TRPA1 regulator as well as pharmaceutical composition and application thereof

Abstract

The invention belongs to the technical field of biological medicines, and particularly relates to a TRPA1 regulator as well as a pharmaceutical composition and application thereof. The invention provides a TRPA1 regulator, the structural general formula of which is as shown in formula I, # imgabs0. The TRPA1 regulator provided by the invention shows good biological activity and good druggability in animal bodies, can achieve the purpose of inhibiting TRPA1 current through desensitization, and inhibits the generation of neuron action potential, thereby treating nervous system diseases related to the TRPA1 regulator.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine technology, and specifically relates to a TRPA1 regulator and a pharmaceutical composition and application thereof. Background Art

[0002] It has been reported that TRPA1 modulators such as AITC have obvious irritation and poor drugability; GEN551 cannot desensitize TRPA1 channels, and long-term activation of TRPA1 channels produces pain responses in animals. Therefore, it is necessary to provide an improved TRPA1 modulator to solve the above problems. Summary of the invention

[0003] The purpose of the present invention is to provide a TRPA1 modulator and a pharmaceutical composition and application thereof. The compound can inhibit TRPA1 current through desensitization, inhibit the generation of neuronal action potentials, and thus treat related nervous system diseases.

[0004] To achieve the above objectives, in a first aspect, the present invention provides a TRPA1 modulator, the general structural formula of which is shown in Formula I:

[0005]

[0006] wherein Ring A is selected from C6-12 aryl or 5-10 membered heteroaryl, wherein the aryl or heteroaryl is optionally substituted with 1-3 halogen, amino, hydroxy, carboxyl, nitro, trifluoromethyl, amide, methoxy, cyano or C1-4 alkyl;

[0007] Wherein, R1 is selected from hydrogen, halogen, cyano, nitro, amino, hydroxyl, carboxyl, trifluoromethyl, amide, methoxy, C1-C4 alkyl, C1-C4 alkoxy, C3-C6 cycloalkyl;

[0008] wherein R2 and R3 are selected from H or C1-C6 alkyl, or C1-C6 alkyl, C3-C6 cycloalkyl, C1-C6 alkylcarbonyl, C1-C6 alkoxycarbonyl, C1-C6 alkylaminocarbonyl or C2-C8 alkenyl which may be optionally substituted with hydroxyl, amino, C1-C4 alkoxy, C1-C4 alkylcarbonyl, halogen atom, C6-10 aryl or halogenated C6-10 aryl; -C2-C8 alkenyl, C5-C7 cycloalkenyl or C2-C8 alkynyl substituted by a C4 alkylcarbonyl, a C1-C4 alkoxy, a halogen atom, a C6-C10 aryl or a halogenated C6-C10 aryl group; C2-C8 alkynyl or C6-C10 aryl which may be optionally substituted by a hydroxyl, an amino, a C1-C4 alkylcarbonyl, a C1-C4 alkoxy, a halogen atom, a C6-C10 aryl or a halogenated C6-C10 aryl group.

[0009] Furthermore, the TRPA1 modulator has a general structural formula as shown in Formula II:

[0010]

[0011] Wherein, R1 is selected from hydrogen, halogen, cyano, nitro, amino, hydroxyl, carboxyl, trifluoromethyl, amide, methoxy, C1-C4 alkyl, C1-C4 alkoxy, C3-C6 cycloalkyl;

[0012] wherein R2 and R3 are selected from H or C1-C6 alkyl or C1-C6 alkyl which may be optionally substituted by hydroxyl, amino, C1-C4 alkoxy, C1-C4 alkylcarbonyl, halogen atom, C6-10 aryl or halogenated C6-10 aryl, C3-C6 cycloalkyl, C1-C6 alkylcarbonyl, C1-C6 alkoxycarbonyl, C1-C6 alkylaminocarbonyl, C2-C8 alkenyl, which may be optionally substituted by hydroxyl, amino, C1-C4 alkoxy, C1-C4 alkylcarbonyl, halogen atom, C6-10 aryl or halogenated C6-10 aryl - C2-C8 alkenyl substituted by a C4-alkylcarbonyl group, a C1-C4-alkoxy group, a halogen atom, a C6-C10 aryl group or a halogenated C6-C10 aryl group, a C5-C7 cycloalkenyl group, a C2-C8 alkynyl group, a C2-C8 alkynyl group or a C6-C10 aryl group which may be optionally substituted by a hydroxyl group, an amino group, a C1-C4-alkylcarbonyl group, a C1-C4-alkoxy group, a halogen atom, a C6-C10 aryl group or a halogenated C6-C10 aryl group;

[0013] Among them, X, Y = C, O, N, S.

