2-substituted aminobenzothiazole compounds and use thereof

Abstract

The present invention relates to 2-substituted aminobenzothiazole compounds and pharmaceutically acceptable salts thereof, as well as pharmaceutical compositions containing the compounds, and a use thereof. The 2-substituted aminobenzothiazole compounds are compounds represented by general formula I and pharmaceutically acceptable salts thereof, the substituents R1, R2, R3, R4, m, n, X and Y having the meanings provided in the description. The present invention further relates to a use of the compounds represented by general formula I in the preparation of a medicament for treating and / or preventing sEH-mediated diseases, particularly in the preparation of a medicament for treating inflammatory diseases, cardiovascular and cerebrovascular diseases, pain, dry eye syndrome, diabetes, diabetic complications, diabetes-related diseases, fibrotic diseases, neurological and psychiatric diseases, ulcerative diseases, and the like.

Description

2-Substituted aminobenzothiazole compounds and their uses Technical fields:

[0001] This invention relates to the field of medicinal chemistry, and in particular to a series of 2-substituted aminobenzothiazole compounds, their preparation methods, and their use in the preparation of drugs for treating and / or preventing sEH-mediated diseases. Background technology:

[0002] Epoxide hydrolases (EHs) are ubiquitous in nature, found in mammals, plants, and microorganisms. They efficiently catalyze the hydrolysis of epoxides to produce vicinal diols. Eight subtypes of epoxide hydrolases have been identified, including cholesterol 5,6-oxide hydrolase (ChEH), soluble epoxide hydrolase (sEH), and microsomal epoxide hydrolase (mEH). Among these, sEH is one of the most extensively studied epoxide hydrolases in recent years, and it represents a potential therapeutic target for many diseases, including hypertension, lung disease, diabetes, pain, inflammation, and other immune system disorders.

[0003] sEH is encoded by the EPHX2 gene, located in the P21-P12 region of human chromosome 8. This gene comprises 18 introns and 19 exons, encoding over 550 amino acids in a total length of 45 kb. sEH consists of two antiparallel 60 kDa subunits, each containing two distinct enzymatic domains. The C-terminus possesses an α / β hydrolase fold structure, acting as a catalytic subunit to hydrolyze epoxides to form the corresponding diols; the N-terminus exhibits phosphatase activity, capable of hydrolyzing various lipid phosphates. sEH is widely distributed in numerous organs of the human body, including the brain, liver, and kidneys, but its content is lower in organs such as the lungs and spleen. 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 that can be metabolized by cyclooxygenase (COX) and lipoxygenase (LOX) into inflammatory mediators such as prostaglandins, lipoxygenins, and leukotrienes. AA can be converted into more bioactive epoxyeicosatrienoic acids (EETs) by cytochrome P450 enzymes (CYP450). EETs have anti-inflammatory, anti-apoptotic, and antioxidant effects and participate in the pathophysiological processes of various diseases affecting the immune, nervous, and metabolic systems. Sedimentary epoelectrohydrin (sEH) is the main metabolic enzyme of EETs, rapidly hydrolyzing them into less bioactive dihydroxyeicosatrienoic acids (DHETs), thus affecting the protective effects of EETs. sEH inhibitors can maintain the bioactivity of EETs by increasing the level of endogenous EETs, thereby achieving the goal of treating various diseases.

[0005] Currently, no sEH inhibitors are used clinically. Reported sEH inhibitors in the literature include t-AUCB, AR9281, GSK2256294, and EC5026. GSK2256294, a small-molecule sEH inhibitor discovered by GlaxoSmithKline, is intended for the treatment of diseases such as diabetes and obesity and is currently in Phase II clinical trials. EC5026, a urea-based sEH inhibitor discovered by Bruce Hammock's team, is currently in Phase I clinical trials. Summary of the Invention:

[0006] The purpose of this invention is to provide a 2-substituted aminobenzothiazole compound suitable for preparing drugs for treating and / or preventing sEH-mediated diseases.

[0007] To achieve the above objectives, the present invention adopts the following technical solution:

[0008] This invention relates to 2-substituted aminobenzothiazole compounds as shown in general formula I and pharmaceutically acceptable salts thereof.

[0009] in,

[0010] When Y represents N atom, X represents S atom; when Y represents S atom, X represents N atom.

