Transient receptor potential cation channel TRPV3 inhibitor and application thereof

Abstract

The invention belongs to the technical field of medicines, and particularly relates to a transient receptor potential cation channel TRPV3 inhibitor and application thereof. The compound provided by the invention can be used as a TRPV3 inhibitor, can inhibit abnormal increase of calcium ion influx caused by TRPV3 activation, can effectively treat and prevent TRPV3-related diseases, and provides a new idea for diseases such as pruritus, alopecia, atopic dermatitis, psoriasis, ulcerative colitis and the like. The compound provided by the invention has better dissolvability on the premise of equivalent inhibition rate to calcium ion internal flow; meanwhile, the compound disclosed by the invention has a proliferation inhibition effect on human hepatic stellate cells LX-2 and has an anti-fibrosis effect.

Description

Technical Field

[0001] The present invention belongs to the field of medical technology, and specifically relates to a quinolinone compound with TRPV3 inhibitory effect and a pharmaceutical composition thereof, and further relates to a preparation method and pharmaceutical use thereof. Background Art

[0002] TRPV3 is a non-selective cation channel with relatively high permeability to calcium ions. It was first cloned in 2002 and shows 30%-40% sequence homology with other TRPV channels. TRPV3 channels are homotetramers formed by six transmembrane subunits, containing multiple repeats of the ankyrin domain ARD, amino-terminal and carboxyl-terminal, and a pore-forming loop between the domains.

[0003] As a member of the temperature-sensitive TRP family, the calcium-permeable nonselective cation channel TRPV3 is abundantly expressed in skin keratinocytes. Weak acids can diffuse across the cell membrane in a protonated form, subsequently releasing free protons, leading to intracellular acidification and activation of TRPV3. Protons can also pass through activated TRPV3 or other proton-permeable channels, leading to intracellular acidification and activation of TRPV3. TRPV3 activation mediates calcium influx and induces calcium ion cytoplasmic overload, leading to keratinization and cell death or skin disease with severe mast cell infiltration and histamine release in the upper dermis; in addition, TRPV3-mediated nitric oxide release in keratinocytes promotes wound healing and causes pain. The recent discovery of gain-of-function mutations in human TRPV3 from patients with Olmsted syndrome, which is characterized by severe pruritus and palmoplantar and perioral keratosis, revealed its key role in chronic skin diseases. In mice, loss-of-function mutations in transforming growth factor α (TGF-α) and epidermal growth factor receptor (EGFR) genes cause a wavy hair phenotype, whereas upregulation of TGF-α / EGFR signaling results in a hairless phenotype. Furthermore, small interfering RNA-mediated silencing of TRPV3 effectively abolished TRPV3 agonist-induced cellular effects, including functional currents, reduced proliferation, elevated intracellular calcium concentrations, and apoptosis.

[0004] Currently, many scientists have begun to focus on the discovery of TRPV3 inhibitors. TRPV3 forms a signaling complex with TGF-a / EGFR. 2+Influx leads to the release / shedding of TGF-α, which in turn activates the signaling pathway of EGFRs to promote keratinocyte proliferation. The pathogenesis of psoriasis is related to the excessive proliferation of immune cells and skin keratinocytes and the massive release of inflammatory factors after the immune response occurs. At the same time, the activation of EGFR leads to an increase in the activity of TRPV3 channels, thereby stimulating the release of TGF-a. In short, selective targeted inhibition of TRPV3 may be an effective method for the treatment of various skin diseases such as skin keratinization, hair loss, itching, psoriasis, etc.

[0005] Glenmark has applied for patents US20100292254, US20090286811, and US20100311778 for a series of TRPV3 antagonists. In 2010, Glenmark signed an out-licensing agreement with Sanofi-Aventis and subsequently promoted its lead molecule (GRC15300, unknown structure) to the clinic for the treatment of osteoarthritis and neuropathic pain. In 2012, GRC15300 entered Phase II trials for the treatment of neuropathic pain; however, by the end of 2013, these trials were announced to be stopped. The TRPV3 inhibitor small molecule structure is as follows:

[0006]

[0007] A recently published article (Wang, Y., Tan, L., Jiao, K., et al. Scutellareinattenuates atopic dermatitis by selectively inhibiting transient receptor potential vanilloid 3 channels. British journal of pharmacology, 179 (20), 4792–4808.) describes scutellarin as an allosteric negative regulator of TRPV3 channels with an apparent affinity of 1.18 μM. It can weaken the endogenous TRPV3 activity in human keratinocytes and inhibit the proliferation and pro-inflammatory response induced by carvacrol. The scutellarin structural formula is as follows:

[0008] Summary of the invention

[0009] The technical problem to be solved by the present invention is to address the deficiencies of the prior art and provide a quinolinone compound as a TRPV3 inhibitor and a pharmaceutical composition thereof, which can be used to treat various inflammatory skin disease-related diseases, such as psoriasis.

[0010] In order to solve the above technical problems, the technical solution adopted by the present invention is as follows:

[0011] The present invention discloses a compound represented by formula I or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs;

[0012]

[0013] in,

[0014] R1 is independently selected from hydrogen, deuterium, substituted or unsubstituted amino, hydroxyl, thiol, cyano, nitro, substituted or unsubstituted C 1-6 Alkyl, substituted or unsubstituted C 1-6 Alkoxy, substituted or unsubstituted C 1-6 Haloalkyl, substituted or unsubstituted C 1-6 Haloalkoxy, substituted or unsubstituted C 1-6 Alkylamino, carboxyl, substituted or unsubstituted C 1-6 Alkanoyl, substituted or unsubstituted C 1-6 alkylsulfonyl, substituted or unsubstituted aminoacyl, or substituted or unsubstituted aminosulfonyl; wherein the substitution is selected from hydrogen, deuterium, halogen, hydroxyl or C 1-6 Alkyl substitution;

[0015] R2 is independently selected from hydrogen, deuterium, substituted or unsubstituted amino, hydroxyl, thiol, cyano, nitro, halogen, substituted or unsubstituted C 1-6 Alkyl, substituted or unsubstituted C 1-6 Alkoxy, substituted or unsubstituted C 1-6 Haloalkyl, substituted or unsubstituted C 1-6 Haloalkoxy, substituted or unsubstituted C 1-6 Alkylamino, carboxyl, substituted or unsubstituted C 1-6 Alkanoyl, substituted or unsubstituted C 1-6 alkylsulfonyl, substituted or unsubstituted aminoacyl, or substituted or unsubstituted aminosulfonyl; wherein the substitution is selected from hydrogen, deuterium, halogen, hydroxyl or C 1-6 Alkyl substitution;

[0016] n is an integer selected from 1 to 4;

[0017] m is an integer selected from 0-3;

[0018] Virtual bonds are represented as non-existent, single bonds, or double bonds, and virtual bonds are not always double bonds;

[0019] When the virtual bond is represented as absent, X is selected from -NR3R4, -CR5R6R7, -OR8 or -SR9;

[0020] When the virtual bond connected to X is represented as a single bond, X is selected from -NR3-, -CR5R6-, -O- or -S-;

[0021] R3 is independently selected from hydrogen, deuterium, substituted or unsubstituted amino, hydroxyl, thiol, cyano, nitro, substituted or unsubstituted C 1-6 Alkyl, substituted or unsubstituted C 1-6 Alkoxy, substituted or unsubstituted C 1-6 Haloalkyl, substituted or unsubstituted C 1-6 Haloalkoxy, substituted or unsubstituted C 1-6 Alkylamino, carboxyl, substituted or unsubstituted C 1-6 Alkanoyl, substituted or unsubstituted C 1-6 alkylsulfonyl, substituted or unsubstituted aminoacyl, or substituted or unsubstituted aminosulfonyl; wherein the substitution is selected from hydrogen, deuterium, halogen, hydroxyl or C 1-6 Alkyl substitution;

[0022] R4 is independently selected from hydrogen, deuterium, substituted or unsubstituted amino, hydroxyl, thiol, cyano, nitro, substituted or unsubstituted C 1-6 Alkyl, substituted or unsubstituted C 1-6 Alkoxy, substituted or unsubstituted C 1-6 Haloalkyl, substituted or unsubstituted C 1-6 Haloalkoxy, substituted or unsubstituted C 1-6 Alkylamino, carboxyl, substituted or unsubstituted C 1-6 Alkanoyl, substituted or unsubstituted benzoyl, substituted or unsubstituted C 1-6 alkylsulfonyl, substituted or unsubstituted aminoacyl, or substituted or unsubstituted aminosulfonyl; wherein the substitution is selected from hydrogen, deuterium, halogen, hydroxyl and C 1-6 The alkyl group may be substituted by one or more identical or different groups;

[0023] R5, R6, R7, R8, and R9 are independently selected from hydrogen, deuterium, substituted or unsubstituted amino, hydroxyl, thiol, cyano, nitro, substituted or unsubstituted C 1-6 Alkyl, substituted or unsubstituted C 1-6 Alkoxy, substituted or unsubstituted C 1-6 Haloalkyl, substituted or unsubstituted C 1-6 Haloalkoxy, substituted or unsubstituted C 1-6 Alkylamino, carboxyl, substituted or unsubstituted C 1-6 Alkanoyl, substituted or unsubstituted C 1-6alkylsulfonyl, substituted or unsubstituted aminoacyl, substituted or unsubstituted aminosulfonyl, substituted or unsubstituted C 3-8 Cycloalkyl, substituted or unsubstituted C 6-10 Aryl, substituted or unsubstituted heterocyclic group consisting of 3 to 8 atoms or substituted or unsubstituted heteroaryl group consisting of 5 to 10 atoms.

[0024] In some embodiments, preferably, R1 is independently selected from hydrogen, deuterium, substituted or unsubstituted amino, hydroxyl, thiol, cyano, nitro, substituted or unsubstituted C 1-3 Alkyl, substituted or unsubstituted C 1-3 Alkoxy, substituted or unsubstituted C 1-3 Haloalkyl, substituted or unsubstituted C 1-3 Haloalkoxy, substituted or unsubstituted C 1-3 Alkylamino, carboxyl, substituted or unsubstituted C 1-3 Alkanoyl, substituted or unsubstituted C 1-3 alkylsulfonyl, substituted or unsubstituted aminoacyl, or substituted or unsubstituted aminosulfonyl; wherein the substitution is selected from hydrogen, deuterium, halogen, hydroxyl or C 1-3 Alkyl substitution;

[0025] R2 is independently selected from hydrogen, deuterium, substituted or unsubstituted amino, hydroxyl, thiol, cyano, nitro, halogen, substituted or unsubstituted C 1-3 Alkyl, substituted or unsubstituted C 1-3 Alkoxy, substituted or unsubstituted C 1-3 Haloalkyl, substituted or unsubstituted C 1-3 Haloalkoxy, substituted or unsubstituted C 1-3 Alkylamino, carboxyl, substituted or unsubstituted C 1-3 Alkanoyl, substituted or unsubstituted C 1-3 alkylsulfonyl, substituted or unsubstituted aminoacyl, or substituted or unsubstituted aminosulfonyl; wherein the substitution is selected from hydrogen, deuterium, halogen, hydroxyl or C 1-3 Alkyl substitution;

[0026] n is preferably an integer selected from 1 to 3;

[0027] m is preferably selected from an integer of 0-2;

[0028] Virtual bonds are represented as non-existent, single bonds, or double bonds, and virtual bonds are not always double bonds;