[0014] Furthermore, the TRPA1 modulator has a general structural formula as shown in Formula III:

[0015]

[0016] Wherein, R1 is selected from hydrogen, halogen, cyano, nitro, amino, hydroxyl, carboxyl, trifluoromethyl, amide, methoxy, C1-C4 alkyl, C1-C4 alkoxy, C3-C6 cycloalkyl;

[0017] wherein R2 and R3 are selected from H, C1-C6 alkyl, C1-C6 alkyl which may be optionally substituted by hydroxyl, amino, C1-C4 alkoxy, C1-C4 alkylcarbonyl, halogen atom, C6-10 aryl or halogenated C6-10 aryl, C3-C6 cycloalkyl, C1-C6 alkylcarbonyl, C1-C6 alkoxycarbonyl, C1-C6 alkylaminocarbonyl, C2-C8 alkenyl, which may be optionally substituted by hydroxyl, amino, C1-C4 alkoxy, C1-C4 alkylcarbonyl, halogen atom, C6-10 aryl or halogenated C6-10 aryl - C2-C8 alkenyl substituted by a C4-alkylcarbonyl group, a C1-C4-alkoxy group, a halogen atom, a C6-C10 aryl group or a halogenated C6-C10 aryl group, a C5-C7 cycloalkenyl group, a C2-C8 alkynyl group, a C2-C8 alkynyl group or a C6-C10 aryl group which may be optionally substituted by a hydroxyl group, an amino group, a C1-C4-alkylcarbonyl group, a C1-C4-alkoxy group, a halogen atom, a C6-C10 aryl group or a halogenated C6-C10 aryl group;

[0018] Furthermore, the TRPA1 modulator has a general structural formula as shown in Formula IV:

[0019]

[0020] Wherein, R1 is selected from hydrogen, halogen, cyano, nitro, amino, hydroxyl, carboxyl, trifluoromethyl, amide, methoxy, C1-C4 alkyl, C1-C4 alkoxy, C3-C6 cycloalkyl;

[0021] wherein R2 is selected from H, C1-C6 alkyl, C1-C6 alkyl which may be optionally substituted by hydroxyl, amino, C1-C4 alkoxy, C1-C4 alkylcarbonyl, halogen atom, C6-10 aryl or halogenated C6-10 aryl, C3-C6 cycloalkyl, C1-C6 alkylcarbonyl, C1-C6 alkoxycarbonyl, C1-C6 alkylaminocarbonyl, C2-C8 alkenyl, which may be optionally substituted by hydroxyl, amino, C1-C4 alkoxy, C1-C4 alkylcarbonyl, halogen atom, C6-10 aryl or halogenated C6-10 aryl C2-C8 alkenyl substituted by an alkylcarbonyl group, a C1-C4 alkoxy group, a halogen atom, a C6-C10 aryl group or a halogenated C6-C10 aryl group, a C5-C7 cycloalkenyl group, a C2-C8 alkynyl group, a C2-C8 alkynyl group or a C6-C10 aryl group which may be optionally substituted by a hydroxyl group, an amino group, a C1-C4 alkylcarbonyl group, a C1-C4 alkoxy group, a halogen atom, a C6-C10 aryl group or a halogenated C6-C10 aryl group;

[0022] Furthermore, the TRPA1 modulator has a general structural formula as shown in Formula V:

[0023]

[0024] Wherein, R1 is selected from hydrogen, halogen, cyano, nitro, amino, hydroxyl, carboxyl, trifluoromethyl, amide, methoxy, C1-C4 alkyl, C1-C4 alkoxy, C3-C6 cycloalkyl;

[0025] wherein R2 is selected from H, C1-C6 alkyl, C1-C6 alkyl which may be optionally substituted by hydroxyl, amino, C1-C4 alkoxy, C1-C4 alkylcarbonyl, halogen atom, C6-10 aryl or halogenated C6-10 aryl, C3-C6 cycloalkyl, C1-C6 alkylcarbonyl, C1-C6 alkoxycarbonyl, C1-C6 alkylaminocarbonyl, C2-C8 alkenyl, which may be optionally substituted by hydroxyl, amino, C1-C4 alkoxy, C1-C4 alkylcarbonyl, halogen atom, C6-10 aryl or halogenated C6-10 aryl C2-C8 alkenyl substituted by an alkylcarbonyl group, a C1-C4 alkoxy group, a halogen atom, a C6-C10 aryl group or a halogenated C6-C10 aryl group, a C5-C7 cycloalkenyl group, a C2-C8 alkynyl group, a C2-C8 alkynyl group or a C6-C10 aryl group which may be optionally substituted by a hydroxyl group, an amino group, a C1-C4 alkylcarbonyl group, a C1-C4 alkoxy group, a halogen atom, a C6-C10 aryl group or a halogenated C6-C10 aryl group;

[0026] Furthermore, the TRPA1 modulator has a general structural formula as shown in Formula VI:

[0027]

[0028] Wherein, R1 is selected from hydrogen, halogen, cyano, nitro, amino, hydroxyl, carboxyl, trifluoromethyl, amide, methoxy, C1-C4 alkyl, C1-C4 alkoxy, C3-C6 cycloalkyl;

[0029] wherein R2 is selected from H, C1-C6 alkyl, C1-C6 alkyl which may be optionally substituted by hydroxyl, amino, C1-C4 alkoxy, C1-C4 alkylcarbonyl, halogen atom, C6-10 aryl or halogenated C6-10 aryl, C3-C6 cycloalkyl, C1-C6 alkylcarbonyl, C1-C6 alkoxycarbonyl, C1-C6 alkylaminocarbonyl, C2-C8 alkenyl, which may be optionally substituted by hydroxyl, amino, C1-C4 alkoxy, C1-C4 alkylcarbonyl, halogen atom, C6-10 aryl or halogenated C6-10 aryl C2-C8 alkenyl substituted by an alkylcarbonyl group, a C1-C4 alkoxy group, a halogen atom, a C6-C10 aryl group or a halogenated C6-C10 aryl group, a C5-C7 cycloalkenyl group, a C2-C8 alkynyl group, a C2-C8 alkynyl group or a C6-C10 aryl group which may be optionally substituted by a hydroxyl group, an amino group, a C1-C4 alkylcarbonyl group, a C1-C4 alkoxy group, a halogen atom, a C6-C10 aryl group or a halogenated C6-C10 aryl group;