[0011] R 1 and R 2 Each is independently selected from hydrogen, (C1-C6)alkyl, (C3-C7)cycloalkyl; or R 1and R 2 Together with the nitrogen atom to which they are attached, they form a 4-10 membered heterocyclic group, which, in addition to the nitrogen atom, optionally contains 0-3 heteroatoms selected from N, O, or S. The heterocyclic group may also optionally be bonded by 1-3 identical or different R atoms. 5 Substitution: Heterocyclic groups contain 0-2 carbon-carbon double bonds;

[0012] R 5 It can be a hydrogen atom, =O, hydroxyl, amino, halogen, carboxyl, cyano, nitro, (C1-C6)alkyl, (C1-C6)alkoxy, (C3-C7)cycloalkyl, (C1-C6)alkylsulfinyl, (C1-C6)alkylsulfonyl or (C1-C6)alkylformyl.

[0013] R 3 For (C6-C) 10 ) aryl, 5-10-membered heteroaryl or adamantyl, wherein the heteroaryl contains 1-3 heteroatoms selected from N, O or S, and the aryl or heteroaryl may optionally be surrounded by 1-3 identical or different R atoms. 6 replace;

[0014] R 6 The following groups are represented by hydrogen atom, hydroxyl group, halogen, cyano group, mercapto group, carboxyl group, carbamoyl group, substituted or unsubstituted (C1-C6) alkyl group, substituted or unsubstituted (C1-C6) alkoxy group, substituted or unsubstituted (C1-C6) alkylthio group, substituted or unsubstituted (C2-C6) alkenyl group, substituted or unsubstituted (C1-C6) alkylsulfinyl group, substituted or unsubstituted (C1-C6) alkylsulfonyl group, substituted or unsubstituted (C1-C6) alkylformyl group, substituted or unsubstituted (C1-C6) alkoxyformyl group, -CO-NHR 7 -CO-NR 7 R 8 -NH-CO-R 7 -NHR 7 or -NR 7 R 8 Among them, R 7 R 8 The substituted or unsubstituted (C1-C6) alkyl groups are the same or different; the substituents of each of the above substitutions are selected from hydroxyl, amino or halogen; the substitution is monosubstituted or polysubstituted, and the substituents of polysubstituted groups are the same or different;

[0015] R 4 It can be hydrogen, halogen, (C1-C4)alkyl, or (C1-C4)alkoxy.

[0016] m is 0, 1, or 2;

[0017] n is 2, 3, or 4.

[0018] This invention preferably relates to 2-substituted aminobenzothiazole compounds of general formula I and their pharmaceutically acceptable salts.

[0019] in,

[0020] When Y represents N atom, X represents S atom; when Y represents S atom, X represents N atom.

[0021] R 1 and R 2 Together with the nitrogen atoms they are attached to, they form 4-10 membered heterocyclic groups, which, in addition to the nitrogen atom, optionally contain 0-3 heteroatoms selected from N, O, or S. The heterocyclic groups may also optionally be bonded by 1-3 identical or different R atoms. 5 Substitution: Heterocyclic groups contain 0-2 carbon-carbon double bonds;

[0022] R 5 The groups are hydrogen atom, =O, hydroxyl, amino, halogen, carboxyl, cyano, nitro, (C1-C4)alkyl, (C1-C4)alkoxy, (C3-C6)cycloalkyl, (C1-C4)alkylsulfinyl, (C1-C4)alkylsulfonyl, and (C1-C4)alkylformyl.

[0023] R 3 It is a phenyl, a 5-6-membered heteroaryl, or an adamantyl group, wherein the heteroaryl group contains 1-3 heteroatoms selected from N, O, or S, and the phenyl or heteroaryl group may optionally be surrounded by 1-3 identical or different R atoms. 6 replace;

[0024] R 6 The following groups are represented by hydrogen atom, hydroxyl group, halogen, cyano group, mercapto group, carboxyl group, carbamoyl group, substituted or unsubstituted (C1-C4) alkyl group, substituted or unsubstituted (C1-C4) alkoxy group, substituted or unsubstituted (C1-C4) alkylthio group, substituted or unsubstituted (C2-C4) alkenyl group, substituted or unsubstituted (C1-C4) alkylsulfinyl group, substituted or unsubstituted (C1-C4) alkylsulfonyl group, substituted or unsubstituted (C1-C4) alkylformyl group, substituted or unsubstituted (C1-C4) alkoxyformyl group, -CO-NHR 7 -CO-NR 7 R 8 -NH-CO-R 7 -NHR 7 or -NR 7 R 8 Among them, R 7 R 8 The substituted or unsubstituted (C1-C4) alkyl groups are the same or different; the substituents of each of the above substitutions are selected from hydroxyl, amino or halogen; the substitution is monosubstituted or polysubstituted, and the substituents of polysubstituted groups are the same or different;

[0025] R 4 It can be hydrogen, halogen, or methyl;

[0026] m is 0 or 1;

[0027] n is 2 or 3.