[0029] When the virtual bond is indicated as absent, X is preferably selected from -NR3R4 or -OR8;

[0030] When the virtual bond connected to X is represented as a single bond, X is preferably selected from -NR3- or -O-;

[0031] R3 is independently selected from hydrogen, deuterium, substituted or unsubstituted amino, hydroxyl, thiol, cyano, nitro, substituted or unsubstituted C 1-3 Alkyl, substituted or unsubstituted C 1-3 Alkoxy, substituted or unsubstituted C 1-3 Haloalkyl, substituted or unsubstituted C 1-3 Haloalkoxy, substituted or unsubstituted C 1-3 Alkylamino, carboxyl, substituted or unsubstituted C 1-3 Alkanoyl, substituted or unsubstituted C 1-3 alkylsulfonyl, substituted or unsubstituted aminoacyl, or substituted or unsubstituted aminosulfonyl; wherein the substitution is selected from hydrogen, deuterium, halogen, hydroxyl or C 1-3 Alkyl substitution;

[0032] R4 is independently selected from hydrogen, deuterium, substituted or unsubstituted amino, hydroxyl, thiol, cyano, nitro, substituted or unsubstituted C 1-3 Alkyl, substituted or unsubstituted C 1-3 Alkoxy, substituted or unsubstituted C 1-3 Haloalkyl, substituted or unsubstituted C 1-3 Haloalkoxy, substituted or unsubstituted C 1-3 Alkylamino, carboxyl, substituted or unsubstituted C 1-3 Alkanoyl, substituted or unsubstituted benzoyl, substituted or unsubstituted C 1-3 alkylsulfonyl, substituted or unsubstituted aminoacyl, or substituted or unsubstituted aminosulfonyl; wherein the substitution is selected from hydrogen, deuterium, halogen, hydroxyl and C 1-3 The alkyl group may be substituted by one or more identical or different groups;

[0033] R5, R6, R7, R8, and R9 are independently selected from hydrogen, deuterium, substituted or unsubstituted amino, hydroxyl, thiol, cyano, nitro, substituted or unsubstituted C 1-3 Alkyl, substituted or unsubstituted C 1-3 Alkoxy, substituted or unsubstituted C 1-3 Haloalkyl, substituted or unsubstituted C 1-3 Haloalkoxy, substituted or unsubstituted C 1-3 Alkylamino, carboxyl, substituted or unsubstituted C 1-3 Alkanoyl, substituted or unsubstituted C 1-3 alkylsulfonyl, substituted or unsubstituted aminoacyl, substituted or unsubstituted aminosulfonyl, substituted or unsubstituted C 3-5Cycloalkyl, substituted or unsubstituted C 6-8 Aryl, substituted or unsubstituted heterocyclic group consisting of 3 to 8 atoms or substituted or unsubstituted heteroaryl group consisting of 5 to 10 atoms.

[0034] In some embodiments, it is further preferred that the compound represented by the above formula I or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs,

[0035] When the virtual bond connected to X is represented as a single bond, and the other virtual bond is represented as a double bond, the compound of formula I is represented by formula I-1:

[0036]

[0037] in,

[0038] R1 is independently selected from hydrogen, deuterium, hydroxyl or substituted or unsubstituted C 1-3 wherein the substitution is selected from hydrogen, deuterium, hydroxyl or C 1-3 Alkyl substitution;

[0039] R2 is independently selected from substituted or unsubstituted C 1-3 Alkyl, hydrogen, deuterium, hydroxyl or halogen; the substitution is selected from hydrogen, deuterium, halogen, hydroxyl or C 1-3 Alkyl substitution;

[0040] n is further preferably selected from 2 or 3;

[0041] m is further preferably selected from 1 or 2;

[0042] X is preferably selected from -NR3- or -O-;

[0043] R3 is independently selected from hydrogen or deuterium.

[0044] Specifically, more preferably, R1 is independently selected from hydrogen, deuterium, hydroxyl or substituted or unsubstituted ethyl; wherein the substitution is selected from substitution with methyl;

[0045] R2 is independently selected from hydrogen, deuterium, hydroxy, isopropyl, fluorine or chlorine;

[0046] R3 is independently selected from hydrogen or deuterium.

[0047] In some embodiments, it is further preferred that the compound represented by the above formula I or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs,

[0048] When X is selected from -O-, n is selected from 3, and m is selected from 1, the compound of formula I is as shown in formula I-1-1:

[0049]

[0050] R 10 independently selected from hydrogen, deuterium, hydroxyl or substituted or unsubstituted ethyl; wherein the substitution is selected from hydrogen, deuterium, hydroxyl or C 1-3 Alkyl substitution;

[0051] R 11 , R 12 independently selected from hydrogen, deuterium, hydroxyl;

[0052] R2 is independently selected from hydrogen, deuterium, and hydroxyl.

[0053] Specifically, more preferably, R 10 are independently selected from hydrogen, deuterium, hydroxyl or substituted or unsubstituted ethyl; wherein the substitution is selected from C 1-3 Alkyl substitution.

[0054] In some embodiments, it is further preferred that the compound represented by the above formula I or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs,

[0055] When X is selected from -NR3-, n is selected from 2, and m is selected from 1, the compound of formula I is shown in formula I-1-2:

[0056]

[0057] R1 is independently selected from hydroxyl;

[0058] R3 is independently selected from hydrogen or deuterium;

[0059] R 13 independently selected from substituted or unsubstituted ethyl; wherein the substitution is selected from hydrogen, deuterium, hydroxyl or C 1-3 Alkyl substitution.

[0060] Specifically, more preferably, R 13 independently selected from substituted or unsubstituted ethyl; wherein the substitution is selected from C 1-3 Alkyl substitution.

[0061] In some embodiments, it is further preferred that the compound represented by the above formula I or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs,

[0062] When X is selected from -NR3-, n is selected from 2, and m is selected from 2, the compound of formula I is shown in formula I-1-3:

[0063]

[0064] R1 is independently selected from hydroxyl;

[0065] R3 is independently selected from hydrogen or deuterium;

[0066] R 14 , R 15 Independently selected from hydroxy or halogen.

[0067] Specifically, more preferably, R 14 , R 15 are independently selected from hydroxy, fluoro or chloro.

[0068] In some embodiments, it is further preferred that the compound represented by the above formula I or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs,

[0069] When all virtual bonds are indicated as non-existent, the compound of formula I is as shown in formula I-2:

[0070]

[0071] R1 is independently selected from hydrogen, deuterium or hydroxyl;

[0072] n is further preferably selected from 2;

[0073] X is further preferably selected from -NR3R4;

[0074] R3 is independently selected from hydrogen, deuterium or C 1-3 alkyl;

[0075] R4 is independently selected from hydrogen, deuterium or substituted or unsubstituted benzoyl; wherein the substitution is selected from substitution by one or more identical or different groups selected from hydrogen, deuterium, halogen and hydroxyl.

[0076] Specifically, more preferably, R1 is independently selected from hydrogen, deuterium or hydroxyl;

[0077] R3 is independently selected from hydrogen or deuterium;

[0078] R4 is independently selected from substituted or unsubstituted benzoyl; wherein the substitution is selected from substitution by one or more identical or different groups selected from hydrogen, deuterium, fluorine, chlorine and hydroxyl.

[0079] In some embodiments, most preferably, the compound represented by the above formula I or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs is selected from any of the following compounds:

[0080]

[0081] The compound represented by the above formula I or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs are prepared by the following method, selecting one of the following synthetic routes:

[0082] Synthetic route 1:

[0083]

[0084] wherein R2 is selected from substituted or unsubstituted ethyl, hydroxyl or halogen; wherein the substitution is selected from hydrogen, deuterium, hydroxyl or C 1-3 Alkyl substituted; m is selected from 1 or 2;

[0085] or,

[0086] Synthetic route 2:

[0087]

[0088] Among them, R 16 is selected from hydrogen, deuterium, halogen or hydroxyl; z is selected from 2;

[0089] or,

[0090] Synthetic route 3:

[0091]

[0092] Furthermore, the present invention discloses a method for preparing the compound represented by the above formula I or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs, and selects one of the following synthetic routes:

[0094] Synthetic route 1:

[0095] 2-amino-4,5-dimethoxyacetophenone and compound a-1 undergo a first acylation reaction under the action of a first base to obtain intermediate b-1; intermediate b-1 undergoes a first intramolecular cyclization reaction under the action of a second base to obtain intermediate c-1; intermediate c-1 undergoes a first demethylation reaction under the action of a first Lewis acid to obtain compound IA;

[0096]

[0097] wherein R2 is selected from substituted or unsubstituted ethyl, hydroxyl or halogen; wherein the substitution is selected from hydrogen, deuterium, hydroxyl or C 1-3 Alkyl substituted; m is selected from 1 or 2;

[0098] or,

[0099] Synthetic route 2:

[0100] 2-amino-4,5-dimethoxyacetophenone and compound a-2 undergo a second acylation reaction under the action of a third base to obtain intermediate b-2; intermediate b-2 undergoes a second demethylation reaction under the action of a second Lewis acid to obtain compound IB;

[0101]

[0102] Among them, R 16 is selected from hydrogen, deuterium, halogen or hydroxyl; z is selected from 2;

[0103] or,

[0104] Synthetic route 3:

[0105] 3,5-dimethoxyphenol and 2-chloropropane undergo a first substitution reaction under the action of a first catalyst to obtain compound a-3; compound a-3 and acetyl chloride undergo a second substitution reaction under the action of a second catalyst to obtain compound b-3; compound b-3 and 4-methoxybenzoyl chloride undergo an esterification rearrangement reaction under the action of a fourth base to obtain compound c-3; compound c-3 undergoes a cyclization reaction under the action of an inorganic acid to obtain compound d-3; compound d-3 undergoes a third demethylation reaction under the action of a third Lewis acid to obtain compound 1;

[0106]

[0107] Specifically, in the synthetic route 1, the first base is triethylamine, ethylenediamine or n-propylamine, preferably triethylamine; the molar ratio of the 2-amino-4,5-dimethoxyacetophenone to the compound a-1 and the first base is 1:0.9-4.5:2-70:, preferably 1:1.2-3:2.4-14, and more preferably 1:1.5:2.8; the solvent used in the first acylation reaction is tetrahydrofuran, methyltetrahydrofuran or dimethyltetrahydrofuran, preferably tetrahydrofuran; the first Acylation reaction, the reaction temperature is 45-85°C, preferably 55-75°C, more preferably 65°C, the reaction time is 2.4-12h, preferably 3.2-8h, more preferably 4h; the second base is sodium hydroxide, potassium hydroxide or calcium hydroxide, preferably sodium hydroxide; the molar ratio of the intermediate b-1 to the second base is 1:4-26, preferably 1:5-13, more preferably 1:6-7.2, and more preferably 1:6.6; the first intramolecular cyclization reaction The solvent used in the reaction is toluene, xylene or 1,4-dioxane, preferably 1,4-dioxane; the reaction temperature of the first intramolecular cyclization reaction is 90-130°C, preferably 100-120°C, more preferably 110°C, and the reaction time is 2.4-12h, preferably 3.2-8h, and more preferably 4h; the first Lewis acid is boron tribromide, boron trichloride or aluminum trichloride, preferably boron tribromide; the molar ratio of the intermediate c-1 to the first Lewis acid is 1:4-26, preferably 1:5-13, more preferably 1:6-6.5, and more preferably 1:6.25; the first Lewis acid exists in the form of a solution, the solvent is dichloromethane, and the concentration of the first Lewis acid in the solution is 1.2-6mol / L, preferably 1.6-4mol / L, and more preferably 2mol / L; the reaction temperature of the first demethylation reaction is room temperature, and the reaction time is 2.4-12h, preferably 3.2-8h, and more preferably 4h.

