2-substituted aminobenzothiazole compounds and uses thereof

Abstract

The present application relates to 2-substituted aminobenzothiazole compounds and pharmaceutically acceptable salts thereof, and pharmaceutical compositions containing the compounds and uses thereof. The 2-substituted aminobenzothiazole compounds are compounds of general formula I and pharmaceutically acceptable salts thereof, wherein the substituents R 1 , R 2 , R 3 , R 4 , m, n, X, Y have the meanings given in the description. The present application also relates to the use of the compounds of general formula I for the preparation of a medicament for the treatment and / or prevention of sEH-mediated diseases, in particular for the preparation of a medicament for the treatment of inflammatory diseases, cardiovascular and cerebrovascular diseases, pain, dry eye, diabetes, diabetic complications, diabetes-related diseases, fibrotic diseases, neurological and psychiatric diseases, ulcerative diseases, etc.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of pharmaceutical chemistry, in particular to a series of 2-substituted aminobenzothiazole compounds, the preparation method and the use thereof in the preparation of drugs for treating and / or preventing sEH-mediated diseases. BACKGROUND

[0002] Epoxide hydrolase (EH) is widely present in mammals, plants and microorganisms in nature, and can efficiently catalyze the hydrolysis of epoxide to generate vicinal diols. The epoxide hydrolases found include 8 subtypes of cholesterol 5,6-oxide hydrolase (ChEH), soluble epoxide hydrolase (sEH) and microsomal epoxide hydrolase (mEH). Among them, sEH is one of the epoxide hydrolases that has been studied in depth in recent years, and sEH is a potential therapeutic target for many diseases such as hypertension, lung disease, diabetes, pain, inflammation and other immune system diseases.

[0003] sEH is encoded by EPHX2 gene, which is located in the P21-P12 region of human chromosome 8, including 18 introns and 19 exons, and encodes more than 550 amino acids with a total length of 45 kb. sEH is composed of two 60kD subunits in an anti-parallel manner, and each subunit contains two different enzyme active domains. The C-terminal has an α / β hydrolytic enzyme folding structure, which can hydrolyze epoxide to form the corresponding diol as a catalytic subunit; the N-terminal has phosphatase activity, which can hydrolyze various lipid phosphates. sEH is widely distributed in the brain, liver, kidney and many other organs of the human body, and has a lower content in the lung, spleen and other organs. In recent years, sEH has also been found in the cortex, hippocampus, amygdala and striatum of the brain.

[0004] Arachidonic acid (AA) is an unsaturated fatty acid, which can be metabolized by cyclooxygenase (COX) and lipoxygenase (LOX) into inflammatory mediators such as prostaglandins, lipoxins and leukotrienes. AA can be converted into epoxyeicosatrienoic acids (EETs) with higher biological activity by cytochrome P450 (CYP450) enzymes. EETs have anti-inflammatory, anti-apoptotic, antioxidant and other effects, and are involved in the pathophysiological processes of various diseases of the immune system, nervous system and metabolic system. sEH is the main metabolic enzyme of EETs, which can rapidly hydrolyze EETs into dihydroxyeicosatrienoic acids (DHETs) with lower biological activity, affecting the protective effect of EETs. sEH inhibitors can increase the level of endogenous EETs, maintain the biological activity of EETs, and thus achieve the purpose of treating various diseases.

[0005] Currently, there is no sEH inhibitor applied in clinic. Literature reports that sEH inhibitors include t-AUCB, AR9281, GSK2256294 and EC5026. GSK2256294 is a small molecule sEH inhibitor discovered by GlaxoSmithKline, which can be used for treating diseases such as diabetes and obesity, and is currently in phase II clinical study. EC5026 is a urea sEH inhibitor discovered by Bruce Hammock team, and the compound is currently in phase I clinical study.

[0006] SUMMARY

[0007] The purpose of the present application is to provide a 2-substituted aminobenzothiazole compound, which is suitable for preparing a drug for treating and / or preventing sEH-mediated diseases.

[0008] To achieve the above purpose, the technical scheme adopted by the present application is as follows:

[0009] The present application relates to a 2-substituted aminobenzothiazole compound and a pharmaceutically acceptable salt thereof as shown in general formula I,

[0010]

[0011] Among them,

[0012] When Y represents an N atom, X represents an S atom; when Y represents an S atom, X represents an N atom;

[0013] R 1 and R2 are independently selected from hydrogen, (C1-C6)alkyl, (C3-C7)cycloalkyl; or R 1 and R 2 together with the nitrogen atom to which they are attached form a 4-10 membered heterocyclyl group, which heterocyclyl group optionally contains, in addition to the nitrogen atom, 0-3 heteroatoms selected from N, O, or S, and which heterocyclyl group is optionally substituted with 1-3 R 5 groups, which heterocyclyl group contains 0-2 carbon-carbon double bonds;

[0014] R 5 is a hydrogen atom, =0, hydroxy, amino, halogen, carboxy, cyano, nitro, (C1-C6)alkyl, (C1-C6)alkoxy, (C3-C7)cycloalkyl, (C1-C6)alkylsulfinyl, (C1-C6)alkylsulfonyl, or (C1-C6)alkylformyl;

[0015] R 3 is a (C6-C 10 )aryl, 5-10 membered heteroaryl, or adamantyl group, which heteroaryl group contains 1-3 heteroatoms selected from N, O, or S, and which aryl or heteroaryl group is optionally substituted with 1-3 R 6 groups, which are the same or different;

[0016] R 6 is a hydrogen atom, hydroxy, halogen, cyano, mercapto, carboxy, carbamoyl, substituted or unsubstituted (C1-C6)alkyl, substituted or unsubstituted (C1-C6)alkoxy, substituted or unsubstituted (C1-C6)alkylthio, substituted or unsubstituted (C2-C6)alkenyl, substituted or unsubstituted (C1-C6)alkylsulfinyl, substituted or unsubstituted (C1-C6)alkylsulfonyl, substituted or unsubstituted (C1-C6)alkylformyl, substituted or unsubstituted (C1-C6)alkoxycarbonyl, -CO-NHR 7 , -CO-NR 7 R 8 , -NH-CO-R 7 , -NHR 7 , or -NR 7 R 8 ; wherein R 7 , R 8 are the same or different substituted or unsubstituted (C1-C6)alkyl; each of the above-mentioned substituents is selected from hydroxy, amino, or halogen; the substitution is mono- or polysubstitution, and the substituents in the polysubstitution are the same or different;

[0017] R 4 is hydrogen, halogen, (C1-C4)alkyl, (C1-C4)alkoxy;

[0018] m is 0, 1, or 2;

[0019] n is 2, 3 or 4.

[0020] The present application preferably relates to 2-substituted aminobenzothiazoles of the general formula I

[0021] wherein

[0022] X represents a S atom when Y represents a N atom; X represents a N atom when Y represents a S atom;

[0023] R 1 and R 2 together with the nitrogen atom to which they are attached form a 4-10 membered heterocyclyl group, which heterocyclyl group optionally contains, in addition to the nitrogen atom, 0-3 heteroatoms selected from N, O or S, which heterocyclyl group can also optionally be substituted with 1-3 R 5 groups which are the same or different; the heterocyclyl group contains 0-2 carbon-carbon double bonds;

[0024] R 5 is a hydrogen atom, =0, hydroxy, amino, halogen, carboxy, cyano, nitro, (C1-C4)alkyl, (C1-C4)alkoxy, (C3-C6)cycloalkyl, (C1-C4)alkylsulfinyl, (C1-C4)alkylsulfonyl, (C1-C4)alkylformyl;

[0025] R 3 is a phenyl group, a 5-6 membered heteroaryl group or an adamantyl group, the heteroaryl group containing 1-3 heteroatoms selected from N, O or S, which phenyl or heteroaryl group can also optionally be substituted with 1-3 R 6 groups which are the same or different;

[0026] R 6 is a hydrogen atom, hydroxy, halogen, cyano, mercapto, carboxy, carbamoyl, substituted or unsubstituted (C1-C4)alkyl, substituted or unsubstituted (C1-C4)alkoxy, substituted or unsubstituted (C1-C4)alkylthio, substituted or unsubstituted (C2-C4)alkenyl, substituted or unsubstituted (C1-C4)alkylsulfinyl, substituted or unsubstituted (C1-C4)alkylsulfonyl, substituted or unsubstituted (C1-C4)alkylformyl, substituted or unsubstituted (C1-C4)alkoxycarbonyl, -CO-NHR 7 , -CO-NR 7 R 8 , -NH-CO-R 7 , -NHR 7 or -NR 7 R 8 ; wherein R 7 , R 8are identical or different, substituted or unsubstituted (C1-C4)alkyl; each of the above-mentioned substituents is selected from the group consisting of hydroxy, amino or halogen; the substitution is mono- or polysubstitution, the substituents in polysubstitution being identical or different;

[0027] R 4 is hydrogen, halogen or methyl;

[0028] m is 0 or 1;

[0029] n is 2 or 3.

[0030] The present application more preferably relates to 2-substituted aminobenzothiazole compounds of the general formula I

[0031] X represents a S atom when Y represents a N atom; X represents a N atom when Y represents a S atom;

[0032] R 1 and R 2 together with the nitrogen atom to which they are attached form

[0033]

[0034] R 3 is phenyl or adamantyl; phenyl is optionally substituted by 1 to 3 identical or different R 6 substituents;

[0035] R 6 is halogen, unsubstituted or optionally substituted (C1-C4)alkyl or (C1-C4)alkoxy by 1 to 3 identical or different halogen;

[0036] R 4 is hydrogen, fluorine or chlorine;

[0037] m is 0 or 1;

[0038] n is 2.

[0039] The present application more preferably relates to 2-substituted aminobenzothiazole compounds of the general formula I

[0040] X represents a S atom when Y represents a N atom; X represents a N atom when Y represents a S atom, and R 4 is H; R 1 and R 2 together with the nitrogen atom to which they are attached form

[0041]

[0042] R 3 is phenyl, the para position of which is substituted by R 6 ; and

[0043] R 6 is halogen, trifluoromethyl or trifluoromethoxy;

[0044] R 4 is hydrogen, fluorine or chlorine;

[0045] m is 0 or 1 ;

[0046] n is 2.

[0047] The present application particularly preferably relates to 2-substituted aminobenzothiazole compounds of the general formula II and pharmaceutically acceptable salts thereof, wherein

[0048]

[0049] R 1 and R 2 together with the nitrogen atom to which they are attached form

[0050]

[0051] R 3 is phenyl, which is substituted in the para position by R 6 ;

[0052] R 6 is halogen, trifluoromethyl or trifluoromethoxy;

[0053] R 4 is hydrogen, fluorine or chlorine;

[0054] m is 0 or 1 ;

[0055] n is 2.

