Transient receptor potential cation channel, subfamily v, member 3 (trpv3) inhibitors and uses thereof

Abstract

This invention belongs to the field of pharmaceutical technology, specifically relating to a transient receptor potential cation channel (TRPV3) inhibitor and its uses. The compound provided by this invention can act as a TRPV3 inhibitor, suppressing the abnormal increase in calcium ion influx caused by TRPV3 activation. It can effectively treat and prevent TRPV3-related conditions, providing new approaches for conditions such as itching, hair loss, atopic dermatitis, psoriasis, and ulcerative colitis. The compound provided by this invention exhibits better solubility while maintaining a comparable inhibition rate of calcium ion influx. Furthermore, the compound of this invention shows an inhibitory effect on the proliferation of human hepatic stellate cells (LX-2), demonstrating anti-fibrotic activity.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, specifically relating to a quinolinone compound with TRPV3 inhibitory activity and its pharmaceutical composition, and further relating to its preparation method and pharmaceutical use. Background Technology

[0002] TRPV3 is a non-selective cation channel with relatively high calcium ion permeability. First cloned in 2002, it shows 30%–40% sequence homology with other TRPV channels. The TRPV3 channel is a homotetramer formed by six transmembrane subunits containing multiple repeating ankyrin domains (ARD), N-terminal and C-terminal, with pores forming loops between the domains.

[0003] As a member of the temperature-sensitive TRP family, the non-selective cation channel TRPV3, which osmotically penetrates calcium ions, is highly expressed in dermal keratinocytes. Weak acids can diffuse onto the cell membrane in a protonated form, subsequently releasing free protons, leading to intracellular acidification and TRPV3 activation. Protons can also osmotically penetrate activated TRPV3 or other proton channels, causing intracellular acidification and TRPV3 activation. TRPV3 activation mediates calcium influx and induces calcium cytoplasmic overload, leading to keratinization and cell death or dermatopathy, accompanied by severe mast cell infiltration and histamine release in the upper dermis; furthermore, TRPV3-mediated nitric oxide release in keratinocytes promotes wound healing and causes pain. Recently, gain-of-function mutations in human TRPV3, characterized by severe pruritus and palmoplantar and perioral keratosis, were discovered in patients with Olmsted syndrome, revealing its crucial role in chronic dermatopathy. In mice, loss-of-function mutations in the transforming growth factor α (TGF-α) and epidermal growth factor receptor (EGFR) genes induce a wavy hair phenotype, while upregulation of TGF-α / EGFR signaling results in a hairless phenotype. Furthermore, small interfering RNA-mediated silencing of TRPV3 effectively eliminates TRPV3 agonist-induced cellular effects, including functional currents, reduced proliferation, increased intracellular calcium concentration, and apoptosis.

[0004] Currently, many scientists have begun to focus on the discovery of TRPV3 inhibitors. TRPV3 forms a signaling complex with TGF-α / EGFR. Through the Ca2+ pathway of TRPV3... 2+Influx of EGFRs leads to the release / shedding of TGF-α, which in turn activates the EGFR-mediated signaling pathway that promotes keratinocyte proliferation. The pathogenesis of psoriasis is related to the excessive proliferation of immune cells and keratinocytes and the massive release of inflammatory factors following the immune response. Simultaneously, EGFR activation leads to increased TRPV3 channel activity, thereby stimulating TGF-α release. In conclusion, selectively targeting and inhibiting TRPV3 may be an effective method for treating various skin diseases such as keratosis, alopecia, pruritus, and psoriasis.

[0005] Glenmark has filed patents for a series of TRPV3 antagonists, including US20100292254, US20090286811, and US20100311778. In 2010, Glenmark entered into a licensing agreement with Sanofi-Aventis, subsequently pushing its lead molecule (GRC15300, structure unknown) into clinical trials for the treatment of osteoarthritis and neuropathic pain. In 2012, GRC15300 entered a Phase II trial for the treatment of neuropathic pain; however, these trials were discontinued by the end of 2013. The small molecule structure of the TRPV3 inhibitor 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 baicalein as an allosteric negative regulator of (Scutellarein, Scu) TRPV3 channels with an epigenetic affinity of 1.18 μM. It can attenuate endogenous TRPV3 activity in human keratinocytes and inhibit carvacrol-induced proliferation and pro-inflammatory responses. The structural formula of baicalein is as follows:

[0008] Summary of the Invention

[0009] The technical problem to be solved by the present invention is to address the shortcomings of the prior art by providing a quinolone compound and its pharmaceutical composition as a TRPV3 inhibitor, which can be used to treat various inflammatory skin diseases, such as psoriasis.

[0010] To solve the above-mentioned technical problems, the technical solution adopted by the present invention is as follows:

[0011] This invention discloses a compound of Formula I or a stereoisomer, geometric isomer, tautomer, nitride, hydrate, solvate, pharmaceutically acceptable salt or prodrug thereof;

[0012]

[0013] in,

[0014] R1 is independently selected from hydrogen, deuterium, substituted or unsubstituted amino, hydroxyl, mercapto, cyano, nitro, substituted or unsubstituted C. 1-6 Alkyl, substituted or unsubstituted C 1-6 Alkoxy, substituted or unsubstituted C 1-6 Halogenated alkyl, substituted or unsubstituted C 1-6 Halogenated alkoxy, substituted or unsubstituted C 1-6 Alkylamino, carboxyl, substituted or unsubstituted C 1-6 Alkyl group, substituted or unsubstituted C 1-6 Alkyl sulfonyl, 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, mercapto, cyano, nitro, halogen, substituted or unsubstituted C. 1-6 Alkyl, substituted or unsubstituted C 1-6 Alkoxy, substituted or unsubstituted C 1-6 Halogenated alkyl, substituted or unsubstituted C 1-6 Halogenated alkoxy, substituted or unsubstituted C 1-6 Alkylamino, carboxyl, substituted or unsubstituted C 1-6 Alkyl group, substituted or unsubstituted C 1-6 Alkyl sulfonyl, 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 selected from integers between 0 and 3;

[0018] A virtual key represents a non-existent key, a single key, or a double key, and a virtual key cannot be a double key at the same time;

[0019] When a virtual key is indicated as not existing, 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, mercapto, cyano, nitro, substituted or unsubstituted C. 1-6 Alkyl, substituted or unsubstituted C 1-6 Alkoxy, substituted or unsubstituted C 1-6 Halogenated alkyl, substituted or unsubstituted C 1-6 Halogenated alkoxy, substituted or unsubstituted C 1-6 Alkylamino, carboxyl, substituted or unsubstituted C 1-6 Alkyl group, substituted or unsubstituted C 1-6 Alkyl sulfonyl, 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, mercapto, cyano, nitro, substituted or unsubstituted C. 1-6 Alkyl, substituted or unsubstituted C 1-6 Alkoxy, substituted or unsubstituted C 1-6 Halogenated alkyl, substituted or unsubstituted C 1-6 Halogenated alkoxy, substituted or unsubstituted C 1-6 Alkylamino, carboxyl, substituted or unsubstituted C 1-6 Alkyl group, substituted or unsubstituted benzoyl group, substituted or unsubstituted C 1-6 Alkyl sulfonyl, substituted or unsubstituted amino acyl, or substituted or unsubstituted aminosulfonyl; wherein the substitution is selected from hydrogen, deuterium, halogen, hydroxyl, and C. 1-6 One or more identical or different groups are substituted in an alkyl group;

[0023] R5, R6, R7, R8, and R9 are independently selected from hydrogen, deuterium, substituted or unsubstituted amino groups, hydroxyl groups, mercapto groups, cyano groups, nitro groups, and substituted or unsubstituted C groups. 1-6 Alkyl, substituted or unsubstituted C 1-6 Alkoxy, substituted or unsubstituted C 1-6 Halogenated alkyl, substituted or unsubstituted C 1-6 Halogenated alkoxy, substituted or unsubstituted C 1-6 Alkylamino, carboxyl, substituted or unsubstituted C 1-6 Alkyl group, substituted or unsubstituted C 1-6Alkyl sulfonyl, substituted or unsubstituted amino acyl, substituted or unsubstituted aminosulfonyl, substituted or unsubstituted C 3-8 Cycloalkyl, substituted or unsubstituted C 6-10 Aryl, heterocyclic group consisting of 3-8 atoms (substituted or unsubstituted), or heteroaryl group consisting of 5-10 atoms (substituted or unsubstituted).