[0030] Furthermore, the TRPA1 modulator has a general structural formula as shown in Formula VII:

[0031]

[0032] Wherein, R1 is selected from hydrogen, halogen, cyano, nitro, amino, hydroxyl, carboxyl, trifluoromethyl, amide, methoxy, C1-C4 alkyl, C1-C4 alkoxy, C3-C6 cycloalkyl;

[0033] wherein R2 is selected from H, C1-C6 alkyl, C1-C6 alkyl which may be optionally substituted by hydroxyl, amino, C1-C4 alkoxy, C1-C4 alkylcarbonyl, halogen atom, C6-10 aryl or halogenated C6-10 aryl, C3-C6 cycloalkyl, C1-C6 alkylcarbonyl, C1-C6 alkoxycarbonyl, C1-C6 alkylaminocarbonyl, C2-C8 alkenyl, which may be optionally substituted by hydroxyl, amino, C1-C4 alkoxy, C1-C4 alkylcarbonyl, halogen atom, C6-10 aryl or halogenated C6-10 aryl C2-C8 alkenyl substituted by an alkylcarbonyl group, a C1-C4 alkoxy group, a halogen atom, a C6-C10 aryl group or a halogenated C6-C10 aryl group, a C5-C7 cycloalkenyl group, a C2-C8 alkynyl group, a C2-C8 alkynyl group or a C6-C10 aryl group which may be optionally substituted by a hydroxyl group, an amino group, a C1-C4 alkylcarbonyl group, a C1-C4 alkoxy group, a halogen atom, a C6-C10 aryl group or a halogenated C6-C10 aryl group;

[0034] Furthermore, the TRPA1 modulator is selected from the following compounds:

[0035]

[0036]

[0037] Based on the same inventive concept, the present invention also provides a pharmaceutical composition, the active ingredients of which include the TRPA1 modulator represented by the above formula (I) and its stereoisomers, prodrugs, solvates, hydrates and pharmaceutically acceptable inorganic acids or organic acid salts thereof;

[0038] Wherein, the inorganic acid includes but is not limited to any of hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfonic acid, carbonic acid, sulfuric acid, phosphoric acid, nitric acid and perchloric acid;

[0039] Wherein, the organic acid includes but is not limited to any of maleic acid, succinic acid, citric acid, tartaric acid, fumaric acid, formic acid, acetic acid, propionic acid, malonic acid, oxalic acid, benzoic acid, phthalic acid, methanesulfonic acid, benzenesulfonic acid, toluenesulfonic acid, naphthalenesulfonic acid, 1,5-naphthalenedisulfonic acid, camphoric acid, camphorsulfonic acid, salicylic acid, acetylsalicylic acid, aspartic acid, glutamic acid, lactic acid, gluconic acid, vitamin C acid, gallic acid, mandelic acid, malic acid, sorbic acid, trifluoroacetic acid, taurine, homotaurine, 2-hydroxyethanesulfonic acid and cinnamic acid.

[0040] The present invention also provides a pharmaceutical composition, the active ingredients of which include the TRPA1 modulators represented by the above formulas (I)-(VII) and pharmaceutically acceptable salts thereof and pharmaceutically acceptable excipients.

[0041] The present invention also provides the use of the TRPA1 regulator and the pharmaceutical composition as a TRPA1 regulator.

[0042] The present invention also provides an application of the above compound and pharmaceutical composition in preparing medicines for treating neurological diseases.

[0043] Furthermore, the drug for treating neurological diseases includes a drug for neuropathic pain.

[0044] In general, the above technical solution conceived by the present invention has the following technical advantages compared with the prior art:

[0045] 1. The present invention provides a TRPA1 ion channel modulator and its pharmaceutical composition and application. The TRPA1 ion channel modulator has high activity and greatly improved drug efficacy. Compared with the reported TRPA1 modulators AITC and GEN551, it has no obvious irritation and has good application prospects.

[0046] 2. The TRPA1 modulators provided by the present invention can quickly desensitize TRPA1 currents and do not have the irritation of reported TRPA1 modulators such as AITC and GEN551. Specifically, this type of compound does not cause irritation when injected into the hind paw of mice, and exhibits a good analgesic effect in a mouse pain model after oral administration. BRIEF DESCRIPTION OF THE DRAWINGS

[0047] Figure 1 Compound 1-40 (100 μM, corresponding to Figure 1 Abscissa number) Activation fold of TRPA1 current (normalized to positive control AITC).

[0048] Figure 2 This is the desensitization effect of compound 2 prepared in the example of the present invention on TRPA1 current.

[0049] Figure 3 The analgesic effect of compound 2 prepared in the example of the present invention on the mouse hot plate model.

[0050] Figure 4 The analgesic effect of compound 2 prepared in the example of the present invention on the CFA inflammatory pain model in mice.

[0051] Figure 5 The analgesic effect of compound 2 prepared in the example of the present invention on the mouse acetic acid model.