[0056] More preferably, the present application proposes the following 2-substituted aminobenzothiazole compounds and pharmaceutically acceptable salts thereof:

[0057] 1-{N-[2-(4-morpholinyl)ethyl]-2-aminobenz[d]thiazol-5-yl}-3-(4-chlorophenyl)urea;

[0058] 1-{N-[2-(2-oxo-1-pyrrolidinyl)ethyl]-2-aminobenz[d]thiazol-5-yl}-3-(4-chlorophenyl)urea;

[0059] 1-{N-[2-(4-morpholinyl)ethyl]-2-aminobenz[d]thiazol-6-yl}-3-(4-chlorophenyl)urea;

[0060] 1-{N-[2-(dimethylamino)ethyl]-2-aminobenz[d]thiazol-6-yl}-3-(4-chlorophenyl)urea;

[0061] 1-{N-[2-(4-morpholinyl)ethyl]-2-amino-benzo[d]thiazol-5-yl}-3-(3-fluoro-4- chlorophenyl)urea;

[0062] 1-{N-[2-(1-piperidinyl)ethyl]-2-amino-benzo[d]thiazol-5-yl}-3-(4-chlorophenyl)urea;

[0063] 1-{N-[2-(4-morpholinyl)ethyl]-2-amino-benzo[d]thiazol-6-yl}-3-(4- trifluoromethoxyphenyl)urea;

[0064] 1-{N-[2-(2-oxo-1-pyrrolidinyl)ethyl]-2-amino-benzo[d]thiazol-5-yl}-3-(3-fluoro-4- chlorophenyl)urea;

[0065] 1-{N-[2-(4-morpholinyl)ethyl]-2-amino-benzo[d]thiazol-6-yl}-3-(1-adamantyl)urea;

[0066] 1-{N-[2-(2-oxo-1-pyrrolidinyl)ethyl]-2-amino-benzo[d]thiazol-6-yl}-3-(4- chlorophenyl)urea;

[0067] 1-{N-[2-(3-oxo-4-morpholinyl)ethyl]-2-amino-benzo[d]thiazol-5-yl}-3-(3,4- dichlorophenyl)urea;

[0068] 1-{N-[2-(3-oxo-4-morpholinyl)ethyl]-2-amino-5-fluoro-benzo[d]thiazol-6-yl}-3-(4- chlorophenyl)urea;

[0069] 1-{N-[2-(2-oxo-1-piperidinyl)ethyl]-2-amino-5-fluoro-benzo[d]thiazol-6-yl}-3-(4- chlorophenyl)urea;

[0070] 1-{N-[2-(3-oxo-4-morpholinyl)ethyl]-2-amino-benzo[d]thiazol-6-yl}-3-(1- adamantyl)urea;

[0071] 1-{N-[2-(3-oxo-4-morpholinyl)ethyl]-2-amino-5-chloro-benzo[d]thiazol-6-yl}-3-(4- chlorophenyl)urea;

[0072] 1-{N-[2-(2-oxo-1-pyrrolidinyl)ethyl]-2-amino-5-fluoro-benzo[d]thiazol-6-yl}-3-(4- chlorophenyl)urea.

[0073] Alternatively, the pharmaceutically acceptable salt of the 2-substituted aminobenzothiazole compound is a salt of the 2-substituted aminobenzothiazole compound with an acid selected from the group consisting of hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid, formic acid, acetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, picric acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, benzenesulfonic acid, naphthalenesulfonic acid, trifluoroacetic acid, and aspartic acid.

[0074] The present application also includes prodrugs of the compounds of the present application. Prodrugs of the compounds of the present application are compounds of Formula I which can have little or no pharmacological activity themselves, but which are converted to the corresponding biologically active form, e.g., by metabolic oxidation, solvolysis or some other mechanism, following administration.

[0075] Unless otherwise indicated, the term "halogen" as used herein means fluorine, chlorine or bromine. "Hydroxy" means -OH. "Amino" means -NH2. "Carboxy" means -COOH. "Cyano" means -CN. "Nitro" means -NO2. "Carbamoyl" means -CO-NH2.

[0076] "C1-C4" means that the group (e.g., alkyl, alkoxy, cycloalkyl, etc.) defined contains 1, 2, 3, or 4 carbon atoms. The meaning of other terms similarly described in a like manner, e.g., "C1-C6", "C3-C6", etc.

[0077] "Alkyl" means straight or branched chain alkyl. For example, "C1-C6alkyl" means straight or branched chain alkyl having 1, 2, 3, 4, 5, or 6 carbon atoms, and can be selected from the group consisting of methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl, t-butyl, pentyl, and hexyl. "Alkenyl" means straight or branched chain alkenyl.

[0078] "Cycloalkyl" means substituted or unsubstituted cycloalkyl. For example, "C3-C6cycloalkyl" means cycloalkyl having 3, 4, 5, or 6 carbon atoms, and can be selected from the group consisting of cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

[0079] "Alkoxy" is an -O-R group, "alkylthio" is an -S-R group, "alkylsulfmyl" is an -SO-R group, "alkylsulfonyl" is an -SO2-R group, "alkylcarbonyl" is a -CO-R group, and "alkoxycarbonyl" is a -CO-O-R group, where R is alkyl as defined above.

[0080] "Heteroaryl" means a monocyclic or polycyclic ring system containing one or more heteroatoms selected from N, O, S, which ring system is aromatic, such as imidazolyl, pyridyl, pyrazolyl, furanyl, thienyl, pyrrolyl, thiazolyl, benzothiazolyl, oxazolyl, isoxazolyl, quinolinyl, isoquinolinyl, benzimidazolyl, benzoxazolyl, and the like; "heterocyclyl" means a monocyclic or polycyclic ring system containing one or more heteroatoms selected from N, O, S, such as pyrrolidinyl, morpholinyl, piperazinyl, piperidinyl, pyrazolidinyl, imidazolidinyl, and thiazolinyl, and the like.

[0081] The present application also provides a pharmaceutical composition comprising one or more of the above-mentioned 2-substituted aminobenzothiazole compounds and pharmaceutically acceptable salts thereof and a pharmaceutically acceptable carrier. The compounds of the present application can be used in combination with other active ingredients as long as they do not produce adverse effects, such as allergic reactions.

[0082] The carriers used in the pharmaceutical composition of the present application are of the common types available in the pharmaceutical field, including: binders, lubricants, disintegrants, co-solvents, diluents, stabilizers, suspending agents, pigments, flavoring agents, and the like for oral preparations; pH adjusters, osmotic pressure adjusters, solubilizers, stabilizers, and the like for injectable preparations; bases, diluents, lubricants, preservatives, and the like for topical preparations. The pharmaceutical preparations can be administered orally, parenterally (e.g., intravenously, subcutaneously, intraperitoneally, and the like), or topically (e.g., ocularly, nasally, sublingually, dermally, and the like), and if certain drugs are not stable in the stomach, they can be formulated into enteric-coated tablets.

[0083] It is found through in vitro and in vivo inhibition activity tests that the compounds of general formula I and pharmaceutically acceptable salts thereof in the present application have significant inhibitory activity on sEH and can stabilize the level of EETs in vivo. The results of pharmacodynamic studies show that the compounds in the present application can inhibit carrageenan-induced paw swelling, protect against ischemic brain injury, maintain blood glucose homeostasis, relieve neuropathic pain, resist tissue fibrosis, relieve depression, improve dry eye symptoms, and the like, and can be used for treating sEH-mediated inflammatory diseases, cardiovascular and cerebrovascular diseases, diabetes and complications, fibrotic diseases, neurological and mental diseases, pain, ulcerative diseases, or dry eye, and the like.

[0084] The precise amount of the compounds of the present application required to treat a condition mediated by sEH will vary depending on the subject, the species, age, and general condition of the subject, the severity of the condition being treated, the particular compound used, and its mode of administration, e.g., the route and frequency of administration, and the like. An appropriate effective amount can be determined by one of ordinary skill in the art with only routine experimentation.

[0085] The amount of compound to be administered can vary from about 0.1 to 100 mg / kg body weight per day, preferably 1 to 50 mg / kg body weight per day. It will be appreciated that the dose will vary depending on the requirements of the patient, the severity of the inflammatory disease, cardiovascular disease, diabetes, diabetic complications, diabetes-related diseases, fibrotic diseases, neurological and psychiatric diseases, pain, ulcerative diseases or dry eye, and the particular compound being employed. Also, it will be appreciated that the initially administered dose can exceed the upper limit, with the dose being subsequently reduced, as quickly as possible, to the minimum effective level, or the initially administered dose can be less than the optimum, with the daily dosage being progressively increased during the course of treatment, depending on the specific circumstances. If desired, the daily dosage can be divided into multiple doses for administration, e.g., two to four times per day.

[0086] Mammal means human or animal.

[0087] The amount of active ingredient, i.e., the compound according to the present application, in the pharmaceutical composition and unit dosage form thereof can vary, depending on the particular application, the potency and the desired concentration of the particular compound. In general, the amount of active ingredient will be between 0.5% and 90% by weight of the total composition.

[0088] In combination therapy, the compound of the present application and the other compound can be administered simultaneously or sequentially, in which case the compound of the present application and the other compound can be combined in a single pharmaceutical composition or in separate compositions.

[0089] The present application also provides the use of a compound of Formula I and pharmaceutically acceptable salts or pharmaceutical compositions thereof for the manufacture of a medicament for the treatment and / or prevention of sEH-mediated diseases. The diseases include inflammatory diseases, cardiovascular diseases, diabetes, diabetic complications, diabetes-related diseases, fibrotic diseases, neurological and psychiatric diseases, pain, ulcerative diseases and dry eye.

[0090] Specifically, the inflammatory diseases include inflammatory liver diseases, inflammatory kidney diseases, inflammatory lung diseases, inflammatory brain diseases, myocarditis, pancreatitis, arthritis, soft tissue inflammation, bone tissue inflammation and vascular inflammation; the cardiovascular and cerebrovascular diseases include hypertension, myocardial infarction, heart failure, coronary heart disease, cardiovascular arteriosclerosis, ischemic stroke and hemorrhagic stroke; the diabetes includes type I diabetes or type II diabetes; the diabetes complications include diabetic retinopathy, diabetes-related uveitis, diabetic cataract, diabetic nephropathy, diabetic skin disease and diabetic peripheral neuropathy; the diabetes-related diseases include hyperlipidemia, hyperuricemia and gout, obesity and metabolic syndrome; the fibrosis diseases include pulmonary fibrosis, liver fibrosis, myocardial fibrosis and renal fibrosis; the neurological and mental diseases include Alzheimer's disease, epilepsy, Parkinson's disease, amyotrophic lateral sclerosis, schizophrenia, mental disorders, depression, neurasthenia; the pain diseases include neuropathic pain, inflammatory pain, oncological pain and mixed pain; the ulcer diseases include gastric ulcer, duodenal ulcer, ulcerative colitis, corneal ulcer and oral ulcer; the dry eye diseases include aqueous deficiency dry eye, lipid abnormality dry eye, mucin abnormality dry eye, tear dynamics abnormality dry eye and mixed dry eye.

[0091] The present application relates to a method for treating and / or preventing sEH-mediated diseases by administering a compound of Formula I and pharmaceutically acceptable salts thereof to a subject in need thereof. The compound of Formula I and pharmaceutically acceptable salts thereof of the present application show sEH inhibitor activity, and thus can be effectively used for preventing or treating sEH-mediated related diseases.

