Sulfonamide derivative and use thereof

Abstract

The present invention relates to a sulfonamide derivative and a use thereof, and in particular to a compound of general formula (I) or general formula (I-1) or a stereoisomer, a tautomer, a racemate, a pharmaceutically acceptable salt thereof, an intermediate thereof and a preparation method therefor, and a use thereof in the preparation of a drug for treating Nav1.8-related diseases.

Description

A sulfonamide derivative and its application Technical Field

[0001] This invention relates to a sulfonamide compound of general formula (I) or its stereoisomers, tautomers, racemates, pharmaceutically acceptable salts, intermediates thereof, and methods of preparation thereof, as well as its use in the preparation of medicaments for treating or relieving pain. Background Technology

[0002] Pain originates from nociceptors in the peripheral nervous system. These receptors convert perceived thermal, mechanical, or chemical stimuli into nerve impulses (action potentials), which are then transmitted via afferent nerve fibers to the cell body of the dorsal root ganglion (DRG), ultimately reaching higher nerve centers and causing pain sensation. The generation and conduction of action potentials in neurons depend on voltage-gated sodium channels (VGSCS) on the cell membrane. When the cell membrane depolarizes, sodium channels are activated, opening and causing an influx of sodium ions, further depolarizing the cell membrane and leading to the generation of action potentials.

[0003] VGSCS consists of a porous α-subunit (approximately 260 kDa) and an associated smaller β-subunit (30–40 kDa). The associated α-subunit family comprises 10 members, nine of which (Nav1.1–1.9) are voltage-gated. Nav1.8 is encoded by the gene SCN10A and is preferentially expressed in peripheral sensory neurons. It has been shown to shape action potentials in these neurons. Nav1.8 transcripts and proteins have been found in dorsal root ganglion (DRG) neurons. Nav1.8 has not been detected in non-neuronal tissues (e.g., heart and skeletal muscle) or the central nervous system (including the brain and spinal cord).

[0004] The crucial role of Nav1.8 in pain signal transduction has been supported by multiple pieces of evidence. Based on a series of animal studies and human genetic evidence, selective inhibition of Nav1.8 has the potential to become a novel analgesic therapy. Summary of the Invention

[0005] The purpose of this invention is to provide a class of sulfonamide derivatives or pharmaceutically acceptable salts thereof for use as Nav1.8 inhibitors. The compounds of this invention effectively inhibit Nav1.8 and can be used to treat diseases such as pain. They exhibit good selectivity for Nav1.8 within the sodium ion channel family, show low activity in various non-sodium ion channel family receptors or enzymes (agonists or inhibitors), demonstrate good analgesic activity in animals, show good safety in in vivo and in vitro screening, have no significant CYP3A4 induction, exhibit weak inhibition of UGT1A1, and possess good solubility and stability.

[0006] This invention provides a compound of general formula (I) or general formula (I-1) or its stereoisomers, deuterated derivatives, solvates, prodrugs, metabolites, pharmaceutically acceptable salts, or cocrystals.

[0007] In some embodiments, the compound represented by formula (I) is selected from formula (Ia):

[0008] In some embodiments, the compound represented by general formula (I) is selected from general formula (Ia-1):

[0009] In some embodiments, the compound represented by general formula (I) is selected from general formula (Ia-2):

[0010] In some embodiments, the compound represented by formula (I) is selected from formula (Ib).

[0011] In some implementation schemes, R b Selected from R B ;

[0012] In some implementations, X is selected from O, S, -S(O)- or -S(O)2;

[0013] In some implementation schemes, X is selected from O and S;

[0014] In some implementations, Q is selected from...

[0015] In some implementations, Q1 is selected from C. 6-10 Aryl, 5- to 10-membered heteroaryl, C 5-10 Carbon rings, 5- to 10-membered heterocycles or The aryl, heteroaryl, carbocyclic, or heterocyclic group is optionally surrounded by 1 to 5 R groups. q replace;

[0016] In some implementations, Q1 is selected from phenyl, benzo[a]C4-6 Carbocyclic, benzo[4-6] heterocyclic, 5-6 fused heteroaryl, 5-6 fused heterocyclic, 8-10 fused heteroaryl or Q1 is arbitrarily selected by 1 to 4 Rs q replace;

[0017] In some implementations, Q1 is selected from Q1 is arbitrarily selected by 1 to 4 Rs q replace;

[0018] In some implementations, Q1 is selected from Q1 is arbitrarily selected by 1 to 3 Rs q replace;

[0019] In some implementations, R is selected from C. 1-6 alkyl, -C(=O)R q1 -P(=O)R q1 R q2 -C 1-4 Alkylene-OP(=O)R q1 R q2 -C(O)OC 1-4 Alkylene-OP(=O)R q1 R q2 -S(=O)2R q1 C 3-6 Carbocyclic, 4- to 7-membered heterocyclic, wherein the alkyl, alkylene, carbocyclic or heterocyclic is optionally surrounded by 1 to 4 R... k replace;

[0020] In some implementations, R is selected from C. 1-4 alkyl, -C(=O)R q1 -P(=O)R q1 R q2 -C 1-2 Alkylene-OP(=O)R q1 R q2 -C(O)OC 1-2 Alkylene-OP(=O)R q1 R q2 -S(=O)2R q1 C 3-6 Carbocyclic, 4- to 7-membered heterocyclic, wherein the alkyl, alkylene, carbocyclic or heterocyclic is optionally surrounded by 1 to 4 R... k replace;

[0021] In some implementations, R is selected from -C(=O)R q1 -P(=O)R q1 R q2 -CH2-OP(=O)R q1 R q2 -C(O)O-CH2-OP(=O)R q1 R q2 -S(=O)2R q1 C 3-6 Carbon rings, 4- to 7-membered heterocycles, wherein the CH2, carbon rings, or heterocycles are optionally surrounded by 1 to 4 R... k replace;

[0022] In some implementations, R is selected from C(=O)R q1 -P(=O)R q1 R q2 -CH2-OP(=O)R q1 R q2 -C(O)O-CH2-OP(=O)R q1 R q2 -S(=O)2R q1 The CH2 is optionally converted by 1 to 2 R k replace;

[0023] In some implementations, R is selected from

[0024] In some implementation schemes, R q1 R q2 Each is independently selected from H, OH, and C. 1-6 Alkyl, C 1-6 Alkoxy, C 3-6 Carbon ring, -OC 3- A 6-membered carbon ring, a 4- to 7-membered heterocycle, wherein the alkyl, alkoxy, carbocyclic or heterocyclic ring is optionally surrounded by 1 to 4 R... k replace;

[0025] In some implementation schemes, R q1 R q2 Each is independently selected from H, OH, and C. 1-4 Alkyl, C 1-4 Alkoxy, C 3-6 Carbon ring, -OC 3- A 6-membered carbon ring, a 4- to 7-membered heterocycle, wherein the alkyl, alkoxy, carbocyclic or heterocyclic ring is optionally surrounded by 1 to 4 R... k replace;

[0026] In some implementation schemes, R q1 Rq2 Each is independently selected from H, OH, or arbitrarily selected by 1 to 4 R. k The substitution is made with one of the following groups: methyl, ethyl, propyl, butyl, isopropyl, tert-butyl, methoxy, ethoxy, isopropyloxy, tert-butyloxy, cyclopropyl, cyclobutyl, cyclopentyl, oxecyclobutyl, oxecyclopentyl, oxecyclohexyl, azircyclobutyl, azircyclopentyl, azircyclohexyl;

[0027] In some implementation schemes, R q1 R q2 Direct connection forms a 4- to 7-membered heterocycle, wherein the heterocycle is optionally bounded by 1 to 4 R... k replace;

[0028] In some implementation schemes, R q3 R q4 Each is independently selected from H or arbitrarily selected by 1 to 4 Rs. k One of the following groups is substituted: C 1- 6-alkyl, -C 1-6 Alkylene -NH2, -C 1-6 Alkylene -C(=O)NH2, -C 1-6 Alkylene -NHC(=NH)NH2, -C 1-6 Alkylene -NHC(=O)NH2、-C 1-6 alkylene -OH, -C 1-6 Alkylene -SH, -C 1-6 Alkylene -CO2H, -C 1-6 Alkylene-C 3-10 Carbon ring, C 3-10 Carbon ring, -C 1-6 Alkylene-3 to 10-membered heterocycles, 3 to 10-membered heterocycles;

[0029] In some implementation schemes, R q3 R q4 Each is independently selected from H or arbitrarily selected by 1 to 4 Rs. k One of the following groups is substituted: C 1- 4-alkyl, -C 1-4 Alkylene -NH2, -C 1-4 Alkylene -C(=O)NH2, -C 1-4 Alkylene -NHC(=NH)NH2, -C 1-4 Alkylene -NHC(=O)NH2、-C 1-4 alkylene -OH, -C 1-4 Alkylene -SH, -C 1-4 Alkylene -CO2H, -C 1-4 Alkylene-C 6-10 Aryl, C 6-10 Aryl, -C1-4 alkylene-5 to 10-membered heteroaryl, 5 to 10-membered heteroaryl, C 3-6 cycloalkyl, 4- to 7-membered heterocyclic groups, -CH2-C 3-6 Cycloalkyl groups, -CH2-4 to 7-membered heterocyclic groups;

[0030] In some implementation schemes, R q3 R q4 Each is independently selected from H or arbitrarily selected by 1 to 4 Rs. k The substitution is performed using one of the following groups: methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, -CH2NH2, -CH2CH2NH2, -CH2CH2CH2NH2, -CH2CH2CH2CH2NH2, -CH2NHC(=NH)NH2, -CH2CH2NHC(=NH)NH2, -CH2CH2CH2NHC(=NH)NH2, -CH2CH2CH2NHC(=O)NH2, -CH2OH, -CH2SH, -CH2CH2SH, -CH2CO2H, -CH2 CH2CO2H, -CH2C(=O)NH2, -CH2CH2C(=O)NH2, -CH2CH2CH2C(=O)NH2, -CH2CH2CH2CH2C(=O)NH2, -CH2-phenyl, -CH2-indole, -CH2-5 to 10-membered heteroaryl, -CH2-C 3-6 cycloalkyl, C 3- 6-membered cycloalkyl, 4- to 7-membered heterocyclic alkyl, or -CH2-4- to 7-membered heterocyclic alkyl;

[0031] In some implementation schemes, R q3 R q4 Each is independently selected from H or arbitrarily selected by 1 to 4 Rs. kThe substitution is made with one of the following groups: methyl, ethyl, propyl, butyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, -CH2NH2, -CH2CH2NH2, -CH2CH2CH2NH2, -CH2CH2CH2CH2NH2, -CH2NHC(=NH)NH2, -CH2CH2NHC(=NH)NH2, -CH2CH2CH2NHC(=NH)NH2, -CH2CH2CH2NHC(=O)NH2, -CH2OH, -CH2SH, -CH2CH2SH, -CH2CO2H, -CH2 CH2CO2H, -CH2-phenyl, -CH2-imidazolyl, -CH2-indolyl, cyclopropyl, cyclobutyl, cyclopentyl, oxecyclobutyl, oxecyclopentyl, oxecyclohexyl, azircyclobutyl, azircyclopentyl, azircyclohexyl, -CH2-cyclopropyl, -CH2-cyclobutyl, -CH2-cyclopentyl, -CH2-oxecyclobutyl, -CH2-oxecyclopentyl, -CH2-azircyclobutyl, -CH2-azircyclopentyl, -CH2-azircyclohexyl;

[0032] In some implementation schemes, R q3 R q4 Direct connection forms C 3-6 Carbon rings or 4- to 7-membered heterocycles, wherein the carbon rings or heterocycles are optionally surrounded by 1 to 4 R... k replace;

[0033] In some implementation schemes, R q3 R q4 Direct connection forms C 3-6 Cycloalkyl or 4- to 7-membered heterocycloalkyl, wherein the cycloalkyl or heterocycloalkyl is optionally surrounded by 1 to 4 R... k replace;

[0034] In some implementation schemes, R q3 R q4 Directly linked to form cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, oxacyclobutyl, oxacyclopentyl, oxacyclohexyl, aziridine, aziridinepentyl, and aziridinehexyl, wherein the cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, oxacyclobutyl, oxacyclopentyl, oxacyclohexyl, aziridine, aziridinepentyl, and aziridinehexyl are optionally separated by 1 to 4 R... k replace;

[0035] In some implementation schemes, B is selected from C. 6-10 Aryl, 5- to 10-membered heteroaryl, C 5-10 Carbocyclic rings, 5- to 10-membered heterocyclic rings, wherein the aryl, heteroaryl, carbocyclic, or heterocyclic rings are optionally surrounded by 1 to 5 R groups. B replace;

[0036] In some implementations, B is selected from phenyl, benzo[C] 4-6 Carbocyclic, benzo[4- to 6-membered heterocyclic, 5- to 6-membered heteroaryl, 8- to 10-membered fused-ring heteroaryl, wherein B is optionally surrounded by 1 to 4 R[]. B replace;

[0037] In some implementation schemes, B is selected from Or phenyl, wherein the B is optionally surrounded by 1 to 4 R B replace;

[0038] In some implementation schemes, B is selected from

[0039] In some implementation schemes, B is selected from

[0040] In some implementation schemes, R 1 R 2 R 3 R 4 R 5 Each is independently selected from H, halogens, CN, OH, and C. 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, -OC 1-6 Alkyl, -SC 1-6 Alkyl, C 3-6 Carbocyclic, 3- to 7-membered heterocyclic, wherein the alkyl, alkenyl, alkynyl, carbocyclic or heterocyclic is optionally surrounded by 1 to 4 R... k replace;

[0041] In some implementation schemes, R 1 R 2 R 3 R 4 R 5 Each of the following groups is independently selected from H, F, Cl, Br, cyano, methyl, ethyl, methoxy, ethoxy, methylthio, cyclopropyl, cyclobutyl, vinyl, and ethynyl, wherein the methyl, ethyl, methoxy, ethoxy, methylthio, cyclopropyl, cyclobutyl, vinyl, and ethynyl groups are optionally prefixed with 1 to 4 R groups. k replace;

[0042] In some implementation schemes, R 1 R 2 R 3 R 4 R 5 Each is independently selected from H, methyl, ethyl, CH2F, CHF2, CF3;

[0043] In some implementation schemes, R 1 R 4 Direct connection forms C 3-6 Carbon rings or 4- to 7-membered heterocycles, wherein the carbon rings or heterocycles are optionally surrounded by 1 to 6 R... k replace;

[0044] In some implementation schemes, R 1 R 4 Direct connection forms C 3-6 Carbon rings or 4- to 7-membered heterocycles, wherein the carbon rings or heterocycles are optionally surrounded by 1 to 4 R... k replace;

[0045] In some implementation schemes, R 1 R 4 Direct connection forms C 3-6 Carbon ring, wherein the carbon ring is optionally divided by 1 to 4 R k replace;

[0046] In some implementation schemes, R q R B Each is independently selected from H, deuterium, =O, halogen, CN, OH, NH2, NHC 1-6 Alkyl, N(C) 1-6 Alkyl)2, C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, -OC 1-6 Alkyl, -SC 1-6 Alkyl, C 3-7 Carbon ring, -OC 3-7 Carbon rings, 3- to 7-membered heterocycles, or -P(=O)R q1 R q2 The alkyl, alkenyl, alkynyl, carbocyclic, or heterocyclic group is optionally surrounded by 1 to 4 R groups. k replace;

[0047] In some implementation schemes, R q R B Each is independently selected from H, deuterium, =O, halogen, CN, OH, NH2, NHC 1-4 Alkyl, N(C) 1-4 Alkyl)2, C 1-4 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl group, -OC 1-4 Alkyl, -SC 1-4 Alkyl, C 3-7 Carbon ring, -OC 3-7 Carbon rings, 3- to 7-membered heterocycles, or -P(=O)R q1 R q2The alkyl, alkenyl, alkynyl, carbocyclic, or heterocyclic group is optionally surrounded by 1 to 4 R groups. k replace;

[0048] In some implementation schemes, R q R B Each of the following groups is independently selected from H, deuterium, =O, F, Cl, Br, cyano, methyl, ethyl, isopropyl, methoxy, ethoxy, isopropyloxy, methylthio, cyclopropyl, cyclopropyl-O-, cyclobutyl, vinyl, ethynyl, -P(=O)(CH3)2, -P(=O)(CH2CH3)2, -P(=O)(CH3)(cyclopropyl), wherein the methyl, ethyl, isopropyl, methoxy, ethoxy, isopropyloxy, methylthio, cyclopropyl, cyclobutyl, vinyl, and ethynyl groups are optionally prefixed with 1 to 4 R groups. k replace;

[0049] In some implementation schemes, R q R B Each group is independently selected from H, =O, F, Cl, Br, cyano, CH2F, CHF2, CF3, -OCH2F, -OCHF2, -OCF3, -OCD3, methyl, -S-methyl, -S-CF3, ethyl, isopropyl, ethynyl, methoxy, ethoxy, isopropyloxy, propyloxy, cyclopropyl, -O-cyclopropyl, -P(=O)(CH3)2, -P(=O)(CH2CH3)2, -P(=O)(CH3)(cyclopropyl), wherein the methyl, ethyl, isopropyl, ethynyl, methoxy, ethoxy, isopropyloxy, propyloxy, and cyclopropyl groups are optionally prefixed with 1 to 4 R groups. k replace;

[0050] In some implementation schemes, R k Each is independently selected from deuterium, =O, halogen, CN, OH, NH2, -C(=O)NH2, NHC 1-6 Alkyl, N(C) 1-6 Alkyl)2, C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, -OC 1-6 Alkyl, -SC 1-6 Alkyl, -OC 3-6 Carbocyclic rings, -O-3 to 7-membered heterocycles, -NH-C 3-6 Carbon rings, -NH-3 to 7-membered heterocycles, -C 1-4 Alkylene-C 3-6 Carbon ring, -C 1-4 Alkylene-3 to 7-membered heterocycles, C 3-6 The carbon ring, 3 to 7-membered heterocycle, wherein the alkyl, alkylene, alkenyl, alkynyl, carbon ring, or heterocycle is optionally selected from deuterium, halogen, =O, CN, OH, NH2, C1-6 Alkyl, C 1-6 Substituents of alkoxy groups;

[0051] In some implementation schemes, R k Each is independently selected from deuterium, =O, halogen, CN, OH, NH2, -C(=O)NH2, NHC 1-4 Alkyl, N(C) 1-4 Alkyl)2, C 1-4 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl group, -OC 1-4 Alkyl, -SC 1-4 Alkyl, -OC 3-6 Carbocyclic rings, -O-3 to 7-membered heterocycles, C 3-6 Carbocyclic rings, 3 to 7-membered heterocycles, wherein the alkyl, alkenyl, alkynyl, carbocyclic, or heterocycle is optionally selected from 1 to 4 elements selected from deuterium, halogens, CN, OH, NH2, C. 1-4 Alkyl, C 1-4 Substituents of alkoxy groups;

[0052] In some implementation schemes, R k Each is independently selected from deuterium, =O, F, Cl, Br, I, CN, OH, NH2, -C(=O)NH2, NH(CH3), NH(CH2CH3), N(CH3)2, N(CH2CH3)2, methyl, ethyl, vinyl, ethynyl, methoxy, ethoxy, methylthio, -O-cyclopropyl, -NH-cyclopropyl, -CH2-cyclopropyl, -CH2-cyclobutyl, -CH2-cyclopentyl -CH2-cyclohexyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, wherein the methyl, ethyl, vinyl, ethynyl, methoxy, ethoxy, methylthio, -O-cyclopropyl, -NH-cyclopropyl, -CH2-cyclopropyl, -CH2-cyclobutyl, -CH2-cyclopentyl, -CH2-cyclohexyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, optionally selected from 1 to 4 elements selected from deuterium, halogen, CN, OH, NH2, C 1-4 Alkyl, C 1-4 Substituents of alkoxy groups;

[0053] In some implementation schemes, R k Each is independently selected from deuterium, F, Cl, Br, I, CN, OH, NH2, -C(=O)NH2, -CH2OH, methyl, ethyl, vinyl, ethynyl, methoxy, ethoxy, methylthio, -O-cyclopropyl, -NH-cyclopropyl, -CH2-cyclopropyl, -CH2-cyclobutyl, -CH2-cyclopentyl, -CH2-cyclohexyl, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

[0054] As a first embodiment of the present invention, the compound represented by general formula (I) or general formula (I-1) or its stereoisomers, tautomers, racemates, or pharmaceutically acceptable salts thereof are used.