[0028] The present invention more preferably relates to 2-substituted aminobenzothiazole compounds of general formula I and pharmaceutically acceptable salts thereof, wherein,

[0029] When Y represents N atom, X represents S atom; when Y represents S atom, X represents N atom.

[0030] R 1 and R 2 Together with the nitrogen atoms they are attached to, they form

[0031] R 3 It is phenyl or adamantyl; the phenyl group is optionally surrounded by 1-3 identical or different R groups. 6 replace;

[0032] R 6 Halogen, unsubstituted or optionally substituted with 1-3 identical or different halogens (C1-C4) alkyl or (C1-C4) alkoxy;

[0033] R 4 It can be hydrogen, fluorine, or chlorine;

[0034] m is 0 or 1;

[0035] n is 2.

[0036] This invention particularly preferably relates to 2-substituted aminobenzothiazole compounds of general formula I and their pharmaceutically acceptable salts.

[0037] in,

[0038] When Y represents N atom, X represents S atom; when Y represents S atom, X represents N atom, and R 4 For H;

[0039] R 1 and R 2 Together with the nitrogen atoms they are attached to, they form

[0040] R 3 It is a phenyl group, and the para position of the phenyl group is R. 6 replace;

[0041] R 6 It is a halogen, trifluoromethyl or trifluoromethoxy;

[0042] R4 It can be hydrogen, fluorine, or chlorine;

[0043] m is 0 or 1;

[0044] n is 2.

[0045] This invention particularly preferably relates to 2-substituted aminobenzothiazole compounds of general formula II and pharmaceutically acceptable salts thereof, wherein,

[0046] R 1 and R 2 Together with the nitrogen atoms they are attached to, they form

[0047] R 3 It is a phenyl group, wherein the para position of the phenyl group is R 6 replace;

[0048] R 6 It is a halogen, trifluoromethyl or trifluoromethoxy;

[0049] R 4 It can be hydrogen, fluorine, or chlorine;

[0050] m is 0 or 1;

[0051] n is 2.

[0052] More preferably, the present invention provides the following 2-substituted aminobenzothiazole compounds and their pharmaceutically acceptable salts:

[0053] 1-{N-[2-(4-morpholinyl)ethyl]-2-aminobenzo[d]thiazolyl-5-yl}-3-(4-chlorophenyl)urea;

[0054] 1-{N-[2-(2-oxo-1-pyrrolidinyl)ethyl]-2-aminobenzo[d]thiazolyl-5-yl}-3-(4-chlorophenyl)urea;

[0055] 1-{N-[2-(4-morpholinyl)ethyl]-2-aminobenzo[d]thiazolyl-6-yl}-3-(4-chlorophenyl)urea;

[0056] 1-{N-[2-(dimethylamino)ethyl]-2-aminobenzo[d]thiazolyl-6-yl}-3-(4-chlorophenyl)urea;

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

[0058] 1-{N-[2-(1-piperidinyl)ethyl]-2-aminobenzo[d]thiazolyl-5-yl}-3-(4-chlorophenyl)urea;

[0059] 1-{N-[2-(4-morpholinyl)ethyl]-2-aminobenzo[d]thiazolyl-6-yl}-3-(4-trifluoromethoxyphenyl)urea;

[0060] 1-{N-[2-(2-oxo-1-pyrrolidinyl)ethyl]-2-aminobenzo[d]thiazolyl-5-yl}-3-(3-fluoro-4-chlorophenyl)urea;

[0061] 1-{N-[2-(4-morpholinyl)ethyl]-2-aminobenzo[d]thiazolyl-6-yl}-3-(1-adamantyl)urea;

[0062] 1-{N-[2-(2-oxo-1-pyrrolidinyl)ethyl]-2-aminobenzo[d]thiazolyl}-3-(4-chlorophenyl)urea;

[0063] 1-{N-[2-(3-oxo-4-morpholinyl)ethyl]-2-aminobenzo[d]thiazolyl-5-yl}-3-(3,4-dichlorophenyl)urea;

[0064] 1-{N-[2-(3-oxo-4-morpholinyl)ethyl]-2-amino-5-fluorobenzo[d]thiazolyl-6-yl}-3-(4-chlorophenyl)urea;

[0065] 1-{N-[2-(2-oxo-1-piperidinyl)ethyl]-2-amino-5-fluorobenzo[d]thiazolyl-6-yl}-3-(4-chlorophenyl)urea;

[0066] 1-{N-[2-(3-oxo-4-morpholinyl)ethyl]-2-aminobenzo[d]thiazo-6-yl}-3-(1-adamantyl)urea;

[0067] 1-{N-[2-(3-oxo-4-morpholinyl)ethyl]-2-amino-5-chlorobenzo[d]thiazolyl-6-yl}-3-(4-chlorophenyl)urea;

[0068] 1-{N-[2-(2-oxo-1-pyrrolidinyl)ethyl]-2-amino-5-fluorobenzo[d]thiazolyl-6-yl}-3-(4-chlorophenyl)urea.