[0092] In the present application, the term "prevention" means any action resulting from the administration of the compound or composition of the present application to inhibit or delay the occurrence, spread and recurrence of sEH-mediated related diseases, and the term "treatment" means all actions to relieve or beneficially change the symptoms of the diseases by administering the compound or composition of the present application.

[0093] The examples and preparation examples provided below further illustrate and exemplify the compounds of the present application and methods for preparing them. It is to be understood that the scope of the application is not to be in any way limited by the following examples and preparation examples.

[0094] The following synthetic routes describe the preparation of the compounds of Formula I of the present application, all starting materials are prepared by the methods described in these synthetic routes, by methods well known to those of ordinary skill in the art of organic chemistry, or are commercially available. All final compounds of the present application are prepared by the methods described in these synthetic routes or by methods analogous thereto, which are well known to those of ordinary skill in the art of organic chemistry. All variable factors used in these synthetic routes are defined below or as defined in the claims.

[0095] The synthesis route of the compound of formula I is as follows: raw material A is reacted with Boc2(O) to obtain intermediate B, intermediate B is reacted with R 1 R 2 N(CH2) n Cl to obtain intermediate C by nucleophilic substitution reaction, intermediate C is reduced by iron acid or H2 / Pd-C to obtain intermediate D; intermediate D is reacted with intermediate E to obtain intermediate F by condensation reaction, and the hydrochloride salt of the compound of formula I is obtained after intermediate F is deprotected from Boc under the action of hydrogen chloride, and then the free compound of formula I is obtained by reacting with sodium hydroxide solution. 3 (CH2) m NH2 is reacted with phenyl chloroformate to obtain intermediate E; intermediate D is reacted with intermediate E to obtain intermediate F by condensation reaction, and the hydrochloride salt of the compound of formula I is obtained after intermediate F is deprotected from Boc under the action of hydrogen chloride, and then the free compound of formula I is obtained by reacting with sodium hydroxide solution.

[0096] BRIEF DESCRIPTION OF DRAWINGS

[0097] Figure 1 It is an effect diagram of the anti-inflammatory effect of the compound of the embodiment of the present application, and p<0.05 compared with the solvent group, p<0.01 compared with the solvent group, p<0.01 compared with the celecoxib group;

[0098] Figure 2 It is an effect diagram of the compound of the embodiment of the present application reducing the cerebral infarction volume of cerebral ischemic rats, p<0.05 compared with the solvent group, p<0.05 compared with the edaravone group, and p<0.05 compared with the t-AUCB group;

[0099] Figure 3 It is an effect diagram of the compound of the embodiment of the present application reducing the level of inflammatory factors in the brain tissue of cerebral ischemic rats, p<0.05 compared with the solvent group, p<0.05 compared with the sham operation group, and p<0.05 compared with the t-AUCB group;

[0100] Figure 4 It is an effect diagram of the compound of the embodiment of the present application improving the survival rate of cerebral ischemic rats.

[0101] Figure 5 It is an effect diagram of the compound of the embodiment of the present application improving the learning and memory function of cerebral ischemic rats, p<0.05 compared with the solvent group, p<0.05 compared with the sham operation group, p<0.05 compared with the t-AUCB group, and p<0.05 compared with the edaravone group;

[0102] Figure 6 It is an effect diagram of the hypoglycemic effect of the compound of the embodiment of the present application, p<0.05 compared with the solvent group, p<0.05 compared with the normal group, and p<0.05 compared with the t-AUCB group;

[0103] Figure 7Figure for the anti-fibrosis effect of the compound of the embodiment of the present application, *p<0.05 compared with the solvent group, #p<0.05 compared with the sham operation group, $p<0.05 compared with the t-AUCB group;

[0104] Figure 8 Figure for the analgesic effect of the compound of the embodiment of the present application on selective injury of sciatic nerve branches, *p<0.05 compared with the solvent group, #p<0.05 compared with the EC5026 group;

[0105] Figure 9 Figure for the analgesic effect of the compound of the embodiment of the present application on diabetic pathological nerve pain, *p<0.05 compared with the solvent group, #p<0.05 compared with the pregabalin group;

[0106] Figure 10 Figure for the analgesic effect of the compound of the embodiment of the present application on paclitaxel-induced neuropathic pain, *p<0.05 compared with the solvent group, #p<0.05 compared with the pregabalin group;

[0107] Figure 11 Figure for the effect of the compound of the embodiment of the present application on reducing the level of inflammatory factors in the spinal cord tissue of neuropathic pain animals, *p<0.05 compared with the solvent group, ##p<0.01 compared with the sham operation group, $p<0.05 compared with the EC5026 group;

[0108] Figure 12 Effect of the compound of the embodiment of the present application on the motor coordination function of rats, **p<0.01 compared with before administration;

[0109] Figure 13 Figure for the antidepressant effect of the compound of the embodiment of the present application, *p<0.05 compared with the solvent group, #p<0.05 compared with the normal group;

[0110] Figure 14 Figure for the effect of the compound of the embodiment of the present application on treating dry eye, **p<0.01 compared with the solvent group, ##p<0.01 compared with the normal group, $p<0.05 compared with the cyclosporine eye drop group;

[0111] Figure 15 Figure for the effect of the compound of the embodiment of the present application on improving the level of 14, 15-EET and 14, 15-DHET in the corneal tissue of dry eye mice, *p<0.05 compared with the solvent group, #p<0.05 compared with the normal group, $p<0.05 compared with the cyclosporine eye drop group;

[0112] Figure 16Effect diagram of the compound of the embodiment of the present application in reducing the level of inflammatory factors in the corneal tissue of dry eye mice, *p<0.05 compared with the solvent group, #p<0.05 compared with the normal group, $p<0.05 compared with the cyclosporine eye drop group;

[0113] Figure 17 Effect diagram of the compound of the embodiment of the present application in reducing the retinal vascular permeability of diabetic mice, *p<0.05 compared with the solvent group, #p<0.05 compared with the normal group, $p<0.05 compared with the t-AUCB group;

[0114] Figure 18 Effect diagram of the compound of the embodiment of the present application in protecting the retinal vascular homeostasis of diabetic mice, *p<0.05 compared with the solvent group, #p<0.05 compared with the normal group, $p<0.05 compared with the t-AUCB group; ↑ represents free pericyte, and ▲ represents acellular capillary;

[0115] Figure 19 Distribution results of the compound of the embodiment of the present application in the eye tissue of a Dutch rabbit;

[0116] Figure 20 Effect diagram of the compound of the embodiment of the present application on the content of 14, 15-EET and 14, 15-DHET in rats, *p<0.05 compared with the EC5026 group;

[0117] Figure 21 Molecular docking result diagram of EC5026 and sEH;

[0118] Figure 22 Molecular docking result diagram of the compound of the embodiment 1 of the present application and sEH;

[0119] Figure 23 Molecular docking result diagram of the compound of the embodiment 3 of the present application and sEH;

[0120] Figure 24 Molecular docking result diagram of the compound of the embodiment 16 of the present application and sEH. DETAILED DESCRIPTION

[0121] In the following examples, methods for preparing some of the compounds described in the Preparation section are depicted. It will be appreciated that the following methods, as well as other methods known to those of ordinary skill in the art, can be applied to the preparation of all of the compounds described in the present application. The examples are intended to illustrate and not to limit the scope of the present application.

[0122] The series of 2-substituted aminobenzothiazole compounds related to the present application are tested for sEH inhibitory activity in vitro, and the results show that they all have significant inhibitory activity. In vivo pharmacodynamic tests show that they have good efficacy in inflammation, stroke, diabetes, fibrosis, pain, depression, and dry eye.

[0123] The proton NMR spectra of the compounds were determined using a Bruker ARX-400 or Bruker ARX-600, and the mass spectra were determined using an Agilent 1100LC / MSD. All reagents used were of analytical or chemical purity.

[0124] Example 1 Preparation of 1-{N-[2-(4-morpholinyl)ethyl]-2-aminobenzo[d]thiazolyl-5-yl}-3-(4-chlorophenyl)urea

[0125] 1.1 Preparation of N-tert-butoxycarbonyl-2-amino-5-nitrobenzo[d]thiazole (B)

[0126] At room temperature, 25.5 g (130.6 mmol) of 2-amino-5-nitrobenzo[d]thiazole (intermediate A, commercially available) and 16.0 g (130.6 mmol) of 4-dimethylaminopyridine (DMAP) were added sequentially to 255 mL of dichloromethane, and the mixture was heated to 40 °C. 56.9 g (261.2 mmol) of (Boc)₂O was slowly added dropwise to the reaction mixture, and the reaction was allowed to proceed for 2 h after the addition was complete. After the reaction was complete, the reaction mixture was cooled to room temperature, evaporated to dryness under reduced pressure, and the residue was added to 255 mL of 10% citric acid aqueous solution. The mixture was stirred for 1 h, filtered, and the filter cake was washed with water and dried to give 31.0 g of a pale yellow solid, with a yield of 80.4%.

[0127] ESI-MS m / z: 296.1 [M+H] + .

[0128] Preparation of 1,2N-[2-(4-morpholinyl)ethyl]-N-tert-butoxycarbonyl-2-amino-5-nitrobenzo[d]thiazole (C)

[0129] At room temperature, 5.0 g (16.9 mmol) of intermediate B, 3.8 g (25.4 mmol) of N-(2-chloroethyl)morpholine, and 7.0 g (50.7 mmol) of anhydrous potassium carbonate were added sequentially to 50 mL of dioxane. After the addition was complete, the reaction solution was heated to 90 °C and reacted for 8 h. After the reaction was complete, the mixture was filtered while hot, and the filtrate was evaporated to dryness under reduced pressure to obtain a brown oily liquid. The residue was dissolved in 200 mL of ethyl acetate, washed with water, and evaporated to dryness under reduced pressure to obtain 4.2 g of a pale yellow solid, with a yield of 61.6%.

[0130] ESI-MS m / z: 443.1 [M+Na] + .

[0131] 1.3N 2 -[2-(4-morpholinyl)ethyl]-N 2 Preparation of -tert-butoxycarbonyl-2,5-diaminobenzo[d]thiazole (D)

[0132] At room temperature, 2.3 g (41.1 mmol) of iron powder and 4.4 g (82.4 mmol) of ammonium chloride were added into 45 mL of 95% ethanol, and refluxed for 10 min. 4.2 g (10.3 mmol) of intermediate C was added into the ethanol solution in batches, and after the addition, the reaction was refluxed for 2 h. After the reaction was completed, it was filtered under suction while hot, and the filtrate was evaporated under reduced pressure to obtain 2.5 g of brown solid, with a yield of 65.2%.

[0133] ESI-MS m / z: 379.1 [M+H] + .