[0052] Figure 6 This is an irritation test on mice of compound 2 prepared in the example of the present invention. DETAILED DESCRIPTION

[0053] In order to make the purpose, technical scheme and advantages of the present invention clearer, the present invention is further described in detail below in conjunction with the embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not intended to limit the present invention. In addition, the technical features involved in the various embodiments of the present invention described below can be combined with each other as long as they do not conflict with each other.

[0054] Relevant terms of the present invention:

[0055] The hydrogen described in the present invention includes various isotopes of hydrogen, such as deuterium and tritium;

[0056] The term "hydrocarbon" (whether used alone or as part of another group) refers to any structure containing carbon atoms and hydrogen atoms;

[0057] The term "alkyl" refers to a linear or branched saturated hydrocarbon group, wherein the alkyl group may be optionally substituted with one or more substituents, including but not limited to methyl, ethyl, propyl (including all isomeric forms), n-propyl, isopropyl, butyl (including all isomeric forms), n-butyl, isobutyl, tert-butyl;

[0058] The term "alkoxy" is used to refer to an -O-alkyl group;

[0059] The term "aryl" refers to a monocyclic aromatic group and / or a polycyclic monovalent aromatic group containing at least one aromatic hydrocarbon ring; the aromatic group includes, but is not limited to, phenyl and biphenyl;

[0060] The term "heteroaryl" refers to a monocyclic heteroaryl containing heteroatoms N, O, S and / or a polycyclic monovalent heteroaryl containing at least one aromatic hydrocarbon ring; the heteroaryl includes but is not limited to thiophene, furan, pyrrole;

[0061] The term "halogen" is used to refer to fluorine, chlorine, bromine and iodine;

[0062] The term "pharmaceutically acceptable salt" refers to a salt prepared from a pharmaceutically acceptable non-toxic acid, including inorganic and organic acids. Suitable non-toxic acids include, but are not limited to, inorganic and organic acids, such as maleic acid, succinic acid, citric acid, tartaric acid, fumaric acid, formic acid, acetic acid, propionic acid, malonic acid, oxalic acid, benzoic acid, phthalic acid, methanesulfonic acid, benzenesulfonic acid, toluenesulfonic acid, naphthalenesulfonic acid, 1,5-naphthalenedisulfonic acid, camphoric acid, camphorsulfonic acid, salicylic acid, acetylsalicylic acid, aspartic acid, glutamic acid, lactic acid, gluconic acid, vitamin C acid, gallic acid, mandelic acid, malic acid, sorbic acid, trifluoroacetic acid, taurine, homotaurine, 2-hydroxyethanesulfonic acid, cinnamic acid, hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, nitric acid, phosphoric acid, and perchloric acid.

[0063] The instruments used for the experiments and sample analysis in the examples of this application are:

[0064] 1 HMR was measured on a Varian Mercuryplus 400 NMR spectrometer. 13 CMR was measured on a Bruker 600 MHz NMR instrument;

[0065] LS-MS was measured by Agilent 1100 Series LC / MSD1946D mass spectrometer;

[0066] Special chemical reagents were purchased from reagent companies such as Exploration, Bidex, Anaiji, Sigma, Alfa, Acros and Adamas. General chemical reagents were purchased from Sinopharm Shanghai Chemical Reagent Company. The main solvents included petroleum ether, ethyl acetate, dichloromethane, methanol, etc., which were purchased from Sinopharm Group and were all synthetic grade. Column chromatography silica gel used chemically pure silica gel from Huanghai Chemical Plant, generally 200-300 mesh. The reagent treatment method was treated with reference to Purification of Laboratory Chemicals (1988 edition).

[0067] The technical solution of the present invention is further described in detail below in conjunction with specific implementation methods.

[0068] Example 1

[0069] This embodiment provides a compound for preparing 4-(4-methoxyphenyl)-N-(3-methoxypropyl)thiazole-2-amine, and its general structural formula is:

[0070]

[0071] The reaction route is:

[0072]

[0073] The reaction steps are:

[0074] Compound 1 (2.5 g, 1 eq) was dissolved in ethanol (20 mL), thiourea (0.86 g, 1 eq) was added, and the mixture was refluxed for 2 h. After concentration under reduced pressure, water / saturated sodium bicarbonate (30 mL / 70 mL) was added and washed for 1 h. After the reaction was completed, the crude product compound 21.9 g was obtained by filtration, with a yield of 84%.

[0075] Compound 2 (176 mg, 1 eq), 3-methoxypropionic acid (133 mg, 1.5 eq), DIPEA (N, N-diisopropylethylamine, 165 mg, 1.5 eq) were dissolved in 5 mL DMF, and HCTU (6-chlorobenzotriazole-1,1,3,3-tetramethyluronium hexafluorophosphate, 530 mg, 1.5 eq) was added, and the mixture was reacted at room temperature for 24 h. After the reaction, saturated sodium bicarbonate, dilute hydrochloric acid, water, and saturated brine were added once for washing, dried over anhydrous sodium sulfate, and column chromatography with petroleum ether: ethyl acetate (3:1) was performed to obtain 3124 mg of the intermediate compound with a yield of 50%.