[0108] Specifically, in synthetic route 1, the first acylation reaction is preferably carried out under the protection of an inert gas; the first intramolecular cyclization reaction is preferably carried out under the protection of an inert gas; the first demethylation reaction is preferably carried out under the protection of an inert gas; wherein the inert gas is preferably nitrogen.

[0109] Specifically, in synthetic route 2, the third base is triethylamine, ethylenediamine or n-propylamine, preferably triethylamine; the molar ratio of the 2-amino-4,5-dimethoxyacetophenone to the compound a-2 and the third base is 1:0.9-4.5:2-70:, preferably 1:1.2-3:2.4-14, and more preferably 1:1.5:2.8; the solvent used in the second acylation reaction is tetrahydrofuran, methyltetrahydrofuran or dimethyltetrahydrofuran, preferably tetrahydrofuran; the second acylation reaction, the reaction temperature is 45-85°C, preferably 55-75°C, and more preferably 65°C, the reaction time is 2.4-12h, preferably 3.2-8h, and further step is preferably 4h; the second Lewis acid is boron tribromide, boron trichloride or aluminum trichloride, preferably boron tribromide; the molar ratio of the intermediate b-2 to the second Lewis acid is 1:4~26, preferably 1:5~13, further preferably 1:6~6.5, and further preferably 1:6.25; the second Lewis acid exists in the form of a solution, the solvent is dichloromethane, and the concentration of the second Lewis acid in the solution is 1.2~6mol / L, preferably 1.6~4mol / L, and further preferably 2mol / L; the second demethylation reaction, the reaction temperature is room temperature, the reaction time is 2.4~12h, preferably 3.2~8h, and further preferably 4h.

[0110] Specifically, in synthetic route 2, the second acylation reaction is preferably carried out under the protection of an inert gas; the second demethylation reaction is preferably carried out under the protection of an inert gas; wherein the inert gas is preferably nitrogen.

[0111] Specifically, in synthetic route 3, the first catalyst is boron tribromide, boron trichloride or aluminum trichloride, preferably aluminum trichloride; the molar ratio of the 3,5-dimethoxyphenol to 2-chloropropane and the first catalyst is 1:0.8-2.4:2.1-8.1, preferably 1:1-1.8:2.4-5.4, and more preferably 1:1.2:2.7; the solvent used in the first substitution reaction is dichloromethane or chloroform, preferably chloroform; the first substitution reaction, the reaction temperature is room temperature, the reaction time is 2.4-12h, preferably 3.2-8h, and more preferably 4h; the second catalyst is boron tribromide, boron trichloride or aluminum trichloride, preferably is aluminum chloride; the molar ratio of the compound a-3 to acetyl chloride and the second catalyst is 1:0.8-2.4:1.2-6, preferably 1:1-1.8:1.6-4, and more preferably 1:1.2:2; the solvent used in the second substitution reaction is dichloromethane or chloroform, preferably chloroform; the reaction temperature of the second substitution reaction is room temperature, and the reaction time is 2.4-12h, preferably 3.2-8h, and more preferably 4h; the fourth base is sodium hydroxide, potassium hydroxide or calcium hydroxide, and is preferably sodium hydroxide; the molar ratio of the compound b-3 to 4-methoxybenzoyl chloride and the fourth base is 1:0.6-3:1.6-38.4, preferably The molar ratio of compound c-3 to inorganic acid is 1:0.24-4.32, preferably 1:0.28-1.44, and more preferably 1:0.37; the solvent used in the cyclization reaction is acetic acid, formic acid or oxalic acid, preferably acetic acid; the reaction temperature of the cyclization reaction is room temperature, and the reaction time is 1.8-12h, preferably 2. 5~6h, more preferably 3h; the third Lewis acid is boron tribromide, boron trichloride or aluminum trichloride, preferably boron tribromide; the molar ratio of the compound d-3 to the third Lewis acid is 1:9.6~230.4, preferably 1:12~57.6, more preferably 1:14.2; the third Lewis acid exists in the form of a solution, the solvent is dichloromethane, and the concentration of the third Lewis acid in the solution is 1.2~6mol / L, preferably 1.6~4mol / L, more preferably 2mol / L; the third demethylation reaction, the reaction temperature is room temperature, the reaction time is 2.4~12h, preferably 3.2~8h, more preferably 4h.

[0112] Specifically, in the synthetic route 3, the esterification rearrangement reaction is preferably carried out under the protection of an inert gas; the third demethylation reaction is preferably carried out under the protection of an inert gas; wherein the inert gas is preferably nitrogen.

[0113] Furthermore, the present invention discloses a pharmaceutical composition comprising the compound represented by the above-mentioned formula I or any one of its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs, and one or more pharmaceutically acceptable carriers.

[0114] Specifically, the dosage form of the pharmaceutical composition is one or more of subcutaneous injection, intradermal injection, spray, powder aerosol, external solution, lotion, liniment, ointment, plaster, paste, and patch.

[0115] Furthermore, the present invention discloses a pharmaceutical external ointment with the compound represented by the above formula I or its stereoisomer, geometric isomer, tautomer, nitrogen oxide, hydrate, solvate, pharmaceutically acceptable salt or prodrug as an active ingredient, which comprises an oil phase, an aqueous phase and an active ingredient, and the weight percentage of each material is:

[0116] Oil phase:

[0117] Light liquid paraffin 0.6-3%, preferably 0.8-2%, more preferably 1.00%;

[0118] Hexadecanol 1.5-7.5%, preferably 2.0-5.0%, more preferably 2.54%;

[0119] Stearic acid 2.6-5.0%, preferably 3.2-4.4%, more preferably 3.80%;

[0120] Aqueous phase:

[0121] Glycerol 5.4-27%, preferably 7.2-18%, more preferably 8.92%;

[0122] Triethylamine alcohol 0.27-1.38%, preferably 0.35-0.92%, more preferably 0.46%;

[0123] Butylparaben 0.06-3%, preferably 0.08-2%, more preferably 0.10%;

[0124] Active ingredient 0.6-12%, preferably 0.8-8%, more preferably 1% to 4%;

[0125] The balance is purified water;

[0126] The active ingredient is the compound represented by the above formula I or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs.

[0127] Furthermore, the present invention discloses a method for preparing the above-mentioned external ointment, comprising the following steps:

[0128] (1) dissolving the active ingredient in part of purified water to obtain a drug solution; mixing light liquid paraffin, hexyl alcohol, and stearic acid and heating and melting them to obtain an oil phase; mixing glycerin, triethylamine alcohol, butyl hydroxybenzoate, and the remaining purified water and heating and melting them to obtain a water phase;

[0129] (2) adding the drug solution obtained in step (1) to the aqueous phase and mixing to obtain an aqueous phase containing the drug; adding the aqueous phase containing the drug to the oil phase and stirring until the temperature cools to room temperature.

[0130] Specifically, the heating and melting is performed at a temperature of 65 to 70°C.

[0131] The use of the compound represented by the above formula I or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs, or the above pharmaceutical compositions, or the above pharmaceutical external ointments in the preparation of TRPV3 inhibitors is also within the scope of protection of the present invention.

[0132] The use of the compound represented by the above-mentioned formula I or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs, or the above-mentioned pharmaceutical compositions, or the above-mentioned pharmaceutical external ointments in the preparation of drugs for treating TRPV3-mediated diseases is also within the protection scope of the present invention; preferably, the treatment of TRPV3-mediated diseases is to treat TRPV3-mediated diseases by inhibiting the increase in calcium ion influx caused by TRPV3 activation; further preferably, the TRPV3-mediated diseases are one or more diseases selected from pruritus, alopecia, atopic dermatitis, psoriasis, and ulcerative colitis, and further preferably, the TRPV3-mediated diseases are psoriasis; the treatment of TRPV3-mediated psoriasis is to improve psoriasis-like lesions, psoriatic arthritis or psoriasis pain.

[0133] The use of the compound represented by the above formula I or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs, or the above pharmaceutical compositions, or the above pharmaceutical external ointments in the preparation of drugs for treating itching, atopic dermatitis and pain caused by kidney disease is also within the scope of protection of the present invention.

[0134] The use of the compound represented by the above formula I or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs, or the above pharmaceutical compositions, or the above pharmaceutical external ointments in the preparation of drugs for treating kidney disease is also within the scope of protection of the present invention.

[0135] The use of the compound represented by the above formula I or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs, or the above pharmaceutical compositions, or the above pharmaceutical external ointments in the preparation of anti-fibrosis drugs is also within the scope of protection of the present invention.

[0136] It should be pointed out that the YHQ described in the present invention is baicalein.

[0137] The purity of the intermediates prepared in each step in the embodiments of the present invention is above 95%.

[0138] Unless otherwise specified, the term "alkyl" as used herein includes branched and straight chain saturated aliphatic hydrocarbon groups having a specified number of carbon atoms, including all isomers. Common abbreviations for alkyl groups, such as methyl, can be represented by "Me" or CH3, ethyl, can be represented by "Et" or CH2CH3, propyl, can be represented by "Pr" or CH2CH2CH3, butyl, can be represented by "Bu" or CH2CH2CH2CH3, etc. For example, "C 1-4 "C1-C4 alkyl" refers to a straight or branched chain alkyl group having a specified number of carbon atoms, including all isomers. 1-4 Alkyl includes n-, iso-, sec- and tert-butyl, n- and iso-propyl, ethyl and methyl. 1-10 "Alkyl" and the like have similar meanings.

[0139] The term "alkoxy" refers to straight and branched chain alkyl groups having the indicated number of carbon atoms attached through an oxygen bridge.

[0140] The term "halogen" (or "halo") refers to fluorine, chlorine, bromine, and iodine (alternatively referred to as fluoro (F), chloro (Cl), bromo (Br), and iodo (I)).

[0141] The term "aryl" refers to aromatic mono- and polycyclic carbon ring systems wherein the individual carbon rings in the polycyclic system are fused or linked to each other by single bonds. Typical aryl groups include phenyl, naphthyl and biphenylene.

[0142] The term "heterocycle" refers to a ring structure composed of carbon atoms and non-carbon atoms, such as nitrogen, oxygen and sulfur. Typical heterocyclic groups include pyridine, quinoline, tropane, phenothiazine, benzodiazepine, furan, pyrazolone and pyrimidine.

[0143] The term "aromatic heterocycle" refers to a 5- or 6-membered monocyclic aromatic ring or a 7- to 12-membered bicyclic ring consisting of carbon atoms and one or more heteroatoms selected from N, O and S. Examples of aromatic heterocycles include pyridyl, pyrrolyl, pyrazinyl, pyrimidinyl, pyridazinyl, thienyl (or thiophenyl), thiazolyl, furanyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl and thiadiazolyl, benzotriazolyl, indolyl, isoindolyl, indazolyl, indolinyl, isoindolinyl, quinoxalinyl, quinazolinyl, cinnolinyl, chromanyl, isochromanyl, tetrahydroquinolyl, quinolyl, tetrahydroisoquinolyl, isoquinolyl, 2,3-dihydrobenzofuranyl, 2,3-dihydrobenzo-1,4-dienyl, imidazo(2,1-b)(1,3)thiazole and benzo-1,3-dioxolyl.