[0055] X is selected from O, S, -S(O)- or -S(O)2-;

[0056] Q is selected from

[0057] Q1 is selected from C 6-10 Aryl, 5- to 10-membered heteroaryl, C 5-10 Carbon rings, 5- to 10-membered heterocycles or The aryl, heteroaryl, carbocyclic, or heterocyclic group is optionally surrounded by 1 to 5 R groups. q replace;

[0058] R is selected from C 1-6 alkyl, -C(=O)R q1 -P(=O)R q1 R q2 -C 1-4 Alkylene-OP(=O)R q1 R q2 -C(O)OC 1-4 Alkylene-OP(=O)R q1 R q2 -S(=O)2R q1 C 3-6 Carbocyclic, 4- to 7-membered heterocyclic, wherein the alkyl, alkylene, carbocyclic or heterocyclic is optionally surrounded by 1 to 4 R... k replace;

[0059] R q1 R q2 Each is independently selected from H, OH, and C. 1-6 Alkyl, C 1-6 Alkoxy, C 3-6 Carbon ring, -OC 3-6 Carbocyclic, 4- to 7-membered heterocyclic, wherein the alkyl, alkoxy, carbocyclic or heterocyclic is optionally surrounded by 1 to 4 R... k replace;

[0060] As an option, R q1 R q2 Direct connection forms a 4- to 7-membered heterocycle, wherein the heterocycle is optionally bounded by 1 to 4 R... k replace;

[0061] R q3 R q4 Each is independently selected from H or arbitrarily selected by 1 to 4 Rs. k One of the following groups is substituted: C1-6 Alkyl, -C 1-6 Alkylene -NH2, -C 1-6 Alkylene -C(=O)NH2, -C 1-6 Alkylene -NHC(=NH)NH2, -C 1-6 Alkylene -NHC(=O)NH2、-C 1-6 alkylene -OH, -C 1-6 Alkylene -SH, -C 1-6 Alkylene -CO2H, -C 1-6 Alkylene-C 3-10 Carbon ring, C 3-10 Carbon ring, -C 1-6 Alkylene-3 to 10-membered heterocycles, 3 to 10-membered heterocycles;

[0062] As an option, R q3 R q4 Direct connection forms C 3-6 Carbon rings or 4- to 7-membered heterocycles, wherein the carbon rings or heterocycles are optionally surrounded by 1 to 4 R... k replace;

[0063] B is selected from C. 6-10 Aryl, 5- to 10-membered heteroaryl, C 5-10 Carbocyclic rings, 5- to 10-membered heterocyclic rings, wherein the aryl, heteroaryl, carbocyclic, or heterocyclic rings are optionally surrounded by 1 to 5 R groups. B replace;

[0064] R 1 R 2 R 3 R 4 R 5 Each is independently selected from H, halogens, CN, OH, and C. 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, -OC 1- 6-alkyl, -SC 1-6 Alkyl, C 3-6 Carbocyclic, 3- to 7-membered heterocyclic, wherein the alkyl, alkenyl, alkynyl, carbocyclic or heterocyclic is optionally surrounded by 1 to 4 R... k replace;

[0065] As an option, R 1 R 4 Direct connection forms C 3-6 Carbon rings or 4- to 7-membered heterocycles, wherein the carbon rings or heterocycles are optionally surrounded by 1 to 6 R... k replace;

[0066] R q R B Each is independently selected from H, deuterium, =O, halogen, CN, OH, NH2, NHC1-6 Alkyl, N(C) 1-6 Alkyl)2, C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, -OC 1-6 Alkyl, -SC 1-6 Alkyl, C 3-7 Carbon ring, -OC 3-7 Carbon rings, 3- to 7-membered heterocycles, or -P(=O)R q1 R q2 The alkyl, alkenyl, alkynyl, carbocyclic, or heterocyclic group is optionally surrounded by 1 to 4 R groups. k replace;

[0067] R k Each is independently selected from deuterium, =O, halogen, CN, OH, NH2, -C(=O)NH2, NHC 1-6 Alkyl, N(C) 1-6 Alkyl)2, C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, -OC 1-6 Alkyl, -SC 1-6 Alkyl, -OC 3-6 Carbocyclic rings, -O-3 to 7-membered heterocycles, -NH-C 3- 6-membered carbon rings, -NH-3 to 7-membered heterocycles, -C 1-4 Alkylene-C 3-6 Carbon ring, -C 1-4 Alkylene-3 to 7-membered heterocycles, C 3-6 The carbon ring, 3 to 7-membered heterocycle, wherein the alkyl, alkylene, alkenyl, alkynyl, carbon ring, or heterocycle is optionally selected from deuterium, halogen, =O, CN, OH, NH2, C 1-6 Alkyl, C 1-6 The alkoxy group is replaced by a substituent.

[0068] As a second embodiment of the present invention, the compound represented by general formula (I) or general formula (I-1) or its stereoisomers, tautomers, racemates, or pharmaceutically acceptable salts thereof...

[0069] R is selected from C 1-4 alkyl, -C(=O)R q1 -P(=O)R q1 R q2 -C 1-2 Alkylene-OP(=O)R q1 R q2 -C(O)OC 1-2 Alkylene-OP(=O)R q1 Rq2 -S(=O)2R q1 C 3-6 Carbocyclic, 4- to 7-membered heterocyclic, wherein the alkyl, alkylene, carbocyclic or heterocyclic is optionally surrounded by 1 to 4 R... k replace;

[0070] R q1 R q2 Each is independently selected from H, OH, and C. 1-4 Alkyl, C 1-4 Alkoxy, C 3-6 Carbon ring, -OC 3-6 Carbocyclic, 4- to 7-membered heterocyclic, wherein the alkyl, alkoxy, carbocyclic or heterocyclic is optionally surrounded by 1 to 4 R... k replace;

[0071] As an option, R q1 R q2 Direct connection forms a 4- to 7-membered heterocycle, wherein the heterocycle is optionally bounded by 1 to 4 R... k replace;

[0072] R q3 R q4 Each is independently selected from H or arbitrarily selected by 1 to 4 Rs. k One of the following groups is substituted: C 1-4 Alkyl, -C 1-4 Alkylene -NH2, -C 1-4 Alkylene -C(=O)NH2, -C 1-4 Alkylene -NHC(=NH)NH2, -C 1-4 Alkylene -NHC(=O)NH2、-C 1-4 alkylene -OH, -C 1-4 Alkylene -SH, -C 1-4 Alkylene -CO2H, -C 1-4 Alkylene-C 6-10 Aryl, C 6-10 Aryl, -C 1-4 alkylene-5 to 10-membered heteroaryl, 5 to 10-membered heteroaryl, C 3-6 cycloalkyl, 4- to 7-membered heterocyclic groups, -CH2-C 3-6 Cycloalkyl groups, -CH2-4 to 7-membered heterocyclic groups;

[0073] As an option, R q3 R q4 Direct connection forms C 3-6 Cycloalkyl or 4- to 7-membered heterocycloalkyl, wherein the cycloalkyl or heterocycloalkyl is optionally surrounded by 1 to 4 R... k replace;

[0074] Q1 is selected from phenyl, benzo[C] 4-6Carbocyclic, benzo[4-6] heterocyclic, 5-6 fused heteroaryl, 5-6 fused heterocyclic, 8-10 fused heteroaryl or Q1 is arbitrarily selected by 1 to 4 Rs q replace;

[0075] B is selected from phenyl, benzo[C] 4-6 Carbocyclic, benzo[4- to 6-membered heterocyclic, 5- to 6-membered heteroaryl, 8- to 10-membered fused-ring heteroaryl, wherein B is optionally surrounded by 1 to 4 R[]. B replace;

[0076] R 1 R 2 R 3 R 4 R 5 Each is independently selected from H, halogens, CN, OH, and C. 1-4 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl group, -OC 1-4 Alkyl, -SC 1-4 Alkyl, C 3-6 Carbocyclic, 3- to 7-membered heterocyclic, wherein the alkyl, alkenyl, alkynyl, carbocyclic or heterocyclic is optionally surrounded by 1 to 4 R... k replace;

[0077] As an option, R 1 R 4 Direct connection forms C 3-6 Carbon rings or 4- to 7-membered heterocycles, wherein the carbon rings or heterocycles are optionally surrounded by 1 to 4 R... k replace;

[0078] R q R B Each is independently selected from H, deuterium, =O, halogen, CN, OH, NH2, NHC 1-4 Alkyl, N(C) 1-4 Alkyl)2, C 1-4 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl group, -OC 1-4 Alkyl, -SC 1-4 Alkyl, C 3-7 Carbon ring, -OC 3-7 Carbon rings, 3- to 7-membered heterocycles, or -P(=O)R q1 R q2 The alkyl, alkenyl, alkynyl, carbocyclic, or heterocyclic group is optionally surrounded by 1 to 4 R groups. k replace;

[0079] R k Each is independently selected from deuterium, =O, halogen, CN, OH, NH2, -C(=O)NH2, NHC 1-4Alkyl, N(C) 1-4 Alkyl)2, C 1-4 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl group, -OC 1-4 Alkyl, -SC 1-4 Alkyl, -OC 3-6 Carbocyclic rings, -O-3 to 7-membered heterocycles, C 3-6 Carbocyclic rings, 3 to 7-membered heterocycles, wherein the alkyl, alkenyl, alkynyl, carbocyclic, or heterocycle is optionally selected from 1 to 4 elements selected from deuterium, halogens, CN, OH, NH2, C. 1-4 Alkyl, C 1-4 Substituents of alkoxy groups;

[0080] The definitions of the remaining functional groups are the same as those in the first embodiment of the present invention.

[0081] As a third embodiment of the present invention, the compound represented by general formula (I) or general formula (I-1) above, or its stereoisomers, tautomers, racemates, or pharmaceutically acceptable salts,

[0082] R is selected from -C(=O)R q1 -P(=O)R q1 R q2 ,-methylene-OP(=O)R q1 R q2 -C(O)O-methylene-OP(=O)R q1 R q2 -S(=O)2R q1 C 3-6 Carbocyclic rings, 4- to 7-membered heterocycles, wherein the methylene group, carbocyclic ring, or heterocycle is optionally surrounded by 1 to 4 R groups. k replace;

[0083] R q1 R q2 Each is independently selected from H, OH, or arbitrarily selected by 1 to 4 R. k The substitution is made with one of the following groups: methyl, ethyl, propyl, butyl, isopropyl, tert-butyl, methoxy, ethoxy, isopropyloxy, tert-butyloxy, cyclopropyl, cyclobutyl, cyclopentyl, oxecyclobutyl, oxecyclopentyl, oxecyclohexyl, azircyclobutyl, azircyclopentyl, azircyclohexyl;

[0084] R q3 R q4 Each is independently selected from H or arbitrarily selected by 1 to 4 Rs. kThe substitution is performed using one of the following groups: methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, -CH2NH2, -CH2CH2NH2, -CH2CH2CH2NH2, -CH2CH2CH2CH2NH2, -CH2NHC(=NH)NH2, -CH2CH2NHC(=NH)NH2, -CH2CH2CH2NHC(=NH)NH2, -CH2CH2CH2NHC(=O)NH2, -CH2OH, -CH2SH, -CH2CH2SH, -CH2CO2H, -CH2 CH2CO2H, -CH2C(=O)NH2, -CH2CH2C(=O)NH2, -CH2CH2CH2C(=O)NH2, -CH2CH2CH2CH2C(=O)NH2, -CH2-phenyl, -CH2-indole, -CH2-5 to 10-membered heteroaryl, -CH2-C 3-6 cycloalkyl, C 3- 6-membered cycloalkyl, 4- to 7-membered heterocyclic alkyl, or -CH2-4- to 7-membered heterocyclic alkyl;

[0085] R 1 R 2 R 3 R 4 R 5 Each of the following groups is independently selected from H, F, Cl, Br, cyano, methyl, ethyl, methoxy, ethoxy, methylthio, cyclopropyl, cyclobutyl, vinyl, and ethynyl, wherein the methyl, ethyl, methoxy, ethoxy, methylthio, cyclopropyl, cyclobutyl, vinyl, and ethynyl groups are optionally prefixed with 1 to 4 R groups. k replace;

[0086] As an option, R 1 R 4 Direct connection forms C 3-6 Carbon ring, wherein the carbon ring is optionally divided by 1 to 4 R k replace;

[0087] B is selected from Or phenyl, wherein the B is optionally surrounded by 1 to 4 R B replace;

[0088] R q R BEach of the following groups is independently selected from H, deuterium, =O, F, Cl, Br, cyano, methyl, ethyl, isopropyl, methoxy, ethoxy, isopropyloxy, methylthio, cyclopropyl, cyclopropyl-O-, cyclobutyl, vinyl, ethynyl, -P(=O)(CH3)2, -P(=O)(CH2CH3)2, -P(=O)(CH3)(cyclopropyl), wherein the methyl, ethyl, isopropyl, methoxy, ethoxy, isopropyloxy, methylthio, cyclopropyl, cyclobutyl, vinyl, and ethynyl groups are optionally prefixed with 1 to 4 R groups. k replace;

[0089] R k Each is independently selected from deuterium, =O, F, Cl, Br, I, CN, OH, NH2, -C(=O)NH2, NH(CH3), NH(CH2CH3), N(CH3)2, N(CH2CH3)2, methyl, ethyl, vinyl, ethynyl, methoxy, ethoxy, methylthio, -O-cyclopropyl, -NH-cyclopropyl, -CH2-cyclopropyl, -CH2-cyclobutyl, -CH2-cyclopentyl -CH2-cyclohexyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, wherein the methyl, ethyl, vinyl, ethynyl, methoxy, ethoxy, methylthio, -O-cyclopropyl, -NH-cyclopropyl, -CH2-cyclopropyl, -CH2-cyclobutyl, -CH2-cyclopentyl, -CH2-cyclohexyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, optionally selected from 1 to 4 elements selected from deuterium, halogen, CN, OH, NH2, C 1-4 Alkyl, C 1-4 Substituents of alkoxy groups;

[0090] The definitions of the remaining functional groups are the same as those in the first or second embodiment of the present invention.

[0091] As a fourth embodiment of the present invention, the compound represented by general formula (I) or general formula (I-1) above, or its stereoisomers, tautomers, racemates, or pharmaceutically acceptable salts,

[0092] R is selected from -C(=O)R q1 -P(=O)R q1 R q2 -CH2-OP(=O)R q1 R q2 -C(O)O-CH2-OP(=O)R q1 R q2 -S(=O)2R q1 The CH2 is optionally converted by 1 to 2 R k replace;

[0093] R q3 R q4Each is independently selected from H or arbitrarily selected by 1 to 4 Rs. k The substitution is made with one of the following groups: methyl, ethyl, propyl, butyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, -CH2NH2, -CH2CH2NH2, -CH2CH2CH2NH2, -CH2CH2CH2CH2NH2, -CH2NHC(=NH)NH2, -CH2CH2NHC(=NH)NH2, -CH2CH2CH2NHC(=NH)NH2, -CH2CH2CH2NHC(=O)NH2, -CH2OH, -CH2SH, -CH2CH2SH, -CH2CO2H, -CH2 CH2CO2H, -CH2-phenyl, -CH2-imidazolyl, -CH2-indolyl, cyclopropyl, cyclobutyl, cyclopentyl, oxecyclobutyl, oxecyclopentyl, oxecyclohexyl, azircyclobutyl, azircyclopentyl, azircyclohexyl, -CH2-cyclopropyl, -CH2-cyclobutyl, -CH2-cyclopentyl, -CH2-oxecyclobutyl, -CH2-oxecyclopentyl, -CH2-azircyclobutyl, -CH2-azircyclopentyl, -CH2-azircyclohexyl;

[0094] As an option, R q3 R q4 Directly linked to form cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, oxacyclobutyl, oxacyclopentyl, oxacyclohexyl, aziridine, aziridinepentyl, and aziridinehexyl, wherein the cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, oxacyclobutyl, oxacyclopentyl, oxacyclohexyl, aziridine, aziridinepentyl, and aziridinehexyl are optionally separated by 1 to 4 R... k replace;

[0095] Q1 is selected from Q1 is arbitrarily selected by 1 to 4 Rs q replace;

[0096] R 1 R 2 R 3 R 4 R 5 Each is independently selected from H, methyl, ethyl, CH2F, CHF2, CF3;

[0097] R q R BEach group is independently selected from H, =O, F, Cl, Br, cyano, CH2F, CHF2, CF3, -OCH2F, -OCHF2, -OCF3, -OCD3, methyl, -S-methyl, -S-CF3, ethyl, isopropyl, ethynyl, methoxy, ethoxy, isopropyloxy, propyloxy, cyclopropyl, -O-cyclopropyl, -P(=O)(CH3)2, -P(=O)(CH2CH3)2, -P(=O)(CH3)(cyclopropyl), wherein the methyl, ethyl, isopropyl, ethynyl, methoxy, ethoxy, isopropyloxy, propyloxy, and cyclopropyl groups are optionally prefixed with 1 to 4 R groups. k replace;

[0098] R k Each is independently selected from deuterium, F, Cl, Br, I, CN, OH, NH2, -C(=O)NH2, -CH2OH, methyl, ethyl, vinyl, ethynyl, methoxy, ethoxy, methylthio, -O-cyclopropyl, -NH-cyclopropyl, -CH2-cyclopropyl, -CH2-cyclobutyl, -CH2-cyclopentyl, -CH2-cyclohexyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl;

[0099] The definitions of the remaining functional groups are the same as those in the first, second, or third embodiments of the present invention.

[0100] As a fifth embodiment of the present invention, the compound represented by general formula (I) or general formula (I-1) above, or its stereoisomers, tautomers, racemates, or pharmaceutically acceptable salts,

[0101] Q1 is selected from Q1 is arbitrarily selected by 1 to 3 Rs q replace;

[0102] B is selected from

[0103] The definitions of the remaining functional groups are the same as those in the first, second, third, or fourth embodiments of the present invention.

[0104] As a sixth embodiment of the present invention, the compound represented by general formula (I) or general formula (I-1) above, or its stereoisomers, tautomers, racemates, or pharmaceutically acceptable salts,

[0105] X is selected from O and S;

[0106] R is selected from Preferably, R is selected from

[0107] B is selected from

[0108] The definitions of the remaining functional groups are the same as those in the first, second, third, fourth, or fifth embodiments of the present invention.

[0109] As a seventh embodiment of the present invention, the compound represented by the above general formula (I) or its stereoisomers, tautomers, racemates, or pharmaceutically acceptable salts are used.

[0110] The compounds represented by general formula (I) are selected from general formula (Ib).

[0111] R 4 Selected from H or C 1-4 Alkyl, preferably H or methyl;

[0112] R b Each is independently selected from deuterium, halogens, CN, and C. 1-6 Alkyl, -OC 1-6 Alkyl group, wherein the alkyl group is optionally substituted with 1 to 4 substituents selected from deuterium, halogen, cyclopropyl, preferably F, Cl, Br, methyl, ethyl, methoxy, ethoxy, CH2F, CHF2, CF3, -OCH2F, -OCHF2, -OCF3, -OCD3, -OCH2D, -OCHD2;

[0113] R q Each is independently selected from deuterium, halogens, CN, and C. 1-6 Alkyl, -OC 1-6 Alkyl group, wherein the alkyl group is optionally substituted with 1 to 4 substituents selected from deuterium, halogen, cyclopropyl, preferably F, Cl, or Br;

[0114] R is selected from -P(=O)(OH)2;

[0115] n1 and n2 are selected from 0, 1, 2, and 3.

[0116] This invention relates to compounds or their stereoisomers, tautomers, racemates, or pharmaceutically acceptable salts as shown below, wherein the compound is selected from one of the structures shown in Table E.

[0117] Table E

[0118] The compound of the present invention has a pharmaceutically acceptable salt, wherein the salt is selected from acid or base molecules, wherein the base molecule is selected from at least one of lithium salt, sodium salt, potassium salt, calcium salt, magnesium salt, aluminum salt, iron salt, zinc salt, ammonium salt, lysine salt, arginine salt, L-arginine salt, histidine salt, L-histidine salt, meglumine salt, dimethylglucosamine salt, ethylglucosamine salt, dicyclohexylamine salt, 1,6-hexanediamine salt, glucosamine salt, triethanolamine salt, sarcosine salt, serine salt, trihydroxymethylaminomethane salt, aminopropylene glycol salt, 1-amino-2,3,4-butanetriol salt, L-lysine salt, ornithine salt, or choline salt, preferably meglumine salt; wherein the acid molecule is selected from at least one of hydrochloric acid, sulfuric acid, hydrobromic acid, phosphoric acid, citric acid, oxalic acid, maleic acid, fumaric acid, L-tartaric acid, and p-toluenesulfonic acid, preferably hydrochloric acid.

[0119] The compounds of this invention are pharmaceutically acceptable salts, wherein the chemical molar ratio of the compounds to acid or base molecules is 1:2 to 1:1 or 1:1 to 2:1.

[0120] The compounds of this invention are pharmaceutically acceptable salts of which have a stoichiometric ratio of 1:2, 1:1, or 2:1 with an acid or base molecule.

[0121] This invention relates to pharmaceutically acceptable salts of the aforementioned compounds, wherein the salts are selected from base molecules.

[0122] The chemical molar ratio of the compound to the base molecule described in this invention is 1:2 to 1:1.

[0123] The chemical molar ratio of the compound to the base molecule described in this invention is 1:2 or 1:1.

[0124] The chemical molar ratio of the compound described in this invention to meglumine is 1:2.

[0125] The compound of this invention has a pharmaceutically acceptable salt, wherein the salt is...

[0126] The salt is preferably amorphous, and the X-ray diffraction pattern of the amorphous salt has no obvious characteristic peaks; the preferred X-ray diffraction pattern is shown in Figure 1.

[0127] This invention relates to a pharmaceutical composition comprising the above-described compound or its stereoisomers, tautomers, racemates, pharmaceutically acceptable salts, and pharmaceutically acceptable carriers.

[0128] This invention relates to the use of the above-mentioned compounds or their stereoisomers, tautomers, racemates, pharmaceutically acceptable salts, or the above-mentioned pharmaceutical compositions in the preparation of medicaments for inhibiting Nav1.8-related diseases.

[0129] This invention relates to the use of the above-mentioned compounds or their stereoisomers, tautomers, racemates, pharmaceutically acceptable salts, or the above-mentioned pharmaceutical compositions in the preparation of medicaments for treating and / or alleviating pain or pain-related diseases, multiple sclerosis, Sharma-Tutankhamun syndrome, incontinence, pathological cough, or arrhythmia; preferably, the pain is selected from chronic pain, acute pain, inflammatory pain, cancer pain, postoperative pain, neuropathic pain, musculoskeletal pain, primary pain, intestinal pain, and idiopathic pain; the postoperative pain is preferably selected from pain after bunion removal, hernia repair, and abdominoplasty.