[0134] 1.4 Synthesis of phenyl N-(4-chlorophenyl)carbamate (E)

[0135] Under an ice water bath, 1.4 g (11.2 mmol) of 4-chloroaniline and 0.7 g (6.7 mmol) of sodium carbonate were added into a tetrahydrofuran solution, and stirred. Then, 1.9 g (11.9 mmol) of phenyl chloroformate in 3 mL of tetrahydrofuran was added dropwise into the reaction solution, and after the dropwise addition was completed, the reaction was allowed to react at room temperature for 2 h. After the reaction was completed, the reaction solution was evaporated under reduced pressure, 10 mL of water was added into the residue, and stirred. A solid was precipitated, which was filtered under suction, and the filter cake was washed with water and dried to obtain 2.5 g of a solid, with a yield of 91%.

[0136] 1.5 Preparation of 1-{N-[2-(4-morpholinyl)ethyl]-N-tert-butoxycarbonyl-2- aminobenzo[d]thiazol-5-yl}-3-(4-chlorophenyl)urea (F)

[0137] At room temperature, 2.0 g (5.3 mmol) of intermediate D and 0.8 mL (5.8 mmol) of triethylamine were added into 20 mL of dry dioxane, and stirred to dissolve. 1.6 g (6.3 mmol) of intermediate E was added into the reaction solution in batches, and after the addition was completed, the reaction solution was heated to 55°C and reacted for 5 h. After the reaction was completed, the reaction solution was cooled to room temperature, 20 mL of water was added, and stirred. Then, it was filtered under suction and dried to obtain 2.2 g of a white solid, with a yield of 78.6%.

[0138] 1.6 Preparation of 1-{N-[2-(4-morpholinyl)ethyl]-2-aminobenzo[d]thiazol-5-yl}-3-(4- chlorophenyl)urea (Example 1)

[0139] To a solution of 0.19 g (0.36 mmol) of intermediate F in 2 mL of saturated hydrogen chloride in dioxane was added at room temperature and the reaction was heated to 60 °C for 3 h. Upon completion, the reaction was cooled to room temperature and filtered to give the hydrochloride salt of the compound of Example 1. The hydrochloride salt of the compound of Example 1 was added to water and the pH was adjusted to 10-11 with 20% aqueous sodium hydroxide solution. A solid precipitated and was filtered to give 0.12 g of the compound of Example 1 as a white solid in 79.0% yield.

[0140] 1 H NMR (600 MHz, CD3OD) δ 7.76 (s, 1H), 7.50 (d, J = 8.5 Hz, 1H), 7.44 (d, J = 8.8 Hz, 2H), 7.27 (d, J = 8.8 Hz, 2H), 7.09 (dd, J = 8.5, 1.9 Hz, 1H), 3.87 (m, 4H), 3.72 (m, 2H), 3.07 (m, 6H). ESI-MS m / z: 431.9 [M+H] + .

[0141] with appropriate R 4 The key intermediate D was synthesized according to the synthetic method of 1.1-1.3 in Example 1, starting from 2-amino-5-(6-)nitrobenzo[d]thiazole (A); the intermediate E was synthesized according to the synthetic method of 1.4 in Example 1, starting from the aniline with R 3 The intermediate E was synthesized according to the synthetic method of 1.4 in Example 1, starting from the aniline with R

[0142] Preparation of 1-{N-[2-(2-oxo-1-pyrrolidinyl)ethyl]-2-aminobenzo[d]thiazol-5-yl}-3-(4- chlorophenyl)urea

[0143] The key intermediate B was synthesized according to the synthetic method of 1.1 in Example 1, starting from 2-amino-5-nitrobenzo[d]thiazole; the intermediate D was synthesized according to the synthetic method of 1.2-1.3 in Example 1, starting from intermediate B and N-(2-chloroethyl)pyrrolidin-2-one; the intermediate E was synthesized according to the synthetic method of 1.4 in Example 1, starting from 4-chloroaniline; the compound of Example 2 was synthesized according to the synthetic method of 1.5-1.6 in Example 1, starting from intermediate D and intermediate E.

[0144] 1H NMR (400 MHz, CD3OD) δ 7.62 (d, J = 2.0 Hz, 1H), 7.47 (d, J = 8.5 Hz, 1H), 7.43 (d, J = 8.9 Hz, 2H), 7.27 (d, J = 8.9 Hz, 2H), 7.14 (dd, J = 8.5, 2.1 Hz, 1H), 3.64 (m, 2H), 3.57-3.53 (m, 4H), 2.32 (t, J = 8.1 Hz, 2H), 2.0 (m, 2H). MS (ESI) m / z: 428.0 [M-H] - .

[0145] Example 3 Preparation of 1-{N-[2-(4-morpholinyl)ethyl]-2-aminobenzo[d]thiazol-6-yl}-3-(4- chlorophenyl)urea

[0146] Key intermediate D was synthesized according to the synthetic methods of 1.1-1.3 in Example 1, using 2-amino-6-nitrobenzo[d]thiazole as the starting material. Intermediate E was synthesized according to the synthetic method of 1.4 in Example 1, using 4-chloroaniline as the starting material. The compound of Example 3 was prepared according to the synthetic methods of 1.5-1.6 in Example 1, using intermediate D and intermediate E.

[0147] 1 H NMR (400 MHz, CD3OD) δ 7.62 (d, J = 2.0 Hz, 1H), 7.47 (d, J = 8.5 Hz, 1H), 7.43 (d, J = 8.9 Hz, 2H), 7.27 (d, J = 8.9 Hz, 2H), 7.14 (dd, J = 8.5, 2.1 Hz, 1H), 3.64 (m, 2H), 3.57-3.53 (m, 4H), 2.32 (t, J = 8.1 Hz, 2H), 2.0 (m, 2H). MS (ESI) m / z: 428.0 [M-H] - .

[0148] Example 4 Preparation of 1-{N-[2-(dimethylamino)ethyl]-2-aminobenzo[d]thiazol-6-yl}-3-(4- chlorophenyl)urea

[0149] Key intermediate B was synthesized according to the synthetic method of 1.1 in Example 1, using 2-amino-6-nitrobenzo[d]thiazole as the starting material. Intermediate D was synthesized according to the synthetic methods of 1.2-1.3 in Example 1, using intermediate B and N,N-dimethyl-2-chloroethanamine. Intermediate E was synthesized according to the synthetic method of 1.4 in Example 1, using 4-chloroaniline as the starting material. The compound of Example 4 was prepared according to the synthetic methods of 1.5-1.6 in Example 1, using intermediate D and intermediate E.

[0150] MS (ESI) m / z: 390.0 [M+H] + .

[0151] Preparation of 1-{N-[2-(4-morpholinyl)ethyl]-2-aminobenzo[d]thiazol-5-yl}-3-(3- fluoro-4-chlorophenyl)urea

[0152] The key intermediate B was synthesized from 2-amino-5-nitrobenzo[d]thiazole according to the synthetic method of 1.1 in Example 1; the intermediate D was synthesized from the intermediate B and N-(2-chloroethyl)piperidine according to the synthetic method of 1.2-1.3 in Example 1; the intermediate E was synthesized from 3-fluoro-4-chloroaniline according to the synthetic method of 1.4 in Example 1; the compound of Example 5 was prepared from the intermediate D and the intermediate E according to the synthetic method of 1.5-1.6 in Example 1.

[0153] 1 H NMR (400 MHz, CD3OD) δ 7.61-7.58 (m, 2H), 7.48 (d, J = 8.5 Hz, 1H), 7.33 (t, J = 8.5 Hz, 1H), 7.14 (dd, J = 8.5, 2.1 Hz, 1H), 7.09 (m, 1H), 3.71 (t, J = 4.6 Hz, 4H), 3.58 (t, J = 6.5 Hz, 2H), 2.66 (t, J = 6.5 Hz, 2H), 2.54 (t, J = 4.5 Hz, 4H). ESI-MS m / z: 450.5 [M+H] + .

[0154] Preparation of 1-{N-[2-(1-piperidinyl)ethyl]-2-aminobenzo[d]thiazol-5-yl}-3-(4- chlorophenyl)urea

[0155] The key intermediate D was synthesized from 2-amino-5-nitrobenzo[d]thiazole according to the synthetic method of 1.1-1.3 in Example 1; the intermediate E was synthesized from 3-fluoro-4-chloroaniline according to the synthetic method of 1.4 in Example 1; the compound of Example 6 was prepared from the intermediate D and the intermediate E according to the synthetic method of 1.5-1.6 in Example 1.

[0156] 1H NMR (400 MHz, CD3OD) δ 7.65 (s, 1H), 7.49 (d, J = 8.5 Hz, 1H), 7.43 (d, J = 8.8 Hz, 2H), 7.27 (d, J = 8.8 Hz, 2H), 7.12 (dd, J = 8.5, 2.0 Hz, 1H), 3.63 (t, J = 6.4 Hz, 2H), 2.79 (m, 2H), 2.69 (m, 4H), 1.70 (m, 4H), 1.54 (m, 2H). ESI-MS m / z: 430.0 [M+H] + .

[0157] Preparation of 1-{N-[2-(4-morpholinyl)ethyl]-2-aminobenzo[d]thiazol-6-yl}-3-(4- trifluoromethoxyphenyl)urea

[0158] Key intermediate D was synthesized according to the synthetic methods of 1.1-1.3 in Example 1, using 2-amino-6-nitrobenzo[d]thiazole as the raw material. Intermediate E was synthesized according to the synthetic method of 1.4 in Example 1, using 4- (trifluoromethoxy)aniline as the raw material. The compound of Example 7 was prepared according to the synthetic methods of 1.5-1.6 in Example 1, using intermediate D and intermediate E.

[0159] ESI-MS m / z: 481.9 [M+H] + .

[0160] Preparation of 1-{N-[2-(2-oxo-1-pyrrolidinyl)ethyl]-2-aminobenzo[d]thiazol-5-yl}-3-(3- fluoro-4-chlorophenyl)urea

[0161] Key intermediate B was synthesized according to the synthetic method of 1.1 in Example 1, using 2-amino-5-nitrobenzo[d]thiazole as the raw material. Intermediate D was synthesized according to the synthetic methods of 1.2-1.3 in Example 1, using intermediate B and N-(2- chloroethyl)pyrrolidin-2-one. Intermediate E was synthesized according to the synthetic method of 1.4 in Example 1, using 3-fluoro-4-chloroaniline as the raw material. The compound of Example 8 was prepared according to the synthetic methods of 1.5-1.6 in Example 1, using intermediate D and intermediate E.

[0162] 1H NMR (600 MHz, DMSO-d6) δ 9.11 (s, 1H), 8.86 (s, 1H), 8.15 (s, 1H), 7.69 (d, J = 11.7 Hz, 1H), 7.62 (s, 1H), 7.55 (d, J = 8.0 Hz, 1H), 7.47 (t, J = 8.4 Hz, 1H), 7.19 (d, J = 8.4 Hz, 1H), 7.09 (d, J = 8.1 Hz, 1H), 3.51 (s, 4H), 3.43 (m, 4H), 2.18 (t, J = 7.3 Hz, 2H), 1.90 (t, J = 7.0 Hz, 2H). ESI-MS m / z: 448.1 [M+H] + .