[0144] The aryl group in the term "substituted aryl" is as defined above. When the substituent of the substituted aryl group is not specified, the substituent may be selected from the following groups, including but not limited to: halogen, C1-C 20 Alkyl, CF3, NH2, N(C1-C6 alkyl)2, NO2, oxo, CN, N3, -OH, -O(C1-C6 alkyl), C3-C 10 Cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, (C0-C6 alkyl)S(O) 0-2 -, Aryl-S(O) 0-2 -, (C0-C6 alkyl) S (O) 0-2 (C0-C6 alkyl)-, (C0-C6 alkyl)C(O)NH-, H2N-C(NH)-, -O(C1-C6 alkyl)CF3, (C0-C6 alkyl)C(O)-, (C0-C6 alkyl)OC(O)-, (C0-C6 alkyl)2NC(O)-(C0-C6 alkyl)O(C1-C6 alkyl)-, (C0-C6 alkyl)C(O) 1-2 (C0-C6 alkyl)-, (C0-C6 alkyl)OC(O)NH-, aryl, aralkyl, heteroaryl, heterocyclylalkyl, halogen-aryl, halogen-aralkyl, halogen-heterocycle, halogen-heterocyclylalkyl, cyano-aryl, cyano-aralkyl, cyano-heterocycle and cyano-heterocyclylalkyl. The term "substituted phenyl" has a similar definition.

[0145] The term "pharmaceutically acceptable salt" refers to a salt prepared from a pharmaceutically acceptable non-toxic base or acid. When the compound of the present invention is acidic, its corresponding salt can be easily prepared from an inorganic base or an organic base. Salts derived from such inorganic bases include salts of aluminum, ammonium, calcium, copper (copper and cuprous), iron, ferrous, lithium, magnesium, manganese (manganese and manganous), potassium, sodium, zinc, etc. Preferred are salts of ammonium, calcium, magnesium, potassium and sodium. Salts prepared from organic bases include primary, secondary and tertiary amines derived from natural and synthetic sources. Pharmaceutically acceptable organic non-toxic bases that can form salts include arginine, betaine, caffeine, choline, N,N'-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucosamine, glucosamine, histidine, hydrabamine, isopropylamine, dicyclohexylamine, lysine, methylglucamine, morpholine, piperazine, piperidine, polyamine resins, procaine, purines, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine, etc. When the compound of the present invention is basic, its corresponding salt can be easily prepared from inorganic or organic acids. Such acids include, for example, acetic acid, benzenesulfonic acid, benzoic acid, camphorsulfonic acid, citric acid, ethanesulfonic acid, fumaric acid, gluconic acid, glutamic acid, hydrobromic acid, hydrochloric acid, isethionic acid, lactic acid, maleic acid, malic acid, mandelic acid, methanesulfonic acid, mucic acid, nitric acid, pamoic acid, pantothenic acid, phosphoric acid, succinic acid, sulfuric acid, tartaric acid, p-toluenesulfonic acid, and the like.

[0146] The term "solvate" refers to a complex of variable stoichiometry formed by a solute (i.e., a compound of Formula I) or a pharmaceutically acceptable salt thereof and a solvent that does not interfere with the biological activity of the solute. Examples of solvents include, but are not limited to, water, ethanol, and acetic acid. When the solvent is water, the solvate is referred to as a hydrate. Hydrates include, but are not limited to, hemihydrates, monohydrates, one and a half hydrates, dihydrates, and trihydrates.

[0147] The present invention provides a pharmaceutical composition comprising a compound represented by Formula I or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs, and one or more pharmaceutically acceptable carriers, diluents, and excipients.

[0148] In pharmaceutical compositions, the term "composition" includes products comprising active ingredients and inert ingredients constituting carriers (pharmaceutically acceptable excipients), as well as any product obtained directly or indirectly by the combination, complexation or aggregation of two or more ingredients, or the decomposition of one or more ingredients, or other types of reactions or interactions of one or more ingredients. Therefore, the pharmaceutical compositions of the present invention include any composition prepared by mixing a compound of formula I, other active ingredients and a pharmaceutically acceptable excipient.

[0149] The pharmaceutical composition of the present invention comprises the compound shown in the formula I as active ingredient (or its pharmaceutically acceptable salt or solvate), pharmaceutically acceptable carrier and optional other therapeutic ingredients or adjuvant.The pharmaceutical composition includes compositions suitable for oral, rectal, topical and parenteral (including subcutaneous, intramuscular and intravenous) administration, although the most suitable approach in any particular case depends on the nature and severity of the disease to which the active ingredient is given.The pharmaceutical composition can be prepared by any method known to the field of pharmacy.

[0150] The active ingredient can be administered orally in a solid dosage form or a liquid dosage form, such as capsules, tablets, lozenges, lozenges, granules and powders, and the liquid dosage form is such as elixirs, syrups, emulsions, dispersions and suspensions. The active ingredient can also be administered parenterally in a sterile liquid dosage form such as a dispersion, suspension or solution. Other dosage forms that can be used to administer the active ingredient include subcutaneous injections, intradermal injections, external solutions, lotions, liniments, pastes, ointments, creams, drops, transdermal patches or powders for topical administration; ophthalmic solutions or suspension forms for eye administration, i.e. eye drops; sprays or powder compositions for inhalation or intranasal administration, or creams, ointments, sprays or suppositories for rectal or vaginal administration. Gelatin capsules contain active ingredients and powdered carriers, such as lactose, starch, cellulose derivatives, magnesium stearate, stearic acid, etc. Similar diluents can be used to prepare compressed tablets. Tablets and capsules can be prepared into sustained-release products to provide sustained release of medicine within a few hours. Compressed tablets can be coated with sugar or film to cover any unpleasant taste and protect the tablet from air, or can be enteric-coated for selective disintegration in the gastrointestinal tract. Liquid dosage forms for oral administration can include coloring agents and flavoring agents to increase patient acceptance. Generally speaking, water, suitable oil, saline, dextrose (glucose) aqueous solution and related sugar solution and glycols such as propylene glycol or polyethylene glycol are suitable carriers of parenteral solutions. The solution for parenteral administration preferably includes water-soluble salts of active ingredients, suitable stabilizers and buffer substances used as required. Antioxidants such as sodium bisulfite, sodium sulfite or ascorbic acid alone or in combination are suitable stabilizers. Citric acid and its salts and sodium EDTA can also be used. In addition, parenteral solutions can also include preservatives, such as benzalkonium chloride, methylparaben or propylparaben and chlorobutanol. For inhalation administration, the compound of the present invention can be conveniently delivered in the form of a spray from a pressurized package or a nebulizer. The compound can also be delivered in the form of a powder for preparation, and the powder composition can be inhaled with the help of a powder inhaler device. The preferred delivery system for inhalation is a metered dose inhalation (MDI) aerosol, which can be formulated into a suspension or solution of the compound of formula I, II in a suitable propellant, such as a fluorocarbon or a hydrocarbon. For eye administration, ophthalmic preparations can be prepared with a solution or a suspension of a suitable weight percentage of the compound of formula I in a suitable ophthalmic carrier, thereby keeping the compound in contact with the surface of the eye for enough time to allow the compound to penetrate into the cornea and the inner area of ​​the eye.

[0151] Useful pharmaceutical dosage forms for administering the compounds of the present invention include, but are not limited to, hard and soft gelatin capsules, tablets, parenteral injection solutions, and oral suspensions.

[0152] When the compounds of the present invention are administered stepwise or in combination with other therapeutic agents, the same dosage forms as described above may be used. When the drugs are administered in physical combination, the dosage form and route of administration should be selected based on the compatibility of the combined drugs. The compounds of the present invention may be administered as the sole active ingredient or in combination with a second active ingredient, including active ingredients known to be useful for increasing erythropoietin levels in patients.

[0153] Beneficial effects:

[0154] (1) The present invention provides a new class of compounds that can be used as TRPV3 inhibitors, which can inhibit the abnormal increase in calcium ion influx caused by TRPV3 activation, and can effectively treat and prevent TRPV3-related diseases, providing new ideas for diseases such as itching, hair loss, atopic dermatitis, psoriasis, and ulcerative colitis.

[0155] (2) The compounds provided by the present invention have better solubility under the premise of having a comparable inhibition rate on calcium ion influx; at the same time, the compounds provided by the present invention have an inhibitory effect on the proliferation of human hepatic stellate cells LX-2 and have an anti-fibrotic effect. DETAILED DESCRIPTION

[0156] The following non-limiting examples can enable those skilled in the art to more fully understand the present invention, but do not limit the present invention in any way. The following content is merely an exemplary description of the scope of protection claimed in this application, and those skilled in the art can make various changes and modifications to the invention of this application based on the disclosed content, and they should also fall within the scope of protection claimed in this application.

[0157] The present invention is further described below by way of specific examples. Unless otherwise specified, the various chemical reagents used in the examples of the present invention are obtained through conventional commercial channels.

[0158] Example 1: Preparation of 5,7,4'-trihydroxy-8-isopropylflavone

[0159]

[0160] Step 1: Preparation of 2-isopropyl-3,5-dimethoxyphenol

[0161] Substitution reaction: 3,5-dimethoxyphenol (8g, 51.89mmol), chloroform (60mL), 2-chloropropane (4.9g, 62.26mmol) and aluminum chloride (18.7g, 140.24mmol) were added to the reaction bottle in sequence, stirred in an ice-water bath at 0°C for 0.5h, slowly warmed to room temperature, and the substitution reaction was carried out for 4h until the reaction of the raw materials was complete as monitored by TLC. After the reaction, the reaction solution was cooled to room temperature, 20mL of purified water was slowly added dropwise to quench the reaction, extracted with dichloromethane, washed with saturated brine 2 to 3 times, separated the organic phase, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the eluent was ethyl acetate: petroleum ether (volume ratio of 1:40), and 4.0g of light yellow solid was obtained by column chromatography, with a yield of 39%. The mass spectrum data of the product 2-isopropyl-3,5-dimethoxyphenol: MS m / z (ESI) [M+H] + :197.11.

[0162] Step 2: Preparation of 2-hydroxy-3-isopropyl-4,6-dimethoxyacetophenone

[0163] Substitution reaction: 2-isopropyl-3,5-dimethoxyphenol (2.4 g, 12.22 mmol), acetyl chloride (1.2 g, 14.66 mmol), chloroform (20 mL, obtained by redistillation), and aluminum chloride (3.2 g, 24.46 mmol) prepared in step 1 were added to the reaction bottle in sequence, stirred at room temperature, and the substitution reaction was carried out for 4 h until the reaction of the raw materials was complete as monitored by TLC. After the reaction was completed, 20 mL of purified water was slowly added dropwise in an ice-water bath to quench the reaction, the layers were allowed to stand, the organic phase was separated, the saturated brine was washed 2 to 3 times, the organic phase was dried over anhydrous sodium sulfate, and the eluent was ethyl acetate: petroleum ether (volume ratio of 1:30), and 1 g of a light yellow oil was obtained by column chromatography with a yield of 34%. The mass spectrum data of the product 2-hydroxy-3-isopropyl-4,6-dimethoxyacetophenone: MS m / z (ESI) [M+H] + :239.28.

[0164] Step 3: Preparation of 1-(2-hydroxy-3-isopropyl-4,6-dimethoxyphenyl)-3-(4-methoxyphenyl)propane-1,3-dione

[0165] There are two reactions in this step: the compound first undergoes an esterification reaction to generate 2-acetyl-6-isopropyl-3,5-dimethoxyphenyl-4-methoxybenzoate, and then the compound undergoes a rearrangement.