[0130] This invention relates to a pharmaceutical composition or pharmaceutical formulation comprising a therapeutically effective amount of the compound of the invention or its stereoisomers, tautomers, racemates, pharmaceutically acceptable salts, and pharmaceutical excipients. The pharmaceutical composition may be in unit dosage form (the amount of the active ingredient in a unit dosage form is also referred to as a "dosage strength").

[0131] The present invention also provides a method for treating diseases in mammals, comprising administering to the mammal a therapeutically effective amount of the compound of the present invention or its stereoisomers, tautomers, racemates, pharmaceutically acceptable salts, or pharmaceutical compositions. In some embodiments, the mammals described in the present invention include humans.

[0132] The term "effective amount" or "therapeutic effective amount" as used in this application means that administering a sufficient amount of the compound disclosed in this application will alleviate, to some extent, one or more symptoms of the disease or condition being treated. In some embodiments, the result is a reduction and / or mitigation of the signs, symptoms, or causes of the disease, or any other desired alteration of the biological system. For example, an "effective amount" for therapeutic use is the amount of the compound disclosed in this application required to provide a clinically significant reduction in disease symptoms.Examples of therapeutically effective doses include, but are not limited to, 1-1500 mg, 1-1000 mg, 1-800 mg, 1-600 mg, 2-600 mg, 3-600 mg, 4-600 mg, 5-600 mg, 6-600 mg, 10-600 mg, 20-600 mg, 25-600 mg, 30-600 mg, 40-600 mg, 50-600 mg, 60-600 mg, 70-600 mg, 75-600 mg, 80-600 mg, 90-600 mg, 100-600 mg, 200-600 mg, 1-500 mg, 2-500 mg, 3-500 mg. g, 4-500mg, 5-500mg, 6-500mg, 10-500mg, 20-500mg, 25-500mg, 30-500mg, 40-500mg, 50-500mg, 60-500mg, 70-500mg, 75-500mg, 80-500mg , 90-500mg, 100-500mg, 125-500mg, 150-500mg, 200-500mg, 250-500mg, 300-500mg, 400-500mg, 5-400mg, 10-400mg, 20-400mg, 25-400mg, 30-400mg, 40-400mg, 50-400mg, 60-400mg, 70-400mg, 75-400mg, 80-400mg, 90-400mg, 100-400mg, 125-400mg, 150-400mg, 200-400mg, 25 0-400mg, 300-400mg, 1-300mg, 2-300mg, 5-300mg, 10-300mg, 20-300mg, 25-300mg, 30-300mg, 40-300mg, 50-300mg, 60-300mg, 70-300mg, 7 5-300mg, 80-300mg, 90-300mg, 100-300mg, 125-300mg, 150-300mg, 200-300mg, 250-300mg, 1-200mg, 2-200mg, 5-200mg, 10-200mg, 20-200 mg, 25-200mg, 30-200mg, 40-200mg, 50-200mg, 60-200mg, 70-200mg, 75-200mg, 80-200mg, 90-200mg, 100-200mg, 125-200mg, 150-200mg;.

[0133] In some embodiments, the pharmaceutical composition includes, but is not limited to, 1-1500 mg, 1-1000 mg, 1-800 mg, 1-600 mg, 20-400 mg, 25-200 mg, 1 mg, 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 55 mg, 65 mg, 70 mg, 75 mg, 80 mg, 85 mg, 90 mg, 95 mg, 100 mg, 110 mg, 120 mg, 125 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, 190 mg, 200 mg, 210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 300 mg of the compound of the present invention or its stereoisomers, tautomers, or pharmaceutically acceptable salts.

[0134] A method for treating a disease in a mammal, the method comprising administering to a subject a therapeutically effective amount of the compound of the present invention or its stereoisomers, tautomers, racemates, or pharmaceutically acceptable salts, preferably 1-1500 mg, wherein the disease is preferably pain.

[0135] A method for treating a disease in mammals, the method comprising administering a drug, a compound of the present invention or its stereoisomers, tautomers, racemates, or pharmaceutically acceptable salts thereof, to a subject at a daily dose of 1-1500 mg / day, said daily dose being a single dose or multiple doses, and in some embodiments, the daily dose including but not limited to 10-1500 mg / day, 10-1000 mg / day, 10-800 mg / day, 25-800 mg / day, 50-800 mg / day, etc. 100-800mg / day, 200-800mg / day, 25-400mg / day, 50-400mg / day, 100-400mg / day, 200-400mg / day. In some embodiments, the daily dose includes, but is not limited to, 10mg / day, 20mg / day, 25mg / day, 50mg / day, 100mg / day, 125mg / day, 150mg / day, 200mg / day, 400mg / day, 600mg / day, and 800mg / day.

[0136] The diseases described in this invention include pain or pain-related diseases, multiple sclerosis, Sharma-Tutankhamun syndrome, incontinence, pathological cough, or arrhythmia. Preferably, the pain is selected from chronic pain, acute pain, inflammatory pain, cancer pain, postoperative pain, neuropathic pain, musculoskeletal pain, primary pain, intestinal pain, and idiopathic pain; the postoperative pain is preferably selected from pain from bunion removal surgery, hernia repair surgery, and abdominoplasty.

[0137] This invention relates to a kit that may include a single-dose or multi-dose composition comprising a compound of the present invention or its stereoisomers, tautomers, racemates, or pharmaceutically acceptable salts thereof, wherein the amount of the compound of the present invention or its stereoisomers, tautomers, racemates, or pharmaceutically acceptable salts is the same as the amount in the aforementioned pharmaceutical composition.

[0138] The compounds of the present invention also include their deuterated derivatives, solvates, prodrugs, metabolites, and cocrystals.

[0139] In this invention, the amount of the compound of the invention or its stereoisomers, tautomers, racemates, or pharmaceutically acceptable salts is converted in each case as a free base.

[0140] "Product specification" refers to the weight of the active pharmaceutical ingredient contained in each vial, tablet, or other unit of preparation.

[0141] Unless otherwise stated, the terms used in the specification and claims have the following meanings.

[0142] The carbon, hydrogen, oxygen, sulfur, nitrogen, phosphorus, F, Cl, Br, I, etc. involved in the groups and compounds described in this invention include their isotopic forms. That is, the carbon, hydrogen, oxygen, sulfur, nitrogen, phosphorus, F, Cl, Br, I, etc. involved in the groups and compounds described in this invention may be optionally further replaced by one or more of their corresponding isotopes, wherein the isotopes of carbon include 11 C 12 C 13 C and 14 C, the isotopes of hydrogen include protium (H), deuterium (D, also called heavy hydrogen), and tritium (T, also called superheavy hydrogen), and the isotopes of oxygen include 15 O、 16 O、 17 O and 18 O, isotopes of sulfur include 32 S, 33 S, 34 S, 35 S and 36 S, nitrogen isotopes include 13 N、 14 N and 15 N, isotopes of fluorine include 17 F, 18 F and 19 F, isotopes of chlorine include 35 Cl、 36 Cl and 37 Cl, isotopes of bromine include 79 Br and 81 Br, an isotope of iodine, includes 123 I, 125I, phosphorus isotopes include 31 P, 32 P. In some embodiments, at the position of the deuterium substituent, the deuterium isotope abundance is greater than the natural deuterium isotope abundance (0.015%), preferably greater than 50%, more preferably greater than 60%, greater than 70%, greater than 80%, greater than 90%, greater than 95%, greater than 96%, greater than 97%, greater than 98%, greater than 99%, greater than 99.5%, or 100%. In some cases, such as when “hydrogen” and “deuterium” appear as parallel terms, or when “hydrogen” is replaced by “deuterium”, the term “hydrogen” refers to the isotope of hydrogen. 1 "H", while "deuterium" represents the isotope of hydrogen. 2 H”; or it should be understood that at this position in the compound, hydrogen, existing in its natural abundance at various isotopes at that position, is replaced by deuterium, existing in its abundance at a level greater than that of the natural deuterium isotopes (e.g., deuterium abundance greater than 50%, greater than 60%, greater than 70%, greater than 80%, greater than 90%, greater than 95%, greater than 96%, greater than 97%, greater than 98%, greater than 99%, greater than 99.5%, or 100%).

[0143] “CN” refers to cyano.

[0144] "Halogen" refers to F, Cl, Br or I.

[0145] "Halogen-substituted" refers to substitution with F, Cl, Br, or I, including but not limited to 1 to 10 substituents selected from F, Cl, Br, or I, 1 to 6 substituents selected from F, Cl, Br, or I, and 1 to 4 substituents selected from F, Cl, Br, or I. "Halogen-substituted" is abbreviated as "halogenated".

[0146] "alkyl" refers to a substituted or unsubstituted straight-chain or branched saturated aliphatic hydrocarbon group, including but not limited to alkyl groups with 1 to 20 carbon atoms, alkyl groups with 1 to 8 carbon atoms, alkyl groups with 1 to 6 carbon atoms, and alkyl groups with 1 to 4 carbon atoms. Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, neobutyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, and their various branched isomers; the alkyl group can be monovalent, divalent, trivalent, or tetravalent.

[0147] "Alkylene" refers to substituted or unsubstituted straight-chain and branched divalent saturated hydrocarbon groups, including -(CH2). v - (v is an integer from 1 to 10), alkylene examples include, but are not limited to, methylene, ethylene, propylene, and butylene.

[0148] "Cycloalkyl" refers to a substituted or unsubstituted saturated carbocyclic hydrocarbon group, typically having 3 to 12 carbon atoms. Cycloalkyl groups can be monocyclic, fused, bridged, or spirocyclic. Non-limiting examples include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclobutyl-cyclobutyl, cyclobutyl-spirobutyl, adamantane, etc. Cycloalkyl groups can be monovalent, divalent, trivalent, or tetravalent.

[0149] "Heterocyclic alkyl" refers to a substituted or unsubstituted saturated cyclic hydrocarbon group containing heteroatoms, including but not limited to 3 to 12 atoms or 3 to 8 atoms, containing 1 to 3 heteroatoms selected from N, O, S, or Se. The C, N, and S atoms on the ring of the heterocyclic alkyl group can be oxidized to various oxidation states. Heterocyclic alkyl groups can be monocyclic, fused, bridged, or spirocyclic. Heterocyclic alkyl groups can be attached to heteroatoms or carbon atoms. Non-limiting examples include epoxyethyl, aziridinepropyl, oxacyclobutyl, aziridinebutyl, tetrahydrofuranyl, tetrahydro-2H-pyranyl, dioxopentyl, dioxohexyl, pyrrolylalkyl, piperidinyl, imidazoalkyl, oxazolidinyl, oxazinylalkyl, morpholinyl, hexahydropyrimidinyl, piperazineyl, etc. Heterocyclic alkyl groups can be monovalent, divalent, trivalent, or tetravalent.

[0150] "Alkenyl" refers to a substituted or unsubstituted straight-chain and branched unsaturated hydrocarbon group having at least one, typically one, two, or three, carbon-carbon double bonds. The main chain has, but is not limited to, 2 to 10, 2 to 6, or 2 to 4 carbon atoms. Examples of alkenyl groups include, but are not limited to, vinyl, allyl, 1-propenyl, 2-propenyl, 1-butenyl, 2-butenyl, 3-butenyl, 1-pentenyl, 2-pentenyl, 3-pentenyl, 4-pentenyl, 1-methyl-1-butenyl, 2-methyl-1-butenyl, 2... -Methyl-3-butenyl, 1-hexenyl, 2-hexenyl, 3-hexenyl, 4-hexenyl, 5-hexenyl, 1-methyl-1-pentenyl, 2-methyl-1-pentenyl, 1-heptenyl, 2-heptenyl, 3-heptenyl, 4-heptenyl, 1-octenyl, 3-octenyl, 1-nonenyl, 3-nonenyl, 1-decenyl, 4-decenyl, 1,3-butadiene, 1,3-pentadiene, 1,4-pentadiene, and 1,4-hexadiene, etc.; the alkenyl group can be monovalent, divalent, trivalent, or tetravalent.

[0151] "Alynyl" refers to a substituted or unsubstituted straight-chain and branched unsaturated hydrocarbon group having at least one, typically one, two, or three, carbon-carbon triple bonds. The main chain comprises 2 to 10 carbon atoms, including but not limited to having 2 to 6 carbon atoms on the main chain, or 2 to 4 carbon atoms on the main chain. Examples of alkynyl groups include, but are not limited to, ethynyl, propynyl, 1-propynyl, 2-propynyl, 1-butynyl, 2-butynyl, 3-butynyl, 1-pentynyl, 2-pentynyl, 3-pentynyl, and 4-pentynyl. The alkynyl group can be monovalent, divalent, trivalent, or tetravalent.

[0152] "Alkoxy" refers to a substituted or unsubstituted -O-alkyl group. Non-limiting examples include methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, sec-butoxy, tert-butoxy, n-pentoxy, n-hexoxy, cyclopropoxy, and cyclobutoxy.

[0153] "Carbocyclic group" or "carbocyclic ring" refers to a substituted or unsubstituted aromatic or non-aromatic ring. The aromatic or non-aromatic ring can be a 3- to 8-membered monocyclic ring, a 4- to 12-membered bicyclic ring, a 10- to 15-membered tricyclic ring, or a 12- to 18-membered quaternary system. The carbocyclic group can be attached to an aromatic or non-aromatic ring, and the ring can be optionally a monocyclic, fused, bridged, or spirocyclic ring. Non-limiting examples include cyclopropane, cyclobutane, cyclopentane, cyclohexane, cycloheptane, 1-cyclopentyl-1-enyl, 1-cyclopentyl-2-enyl, 1-cyclopentyl-3-enyl, cyclohexyl, 1-cyclohexyl-2-enyl, 1-cyclohexyl-3-enyl, cyclohexenyl, benzene ring, naphthalene ring, etc. "Carbocyclic group" or "carbon ring" can be monovalent, divalent, trivalent or tetravalent.

[0154] "Heterocyclic group" or "heterocyclic" refers to a substituted or unsubstituted aromatic or non-aromatic ring. The aromatic or non-aromatic ring can be a 3- to 8-membered monocyclic ring, a 4- to 12-membered bicyclic ring, a 10- to 15-membered tricyclic ring, or a 12- to 18-membered quaternary system, and contains one or more (including but not limited to 2, 3, 4 or 5) heteroatoms selected from N, O, S or Se. The C, N, S or Se selectively substituted in the ring of the heterocyclic group can be oxidized to various oxidation states. The heterocyclic group can be attached to a heteroatom or a carbon atom, and can be attached to an aromatic ring or a non-aromatic ring. The heterocyclic group is optionally a monocyclic, bridged, fused, or spirocyclic ring. Non-limiting examples include epoxyethyl, aziridinepropyl, oxacyclobutyl, aziridinebutyl, 1,3-dioxopentyl, 1,4-dioxopentyl, 1,3-dioxahexane, aziridineheptyl, pyridinyl, furanyl, thiophene, pyranyl, N-alkylpyrroleyl, pyrimidinyl, pyrazinyl, pyridazinyl, imidazolyl, piperidinyl, morpholinyl, thiomorpholinyl, 1,3-dithioyl, dihydrofuranyl, dihydropyranyl, dithiapentylcycloyl. Tetrahydrofuranyl, tetrahydropyrrolyl, tetrahydroimidazolyl, tetrahydrothiazolyl, tetrahydropyranyl, benzimidazolyl, benzopyridyl, pyrrolopyridyl, benzodihydrofuranyl, pyrrolyl, pyrazolyl, thiazolyl, oxazolyl, pyrazinyl, indazolyl, benzothiophene, benzofuranyl, benzopyrrolyl, benzimidazolyl, benzothiazolyl, benzooxazolyl, benzopyridyl, benzopyrimidinyl, benzopyrazinyl, piperazinyl, azabicyclo[3.2.1]octyl, azabicyclo[5.2.0]nonyl, oxatricyclo[5.3.1.1]dodecyl, azaadamantyl, oxaspiro[3.3]heptyl, "Heterocyclic group" or "heterocyclic" can be monovalent, divalent, trivalent or tetravalent.

[0155] A "spirocyclic" or "spirocyclic group" refers to a polycyclic group in which substituted or unsubstituted rings share a single atom (called a spiro atom). The number of ring atoms in a spirocyclic system includes, but is not limited to, 5 to 20, 6 to 14, 6 to 12, or 6 to 10. One or more rings may contain zero or more (including but not limited to 1, 2, 3, or 4) double bonds, and optionally, may contain 0 to 5 double bonds selected from N, O, or S (=O). n Heteroatoms (n is 0, 1, or 2). Non-limiting embodiments include:

[0156] "Spirocyclic" or "spirocyclic group" can be monovalent, divalent, trivalent or tetravalent.

[0157] "Circular fused" or "circular fused group" refers to a polycyclic group in which each ring in a system shares a pair of adjacent atoms with other rings in the system. One or more rings may contain zero or more (including but not limited to 1, 2, 3 or 4) double bonds and may be substituted or unsubstituted. Each ring in a circular fused system may contain 0 to 5 heteroatoms or groups containing heteroatoms (including but not limited to those selected from N, S (=O)). n Or O, where n is 0, 1, or 2). The number of ring atoms in a cyclic system includes, but is not limited to, 5 to 20, 5 to 14, 5 to 12, and 5 to 10. Non-limiting examples include: "Cyclone" or "cyclone base" can be monovalent, divalent, trivalent, or tetravalent.

[0158] A “bridged ring” or “bridged ring group” refers to a substituted or unsubstituted polycyclic group containing any two atoms that are not directly connected, and may contain zero or more double bonds. Any ring in a bridged ring system may contain 0 to 5 groups selected from heteroatoms or containing heteroatoms (including but not limited to N, S(=O)n, or O, where n is 0, 1, or 2). The number of ring atoms includes, but is not limited to, 5 to 20, 5 to 14, 5 to 12, or 5 to 10. Non-limiting examples include: Cubicane, adamantane. "Bridged ring" or "bridged ring group" can be monovalent, divalent, trivalent, or tetravalent.

[0159] "Aryl" or "aromatic ring" refers to a substituted or unsubstituted aromatic hydrocarbon group having a monocyclic or fused ring, wherein the number of ring atoms in the aromatic ring includes, but is not limited to, 6 to 18, 6 to 12, or 6 to 10 carbon atoms. The aryl ring can be fused to a saturated or unsaturated carbon ring, wherein the ring connected to the parent structure is the aryl ring. Non-limiting embodiments include benzene rings, naphthalene rings, etc. The "aryl" or "aryl ring" can be monovalent, divalent, trivalent, or tetravalent. When it is divalent, trivalent, or tetravalent, the linking site is located on the aryl ring.

[0160] "Heteroaryl" or "heteroary ring" refers to a substituted or unsubstituted aromatic hydrocarbon group containing 1 to 5 heteroatoms or a group containing heteroatoms (including but not limited to N, O, S(=O)n or Se(=O)n, where n is 0, 1, or 2). The number of ring atoms in the heteroaryl ring includes, but is not limited to, 5 to 15, 5 to 10, or 5 to 6. The atoms C, N, and S on the ring may be optionally oxidized (i.e., C(=O), NO, S(=O)n, Se(=O)n, where n is 1 or 2). Non-limiting examples of heteroaryl groups include, but are not limited to, pyridyl, furanyl, thiophenyl, selenyl, pyridyl, pyranyl, N-alkylpyrrolithyl, pyrimidinyl, pyrazinyl, pyridazinyl, imidazoleyl, benzopyrazolyl, benzimidazoleyl, benzopyridyl, pyrrolopyridyl, pyridinoneyl, etc. The heteroaryl ring can be fused to a saturated or unsaturated carbon ring or heterocycle, wherein the ring connected to the parent structure is an aryl ring. Non-limiting embodiments include: The heteroaryl groups mentioned in this article are defined in accordance with this definition. Heteroaryl groups can be monovalent, divalent, trivalent, or tetravalent. When divalent, trivalent, or tetravalent, the linkage site is located on an aromatic ring.

[0161] "Substituted" or "substituted" means substituted by one or more (including but not limited to 2, 3, 4, or 5) substituents, including but not limited to H, F, Cl, Br, I, alkyl, cycloalkyl, alkoxy, haloalkyl, thiol, hydroxyl, nitro, mercapto, amino, cyano, isocyano, aryl, heteroaryl, heterocyclic, bridged cyclic, spirocyclic, fused cyclic, hydroxyalkyl, =O, carbonyl, aldehyde, carboxylic acid, formate, and -(CH2). m -C(=O)-R a -O-(CH2) m -C(=O)-R a -(CH2) m -C(=O)-NR b R c -(CH2) m S(=O) n R a -(CH2) m -Alkenyl-R a OR d Or -(CH2) m -alkynyl-R a (where m and n are 0, 1, or 2), arylthio, thiocarbonyl, silyl, or -NR b R c Groups, wherein R b With R cIndependently selected from H, hydroxyl, amino, carbonyl, alkyl, alkoxy, cycloalkyl, heterocyclic, aryl, heteroaryl, sulfonyl, trifluoromethanesulfonyl, R b With R c It can form five- or six-membered cycloalkyl or heterocyclic groups, R a With R d Each group is independently selected from aryl, heteroaryl, alkyl, alkoxy, cycloalkyl, heterocyclic, carbonyl, ester, bridged cyclic, spirocyclic, or fused cyclic groups.