[0163] Preparation of 1-{N-[2-(4-morpholinyl)ethyl]-2-aminobenzo[d]thiazol-6-yl}-3-(1- adamantyl)urea

[0164] Key intermediate D was synthesized according to the synthetic method of 1.1-1.3 in Example 1, using 2-amino-6-nitrobenzo[d]thiazole as the raw material. Intermediate E was synthesized according to the synthetic method of 1.4 in Example 1, using adamantylamine as the raw material. The compound of Example 9 was prepared according to the synthetic method of 1.5-1.6 in Example 1, using intermediate D and intermediate E.

[0165] ESI-MS m / z: 456.0 [M+H] + , 477.9 [M+Na] + .

[0166] Preparation of 1-{N-[2-(2-oxo-1-pyrrolidinyl)ethyl]-2-aminobenzo[d]thiazol-6-yl}-3-(4- chlorophenyl)urea

[0167] Key intermediate B was synthesized according to the synthetic method of 1.1 in Example 1, using 2-amino-6-nitrobenzo[d]thiazole as the raw material. Intermediate D was synthesized according to the synthetic method of 1.2-1.3 in Example 1, using intermediate B and N-(2-chloroethyl)pyrrolidin-2-one. Intermediate E was synthesized according to the synthetic method of 1.4 in Example 1, using 4-chloroaniline as the raw material. The compound of Example 10 was prepared according to the synthetic method of 1.5-1.6 in Example 1, using intermediate D and intermediate E.

[0168] 1H NMR (600 MHz, DMSO-d6) δ 10.33 (br s, 1H), 9.54 (s, 1H), 9.51 (s, 1H), 8.10 (d, J = 1.7 Hz, 1H), 7.52 (d, J = 8.7 Hz, 1H), 7.49 (d, J = 8.8 Hz, 2H), 7.43 (dd, J = 8.7, 1.9 Hz, 1H), 7.33 (d, J = 8.8 Hz, 2H), 3.71 (m, 2H), 3.48 (t, J = 5.9 Hz, 2H), 3.44 (t, J = 7.0 Hz, 2H), 2.19 (t, J = 8.0 Hz, 2H), 1.92 (m, 2H). ESI-MS m / z: 429.9 [M+H] + .

[0169] Example 11 Preparation of 1-{N-[2-(3-oxo-4-morpholinyl)ethyl]-2- aminobenzo[d]thiazol-5-yl}-3-(3,4-dichlorophenyl)urea

[0170] The key intermediate B was synthesized according to the synthetic method of 1.1 in Example 1, starting from 2-amino-5-nitrobenzo[d]thiazole; the intermediate B and N-(2- chloroethyl)morpholine-3-one were used to synthesize the intermediate D according to the synthetic method of 1.2-1.3 in Example 1; the intermediate E was synthesized according to the synthetic method of 1.4 in Example 1, starting from 3,4-dichloroaniline; the intermediate D and the intermediate E were used to synthesize the compound of Example 11 according to the synthetic method of 1.5-1.6 in Example 1.

[0171] 1 H NMR (600 MHz, DMSO-d6) δ 9.11 (s, 1H), 8.92 (s, 1H), 8.31 (br s, 1H), 7.89 (d, J = 2.0 Hz, 1H), 7.63 (s, 1H), 7.56 (d, J = 8.4 Hz, 1H), 7.52 (d, J = 8.8 Hz, 1H), 7.34 (dd, J = 8.8, 2.0 Hz, 1H), 7.10 (d, J = 8.4 Hz, 1H), 4.00 (s, 2H), 3.79 (t, J = 4.9 Hz, 2H), 3.56 (m, 4H), 3.42 (t, J = 4.8 Hz, 2H). ESI-MS m / z: 502.4 [M+Na] + .

[0172] Example 12 Preparation of 1-{N-[2-(3-oxo-4-morpholinyl)ethyl]-2-amino-5- fluorobenzo[d]thiazol-6-yl}-3-(4-chlorophenyl)urea

[0173] The key intermediate B was synthesized according to the synthetic method of 1.1 in the example 1, using 2-amino-5-fluoro-6-nitrobenzo[d]thiazole as the raw material; the intermediate D was synthesized according to the synthetic method of 1.2-1.3 in the example 1, using the intermediate B and N-(2-chloroethyl)pyrrolidin-2-one; the intermediate E was synthesized according to the synthetic method of 1.4 in the example 1, using 4-chloroaniline as the raw material; the compound of example 12 was prepared according to the synthetic method of 1.5-1.6 in the example 1, using the intermediate D and the intermediate E.

[0174] 1 H NMR (400 MHz, DMSO-d6) δ 9.31 (s, 1H), 8.53 (s, 1H), 8.30 (d, J = 7.9 Hz, 1H), 8.21 (s, 1H), 7.53 (d, J = 8.9 Hz, 2H), 7.38 (d, J = 8.8 Hz, 2H), 7.34 (d, J = 12.1 Hz, 1H), 4.05 (s, 2H), 3.84 (t, J = 4.8 Hz, 2H), 3.59 (m, 4H), 3.45 (t, J = 4.8 Hz, 2H). ESI-MS m / z: 464.1 [M+H] + .

[0175] Preparation of 1-{N-[2-(2-oxo-1-piperidyl)ethyl]-2-amino-5-fluorobenzo[d]thiazol-6-yl}-3-(4- chlorophenyl)urea

[0176] The key intermediate B was synthesized according to the synthetic method of 1.1 in the example 1, using 2-amino-5-fluoro-6-nitrobenzo[d]thiazole as the raw material; the intermediate D was synthesized according to the synthetic method of 1.2-1.3 in the example 1, using the intermediate B and N-(2-chloroethyl)pyrrolidin-2-one; the intermediate E was synthesized according to the synthetic method of 1.4 in the example 1, using 4-chloroaniline as the raw material; the compound of example 12 was prepared according to the synthetic method of 1.5-1.6 in the example 1, using the intermediate D and the intermediate E.

[0177] 1 H NMR (400 MHz, CD3OD) δ 8.19 (d, J = 7.7 Hz, 1H), 7.73-7.69 (m, 1H), 7.63-7.60 (m, 1H), 7.44 (d, J = 8.9 Hz, 2H), 7.27 (d, J = 8.9 Hz, 2H), 7.19 (d, J = 11.9 Hz, 1H), 3.65-3.60 (m, 4H), 3.43 (t, J = 5.7 Hz, 2H), 2.30 (t, J = 6.3 Hz, 2H), 1.77-1.70 (m, 4H). ESI-MS m / z: 462.1 [M+H] + .

[0178] Preparation of Example 14, 1-{N-[2-(3-oxo-4-morpholinyl)ethyl]-2- aminobenzo[d]thiazol-6-yl}-3-(1-adamantyl)urea

[0179] The key intermediate B was synthesized according to the synthetic method of 1.1 in Example 1, starting from 2-amino-6-nitrobenzo[d]thiazole; the intermediate D was synthesized according to the synthetic method of 1.2-1.3 in Example 1, starting from the intermediate B and N-(2-chloroethyl)morpholine-3-one; the intermediate E was synthesized according to the synthetic method of 1.4 in Example 1, starting from adamantylamine; the Example 14 compound was prepared according to the synthetic method of 1.5-1.6 in Example 1, starting from the intermediate D and the intermediate E.

[0180] Example 14 compound.

[0181] ESI-MS m / z: 469.9 [M+H] + , 491.9 [M+Na] + .

[0182] Preparation of Example 15, 1-{N-[2-(3-oxo-4-morpholinyl)ethyl]-2-amino-5- chlorobenzo[d]thiazol-6-yl}-3-(4-chlorophenyl)urea

[0183] The key intermediate B was synthesized according to the synthetic method of 1.1 in Example 1, starting from 2-amino-5-chloro-6-nitrobenzo[d]thiazole; the intermediate D was synthesized according to the synthetic method of 1.2-1.3 in Example 1, starting from the intermediate B and N-(2-chloroethyl)morpholine-3-one; the intermediate E was synthesized according to the synthetic method of 1.4 in Example 1, starting from 4-chloroaniline; the Example 15 compound was prepared according to the synthetic method of 1.5-1.6 in Example 1, starting from the intermediate D and the intermediate E.

[0184] 1 H NMR (400 MHz, CD3OD) δ 8.23 (s, 1H), 7.46-7.44 (m, 3H), 7.27 (d, J = 8.9 Hz, 2H), 4.07 (s, 2H), 3.83 (t, J = 4.9 Hz, 2H), 3.68 (m, 4H), 3.50 (t, J = 5.2 Hz, 2H). ESI-MS m / z: 501.8 [M+Na] + .

[0185] Preparation of Example 16, 1-{N-[2-(2-oxo-1-pyrrolidinyl)ethyl]-2-amino-5- fluorobenzo[d]thiazol-6-yl}-3-(4-chlorophenyl)urea

[0186] The key intermediate B was synthesized according to the synthetic method of 1.1 in Example 1, using 2-amino-5-fluoro-6-nitrobenzo[d]thiazole as the raw material; the intermediate D was synthesized according to the synthetic method of 1.2-1.3 in Example 1, using the intermediate B and N-(2-chloroethyl)pyrrolidin-2-one; the intermediate E was synthesized according to the synthetic method of 1.4 in Example 1, using 4-chloroaniline as the raw material; and the compound of Example 16 was prepared according to the synthetic method of 1.5-1.6 in Example 1, using the intermediate D and the intermediate E.

[0187] 1 H NMR (400 MHz, DMF-d7) δ 9.46 (s, 1H), 8.68 (d, J = 2.1 Hz, 1H), 8.60 (d, J = 7.9 Hz, 1H), 8.26 (t, J = 5.7 Hz, 1H), 7.80 (d, J = 8.9 Hz, 2H), 7.55 (d, J = 8.9 Hz, 2H), 7.47 (d, J = 12.2 Hz, 1H), 3.84 (q, J = 5.9 Hz, 2H), 3.72-3.69 (m, 4H), 3.10 (m, 2H), 2.93 (m, 2H). ESI-MS m / z: 447.9 [M+H] + .

[0188] Structural formulas of the compounds of Examples 1-16 in Table 1

[0189]

[0190]

[0191] Biological activity test:

[0192] 1. Test of the inhibitory activity of the compounds of some examples on sEH

[0193] The in vitro inhibitory activity of the 2-substituted aminobenzothiazole compounds of general formula I according to the present application on sEH was tested by fluorescence analysis. The control substance was t-AUCB.

[0194] sEH solution and fluorescent substrate PHOME (2-(3-phenyl-oxirane-2-yl)acetic acid 1-cyano-1-(6-methoxy-2-naphthyl)methyl ester) solution were both prepared in 25 mM Bis-Tris (pH 7.0) buffer. Example compounds and t-AUCB were dissolved in DMSO and diluted into a series of different concentrations. 100 μL enzyme solution and 5 μL test compound solution were added to the wells of a 96-well plate, incubated at 37 °C for 5 min, and then 95 μL substrate solution was added, and incubation was continued at 37 °C for 15 min. Blank wells contained only the corresponding volume of Bis-Tris (pH 7.0) buffer and DMSO, and full activity wells contained only sEH and substrate and DMSO. After incubation, the fluorescence intensity was measured at an excitation wavelength of 330 nm and an emission wavelength of 465 nm, and the inhibition rate of different concentrations of compounds was calculated based on the full activity wells, and the IC50values were calculated by spss software. 50

[0195] IC50values of some example compounds and t-AUCB against sEH 50 The data are shown in Table 2.