[0024] In some embodiments, preferably, R1 is independently selected from hydrogen, deuterium, substituted or unsubstituted amino, hydroxyl, mercapto, cyano, nitro, substituted or unsubstituted C. 1-3 Alkyl, substituted or unsubstituted C 1-3 Alkoxy, substituted or unsubstituted C 1-3 Halogenated alkyl, substituted or unsubstituted C 1-3 Halogenated alkoxy, substituted or unsubstituted C 1-3 Alkylamino, carboxyl, substituted or unsubstituted C 1-3 Alkyl group, substituted or unsubstituted C 1-3 Alkyl sulfonyl, 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, mercapto, cyano, nitro, halogen, substituted or unsubstituted C. 1-3 Alkyl, substituted or unsubstituted C 1-3 Alkoxy, substituted or unsubstituted C 1-3 Halogenated alkyl, substituted or unsubstituted C 1-3 Halogenated alkoxy, substituted or unsubstituted C 1-3 Alkylamino, carboxyl, substituted or unsubstituted C 1-3 Alkyl group, substituted or unsubstituted C 1-3 Alkyl sulfonyl, 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 an integer selected from 0 to 2;

[0028] A virtual key represents a non-existent key, a single key, or a double key, and a virtual key cannot be a double key at the same time;

[0029] When the virtual key is indicated as not existing, 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, mercapto, cyano, nitro, substituted or unsubstituted C. 1-3 Alkyl, substituted or unsubstituted C 1-3 Alkoxy, substituted or unsubstituted C 1-3 Halogenated alkyl, substituted or unsubstituted C 1-3 Halogenated alkoxy, substituted or unsubstituted C 1-3 Alkylamino, carboxyl, substituted or unsubstituted C 1-3 Alkyl group, substituted or unsubstituted C 1-3 Alkyl sulfonyl, 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, mercapto, cyano, nitro, substituted or unsubstituted C. 1-3 Alkyl, substituted or unsubstituted C 1-3 Alkoxy, substituted or unsubstituted C 1-3 Halogenated alkyl, substituted or unsubstituted C 1-3 Halogenated alkoxy, substituted or unsubstituted C 1-3 Alkylamino, carboxyl, substituted or unsubstituted C 1-3 Alkyl group, substituted or unsubstituted benzoyl group, substituted or unsubstituted C 1-3 Alkyl sulfonyl, substituted or unsubstituted amino acyl, or substituted or unsubstituted aminosulfonyl; wherein the substitution is selected from hydrogen, deuterium, halogen, hydroxyl, and C. 1-3 One or more identical or different groups are substituted in an alkyl group;

[0033] R5, R6, R7, R8, and R9 are independently selected from hydrogen, deuterium, substituted or unsubstituted amino groups, hydroxyl groups, mercapto groups, cyano groups, nitro groups, and substituted or unsubstituted C groups. 1-3 Alkyl, substituted or unsubstituted C 1-3 Alkoxy, substituted or unsubstituted C 1-3 Halogenated alkyl, substituted or unsubstituted C 1-3 Halogenated alkoxy, substituted or unsubstituted C 1-3 Alkylamino, carboxyl, substituted or unsubstituted C 1-3 Alkyl group, substituted or unsubstituted C 1-3 Alkyl sulfonyl, substituted or unsubstituted amino acyl, substituted or unsubstituted aminosulfonyl, substituted or unsubstituted C 3-5Cycloalkyl, substituted or unsubstituted C 6-8 Aryl, heterocyclic group consisting of 3-8 atoms (substituted or unsubstituted), or heteroaryl group consisting of 5-10 atoms (substituted or unsubstituted).

[0034] In some embodiments, and more preferably, the compound represented by Formula I above, or a stereoisomer, geometric isomer, tautomer, nitride, hydrate, solvate, pharmaceutically acceptable salt, or prodrug thereof,

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

[0036]

[0037] in,

[0038] R1 is independently selected from hydrogen, deuterium, hydroxyl, or substituted or unsubstituted C. 1-3 Alkyl; 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 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, and more preferably, R1 is independently selected from hydrogen, deuterium, hydroxyl, or substituted or unsubstituted ethyl groups; wherein the substitution is selected from those substituted with methyl groups;

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

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

[0047] In some embodiments, and more preferably, the compound represented by Formula I above, or a stereoisomer, geometric isomer, tautomer, nitride, hydrate, solvate, pharmaceutically acceptable salt, or prodrug thereof,

[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 groups; wherein the substitution is selected from those substituted with hydrogen, deuterium, hydroxyl, or C. 1-3 Alkyl substitution;

[0051] R 11 R 12 Independently selected from hydrogen, deuterium, and hydroxyl groups;

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

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

[0054] In some embodiments, and more preferably, the compound represented by Formula I above, or a stereoisomer, geometric isomer, tautomer, nitride, hydrate, solvate, pharmaceutically acceptable salt, or prodrug thereof,

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

[0056]

[0057] R1 is independently selected from hydroxyl groups;

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

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

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

[0061] In some embodiments, and more preferably, the compound represented by Formula I above, or a stereoisomer, geometric isomer, tautomer, nitride, hydrate, solvate, pharmaceutically acceptable salt, or prodrug thereof,

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

[0063]

[0064] R1 is independently selected from hydroxyl groups;

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

[0066] R 14 R 15 It is independently selected from hydroxyl or halogen.

[0067] Specifically, more preferably, R 14 R 15 It is independently selected from hydroxyl, fluorine or chlorine.

[0068] In some embodiments, and more preferably, the compound represented by Formula I above, or a stereoisomer, geometric isomer, tautomer, nitride, hydrate, solvate, pharmaceutically acceptable salt, or prodrug thereof,

[0069] When all dummy bonds are represented 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 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 groups; wherein the substitution is selected from one or more identical or different groups selected from hydrogen, deuterium, halogen, and hydroxyl.

[0076] Specifically, and 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 groups; wherein the substitution is selected from one or more of the same or different groups selected from hydrogen, deuterium, fluorine, chlorine and hydroxyl.

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

[0080]

[0081] The compound represented by Formula I above, or its stereoisomers, geometric isomers, tautomers, nitrides, hydrates, solvates, pharmaceutically acceptable salts, or prodrugs, are prepared by the following methods, choosing one of the following synthetic routes:

[0082] Synthesis 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 substitution; m is selected from 1 or 2;

[0085] or,

[0086] Synthesis Route 2:

[0087]

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

[0089] or,

[0090] Synthesis Route 3:

[0091]

[0092] Furthermore, this invention discloses a method for preparing the compound represented by Formula I above, or its stereoisomers, geometric isomers, tautomers, nitrides, hydrates, solvates, pharmaceutically acceptable salts, or prodrugs, by selecting one of the following synthetic routes:

[0093] Synthesis Route 1:

[0094] 2-Amino-4,5-dimethoxyacetophenone reacts with compound a-1 under the action of a first base to undergo a first acylation reaction to give intermediate b-1; intermediate b-1 undergoes a first intramolecular cyclization reaction under the action of a second base to give intermediate c-1; intermediate c-1 undergoes a first demethylation reaction under the action of a first Lewis acid to obtain compound IA.

[0095]

[0096] 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 substitution; m is selected from 1 or 2;

[0097] or,

[0098] Synthesis Route 2:

[0099] 2-Amino-4,5-dimethoxyacetophenone reacts with compound a-2 under the action of a third base to undergo a second acylation reaction, yielding intermediate b-2; intermediate b-2 then undergoes a second demethylation reaction under the action of a second Lewis acid, thus producing compound IB.

[0100]

[0101] Among them, R 16 Selected from hydrogen, deuterium, halogen, or hydroxyl; z is selected from 2;

[0102] or,

[0103] Synthesis Route 3:

[0104] 3,5-Dimethoxyphenol reacts with 2-chloropropane in the presence of a first catalyst to undergo a first substitution reaction, yielding compound a-3; compound a-3 reacts with acetyl chloride in the presence of a second catalyst to undergo a second substitution reaction, yielding compound b-3; compound b-3 reacts with 4-methoxybenzoyl chloride in the presence of a fourth base to undergo an esterification rearrangement reaction, yielding compound c-3; compound c-3 undergoes a cyclization reaction in the presence of an inorganic acid, yielding compound d-3; compound d-3 undergoes a third demethylation reaction in the presence of a third Lewis acid, yielding compound 1.