[0166] Esterification reaction and rearrangement: 2-hydroxy-3-isopropyl-4,6-dimethoxyacetophenone (0.8 g, 3.36 mmol), tetrahydrofuran (15 mL), sodium hydroxide (0.3 g, 7.5 mmol), and 4-methoxybenzoyl chloride (0.6 g, 3.50 mmol) prepared in step 2 were added to the reaction flask in sequence, and nitrogen was replaced three times, and stirred at room temperature until the raw material reaction was complete as monitored by TLC. After the reaction, dilute acetic acid was added dropwise to adjust the pH to neutral, extracted with dichloromethane, washed with saturated brine 2 to 3 times, separated the organic phase, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the eluent was ethyl acetate: petroleum ether (volume ratio of 1:8), and column chromatography was performed to obtain 1.2 g of a yellow solid with a yield of 96%. Mass spectral data of the product 1-(2-hydroxy-3-isopropyl-4,6-dimethoxyphenyl)-3-(4-methoxyphenyl)propane-1,3-dione: MS m / z(ESI)[M+H] + :373.16.

[0167] Step 4: Preparation of 5,7,4'-trimethoxy-8-isopropylflavone

[0168] Ring-closing reaction: 1-(2-hydroxy-3-isopropyl-4,6-dimethoxyphenyl)-3-(4-methoxyphenyl)propane-1,3-dione (1.0 g, 2.68 mmol), acetic acid (10 mL), and concentrated sulfuric acid (0.1 g, 1 mmol) prepared in step 3 were added to the reaction bottle in sequence, and the ring-closing reaction was carried out by stirring at room temperature for 3 h. The peak ratio of the raw material monitored by LC-MS was less than or equal to 0.1%. Then 15 mL of purified water was added, and then 20 mL of dichloromethane solution was added and stirred, and the mixture was allowed to stand for stratification. The organic phase was washed with saturated brine 2 to 3 times, the organic phase was separated, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The eluent was ethyl acetate: petroleum ether (volume ratio of 1:10) / methanol: dichloromethane (volume ratio of 1:40), and 0.8 g of yellow solid was obtained by column chromatography with a yield of 84%. Mass spectral data of the product 5,7,4'-trimethoxy-8-isopropylflavone: MS m / z(ESI)[M+H] + :355.15.

[0169] Step 5: Preparation of 5,7,4'-trihydroxy-8-isopropylflavone

[0170] Demethylation reaction: 5,7,4'-trimethoxy-8-isopropylflavone (0.5 g, 1.41 mmol) prepared in step 4 and 2 mol / L dichloromethane solution of boron tribromide (10 mL, containing 20 mmol of BBr3) were added to the reaction bottle in sequence, and the mixture was protected by nitrogen. The demethylation reaction was carried out by stirring at room temperature for 4 h. The peak ratio of the raw material was less than or equal to 0.1% as monitored by LC-MS. The solvent was removed by rotation, and 4 mL of methanol was slowly added to quench the mixture. The methanol was rotated to dryness, and 1.5 mL of methanol was added to dissolve the residue. Then 3 mL of purified water was slowly added and stirred for 2 h. A large amount of orange-red solid was precipitated. The filter cake was vacuum dried at 50 ° C to obtain 0.36 g of orange-yellow solid with a yield of 82%. The product 5,7,4'-trihydroxy-8-isopropylflavone was recorded as compound 1, mp: 186.0-186.5 ° C, and the nuclear magnetic resonance data and mass spectrometry data are as follows:

[0171] 1 H-NMR (400MHz, DMSO) δ: 13.04 (s, 1H, OH), 10.68 (s, 1H, OH), 10.37 (s, 1H, OH), 7.90 (d, J=8.0Hz, 2H, ArH), 6.96 (d, J=8.0Hz, 2H, ArH), 6.75 (s, 1H, ArH), 6.28 (s, 1H, ArH), 3.66 (m, 1H, CH), 1.36 (d, J=8.0Hz, 6H, CH); 13 C-NMR (101MHz, DMSO) 182.30, 163.94, 162.04, 161.22, 159.11, 154.77, 128.43, 121.71, 116.22, 111.80, 103.91, 102.86, 98.96, 23.56, 20.82; MS m / z(ESI)[M+H] + :313.10.

[0172] Example 2: Preparation of 2-(3-fluoro-4-hydroxyphenyl)-6,7-dihydroxyquinolin-4(1H)-one

[0173]

[0174] Step 1: Preparation of N-(2-acetyl-4,5-dimethoxyphenyl)-3-fluoro-4-methoxybenzamide

[0175] Acylation reaction: 2-amino-4,5-dimethoxyacetophenone (500 mg, 2.56 mmol, Mw = 195.218), tetrahydrofuran (20 mL), 3-fluoro-4-methoxybenzoyl chloride (724.2 mg, 3.84 mmol, Mw = 188.582), and triethylamine (1 mL, 0.728 g, 7.19 mmol, Mw = 101.19) were added to the reaction bottle in sequence, and nitrogen was replaced three times. The temperature was raised to 65 ° C and the acylation reaction was carried out for 4 hours until the raw material reaction was complete as monitored by TLC. After the reaction, the reaction solution was cooled to room temperature, and 20 mL of purified water was slowly added to quench the reaction, extracted with dichloromethane, washed with saturated brine 2 to 3 times, separated the organic phase, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The eluent was dichloromethane: ethyl acetate: petroleum ether (volume ratio of 1:1:3), and column chromatography was performed to obtain 778 mg of a light yellow solid with a yield of 87.5%. The mass spectrum data of the product N-(2-acetyl-4,5-dimethoxyphenyl)-3-fluoro-4-methoxybenzamide (Mw=347.342): MS m / z(ESI)[M+H] + :348.34.

[0176] Step 2: Preparation of 6,7-dimethoxy-2-(3-fluoro-4-methoxyphenyl)quinolin-4(1H)-one

[0177] Cyclization reaction: N-(2-acetyl-4,5-dimethoxyphenyl)-3-fluoro-4-methoxybenzamide (330 mg, 0.95 mmol, Mw=347.342) prepared in step 1, 1,4-dioxane (15 mL), sodium hydroxide (250 mg, 6.25 mmol, Mw=40.00) were added to the reaction bottle in sequence, and the nitrogen was replaced three times, and the temperature was raised to 110°C for intramolecular cyclization reaction for 4 hours until the reaction of the raw materials was complete as monitored by TLC. After the reaction, the reaction solution was cooled to room temperature, diluted acetic acid was added dropwise to adjust the pH to neutral, extracted with dichloromethane, washed with saturated brine 2 to 3 times, the organic phase was separated, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The eluent was dichloromethane: methanol (volume ratio of 100:7 to 100:10), and 252 mg of white solid was obtained by column chromatography with a yield of 80.7%. Mass spectral data of the product 6,7-dimethoxy-2-(3-fluoro-4-methoxyphenyl)quinolin-4(1H)-one (Mw=329.327): MS m / z (ESI) [M+H] + :330.31.

[0178] Step 3: Preparation of 6,7-dihydroxy-2-(3-fluoro-4-hydroxyphenyl)quinolin-4(1H)-one

[0179] Demethylation reaction: 6,7-dimethoxy-2-(3-fluoro-4-methoxyphenyl)quinoline-4(1H)-one (210 mg, 0.64 mmol, Mw=329.327) prepared in step 2 and 2 mol / L dichloromethane solution of boron tribromide (2 mL, containing BBr34 mmol) were added to the reaction bottle in sequence, nitrogen protection, stirring at room temperature for demethylation reaction for 4 h, LC-MS monitoring showed that the raw material peak ratio was less than or equal to 0.1%, the solvent was removed by rotation, 4 mL of methanol was slowly added to quench, methanol was spin-dried, 1.5 mL of methanol was added to dissolve the residue, and then 3 mL of purified water was slowly added and stirred for 2 h. A large amount of orange-red solid precipitated, filtered, and the filter cake was dried under vacuum at 50 ° C to obtain 108.9 mg of the product. The product 2-(3-fluoro-4-hydroxyphenyl)-6,7-dihydroxyquinoline-4(1H)-one (C 15 H 10 FNO4, Mw = 287.246) is recorded as compound 2, mp: 232.0-233.5 ° C, NMR data and mass spectrum data are as follows:

[0180] 1 H NMR (400MHz, d-DMSO) δ: 13.75 (s, 1H, NH), 11.28 (s, 1H, OH), 10.91 (s, 1H, OH), 10.56 (s, 1H, OH), 7.84 (d, J=8.0Hz, 1H , ArH), 7.60 (d, J=4.0Hz, 1H, ArH), 7.51 (s, 1H, ArH), 7.46 (s, 1H, ArH), 7.22 (t, J=6.0Hz, 1H, ArH), 7.06 (s, 1H, ArH); 13 C-NMR (101MHz, DMSO)161.32, 149.47, 146.70, 144.98, 143.42, 142.89, 131.42, 120.30, 117.87, 113.41, 111.23, 108.50, 99.48, 97.86, 96.46; m / z(ESI)[M+H] + :288.06.

[0181] Example 3: Preparation of 6,7-dihydroxy-2-(4-isopropylphenyl)quinolin-4(1H)-one

[0182]

[0183] The compound of this example was prepared according to the preparation method of Example 2, and the material ratios between the materials remained unchanged, except that 3.84 mmol of 4-isopropylbenzoyl chloride was used to replace 3-fluoro-4-methoxybenzoyl chloride to obtain 128.9 mg of the product. The product 6,7-dihydroxy-2-(4-isopropylphenyl)quinoline-4(1H)-one (C 18 H 17 NO3, Mw = 295.338) is recorded as compound 3, mp: 125.0-126.5 ° C, NMR data and mass spectrum data are as follows:

[0184] 1 H-NMR (400MHz, DMSO) δ: 13.98 (s, 1H, NH), 11.28 (s, 1H, OH), 10.57 (s, 1H, OH), 7.84 (d, J=8.0Hz, 2H, ArH), 7.60 (s, 1H, Ar H), 7.54 (d, J=8.0Hz, 2H, ArH), 7.49 (s, 1H, ArH), 7.16 (s, 1H, ArH), 3.01 (p, J=6.8Hz, 1H, CH), 1.25 (d, J=6.8Hz, 6H, CH); 13 CS m / z(ESI)[M+H] + :296.12.

[0185] Example 4: 2-(3,4-dichlorophenyl)-6,7-dihydroxyquinolin-4(1H)-one

[0186]

[0187] The preparation was carried out according to the preparation method of Example 2, and the material ratio between the materials remained unchanged, except that 3.84 mmol of 3,4-dichlorobenzoyl chloride was used to replace 3-fluoro-4-methoxybenzoyl chloride, and 120.3 mg of the product was obtained. The product 2-(3,4-dichlorophenyl)-6,7-dihydroxyquinoline-4(1H)-one (C 15 H9C l2 NO3, Mw = 322.141) is recorded as compound 4, mp: 206.2-207.5 ° C, NMR data and mass spectrum data are as follows:

[0188] 1H-NMR (400MHz, DMSO) δ: 11.01 (s, 1H, NH), 10.32 (s, 1H, OH), 8.21 (d, 1H, OH), 7.88 (m, 3H, ArH), 7.46 (s, 1H, ArH), 7.39 (s, 1H, ArH), 6.87 (s, 1H, ArH); 13 C-NMR (101MHz, DMSO)168.99, 153.90, 147.74, 147.36, 136.69, 133.96, 13 3.57, 132.02, 131.44, 129.98, 128.37, 115.46, 105.38, 103.57, 103.12; MS m / z(ESI)[M+H] + :322.99.