[0196] Table 2 Inhibitory activity of some example compounds against sEH

[0197] Compound IC 50 (nM) Example 1 3.0 Example 3 0.4 Example 5 1.9 Example 6 8.4 Example 8 1.7 Example 10 0.8 Example 11 0.3 Example 12 0.3 Example 13 1.7 Example 15 0.3 Example 16 0.2 t-AUCB 8.0

[0198] The preliminary in vitro sEH inhibitory activity test results show that the compounds of general formula I to be protected by the present application have good sEH inhibitory activity, and some compounds are equivalent to or significantly better than the positive control t-AUCB.

[0199] 2. Measurement of anti-inflammatory effect of the compounds of Example 1, Example 3, Example 10, Example 15 and Example 16

[0200] Forty-two male Kunming mice (20-25 g) were randomly divided into seven groups, namely the Example 1 group, the Example 3 group, the Example 10 group, the Example 15 group, the Example 16 group, the celecoxib group and the blank solvent group, with 6 mice in each group. An inflammatory swelling model was prepared by subcutaneous injection of 50 μL of a 1% λ-carrageenan saline solution on the foot bottom, and 1 h before modeling, the mice were injected intraperitoneally with 100 mg / kg of a corresponding compound solution (20 mg / mL, 10% PEG 400 and 10% Tween 80 aqueous solution) or blank solvent. The change in paw thickness of the mice was measured, and the degree of paw swelling of the mice was calculated as Figure 1 indicated.

[0201] The results show that the compounds of Example 1, Example 3, Example 10, Example 15 and Example 16 can all significantly inhibit the carrageenan-induced swelling of the mouse feet, and the effect is better than that of celecoxib. The results show that the compounds of Example 1, Example 3, Example 10, Example 15 and Example 16 can all significantly inhibit the carrageenan-induced swelling of the mouse feet, and the effect is better than that of celecoxib.

[0202] 3. Determination of the anti-stroke effect of the compounds of Example 1, Example 3, Example 10, Example 15 and Example 16 3.1 Therapeutic effect on acute stage of ischemic stroke

[0203] Male SD rats were used to prepare middle cerebral artery occlusion (MCAO) reperfusion model by thread method, and the ischemia was restored by pulling out the thread after 2 h. Immediately after reperfusion, 1 mg / kg or 3 mg / kg of the above-mentioned compound solution (1.5 mg / mL, 10% PEG 400 and 10% Tween 80 in water), t-AUCB solution (1.5 mg / mL, 10% PEG 400 and 10% Tween 80 in water), edaravone injection (commercially available, 20 mL:30 mg) or blank solvent were injected into the tail vein, with 8 rats in each group. After 24 h, the rats were sacrificed under anesthesia with 10% chloral hydrate, and the brain tissue was cut into consecutive equidistant 4-5 coronal sections. After staining with 2% TTC (2,3,5-chlorinated triphenyl tetrazolium) solution, the brain slices were photographed, and the infarct volume was calculated, as shown in Figure 2 .

[0204] The results showed that the compounds of Example 1, Example 3, Example 10, Example 15 and Example 16 could significantly reduce the brain infarct volume of MCAO rats, and the effect was better than that of t-AUCB and edaravone.

[0205] Another 30 rats were randomly divided into 5 groups, namely sham operation group, solvent group, Example 3 compound (3 mg / kg) group, Example 16 compound (3 mg / kg) group and t-AUCB (3 mg / kg) group. The MCAO reperfusion model was established according to the above method, and the drug was administered immediately after reperfusion. After 24 h, the ischemic brain tissue was weighed, and PBS buffer solution was added at a ratio of 1:9 by weight and volume, and the tissue grinder was ground at low temperature for 1 min, and centrifuged at 3000 rpm for 20 min at 4°C, and the supernatant was collected. The TNF-α and IL-1β enzyme-linked immunosorbent assay kit (refer to the ELISA kit instructions) was used to determine the concentration of TNF-α and IL-1β in the brain tissue, as shown in Figure 3 .

[0206] The results showed that the compounds of Example 3 and Example 16 could significantly reduce the inflammatory factor level in the brain tissue of MCAO rats, and the effect was better than that of t-AUCB.

[0207] 3.2 Long-term therapeutic effect on ischemic stroke

[0208] Male SD rats (78) were prepared for MCAO reperfusion model, divided into sham operation group, solvent group, example 3 compound (3 mg / kg) group, example 16 compound (3 mg / kg) group, t-AUCB (3 mg / kg) group and edaravone (3 mg / kg) group. Sham operation group 8, the rest of each group of 14, after ischemia reperfusion, intravenous injection of drugs once a day (experimental drugs same as 3.1) for two weeks. The survival of animals was recorded every day, and the results are shown in Figure 4 The Morris water maze experiment was started in the third week. The rats were placed in the water facing the pool wall, and the latency period for finding the platform within 60s was recorded. If the platform was found within 60s, the rat was guided to the platform and stayed for 30s. The latency period for the rat to find the platform within 4 days was recorded to evaluate the spatial memory ability of the rat, and the experimental results are shown in Figure 5

[0209] The results show that both example 3 and example 16 compounds can improve the survival rate of rats, significantly reduce the escape latency of rats, and enhance the learning and memory ability, which is better than t-AUCB and edaravone.

[0210] 4. Determination of the hypoglycemic effect of example 1, example 3, example 10, example 15 and example 16 compounds

[0211] Male BALB / c mice (18-22g) were randomly divided into 8 groups, namely normal control group, blank solvent group, example 1 group, example 3 group, example 10 group, example 15 group, example 16 group and t-AUCB group, 6 in each group. Except for the normal control group, the mice were intraperitoneally injected with 1% streptozotocin (STZ) solution (0.1 mol / L citric acid buffer solution, pH 4.5) at a dose of 50 mg / kg for 5 consecutive days, once a day, to prepare a diabetic model. On the first day of modeling, the mice were intraperitoneally injected with 10 mg / kg of the corresponding compound solution (6 mg / mL, 10% PEG 400 and 10% Tween 80 aqueous solution) or blank solvent once a day. Two weeks later, the fasting blood glucose of the mice was measured as shown in Figure 6

[0212] The results show that the blood glucose of the mice in the administration group is significantly lower than that in the blank solvent group, and example 1, example 3, example 10, example 15 and example 16 compounds can alleviate STZ-induced hyperglycemia, and example 3 and example 16 compounds are better than t-AUCB.

[0213] 5. Determination of the anti-fibrosis effect of example 1, example 3, example 10, example 15 and example 16 compounds

[0214] ​​Male C57BL6 / J mice (18-22 g) were randomly divided into 8 groups, i.e. sham operation group, blank solvent group, Example 1 group, Example 3 group, Example 10 group, Example 15 group, Example 16 group and t-AUCB group, 6 mice in each group. After the mice were anesthetized, the left ureter was ligated and cut to prepare a unilateral ureteral obstruction (UUO) model. The sham operation group only separated the ureter without ligation. Starting 3 days before modeling, the mice were intraperitoneally injected with 10 mg / kg of the corresponding compound solution (the experimental drug was prepared as in 4) or blank solvent, once a day. After 7 days of modeling, the animals were sacrificed, and the kidneys were taken for Masson staining to detect histopathological changes, and the experimental results are shown in Figure 7 .

[0215] The results show that the compounds of Example 1, Example 3, Example 10, Example 15 and Example 16 can significantly reduce the fibrosis area in the kidney tissue of mice, and the compounds of Example 3 and Example 16 are better than t-AUCB.

[0216] 6. Measurement of analgesic effect of the compounds of Example 1, Example 3, Example 10, Example 15 and Example 16

[0217] (1) The initial mechanical withdrawal threshold (MWT) of the foot sole of male SD rats (200-250 g) was measured by Von Frey electronic pain tester, and then the rats were anesthetized, the skin was longitudinally incised on the left femur, the muscles were bluntly separated, the sciatic nerve trunk and its distal branches were exposed, the surrounding adhesion tissues and 3 branches of the sciatic nerve, i.e. tibial nerve, common peroneal nerve and sural nerve were separated, the tibial nerve and common peroneal nerve were tightly ligated with silk thread, and about 2-4 mm of the nerve was cut off at the distal end of the ligation, the sural nerve was reserved and avoided to be injured, to prepare a sciatic nerve branch selective injury (SNI) pain model. After one week of operation, the MWT was measured to confirm the SNI model.

[0218] After the model was successfully established, the SNI model rats were divided into 8 groups, i.e. Example 1 group, Example 3 group, Example 10 group, Example 15 group, Example 16 group, EC5026 group, pregabalin group and blank solvent group, 6 rats in each group, and the corresponding compound solution (1.2 mg / mL, 10% PEG 400 and 10% Tween 80 aqueous solution) or blank solvent was administered by gavage, and the dose was 3 mg / kg. Taking the MWT before administration as 100%, the change of MWT at different time points after administration was measured, the MWT% -time curve was drawn, and the area under the curve (AUC) was calculated. The experimental results are shown in Figure 8 .

[0219] (2) Male SD rats (180-220 g) were injected intraperitoneally with 1% streptozotocin (STZ) solution (0.1 mol / L citric acid buffer with pH value of 4.5) at a dose of 60 mg / kg to prepare a diabetic model. One week later, the rats were confirmed to have a diabetic model when the fasting blood glucose was greater than 11.1 mmol / L. Two weeks later, the rats were confirmed to have a diabetic neuropathic pain (DNP) model when the MWT was measured.

[0220] The DNP model rats were divided into 7 groups, i.e. Example 1 group, Example 3 group, Example 10 group, Example 15 group, Example 16 group, pregabalin group and blank solvent group, each group of 6 rats, and each group was given a corresponding compound solution (1.2 mg / mL, 10% PEG 400 and 10% Tween 80 aqueous solution) or blank solvent by gavage at a dose of 3 mg / kg. The MWT at different time points after administration was measured with the MWT before administration as 100%, and the MWT% -time curve was plotted to calculate the area under the curve (AUC). The experimental results are shown in Table 2. Figure 9

[0221] (3) C57BL6 / J mice (18-22 g) were injected intraperitoneally with paclitaxel solution (0.2 mg / mL, solvent: DMSO-PEG400-Tween80-normal saline (1:1:1:7, V / V)) at a dose of 2 mg / kg at 1d, 3d, 5d and 7d. The MWT of the mice was monitored every 4 days after the start of modeling using a Von Frey fiber pain tester to confirm the paclitaxel-induced neuropathic pain (PINP) model.