[0105]

[0106] Specifically, in synthetic route 1, the first base is triethylamine, ethylenediamine, or n-propylamine, preferably triethylamine; the molar ratio of 2-amino-4,5-dimethoxyacetophenone to compound a-1 and the first base is 1:0.9–4.5:2–70:, preferably 1:1.2–3:2.4–14, more preferably 1:1.5:2.8; the solvent used in the first acylation reaction is tetrahydrofuran, methyltetrahydrofuran, or dimethyltetrahydrofuran, preferably tetrahydrofuran; the first The acylation reaction is carried out at a temperature of 45–85°C, preferably 55–75°C, more preferably 65°C, for a reaction time of 2.4–12 h, preferably 3.2–8 h, more preferably 4 h; the second base is sodium hydroxide, potassium hydroxide, or calcium hydroxide, preferably sodium hydroxide; the molar ratio of intermediate b-1 to the second base is 1:4–26, preferably 1:5–13, more preferably 1:6–7.2, and even more preferably 1:6.6; the first intramolecular cyclization reaction… The solvent used in the process is toluene, xylene, or 1,4-dioxane, preferably 1,4-dioxane; the first intramolecular cyclization reaction is carried out at a temperature of 90–130°C, preferably 100–120°C, more preferably 110°C, for a reaction time of 2.4–12 h, preferably 3.2–8 h, more preferably 4 h; the first Lewis acid is boron tribromide, boron trichloride, or aluminum trichloride, preferably boron tribromide; the molar ratio of intermediate c-1 to the first Lewis acid is 1:4–26, preferably 1:5–13, more preferably 1:6–6.5, and even more preferably 1:6.25; the first Lewis acid exists in solution form, the solvent is dichloromethane, and the concentration of the first Lewis acid in the solution is 1.2–6 mol / L, preferably 1.6–4 mol / L, more preferably 2 mol / L; the first demethylation reaction is carried out at room temperature for a reaction time of 2.4–12 h, preferably 3.2–8 h, more preferably 4 h.

[0107] 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.

[0108] Specifically, in synthetic route 2, the third base is triethylamine, ethylenediamine, or n-propylamine, preferably triethylamine; the molar ratio of 2-amino-4,5-dimethoxyacetophenone to compound a-2 and the third base is 1:0.9–4.5:2–70:, preferably 1:1.2–3:2.4–14, more preferably 1:1.5:2.8; the solvent used in the second acylation reaction is tetrahydrofuran, methyltetrahydrofuran, or dimethyltetrahydrofuran, preferably tetrahydrofuran; the reaction temperature of the second acylation reaction is 45–85°C, preferably 55–75°C, more preferably 65°C, and the reaction time is 2.4–12 h, preferably 3.2–8 h. The preferred step is 4 hours; the second Lewis acid is boron tribromide, boron trichloride, or aluminum trichloride, preferably boron tribromide; the molar ratio of intermediate b-2 to the second Lewis acid is 1:4 to 26, preferably 1:5 to 13, more preferably 1:6 to 6.5, and even more preferably 1:6.25; the second Lewis acid exists in solution form, the solvent is dichloromethane, and the concentration of the second Lewis acid in the solution is 1.2 to 6 mol / L, preferably 1.6 to 4 mol / L, and more preferably 2 mol / L; the second demethylation reaction is carried out at room temperature for 2.4 to 12 hours, preferably 3.2 to 8 hours, and more preferably 4 hours.

[0109] Specifically, in synthetic route 2, the second acylation reaction is preferably carried out under inert gas protection; the second demethylation reaction is preferably carried out under inert gas protection; wherein, the inert gas is preferably nitrogen.

[0110] Specifically, in synthetic route 3, the first catalyst is boron tribromide, boron trichloride, or aluminum trichloride, preferably aluminum trichloride; the molar ratio of 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 for the first substitution reaction is dichloromethane or trichloromethane, preferably trichloromethane; the reaction temperature for the first substitution reaction is room temperature, and the reaction time is 2.4–12 h, preferably 3.2–8 h, and more preferably 4 h; the second catalyst is boron tribromide, boron trichloride, or aluminum trichloride, preferably... The first base is aluminum trichloride; the molar ratio of 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, more preferably 1:1.2:2; the solvent used for the second substitution reaction is dichloromethane or trichloromethane, preferably trichloromethane; the second substitution reaction is carried out at room temperature for 2.4-12 hours, preferably 3.2-8 hours, more preferably 4 hours; the fourth base is sodium hydroxide, potassium hydroxide, or calcium hydroxide, preferably sodium hydroxide; the molar ratio of compound b-3 to 4-methoxybenzoyl chloride and the fourth base is 1:0.6-3:1.6-38.4, preferably... The ratio of compound C-3 to inorganic acid is selected as 1:0.8–2:2–9.6, more preferably 1:1.04:2.23; the solvent used in the esterification rearrangement reaction is tetrahydrofuran, methyltetrahydrofuran, or dimethyltetrahydrofuran, preferably tetrahydrofuran; the reaction temperature for the esterification rearrangement reaction is room temperature; the inorganic acid is concentrated sulfuric acid or concentrated hydrochloric acid, preferably concentrated sulfuric acid; the molar ratio of compound C-3 to inorganic acid is 1:0.24–4.32, preferably 1:0.28–1.44, 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 for the cyclization reaction is room temperature, and the reaction time is 1.8–12 h, preferably 2 h. The reaction time is 5-6 hours, more preferably 3 hours; the third Lewis acid is boron tribromide, boron trichloride, or aluminum trichloride, preferably boron tribromide; the molar ratio of 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 solution form, the solvent is dichloromethane, and the concentration of the third Lewis acid in the solution is 1.2-6 mol / L, preferably 1.6-4 mol / L, more preferably 2 mol / L; the third demethylation reaction is carried out at room temperature for 2.4-12 hours, preferably 3.2-8 hours, more preferably 4 hours.

[0111] Specifically, in synthetic route 3, the esterification rearrangement reaction is preferably carried out under inert gas protection; the third demethylation reaction is preferably carried out under inert gas protection; wherein, the inert gas is preferably nitrogen.

[0112] Furthermore, the present invention discloses a pharmaceutical composition comprising any one of the following: a compound represented by Formula I above, or a stereoisomer, geometric isomer, tautomer, nitride, hydrate, solvate, pharmaceutically acceptable salt, or prodrug, and one or more pharmaceutically acceptable carriers.

[0113] Specifically, the dosage form of the pharmaceutical composition is one or more of the following: subcutaneous injection, intradermal injection, spray, powder, external solution, lotion, liniment, ointment, plaster, paste, and patch.

[0114] Furthermore, this invention discloses a topical ointment containing the compound of Formula I or its stereoisomers, geometric isomers, tautomers, nitrides, hydrates, solvates, pharmaceutically acceptable salts, or prodrugs as active ingredients. It comprises an oil phase, an aqueous phase, and the active ingredient, with the following weight percentages for each material:

[0115] Oil phase:

[0116] Light liquid paraffin 0.6-3%, preferably 0.8-2%, more preferably 1.00%;

[0117] Cetyl alcohol 1.5-7.5%, preferably 2.0-5.0%, more preferably 2.54%;

[0118] Stearic acid 2.6-5.0%, preferably 3.2-4.4%, more preferably 3.80%;

[0119] Aqueous phase:

[0120] Glycerin 5.4-27%, preferably 7.2-18%, more preferably 8.92%;

[0121] Triethylamine 0.27-1.38%, preferably 0.35-0.92%, more preferably 0.46%;

[0122] Butylparaben: 0.06-3%, preferably 0.08-2%, more preferably 0.10%;

[0123] The active ingredient content is 0.6-12%, preferably 0.8-8%, and more preferably 1% to 4%.

[0124] The remainder is purified water;

[0125] The active ingredient is a compound represented by Formula I above, or a stereoisomer, geometric isomer, tautomer, nitride, hydrate, solvate, pharmaceutically acceptable salt, or prodrug thereof.

[0126] Furthermore, this invention discloses a method for preparing the above-mentioned topical ointment, comprising the following steps:

[0127] (1) Dissolve the active ingredient in a portion of purified water to obtain a drug solution; mix light liquid paraffin, cetyl alcohol, and stearic acid and heat to melt to obtain an oil phase; mix glycerol, triethylamine alcohol, butylparaben, and the remaining purified water and heat to melt to obtain an aqueous phase;

[0128] (2) Add the drug solution obtained in step (1) to the aqueous phase and mix well to obtain an aqueous phase containing the drug; add the aqueous phase containing the drug to the oil phase and stir until the temperature cools to room temperature to obtain the final product.