[0189] Example 5: Preparation of N-(2-acetyl-4,5-dihydroxyphenyl)-3-fluoro-4-hydroxybenzamide

[0190]

[0191] The preparation was carried out according to the preparation methods of step 1 (acylation reaction) and step 3 (demethylation reaction) in Example 2, with the material ratio between the materials remaining unchanged, 3.84 mmol of 3-fluoro-4-methoxybenzoyl chloride was used to carry out acylation reaction with 2-amino-4,5-dimethoxyacetophenone, followed by demethylation reaction, to obtain 201.6 mg of the product. The product N-(2-acetyl-4,5-dihydroxyphenyl)-3-fluoro-4-hydroxybenzamide (C 15 H 12 FNO5, Mw = 305.261) is recorded as compound 5, mp: 292.0-293.5 ° C, NMR data and mass spectrum data are as follows:

[0192] 1 H NMR (400MHz, d-DMSO) δ: 12.62 (s, 1H, OH), 10.76 (s, 1H, OH), 10.48 (s, 1H, OH), 9.16 (s, 1H, N H), 8.29 (s, 1H, ArH), 7.64 (m, 2H, ArH), 7.42 (s, 1H, ArH), 7.12 (t, 1H, ArH), 2.56 (s, 3H, CH); 13CS m / z(ESI)[M+H] + :306.07.

[0193] Example 6: N-(2-acetyl-4,5-dihydroxyphenyl)-3,4-dichlorobenzamide

[0194]

[0195] The preparation was carried out according to the preparation methods of step 1 (acylation reaction) and step 3 (demethylation reaction) in Example 2, and the material ratio between the materials remained unchanged, except that 3.84 mmol of 3,4-dichlorobenzoyl chloride was used to replace 3-fluoro-4-methoxybenzoyl chloride, and acylation reaction was carried out with 2-amino-4,5-dimethoxyacetophenone, followed by demethylation reaction, to obtain 191.6 mg of product. The product N-(2-acetyl-4,5-dihydroxyphenyl)-3,4-dichlorobenzamide (C 15 H 11 C l2 NO4, Mw = 340.156), recorded as compound 6, the NMR data and mass spectrum data are as follows:

[0196] 1 H-NMR (400MHz, DMSO) δ: 12.72 (s, 1H, OH), 10.51 (s, 1H, OH), 9.19 (s, 1H, NH), 8.25 ( s, 1H, ArH), 8.06 (s, 1H, ArH), 7.85 (s, 2H, ArH), 7.43 (s, 1H, ArH), 2.56 (s, 3H, CH); 13 CS m / z(ESI)[M+H]+:341.00.

[0197] Example 7: Water solubility test

[0198] 1. Reagents and Consumables

[0199] Table 1 Reagents and consumables

[0200]

[0201]

[0202] 2. Experimental Methods

[0203] Use DMSO to prepare 10mmol / L stock solutions of test compound 1-6 (prepared in Examples 1 to 6, respectively), YHQ (scutellaria baicalensis, homemade) and control standard progesterone. Take 30μL of 10mmol / L test stock solution, add it to a 96-well plate in turn, and then add 970μL of phosphate buffer with a pH value of 7.4. Stir for 2 hours at 25°C and 1100r / min. Filter, transfer 5μL of filtrate to a new sample plate, add 5μL DMSO and 490μL 50% ACN (acetonitrile) solution. The standard solution is prepared with a phosphate buffer with a pH value of 7.4 to make the final concentration of progesterone 3μmol / L. The saturated solubility of the test compound is calculated by qualitative and quantitative analysis of the standard peak of known concentration by liquid mass spectrometry as shown in Table 2.

[0204] Table 2 Solubility of each compound

[0205]

[0206] Example 8: Biological Test

[0207] 1. Calcium ion concentration detection

[0208] 1.1 Reagents and consumables

[0209] Table 3 Reagents and consumables

[0210]

[0211]

[0212] 1.2 Test methods

[0213] (1) Preparation of compound stock solutions

[0214] Preparation of stock solutions of compounds 1-6: Accurately weigh each compound powder (prepared in Examples 1 to 6, respectively), prepare 1 mM stock solutions with DMSO, and store at -80°C after aliquoting. The shelf life is 1 year.

[0215] Preparation of TRPV3 agonist 2-APB stock solution: weigh 1.3505 mg of 2-APB powder, add 200 μL of DMSO solution to prepare a 30 mM stock solution, and store it at -80°C after aliquoting.

[0216] Preparation of TRPV3 inhibitor YHQ (baicalein, homemade) stock solution: weigh 1.7174 mg baicalein powder, add 200 μL DMSO solution to prepare 1 mM stock solution, and store it at -80°C after aliquoting.

[0217] (2) Buffer preparation

[0218] Preparation of extracellular solution for TRPV3 calcium signal detection (abbreviated as: extracellular solution buffer): After mixing the materials in Table 4, adjust the pH to 7.4 with NaOH to obtain.

[0219] Table 4 Amount of each component in 1000mL of external solution for TRPV3 calcium signal detection

[0220]

[0221] Preparation of calcium fluorescent dye: Add 228 μL DMSO to 1 mg Fluo-4 powder to prepare a 4 mM stock solution. Wrap in tin foil to protect from light, vortex, and dispense into 200 μL centrifuge tubes, 10 μL per tube.

[0222] Preparation of PBS buffer: Mix the components in Table 5, and then adjust the pH to 7.2-7.4 with HCl and NaOH.

[0223] Table 5 Components and dosage in 1000mL PBS buffer

[0224]

[0225] (3) Cell culture

[0226] HEK-293 cells were cultured in DMEM complete medium containing 10% FBS + 1% P / S + 1% HEPES at 37°C in a 5% CO2 incubator. When the cell density reached 80%, PBS buffer (pH = 7.2-7.4), trypsin-EDTA digestion solution, and HEK-293 culture medium were preheated and filtered in a biosafety cabinet, the culture medium in the bottle was discarded, the remaining culture medium was washed twice with PBS and discarded, trypsin-EDTA digestion solution was added to digest the cells until the cells became round, and the trypsin-EDTA digestion solution was discarded after the cytoplasm shrank, DMEM complete medium was added to terminate the digestion and the cells at the bottom of the bottle were blown off, the cell suspension was transferred to a small dish (density 50%-60%), DMEM complete medium was added and slowly blown evenly, and placed in an incubator for 6 hours before cell transfection.

[0227] (4) Cell transfection

[0228] Preheat PBS buffer (pH = 7.2-7.4) and reduced serum culture medium (opti-MEM) and filter them in a biosafety cabinet. Discard the original culture medium in the small dish (finally obtained in "(3) Cell culture"), wash twice with PBS, add opti-MEM and culture in a cell culture incubator for 1 hour. TM The transfection system was prepared according to the instruction manual of 2000 transfection reagent. After 1 hour, the solution mixed with plasmid and transfection reagent was added dropwise into the small dish. After culturing in a cell culture incubator for 6-8 hours, it was replaced with normal DMEM complete medium for subsequent plating experiments.

[0229] (5) Cell plating

[0230] 96-well plate polylysine pre-coating: Use sterile water to prepare the polylysine mother solution to 10 mg / mL, divide it into 0.2 mL centrifuge tubes, 10 μL per tube, and store it in an ultra-low temperature refrigerator. When using, take the polylysine mother solution out of the ultra-low temperature refrigerator, add sterile water to dilute it to a final concentration of 10 μg / mL, filter the diluted polylysine solution, and coat 60 μL / well of the liquid in a 96-well black-walled bottom transparent plate, and incubate it in a 37°C, 5% CO2 incubator for 1 hour. Before inoculating cells, aspirate and discard the polylysine solution in the well.

[0231] 14 h before the real-time fluorescence experiment, the cells were washed with PBS, and TRPV3-HEK 293 transiently transfected cells were digested with trypsin-EDTA solution and inoculated into a 96-well black-bottom transparent plate coated with poly-L-lysine. 100 μL / well (i.e., 2 × 10 4 cells / well.

[0232] (6) Add 60 μL of dye (“calcium fluorescent dye” prepared under “(2) buffer preparation”), place the cell plate back into the incubator, and load the dye for 60 min.

[0233] (7) Remove the cell plate and add 140 μL of extracellular solution buffer (prepared under “(2) Buffer preparation”) to each well to make the volume in the well 200 μL. Wash with 150 μL of extracellular solution buffer and repeat 3-4 times. After aspirating 150 μL of solution for the last time, add 100 μL of extracellular solution buffer to make the volume of extracellular solution in each well 150 μL. After washing, place the cell plate in the Read position of the FLIPR and stabilize for 5 minutes.

[0234] The test compound (the mixture of the inhibitor YHQ, compound 1-6 and the extracellular fluid, respectively, so that the final concentration is 1.0 μmol / L) and the agonist (the mixture of 2-APB and the extracellular fluid, so that the final concentration is 30 μmol / L) were added at 60 seconds and 360 seconds after the start of signal collection, respectively, and the signal collection interval was 1 second. The fluorescence values ​​at different time points were recorded at a wavelength of 535 nm. The fluorescence values ​​were read from the Screen work 4.0 software (Molecular device, CA, USA), and the data were exported and processed to obtain the inhibition rate of the test compound. The results are shown in Table 6 below:

[0235] Table 6 Inhibition rate of each compound on calcium ion

[0236] Compound No. Inhibition rate % (concentration 1.0 μM) Inhibitor YHQ (scutellaria baicalensis) (70.24±0.91)％ Compound 1 (prepared in Example 1) (70.78±0.40)％ Compound 2 (prepared in Example 2) (70.49±0.53)％ Compound 3 (prepared in Example 3) (71.82±0.73)％ Compound 4 (prepared in Example 4) (52.63±1.28)％ Compound 5 (prepared in Example 5) (48.35±0.24)％ Compound 6 (prepared in Example 6) (41.87+1.96)％

[0237] As can be seen from the above table, compounds 1 to 3 prepared in the present invention can inhibit calcium ion influx at a single concentration of 1.0 μM and have significant TRPV3 inhibitory activity.

[0238] Example 9: Preparation of a pharmaceutical ointment for external use

[0239] The ointment for external use of the medicine includes an oil phase, an aqueous phase and an active ingredient, and the weight percentage of each material is:

[0240] Oil phase: light liquid paraffin 1.00%; hexadecanol 2.54%; stearic acid 3.80%;

[0241] Aqueous phase: glycerol 8.92%; triethylamine 0.46%; butyl hydroxybenzoate 0.10%;

[0242] The balance is purified water;

[0243] Active ingredients: Compounds 1 to 6 (prepared in Examples 1 to 6, respectively).

[0244] Preparation of medicinal ointment for external use:

[0245] (1) Compounds 1 to 6 were dissolved in part of purified water to obtain a drug solution; 1.00% light liquid paraffin, 2.54% hexadecanol, and 3.80% stearic acid were mixed and heated to melt at 65 to 70° C. to obtain an oil phase; 8.92% glycerol, 0.46% triethylamine alcohol, 0.10% butyl hydroxybenzoate, and the remaining purified water were mixed and heated to melt at 65 to 70° C. to obtain an aqueous phase.