[0076] Lithium tetrahydrogen aluminum (0.68 mL, 2.5 M, 4 eq) was dissolved in anhydrous tetrahydrofuran, and the THF solution of compound 3 was added dropwise in an ice bath under argon protection, and the reaction was carried out in an ice bath for 6 h. After the reaction, 0.35 mL of water, 0.35 mL of sodium hydroxide (15% w / v) solution, 1.05 mL of water, and ethyl acetate were added, and the mixture was extracted with ethyl acetate, washed with saturated sodium bicarbonate and saturated brine, dried over anhydrous sodium sulfate, and chromatographed on a petroleum ether:ethyl acetate (3:1) column to obtain 51 mg of the final product 4-(4-methoxyphenyl)-N-(3-methoxypropyl)thiazole-2-amine, with a yield of 47%. 1 H NMR (600MHz, CDCl3) δ7.65(d,J=8.7Hz,2H),6.83(d,J=8.7Hz,2H),6.47(s,1H),5.69(s,1H) ,3.75(s,3H),3.44(t,J=5.8Hz,2H),3.33(d,J=4.3Hz,2H),3.28(s,3H),1.89-1.82(m,2H). 13 C NMR (151MHz, CDCl3) δ169.67,159.25,151.11,127.93,127.32,113.90,98.76,71.02,58.84,55.31,44.20,29.14.MS m / z:[M+H] + Calculate for C 14 H 18 N2O2S,279.1162; found,279.1170.

[0077] Example 1-1

[0078] Using Example 1

[0079] Similar operations were performed to obtain the following compound 2-40:

[0080]

[0081]

[0082]

[0083]

[0084]

[0085]

[0086]

[0087]

[0088] The electrophysiological experiments of the compounds prepared in the above examples are as follows:

[0089] The whole-cell patch clamp was used to record the activation effect of compound 1-40 on the current of TRPA1 channel. The patch clamp recording system used MultiClamp700A amplifier, 1440A digital-to-analog converter and pClamp10.6 acquisition and analysis software (Molecular devices, USA). The microelectrode was made of borosilicate hard glass electrode. The glass electrode was pulled by a puller (P-97, Sutter), and the electrode resistance was between 2-5MΩ after filling the electrode liquid. The cell was placed on the microscope stage, and the cell was selected to have a clear field of view. The pulled microelectrode was fixed on the amplifier probe, and the electrode was put into the liquid by a microelectrode manipulator, and slowly approached the cell. After the electrode contacted the cell, negative pressure was applied to form a GΩ seal between the electrode tip and the cell membrane surface. After compensating the electrode capacitance, negative pressure was applied to break the cell membrane, and the series resistance and cell membrane capacitance were compensated. Electrophysiological recording was performed using 6KHz filtering.

[0090] The results of electrophysiological experiments showed that the synthesized compounds can activate ( Figure 1 ) and desensitization ( Figure 2 )TRPA1 channels, and the effects of some compounds were significantly stronger than the known TRPA1 agonist AITC.

[0091] The animal experiments of the compounds prepared in the above examples are as follows:

[0092] Mouse hot plate model. C57BL / 6J female mice were used for the heat pain test, and the heat pain threshold of mice was determined by the hot plate method. Before the formal experiment began, the experimental mice were pre-screened and mice that were too sensitive or insensitive to the hot plate were shaved. The normal mouse withdrawal latency is 10-30 seconds, so mice with a withdrawal latency of less than 10 seconds or more than 30 seconds will be eliminated, and the interval between two measurement experiments must be greater than 10 minutes. The mice were randomly divided into 5 groups, 8 in each group, namely the blank control group (10% DMSO, 10% Tween 80 and 80% saline), compound 2 group (10, 20, 40 mg / kg) and positive drug ibuprofen group (20 mg / kg). 30 minutes before the start of the experiment, each group of mice was gavaged with the corresponding dose of compound. The withdrawal latency was calculated from the time the mouse was placed on the hot plate and terminated when the mouse was observed to lick or tap its paw quickly. The temperature of the metal hot plate was set to 55±0.5°C, and the paw withdrawal threshold of the mice was recorded before administration and at 0, 0.5, 1, 2, and 4 h after administration.

[0093] Acetic acid writhing model in mice. As with the hot plate experiment described above, C57BL / 6J male mice were randomly divided into five corresponding groups, with 8 mice in each group. Each group was intraperitoneally injected with 0.8% (v / v) acetic acid 30 minutes after administration. The blank control group was orally administered with solvent (10% DMSO, 10% Tween 80 and 80% saline), compound 2 (10, 20, 40 mg / kg) and the positive control ibuprofen group (20 mg / kg). After intraperitoneal injection of the corresponding dose of acetic acid (20 g / 200 μL) according to the weight of the mice, the mice were immediately placed in a transparent open cage. Abdominal contraction and hind limb extension were used as indicators of pain response. The cumulative number of abdominal contractions of the mice within 15 minutes was recorded.