[0129] Specifically, the heating and melting process takes place at a temperature of 65–70°C.

[0130] The use of the compounds represented by Formula I above, or their stereoisomers, geometric isomers, tautomers, nitrides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs, or the pharmaceutical compositions described above, or the topical ointments described above, in the preparation of TRPV3 inhibitors are also within the scope of protection of this invention.

[0131] The use of the compound represented by Formula I above, or its stereoisomers, geometric isomers, tautomers, nitrides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs, or the pharmaceutical compositions described above, or the topical ointments described above, in the preparation of medicaments for treating TRPV3-mediated diseases is also within the scope of protection of this 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; more preferably, the TRPV3-mediated diseases are one or more of pruritus, alopecia, atopic dermatitis, psoriasis, and ulcerative colitis; even more preferably, the TRPV3-mediated diseases are psoriasis; the treatment of TRPV3-mediated psoriasis is to improve psoriatic lesions, psoriatic arthritis, or psoriatic pain.

[0132] The use of the compounds represented by Formula I above, or their stereoisomers, geometric isomers, tautomers, nitrides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs, or the pharmaceutical compositions described above, or the topical ointments described above, in the preparation of medications for treating pruritus, atopic dermatitis, and pain caused by kidney disease, is also within the scope of protection of this invention.

[0133] The use of the compounds represented by Formula I above, or their stereoisomers, geometric isomers, tautomers, nitrides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs, or the pharmaceutical compositions described above, or the topical ointments described above, in the preparation of drugs for treating kidney disease are also within the scope of protection of this invention.

[0134] The use of the compounds represented by Formula I above, or their stereoisomers, geometric isomers, tautomers, nitrides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs, or the pharmaceutical compositions described above, or the topical ointments described above, in the preparation of antifibrotic drugs are also within the scope of protection of this invention.

[0135] It should be noted that YHQ described in this invention is baicalein.

[0136] In the embodiments of the present invention, the purity of the intermediates prepared in each step is above 95%.

[0137] Unless otherwise stated, the term "alkyl" as used herein includes branched and straight-chain saturated aliphatic hydrocarbon groups having a specific number of carbon atoms, including all isomers. Common abbreviations for alkyl groups include methyl (e.g., "Me" or CH3), ethyl (e.g., "Et" or CH2CH3), propyl (e.g., "Pr" or CH2CH2CH3), butyl (e.g., "Bu" or CH2CH2CH2CH3," etc.). For example, "C..." 1-4 Alkyl (or "C1-C4 alkyl") refers to a straight-chain or branched alkyl group having a specific number of carbon atoms, including all isomers. 1-4 Alkyl groups include n-, iso-, secondary and tert-butyl, n- and isopropyl, ethyl and methyl. The term "C" is used in this context. 1-10 "Alkyl" and similar terms have similar meanings.

[0138] The term "alkoxy" refers to straight-chain and branched alkyl groups with a specified number of carbon atoms connected by oxygen bridges.

[0139] The term "halogen" (or "halogenated") refers to fluorine, chlorine, bromine, and iodine (or fluorinated (F), chlorinated (Cl), brominated (Br), and iodinated (I)).

[0140] The term "aryl" refers to aromatic monocyclic and polycyclic ring systems, in which the carbon rings are fused together or linked together by single bonds. Common aryl groups include phenyl, naphthyl, and biphenylene.

[0141] The term "heterocycle" refers to a ring structure composed of carbon atoms and non-carbon atoms. Examples of non-carbon atoms in the ring include nitrogen, oxygen, and sulfur. Common heterocyclic groups include pyridine, quinoline, tropane, phenothiazine, benzodiazepine, furan, pyrazolone, and pyrimidine.

[0142] The term "aromatic heterocycle" refers to a 5- or 6-membered monocyclic aromatic ring or a 7- to 12-membered bicyclic ring, which consists of a carbon atom and one or more heteroatoms selected from N, O, and S. Examples of aromatic heterocycles include pyridyl, pyrroloyl, pyrazinyl, pyrimidinyl, pyridazinyl, thiophene (or thiophenyl), thiazolyl, furanyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, azole, isozolyl, diazolyl, thiazolyl, isothiazolyl and thiadiazolyl, benzotriazolyl, indolyl, isoindolyl, indolyl, dihydroindolyl, isodihydroindolyl, quinoxalinyl, quinazolinyl, cenylyl, chromanyl, isochoryl, tetrahydroquinolinyl, quinolinyl, tetrahydroisoquinolinyl, isoquinolinyl, 2,3-dihydrobenzofuranyl, 2,3-dihydrobenzo-1,4-dienyl, imidazo(2,1-b)(1,3)thiazole and benzo-1,3-m-dioxanepentenyl.

[0143] The aryl group in the term "substituted aryl" is as defined above. When no substituent is specified for the substituted aryl group, the substituent may be selected from the following groups, including but not limited to: halogens, 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 ynyl, (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, heterocyclic alkyl, halogen-aryl, halogen-aralkyl, halogen-heterocyclic, halogen-heterocyclic alkyl, cyano-aryl, cyano-aralkyl, cyano-heterocyclic, and cyano-heterocyclic alkyl. The term "substituted phenyl" has a similar definition.

[0144] The term "pharmaceutically acceptable salt" refers to a salt prepared from a pharmaceutically acceptable, non-toxic alkali or acid. When the compounds of the present invention are acidic, their corresponding salts can be readily prepared from inorganic or organic bases. Salts derived from such inorganic bases include aluminum, ammonium, calcium, copper (copper and cuprous), iron, ferrous, lithium, magnesium, manganese (manganese and manganese), potassium, sodium, zinc, etc. Preferred are ammonium, calcium, magnesium, potassium, and sodium salts. Salts prepared from organic bases include primary, secondary, and tertiary amines derived from natural and synthetic sources. Pharmaceutically acceptable non-toxic organic 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, methylglucosamine, morpholine, piperazine, piperidine, polyamine resins, procaine, purines, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine, etc. When the compounds of the present invention are basic, their corresponding salts can be readily 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, hydroxyethylsulfonic acid, lactic acid, maleic acid, malic acid, mandelic acid, methanesulfonic acid, viscous acid, nitric acid, pyric acid, pantothenic acid, phosphoric acid, succinic acid, sulfuric acid, tartaric acid, p-toluenesulfonic acid, etc.

[0145] The term "solvate" refers to a variable stoichiometric complex 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 called a hydrate. Hydrates include, but are not limited to, hemihydrates, monohydrates, sesquihydrates, dihydrates, and trihydrates.

[0146] The present invention provides a pharmaceutical composition comprising a compound of Formula I or a stereoisomer, geometric isomer, tautomer, nitride, hydrate, solvate, pharmaceutically acceptable salt or prodrug, and one or more pharmaceutically acceptable carriers, diluents, and excipients.

[0147] In pharmaceutical compositions, the term "composition" includes products comprising an active ingredient and an inert component (pharmaceutical-acceptable excipient) constituting a carrier, as well as any product obtained directly or indirectly from a combination, complexation, or aggregation of two or more components, or from the decomposition of one or more components, or from other types of reactions or interactions of one or more components. Therefore, the pharmaceutical compositions of the present invention include any composition prepared by mixing a compound of formula I, other active ingredients, and pharmaceutically acceptable excipients.

[0148] The pharmaceutical compositions of the present invention comprise a compound of Formula I (or a pharmaceutically acceptable salt or solvate thereof) as an active ingredient, a pharmaceutically acceptable carrier, and optionally other therapeutic ingredients or adjuvants. The pharmaceutical compositions include those suitable for oral, rectal, topical, and parenteral (including subcutaneous, intramuscular, and intravenous) administration, although the most suitable route in any particular case depends on the specific subject, the nature and severity of the condition to which the active ingredient is administered. The pharmaceutical compositions can be prepared by any method known in the pharmaceutical field.