[0246] (2) adding the drug solution obtained in step (1) to the aqueous phase and mixing well to obtain an aqueous phase containing the drug; adding the aqueous phase containing the drug to the oil phase and stirring rapidly until the temperature is cooled to room temperature, thereby preparing a light yellow external ointment containing 1% of compound 1 to 6.

[0247] Example 10: Pharmacological Evaluation of Compounds

[0248] 1. Purpose of the experiment

[0249] The purpose of this experiment is to evaluate the effect of Compounds 1 to 6 (prepared in Examples 1 to 6, respectively) on the efficacy of imiquimod-induced psoriasis in C57BL / 6 mice.

[0250] 2. Experimental Animals

[0251] 25 SPF-grade C57BL / 6 mice were purchased from the Comparative Medicine Center of Yangzhou University [Laboratory Animal Production License No.: SCXK (Su) 2022-0009], 11 weeks old, male mice, weighing 25-30 g. They were kept at room temperature (23 ± 2) °C, humidity 55%, light and dark for 12 h each, with free access to water and food, and adaptive feeding for 7 days. The experiment was approved by the Institute of Health and Environmental Technology of Soochow University (Laboratory Animal Use License No.: SYXK (Su) 2022-0057).

[0252] 3. Information on the test compound and positive control drug

[0253] Imiquimod ointment (Imiquimod, IMQ; batch number: 40220601, Sichuan Mingxin Pharmaceutical Co., Ltd., 50 mg / g); mometasone furoate ointment (Mometasone, Momet; batch number: 221008, Shanghai Pharmaceutical Minhang Co., Ltd., white semisolid ointment, 3 g, 1 mg / g); 1% compound 2 ointment (prepared in Example 9); baicalein (YHQ, homemade).

[0254] 4. Animal grouping and animal model establishment

[0255] Twenty-five SPF-grade C57BL / 6 male mice were shaved of their dorsal hair (an area of ​​about 2×3 cm), and the remaining hair was removed with a depilatory cream. After 24 hours, the mice were randomly divided into 5 groups: blank control group (Control), model group (IMQ), modeling + mometasone furoate group (IMQ+Momet), modeling + YHQ group (IMQ+YHQ), and modeling + 1% compound 2 ointment group (IMQ+Compound 2), with 5 mice in each group.

[0256] Except for the blank control group, the other groups applied 62.5 mg IMQ on the back skin of mice every morning to make models; 8 hours after administration, the same dose of corresponding drug ointment (the dose of each cream was 100 mg, the dose of YHQ was 100 mg) was given for 5 consecutive days.

[0257] Preparation of blank matrix: The component ratio and preparation method of the blank matrix are the same as those of Example 9, except that no active ingredient compound is added.

[0258] 5. Project testing

[0259] (1) On days 1, 3, and 5, and at the same time on the first day of the experiment, the severity of the skin lesions of the mice was scored using the PASI scoring scale from three aspects: erythema, scaling, and skin thickening. The PASI scoring scale ranges from 0 to 4 points, with severity levels being none, mild, moderate, and severe. The total score is the sum of the three indicators (0 to 12 points).

[0260] (2) The skin tissue of the dorsal administration area of ​​the mice was photographed on the 1st, 3rd and 5th day. After the modeling and administration, the skin tissue of the dorsal administration area of ​​the mice was collected, washed with saline, fixed in 4% (mass fraction) paraformaldehyde solution for 48 hours, and then paraffin-embedded. Then, it was cut into 3 μm paraffin sections and stained with conventional hematoxylin-eosin. The pathological changes of the skin were observed under a microscope and photographed. The epidermal thickness was calculated using the pathological image analysis system Image-Pro plus.

[0261] 6. Results and analysis

[0262] Animal experiments showed that, under the same dosage conditions, compound 2 could reduce the PASI score of imiquimod-induced psoriasis in mice, improve the clinical pathological appearance of psoriasis mice, and had a good inhibitory effect on abnormal thickening of the epidermis in psoriasis mice. The pharmacodynamic effect on psoriasis mice was: compound 2> baicalein (YHQ), as shown in Table 7 for details.

[0263] Table 7 Evaluation of the efficacy of compound 2 on psoriasis in mice

[0264]

[0265] Example 11: Inhibitory effect of compound 2 on proliferation of human hepatic stellate cells LX-2

[0266] 1. Methods

[0267] 1.1 LX-2 cell culture

[0268] LX-2 cells (provided by Beijing Beina Chuanglian Biotechnology Research Institute) were cultured in culture bottles containing RPMI1640 culture medium provided by (Wuhan Pronosai Life Science Technology Co., Ltd.), the culture medium contained 10% fetal bovine serum (Zhejiang Tianhang Biotechnology Co., Ltd.), 100 mg / mL streptomycin, and 100 kU / L penicillin, and were placed in a 5% CO2 incubator (Binder, Germany) at saturated humidity and 37°C.

[0269] 1.2. MTT assay to detect cell proliferation activity

[0270] LX-2 cells in the logarithmic growth phase were taken and digested with 0.25% trypsin. The cell density was adjusted to 5×10 cells using RPMI1640 medium (10% FBS + 1% double antibody). 4 / mL, access to 96-well plate. The cells were divided into a normal control group (no drug), a TGF-β1 (TGF-β1 was added to the well plate at a concentration of 10ng / mL) group, a scutellarin (YHQ) (YHQ was added to the well plate at a concentration of 10μM) group, and an Example 2 compound 2 (Compound 2 prepared in Example 2 was added to the well plate at a concentration of 10μM) group. Each group had 6 duplicate wells, 100μL of cell suspension per well, and a blank zeroing well was set. 200μL of sterile PBS buffer was added around the cell wells. After culturing for 24h at 37°C and 5% CO2 saturated humidity, 20μL of MTT was added to each well, and the same culture conditions were used for 3h. The culture medium was aspirated, and 100μL of DMSO was added for oscillation for 10min. The absorbance (A) at 490nm of each well was measured by an ELISA instrument to calculate the cell proliferation rate. Cell proliferation rate = (A experimental group - A zero-adjustment group) / (A control group - A zero-adjustment group) × 100%.

[0271] The experimental results are shown in Table 8. It can be seen from Table 8 that compound 2 prepared in Example 2 has a good inhibitory effect on the proliferation of LX-2 cells.

[0272] Table 8 Inhibition data of different drugs on LX-2 cells

[0273] sample Normal group TGF-β1 group Example 2 Compound 2 YHQ Group Cell survival rate / % 100 120 50 75

[0274] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

[0275] The present invention provides a transient receptor potential cation channel TRPV3 inhibitor and its use ideas and methods. There are many methods and ways to implement the technical solution. The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, without departing from the principle of the present invention, several improvements and modifications can be made, and these improvements and modifications should also be regarded as the protection scope of the present invention. All components not specified in this embodiment can be implemented by existing technologies.

Claims

1. A compound of formula I or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs; in, R1 is independently selected from hydrogen, deuterium, substituted or unsubstituted amino, hydroxyl, thiol, cyano, nitro, substituted or unsubstituted C 1-6 Alkyl, substituted or unsubstituted C 1-6 Alkoxy, substituted or unsubstituted C 1-6 Haloalkyl, substituted or unsubstituted C 1-6 Haloalkoxy, substituted or unsubstituted C 1-6 Alkylamino, carboxyl, substituted or unsubstituted C 1-6 Alkanoyl, substituted or unsubstituted C 1-6 alkylsulfonyl, substituted or unsubstituted aminoacyl, or substituted or unsubstituted aminosulfonyl; wherein the substitution is selected from hydrogen, deuterium, halogen, hydroxyl or C 1-6 Preferably, R1 is independently selected from hydrogen, deuterium, substituted or unsubstituted amino, hydroxyl, thiol, cyano, nitro, substituted or unsubstituted C 1-3 Alkyl, substituted or unsubstituted C 1-3 Alkoxy, substituted or unsubstituted C 1-3 Haloalkyl, substituted or unsubstituted C 1-3 Haloalkoxy, substituted or unsubstituted C 1-3 Alkylamino, carboxyl, substituted or unsubstituted C 1-3 Alkanoyl, substituted or unsubstituted C 1-3 alkylsulfonyl, substituted or unsubstituted aminoacyl, or substituted or unsubstituted aminosulfonyl; wherein the substitution is selected from hydrogen, deuterium, halogen, hydroxyl or C 1-3 Alkyl substitution; R2 is independently selected from hydrogen, deuterium, substituted or unsubstituted amino, hydroxyl, thiol, cyano, nitro, halogen, substituted or unsubstituted C 1-6 Alkyl, substituted or unsubstituted C 1-6 Alkoxy, substituted or unsubstituted C 1-6 Haloalkyl, substituted or unsubstituted C 1-6 Haloalkoxy, substituted or unsubstituted C 1-6 Alkylamino, carboxyl, substituted or unsubstituted C 1-6 Alkanoyl, substituted or unsubstituted C 1-6 alkylsulfonyl, substituted or unsubstituted aminoacyl, or substituted or unsubstituted aminosulfonyl; wherein the substitution is selected from hydrogen, deuterium, halogen, hydroxyl or C 1-6 Preferably, R2 is independently selected from hydrogen, deuterium, substituted or unsubstituted amino, hydroxyl, thiol, cyano, nitro, halogen, substituted or unsubstituted C 1-3 Alkyl, substituted or unsubstituted C 1-3 Alkoxy, substituted or unsubstituted C 1-3 Haloalkyl, substituted or unsubstituted C 1-3 Haloalkoxy, substituted or unsubstituted C 1-3 Alkylamino, carboxyl, substituted or unsubstituted C 1-3 Alkanoyl, substituted or unsubstituted C 1-3 alkylsulfonyl, substituted or unsubstituted aminoacyl, or substituted or unsubstituted aminosulfonyl; wherein the substitution is selected from hydrogen, deuterium, halogen, hydroxyl or C 1-3 Alkyl substitution; n is an integer selected from 1-4, preferably an integer selected from 1-3; m is an integer selected from 0-3, preferably an integer selected from 0-2; Virtual bonds are represented as non-existent, single bonds, or double bonds, and virtual bonds are not always double bonds at the same time; When the virtual bond is represented as absent, X is selected from -NR3R4, -CR5R6R7, -OR8 or -SR9, preferably selected from -NR3R4 or -OR8; When the virtual bond connected to X is represented as a single bond, X is selected from -NR3-, -CR5R6-, -O- or -S-, preferably -NR3- or -O-; R3 is independently selected from hydrogen, deuterium, substituted or unsubstituted amino, hydroxyl, thiol, cyano, nitro, substituted or unsubstituted C 1-6 Alkyl, substituted or unsubstituted C 1-6 Alkoxy, substituted or unsubstituted C 1-6 Haloalkyl, substituted or unsubstituted C 1-6 Haloalkoxy, substituted or unsubstituted C 1-6 Alkylamino, carboxyl, substituted or unsubstituted C 1-6 Alkanoyl, substituted or unsubstituted C 1-6 alkylsulfonyl, substituted or unsubstituted aminoacyl, or substituted or unsubstituted aminosulfonyl; wherein the substitution is selected from hydrogen, deuterium, halogen, hydroxyl or C 1-6 Preferably, R3 is independently selected from hydrogen, deuterium, substituted or unsubstituted amino, hydroxyl, thiol, cyano, nitro, substituted or unsubstituted C 1-3 Alkyl, substituted or unsubstituted C 1-3 Alkoxy, substituted or unsubstituted C 1-3 Haloalkyl, substituted or unsubstituted C 1-3 Haloalkoxy, substituted or unsubstituted C 1-3 Alkylamino, carboxyl, substituted or unsubstituted C 1-3 Alkanoyl, substituted or unsubstituted C 1-3 alkylsulfonyl, substituted or unsubstituted aminoacyl, or substituted or unsubstituted aminosulfonyl; wherein the substitution is selected from hydrogen, deuterium, halogen, hydroxyl or C 1-3 Alkyl substitution; R4 is independently selected from hydrogen, deuterium, substituted or unsubstituted amino, hydroxyl, thiol, cyano, nitro, substituted or unsubstituted C 1-6 Alkyl, substituted or unsubstituted C 1-6 Alkoxy, substituted or unsubstituted C 1-6 Haloalkyl, substituted or unsubstituted C 1-6 Haloalkoxy, substituted or unsubstituted C 1-6 Alkylamino, carboxyl, substituted or unsubstituted C 1-6 Alkanoyl, substituted or unsubstituted benzoyl, substituted or unsubstituted C 1-6 alkylsulfonyl, substituted or unsubstituted aminoacyl, or substituted or unsubstituted aminosulfonyl; wherein the substitution is selected from hydrogen, deuterium, halogen, hydroxyl and C 1-6 Preferably, R4 is independently selected from hydrogen, deuterium, substituted or unsubstituted amino, hydroxyl, thiol, cyano, nitro, substituted or unsubstituted C 1-3 Alkyl, substituted or unsubstituted C 1-3 Alkoxy, substituted or unsubstituted C 1-3 Haloalkyl, substituted or unsubstituted C 1-3 Haloalkoxy, substituted or unsubstituted C 1-3 Alkylamino, carboxyl, substituted or unsubstituted C 1-3 Alkanoyl, substituted or unsubstituted benzoyl, substituted or unsubstituted C 1-3 alkylsulfonyl, substituted or unsubstituted aminoacyl, or substituted or unsubstituted aminosulfonyl; wherein the substitution is selected from hydrogen, deuterium, halogen, hydroxyl and C 1-3 The alkyl group may be substituted by one or more identical or different groups; R5, R6, R7, R8, and R9 are independently selected from hydrogen, deuterium, substituted or unsubstituted amino, hydroxyl, thiol, cyano, nitro, substituted or unsubstituted C 1-6 Alkyl, substituted or unsubstituted C 1-6 Alkoxy, substituted or unsubstituted C 1-6 Haloalkyl, substituted or unsubstituted C 1-6 Haloalkoxy, substituted or unsubstituted C 1-6 Alkylamino, carboxyl, substituted or unsubstituted C 1-6 Alkanoyl, substituted or unsubstituted C 1-6 alkylsulfonyl, substituted or unsubstituted aminoacyl, substituted or unsubstituted aminosulfonyl, substituted or unsubstituted C 3-8 Cycloalkyl, substituted or unsubstituted C 6-10 aryl, substituted or unsubstituted heterocyclic group consisting of 3-8 atoms or substituted or unsubstituted heteroaryl consisting of 5-10 atoms; preferably, R5, R6, R7, R8, R9 are independently selected from hydrogen, deuterium, substituted or unsubstituted amino, hydroxyl, thiol, cyano, nitro, substituted or unsubstituted C 1-3 Alkyl, substituted or unsubstituted C 1-3 Alkoxy, substituted or unsubstituted C 1-3 Haloalkyl, substituted or unsubstituted C 1-3 Haloalkoxy, substituted or unsubstituted C 1-3 Alkylamino, carboxyl, substituted or unsubstituted C 1-3 Alkanoyl, substituted or unsubstituted C 1-3 alkylsulfonyl, substituted or unsubstituted aminoacyl, substituted or unsubstituted aminosulfonyl, substituted or unsubstituted C 3-5 Cycloalkyl, substituted or unsubstituted C 6-8 Aryl, substituted or unsubstituted heterocyclic group consisting of 3 to 8 atoms or substituted or unsubstituted heteroaryl group consisting of 5 to 10 atoms.