[0162] "1 to X substituents selected from..." means substituted by 1, 2, 3...X substituents selected from..., where X is any integer between 1 and 10. For example, "1 to 4 R..." k "Replace" refers to being replaced by 1, 2, 3, or 4 Rs. k Substitution. For example, "1 to 5 substituents selected from ..." means that the ring is substituted by 1, 2, 3, 4 or 5 substituents selected from ... . For example, "the heterobridged ring is optionally substituted by 1 to 4 substituents selected from H or F" means that the heterobridged ring is optionally substituted by 1, 2, 3 or 4 substituents selected from H or F.

[0163] The XY-membered rings (where X and Y are integers, and 3 ≤ X < Y, X < Y ≤ 20, selected from any integer between 4 and 20) include rings of the X, X+1, X+2, X+3, X+4…Y-membered elements. These rings include heterocyclic rings, carbocyclic rings, aromatic rings, aryl groups, heteroaryl groups, cycloalkyl groups, heteromonocyclic rings, heterofused rings, heterospirocyclic rings, or heterobridged rings. For example, "4-7-membered heteromonocyclic rings" refers to heteromonocyclic rings of 4, 5, 6, or 7 members, and "5-10-membered heterofused rings" refers to heterofused rings of 5, 6, 7, 8, 9, or 10 members.

[0164] C x-y Carbocyclic rings (including aryl, cycloalkyl, monocyclic, spirocyclic, fused, or bridged carbocyclic rings) include C x C x+1 C x+2 C x+3 C x+4 ….C y The ring of element (x is an integer, and 3 ≤ x < y, where y is any integer between 4 and 20), for example. "C3-6 cycloalkyl" refers to C3, C4, C5, or C6 cycloalkyl.

[0165] When a functional group has one or more connectable sites, any one or more of these sites can be linked to other functional groups via chemical bonds. When the chemical bond connection is non-directional and a hydrogen atom is present at the connectable site, the number of hydrogen atoms at that site decreases accordingly with the number of bonds being formed, resulting in a functional group with a corresponding valence. For example... This indicates that any connectable site on the piperidinyl group can be linked to other groups via a single chemical bond, including at least... These four connection methods, even if an H atom is drawn on -N-, This also includes For example This indicates that the R group on the piperidinyl group can be located on C or N, and at least includes [missing information]. For example, the general formula segment is: When X is selected from CH2 or NH, it means that the R group on the general formula fragment can be located on C or X. When X is selected from CH2, the general formula fragment can be... When X is selected from NH, the general formula fragment can be:

[0166] Unless otherwise specified, use wedge-shaped solid line keys. and wedge-shaped dashed key The absolute configuration of the center of a solid is represented by a straight solid line key. and straight dashed key Represents the relative configuration of the center of a solid.

[0167] When the listed linking groups do not specify their linking direction, the linking direction includes the direction of the reading order from left to right and from right to left. For example, when ALB is selected from -MW-, it includes AMWB and AWMB.

[0168] "Optional" or "optionally" means that the event or environment described below may but does not have to occur, and the description includes the possibility or possibility that the event or environment may or may not occur. For example, "optionally substituted F alkyl" means that the alkyl group may but does not have to be substituted with F, and the description includes the case where the alkyl group is substituted with F and the case where the alkyl group is not substituted with F.

[0169] "Pharmaceutically acceptable salt" or "its pharmaceutically acceptable salt" means that the compound of the present invention retains the bioavailability and properties of a free acid or a free base, and that the free acid is obtained by reacting with a non-toxic inorganic or organic base, and the free base is obtained by reacting with a non-toxic inorganic or organic acid.

[0170] "Pharmaceutical composition" refers to a mixture of one or more compounds described in this invention, or stereoisomers, tautomers, deuterated compounds, solvates, prodrugs, metabolites, pharmaceutically acceptable salts or cocrystals, and other chemical components, wherein "other chemical components" refers to pharmaceutically acceptable carriers, excipients, and / or one or more other therapeutic agents.

[0171] "Carrier" refers to a material that does not cause significant stimulation to an organism and does not eliminate the biological activity and properties of the compound given.

[0172] "Prodrug" refers to a compound of the present invention that can be metabolized in vivo and converted into a biologically active compound. The prodrug of the present invention is prepared by modifying the amino or carboxyl groups in the compound of the present invention. This modification can be performed through conventional procedures or removed in vivo to obtain the parent compound. When the prodrug of the present invention is administered to a mammalian individual, the prodrug is cleaved to form free amino or carboxyl groups.

[0173] "Co-crystal" refers to a crystal formed by the bonding of an active pharmaceutical ingredient (API) and a co-crystal form (CCF) through hydrogen bonds or other non-covalent bonds. Both API and CCF are solids at room temperature in their pure states, and a fixed stoichiometric ratio exists between the components. Co-crystal is a multi-component crystal, encompassing both binary co-crystals formed between two neutral solids and multi-component co-crystals formed between a neutral solid and a salt or solvate.

[0174] "Stereoisomers" refer to isomers that are produced by different spatial arrangements of atoms in a molecule, including cis-trans isomers, enantiomers, diastereomers, and conformational isomers.

[0175] "Tautomers" refer to functional group isomers that are produced by the rapid movement of an atom in two positions within a molecule, such as keto-enol isomers and amide-imine alcohol isomers.

[0176] "Animals" refers to mammals, such as humans, companion animals, zoo animals, and livestock, with humans, horses, or dogs being preferred.

[0177] IC 50 "It refers to the concentration of a drug or inhibitor required to inhibit a specified biological process (or a component of that process, such as an enzyme, receptor, or cell) by half." Attached Figure Description

[0178] Figure 1 shows the X-ray powder diffraction pattern of the diglucamine salt of compound 2.

[0179] Figure 2 shows the thermogravimetric analysis spectrum of the diglucamine salt of compound 2.

[0180] Figure 3 shows the area under the time-MPT curve of the diglucamine salt of compound 2. Detailed Implementation

[0181] The following embodiments illustrate the technical solution of the present invention in detail, but the scope of protection of the present invention includes, but is not limited to, these embodiments.

[0182] The structure of the compound was determined by nuclear magnetic resonance (NMR) and / or mass spectrometry (MS). NMR shifts (δ) were expressed in 10⁻¹⁰ increments.-6 The unit (ppm) is given. NMR measurements were performed using a Bruker Avance III 400 and Bruker Avance 300 NMR spectrometer. The solvents used were deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), and deuterated methanol (CD3OD). The internal standard was tetramethylsilane (TMS).

[0183] MS determination was performed using (Agilent 6120B (ESI) and Agilent 6120B (APCI));

[0184] HPLC determinations were performed using an Agilent 1260DAD high-performance liquid chromatograph (Zorbax SB-C18 100×4.6mm, 3.5μM).

[0185] Thin-layer chromatography silica gel plates are Yantai Huanghai HSGF254 or Qingdao GF254. The silica gel plates used in thin-layer chromatography (TLC) are 0.15mm-0.20mm in diameter, and the silica gel plates used for thin-layer chromatography separation and purification are 0.4mm-0.5mm in diameter.

[0186] Column chromatography typically uses Yantai Huanghai silica gel 200-300 mesh silica gel as the carrier;

[0187] To accomplish the objectives of this invention, compounds used in the reactions described herein are prepared from commercially available chemicals and / or compounds described in chemical literature, according to organic synthesis techniques known to those skilled in the art. "Commercially available chemicals" are obtained from standard commercial sources, including Shanghai Aladdin Biochemical Technology Co., Ltd., Shanghai Maclean Biochemical Technology Co., Ltd., Sigma-Aldrich, Alfa Esa (China) Chemical Co., Ltd., THIAI (Shanghai) Chemical Industry Development Co., Ltd., Energie Chemicals, Shanghai Titan Technology Co., Ltd., Kelon Chemical, Bailingwei Technology Co., Ltd., etc.

[0188] DIPEA: N,N-Diisopropylethylamine; CAS: 7087-68-5

[0189] HATU: 2-(7-Azobenzotriazole)-N,N,N',N'-Tetramethylurea hexafluorophosphate; CAS: 148893-10-1

[0190] Synthesis Method 1:

[0191] General formulas (Z1) and (Z2) undergo condensation under the conditions of condensing agents such as HATU, and then the protecting groups are removed to obtain the corresponding general formula (I).

[0192] Preparation of intermediate 1a:

[0193] 298 mg of A (synthesis method referred to Example 43 of WO2024146632) was prepared by SFC, and after lyophilization, white solids B (chiral HPLC retention time: 1.079 min) and 1a (98 mg, chiral HPLC retention time: 1.273 min) were obtained.

[0194] Chiral HPLC: (Instrument: SHIMADZU LC-30AD sf, Chiral column: Chiral IG column. Preparation method: The crude product was dissolved in acetonitrile to prepare a sample solution. Mobile phase system: carbon dioxide / 0.05% DEA in ethanol solution. Elution gradient: 5%-40%; flow rate: 3.0 mL / min, elution time: 3 min).

[0195] SFC preparation conditions:

[0196] Instrumentation: Waters 150Prep-SFC; Preparative column: Chiral IG column. Preparation method: The crude product was dissolved in acetonitrile and methanol to prepare a 10 mg / ml sample solution. Mobile phase: Carbon dioxide / ethanol, ethanol content 20%; Elution time: 3.5 min.

[0197] Intermediate 1a: LCMS m / z = 541.2 [M+H] + ;

[0198] 1 H NMR(400MHz,DMSO-d6)δ10.70(s,1H),8.13–8.07(m,1H),7.81–7.75(m,1H),7.36(t,1H),7.23–7.14(m,1H),7.12–7.04(m,1 H),4.85(d,1H),4.64(s,1H),4.53–4.45(s,1H),4.1(d,3H),3.15(s,3H),2.67–2.58(m,1H),1.86(s,3H),0.84–0.74(m,3H).

[0199] Example 1: Preparation of Compound 1

[0200] Step 1: Preparation of 1b

[0201] 1a (108 mg, 0.20 mmol) was dissolved in ultra-dry DMF (2 mL), and (S)-2,6-di-tert-butoxycarbonylaminohexanoic acid (104 mg, 0.30 mmol), HATU (152 mg, 0.40 mmol), and DIPEA (0.1 mL, 0.61 mmol) were added sequentially under ice bath conditions. The mixture was then stirred at room temperature for 16 hours. 5 mL of water was added to the reaction mixture, and the mixture was extracted with ethyl acetate (5 mL × 3). The mixture was washed with saturated sodium chloride aqueous solution (5 mL × 3), and the organic layers were combined and dried over anhydrous sodium sulfate. The mixture was filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography (PE / EA = 2 / 1 (v / v)) to obtain 1b (160 mg, yield 92.16%).

[0202] Step 2: Preparation of Compound 1

[0203] Compound 1 (160 mg, 0.18 mmol) was dissolved in dichloromethane (2 mL), and 4 M hydrochloric acid-dioxane solution (2 mL) was added. The mixture was reacted at room temperature for 1.5 hours. The mixture was concentrated to dryness and purified by preparative liquid chromatography (instrument: Waters 2767 preparative liquid chromatography; column: SunFire@Prep C18 (19 mm × 250 mm); mobile phase composition: mobile phase A: acetonitrile / mobile phase B: water (containing 0.05% HCl) to obtain compound 1 (84 mg, yield 61.51%).

[0204] LCMS m / z = 669.2 [M+H] +

[0205] 1 H NMR(400MHz,DMSO-d6)δ11.53-11.38(brs,1H),8.41-8.34(m,1H),8.28-8.12(m, 3H),8.07-7.83(m,4H),7.55-7.45(m,1H),7.36-7.08(m,2H),5.16-5.07(m,1H),4 .54-4.45(m,1H),4.03-3.96(m,3H),3.82-3.72(m,1H),3.61(s,3H),2.83-2.68(m ,2H),2.65-2.56(m,1H),1.93-1.73(m,5H),1.65-1.31(m,4H),0.85-0.70(m,3H).

[0206] Compound 1 was dissolved in ethyl acetate, and various alkaline reagents such as sodium bicarbonate, potassium carbonate, sodium hydroxide, triethylamine, and diisopropylethylamine were tried, but the pure free form of compound 1 could not be obtained.

[0207] Example 2: Preparation of Compound 2

[0208] Step 1: Preparation of Compound 2

[0209] 1a (108 mg, 0.20 mmol) was dissolved in ultra-dry dichloromethane (2 mL), and triethylamine (0.35 mL, 2.52 mmol) was added. Phosphorus oxychloride (0.2 mL, 2.18 mmol) was added dropwise under ice bath conditions. The reaction was carried out at room temperature for 16 hours. 5 mL of ice water was added to the reaction solution and stirred for 1 hour. The mixture was extracted with ethyl acetate (5 mL × 3), washed with saturated sodium chloride aqueous solution (5 mL), and the organic layers were combined and dried over anhydrous sodium sulfate. The mixture was filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by preparative liquid chromatography (instrument: Waters 2767 preparative liquid chromatography; column: SunFire@Prep C18 (19 mm × 250 mm); mobile phase composition: mobile phase A: acetonitrile / mobile phase B: water (containing 0.1% TFA) to obtain compound 2 (30 mg, yield 24.20%).

[0210] LCMS m / z = 621.2[M+H] +

[0211] 1 H NMR(400MHz,DMSO-d6)δ11.05-10.80(brs,1H),8.14-8.06(m,1H),7.97-7.86(m,1H),7.42-7.34(m,1H),7.21-6.97(m,4H) ,4.92-4.81(m,1H),4.54-4.43(m,1H),4.03-3.94(m,3H),3.38(s,3H),2.63-2.56(m,1H),1.85(s,3H),0.82-0.69(m,3H).

[0212] Example 3: Preparation of the diglucamine salt of compound 2

[0213] Step 1: Preparation of diglucamine salt of compound 2

[0214] Compound 2 (500 mg, 0.81 mmol) was dissolved in methanol (10 mL), and meglumine (320 mg, 1.64 mmol) was added. After the system clarified, it was stirred for 30 minutes, concentrated to dryness, and isopropanol (15 mL) was added. After stirring for 1 h, a solid precipitated. The solid was collected by filtration, dried under reduced pressure, and dissolved in water (10 mL). After clarification, it was lyophilized to obtain the diglumine salt of compound 2 (700 mg, yield 85.93%). The amorphous form of the diglumine salt of compound 2 was characterized by XRPD and TGA, as shown in Figures 1 and 2, respectively.

[0215] 1 H NMR(400MHz, DMSO-d6)δ11.66-11.33(brs,1H),8.19-7.96(m,2H),7.37-7.00(m,3H),5.05-4.98(m,1H),4.51-4.43(m,1H),4.01-3.94(m,3H) ,3.84-3.76(m,2H),3.68-3.55(m,4H),3.52-3.30(m,9H),2.84-2.67( m,4H),2.59-2.53(m,1H),2.38(s,6H),1.84(s,3H),0.82-0.63(m,3H).

[0216] Example 4: Preparation of Compound 4

[0217] Step 1: Preparation of 4b

[0218] 1a (100 mg, 0.19 mmol) was mixed with N-Boc-L-alanine (43.14 mg, 0.23 mmol) according to the method in step one of Example 1 to obtain 4b (71 mg, yield 52.43%).

[0219] Step 2: Preparation of Compound 1

[0220] Compound 4 (32 mg, yield 52.52%) was obtained by referring to the method in step 2 of Example 1, with 4b (71 mg, 0.10 mmol).

[0221] LCMS m / z = 612.2[M+H] +

[0222] 1 H NMR (400MHz, DMSO-d6) δ11.26-11.13(m,1H),8.38-8.28(m,1H),8.21-7.64(m,4H),7.54-7.43(m,1H),7.24-7.10(m,2H),5.50-4.87(m ,1H),4.58-4.41(m,1H),4.00(s,3H),3.90-3.78(m,1H),3.59(s,3H),2.65-2.55(m,1H),1.86(s,3H),1.40(d,3H),0.83-0.70(m,3H).

[0223] Example 5: Preparation of Compound 5

[0224] Step 1: Preparation of 5b

[0225] 1a (100 mg, 0.19 mmol) was mixed with N-Boc-L-cyclopropylglycine (49.08 mg, 0.23 mmol) according to the method in step one of Example 1 to obtain 5b (75 mg, yield 54.88%).

[0226] Step 2: Preparation of Compound 1

[0227] Compound 5 (24 mg, yield 37.07%) was obtained by referring to the method in step 2 of Example 1 with 5b (75 mg, 0.10 mmol).

[0228] LCMS m / z = 638.2 [M+H] +

[0229] 1 H NMR(400MHz,DMSO-d6)δ11.08(s,1H),8.40-8.31(m,1H),8.13(s,3H),7. 90-7.80(m,1H),7.55-7.44(m,1H),7.22-7.07(m,2H),5.01-4.85(m,1H), 4.55-4.43(m,1H),4.00(s,3H),3.60(s,3H),3.18(d,1H),2.66-2.57(m,1 H),1.85(s,3H),1.12-0.95(m,1H),0.83-0.72(m,3H),0.68-0.46(m,4H).

[0230] Example 6: Preparation of Compound 6

[0231] Step 1: Preparation of 6a

[0232] 1a (540 mg, 1.0 mmol) was dissolved in DCM (5 mL), and chloromethyl chloroformate (1.0 g, 7.76 mmol) and DIPEA (1.7 mL, 10.29 mmol) were added sequentially under ice bath conditions. The mixture was stirred at room temperature for 16 hours. 5 mL of water was added to the reaction solution, and the mixture was extracted with ethyl acetate (5 mL × 3). The mixture was washed with saturated sodium chloride aqueous solution (5 mL × 3), and the organic layers were combined and dried over anhydrous sodium sulfate. The mixture was filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography (PE / EA = 1 / 1 (v / v)) to obtain 6a (430 mg, yield 68.00%).

[0233] Step 2: Preparation of compound 6b

[0234] 6a (430 mg, 0.68 mmol) was dissolved in THF (30 mL), and tetrabutylammonium di-tert-butyl phosphate (920 mg, 2.04 mmol) and sodium iodide (150 mg, 1.00 mmol) were added sequentially. The mixture was heated to 70°C and stirred for 1 hour. 20 mL of water was added to the reaction mixture, and the mixture was extracted with ethyl acetate (20 mL × 3). The mixture was washed with saturated sodium chloride aqueous solution (20 mL × 1), and the organic layers were combined and dried over anhydrous sodium sulfate. The mixture was filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography (PE / EA = 1 / 1 (v / v)) to obtain 6b (490 mg, yield 89.41%).

[0235] Step 3: Preparation of Compound 6

[0236] 6b (200 mg, 0.25 mmol) was dissolved in dichloromethane (10 mL), and trifluoroacetic acid (2 mL) was added. The mixture was reacted at room temperature for 1 hour, and the system was concentrated to dryness. The solution was then purified by preparative liquid chromatography (instrument: Waters 2767 preparative liquid chromatography; column: SunFire@Prep C18 (19 mm × 250 mm); mobile phase composition: mobile phase A: acetonitrile / mobile phase B: water (containing 0.1% TFA) to obtain compound 6 (60 mg, yield 34.85%).

[0237] LCMS m / z = 693.2 [MH] +

[0238] 1 H NMR(400MHz,DMSO-d6)δ10.84(s,1H),8.25-8.19(m,1H),7.92-7.84(m,1H),7.52-7.44(m,1H),7.25-7.14(m,1H),7.13-7.05(m,1H),5.4 1-5.24(m,2H),4.90-4.78(m,1H),4.55-4.43(m,1H),4.04-3.96(m,3H),3.55(s,3H),2.66-2.58(m,1H),1.86(s,3H),0.87-0.72(m,3H).

[0239] Example 7: Preparation of Compound 7

[0240] Step 1: Preparation of 7a

[0241] Compound 7a (200 mg, 96.84%) was obtained by referring to the first step of Example 1, using 1a (160 mg, 0.30 mmol) and Boc-glycine (70 mg, 0.40 mmol) as starting materials.

[0242] Step 2: Preparation of Compound 7

[0243] Compound 7 (50 mg, yield 27.51%) was obtained from 7a (200 mg, 0.29 mmol) using the method in step 2 of Example 1.

[0244] LCMS m / z = 598.3 [M+H] +

[0245] 1 H NMR (400MHz, DMSO-d6) δ11.28-11.03(brs,1H),8.39-8.28(m,1H),8.14-7.69(m,4H),7.54-7.43(m,1H),7.24-7.11(m,2H),5.04-4. 87(m,1H),4.56-4.43(m,1H),4.03-3.97(m,3H),3.71-3.60(m,2H),3.57(s,3H),2.67-2.55(m,1H),1.86(s,3H),0.85-0.69(m,3H).

[0246] Example 8: Preparation of Compound 8

[0247] Step 1: Preparation of 8a

[0248] Compound 8a (150 mg, 0.28 mmol) was obtained from 1a (150 mg, 0.28 mmol) and Boc-D-alanine (79 mg, 0.42 mmol) using the method of step one of Example 1.

[0249] Step 2: Preparation of Compound 8

[0250] Compound 8 (80 mg, yield 58.57%) was obtained from 8a (150 mg, 0.21 mmol) using the method in step 2 of Example 1.

[0251] LCMS m / z = 612.3[M+H] +

[0252] 1H NMR(400MHz,DMSO-d6)δ11.24-10.91(brs,1H),8.40-8.30(m,1H),8.12-7.79(m,4H),7.55-7.43(m,1H),7.24-7.09(m,2H),5.03-4.86(m,1H) ,4.55-4.44(m,1H),4.04-3.95(m,3H),3.92-3.82(m,1H),3.57(s,3H) ,2.65-2.57(m,1H),1.86(s,3H),1.38-1.29(m,3H),0.83-0.74(m,3H).