[0149] The active ingredient can be administered orally in solid or liquid dosage forms, such as capsules, tablets, lozenges, sugar lozenges, granules, and powders, and in liquid forms such as elixirs, syrups, emulsions, dispersions, and suspensions. The active ingredient can also be administered parenterally in sterile liquid dosage forms such as dispersions, suspensions, or solutions. Other dosage forms that can be used to administer the active ingredient include subcutaneous injections, intradermal injections, topical solutions, lotions, liniments, pastes, ointments, creams, drops, transdermal patches, or powders for topical administration; ophthalmic solutions or suspensions for ocular 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 the active ingredient and a powdered carrier, such as lactose, starch, cellulose derivatives, magnesium stearate, stearic acid, etc. Similar diluents can be used to prepare compressed tablets. Tablets and capsules can both be formulated as sustained-release products to provide sustained drug release over several hours. Compressed tablets can be sugar-coated or film-coated to mask any unpleasant taste and protect the tablet from air, or they can be enteric-coated for selective disintegration in the gastrointestinal tract. Liquid dosage forms for oral administration may contain colorants and flavoring agents to increase patient acceptability. Generally, water, suitable oils, saline, aqueous solutions of dextran (glucose), and related sugar solutions, and glycols such as propylene glycol or polyethylene glycol are suitable carriers for parenteral solutions. Solutions for parenteral administration preferably contain water-soluble salts of the active ingredient, suitable stabilizers, and buffers as needed. 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 may also be used. In addition, parenteral solutions may contain preservatives such as benzalkonium chloride, methylparaben, or propylparaben, and chlorobutanol. For inhalation administration, the compounds of the present invention can be conveniently delivered in aerosol form from pressurized packaging or nebulizers. The compounds can also be delivered in powder form, which can be inhaled with the aid of a powder inhaler device. A preferred delivery system for inhalation is a metered-dose inhaler (MDI) aerosol, which can be formulated as a suspension or solution of the compounds of Formula I or II in a suitable propellant, such as a fluorocarbon or hydrocarbon. For ophthalmic administration, ophthalmic formulations can be formulated as a solution or suspension of the compound of Formula I in a suitable ophthalmic carrier at a suitable weight percentage, thereby maintaining sufficient contact time between the compound and the ocular surface to allow the compound to penetrate into the cornea and internal regions of the eye.

[0150] 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 injections, and oral suspensions.

[0151] 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 combinations, 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, the second active ingredient including those known to be effective in increasing erythropoietin levels in patients.

[0152] Beneficial effects:

[0153] (1) This invention provides a new class of compounds that can be used as TRPV3 inhibitors. They 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.

[0154] (2) The compounds provided by the present invention have better solubility while having a comparable inhibition rate of calcium ion influx; at the same time, the compounds of the present invention have an inhibitory effect on the proliferation of human hepatic stellate cells LX-2 and have an anti-fibrotic effect. Detailed Implementation

[0155] The following non-limiting embodiments are intended to enable those skilled in the art to gain a more comprehensive understanding of the present invention, but do not limit the invention in any way. The following content is merely an exemplary description of the scope of protection claimed in this application. Those skilled in the art can make various changes and modifications to the invention based on the disclosed content, and such changes should also fall within the scope of protection claimed in this application.

[0156] The present invention will be further described below by way of specific embodiments. Unless otherwise specified, all chemical reagents used in the embodiments of the present invention are obtained through conventional commercial means.

[0157] Example 1: Preparation of 5,7,4'-trihydroxy-8-isopropylflavones

[0158]

[0159] Step 1: Preparation of 2-isopropyl-3,5-dimethoxyphenol

[0160] Substitution reaction: 3,5-Dimethoxyphenol (8 g, 51.89 mmol), chloroform (60 mL), 2-chloropropane (4.9 g, 62.26 mmol), and aluminum trichloride (18.7 g, 140.24 mmol) were added sequentially to a reaction flask. The mixture was stirred in an ice-water bath at 0 °C for 0.5 h, then slowly heated to room temperature for 4 h until the reaction was complete as monitored by TLC. After the reaction was complete, the reaction solution was cooled to room temperature, and 20 mL of purified water was slowly added dropwise to quench the reaction. The mixture was extracted with dichloromethane, washed 2 to 3 times with saturated brine, and the organic phase was separated. The organic phase was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the eluent was ethyl acetate:petroleum ether (v / v 1:40). Column chromatography yielded 4.0 g of a pale yellow solid, with a yield of 39%. Mass spectrometry data of the product 2-isopropyl-3,5-dimethoxyphenol: MS m / z (ESI) [M+H] + :197.11.

[0161] Step 2: Preparation of 2-hydroxy-3-isopropyl-4,6-dimethoxyacetophenone

[0162] Substitution reaction: 2-Isopropyl-3,5-dimethoxyphenol (2.4 g, 12.22 mmol), acetyl chloride (1.2 g, 14.66 mmol), chloroform (20 mL, redistilled), and aluminum trichloride (3.2 g, 24.46 mmol) prepared in step 1 were added sequentially to a reaction flask. The mixture was stirred at room temperature for 4 h until the reaction was complete as monitored by TLC. After the reaction was complete, 20 mL of purified water was slowly added dropwise in an ice-water bath to quench the reaction. The mixture was allowed to stand and separate into layers. The organic phase was washed 2 to 3 times with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The eluent was ethyl acetate:petroleum ether (v / v 1:30). Column chromatography yielded 1 g of a pale yellow oil, with a yield of 34%. Mass spectrometry data of the product 2-hydroxy-3-isopropyl-4,6-dimethoxyacetophenone: MS m / z (ESI) [M+H] + :239.28.

[0163] Step 3: Preparation of 1-(2-hydroxy-3-isopropyl-4,6-dimethoxyphenyl)-3-(4-methoxyphenyl)propane-1,3-dione

[0164] This step involves two reactions: the compound first undergoes esterification to generate 2-acetyl-6-isopropyl-3,5-dimethoxyphenyl-4-methoxybenzoate, and then the compound undergoes rearrangement.

[0165] Esterification 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 sequentially to a reaction flask, and the mixture was purged with nitrogen three times. The mixture was stirred at room temperature until the reaction was complete as monitored by TLC. After the reaction was complete, the pH was adjusted to neutral by adding dilute acetic acid, and the mixture was extracted with dichloromethane, washed 2 to 3 times with saturated brine, and the organic phase was separated. The organic phase was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the eluent was ethyl acetate:petroleum ether (volume ratio 1:8). Column chromatography yielded 1.2 g of a yellow solid, with a yield of 96%. Mass spectrometry 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.

[0166] Step 4: Preparation of 5,7,4'-trimethoxy-8-isopropylflavones

[0167] Ring-closure 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 sequentially to a reaction flask. The ring-closure reaction was carried out at room temperature with stirring for 3 h. LC-MS monitoring showed that the peak ratio of the starting material was less than or equal to 0.1%. Then, 15 mL of purified water was added, followed by 20 mL of dichloromethane solution. The mixture was stirred and allowed to stand for phase separation. The organic phase was washed 2 to 3 times with saturated brine, separated, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The eluent was ethyl acetate:petroleum ether (volume ratio 1:10) / methanol:dichloromethane (volume ratio 1:40). Column chromatography yielded 0.8 g of a yellow solid, with a yield of 84%. Mass spectrometry data of the product 5,7,4'-trimethoxy-8-isopropylflavone: MS m / z (ESI) [M+H] + :355.15.

[0168] Step 5: Preparation of 5,7,4'-trihydroxy-8-isopropylflavones

[0169] Demethylation reaction: 5,7,4'-trimethoxy-8-isopropylflavone (0.5 g, 1.41 mmol) prepared in step 4 and a 2 mol / L boron tribromide solution in dichloromethane (10 mL, containing 20 mmol BBr3) were added sequentially to a reaction flask. Under nitrogen protection, the demethylation reaction was carried out at room temperature with stirring for 4 h. LC-MS monitoring showed that the peak ratio of the starting material was less than or equal to 0.1%. The solvent was removed by rotary evaporation, and 4 mL of methanol was slowly added dropwise to quench the reaction. The methanol was evaporated to dryness, and 1.5 mL of methanol was added to dissolve the residue. Then, 3 mL of purified water was slowly added dropwise, and the mixture was stirred for 2 h. A large amount of orange-red solid precipitated. The mixture was filtered, and the filter cake was dried under vacuum 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 designated as compound 1, mp: 186.0-186.5 °C. The NMR and mass spectrometry data are shown below:

[0170] 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.