[0094] Complete Freund's adjuvant-induced inflammatory pain model in mice. (1) Up and Down method to determine the basic mechanical withdrawal threshold of mice: C57BL / 6J male mice were first placed in a transparent glass container on a metal mesh and allowed to adapt to the container for 1 hour. After the adaptation, VonFrey hair of different strengths were used for the experiment, which were 2.0, 1.4, 1.0, 0.8, 0.6, 0.4, 0.16, 0.07, 0.04, and 0.02 (unit: g). The log units corresponding to different grams were 4.31, 4.17, 4.08, 3.84, 3.61, 3.22, 2.83, 2.44, 2.36, and 1.65, respectively. The mouse hind foot is stimulated according to the Up and Down method. Specifically, 0.4g of cilia are selected as the starting point. The cilia are in vertical contact with the hairless part of the palm and plantar part of the mouse's hind foot. The Von Frey hair is in an "S" shape and lasts for at least 2-3 seconds. When the mouse quickly lifts the tested foot, licks or bites the tested foot, it is a positive result, recorded as "X". If the above behavior does not occur, it is a negative result, recorded as "O". When a negative result appears, the next level of cilia with a stronger force should be used to stimulate the mouse's hind foot. When a positive result appears, the next level of cilia should be used to stimulate the mouse's hind foot. The interval between adjacent levels of cilia stimulation should be at least 10 seconds. After the first appearance of "OX" or "XO", continue to measure 4 times to obtain a series of XO values. According to the sequence results, refer to the k value table and determine the k value. Substitute the result value into the formula 50% g threshold = (10 [Xf+kδ]) / 10000, where δ = 0.224, Xf = the logarithm of the g value corresponding to the last measured cilium, and the 50% mechanical withdrawal threshold (50% MWT) of mice was calculated. Mice with outliers in the pain threshold were eliminated, and the remaining mice were used for the experiment. (2) Complete Freund's adjuvant (CFA)-induced symptomatic pain model in mice was established: the mice were placed in a prone position, the hind feet of the mice were fixed, and 20 μL of CFA solution was subcutaneously injected into the soles of the hind feet of the mice in the model group with an insulin syringe. 24 hours after the injection of CFA solution, the 50% mechanical withdrawal threshold of the hind feet of the mice in the model group was measured by Von Frey hair according to the Up and Down method, and the obtained value was compared with the basic mechanical withdrawal threshold. The mice with small differences in values ​​and failed modeling were eliminated, and the remaining mice were selected for formal experiments. (3) Up and Down method to detect the mechanical withdrawal threshold of mice: A total of five groups were set up in the experiment. The mice injected with CFA solution were randomly divided into a model control group (CFA group), a compound 2 group (10, 20, 40 mg / kg), and a positive control ibuprofen group (20 mg / kg), with 8 animals in each group. 30 minutes before the start of the formal experiment, each group of mice was gavaged with the corresponding solution according to their body weight, and the model control group was gavaged with 10% DMSO, 10% Tween 80 and 80% saline. The mechanical withdrawal threshold of mice was measured by Von Frey hair according to the Up and Down method before administration and at 0.5h, 1h, 2h, and 4h after administration.

[0095] Mouse irritation model. The compound was injected subcutaneously into the animal's hind paw. The irritation of the drug was manifested by the animal's vigorous licking and biting of the injected paw. According to different groups, 20 μL of solvent (10% DMSO, 10% Tween 80 and 80% saline, blank control group), AITC (1%) and compound 2 (1%) were subcutaneously injected into the hind paw of C57BL / 6J male mice. Then the mice were immediately placed in a plastic observation chamber, and the licking and biting of the injected paw by the mice was observed and recorded. The total time that the mice licked and bit the injected paw was recorded in units of 5 minutes, for a total of 45 minutes.

[0096] Animal experiment results show that compound 2 has a good analgesic effect in animals ( Figure 3-5 ), and no obvious irritation at high doses ( Figure 6 ).

[0097] It will be easily understood by those skilled in the art that 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 and improvements made within the spirit and principles of the present invention should be included in the protection scope of the present invention.

Claims

1. A TRPA1 modulator, characterized in that Its general structural formula is shown in Formula I: wherein Ring A is selected from C6-12 aryl or 5-10 membered heteroaryl, wherein the aryl or heteroaryl is optionally substituted with 0-3 halogen, amino, hydroxy, carboxyl, nitro, trifluoromethyl, amide, methoxy, cyano or C1-4 alkyl; R1 is selected from one of hydrogen, halogen, cyano, nitro, amino, hydroxyl, carboxyl, trifluoromethyl, amide, methoxy, C1-C4 alkyl, C1-C4 alkoxy, and C3-C6 cycloalkyl; R2 and R3 are selected from H or C1-C6 alkyl, or C1-C6 alkyl substituted by hydroxy, amino, C1-C4 alkoxy, C1-C4 alkylcarbonyl, halogen atom, C6-10 aryl or halogenated C6-10 aryl, C3-C6 cycloalkyl, C1-C6 alkylcarbonyl, C1-C6 alkoxycarbonyl, C1-C6 alkylaminocarbonyl or C2-C8 alkenyl; or C2-C8 alkenyl, C5-C7 cycloalkenyl or C2-C8 alkynyl substituted by a C1-C4 alkylcarbonyl, a C1-C4 alkoxy, a halogen atom, a C6-C10 aryl or a halogenated C6-C10 aryl group; or C2-C8 alkynyl or C6-C10 aryl substituted by a hydroxyl, an amino, a C1-C4 alkylcarbonyl, a C1-C4 alkoxy, a halogen atom, a C6-C10 aryl or a halogenated C6-C10 aryl group.