[0253] Example 9: Preparation of Compound 9

[0254] Step 1: Preparation of 9a

[0255] Compound 9a (120 mg, 56.03%) was obtained from 1a (150 mg, 0.28 mmol) and Boc-L-methionine (138 mg, 0.55 mmol) using the method of step one of Example 1.

[0256] Step 2: Preparation of Compound 9

[0257] Compound 9 (70 mg, yield 63.58%) was obtained from 9a (120 mg, 0.16 mmol) using the method in step 2 of Example 1.

[0258] LCMS m / z = 672.3 [M+H] +

[0259] 1 H NMR(400MHz,DMSO-d6)δ11.17-10.98(brs,1H),8.39-8.31(m,1H),8.27-7.76(m,4H),7.55-7.45(m,1H),7.23-7.10(m,2H),5.00-4.89(m,1H) ,4.53-4.45(m,1H),4.02-3.98(m,3H),3.94-3.87(m,1H),3.62(s,3H) ,2.66-2.53(m,3H),2.17-1.94(m,5H),1.86(s,3H),0.86-0.71(m,3H).

[0260] Example 10: Preparation of Compound 10

[0261] Step 1: Preparation of 10a

[0262] Compound 10a (140 mg, 68.20%) was obtained using 1a (150 mg, 0.28 mmol) and Boc-L-valine (120 mg, 0.55 mmol) as starting materials, following the method in step one of Example 1.

[0263] Step 2: Preparation of Compound 10

[0264] Compound 10 (60 mg, yield 46.89%) was obtained from 10a (140 mg, 0.19 mmol) using the method in step 2 of Example 1.

[0265] LCMS m / z = 640.4 [M+H] +

[0266] 1 H NMR(400MHz,DMSO-d6)δ11.29-11.03(m,1H),8.41-8.31(m,1H),8.11-7.75 (m,4H),7.56-7.44(m,1H),7.24-7.09(m,2H),5.04-4.90(m,1H),4.55-4.4 5(m,1H),4.02-3.97(m,3H),3.72-3.66(m,1H),3.61(s,3H),2.65-2.56(m, 1H),2.31-2.18(m,1H),1.86(s,3H),1.02-0.90(m,6H),0.83-0.72(m,3H).

[0267] Example 11: Preparation of Compound 11

[0268] Step 1: Preparation of Compound 11

[0269] Dissolve 1a (1.08 g, 2.0 mmol) in dichloromethane (10 mL), and add triethylamine (607.14 mg, 6.0 mmol) sequentially under ice bath conditions.

[0270] 4-Dimethylaminopyridine (24.43 mg, 0.2 mmol) and methanesulfonyl chloride (458.2 mg, 4.0 mmol) were reacted at room temperature for 18 hours. The reaction was quenched with water (15 mL), extracted with dichloromethane (15 mL × 3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered and concentrated to obtain the crude product, which was purified by silica gel column chromatography to give compound 11 (280.0 mg, yield 22.63%).

[0271] LCMS m / z = 619.1 [M+H] +

[0272] 1 H NMR (400MHz, DMSO-d6) δ10.84(m,1H),8.27-8.21(m,1H),7.90-7.82(m,1H),7.51(t,1H),7.24-7.13(m,1H),7.11-7.04(m,1H ),4.84(d,1H),4.55-4.44(m,1H),4.00(d,3H),3.61(s,3H),2.98(s,3H),2.68-2.57(m,1H),1.86(s,3H),0.86-0.72(m,3H).

[0273] Example 12: Preparation of Compound 12

[0274] Step 1: Preparation of compound 12b

[0275] 12a (1.0 g, 2.82 mmol) was dissolved in THF (16 mL) and water (4 mL), and 12a-1 (0.89 g, 3.10 mmol) and N-methylmorpholine (0.37 mL, 3.38 mmol) were added. The mixture was reacted at room temperature for 16 hours. 50 mL of water was added to the reaction solution, and the solid was filtered and dissolved in dichloromethane (50 mL). The solid was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give 12b (1.66 g, yield 94.16%).

[0276] LCMS m / z = 525.2 [M-99] +

[0277] Step 2: Preparation of compound 12c

[0278] 12b (1.66 g, 2.66 mmol) was dissolved in ethanol (20 mL), and hydrochloric acid (1.21 mL, 39.9 mmol) was added. The mixture was reacted at 55 °C for 4 hours. The solvent was removed, and the mixture was dissolved in ethyl acetate (50 mL) and the pH was adjusted to 9 with NHCO3 aqueous solution. The mixture was washed with saturated sodium chloride aqueous solution (50 mL), and the organic layers were combined and dried over anhydrous sodium sulfate. The mixture was filtered, and the filtrate was concentrated under reduced pressure to give compound 12c (1.3 g, yield 93.26%).

[0279] LCMS m / z = 525.2 [M+H] +

[0280] Step 3: Preparation of Compound 12

[0281] 12c (1.3 g, 2.48 mmol) was dissolved in ultra-dry dichloromethane (20 mL), and triethylamine (0.69 mL, 4.96 mmol) was added. Phosphorus oxychloride (0.34 mL, 3.72 mmol) was added dropwise under ice bath conditions. The reaction was carried out at room temperature for 16 hours. 50 mL of ice water was added to the reaction solution and stirred for 1 hour. The mixture was extracted with ethyl acetate (50 mL × 3), washed with saturated sodium chloride aqueous solution (50 mL), and the organic layers were combined and dried over anhydrous sodium sulfate. The mixture was filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by preparative liquid chromatography (instrument: Waters 2767 preparative liquid chromatography; column: SunFire@Prep C18 (19 mm × 250 mm); mobile phase composition: mobile phase A: acetonitrile / mobile phase B: water (containing 0.1% TFA) to obtain compound 12 (600 mg, yield 40.04%).

[0282] LCMS m / z = 605.2[M+H] +

[0283] 1 H NMR(400MHz,DMSO-d6)δ11.61(s,1H),8.25-8.17(m,1H),8.02-7.91(m,1H),7.43(t,1H),7.23-7.09(m,2H), 5.07(d,1H),4.29-4.20(m,1H),3.95(s,3H),3.40(s,3H),2.84-2.70(m,1H),1.60(s,3H),0.78-0.67(m,3H).

[0284] Biological test example 1

[0285] Nav1.8 Manual Patch Clamp Test

[0286] (1) Cell Culture

[0287] The CHO cell line stably expressing human Nav1.8 was cultured in Ham's F-12 medium containing 10% fetal bovine serum and 10 μg / mL Blasticidin, 200 μg / mL Hygromycin B, and 100 μg / mL Zeocin. The cell culture temperature was 37°C and the carbon dioxide concentration was 5%. After removing the old medium and washing once with PBS, 1 mL of 0.25% Trypsin-EDTA solution was added, and the cells were incubated at 37°C for approximately 1.5 min. When the cells detached from the bottom of the dish, pre-warmed complete medium (37°C) was added. The cell suspension was gently pipetted to separate aggregated cells. The cell suspension was transferred to sterile centrifuge tubes and centrifuged at 1000 rpm for 5 min to collect the cells. The cells were seeded in 6 cm cell culture dishes at a density of 2.5 × 10⁶ cells per dish. 5 Cells (final volume 5 mL) were used for expansion or maintenance culture. To maintain cell electrophysiological activity, the cell density should not exceed 80%. Before patch-clamp detection, cells were separated with 0.25% Trypsin-EDTA, and 6.5 × 10⁶ cells were cultured. 3 Cells were seeded onto coverslips and cultured in 24-well plates (final volume 500 μL), and analyzed after 18 hours.

[0288] (2) Compound preparation

[0289] The compound was dissolved in dimethyl sulfoxide (DMSO) to prepare a 30 mM DMSO stock solution. The stock solution was diluted to the test concentration with extracellular fluid (140 mM NaCl, 3.5 mM KCl, 1 mM MgCl2·6H2O, 2 mM CaCl2·2H2O, 10 mM D-Glucose, 10 mM HEPES and 1.25 mM NaH2PO4·2H2O, pH adjusted to 7.4 with NaOH). The final DMSO concentration of all test samples was 0.1%.

[0290] (3) Electrophysiological tests

[0291] First, a capillary glass tube was drawn into a recording electrode using a microelectrode drawing device. Then, the electrode, filled with intracellular fluid (50 mM CsCl, 10 mM NaCl, 10 mM HEPES, 60 mM CsF, and 20 mM EGTA, pH adjusted to 7.2 with CsOH), was placed into a microelectrode holder. Under an inverted microscope, the microelectrode manipulator was used to immerse the electrode in the extracellular fluid, and the electrode resistance (Rpip) was recorded. Next, the electrode was slowly brought into contact with the cell surface, and negative pressure was applied to form a GΩ seal. Fast capacitance compensation was then performed, and negative pressure was continued to rupture the cell membrane, establishing a whole-cell recording mode. Finally, slow capacitance compensation was performed, and experimental parameters such as series resistance (Rs) were recorded. No leakage compensation was applied. Once the Nav1.8 current recorded in the whole cell stabilized, drug administration began, with each drug concentration acting for approximately 5 minutes (or until the current stabilized). A coverslip containing cells was placed in a recording bath under an inverted microscope. Blank control solution and the working solution of the test compound were perfused through the recording bath by gravity to act on the cells, with fluid exchange facilitated by a peristaltic pump. The current detected in the cells in the solution without the compound served as a control group. All electrophysiological experiments were performed at room temperature. The inhibitory rate of the compound on Nav1.8 was determined by calculating the relative percentage of peak currents generated before and after cell treatment.

[0292] The voltage stimulation protocol for whole-cell patch-clamp recording of Nav1.8 sodium current is as follows: After whole-cell sealing, the cell voltage is clamped at -120 mV. The voltage is first stepped from -110 mV to -30 mV in 10 mV increments, maintained for 5 seconds, and then a 0 mV depolarization pulse is applied to obtain the half-inactivation voltage (V). half Then with V half The stimulation voltage was maintained for 5 seconds, then restored to -120 mV and maintained for 20 ms. A depolarization pulse (TP2) was then applied to 0 mV for 50 ms to detect the sodium current in the semi-inactivated state. Finally, the voltage was restored to the clamp voltage of -120 mV, and data was collected every 20 ms to observe the effect of the drug on the peak sodium current. Experimental data were acquired using an EPC 10 amplifier (HEKA) and stored in PatchMaster (HEKA) software.

[0293] Table 1 shows the IC50 values ​​of the tested compounds for inhibiting Nav1.8. 50

[0294] Conclusion: The compounds of the present invention, such as the compounds in the examples, have good Nav1.8 inhibitory activity.

[0295] Nav1.4 and Nav1.6 manual patch clamp tests

[0296] CHO or HEK-293 cell lines expressing human Nav1.4 and Nav1.6 channels, respectively, were constructed in the laboratory of Beijing Aisiyipu Biotechnology Co., Ltd. The gene information is as follows: Nav1.4 (NM_000334), Nav1.6 (NM_014191).

[0297] Nav1.4 test

[0298] Culture preparation: CHO cells were cultured in Ham's F-12 medium containing 10% fetal bovine serum at a cell culture incubator temperature of 37°C and a carbon dioxide concentration of 5%.

[0299] Cell passage: Remove the culture medium and gently rinse the cell surface with PBS. Then, add 1 mL of 0.25% Trypsin-EDTA solution to digest the cells and incubate the culture dish at 37°C for approximately 1.5 min. When the cells detach from the bottom of the dish, add approximately 5 mL of preheated 37°C complete culture medium. Gently pipette the cell suspension to disperse any aggregated cell clusters, then transfer to a sterile centrifuge tube and collect the cells by centrifugation at 1000 rpm for 5 min. In standard culture, seed the cells at a density of 2.5 × 10⁵ cells per 6 cm culture dish (final volume: 5 mL).

[0300] To maintain the electrophysiological activity of cells, cell confluence should not exceed 80%.

[0301] Before patch-clamp assays, cells were separated with 0.25% Trypsin-EDTA and seeded at a density of 8 × 10³ cells per well into 24-well plates with cell spreaders (final volume: 500 μL). After induction with tetracycline for 24–72 hours, electrophysiological experiments were performed.

[0302] Nav1.6 test

[0303] Culture preparation: HEK-293 cells were cultured in DMEM medium containing 10% fetal bovine serum and 800 μg / mL G418, at a cell culture incubator temperature of 37°C and a carbon dioxide concentration of 5%.

[0304] Cell passage: Remove the culture medium and gently rinse the cell surface with PBS. Then, add 1 mL of 0.25% Trypsin-EDTA solution to digest the cells and incubate the culture dish at 37°C for about 1 min. When the cells detach from the bottom of the dish, add about 5 mL of preheated 37°C complete culture medium. Gently pipette the cell suspension to disperse any aggregated cell clumps, then transfer to a sterile centrifuge tube and centrifuge at 1000 rpm for 5 min to collect the cells. In standard culture, seed the cells at a density of 2.5 × 10⁵ cells per 6 cm culture dish (final volume: 5 mL).

[0305] To maintain the electrophysiological activity of cells, cell confluence should not exceed 80%.

[0306] Before patch-clamp assays, cells were separated with 0.25% Trypsin-EDTA and seeded at a density of 8 × 10³ cells per well into 24-well plates with cell spreaders (final volume: 500 μL). Electrophysiological experiments were performed 18 hours later.

[0307] Electrophysiological recording

[0308] Record the liquid used

[0309] Extracellular fluid: K-007-1

[0310] 140mM NaCl, 3.5mM KCl, 1mM MgCl₂·6H₂O, 2mM CaCl₂·2H₂O, 10mM D-Glucose, 10mM HEPES, 1.25mM NaH₂PO₄·2H₂O, pH adjusted to 7.4 with NaOH.

[0311] Intracellular fluid: Nav-001-2

[0312] Adjust pH to 7.2 with 50mM CsCl, 10mM NaCl, 10mM HEPES, 60mM CsF, 20mM EGTA, and CsOH.

[0313] Extracellular fluid should be stored at 4°C and used within 2 weeks. Intracellular fluid, after preparation, should be aliquoted into 1 mL tubes and stored at -20°C. Freshly thawed intracellular fluid should be used daily for experiments. All intracellular fluid should be used within three months; otherwise, discard and prepare fresh.

[0314] Patch clamp testing

[0315] The voltage stimulation protocol for whole-cell patch-clamp recording of sodium current was as follows: After whole-cell sealing, the cell voltage was clamped at -120 mV for 5 seconds, followed by a depolarization pulse (TP1) to 0 mV for 20 ms, and finally returned to the clamp voltage of -120 mV. Data acquisition was repeated at 20-second intervals to observe the effect of the drug on the peak sodium current at rest. Experimental data were acquired using an EPC 10 amplifier (HEKA) and stored in PatchMaster (HEKA) software.

[0316] The patch-clamp procedure begins by using a microelectrode puller to draw a glass capillary into a recording electrode. The electrode, filled with intracellular fluid, is then placed into a microelectrode holder. Next, a cell slide coated with cells is placed in a recording bath under an inverted microscope. Under the microscope, the microelectrode manipulator is manipulated to immerse the electrode in the extracellular fluid, and the electrode resistance (Rpip) is recorded. The electrode is then slowly brought into contact with the cell surface, and negative pressure is applied to create a GΩ high-resistance seal. Fast capacitance electrode compensation is then performed, and negative pressure is continued to rupture the cell membrane, establishing a whole-cell recording mode. Finally, slow capacitance cell compensation is performed, and experimental parameters such as series resistance (Rs) are recorded. No leakage compensation is applied.

[0317] Once the current amplitude in the cells stabilized in the solvent control, drug administration began, followed by sequentially increasing the concentration of the working solution of the test compound. Each concentration was applied until the current response reached equilibrium (within approximately 5 minutes), at which point the next concentration was introduced. All solutions were applied to the cell surface via a gravity perfusion system, while a peristaltic pump continuously replaced the fluid in the recording bath. Electrophysiological recording was performed at room temperature.

[0318] Data quality control standards

[0319] The following criteria are used to determine whether data is acceptable:

[0320] Electrode resistance <5MΩ; Sealing resistance >1GΩ

[0321] Initial connection resistance <15MΩ

[0322] The input resistance ends at <15MΩ

[0323] Peak starting current >300pA

[0324] The current did not exhibit significant spontaneous decay.

[0325] At the same concentration, the difference in inhibition rate between repeated data is ≤15%.

[0326] Data Analysis

[0327] Peak sodium current after each drug concentration was applied. compound ) and peak current (contrast current) control Normalize, then calculate the inhibition rate corresponding to each drug concentration, i.e., Inhibition% = (1 - (Peak current)) / (Peak current) compound ) / (Peak current control ))*100%, and calculate the mean (Mean), standard deviation (SD), and standard error (SE) for each drug concentration inhibition rate. The data are expressed as Mean±SE.

[0328] A four-parameter nonlinear fitting was performed on the concentration-inhibition rate curve. Absolute IC50 and IC90 were calculated through interpolation, specifically obtaining the X values ​​corresponding to Y = 50 and Y = 90, respectively. Here, IC50 is the half-inhibitory concentration, IC90 is the concentration at which the inhibition rate is 90%, X is the logarithm of the concentration, and Y is the inhibition rate. The calculation of IC50 and IC90, as well as curve fitting, were performed using GraphPad Prism software.

[0329] Conclusion: The compounds of the present invention have good family selectivity. For example, the diglucamine salt of compound 2 has an IC50 greater than 10 μM for both Nav1.4 and Nav1.6 channels, and has no obvious inhibitory effect.

[0330] Biological Test Example 2: Spinal Nerve Ligation (SNL) Induced Mouse Model of Neuropathic Pain

[0331] Male C57BL / 6J mice (7-8 weeks old) purchased from Zhejiang Vital River Laboratory Animal Technology Co., Ltd. were acclimatized for one week before the model was established. The specific establishment method is as follows:

[0332] (1) Disinfection of surgical instruments and ligation sutures;

[0333] (2) Anesthetize the mice with isoflurane and place them in a prone position on the operating table;

[0334] (3) Clip the hair and prepare the skin near the hip bone of the mouse, and make an incision of about 2 cm along the spine;

[0335] (4) Dissect the fascia along the spine, bluntly dissect the muscles, and expose the L5 transverse process;

[0336] (5) Carefully bite off the L5 transverse process with tweezers to expose the L5 spinal nerve;

[0337] (6) Carefully separate the L5 nerve with a glass needle and ligate the L5 nerve with 5-0 ligation suture;

[0338] (7) Suture the muscles and skin, and disinfect with povidone-iodine.

[0339] Mice that failed to establish the model were culled the day after model establishment (successful model indicator: hind paw curled). After model establishment, mice were petted for 3-5 minutes daily to ensure familiarity with the experimenters, followed by placing them on a metal pain assessment frame for 40-60 minutes to acclimatize. After environmental acclimatization on day 3, Von Frey fibers (…) were used… The baseline values ​​of mice were measured before administration (using the Ascending test method) at doses of 0.16, 0.4, 0.6, 1.0, 1.4, and 2.0 g. Each animal was measured twice, and the average value was taken, with an interval of at least 5 minutes between each measurement. Animals were grouped according to their baseline values ​​(10 animals per group). On day 4 post-surgery, compound 2 was administered via tail vein injection in either diglucamine salt or a solvent (physiological saline, model group). The amount of compound 2 was calculated when determining the dosage. The mechanical pain threshold (MPT) of mice was measured at 2, 4, 6, and 8 hours post-administration, and the area under the time-MPT curve (AUC) was calculated. MPT Statistical analysis was performed using GraphPad 8.3.0. The results are shown in Figure 3.

[0340] Results and conclusions: At doses of 10 and 30 mg / kg, compound 2 significantly increased the mechanical pain threshold in mice, indicating that compound 2 has good analgesic efficacy.

[0341] Biological Test Example 3: CYP450 Enzyme Inhibition Test

[0342] The aim of this study was to evaluate the effects of test substances on the activities of five isoenzymes (CYP1A2, CYP2C9, CYP2C19, CYP2D6, and CYP3A4) of human liver microsomal cytochrome P450 (CYP) using an in vitro assay system. Specific probe substrates for CYP450 isoenzymes were co-incubated with human liver microsomes and different concentrations of the test substances. Reduced nicotinamide adenine dinucleotide phosphate (NADPH) was added to initiate the reaction. After the reaction, the metabolites produced by the specific substrates were quantitatively detected by liquid chromatography-tandem mass spectrometry (LC-MS / MS) after sample processing, and the changes in CYP enzyme activity were measured. The IC50 values ​​were calculated. 50 The value is used to evaluate the inhibitory potential of the test substance against each CYP enzyme subtype.

[0343] Conclusion: The compounds of the present invention, such as the compounds in the examples, have weak CYP inhibition. For example, compound 2 has an IC50 greater than 30 μM against the five isoenzymes of CYP (CYP1A2, CYP2C9, CYP2C19, CYP2D6 and CYP3A4).

[0344] Biological Test Example 4: Single Intravenous Injection Tolerance Test in SD Rats

[0345] Female SD rats, 6-8 weeks old and weighing 205-237g, were purchased from Sichuan Vital River Laboratory Animal Technology Co., Ltd. The specific experimental methods are as follows:

[0346] (1) SD rats were randomly divided into 6 groups, namely compound 1: 2.5, 10, 50 mg / kg dose group and compound 2: 2.5, 10, 50 mg / kg dose group, with 3 rats in each group;

[0347] (2) Prepare compound 1 and compound 2 at the corresponding concentrations using physiological saline; if the dissolution is incomplete, adjust the pH to 4-9 until the dissolution is complete.