[0171] Example 2: Preparation of 2-(3-fluoro-4-hydroxyphenyl)-6,7-dihydroxyquinoline-4(1H)-one

[0172]

[0173] Step 1: Preparation of N-(2-acetyl-4,5-dimethoxyphenyl)-3-fluoro-4-methoxybenzamide

[0174] 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 sequentially to the reaction flask, and nitrogen gas was purged three times. The temperature was raised to 65 °C, and the acylation reaction was carried out for 4 h until the reaction of the starting materials was complete as monitored by TLC. After the reaction was complete, the reaction solution was cooled to room temperature, and 20 mL of purified water was slowly added dropwise to quench the reaction. The mixture was extracted with dichloromethane, washed 2-3 times with saturated brine, and the organic phase was separated. The solution was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the eluent was dichloromethane:ethyl acetate:petroleum ether (volume ratio 1:1:3). Column chromatography yielded 778 mg of a pale yellow solid, with a yield of 87.5%. Mass spectrometry 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.

[0175] Step 2: Preparation of 6,7-dimethoxy-2-(3-fluoro-4-methoxyphenyl)quinoline-4(1H)-one

[0176] Cyclization reaction: N-(2-acetyl-4,5-dimethoxyphenyl)-3-fluoro-4-methoxybenzamide (330 mg, 0.95 mmol, Mw = 347.342), 1,4-dioxane (15 mL), and sodium hydroxide (250 mg, 6.25 mmol, Mw = 40.00) prepared in step 1 were added sequentially to the reaction flask, and nitrogen gas was purged three times. The temperature was raised to 110 °C, and the intramolecular cyclization reaction was carried out for 4 h until the reaction of the starting materials was complete as monitored by TLC. After the reaction was completed, the reaction solution was cooled to room temperature, and the pH was adjusted to neutral by adding dilute acetic acid. The solution was extracted with dichloromethane, washed 2 to 3 times with saturated brine, and the organic phase was separated. The solution was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the eluent was dichloromethane:methanol (volume ratio of 100:7 to 100:10). Column chromatography yielded 252 mg of white solid, with a yield of 80.7%. Mass spectrometry data of the product 6,7-dimethoxy-2-(3-fluoro-4-methoxyphenyl)quinoline-4(1H)-one (Mw = 329.327): MS m / z (ESI) [M+H] + :330.31.

[0177] Step 3: Preparation of 6,7-dihydroxy-2-(3-fluoro-4-hydroxyphenyl)quinoline-4(1H)-one

[0178] Demethylation reaction: 210 mg of 6,7-dimethoxy-2-(3-fluoro-4-methoxyphenyl)quinoline-4(1H)-one (Mw = 329.327) prepared in step 2 and 2 mL of a 2 mol / L boron tribromide solution in dichloromethane (containing 34 mmol BBr) were added sequentially to a reaction flask. Under nitrogen protection, the demethylation reaction was carried out at room temperature with stirring for 4 h. LC-MS monitoring showed that the peak ratio of the starting material was less than or equal to 0.1%. The solvent was removed by rotary evaporation, and 4 mL of methanol was slowly added dropwise to quench the reaction. The methanol was evaporated to dryness, and 1.5 mL of methanol was added to dissolve the residue. Then, 3 mL of purified water was slowly added dropwise, and the mixture was stirred for 2 h. A large amount of orange-red solid precipitated. The mixture was 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 designated as compound 2, mp: 232.0-233.5℃. The NMR and mass spectrometry data are shown below:

[0179] 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.

[0180] Example 3: Preparation of 6,7-dihydroxy-2-(4-isopropylphenyl)quinoline-4(1H)-one

[0181]

[0182] The compound in this embodiment was prepared according to the preparation method of Example 2, with the material ratios remaining unchanged. The difference was that 3-fluoro-4-methoxybenzoyl chloride was replaced with 3.84 mmol of 4-isopropylbenzoyl chloride, yielding 128.9 mg of the product. The product is 6,7-dihydroxy-2-(4-isopropylphenyl)quinoline-4(1H)-one (C 18 H 17 NO3 (Mw = 295.338) is designated as compound 3, mp: 125.0-126.5℃. The NMR and mass spectrometry data are shown below:

[0183] 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 C-NMR (101MHz, DMSO) 166.50, 154.74, 152.66, 151.22, 148.21, 136.73, 129.79, 128.51, 127.46, 113.77, 104.57, 103.07, 102.01, 33.51, 23.71; MS m / z(ESI)[M+H] + :296.12.

[0184] Example 4: 2-(3,4-dichlorophenyl)-6,7-dihydroxyquinoline-4(1H)-one

[0185]

[0186] The preparation was carried out according to the method of Example 2, with the material ratios remaining unchanged, except that 3-fluoro-4-methoxybenzoyl chloride was replaced with 3.84 mmol of 3,4-dichlorobenzoyl chloride, yielding 120.3 mg of product. The product 2-(3,4-dichlorophenyl)-6,7-dihydroxyquinoline-4(1H)-one (C 15 H9C l2 NO3 (Mw = 322.141) is designated as compound 4, mp: 206.2-207.5℃. The NMR and mass spectrometry data are shown below:

[0187] 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.

[0188] Example 5: Preparation of N-(2-acetyl-4,5-dihydroxyphenyl)-3-fluoro-4-hydroxybenzamide

[0189]

[0190] The product was prepared according to steps 1 (acylation) and 3 (demethylation) in Example 2, with the material ratios remaining constant. Acylation was performed using 3.84 mmol of 3-fluoro-4-methoxybenzoyl chloride with 2-amino-4,5-dimethoxyacetophenone, followed by demethylation, yielding 201.6 mg of the product. The product is N-(2-acetyl-4,5-dihydroxyphenyl)-3-fluoro-4-hydroxybenzamide (C... 15 H 12 FNO5 (Mw = 305.261) is designated as compound 5, mp: 292.0-293.5℃. NMR and mass spectrometry data are as follows:

[0191] 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.

[0192] Example 6: N-(2-acetyl-4,5-dihydroxyphenyl)-3,4-dichlorobenzamide

[0193]

[0194] The preparation was carried out according to steps 1 (acylation reaction) and 3 (demethylation reaction) in Example 2, with the material ratios remaining constant. The difference was that 3-fluoro-4-methoxybenzoyl chloride was replaced with 3.84 mmol of 3,4-dichlorobenzoyl chloride for acylation with 2-amino-4,5-dimethoxyacetophenone, followed by demethylation, yielding 191.6 mg of the product. The product N-(2-acetyl-4,5-dihydroxyphenyl)-3,4-dichlorobenzamide (C...) 15 H 11 C l2 NO4 (Mw = 340.156), denoted as compound 6, has the following NMR and mass spectrometry data:

[0195] 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.

[0196] Example 7: Water solubility test

[0197] 1. Reagents and consumables

[0198] Table 1 Reagents and Consumables

[0199]

[0200]

[0201] 2. Experimental Methods

[0202] Prepare 10 mmol / L stock solutions of analytes 1-6 (prepared in Examples 1-6), YHQ (baicalein, self-made), and the reference standard progesterone using DMSO. Add 30 μL of the 10 mmol / L stock solution to each well of a 96-well plate, followed by 970 μL of phosphate buffer (pH 7.4). Stir at 25°C and 1100 rpm for 2 hours. Filter the solution, transfer 5 μL of the filtrate to a new sample plate, and add 5 μL of DMSO and 490 μL of 50% ACN (acetonitrile) solution. Prepare the standard solution with phosphate buffer (pH 7.4) to achieve a final progesterone concentration of 3 μmol / L. Calculate the saturated solubility of the analytes using qualitative and quantitative analysis of the standard peaks at known concentrations via LC-MS, as shown in Table 2.

[0203] Table 2 Solubility of each compound

[0204]

[0205] Example 8: Biological Test

[0206] 1. Calcium ion concentration detection

[0207] 1.1 Reagents and Consumables

[0208] Table 3 Reagents and Consumables

[0209]

[0210]

[0211] 1.2 Test Methods

[0212] (1) Preparation of compound stock solution

[0213] Preparation of stock solutions for compounds 1-6: Each compound powder (prepared in Examples 1-6) was accurately weighed and prepared into a 1 mM stock solution with DMSO. After being dispensed, the stock solutions were stored at -80°C and had a shelf life of 1 year.

[0214] Preparation of TRPV3 agonist 2-APB stock solution: Weigh 1.3505 mg 2-APB powder, add 200 μL DMSO solution to prepare a 30 mM stock solution, aliquot and store at -80℃.

[0215] Preparation of stock solution of TRPV3 inhibitor YHQ (baicalein, self-made): Weigh 1.7174 mg of baicalein powder, add 200 μL of DMSO solution to prepare a 1 mM stock solution, aliquot and store at -80℃.