[0222] After the model was successfully established, the PINP model mice were divided into 7 groups, i.e. Example 1 group, Example 3 group, Example 10 group, Example 15 group, Example 16 group, pregabalin group and blank solvent group, each group of 6 rats, and each group was given a corresponding compound solution (3.75 mg / mL, 10% PEG 400 and 10% Tween 80 aqueous solution) or blank solvent by gavage at a dose of 30 mg / kg. The MWT at different time points after administration was measured with the MWT before administration as 100%, and the MWT% -time curve was plotted to calculate the area under the curve (AUC). The experimental results are shown in Table 4. Figure 10

[0223] The results show that the compounds of Example 1, Example 3, Example 10, Example 15 and Example 16 can significantly improve the MWT of SNI rats, DNP rats and PINP mice, and relieve neuropathic pain. The effects of Example 3 and Example 16 compounds are better than those of EC5026 and pregabalin.

[0224] 6.2 Measurement of the level of inflammatory factors in the spinal cord

[0225] ​​A batch of SD rats (200-250 g) 30 were randomly divided into 5 groups, namely, Example 3 group, Example 16 group, EC5026 group, blank solvent group and sham operation group, 6 in each group. According to the method under item 6.1, SNI model was prepared, and the sham operation group only separated the tibial nerve, common peroneal nerve and sural nerve. After the model was successfully established, the corresponding compound solution (1.2 mg / mL, 10% PEG400 and 10% Tween80 aqueous solution) or blank solvent was given by gavage, and the dose was 3 mg / kg. 2h after administration, the animals were anesthetized, and about 200 mL of normal saline and about 100 mL of 4% paraformaldehyde were used for heart perfusion in turn. The L4-L6 segment of the spinal cord was taken, weighed, and then added to PBS buffer solution at a ratio of 1 g:9 mL at 4°C, and homogenized. The tissue homogenate was centrifuged at 4°C, 2500 r / min for 10 min, and the supernatant was used. The concentration of inflammatory factors TNF-α and IL-1β in the rat spinal cord was determined by TNF-α and IL-1β enzyme-linked immunosorbent assay kit (according to the kit instructions). Figure 11

[0226] The results show that the compounds of Example 3 and Example 16 can significantly reduce the level of inflammatory factors in the spinal cord of SNI rats, and the effect is better than that of pregabalin.

[0227] 6.3 Effect on motor coordination function of rats

[0228] The effect of the compounds on the motor coordination function of rats was evaluated by using a rotating rod fatigue instrument. 42 male SD rats were randomly divided into 7 groups, namely, Example 1 group, Example 3 group, Example 10 group, Example 15 group, Example 16 group, pregabalin group and normal control group, 6 in each group. 3 days before the test, the rats were trained 3 times a day at a speed of 10-20 r / min for 5 min each time, and the interval between two training times was 20 min for fatigue recovery. On the test day, the corresponding compound solution (8 mg / mL, 10% PEG400 and 10% Tween80 aqueous solution) or blank solvent was given by gavage, and the dose was 20 mg / kg. The walking time of rats on the rotating rod at a speed of 25 r / min was recorded before and after administration. The walking time of rats on the rotating rod was statistically analyzed as shown in Figure 12

[0229] The results show that the walking time of rats on the rotating rod in the pregabalin group decreased significantly, and the compounds of Example 1, Example 3, Example 10, Example 15 and Example 16 did not affect the walking time of rats on the rotating rod, without adverse reactions of the central nervous system caused by pregabalin.

[0230] 7. Anti-depression effect determination of Example 1, Example 3, Example 10, Example 15 and Example 16 compounds

[0231] ​​Male Kunming mice (20-25 g) were randomly divided into 7 groups, i.e. Example 1 group, Example 3 group, Example 10 group, Example 15 group, Example 16 group, blank solvent group and normal control group, 6 mice in each group. The Kunming mice were first pre-swimming for 10 min, except for the normal control group, the mice were intraperitoneally injected with 0.5 mg / kg of lipopolysaccharide saline solution (0.1 mg / mL), and 23 h later, subcutaneously injected with 10 mg / kg of the corresponding compound solution (6 mg / mL, 10% PEG 400 and 10% Tween 80 aqueous solution) or blank solvent, and 1 h later, forced swimming test was performed for 6 min, and the immobile time (s) of mice in each group within the last 4 min was recorded. The immobile time of mice in forced swimming was statistically analyzed as shown in Table 1. Figure 13

[0232] The results showed that the compounds of Example 1, Example 3, Example 10, Example 15 and Example 16 could significantly reduce the immobile time of mice in forced swimming.

[0233] 8. The determination of the effect of relieving dry eye of the compounds of Example 1, Example 3, Example 10, Example 15 and Example 16

[0234] Male BALB / c mice (18-22 g) were randomly divided into 8 groups, i.e. Example 1 group, Example 3 group, Example 10 group, Example 15 group, Example 16 group, cyclosporine eye drops group, blank solvent group and normal control group, 9 mice in each group. Except for the normal control group, the mice were treated with 0.2% benzalkonium chloride (BAC) saline solution eye drops, 3 times a day, for 15 consecutive days, to establish a dry eye model. The above-mentioned compounds (2 mg / mL, 10% hydroxypropyl-β-cyclodextrin solution), cyclosporine eye drops (II) (commercially available, 0.4 mL:0.2 mg) or blank solvent were used for intervention treatment, 3 times a day, 5 μL per eye for each mouse, for 15 consecutive days. The tear secretion of mice and corneal fluorescein sodium staining scores were measured at different times, as shown in Table 2. Figure 14

[0235] The results showed that the compounds of Example 1, Example 3, Example 10, Example 15 and Example 16 could significantly relieve BAC-induced dry eye, improve the tear secretion of mice, and relieve corneal damage, and the effect was better than that of cyclosporine eye drops (II).

[0236] ​​Another 75 male BALB / c mice (18-22 g) were randomly divided into 5 groups, namely, Example 3 group, Example 16 group, cyclosporine eye drops (II) group, blank solvent group and normal control group, 15 mice in each group. The dry eye model and administration were prepared according to the above method, and 15 days later, the mice were sacrificed to take corneas, 6 corneas of every 3 mice were combined as one sample, 3 samples in each group were used to determine the levels of 14, 15-EET and 14, 15-DHET in the tissue. The remaining corneas were combined as one sample for every 4 mice, 3 samples in each group were used to determine the concentration of TNF-α in the tissue. The levels of 14, 15-EET and 14, 15-DHET were determined by LC-MS / MS method, and the concentration of TNF-α was determined by enzyme-linked immunoassay kit according to the kit instructions. The levels of 14, 15-EET and 14, 15-DHET in the tissue are shown in Figure 15 The concentration of inflammatory factor TNF-α in the tissue is shown in Figure 16

[0237] The results show that the compounds of Example 3 and Example 16 can increase the level of 14, 15-EET in the cornea, and reduce the contents of 14, 15-DHET and TNF-α, and the effect is better than that of cyclosporine eye drops (II).

[0238] 9. Therapeutic effect of Example 3 and Example 16 compounds on diabetic retinopathy

[0239] 80 male C57BL / 6J mice (18-22 g) were randomly divided into 5 groups, namely, Example 3 group, Example 16 group, t-AUCB group, blank solvent group and normal control group, 16 mice in each group. Except for the normal control group, the mice were intraperitoneally injected with 1% streptozotocin (STZ) solution (0.1 mol / L citric acid buffer solution, pH 4.5), the dose was 50 mg / kg, once a day, for 5 consecutive days. One week later, the mice with random blood glucose ≥16.7 mmol / L were considered to have successfully established a diabetic model, and the eye drops of Example 3, Example 16 compound (1 mg / mL, 10% hydroxypropyl-β-cyclodextrin solution) or blank solvent were used for intervention treatment, twice a day, 5 μL per eye per time. The diabetic mice were fed for 6 months to induce retinopathy, and 6 months later, the mice were injected with 2% Evans blue saline solution 0.1 mL in the tail vein, circulated for 2 h, and the retinal tissue was taken after 50 mL of physiological saline was perfused into the heart. After homogenization and centrifugation, the supernatant was taken, and the absorbance was measured at 620 nm to investigate the retinal vascular permeability. Another retinal tissue was treated with 5% pepsin solution and pepsin, and the excess tissue was blown off to obtain retinal blood vessels. After PAS staining, the retinal blood vessels and pericyte morphology were observed under a microscope, and the number of free pericytes in the retina was counted. The experimental results are shown in Figure 17 Figure 18 .​​

[0240] The results show that the compounds of Example 3 and Example 16 can significantly reduce the permeability of retinal blood vessels of diabetic mice, reduce the number of free pericytes and acellular vessels in retinal capillaries, maintain vascular homeostasis, and alleviate the symptoms of diabetic retinopathy, and the effect is better than that of t-AUCB.

[0241] The distribution of the compounds of Example 3 and Example 16 in eye tissues was studied using Dutch rabbits. Six male Dutch rabbits (1.6-2.4 kg) were divided into two groups of three, and were respectively given eye drops of the compounds of Example 3 and Example 16, 50 μL per eye. The animals were sacrificed 0.5 h after administration, and cornea, aqueous humor, bulbar conjunctiva, iris, vitreous body, choroid, retina and sclera tissue samples were immediately collected, added with 20% methanol at a weight-volume ratio of 1:10, homogenized, and the compound content in the supernatant was determined by LC-MS / MS. The experimental results are shown in Table 2. Figure 19

[0242] The results show that the compounds of Example 3 and Example 16 can reach the retina tissue in the posterior part of the eye after eye administration.

[0243] 10. Effect of the compounds of Example 3 and Example 16 on the content of 14, 15-EET and 14, 15-DHET in rats in vivo

[0244] Male SD rats (250-300 g) were given the compounds of Example 3 and Example 16 and EC5026 solution (8 mg / mL, 10% PEG 400 and 10% Tween 80 aqueous solution) by gavage at a dose of 20 mg / kg, with 6 rats in each group. About 0.25 mL of blood was collected from the jugular sinus before administration, 15 min, 30 min, 1 h, 2 h, 4 h and 8 h after administration, and placed in an EDTA-K2 anticoagulant tube. The whole blood sample was centrifuged at 1500-1600 rpm for 10 min, and the separated plasma was stored in a refrigerator at -40 to -20°C. The concentrations of 14, 15-EET and 14, 15-DHET in rat plasma were determined by liquid chromatography-mass spectrometry. The results are shown in Table 3. Figure 20

[0245] The results show that the compounds of Example 3, Example 16 and EC5026 can significantly increase the level of 14, 15-EET and reduce the level of 14, 15-DHET in rat plasma, and the effect is better than that of EC5026.

[0246] 11. Molecular docking results of EC5026, the compounds of Example 1, Example 3 and Example 16 with sEH

[0247] The sEH co-crystal protein (PDB code: 4OCZ) was used as the docking model, and the compounds of Example 1, Example 3 and Example 16 were docked into the active site of sEH using the software​​ The Glide 4.5.208 software was used to perform molecular docking of EC5026, Example 1, Example 3, and Example 16 compounds with sEH, respectively.