[0348] (3) Each group of rats was intravenously injected with the corresponding dose of compound 1 and compound 2 at a volume of 5 mL / kg according to the group.

[0349] (4) Closely observe after medication;

[0350] Results: No significant abnormalities were observed in rats after administration of compound 2 at any dosage group. Compound 2 exhibits good safety. One rat in the 12.5 mg / kg group died after administration, and all rats in the 10 mg / kg and 50 mg / kg dosage groups died within 1 to 5 minutes after administration.

[0351] Biological Test Example 5: UGT1A1 Inhibitory Activity Test

[0352] This experiment evaluated the inhibitory potential of test substances on UGT1A1 using recombinant human UGT1A1 enzyme. Bilirubin, a probe substrate for the UGT1A1 enzyme, was co-incubated with recombinant human UGT1A1 enzyme and different concentrations (0–10 μM) of the test substances. Uridine diphosphate glucuronide (UDPGA) was added to initiate the reaction. After the reaction, the samples were processed, and specific metabolites produced by bilirubin were quantitatively detected using liquid chromatography-tandem mass spectrometry (LC-MS / MS). Changes in UGT1A1 enzyme activity were measured, and the IC50 was calculated. 50 The value is used to evaluate the inhibitory potential of the test substance on each UGT1A1 enzyme.

[0353] Conclusion: The compounds of the present invention, such as the diglucamine salt of compound 2, showed an inhibition rate of less than 50% against UGT1A1 at 10 μM, suggesting that compound 2 has a low risk of abnormal bilirubin-related indicators caused by UGT1A1 at a concentration of 10 μM.

[0354] Biological Test Example 6: CYP3A4 Induction Activity Assay

[0355] 1. Cell inoculation

[0356] 1) DPX2 cells were cultured in growth medium containing 10% fetal bovine serum.

[0357] 2) DPX2 cells were cultured in T-75 culture flasks in an incubator at 37°C, 5% CO2 and 95% relative humidity. The cells were digested when they reached 80-90% confluence with the bottom of the culture flask.

[0358] 3) Wash the surface of T-75 cultured cells with 10mL PBS, remove the PBS, add 3-5mL trypsin, digest at 37℃ for 5 minutes or until the cells are digested and suspended, and add excess culture medium containing fetal bovine serum to stop the trypsin digestion.

[0359] 4) Transfer the cell suspension to a conical-bottom centrifuge tube and centrifuge at 150g for 5 minutes at room temperature. Carefully aspirate the supernatant, resuspend the cells in treatment culture medium, and adjust the concentration to 3.2 × 10⁻⁶. 5 cells / mL (incubation time 24 hours, seed plate density 4.0 × 10⁻⁶) 5 Add 25 μL of cell suspension to each well of a 384-well cell culture plate. Incubate the cell culture plate at 37°C, 5% CO2, and 95% relative humidity for 24 hours.

[0360] 2. Compound preparation

[0361] 1) Prepare stock solutions of the test compound, positive control (rifampin), and negative control (propranolol) at a concentration of 1000× using DMSO. The final concentrations of the positive control (rifampin) are 1 μM and 10 μM, and the final concentration of the negative control (propranolol) is 10 μM. The final concentrations of the test compound are 10, 1, 0.1 μM or 30, 10, 3, 1, 0.3, 0.1 μM. The final concentration of DMSO is 0.1%.

[0362] 2) Remove the cell culture plate from the incubator and add 25 nL of positive and negative control drugs or test compound stock solutions directly using Echo, setting up three replicates for each concentration. Place the cell culture plate back into the incubator and continue incubation for 48 hours.

[0363] 3) Before starting experiments using the substrate, check cell morphology and monolayer integrity to ensure that the monolayer has acceptable research quality.

[0364] 3. Quantitative detection of PXR activation

[0365] 1) After 48 hours of drug treatment, the culture can be used for quantitative detection of PXR activation.

[0366] 2) CellTiter-Fluor TM Cell viability assay kit and One-Glo Luciferase reagent were equilibrated to room temperature. GF-AFC substrate (10 μL) was added to Assay Buffer (10 mL) to form a 2X reagent, which was then diluted with 10 mL PBS to form a 1X reagent. ONE-Glo Luciferase substrate was added to ONE-Glo Luciferase Assay Buffer.

[0367] 3) Remove the culture plate from the incubator, discard the culture medium, and add 1X CellTiter-Fluor TM Pour the reagent into the sample loading tank, add 25 μL of reagent to each well of the culture plate using a pipette, and then incubate in an incubator for 30 minutes.

[0368] 4) Remove the cell culture plate from the incubator, let it cool slightly to room temperature, and measure the fluorescence value using a fully automated quantitative microplate reader. The excitation light is 400nm and the emission light is 505nm.

[0369] 5) Pour ONE-Glo reagent into the sample loading tank, add 25 μL to each well, gently mix the plate, incubate at room temperature for 5 minutes, and measure the luminescence value.

[0370] 4. Data Analysis

[0371] All data was calculated using Microsoft Excel.

[0372] 1) The activity of luciferase is represented by RFU / RLU, where RLU is the average luminescence intensity value of three parallels for each concentration of each compound, and RFU is the average fluorescence intensity value of three parallels for each concentration of each compound.

[0373] The activation fold of mRNA is calculated using the following formula: Fold activation = (RLU test / RFU test) / (RLUvehicle / RFUvehicle)

[0374] 2) The cell viability percentage of the compound is calculated using the following formula: Cell Viability% = (RFUtest / RFUvehicle) × 100

[0375] 3) The percentage relative to the positive control is calculated using the following formula: Percent of positive control (%) = (Fold activation test / Fold activation Positive control) × 100

[0376] The experimental results are shown in Table 2:

[0377] Table 2

[0378] Conclusion: The compounds of the present invention, such as the diglucamine salt of compound 2, have no significant CYP3A4 induction effect at 0.1 μM, and have a lower risk of drug-drug interactions.

[0379] Biological Test Example 7: Solubility Test

[0380] Weigh approximately 1.0 mg of each compound and control compound for each test to determine solubility (aliquots into three separate 1.5 mL glass vials). One vial for each compound is used to prepare the standard, and the other two vials are used for parallel solubility determinations. Place each compound sequentially into its corresponding 96-well plate holder: one holder for standards and the other for the test sample. Based on the weighed mass, use a pipette to add an appropriate amount of matrix to each vial of the solubility plate. Add a stir bar to each vial and seal the vial with a molded PTFE / silicone stopper. Transfer the solubility plate to an Eppendorf Thermomixer Comfort plate shaker and shake at 25°C and 1100 RPM for 24 hours. After 24 hours, remove the stir bar with a large magnet, transfer the sample from the solubility plate to a filter plate, and filter all compounds using a vacuum manifold. Dilute the filtrate 1000-fold: Take 10 μL of the filtrate, add 10 μL of DMSO and 980 μL of a 1:1 water-acetonitrile mixture containing internal standards (IS, containing 100 nM tolbutamide, 200 nM labetalol, and 100 nM ketoprofen), vortex to mix, then take another 100 μL of the diluted solution and add 900 μL of a 1:1 water-acetonitrile mixture containing the same internal standards. Transfer 200 μL of the final diluted solution to a new 96-well plate for LC-MS / MS analysis. Adjust the dilution factor according to the solubility measurement and LC-MS signal response.

[0381] For the standards, calculate the required volume of DMSO based on the weighed mass to achieve a concentration of 1.0 mg / mL for each standard, and add the corresponding volume of DMSO to each standard vial using a pipette. Seal the standard vials with the 96-well plate cap and place the plate in an Eppendorf Thermomixer Comfort plate shaker, shaking at 25°C and 1100 rpm for 5 minutes. After 5 minutes, all standards will be completely dissolved. Transfer 10 μL from the 1.0 mg / mL DMSO standard plate to a new plate, add 10 μL of buffer and 980 μL of a 1:1 water-acetonitrile mixture containing internal standards (IS, containing 100 nM tolbutamide, 200 nM labetalol, and 100 nM ketoprofen) to obtain a 10 μg / mL standard solution. Transfer 100 μL from the 10 μg / mL standard plate to a new plate, add 900 μL of a 1:1 water-acetonitrile mixture containing the same internal standard, to obtain a standard solution with a final concentration of 1 μg / mL. Transfer 200 μL to a new 96-well plate for LC-MS / MS analysis. The standard concentration is adjusted based on the LC-MS signal response. Place the solubility sample plate and standard plate in an autosampler for analysis and evaluation by LC-MS / MS.

[0382] All calculations were performed using Microsoft Excel. Peak identification and quantification were performed using liquid chromatography-mass spectrometry (LC-MS), and the filtrate was compared with standards of known concentrations. The solubility calculation formula for the analyte and control compounds is as follows: Solubility = Standard concentration × Sample dilution factor × Sample peak area ratio / Standard peak area ratio

[0383] result:

[0384] Table 3. Solubility of Compounds (μg / mL)

[0385] Conclusion: The compounds of the present invention, such as the example compounds, have better solubility than control compound 1 and can be used for injection administration.

[0386] Biological Test Example 8: Effects of Compounds on TTX-R Sodium Channel Currents in Rat DRG Neurons

[0387] 1. Isolation of rat DRG neurons

[0388] 1.1 Laboratory Animals

[0389] In this project, we used acutely isolated rat DRG neurons. Male SD rats, 2-3 weeks old, SPF grade, were purchased from Spiefolk (Beijing) Biotechnology Co., Ltd., and all rats were fed for approximately 7 days before being used in the experiment.

[0390] 1.2 Solution Preparation

[0391] HBSS buffer: 145mM NaCl, 2.5mM KCl, 1mM MgCl2·6H2O, 2mM CaCl2·2H2O, 10mM D-Glucose, 10mM HEPES, pH adjusted to 7.4 with NaOH.

[0392] 1.3 DRG Separation Steps

[0393] (1) The experimental animals were weighed, and the SD rats were anesthetized with 0.1 mL / 10 g of 25% urethane and placed in a prone position. The skin was disinfected with 75% alcohol, and then the skin was cut open from the tail with surgical scissors to fully expose the spine and remove the muscle tissue on both sides of the spine.

[0394] (2) Next, the spine was removed from the coccyx to the cervical spine and placed in a 10cm culture dish containing HBSS buffer on an ice box. The upper 1 / 3 of the cross-section of the spine was cut off with scissors. After removing the spinal cord, the nerves connecting to the DRG were visible. Then, under a dissecting microscope, the DRG nodules of the L4-L6 segments of the lumbar spine were removed, and the nerves connecting to the DRG were cut off under a microscope.

[0395] (3) Cut the treated DRG nodules into small tissue pieces, aspirate them into 15 mL centrifuge tubes, centrifuge at 1000 rpm for 5 min, and discard the supernatant. Then add 5 mL of digestion solution (containing Trypsin + Collagenase) to the centrifuge tubes and place them in an incubator at 37℃ and 5% CO2 for about 20 min of digestion. Gently shake the centrifuge tubes every 5 min to resuspend the tissue. Use a pipette to blow and agitate the tissue pieces to promote digestion. During the digestion process, the tissue can be seen to gradually become filamentous. After two or three repeated operations, the filamentous material disappears and there are no obvious tissue pieces, indicating that digestion is complete.

[0396] (4) After complete tissue digestion, add 5 mL of DMEM medium containing 10% fetal bovine serum (FBS) and 1% penicillin / strep to terminate digestion, centrifuge again and discard the supernatant. Finally, add an appropriate amount of medium, gently resuspend the cells, and evenly add the cell suspension to 24-well plates with pre-coated poly-L-lysine cell spreaders. Let stand for about 2 hours until the cells adhere to the plate, then use them for electrophysiological experiments.

[0397] 2. Electrophysiological recording

[0398] 2.1 Record the liquids used

[0399] (1) Extracellular fluid: DRG-004-1

[0400] 22mM NaCl, 110mM Choline chloride, 0.8mM MgCl2·6H2O, 1.8mM CaCl2·2H2O, 5mM D-Glucose, 10mM HEPES, 0.1mM CdCl2, and NaOH were added to adjust the pH to 7.4; 300nM TTX was added to the external solution to block the TTX-S channel current.

[0401] (2) Intracellular fluid: DRG-004-2

[0402] 10mM CsCl, 5mM NaCl, 10mM HEPES, 2mM Mg-ATP, 135mM CsF, 5mM EGTA, CsOH adjust pH=7.2.

[0403] Extracellular fluid should be stored at 4°C and used within 2 weeks. Intracellular fluid, after preparation, should be aliquoted into 1 mL tubes and stored at -20°C. Freshly thawed intracellular fluid should be used daily for experiments. All intracellular fluid should be used within three months; otherwise, discard and prepare fresh.

[0404] 2.2 Patch clamp testing

[0405] The voltage stimulation protocol for whole-cell patch-clamp recording of Rat DRG TTX-R sodium currents is as follows: After whole-cell sealing, the cell voltage is clamped at -120 mV. The voltage is first stepped from -130 mV to -10 mV in 10 mV increments and held for 5 s, followed by a 0 mV depolarization pulse to obtain the half-inactivated voltage (Vhalf). The resting state and half-inactivated state of the sodium current are detected using a dual-pulse mode. First, a depolarization pulse (TP1) to 0 mV is applied for 50 ms to detect the resting sodium current. Then, the voltage is adjusted to Vhalf and held for 5 s, followed by restoring the voltage to -120 mV and holding for 20 ms to allow the unbound and inactivated channels to recover. A second depolarization pulse (TP2) to 0 mV is then applied for 50 ms to detect the half-inactivated sodium current. Finally, the voltage was restored to the clamping voltage of -120mV, and data was collected repeatedly at 20s intervals to observe the effect of the drug on the peak sodium current in the two different states.

[0406] Experimental data were acquired by an EPC 10 amplifier (HEKA) and stored in PatchMaster (HEKA) software.

[0407] The patch-clamp procedure begins by using a microelectrode puller to draw a glass capillary into a recording electrode. The electrode, filled with intracellular fluid, is then placed into a microelectrode holder. Next, a cell slide coated with cells is placed in a recording bath under an inverted microscope. Under the microscope, the microelectrode manipulator is manipulated to immerse the electrode in the extracellular fluid, and the electrode resistance (Rpip) is recorded. The electrode is then slowly brought into contact with the cell surface, and negative pressure is applied to create a GΩ high-resistance seal. Fast capacitance electrode compensation is then performed, and negative pressure is continued to rupture the cell membrane, establishing a whole-cell recording mode. Finally, slow capacitance cell compensation is performed, and experimental parameters such as series resistance (Rs) are recorded. No leakage compensation is applied.

[0408] Once the current amplitude in the cells stabilized in the solvent control, drug administration began, followed by sequentially increasing the concentration of the working solution of the test compound. Each concentration was applied until the current response reached equilibrium (within approximately 5 minutes), at which point the next concentration was introduced. All solutions were applied to the cell surface via a gravity perfusion system, while a peristaltic pump continuously replaced the fluid in the recording bath. Electrophysiological recording was performed at room temperature.

[0409] 2.3 Data Quality Control Standards

[0410] The following criteria are used to determine whether data is acceptable:

[0411] (1) Electrode resistance <5MΩ

[0412] (2) Sealing resistance > 1 GΩ

[0413] (3) Initial connection resistance <15MΩ

[0414] (4) Connection resistor ends <15MΩ

[0415] (5) The absolute value of the peak value of the starting current is >300pA

[0416] (6) The current does not show obvious spontaneous decay.

[0417] (7) At the same concentration, the difference in inhibition rate between repeated data is ≤15%.

[0418] 3. Data Analysis

[0419] Peak sodium current after each drug concentration was applied. compound ) and peak current (contrast current) control Normalize, then calculate the inhibition rate corresponding to each drug concentration, i.e., Inhibition% = (1 - (Peak current)) / (Peak current) compound ) / (Peak current control ))*100%, and calculate the mean (Mean), standard deviation (SD), and standard error (SE) for each drug concentration inhibition rate. The data are expressed as Mean±SE.

[0420] A four-parameter nonlinear fitting was performed on the concentration-inhibition rate curve. Absolute IC50 was then calculated. 50 and IC 90 The X values ​​can be obtained by interpolation, specifically when Y = 50 and Y = 90. Wherein, IC... 50 The half-inhibitory concentration, IC50 90 The concentration at which the inhibition rate is 90% is given, where X is the logarithm of the concentration and Y is the inhibition rate. IC50 50 and IC 90 The calculations and curve fitting were performed using GraphPad Prism software.

[0421] Conclusion: The compounds of this invention have a significant inhibitory effect on the TTX-R sodium channel current in rat DRG neurons, for example, the IC50 of compound 2 diglucamine salt. 50 <10nM,IC 90 <30 nM, IC50 of control compound VX-548 50 >60nM, IC 90 >600nM.

[0422] Biological Test Example 9: Solid Stability Evaluation

[0423] Samples were packaged in double-layered pharmaceutical-grade low-density polyethylene bags and placed under conditions of 60% ± 5% RH, 25℃ ± 2℃ or 75% ± 5% RH, 40℃ ± 2℃. Samples were taken on day 0 and day 10, and their HPLC purity was tested using high-performance liquid chromatography (HPLC).

[0424] HPLC conditions: Sample solution: Accurately weigh approximately 25 mg of sample, place in a 25 ml volumetric flask, dissolve and dilute to the mark with solvent, and mix well; Column: Octadecylsilane-bonded silica gel (Agilent Eclipse Plus C18, 4.6 mm × 150 mm, 3.5 μm); Mobile phase A: 0.05 mol / L ammonium acetate-acetonitrile (85:15); Mobile phase B: acetonitrile; Diluent: acetonitrile-water = 8:2; Detection wavelength: 248 nm; Flow rate: 1.0 ml / min; Column temperature: 35 ℃; Injection volume: 10 μL;

[0425] Washing procedure:

[0426] Calculation method: The result is obtained by area normalization.

[0427] The results are shown in Table 4:

[0428] Table 4

[0429] Conclusion: Compound 2 exhibits good solid stability and better drug-like properties.

[0430] Biological Test Example 10: Safety Evaluation

[0431] We evaluated the safety of compounds by screening for the functional activity of 90 targets to assess their agonistic or inhibitory effects on those targets.

[0432] 1. Experimental Methods

[0433] 1.1 Detection method for GPCR targets

[0434] 1.1.1. Gq-coupled receptor activity assay (FLIPR method)

[0435] 1) Cell plating: Digest and collect cells, resuspend and count them, and then seed them into 384-well cell plates. Then incubate the cell plates at 37°C in a 5% CO2 incubator for about 16-20 hours.

[0436] 2) Prepare the Assay Buffer according to the FLIPR Calcium 6 Assay Kit instructions. Melt 20× Component A to room temperature, dilute it with Assay Buffer to 1× loading buffer, and store at room temperature.

[0437] 3) Remove the culture medium from the cell plate, quickly add 20 μL of 1× loading buffer to each well, centrifuge at 1000 rpm for 1 minute at room temperature, and then incubate the cell plate at 37°C in the dark for 120 minutes.

[0438] 4) Prepare 5× positive control and test compound working solutions, and transfer 20 μL / well to a 384-well compound source plate.

[0439] 5) Simultaneously prepare the 2.5× agonist working solution required for the antagonistic mode and transfer 40 μL / well to the 384-well compound source plate.

[0440] 6) Place the cell plate, compound source plate, and pipette tip into the corresponding positions of the high-throughput real-time fluorescence detection system. Use FLIPR Tetra to add 5 μL of the diluted compound from step 4 into each experimental well, and collect data at a wavelength of 515 nm-575 nm, which is recorded as the activation mode reading.

[0441] 7) After data acquisition, use FLIPR to add 5 μL of the corresponding compound to each well and incubate at 37°C in the dark for 15 minutes.

[0442] 8) Place the cell plate, compound source plate, and pipette tip into the corresponding positions of the high-throughput real-time fluorescence detection system. Use FLIPR Tetra to add 20 μL of the agonist diluted in step 5 into each experimental well, and collect data at a wavelength of 515 nm-575 nm, which is recorded as the antagonistic mode reading.

[0443] The targets detected using this experiment are: M1, M2, M3, 5-HT2A, 5-HT2B, H1, α1A, α2A, α1B, α2B, 5-HT4A, M4, NK1, NK2, M5 and 5-HT2C.

[0444] 1.1.2. Detection of Gq-coupled receptor activity (IP-One method)

[0445] 1) Prepare 1×Stimulation Buffer according to the IP-One-Gq kit instructions.

[0446] 2) Dilute with DMSO or the corresponding solvent. Prepare positive controls at a concentration of 100× for activation modes V1a, CCK2, and CCK1. Prepare positive controls at a concentration of 1000× for ETB and ETA (prepare positive controls at a concentration of 1000× for antagonistic modes). Prepare working solution of test substance at a concentration of 1000×. Transfer the compound to a 384 reaction plate using ECHO665. Seal the plate with sealing film and centrifuge at 1000 rpm for 1 minute. Set aside for use.

[0447] 3) Cells were digested with trypsin, centrifuged at 1000 rpm for 5 minutes at room temperature, resuspended in 1×Stimulation Buffer, counted, and seeded into 384 reaction plates. After centrifugation at 1000 rpm for 1 minute at room temperature, the plates were incubated at 37°C for 70 minutes. (Antagonist mode: After incubating cells for 10 minutes, agonists were added using ECHO655, and the plates were then incubated at 37°C for 1 hour to induce IP1 production).

[0448] 4) Dilute IP1-d2 to 7× working concentration with Lysis & Detection Buffer, and then add it to the corresponding experimental well.