2. The compound of formula (I) according to any one of claim 1 or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs, characterized in that: The compound of formula I is shown in formula I-1 or formula I-2: In formula I-1, R1 is independently selected from hydrogen, deuterium, hydroxyl or substituted or unsubstituted C 1-3 wherein the substitution is selected from hydrogen, deuterium, hydroxyl or C 1-3 Alkyl substitution; preferably, R1 is independently selected from hydrogen, deuterium, hydroxyl or substituted or unsubstituted ethyl; wherein the substitution is selected from methyl substitution; R2 is independently selected from substituted or unsubstituted C 1-3 Alkyl, hydrogen, deuterium, hydroxyl or halogen; the substitution is selected from hydrogen, deuterium, halogen, hydroxyl or C 1-3 Alkyl substituted; preferably, R2 is independently selected from hydrogen, deuterium, hydroxyl, isopropyl, fluorine or chlorine; n is further preferably selected from 2 or 3; m is further preferably selected from 1 or 2; X is preferably selected from -NR3- or -O-; R3 is independently selected from hydrogen or deuterium; In formula I-2, R1 is independently selected from hydrogen, deuterium or hydroxyl; n is further preferably selected from 2; X is further preferably selected from -NR3R4; R3 is independently selected from hydrogen, deuterium or C 1-3 Alkyl; preferably, R3 is independently selected from hydrogen or deuterium; R4 is independently selected from hydrogen, deuterium or substituted or unsubstituted benzoyl; wherein the substitution is selected from substitution with one or more identical or different groups selected from hydrogen, deuterium, halogen and hydroxyl; preferably, R4 is independently selected from substituted or unsubstituted benzoyl; wherein the substitution is selected from substitution with one or more identical or different groups selected from hydrogen, deuterium, fluorine, chlorine and hydroxyl.

3. The compound of formula (I) according to claim 2 or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs, characterized in that: The compound of formula I is shown in formula I-1-1, formula I-1-2 or formula I-1-3: In formula I-1-1, R 10 independently selected from hydrogen, deuterium, hydroxyl or substituted or unsubstituted ethyl; wherein the substitution is selected from hydrogen, deuterium, hydroxyl or C 1-3 Alkyl substituted; preferably, R 10 are independently selected from hydrogen, deuterium, hydroxyl or substituted or unsubstituted ethyl; wherein the substitution is selected from C 1-3 Alkyl substitution; R 11 , R 12 independently selected from hydrogen, deuterium, hydroxyl; R2 is independently selected from hydrogen, deuterium, hydroxyl; In formula I-1-2, R1 is independently selected from hydroxyl; R3 is independently selected from hydrogen or deuterium; R 13 independently selected from substituted or unsubstituted ethyl; wherein the substitution is selected from hydrogen, deuterium, hydroxyl or C 1-3 Alkyl substituted; preferably, R 13 independently selected from substituted or unsubstituted ethyl; wherein the substitution is selected from C 1-3 Alkyl substitution; In formula I-1-3, R1 is independently selected from hydroxyl; R3 is independently selected from hydrogen or deuterium; R 14 , R 15 are independently selected from hydroxyl or halogen; preferably, R 14 , R 15 are independently selected from hydroxy, fluoro or chloro.

4. The compound of formula (I) according to claim 1 or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs, characterized in that: It is selected from any of the following compounds:

5. The compound according to any one of claims 1 to 4, characterized in that Prepared by the following method, select one of the following synthetic routes: Synthetic route 1: wherein R2 is selected from substituted or unsubstituted ethyl, hydroxyl or halogen; wherein the substitution is selected from hydrogen, deuterium, hydroxyl or C 1-3 Alkyl substituted; m is selected from 1 or 2; or, Synthesis route 2: Among them, R 16 is selected from hydrogen, deuterium, halogen or hydroxyl; z is selected from 2; or, Synthetic route 3:

6. A method for preparing a compound according to any one of claims 1 to 4, characterized in that: Choose one of the following synthetic routes: Synthetic route 1: wherein R2 is selected from substituted or unsubstituted ethyl, hydroxyl or halogen; wherein the substitution is selected from hydrogen, deuterium, hydroxyl or C 1-3 Alkyl substituted; m is selected from 1 or 2; or, Synthesis route 2: Among them, R 16 is selected from hydrogen, deuterium, halogen or hydroxyl; z is selected from 2; or, Synthetic route 3:

7. A pharmaceutical composition, characterized in that Comprising the compound of formula I according to claim 1 or any one of its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs, and one or more pharmaceutically acceptable carriers; preferably, the pharmaceutical composition is in the form of a subcutaneous injection, intradermal injection, spray, powder aerosol, external solution, lotion, liniment, ointment, plaster, paste, patch or one or more thereof.

8. A pharmaceutical external ointment containing the compound according to claim 1 as an active ingredient, characterized in that: Including oil phase, water phase and active ingredients, the weight percentage of each material is: Oil phase: Light liquid paraffin 0.6-3%, preferably 0.8-2%, more preferably 1.00%; Hexadecanol 1.5-7.5%, preferably 2.0-5.0%, more preferably 2.54%; Stearic acid 2.6-5.0%, preferably 3.2-4.4%, more preferably 3.80%; Aqueous phase: Glycerol 5.4-27%, preferably 7.2-18%, more preferably 8.92%; Triethylamine alcohol 0.27-1.38%, preferably 0.35-0.92%, more preferably 0.46%; Butylparaben 0.06-3%, preferably 0.08-2%, more preferably 0.10%; Active ingredient 0.6-12%, preferably 0.8-8%, more preferably 1% to 4%; The balance is purified water; The active ingredient is the compound of formula I according to claim 1 or its stereoisomer, geometric isomer, tautomer, nitrogen oxide, hydrate, solvate, pharmaceutically acceptable salt or prodrug; Preferably, the method for preparing the pharmaceutical external ointment comprises the following steps: (1) dissolving the active ingredient in part of purified water to obtain a drug solution; mixing light liquid paraffin, hexyl alcohol, and stearic acid and heating and melting them to obtain an oil phase; mixing glycerin, triethylamine alcohol, butyl hydroxybenzoate, and the remaining purified water and heating and melting them to obtain a water phase; (2) adding the drug solution obtained in step (1) to the aqueous phase and mixing to obtain an aqueous phase containing the drug; adding the aqueous phase containing the drug to the oil phase and stirring until the temperature cools to room temperature.

9. Use of the compound of formula I according to claim 1 or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs, or the pharmaceutical composition according to claim 7, or the pharmaceutical external ointment according to claim 8 in the preparation of a TRPV3 inhibitor, or in the preparation of a drug for treating itching, atopic dermatitis, and pain caused by kidney disease, or in the preparation of a drug for treating kidney disease, or in the preparation of a drug for treating anti-fibrosis.

10. Use of the compound of formula I according to claim 1 or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs, or the pharmaceutical composition according to claim 7, or the pharmaceutical external ointment according to claim 8 in the preparation of a medicament for treating a TRPV3-mediated disease; preferably, the treatment of a TRPV3-mediated disease is the treatment of a TRPV3-mediated disease by inhibiting the increase in calcium ion influx caused by TRPV3 activation; further preferably, the TRPV3-mediated disease is one or more of pruritus, alopecia, atopic dermatitis, psoriasis, and ulcerative colitis, and further preferably, the TRPV3-mediated disease is psoriasis; the treatment of TRPV3-mediated psoriasis is the improvement of psoriasis-like lesions, psoriatic arthritis or psoriasis pain.

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