[0216] (2) Buffer preparation

[0217] Preparation of extracellular buffer solution for TRPV3 calcium signal detection (abbreviated as: extracellular buffer solution): Mix the materials in Table 4 and adjust the pH to 7.4 with NaOH.

[0218] Table 4. Dosage of each component in the external solution for 1000mL TRPV3 calcium signal detection

[0219]

[0220] Preparation of calcium fluorescent dye: Add 228 μL of DMSO to 1 mg of Fluo-4 powder to prepare a 4 mM stock solution. Wrap in aluminum foil to protect from light and vortex. Dispense into 200 μL centrifuge tubes, 10 μL per tube.

[0221] Preparation of PBS buffer: Mix the components in Table 5, and then adjust the pH to 7.2-7.4 using HCl and NaOH.

[0222] Table 5. Components and dosage in 1000 mL PBS buffer

[0223]

[0224] (3) Cell Culture

[0225] HEK-293 cells were cultured in DMEM complete medium containing 10% FBS, 1% P / S, and 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 medium were preheated and filtered in a biosafety cabinet. The medium in the flask was discarded. The remaining medium was washed twice with PBS and discarded. Trypsin-EDTA digestion solution was added to digest the cells until they became rounded and the cytoplasm retracted. The trypsin-EDTA digestion solution was then discarded. DMEM complete medium was added to stop the digestion, and the cells at the bottom of the flask were dispersed. The cell suspension was transferred to a small dish (density 50%-60%), DMEM complete medium was added, and the mixture was slowly mixed. The dish was incubated for 6 hours before being used for cell transfection.

[0226] (4) Cell transfection

[0227] Filter the preheated PBS buffer (pH = 7.2-7.4) and serum-depleted medium (opti-MEM) in a biosafety cabinet. Discard the original medium in the dish (the final result obtained in "(3) Cell Culture"), wash twice with PBS, add opti-MEM, and incubate in a cell culture incubator for 1 hour. Take the h-TRPV3 plasmid and Lipofectamine. TM Prepare the transfection system according to the instructions for use with the 2000 transfection reagent. After 1 hour, add the solution of plasmid and transfection reagent dropwise to the small dish. Incubate in a cell culture incubator for 6-8 hours, then replace with normal DMEM complete medium for subsequent plate-laying experiments.

[0228] (5) Cell plating

[0229] Pre-coating of 96-well plates with poly-L-lysine: A 10 mg / mL poly-L-lysine stock solution was prepared using sterile water and dispensed into 0.2 mL centrifuge tubes (10 μL per tube), which were then stored in an ultra-low temperature freezer. Before use, the poly-L-lysine stock solution was removed from the freezer and diluted with sterile water to a final concentration of 10 μg / mL. The diluted poly-L-lysine solution was filtered, and 60 μL / well was coated onto each well of a 96-well black-walled permeable plate. The plate was incubated at 37°C with 5% CO2 for 1 hour. Before cell seeding, the poly-L-lysine solution in the wells was discarded.

[0230] 14 hours before the real-time fluorescence experiment, TRPV3-HEK 293 transiently transfected cells were washed with PBS, digested with trypsin-EDTA digestion solution, and then transferred to 96-well black-walled permeabilized plates coated with poly-L-lysine at a density of 100 μL / well (2 × 10⁻⁶). 4 Cells / pores.

[0231] (6) Add 60 μL of dye (the “calcium fluorescent dye” prepared under the “(2) buffer preparation” section), put the cell plate back into the incubator, and load the dye for 60 min.

[0232] (7) Remove the cell plate and add 140 μL of extracellular buffer (prepared under "(2) Buffer Preparation") to each well, making the volume 200 μL. Wash with 150 μL of extracellular buffer, repeating 3-4 times. After the last wash, aspirate the 150 μL solution and add 100 μL of extracellular buffer to make the volume 150 μL of extracellular fluid in each well. After washing, place the cell plate in the Read position of the FLIPR and stabilize for 5 min.

[0233] The test compounds (a mixture of inhibitor YHQ and compounds 1-6 with extracellular fluid to a final concentration of 1.0 μmol / L) and the agonist (a mixture of 2-APB and extracellular fluid to a final concentration of 30 μmol / L) were added at 60 seconds and 360 seconds after the start of signal acquisition, respectively, with a signal acquisition interval of 1 second. Fluorescence values ​​at different time points were recorded at a wavelength of 535 nm. Fluorescence values ​​were obtained from Screenwork 4.0 software (Molecular device, CA, USA), the data were exported and processed to obtain the inhibition rate of the test compounds. The results are shown in Table 6 below.

[0234] Table 6. Inhibition rate of calcium ions by each compound

[0235] Compound numbering Inhibition rate % (concentration 1.0 μM) Inhibitor YHQ (baicalin) (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)％

[0236] As shown in the table above, compounds 1-3 prepared in this invention can inhibit calcium ion influx at a single concentration of 1.0 μM, exhibiting significant TRPV3 inhibitory activity.

[0237] Example 9: Preparation of a topical ointment

[0238] Topical ointments consist of an oil phase, an aqueous phase, and active ingredients, with the following weight percentages for each component:

[0239] Oil phase: 1.00% light liquid paraffin; 2.54% cetyl alcohol; 3.80% stearic acid;

[0240] Aqueous phase: glycerol 8.92%; triethylamine 0.46%; butylparaben 0.10%;

[0241] The remainder is purified water;

[0242] Active ingredients: Compounds 1 to 6 (prepared in Examples 1 to 6, respectively).

[0243] Preparation of topical ointments:

[0244] (1) Dissolve compounds 1 to 6 in a portion of purified water to obtain a drug solution; mix 1.00% light liquid paraffin, 2.54% cetyl alcohol, and 3.80% stearic acid and heat to melt at 65-70°C to obtain an oil phase; mix 8.92% glycerol, 0.46% triethylamine alcohol, 0.10% butylparaben, and the remaining purified water and heat to melt at 65-70°C to obtain an aqueous phase.

[0245] (2) Add the drug solution obtained in step (1) to the aqueous phase and mix well to obtain an aqueous phase containing the drug; add the aqueous phase containing the drug to the oil phase and stir quickly until the temperature is cooled to room temperature to obtain pale yellow topical ointments containing 1% of compounds 1 to 6.

[0246] Example 10: Pharmacological Evaluation of the Compound

[0247] 1. Experimental Objective

[0248] The purpose of this experiment was to evaluate the effect of topical administration of compounds 1-6 (prepared from Examples 1-6, respectively) on the efficacy of imiquimod in inducing psoriasis in C57BL / 6 mice.

[0249] 2. Laboratory animals

[0250] Twenty-five SPF-grade C57BL / 6 mice, 11 weeks old, male, and weighing 25–30 g, were purchased from the Comparative Medicine Center of Yangzhou University [Experimental Animal Production License No.: SCXK(Su)2022-0009]. They were housed at room temperature (23±2)℃, humidity 55%, with 12 hours of light and 12 hours of darkness, and free access to food and water for 7 days to allow for acclimatization. The experiment was approved by the Institute of Hygiene and Environmental Technology, Soochow University (Experimental Animal Use License No.: SYXK(Su)2022-0057).

[0251] 3. Information on the test compound and positive control drug

[0252] Imiquimod ointment (Imiquimod, IMQ; batch number: 40220601, Sichuan Mingxin Pharmaceutical Co., Ltd., 50 mg / g); Mometasone furoate ointment (Momet; batch number: 221008, Shanghai Minhang Pharmaceutical Co., Ltd., white semi-solid ointment, 3 g, 1 mg / g); 1% Compound 2 ointment (prepared in Example 9); Baicalein (YHQ, self-made).

[0253] 4. Animal grouping and animal model establishment

[0254] Twenty-five male SPF-grade C57BL / 6 mice were shaved, with a back hair area of ​​approximately 2×3 cm. Residual hair was removed with depilatory cream. After 24 hours, the mice were randomly divided into five groups: control group, 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 five mice in each group.

[0255] Except for the blank control group, the other groups applied 62.5 mg of IMQ to the skin on the back of mice to establish the model every morning; 8 hours after administration, the corresponding drug ointment with the same dose (100 mg for each cream and 100 mg for YHQ) was administered for 5 consecutive days.

[0256] Preparation of blank matrix: The composition ratio and preparation method of the blank matrix are the same as in Example 9, except that no active ingredient compound is added.