[0449] 5) Dilute Anti-IP1-Cryptate to 7× working concentration with Lysis & Detection Buffer, then add it to the corresponding experimental wells, centrifuge at 1000 rpm for 1 minute at room temperature, and incubate at room temperature for 1 hour.

[0450] 6) After incubation, use an ELISA reader to detect the readings at 665nm and 620nm under excitation at a wavelength of 330nm.

[0451] The targets detected using this experiment are: V1a, CCK1, CCK2, ETB, and ETA.

[0452] 1.1.3. Detection of Gs-coupled receptor activity (HTRF method)

[0453] 1) Prepare 1×Stimulation Buffer according to the LANCE Ultra cAMP kit instructions.

[0454] 2) Dilute with DMSO or the corresponding solvent to prepare a working solution of positive control and test substance at a concentration of 1000×. Transfer the compound to a 384 reaction plate using ECHO665, seal with sealing film, and centrifuge at 1000 rpm for 1 minute.

[0455] 3) Cells were trypsinized, centrifuged at 1000 rpm for 1 minute at room temperature, resuspended in 1×Stimulation Buffer, counted, and seeded into 384-well plates. After centrifugation at 1000 rpm for 1 minute, the cells were incubated at 37°C for 40 minutes. (Antagonist mode: After incubation for 10 minutes with cells, ECHO was added as an agonist, followed by incubation at 37°C for 30 minutes to induce cAMP production.)

[0456] 4) Dilute Eu-cAMP to 4× working concentration with detection buffer and then add it to the corresponding wells of the 384 reaction plate.

[0457] 5) Dilute ULight-anti-cAMP to 4× working concentration with detection buffer, then add it to the corresponding 384 reaction plate wells, centrifuge at 1000 rpm for 1 minute at room temperature, and incubate at room temperature for 1 hour.

[0458] 6) After incubation, use an ELISA reader to detect the readings at 665nm and 620nm under excitation at a wavelength of 330nm.

[0459] The targets detected using this experiment are: β1, β2, D1, H2, A2A, EP2, MC4 and 5-HT7A.

[0460] 1.1.4. Gi-coupled receptor activity assay (HTRF method)

[0461] 1) Prepare 1×Stimulation Buffer according to the LANCE Ultra cAMP kit instructions.

[0462] 2) Dilute with DMSO or the corresponding solvent to prepare a positive control and test solution of 1000× test concentration, and prepare 1000× Forskolin (2000× Forskolin and agonist for antagonist mode). Transfer the compound to a 384 reaction plate using ECHO665, seal with sealing film, and centrifuge at 1000 rpm for 1 minute for later use.

[0463] 3) Cells were digested with trypsin, centrifuged at 1000 rpm for 1 minute at room temperature, resuspended in 1×Stimulation Buffer, counted, and seeded into 384-well plates. After centrifugation at 1000 rpm for 1 minute, the cells were incubated at 37°C for 30 minutes. After incubation for 10 minutes, Forskolin was added using ECHO (antagonist mode: 5 nL of a mixture of Forskolin and agonist was added using ECHO, followed by incubation at 37°C for 30 minutes to induce cAMP production).

[0464] 4) Dilute Eu-cAMP to 4× working concentration with detection buffer and then add it to the corresponding wells of the 384 reaction plate.

[0465] 5) Dilute ULight-anti-cAMP to 4× working concentration with detection buffer, then add it to the corresponding 384 reaction plate wells, centrifuge at 1000 rpm for 1 minute at room temperature, and incubate at room temperature for 1 hour.

[0466] reagent concentration

[0467] 6) After incubation, use an ELISA reader to detect the readings at 665nm and 620nm under excitation at a wavelength of 330nm.

[0468] The targets detected using this experiment are: CB1, CB2, 5-HT1A, 5-HT1B, D2S, DOR, KOR, and MOR.

[0469] 1.2. Detection methods for ion channel targets

[0470] 1.2.1. Manual Patch Clamp Testing Method

[0471] Manual Patch Clamp Testing Method

[0472] I Ks Electrophysiological assays were performed using manual patch-clamp techniques. Experimental data were acquired using an EPC-10 amplifier (HEKA) and stored in PatchMaster (HEKA) software. Under an inverted microscope, the microelectrode manipulator was used to bring the recording electrodes into contact with the cells, and negative pressure was applied to aspirate and form a GΩ seal. After GΩ sealing, rapid capacitance compensation was performed, followed by continued negative pressure to rupture the cell membrane, establishing a whole-cell recording mode. Slow capacitance compensation was then performed, and membrane capacitance and series resistance were recorded. No leakage compensation was applied. A coverslip containing cells was placed in the recording bath within the inverted microscope. Test compounds and external solutions without the compounds were sequentially perfused through the recording bath using gravity, acting on the cells. Fluid exchange was performed using a vacuum pump during recording. At least two cells were independently and repeatedly analyzed. All manual electrophysiological experiments were performed at room temperature.

[0473] 1.2.2. FLIPR Detection Method

[0474] 1) Cell plating: Digest and collect cells, resuspend and count them, and then seed them into 384-well cell plates. Then incubate the cell plates at 37°C in a 5% CO2 incubator for about 16-20 hours.

[0475] 2) Prepare the Assay Buffer according to the FLIPR Calcium 6 Assay Kit instructions. Melt 20× Component A to room temperature, dilute it with Assay Buffer to 1× loading buffer, and store at room temperature.

[0476] 3) Remove the culture medium from the cell plate, quickly add 1× loading buffer to each well, centrifuge at 1000 rpm for 1 minute at room temperature, and then incubate the cell plate at 37°C in the dark.

[0477] (GABA A (α1β2γ2) and GlyRα1β were incubated for 30 minutes, SK1 and KCNQ2 / 3 were incubated for 1 hour, and TRPV1, 5-HT3A, P2X3, NR1 / NR2B, nAChRα1β1δε, nAChRα4β2, Cav1.2, and Cav2.2 were incubated for 2 hours.

[0478] 4) Prepare 5× positive control and test compound working solutions, and transfer 20 μL / well to a 384-well compound source plate.

[0479] 5) Simultaneously prepare the 2.5× agonist working solution required for the antagonistic mode and transfer 40 μL / well to the 384-well compound source plate.

[0480] 6) Place the cell plate, compound source plate, and pipette tip into the corresponding positions of the high-throughput real-time fluorescence detection system. Use FLIPR Tetra to add the diluted compound from step 4 into each experimental well and collect data at the corresponding wavelength, which is recorded as the activation mode reading.

[0481] 7) After data acquisition, use FLIPR to add the corresponding compound to each well and incubate at 37°C in the dark for 15 minutes.

[0482] 8) Place the cell plate, compound source plate, and pipette tip into the corresponding positions of the high-throughput real-time fluorescence detection system. Use FLIPR Tetra to add the diluted agonist from step 5 into each experimental well and collect data at the corresponding wavelength, which is recorded as the antagonistic mode reading.

[0483] The targets detected using this experiment include: GABA. A (α1β2γ2), TRPV1, 5-HT3A, P2X3, nAChRα4β2, nAChRα1β1δε, KCNQ2 / 3, SK1, GlyRα1β, NR1 / NR2B, Cav1.2, Cav2.2. Among them GABA A(α1β2γ2) and GlyRα1β were detected using a membrane potential assay kit; TRPV1, 5-HT3A, P2X3, NR1 / NR2B, nAChRα1β1δε, nAChRα4β2, Cav1.2, and Cav2.2 were detected using a calcium flux assay kit; SK1 and KCNQ2 / 3 were detected using a potassium flux assay kit. The excitation light for membrane potential detection was 510-545 nm, and the emission light was 565-625 nm; the excitation light for calcium flux detection was 470-515 nm, and the emission light was 515-575 nm; the excitation light for potassium flux detection was 470-495 nm, and the emission light was 515-575 nm.

[0484] 1.2.3. hERG target detection method (FLIPR method)

[0485] 1) Resuscitate or digest cells, resuspend and count them, and then seed them into 384-well cell culture plates. Then incubate the 384-well cell culture plates at 37°C and 5% CO2 for about 16-20 hours.

[0486] 2) Prepare 1×buffer according to the kit instructions, and then use 1×buffer to prepare 2×dye for later use.

[0487] 3) Centrifuge at 300 rpm for 5 seconds at room temperature to remove the culture medium from the 384-well cell plate, and immediately add 20 μL of 1× buffer.

[0488] 4) Take 20 μL of the dye prepared in step 2 and add it to the corresponding 384-well cell plate experimental wells, and incubate at 37°C in the dark for 1 hour.

[0489] 5) Prepare 5× positive control and test intermediate solutions and transfer them to the corresponding 384-well compound intermediate source plates.

[0490] 6) Prepare 6× agonist intermediate solution and transfer it to the corresponding 384-well positive control intermediate solution source plate.

[0491] 7) Using a FLIPR instrument, add 10 μL of the compound prepared in step 5 to each well of a 384-well cell plate and record data for 5 minutes; then incubate for 15 minutes. Next, add 10 μL of the agonist prepared in step 6 to each well of the 384-well cell plate and record data for 5 minutes. Excitation light is 470-495 nm, and emission light is 515-575 nm.

[0492] 1.3. Detection methods for enzyme-related targets

[0493] 1.3.1. Determination of the inhibitory effect of the compound on the activities of PDE3A and PDE4D2 enzymes

[0494] 1) Dilute the test sample stock solution with DMSO, transfer 25 nL of the 200× test concentration compound solution to a 384 reaction plate, and centrifuge at 1000 rpm for 1 minute at room temperature.

[0495] 2) Add 2.5 μL of 2×PDE to each detection well and centrifuge at 1000 rpm for 1 minute.

[0496] 3) Add 2.5 μL of 2×FAM-cAMP to each detection well, centrifuge at 1000 rpm for 1 minute, and incubate at room temperature for 60 minutes.

[0497] 4) Add 15 μL of binding agent mixture to each detection well, centrifuge at 1000 rpm for 1 minute, and incubate at room temperature for 60 minutes.

[0498] 5) Use an ELISA reader to read the fluorescence polarization signals at 485nm and 520nm.

[0499] 1.3.2. Determination of the inhibitory effects of the compound on the activities of PDE3B, PDE5A1, and PDE6C enzymes

[0500] 1) Dilute the test compound stock solution with DMSO, transfer 25 nL of the 200× test concentration compound solution to a 384-well container, and centrifuge at 1000 rpm for 1 minute at room temperature.

[0501] 2) Transfer 2.5 μL of 2×PDE enzyme to a 384 reaction plate and centrifuge at 1000 rpm for 1 minute at room temperature.

[0502] 3) Transfer 2.5 μL of 2×FAM-Cyclic-3,5-AMP / GMP to a 384 reaction plate, centrifuge at 1000 rpm for 1 minute at room temperature, and incubate at 25°C for 1 hour.

[0503] 4) Transfer 15 μL of binding agent mixture to a 384 reaction plate, centrifuge at 1000 rpm for 1 minute at room temperature, and incubate at 25°C for 1 hour.

[0504] 5) Use an ELISA reader to read the fluorescence polarization signals at 485nm and 520nm.

[0505] 1.3.3. Determination of the inhibitory effects of the compound on the activities of ACE, ACE2, BACE-1, MMP1 and MMP2 enzymes

[0506] 1) Dilute the test compound stock solution with DMSO, transfer 50 nL of the 200× test concentration compound solution to a 384-well container, and centrifuge at 1000 rpm for 1 minute at room temperature.

[0507] 2) Transfer 5 μL of 2× enzyme to a 384 reaction plate, centrifuge at 1000 rpm for 1 minute at room temperature, and incubate at 25°C for 15 minutes.

[0508] 3) Transfer 5 μL of 2× substrate to a 384 reaction plate, centrifuge at 1000 rpm for 1 minute at room temperature, and incubate at 25°C for 90 minutes.

[0509] 4) Read the fluorescence value FI (ex 320±15 / em 405±20) using an ELISA reader.

[0510] 1.3.4. Determination of the inhibitory effects of the compound on the activities of Caspase 3 and Cathepsin G enzymes

[0511] 1) Dilute the test compound stock solution with DMSO, transfer 50 nL of the 200× test concentration compound solution to a 384-well container, and centrifuge at 1000 rpm for 1 minute at room temperature.

[0512] 2) Transfer 5 μL of 2× enzyme to a 384 reaction plate, centrifuge at 1000 rpm for 1 minute at room temperature, and incubate at 25°C for 15 minutes.

[0513] 3) Transfer 5 μL of 2× substrate to a 384 reaction plate, centrifuge at 1000 rpm for 1 minute at room temperature, and incubate at 25°C for 180 minutes.

[0514] 4) Read the fluorescence value FI (ex 360±15 / em 450±20) using an ELISA reader.

[0515] 1.3.5. Determination of the inhibitory effect of compounds on COX1 and COX2 enzyme activities (FI method)

[0516] 1) Dilute the test sample stock solution with DMSO.

[0517] 2) Use Echo to transfer 50 nL of the 200× test concentration compound solution to the reaction plate and centrifuge at 1000 rpm for 1 minute at room temperature.

[0518] 3) Transfer 2.5 μL of 4× enzyme to the reaction plate and centrifuge at 1000 rpm for 1 minute at room temperature.

[0519] 4) Add 2.5 μL of 4×Hemin mix to the reaction plate, centrifuge at 1000 rpm for 1 minute at room temperature, and incubate at 25°C for 10 minutes.

[0520] 5) Transfer 2.5 μL of 4×ADHP to a 384 reaction plate and centrifuge at 1000 rpm for 1 minute at room temperature.

[0521] 6) Transfer 2.5 μL of the 4×AA (arachidonic acid) and KOH mixture to a 384 reaction plate and centrifuge at 1000 rpm for 1 minute at room temperature.

[0522] 7) Read the fluorescence value on the microplate reader (excitation wavelength 528±15nm, emission wavelength 587±20nm).

[0523] 1.3.6. Determination of the inhibitory effect of compounds on kinase target activity (HTRF method)

[0524] 1) Dilute the test sample stock solution with DMSO.

[0525] 2) Use Echo to transfer 25 nL of the compound solution at 200× test concentration to the target plate and centrifuge at 1000 rpm for 1 minute at room temperature.

[0526] 3) Add 2.5 μL of 2×kinase / metal ion solution to the plate and incubate at 25°C for 10 minutes.

[0527] 4) Add 2.5 μL of 2× substrate / ATP solution to the plate and incubate at 25°C for 50 minutes.

[0528] 5) Prepare XL665 and antibody detection reagents using detection buffer.

[0529] 6) Add 5 μL of kinase detection reagent to the plate and incubate at 25°C for 60 minutes.

[0530] 7) Read the fluorescence signals at 620nm (Cryptate) and 665nm (XL665) using an ELISA reader.

[0531] The target sites suitable for this method are CaMK2α and LCK.

[0532] 1.3.7. Determination of the inhibitory effect of compounds on kinase target activity (ADP-Glo ​​method)

[0533] 1) Dilute the test sample stock solution with DMSO.

[0534] 2) Use Echo to transfer 20 nL of the 200× test concentration compound solution to the target plate and centrifuge at 1000 rpm for 1 minute at room temperature.

[0535] 3) Add 2 μL of 2×kinase / metal ion solution to the plate and incubate at 25°C for 10 minutes.

[0536] 4) Add 2 μL of 2× substrate / ATP solution to the plate and incubate at 25°C for 60 minutes.

[0537] 5) Add 4 μL of ADP-Glo ​​detection reagent to the plate and incubate at 25°C for 40 minutes.

[0538] 6) Add 8 μL of Detection reagent to the plate and incubate at 25°C for 40 minutes.

[0539] 7) Read the chemiluminescence signal using an enzyme-linked immunosorbent assay (ELISA) reader.

[0540] The targets suitable for this method include ROCK1, INSR, VEGFR2, ABL1, VEGFR1, Fyn, ZAP70, CDK2, ERK2, GSK3α, GSK3β, p38α, and PKCα.

[0541] 1.3.8. Determination of the inhibitory effect of the compound on AChE enzyme activity

[0542] 1) Dilute the test sample stock solution with DMSO.

[0543] 2) Use an Echo to transfer 100 nL of the 200× test concentration compound solution to the reaction plate and centrifuge at 1000 rpm for 1 minute at room temperature.

[0544] 3) Add 10 μL of 2×AchE solution to the reaction plate, centrifuge at 1000 rpm for 1 minute at room temperature, and incubate at 25°C for 10 minutes.

[0545] 4) Transfer 10 μL of the 2×(ATC, DTNB) mixture to a reaction plate, centrifuge at 1000 rpm for 1 minute at room temperature, and incubate at 25°C for 90 minutes.

[0546] 5) Read the value using an ELISA reader (OD: 405nm).

[0547] 1.3.9. Determination of the inhibitory effect of the compound on MAOA / B enzyme activity

[0548] 1) Dilute the test sample stock solution with DMSO.

[0549] 2) Use an Echo to transfer 25 nL of the 200× test concentration compound solution to the reaction plate and centrifuge at 1000 rpm for 1 minute at room temperature.

[0550] 3) Add 2.5 μL of 2×MAO enzyme solution to the reaction plate, centrifuge at 1000 rpm for 1 minute at room temperature, and incubate at 25°C for 10 minutes.

[0551] 4) Transfer 2.5 μL of 2×MAO-Substrate solution to the reaction plate, centrifuge at 1000 rpm for 1 minute at room temperature, and incubate at 25°C for 60 minutes.

[0552] 5) Transfer 5 μL of MAO-Glo, centrifuge at 1000 rpm for 1 minute at room temperature, and incubate at 25°C for 30 minutes.

[0553] 6) Read the values ​​on the ELISA reader (Luminescence).

[0554] 1.4. Detection methods for nuclear receptor targets

[0555] 1) Day 1: Digest cells with 1×0.25% Trypsian-EDTA, then resuspend and count cells in DMEM medium without phenol red containing 5% carbon-adsorbed serum, and seed them into 384-well white blood cell culture plates. Add the compound to each well and incubate overnight, then add DMEM without phenol red to each well (antagonistic mode: add the compound to each well and incubate for half an hour, then add the agonist solution and continue incubation overnight).

[0556] 2) Day 2: 22 hours after drug administration, luciferase activity was detected using the Bright-Lite Luciferase Assay System. The 384-well plate containing the test cells was removed from the incubator, Bright-Lite Reagent was added, and the cells were shaken to allow for complete lysis. The luminescence signal value was then read using a multi-mode microplate reader.

[0557] The targets detected using this experiment are: AR, GR, and ERα.

[0558] 1.5. Detection method for transporter target points

[0559] 1) Digest and collect cells, count them, and then inoculate them in 384-well plates and culture them overnight.

[0560] 2) Prepare 1× buffer solution according to the kit instructions.

[0561] 3) Prepare positive compound and test substance 2× working solution.

[0562] 4) Centrifuge to remove the culture medium from the 384-well plate. Add the diluted compound from step 3 to the corresponding experimental wells, centrifuge, and incubate at 37°C for 30 minutes.

[0563] 5) Prepare the test reagent with HBSS solution, add an equal volume of test reagent to each well, centrifuge, and incubate at 37°C for 1 hour.

[0564] 6) After incubation, use a multi-functional microplate reader to detect the reading at 520±20nm, with an excitation light of 440±20nm.

[0565] The targets detected using this experiment are: DAT, NET, and 5-HTT.

[0566] 2. Calculation method:

[0567] V1a, CCK1, CCK2, H1, ETA, ETB, α1A, α1B, α2A, α2B, 5-HT4A, NK1, NK2, M1, M2, M3, M4, M5, 5-HT2A, 5-HT2B, 5-HT2C, β1 , β2, D1, H2, A2A, EP2, MC4, 5-HT7A, CB1, CB2, 5-HT1A, 5-HT1B, D2S, DOR, KOR, MOR, AR, GR, Erα, 5-HT3A, TRPV1, GABA A (α1β2γ2), KCNQ2 / 3, nAChRα1β1δε, SK1, P2X3, GlyRα1β, NR1 / NR2B, nAChRα4β2, Cav2.2, Nav1.5, hERG, Cav1.2 target: Compound activation rate (%Act) = (signal 化合物 -Signal 阴性对照 ) / (Signal 阳性对照 -Signal 阴性对照 )*100; Compound inhibition rate (%Inh) = (signal) 化合物 -Signal 阴性对照 ) / (Signal 阳性对照 -Signal 阴性对照 )*100;

[0568] DAT, NET, 5-HTT, COX1, COX2, MAOA, PDE3A, PDE3B, PDE4D2, AchE, INSR, PDE5A1, PDE6C, Caspase 3, Cathepsin G, ACE, ACE2, BACE-1, MMP1, MMP2, PKCα, MAOB, ROCK1, VEGFR2, Fyn, ERK2, p38α (SAPK2A), Flt-1 (VEGFR1), CaMK2α, ABL1, CDK2, GSK3α, GSK3β, ZAP70, and LCK targets: Compound inhibition rate (%Inh) = (signal) 化合物 -Signal 阴性对照 ) / (Signal 阳性对照 -Signal 阴性对照 )*100;

[0569] The IC50 of the positive control drug was calculated using the nonlinear fitting formula of XLfit 5.5.0.5. 50 or EC 50 : Y=Bottom+(Top-Bottom) / (1+10^((LogIC 50 -X)×HillSlope)) Y=Bottom+(Top-Bottom) / (1+10^((LogEC 50-X)×HillSlope))

[0570] I Ks Data Analysis:

[0571] First, the current after each drug concentration was applied and the current in the blank solvent treatment group were normalized. Then calculate the inhibition rate corresponding to each drug concentration. Calculate the mean and standard error of the inhibition rate for each concentration.