2. The TRPA1 modulator according to claim 1, characterized in that The general structural formula of the TRPA1 modulator is shown in Formula II: Wherein, R1 is selected from one of hydrogen, halogen, cyano, nitro, amino, hydroxyl, carboxyl, trifluoromethyl, amide, methoxy, C1-C4 alkyl, C1-C4 alkoxy, and C3-C6 cycloalkyl; R2 and R3 are selected from H or C1-C6 alkyl, or C1-C6 alkyl substituted by hydroxyl, amino, C1-C4 alkoxy, C1-C4 alkylcarbonyl, halogen atom, C6-10 aryl or halogenated C6-10 aryl, C3-C6 cycloalkyl, C1-C6 alkylcarbonyl, C1-C6 alkoxycarbonyl, C1-C6 alkylaminocarbonyl or C2-C8 alkenyl, or C1-C6 alkyl substituted by hydroxyl, amino, C1-C4 alkoxy, C1-C4 alkylcarbonyl, halogen atom, C6-10 aryl or halogenated C6-10 aryl. C2-C8 alkenyl, C5-C7 cycloalkenyl or C2-C8 alkynyl substituted by 4-alkylcarbonyl, C1-C4 alkoxy, halogen atom, C6-C10 aryl or halogenated C6-C10 aryl, or C2-C8 alkynyl or C6-C10 aryl substituted by hydroxy, amino, C1-C4 alkylcarbonyl, C1-C4 alkoxy, halogen atom, C6-C10 aryl or halogenated C6-C10 aryl; Wherein, X and Y are each independently selected from C, O, N or S.

3. The TRPA1 modulator according to claim 2, characterized in that The general structural formula of the TRPA1 modulator is shown in Formula III: wherein R1 is selected from hydrogen, halogen, cyano, nitro, amino, hydroxyl, carboxyl, trifluoromethyl, amide, methoxy, C1-C4 alkyl, C1-C4 alkoxy or C3-C6 cycloalkyl; R2 and R3 are selected from H or C1-C6 alkyl, or C1-C6 alkyl substituted by hydroxyl, amino, C1-C4 alkoxy, C1-C4 alkylcarbonyl, halogen atom, C6-10 aryl or halogenated C6-10 aryl, C3-C6 cycloalkyl, C1-C6 alkylcarbonyl, C1-C6 alkoxycarbonyl, C1-C6 alkylaminocarbonyl or C2-C8 alkenyl, or C1-C6 alkyl substituted by hydroxyl, amino, C1-C4 alkoxy, C1-C4 alkylcarbonyl, halogen atom, C6-10 aryl or halogenated C6-10 aryl. The invention also comprises a C2-C8 alkenyl, C5-C7 cycloalkenyl or C2-C8 alkynyl group substituted with a C1-C4 alkylcarbonyl group, a C1-C4 alkoxy group, a halogen atom, a C6-C10 aryl group or a halogenated C6-C10 aryl group, or a C2-C8 alkynyl or C6-C10 aryl group substituted with a hydroxyl group, an amino group, a C1-C4 alkylcarbonyl group, a C1-C4 alkoxy group, a halogen atom, a C6-C10 aryl group or a halogenated C6-C10 aryl group.

4. The TRPA1 modulator according to claim 2, characterized in that The general structural formula of the TRPA1 modulator is shown in Formula IV: wherein R1 is selected from hydrogen, halogen, cyano, nitro, amino, hydroxyl, carboxyl, trifluoromethyl, amide, methoxy, C1-C4 alkyl, C1-C4 alkoxy or C3-C6 cycloalkyl; R2 is selected from H or C1-C6 alkyl, or selected from C1-C6 alkyl substituted by hydroxyl, amino, C1-C4 alkoxy, C1-C4 alkylcarbonyl, halogen atom, C6-10 aryl or halogenated C6-10 aryl, C3-C6 cycloalkyl, C1-C6 alkylcarbonyl, C1-C6 alkoxycarbonyl, C1-C6 alkylaminocarbonyl or C2-C8 alkenyl, or selected from C1-C6 alkyl substituted by hydroxyl, amino, C1-C4 alkoxy, C1-C4 alkylcarbonyl, halogen atom, C6-10 aryl or halogenated C6-10 aryl C2-C8 alkenyl, C5-C7 cycloalkenyl or C2-C8 alkynyl substituted by alkylcarbonyl, C1-C4 alkoxy, halogen atom, C6-C10 aryl or halogenated C6-C10 aryl, or selected from C2-C8 alkynyl or C6-C10 aryl substituted by hydroxyl, amino, C1-C4 alkylcarbonyl, C1-C4 alkoxy, halogen atom, C6-C10 aryl or halogenated C6-C10 aryl.

