: 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 compound includes: And / or, the compound is selected from any one of the following compounds:

10. A pharmaceutical composition, characterized in that, The pharmaceutical composition includes: 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 described in claim 10, the use includes: Antagonizing MRGPRX2; And / or, preventing and / or treating MRGPRX2-related diseases; And / or, preparing a drug, pharmaceutical composition or preparation for antagonizing MRGPRX2, and / or preventing and / or treating MRGPRX2-related diseases.

12. The use according to claim 11, wherein The MRGPRX2-related diseases 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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