[0572] Conclusion: The compounds of the present invention have good safety. For example, the diglucamine salt of compound 2 has no obvious activating or inhibiting effect on 90 target sites at 10 μM, and the activating or inhibiting rate is less than 50%.

Claims

A compound or its stereoisomers, tautomers, racemates, or pharmaceutically acceptable salts, wherein the compound is selected from compounds of general formula (I) or general formula (I-1). X is selected from O, S, -S(O)- or -S(O)2-; Q is selected from Q1 is selected from C 6-10 Aryl, 5- to 10-membered heteroaryl, C 5-10 Carbon rings, 5- to 10-membered heterocycles or The aryl, heteroaryl, carbocyclic, or heterocyclic group is optionally surrounded by 1 to 5 R groups. q replace; R is selected from C 1-6 alkyl, -C(=O)R q1 -P(=O)R q1 R q2 -C 1-4 Alkylene-OP(=O)R q1 R q2 -C(O)OC 1-4 Alkylene-OP(=O)R q1 R q2 -S(=O)2R q1 C 3-6 Carbocyclic, 4- to 7-membered heterocyclic, wherein the alkyl, alkylene, carbocyclic or heterocyclic is optionally surrounded by 1 to 4 R... k replace; R q1 R q2 Each is independently selected from H, OH, and C. 1-6 Alkyl, C 1-6 Alkoxy, C 3-6 Carbon ring, -OC 3-6 Carbocyclic, 4- to 7-membered heterocyclic, wherein the alkyl, alkoxy, carbocyclic or heterocyclic is optionally surrounded by 1 to 4 R... k replace; As an option, R q1 R q2 Direct connection forms a 4- to 7-membered heterocycle, wherein the heterocycle is optionally bounded by 1 to 4 R... k replace; R q3 R q4 Each is independently selected from H or arbitrarily selected by 1 to 4 Rs. k One of the following groups is substituted: C 1-6 Alkyl, -C 1-6 Alkylene -NH2, -C 1-6 Alkylene -C(=O)NH2, -C 1-6 Alkylene -NHC(=NH)NH2, -C 1-6 Alkylene -NHC(=O)NH2、-C 1-6 alkylene -OH, -C 1-6 Alkylene -SH, -C 1-6 Alkylene -CO2H, -C 1-6 Alkylene-C 3-10 Carbon ring, C 3-10 Carbon ring, -C 1-6 Alkylene-3 to 10-membered heterocycles, 3 to 10-membered heterocycles; As an option, R q3 R q4 Direct connection forms C 3-6 Carbon rings or 4- to 7-membered heterocycles, wherein the carbon rings or heterocycles are optionally surrounded by 1 to 4 R... k replace; B is selected from C. 6-10 Aryl, 5- to 10-membered heteroaryl, C 5-10 Carbocyclic rings, 5- to 10-membered heterocyclic rings, wherein the aryl, heteroaryl, carbocyclic, or heterocyclic rings are optionally surrounded by 1 to 5 R groups. B replace; R 1 R 2 R 3 R 4 R 5 Each is independently selected from H, halogens, CN, OH, and C. 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, -OC 1- 6-alkyl, -SC 1-6 Alkyl, C 3-6 Carbocyclic, 3- to 7-membered heterocyclic, wherein the alkyl, alkenyl, alkynyl, carbocyclic or heterocyclic is optionally surrounded by 1 to 4 R... k replace; As an option, R 1 R 4 Direct connection forms C 3-6 Carbon rings or 4- to 7-membered heterocycles, wherein the carbon rings or heterocycles are optionally surrounded by 1 to 6 R... k replace; R q R B Each is independently selected from H, deuterium, =O, halogen, CN, OH, NH2, NHC 1-6 Alkyl, N(C) 1-6 Alkyl)2, C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, -OC 1-6 Alkyl, -SC 1-6 Alkyl, C 3-7 Carbon ring, -OC 3-7 Carbon rings, 3- to 7-membered heterocycles, or -P(=O)R q1 R q2 The alkyl, alkenyl, alkynyl, carbocyclic, or heterocyclic group is optionally surrounded by 1 to 4 R groups. k replace; R k Each is independently selected from deuterium, =O, halogen, CN, OH, NH2, -C(=O)NH2, NHC 1-6 Alkyl, N(C) 1-6 Alkyl)2, C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, -OC 1-6 Alkyl, -SC 1-6 Alkyl, -OC 3-6 Carbocyclic rings, -O-3 to 7-membered heterocycles, -NH-C 3- 6-membered carbon rings, -NH-3 to 7-membered heterocycles, -C 1-4 Alkylene-C 3-6 Carbon ring, -C 1-4 Alkylene-3 to 7-membered heterocycles, C 3-6 The carbon ring, 3 to 7-membered heterocycle, wherein the alkyl, alkylene, alkenyl, alkynyl, carbon ring, or heterocycle is optionally selected from deuterium, halogen, =O, CN, OH, NH2, C 1-6 Alkyl, C 1-6 The alkoxy group is replaced by a substituent. According to claim 1, the compound or its stereoisomers, tautomers, racemates, or pharmaceutically acceptable salts thereof, R is selected from C 1-4 alkyl, -C(=O)R q1 -P(=O)R q1 R q2 -C 1-2 Alkylene-OP(=O)R q1 R q2 -C(O)OC 1-2 Alkylene-OP(=O)R q1 R q2 -S(=O)2R q1 C 3-6 Carbocyclic, 4- to 7-membered heterocyclic, wherein the alkyl, alkylene, carbocyclic or heterocyclic is optionally surrounded by 1 to 4 R... k replace; R q1 R q2 Each is independently selected from H, OH, and C. 1-4 Alkyl, C 1-4 Alkoxy, C 3-6 Carbon ring, -OC 3-6 Carbocyclic, 4- to 7-membered heterocyclic, wherein the alkyl, alkoxy, carbocyclic or heterocyclic is optionally surrounded by 1 to 4 R... k replace; As an option, R q1 R q2 Direct connection forms a 4- to 7-membered heterocycle, wherein the heterocycle is optionally bounded by 1 to 4 R... k replace; R q3 R q4 Each is independently selected from H or arbitrarily selected by 1 to 4 Rs. k One of the following groups is substituted: C 1-4 Alkyl, -C 1-4 Alkylene -NH2, -C 1-4 Alkylene -C(=O)NH2, -C 1-4 Alkylene -NHC(=NH)NH2, -C 1-4 Alkylene -NHC(=O)NH2、-C 1-4 alkylene -OH, -C 1-4 Alkylene -SH, -C 1-4 Alkylene -CO2H, -C 1-4 Alkylene-C 6-10 Aryl, C 6-10 Aryl, -C 1-4 alkylene-5 to 10-membered heteroaryl, 5 to 10-membered heteroaryl, C 3-6 cycloalkyl, 4- to 7-membered heterocyclic groups, -CH2-C 3-6 Cycloalkyl groups, -CH2-4 to 7-membered heterocyclic groups; As an option, R q3 R q4 Direct connection forms C 3-6 Cycloalkyl or 4- to 7-membered heterocycloalkyl, wherein the cycloalkyl or heterocycloalkyl is optionally surrounded by 1 to 4 R... k replace; Q1 is selected from phenyl, benzo[C] 4-6 Carbocyclic, benzo[4-6] heterocyclic, 5-6 fused heteroaryl, 5-6 fused heterocyclic, 8-10 fused heteroaryl or Q1 is arbitrarily selected by 1 to 4 Rs q replace; B is selected from phenyl, benzo[C] 4-6 Carbocyclic, benzo[4- to 6-membered heterocyclic, 5- to 6-membered heteroaryl, 8- to 10-membered fused-ring heteroaryl, wherein B is optionally surrounded by 1 to 4 R[]. B replace; R 1 R 2 R 3 R 4 R 5 Each is independently selected from H, halogens, CN, OH, and C. 1-4 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl group, -OC 1-4 Alkyl, -SC 1-4 Alkyl, C 3-6 Carbocyclic, 3- to 7-membered heterocyclic, wherein the alkyl, alkenyl, alkynyl, carbocyclic or heterocyclic is optionally surrounded by 1 to 4 R... k replace; As an option, R 1 R 4 Direct connection forms C 3-6 Carbon rings or 4- to 7-membered heterocycles, wherein the carbon rings or heterocycles are optionally surrounded by 1 to 4 R... k replace; R q R B Each is independently selected from H, deuterium, =O, halogen, CN, OH, NH2, NHC 1-4 Alkyl, N(C) 1-4 Alkyl)2, C 1-4 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl group, -OC 1-4 Alkyl, -SC 1-4 Alkyl, C 3-7 Carbon ring, -OC 3-7 Carbon rings, 3- to 7-membered heterocycles, or -P(=O)R q1 R q2 The alkyl, alkenyl, alkynyl, carbocyclic, or heterocyclic group is optionally surrounded by 1 to 4 R groups. k replace; R k Each is independently selected from deuterium, =O, halogen, CN, OH, NH2, -C(=O)NH2, NHC 1-4 Alkyl, N(C) 1-4 Alkyl)2, C 1-4 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl group, -OC 1-4 Alkyl, -SC 1-4 Alkyl, -OC 3-6 Carbocyclic rings, -O-3 to 7-membered heterocycles, C 3-6 Carbocyclic rings, 3 to 7-membered heterocycles, wherein the alkyl, alkenyl, alkynyl, carbocyclic, or heterocycle is optionally selected from 1 to 4 elements selected from deuterium, halogens, CN, OH, NH2, C. 1-4 Alkyl, C 1-4 The alkoxy group is replaced by a substituent. According to claim 2, the compound or its stereoisomers, tautomers, racemates, or pharmaceutically acceptable salts thereof, R is selected from -C(=O)R q1 -P(=O)R q1 R q2 -CH2-OP(=O)R q1 R q2 -C(O)O-CH2-OP(=O)R q1 R q2 -S(=O)2C 1-6 Alkyl group, -S(=O)2C 3-6 cycloalkyl, C 3-6 Carbocyclic, 4- to 7-membered heterocyclic, wherein the CH2, cycloalkyl, carbocyclic or heterocyclic is optionally surrounded by 1 to 4 R... k replace; R q1 R q2 Each is independently selected from H, OH, or arbitrarily selected by 1 to 4 R. k The substitution is made with one of the following groups: methyl, ethyl, propyl, butyl, isopropyl, tert-butyl, methoxy, ethoxy, isopropyloxy, tert-butyloxy, cyclopropyl, cyclobutyl, cyclopentyl, oxecyclobutyl, oxecyclopentyl, oxecyclohexyl, azircyclobutyl, azircyclopentyl, azircyclohexyl; R q3 R q4 Each is independently selected from H or arbitrarily selected by 1 to 4 Rs. k The substitution is performed using one of the following groups: methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, -CH2NH2, -CH2CH2NH2, -CH2CH2CH2NH2, -CH2CH2CH2CH2NH2, -CH2NHC(=NH)NH2, -CH2CH2NHC(=NH)NH2, -CH2CH2CH2NHC(=NH)NH2, -CH2CH2CH2NHC(=O)NH2, -CH2OH, -CH2SH, -CH2CH2SH, -CH2CO2H, -CH2 CH2CO2H, -CH2C(=O)NH2, -CH2CH2C(=O)NH2, -CH2CH2CH2C(=O)NH2, -CH2CH2CH2CH2C(=O)NH2, -CH2-phenyl, -CH2-indole, -CH2-5 to 10-membered heteroaryl, -CH2-C 3-6 cycloalkyl, C 3- 6-membered cycloalkyl, 4- to 7-membered heterocyclic alkyl, or -CH2-4- to 7-membered heterocyclic alkyl; R 1 R 2 R 3 R 4 R 5 Each of the following groups is independently selected from H, F, Cl, Br, cyano, methyl, ethyl, methoxy, ethoxy, methylthio, cyclopropyl, cyclobutyl, vinyl, and ethynyl, wherein the methyl, ethyl, methoxy, ethoxy, methylthio, cyclopropyl, cyclobutyl, vinyl, and ethynyl groups are optionally prefixed with 1 to 4 R groups. k replace; As an option, R 1 R 4 Direct connection forms C 3-6 Carbon ring, wherein the carbon ring is optionally divided by 1 to 4 R k replace; B is selected from Or phenyl, wherein the B is optionally surrounded by 1 to 4 R B replace; R q R B Each of the following groups is independently selected from H, deuterium, =O, F, Cl, Br, cyano, methyl, ethyl, isopropyl, methoxy, ethoxy, isopropyloxy, methylthio, cyclopropyl, cyclopropyl-O-, cyclobutyl, vinyl, ethynyl, -P(=O)(CH3)2, -P(=O)(CH2CH3)2, -P(=O)(CH3)(cyclopropyl), wherein the methyl, ethyl, isopropyl, methoxy, ethoxy, isopropyloxy, methylthio, cyclopropyl, cyclobutyl, vinyl, and ethynyl groups are optionally prefixed with 1 to 4 R groups. k replace; R k Each is independently selected from deuterium, =O, F, Cl, Br, I, CN, OH, NH2, -C(=O)NH2, NH(CH3), NH(CH2CH3), N(CH3)2, N(CH2CH3)2, methyl, ethyl, vinyl, ethynyl, methoxy, ethoxy, methylthio, -O-cyclopropyl, -NH-cyclopropyl, -CH2-cyclopropyl, -CH2-cyclobutyl, -CH2-cyclopentyl -CH2-cyclohexyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, wherein the methyl, ethyl, vinyl, ethynyl, methoxy, ethoxy, methylthio, -O-cyclopropyl, -NH-cyclopropyl, -CH2-cyclopropyl, -CH2-cyclobutyl, -CH2-cyclopentyl, -CH2-cyclohexyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, optionally selected from 1 to 4 elements selected from deuterium, halogen, CN, OH, NH2, C 1-4 Alkyl, C 1-4 The alkoxy group is replaced by a substituent. According to claim 3, the compound or its stereoisomers, tautomers, racemates, or pharmaceutically acceptable salts thereof, wherein, R is selected from -C(=O)R q1 -P(=O)R q1 R q2 -CH2-OP(=O)R q1 R q2 -C(O)O-CH2-OP(=O)R q1 R q2 -S(=O)2R q1 The CH2 is optionally converted by 1 to 2 R k replace; R q3 R q4 Each is independently selected from H or arbitrarily selected by 1 to 4 Rs. k The substitution is made with one of the following groups: methyl, ethyl, propyl, butyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, -CH2NH2, -CH2CH2NH2, -CH2CH2CH2NH2, -CH2CH2CH2CH2NH2, -CH2NHC(=NH)NH2, -CH2CH2NHC(=NH)NH2, -CH2CH2CH2NHC(=NH)NH2, -CH2CH2CH2NHC(=O)NH2, -CH2OH, -CH2SH, -CH2CH2SH, -CH2CO2H, -CH2 CH2CO2H, -CH2-phenyl, -CH2-imidazolyl, -CH2-indolyl, cyclopropyl, cyclobutyl, cyclopentyl, oxecyclobutyl, oxecyclopentyl, oxecyclohexyl, azircyclobutyl, azircyclopentyl, azircyclohexyl, -CH2-cyclopropyl, -CH2-cyclobutyl, -CH2-cyclopentyl, -CH2-oxecyclobutyl, -CH2-oxecyclopentyl, -CH2-azircyclobutyl, -CH2-azircyclopentyl, -CH2-azircyclohexyl; As an option, R q3 R q4 Directly linked to form cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, oxacyclobutyl, oxacyclopentyl, oxacyclohexyl, aziridine, aziridinepentyl, and aziridinehexyl, wherein the cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, oxacyclobutyl, oxacyclopentyl, oxacyclohexyl, aziridine, aziridinepentyl, and aziridinehexyl are optionally separated by 1 to 4 R... k replace; Q1 is selected from Q1 is arbitrarily selected by 1 to 4 Rs q replace; R 1 R 2 R 3 R 4 R 5 Each is independently selected from H, methyl, ethyl, CH2F, CHF2, CF3; R q R B Each group is independently selected from H, =O, F, Cl, Br, cyano, CH2F, CHF2, CF3, -OCH2F, -OCHF2, -OCF3, -OCD3, methyl, -S-methyl, -S-CF3, ethyl, isopropyl, ethynyl, methoxy, ethoxy, isopropyloxy, propyloxy, cyclopropyl, -O-cyclopropyl, -P(=O)(CH3)2, -P(=O)(CH2CH3)2, -P(=O)(CH3)(cyclopropyl), wherein the methyl, ethyl, isopropyl, ethynyl, methoxy, ethoxy, isopropyloxy, propyloxy, and cyclopropyl groups are optionally prefixed with 1 to 4 R groups. k replace; R k Each is independently selected from deuterium, F, Cl, Br, I, CN, OH, NH2, -C(=O)NH2, -CH2OH, methyl, ethyl, vinyl, ethynyl, methoxy, ethoxy, methylthio, -O-cyclopropyl, -NH-cyclopropyl, -CH2-cyclopropyl, -CH2-cyclobutyl, -CH2-cyclopentyl, -CH2-cyclohexyl, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl. According to claim 4, the compound or its stereoisomers, tautomers, racemates, or pharmaceutically acceptable salts thereof, wherein, Q1 is selected from Q1 is arbitrarily selected by 1 to 3 Rs q replace; B is selected from According to claim 5, the compound or its stereoisomers, tautomers, racemates, or pharmaceutically acceptable salts thereof, wherein, X is selected from O and S; R is selected from Preferably, R is selected from B is selected from The compound according to claim 1, or its stereoisomers, tautomers, racemates, or pharmaceutically acceptable salts, wherein the compound represented by formula (I) is selected from formula (Ib). R 4 Selected from H or C 1-4 Alkyl, preferably H or methyl; R b Each is independently selected from deuterium, halogens, CN, and C. 1-6 Alkyl, -OC 1-6 Alkyl group, wherein the alkyl group is optionally substituted with 1 to 4 substituents selected from deuterium, halogen, cyclopropyl, preferably F, Cl, Br, methyl, ethyl, methoxy, ethoxy, CH2F, CHF2, CF3, -OCH2F, -OCHF2, -OCF3, -OCD3, -OCH2D, -OCHD2; R q Each is independently selected from deuterium, halogens, CN, and C. 1-6 Alkyl, -OC 1-6 Alkyl group, wherein the alkyl group is optionally substituted with 1 to 4 substituents selected from deuterium, halogen, cyclopropyl, preferably F, Cl, or Br; R is selected from -P(=O)(OH)2; n1 and n2 are selected from 0, 1, 2, and 3. The compound according to claim 1 or its stereoisomers, tautomers, racemates, or pharmaceutically acceptable salts, wherein the compound is selected from one of the structures shown in Table E. The compound or its stereoisomers, tautomers, racemates, or pharmaceutically acceptable salts according to any one of claims 1-8, wherein the salt is selected from acid or base molecules, and the base molecule is selected from at least one of lithium salts, sodium salts, potassium salts, calcium salts, magnesium salts, aluminum salts, iron salts, zinc salts, ammonium salts, lysine salts, arginine salts, L-arginine salts, histidine salts, L-histidine salts, meglumine salts, dimethylglucosamine salts, ethylglucosamine salts, dicyclohexylamine salts, 1,6-hexanediamine salts, glucosamine salts, triethanolamine salts, sarcosine salts, serine salts, trihydroxymethylaminomethane salts, aminopropylene glycol salts, 1-amino-2,3,4-butanetriol salts, L-lysine salts, ornithine salts, preferably meglumine salts; the acid molecule is selected from at least one of hydrochloric acid, sulfuric acid, hydrobromic acid, phosphoric acid, citric acid, oxalic acid, maleic acid, fumaric acid, L-tartaric acid, and p-toluenesulfonic acid. According to claim 9, the compound or its stereoisomers, tautomers, racemates, or pharmaceutically acceptable salts thereof, wherein the stoichiometric ratio of the compound to the acid or base molecule is 1:2 to 1:1 or 1:1 to 2:1, preferably 1:2, 1:1, or 2:

1. The compound according to claim 9, or its stereoisomers, tautomers, racemates, or pharmaceutically acceptable salts, is characterized in that... The salt mentioned is A pharmaceutical composition comprising the compound of any one of claims 1-10 or its stereoisomers, tautomers, racemates, pharmaceutically acceptable salts, and a pharmaceutically acceptable carrier, preferably comprising 1-1500 mg of the compound of any one of claims 1-10 or its stereoisomers, tautomers, racemates, or pharmaceutically acceptable salts. The use of the compound of any one of claims 1-11, or its stereoisomers, tautomers, racemates, pharmaceutically acceptable salts, or the pharmaceutical composition of claim 12 in the preparation of a medicament for treating and / or alleviating pain or pain-related diseases, multiple sclerosis, Sharma-Tutankhamun syndrome, incontinence, pathological cough, or arrhythmia; preferably, the pain is selected from chronic pain, acute pain, inflammatory pain, cancer pain, postoperative pain, neuropathic pain, musculoskeletal pain, primary pain, intestinal pain, and idiopathic pain; the postoperative pain is preferably selected from pain following bunion excision, hernia repair, and abdominoplasty. A method for treating or alleviating a disease in a mammal, the method comprising administering to a subject a therapeutically effective amount of the compound of any one of claims 1-11 or its stereoisomers, tautomers, racemates, pharmaceutically acceptable salts, or the pharmaceutical composition of claim 12, wherein the therapeutically effective amount is preferably 1-1500 mg, and the disease is preferably pain, pain-related diseases, multiple sclerosis, Sharma-Tutankhamun syndrome, incontinence, pathological cough, or arrhythmia.

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