[0248] The results of the molecular docking of EC5026 with sEH are shown in FIG. 1, wherein the 3-fluoro-4-(trifluoromethoxy)phenyl group in the structure of EC5026 is oriented toward the hydrophobic region inside the sEH protein cavity, the phenyl group forms a π-π stacking interaction with the His524 residue, and the 3-fluorine atom forms a halogen bond with the Phe267 residue; the two NHs of the urea fragment form bidentate hydrogen bond interactions with the Asp335 residue, and the carbonyl oxygen atoms form hydrogen bond interactions with the Tyr383 and Tyr466 residues, respectively; the (S)-2-methylbutyramide fragment is oriented toward the solvent region. Figure 21 The results of the molecular docking of Example 1 with sEH are shown in FIG. 2, wherein the 4-chlorophenyl group in the structure of Example 1 is oriented toward the hydrophobic region inside the sEH protein cavity and forms a π-π stacking interaction with the Phe267 residue; the benzothiazole structure forms an additional π-π stacking interaction with the Trp336 residue, and the nitrogen atom in the thiazole ring forms a hydrogen bond interaction with Gln384; the two NHs of the urea fragment form bidentate hydrogen bond interactions with the Asp335 residue, and the oxygen atoms of the urea fragment form hydrogen bond interactions with the Tyr383 and Tyr466 residues, respectively; the (4-morpholinyl)ethyl moiety is oriented toward the solvent region.

[0249] Figure 22 The results of the molecular docking of Example 3 with sEH are shown in FIG. 3, wherein the 4-chlorophenyl group in the structure of Example 3 is oriented toward the hydrophobic region inside the sEH protein cavity and forms a π-π stacking interaction with the Phe267 residue; the benzothiazole structure forms an additional π-π stacking interaction with the Trp336 residue; the two NHs of the urea fragment form bidentate hydrogen bond interactions with the Asp335 residue, and the oxygen atoms of the urea fragment form hydrogen bond interactions with the Tyr383 and Tyr466 residues, respectively; the (4-morpholinyl)ethyl moiety is oriented toward the solvent region.

[0250] The results of the molecular docking of Example 16 with sEH are shown in FIG. 4, wherein the 4-chlorophenyl group in the structure of Example 16 is oriented toward the hydrophobic region inside the sEH protein cavity and forms a π-π stacking interaction with the Phe267 residue; the benzothiazole structure forms an additional π-π stacking interaction with the Trp336 residue, and the F atom on the benzene ring forms an additional halogen bond interaction with the Asp335 residue; the two NHs of the urea fragment form bidentate hydrogen bond interactions with the Asp335 residue, and the oxygen atoms of the urea fragment form hydrogen bond interactions with the Tyr383 and Tyr466 residues, respectively; the (2-oxo-l-pyrrolidinyl)ethyl moiety is oriented toward the solvent region, and the ketone carbonyl forms an additional hydrogen bond interaction with the Ile363 residue. Figure 23

[0251] The results of the molecular docking of Example 16 with sEH are shown in FIG. 4, wherein the 4-chlorophenyl group in the structure of Example 16 is oriented toward the hydrophobic region inside the sEH protein cavity and forms a π-π stacking interaction with the Phe267 residue; the benzothiazole structure forms an additional π-π stacking interaction with the Trp336 residue, and the F atom on the benzene ring forms an additional halogen bond interaction with the Asp335 residue; the two NHs of the urea fragment form bidentate hydrogen bond interactions with the Asp335 residue, and the oxygen atoms of the urea fragment form hydrogen bond interactions with the Tyr383 and Tyr466 residues, respectively; the (2-oxo-l-pyrrolidinyl)ethyl moiety is oriented toward the solvent region, and the ketone carbonyl forms an additional hydrogen bond interaction with the Ile363 residue. Figure 24 ​​​

[0252] In comparison with EC5026, the compounds of Example 1, Example 3 and Example 16 not only maintain the key interaction force with sEH protein equivalent to EC5026, but also produce additional interaction force with sEH from the benzothiazole structure, which enables better binding with sEH protein, thus exhibiting better inhibitory activity on sEH.

[0253] In summary, in the compounds of general formula I of the present application, the structural modification of substituents R 1 , R 2 , R 3 , R 4 , m, n, X, Y, etc. enables better binding with sEH, thus resulting in good inhibitory effect on sEH and good therapeutic effect on inflammation, stroke, diabetes, fibrosis, pain, depression and dry eye. It can be seen that the interaction of specific substituents with sEH will achieve unexpected effects.

Claims

1. A 2-substituted aminobenzothiazole compound, characterized by: ###0001### 2-Substituted amino benzothiazole compounds are compounds as shown in general formula I and pharmaceutically acceptable salts thereof, wherein, X represents a S atom when Y represents a N atom; X represents a N atom when Y represents a S atom; R 1 and R 2 together with the nitrogen atom to which they are attached form R 3 is phenyl or adamantyl; said phenyl is optionally substituted with 1-3 identical or different R 6 substituents; R 6 R is halogen; R 4 is hydrogen, fluorine or chlorine; m is 0; n is 2.

2. The 2-substituted aminobenzothiazole compound according to claim 1, characterized by: In the compound X represents a S atom when Y represents a N atom; when Y represents a S atom, X represents a N atom, and R 4 is H; R 1 and R 2 with the nitrogen atom to which they are attached R 3 is phenyl, which phenyl is substituted at the para position by R 6 ; R 6 R is halogen; R 4 is hydrogen, fluorine or chlorine; m is 0; n is 2.

3. The 2-substituted aminobenzothiazole compound according to claim 2, wherein: In the compound X represents a S atom when Y represents a N atom, which is shown in general formula (II), R 1 and R 2 with the nitrogen atom to which they are attached form R 3 is phenyl, which phenyl is substituted at the para position by R 6 ; R 6 R is halogen; R 4 is hydrogen, fluorine or chlorine; m is 0; n is 2.

4. The 2-substituted aminobenzothiazole compound according to claim 2, wherein: The compound is 1-{N-[2-(4-morpholinyl)ethyl]-2-amino benzothiazol-5-yl}-3-(4-chlorophenyl) urea; 1-{N-[2-(2-oxo-1-pyrrolidinyl)ethyl]-2-amino benzothiazol-5-yl}-3-(4-chlorophenyl) urea; 1-{N-[2-(4-morpholinyl)ethyl]-2-amino benzothiazol-6-yl}-3-(4-chlorophenyl) urea; 1-{N-[2-(4-morpholinyl)ethyl]-2-amino benzothiazol-5-yl}-3-(3-fluoro-4-chlorophenyl) urea; 1-{N-[2-(1-piperidinyl)ethyl]-2-amino benzothiazol-5-yl}-3-(4-chlorophenyl) urea; 1-{N-[2-(4-morpholinyl)ethyl]-2-amino benzothiazol-6-yl}-3-(4-trifluoromethoxyphenyl) urea; 1-{N-[2-(2-oxo-1-pyrrolidinyl)ethyl]-2-amino benzothiazol-5-yl}-3-(3-fluoro-4-chlorophenyl) urea; 1-{N-[2-(4-morpholinyl)ethyl]-2-amino benzothiazol-6-yl}-3-(1-adamantyl) urea; 1-{N-[2-(2-oxo-1-pyrrolidinyl)ethyl]-2-amino benzothiazol-6-yl}-3-(4-chlorophenyl) urea; 1-{N-[2-(3-oxo-4-morpholinyl)ethyl]-2-amino benzothiazol-5-yl}-3-(3,4-dichlorophenyl) urea; 1-{N-[2-(3-oxo-4-morpholinyl)ethyl]-2-amino-5-fluoro benzothiazol-6-yl}-3-(4-chlorophenyl) urea; 1-{N-[2-(2-oxo-1-piperidinyl)ethyl]-2-amino-5-fluoro benzothiazol-6-yl}-3-(4-chlorophenyl) urea; 1-{N-[2-(3-oxo-4-morpholinyl)ethyl]-2-amino benzothiazol-6-yl}-3-(1-adamantyl) urea; 1-{N-[2-(3-oxo-4-morpholinyl)ethyl]-2-amino-5-chloro benzothiazol-6-yl}-3-(4-chlorophenyl) urea; 1-{N-[2-(2-oxo-1-pyrrolidinyl)ethyl]-2-amino-5-fluoro benzothiazol-6-yl}-3-(4-chlorophenyl) urea.

5. The 2-substituted aminobenzothiazole compound according to any one of claims 1 to 4, characterized in that: The pharmaceutically acceptable salt of the 2-substituted aminobenzothiazole compound is a salt of the 2-substituted aminobenzothiazole compound with an acid selected from hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid, formic acid, acetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, picric acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, benzenesulfonic acid, naphthalenesulfonic acid, trifluoroacetic acid, or aspartic acid.

6. A pharmaceutical composition, characterized by: The pharmaceutical composition contains one or more of the 2-substituted aminobenzothiazole compounds and pharmaceutically acceptable salts thereof according to any one of claims 1-4 and a pharmaceutically acceptable carrier.

7. Use of the 2-substituted aminobenzothiazole compound and pharmaceutically acceptable salts thereof according to any one of claims 1-4 or the pharmaceutical composition according to claim 6 in the preparation of a medicament for treating and / or preventing a sEH-mediated disease.

8. Use according to claim 7, characterized in that: The disease includes an inflammatory disease, a cardiovascular and cerebrovascular disease, diabetes, a diabetic complication, a diabetes-related disease, a fibrotic disease, a neurological and psychiatric disease, pain, an ulcerative disease, and dry eye.

9. Use according to claim 8, characterized in that, The inflammatory disease includes an inflammatory liver disease, an inflammatory kidney disease, an inflammatory lung disease, an inflammatory brain disease, myocarditis, pancreatitis, arthritis, soft tissue inflammation, bone tissue inflammation, and vascular inflammation; the cardiovascular and cerebrovascular disease includes hypertension, myocardial infarction, heart failure, coronary heart disease, cardiovascular arteriosclerosis, ischemic stroke, and hemorrhagic stroke; the diabetes includes type I diabetes or type II diabetes; the diabetic complication includes diabetic retinopathy, diabetes-related uveitis, diabetic cataract, diabetic nephropathy, diabetic skin disease, and diabetic peripheral neuropathy; the diabetes-related disease includes hyperlipidemia, hyperuricemia and gout, obesity, and metabolic syndrome; the fibrotic disease is pulmonary fibrosis, liver fibrosis, myocardial fibrosis, and kidney fibrosis; the neurological and psychiatric disease includes Alzheimer's disease, epilepsy, Parkinson's disease, amyotrophic lateral sclerosis, schizophrenia, mental disorder, depression, and neurasthenia; the pain disease includes neuropathic pain, inflammatory pain, cancer pain, and mixed pain; the ulcerative disease is gastric ulcer, duodenal ulcer, ulcerative colitis, corneal ulcer, and oral ulcer; and the dry eye includes aqueous deficiency dry eye, lipid abnormality dry eye, mucin abnormality dry eye, tear dynamics abnormality dry eye, and mixed dry eye.

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