[0257] 5. Project Testing

[0258] (1) On days 1, 3, and 5, at the same time on the first day of the experiment, the severity of the mouse skin lesions was assessed using the PASI scoring scale, focusing on three aspects: erythema, scaling, and skin thickening. PASI scoring scale: 0-4 points, with severity levels ranging from none to mild, moderate, and severe. The total score was the sum of the three indicators (0-12 points).

[0259] (2) Skin tissue from the drug-treated area on the back of mice was photographed on days 1, 3, and 5. After the modeling and drug administration, skin tissue from the drug-treated area on the back of mice was collected, washed with physiological saline, fixed in 4% (w / v) paraformaldehyde solution for 48 hours, and then embedded in paraffin. The tissue was then cut into 3 μm paraffin sections, stained with hematoxylin and eosin as usual, and the pathological changes in the skin were observed and photographed under a microscope. Epidermal thickness was calculated using the Image-Pro plus pathological image analysis system.

[0260] 6. Results and Analysis

[0261] Animal experiments showed that, under the same dosage conditions, compound 2 could reduce the PASI score of imiquimod-induced psoriasis in mice, improve the clinicopathological appearance of psoriasis mice, and had a good inhibitory effect on abnormal epidermal thickening in psoriasis mice. The pharmacodynamic effect on psoriasis mice was: compound 2 > baicalein (YHQ), as shown in Table 7.

[0262] Table 7. Efficacy evaluation of compound 2 in mice with psoriasis.

[0263]

[0264] Example 11: Inhibitory effect of compound 2 on the proliferation of human hepatic stellate cells LX-2

[0265] 1. Method

[0266] 1.1 LX-2 cell culture

[0267] LX-2 cells (provided by Beijing Beina Chuanglian Biotechnology Research Institute) were cultured in culture flasks provided by Wuhan Pronosai Life Science Technology Co., Ltd., containing RPMI 1640 medium. The medium contained 10% fetal bovine serum (Zhejiang Tianhang Biotechnology Co., Ltd.), 100 mg / mL streptomycin, and 100 kU / L penicillin. The flasks were incubated in a 5% CO2 incubator (Binder GmbH, Germany) at saturated humidity and 37°C.

[0268] 1.2 MTT assay for cell proliferation activity

[0269] LX-2 cells in logarithmic growth phase were harvested, digested with 0.25% trypsin, and then the cell density was adjusted to 5 × 10⁶ cells / year using RPMI 1640 medium (10% FBS + 1% penicillin and antibiotic). 4 Cells were inoculated at a density of 100 μL / mL into 96-well plates. Cells were divided into four groups: a normal control group (no drug), a TGF-β1 group (TGF-β1 added to the plate at a concentration of 10 ng / mL), a baicalein (YHQ) group (YHQ added to the plate at a concentration of 10 μM), and a compound 2 group (compound 2 prepared in Example 2 added to the plate at a concentration of 10 μM). Each group had six replicates, with 100 μL of cell suspension in each well. A blank zeroing well was also included. 200 μL of sterile PBS buffer was added around each cell well. Cells were cultured at 37°C and 5% CO2 saturated humidity for 24 h. Then, 20 μL of MTT was added to each well, and the cells were cultured under the same conditions for 3 h. The culture medium was then aspirated, and 100 μL of DMSO was added and shaken for 10 min. The absorbance (A) at 490 nm was measured using a microplate reader to calculate the cell proliferation rate. Cell proliferation rate = (A experimental group - A zero-adjustment group) / (A control group - A zero-adjustment group) × 100%.

[0270] The experimental results are shown in Table 8. As can be seen from Table 8, compound 2 prepared in Example 2 has a good inhibitory effect on the proliferation of LX-2 cells.

[0271] Table 8. Inhibition data of different drugs on LX-2 cells

[0272] sample normal group TGF-β1 group Example 2 Compound 2 YHQ Group Cell viability / % 100 120 50 75

[0273] 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 principles of the present invention should be included within the protection scope of the present invention.

[0274] This invention provides a transient receptor potential cation channel TRPV3 inhibitor and its application, along with a conceptual approach and method. Many methods and approaches exist for implementing this technical solution; the above description is merely a preferred embodiment of the invention. It should be noted that those skilled in the art can make various improvements and modifications without departing from the principles of this invention, and these improvements and modifications should also be considered within the scope of protection of this invention. All components not explicitly stated in this embodiment can be implemented using existing technologies.

Claims

1. A compound as shown in Formula 2 or a pharmaceutically acceptable salt thereof; 2. The compound of claim 1, wherein, It was prepared by the following method, and the following synthetic route was selected:

3. A process for the preparation of a compound according to claim 1, characterized in that, Choose the following synthesis route:

4. A pharmaceutical composition, characterized by, It comprises any one of the compounds of claim 1 or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers.

5. The pharmaceutical composition of claim 4, wherein, The dosage form of the pharmaceutical composition is one or more of the following: subcutaneous injection, intradermal injection, spray, powder, external solution, lotion, liniment, ointment, plaster, paste, and patch.

6. A topical ointment containing the compound of claim 1 or a pharmaceutically acceptable salt thereof as the active ingredient, characterized in that, Including oil phase, aqueous phase, and active ingredients, the weight percentage of each material is as follows: Oil phase: Light liquid paraffin 0.6-3%; Cetyl alcohol 1.5-7.5%; Stearic acid 2.6-5.0%; Aqueous phase: Glycerin 5.4-27%; Triethylamine 0.27-1.38%; Butylparaben 0.06-3%; Active ingredient 0.6-12%; The remainder is purified water; The active ingredient is the compound of claim 1 or a pharmaceutically acceptable salt thereof.

7. The topical ointment according to claim 6, characterized in that, Including oil phase, aqueous phase, and active ingredients, the weight percentage of each material is as follows: Oil phase: Light liquid paraffin 0.8-2%; Cetyl alcohol 2.0-5.0%; Stearic acid 3.2-4.4%; Aqueous phase: Glycerin 7.2-18%; Triethylamine 0.35-0.92%; Butylparaben 0.08-2%; Active ingredient 0.8-8%; The remainder is purified water; The active ingredient is the compound of claim 1 or a pharmaceutically acceptable salt thereof.

8. The topical ointment according to claim 6, characterized in that, Including oil phase, aqueous phase, and active ingredients, the weight percentage of each material is as follows: Oil phase: Light liquid paraffin 1.00%; Cetyl alcohol 2.54%; Stearic acid 3.80%; Aqueous phase: Glycerin 8.92%; Triethylamine 0.46%; Butylparaben 0.10%; Active ingredient 1%–4%; The remainder is purified water; The active ingredient is the compound of claim 1 or a pharmaceutically acceptable salt thereof.

9. The topical ointment according to any one of claims 6 to 8, characterized in that, The preparation method of the topical ointment includes the following steps: (1) Dissolve the active ingredient in a portion of purified water to obtain a drug solution; mix light liquid paraffin, cetyl alcohol, and stearic acid and heat to melt to obtain an oil phase; mix glycerol, triethylamine alcohol, butylparaben, and the remaining purified water and heat to melt to obtain an aqueous phase; (2) Add the drug solution obtained in step (1) to the aqueous phase and mix well to obtain an aqueous phase containing the drug; add the aqueous phase containing the drug to the oil phase and stir until the temperature cools to room temperature to obtain the final product.

10. The use of the compound of claim 1 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 4, or the topical ointment of claim 6 in the preparation of a TRPV3 inhibitor, or in the preparation of a medicament for treating pruritus, atopic dermatitis, and pain caused by kidney disease, or in the preparation of a medicament for treating kidney disease, or in the preparation of a medicament for treating antifibrotic diseases.

11. Use of the compound of claim 1 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 4, or the topical ointment of claim 6 in the preparation of a medicament for treating TRPV3-mediated diseases.

12. The use according to claim 11, characterized in that, The treatment of TRPV3-mediated diseases involves inhibiting the increased calcium ion influx caused by TRPV3 activation.

13. The use according to claim 11 or 12, characterized in that, The TRPV3-mediated diseases are one or more of the following: pruritus, hair loss, atopic dermatitis, psoriasis, and ulcerative colitis.

14. The use according to claim 11 or 12, characterized in that, The TRPV3-mediated disease is psoriasis.

15. The use according to claim 14, characterized in that, The treatment for TRPV3-mediated psoriasis aims to improve psoriasis-like lesions, psoriatic arthritis, or psoriasis pain.

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