5. The TRPA1 modulator according to claim 2, characterized in that The general structural formula of the TRPA1 modulator is shown in Formula V: wherein R1 is selected from hydrogen, halogen, cyano, nitro, amino, hydroxyl, carboxyl, trifluoromethyl, amide, methoxy, C1-C4 alkyl, C1-C4 alkoxy or C3-C6 cycloalkyl; R2 is selected from H or C1-C6 alkyl, or selected from C1-C6 alkyl substituted by hydroxyl, amino, C1-C4 alkoxy, C1-C4 alkylcarbonyl, halogen atom, C6-10 aryl or halogenated C6-10 aryl, C3-C6 cycloalkyl, C1-C6 alkylcarbonyl, C1-C6 alkoxycarbonyl, C1-C6 alkylaminocarbonyl or C2-C8 alkenyl, or selected from C1-C6 alkyl substituted by hydroxyl, amino, C1-C4 alkoxy, C1-C4 alkylcarbonyl, halogen atom, C6-10 aryl or halogenated C6-10 aryl C2-C8 alkenyl, C5-C7 cycloalkenyl or C2-C8 alkynyl substituted by alkylcarbonyl, C1-C4 alkoxy, halogen atom, C6-C10 aryl or halogenated C6-C10 aryl, or selected from C2-C8 alkynyl or C6-C10 aryl substituted by hydroxyl, amino, C1-C4 alkylcarbonyl, C1-C4 alkoxy, halogen atom, C6-C10 aryl or halogenated C6-C10 aryl.

6. The TRPA1 modulator according to claim 2, characterized in that The general structural formula of the TRPA1 modulator is shown in Formula VI: wherein R1 is selected from hydrogen, halogen, cyano, nitro, amino, hydroxyl, carboxyl, trifluoromethyl, amide, methoxy, C1-C4 alkyl, C1-C4 alkoxy or C3-C6 cycloalkyl; R2 is selected from H or C1-C6 alkyl, or selected from C1-C6 alkyl substituted by hydroxyl, amino, C1-C4 alkoxy, C1-C4 alkylcarbonyl, halogen atom, C6-10 aryl or halogenated C6-10 aryl, C3-C6 cycloalkyl, C1-C6 alkylcarbonyl, C1-C6 alkoxycarbonyl, C1-C6 alkylaminocarbonyl or C2-C8 alkenyl, or selected from C1-C6 alkyl substituted by hydroxyl, amino, C1-C4 alkoxy, C1-C4 alkylcarbonyl, halogen atom, C6-10 aryl or halogenated C6-10 aryl C2-C8 alkenyl, C5-C7 cycloalkenyl or C2-C8 alkynyl substituted by alkylcarbonyl, C1-C4 alkoxy, halogen atom, C6-C10 aryl or halogenated C6-C10 aryl, or selected from C2-C8 alkynyl or C6-C10 aryl substituted by hydroxyl, amino, C1-C4 alkylcarbonyl, C1-C4 alkoxy, halogen atom, C6-C10 aryl or halogenated C6-C10 aryl.

7. The TRPA1 modulator according to claim 2, characterized in that The general structural formula of the TRPA1 modulator is shown in Formula VII: wherein R1 is selected from hydrogen, halogen, cyano, nitro, amino, hydroxyl, carboxyl, trifluoromethyl, amide, methoxy, C1-C4 alkyl, C1-C4 alkoxy or C3-C6 cycloalkyl; R2 is selected from H or C1-C6 alkyl, or selected from C1-C6 alkyl substituted by hydroxyl, amino, C1-C4 alkoxy, C1-C4 alkylcarbonyl, halogen atom, C6-10 aryl or halogenated C6-10 aryl, C3-C6 cycloalkyl, C1-C6 alkylcarbonyl, C1-C6 alkoxycarbonyl, C1-C6 alkylaminocarbonyl or C2-C8 alkenyl, or selected from C1-C6 alkyl substituted by hydroxyl, amino, C1-C4 alkoxy, C1-C4 alkylcarbonyl, halogen atom, C6-10 aryl or halogenated C6-10 aryl C2-C8 alkenyl, C5-C7 cycloalkenyl or C2-C8 alkynyl substituted by alkylcarbonyl, C1-C4 alkoxy, halogen atom, C6-C10 aryl or halogenated C6-C10 aryl, or selected from C2-C8 alkynyl or C6-C10 aryl substituted by hydroxyl, amino, C1-C4 alkylcarbonyl, C1-C4 alkoxy, halogen atom, C6-C10 aryl or halogenated C6-C10 aryl.

8. The TRPA1 modulator according to claim 2, characterized in that The TRPA1 modulator is selected from the following compounds:

9. A pharmaceutical composition, characterized in that The active ingredients of the pharmaceutical composition include the TRPA1 modulator according to any one of claims 1 to 8, and its stereoisomers, prodrugs, solvates, hydrates, and pharmaceutically acceptable inorganic acids or organic acid salts thereof; The inorganic acid includes any one or more of hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfonic acid, carbonic acid, sulfuric acid, phosphoric acid, nitric acid and perchloric acid; The organic acid includes any one or more of maleic acid, succinic acid, citric acid, tartaric acid, fumaric acid, formic acid, acetic acid, propionic acid, malonic acid, oxalic acid, benzoic acid, phthalic acid, methanesulfonic acid, benzenesulfonic acid, toluenesulfonic acid, naphthalenesulfonic acid, 1,5-naphthalenedisulfonic acid, camphoric acid, camphorsulfonic acid, salicylic acid, acetylsalicylic acid, aspartic acid, glutamic acid, lactic acid, gluconic acid, vitamin C acid, gallic acid, mandelic acid, malic acid, sorbic acid, trifluoroacetic acid, taurine, homotaurine, 2-hydroxyethanesulfonic acid and cinnamic acid.

10. Use of the TRPA1 modulator according to any one of claims 1 to 8 or the pharmaceutical composition according to claim 9 in the preparation of a medicament for treating neurological diseases.

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