[0069] Optionally, pharmaceutically acceptable salts of 2-substituted aminobenzothiazole compounds are salts formed by 2-substituted aminobenzothiazole compounds and acids 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.

[0070] This invention also includes prodrugs of the compounds of this invention. The prodrugs of the compounds of this invention are compounds of general formula I, which may have weak or no activity on their own, but are converted into the corresponding biologically active form under physiological conditions (e.g., through metabolism, solvation or other means) after administration.

[0071] Unless otherwise specified, the term "halogen" as used in this invention refers to fluorine, chlorine, or bromine. "Hydroxy" refers to -OH. "Amino" refers to -NH₂. "Carboxyl" refers to -COOH. "Cyano" refers to -CN. "Nitro" refers to -NO₂. "Carbamoyl" refers to -CO-NH₂.

[0072] "C1-C4" refers to the number of carbon atoms in the defined group (such as alkyl, alkoxy, cycloalkyl, etc.) being 1, 2, 3, or 4. The meanings of other terms described in a similar manner, such as "C1-C6" and "C3-C6", can be inferred from this.

[0073] "Alkyl" refers to a straight-chain or branched alkyl group. For example, "C1-C6 alkyl" refers to a straight-chain or branched alkyl group having 1 to 6 carbon atoms, such as methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, pentyl, and hexyl. "Alkenyl" refers to a straight-chain or branched alkenyl group.

[0074] "Cycloalkyl" refers to substituted or unsubstituted cycloalkyl groups. For example, "C3-C6 cycloalkyl" refers to cycloalkyl groups having 3 to 6 carbon atoms, such as cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

[0075] "Alkoxy" is an -OR group, "alkylthio" is an -SR group, "alkylsulfinyl" is an -SO-R group, "alkylsulfonyl" is an -SO2-R group, "alkylformyl" is an -CO-R group, and "alkoxyformyl" is an -CO-OR group, where R is an alkyl group as defined above.

[0076] "Heteroaryl" refers to a cyclic system containing one or more monocyclic or polycyclic heteroatoms selected from N, O, and S. The cyclic system is aromatic, such as imidazolyl, pyridinyl, pyrazolyl, furanyl, thiophene, pyrrololyl, thiazolyl, benzothiazolyl, oxazolyl, isoxazolyl, quinolinyl, isoquinolinyl, benzimidazolyl, benzoxazolyl, etc. "Heterocyclic" refers to a cyclic system containing one or more monocyclic or polycyclic heteroatoms selected from N, O, and S, such as pyrroloalkyl, morpholinyl, piperazinyl, piperidinyl, pyrazolalkyl, imidazoalkyl, and thiazolyl, etc.

[0077] The present invention also provides a pharmaceutical composition comprising one or more of the above-described 2-substituted aminobenzothiazole compounds and their pharmaceutically acceptable salts, as well as a pharmaceutically acceptable carrier. The compounds of the present invention can be used in combination with other active ingredients, provided they do not produce other adverse effects, such as allergic reactions.

[0078] The carriers used in the pharmaceutical compositions of this invention are common types available in the pharmaceutical field, including: binders, lubricants, disintegrants, solubilizers, diluents, stabilizers, suspending agents, pigments, flavoring agents, etc., for oral formulations; pH adjusters, osmotic pressure adjusters, solubilizers, stabilizers, etc., for injectable formulations; and matrices, diluents, lubricants, preservatives, etc., for topical formulations. The pharmaceutical formulations can be administered orally, parenterally (e.g., intravenously, subcutaneously, intraperitoneally, etc.), or topically (e.g., through the eyes, nose, sublingually, skin, etc.). If certain drugs are unstable under gastric conditions, they can be formulated into enteric-coated tablets.

[0079] In vitro and in vivo inhibitory activity tests revealed that the compound of general formula I and its pharmaceutically acceptable salt exhibit significant inhibitory activity against sEH, stabilizing the level of EETs in vivo. Pharmacodynamic studies showed that the compounds of this invention can inhibit carrageenan-induced foot edema, protect against ischemic brain injury, maintain glycemic homeostasis, relieve neuropathic pain, resist tissue fibrosis, alleviate depression, and improve dry eye symptoms. They can be used to treat sEH-mediated inflammatory diseases, cardiovascular and cerebrovascular diseases, diabetes and its complications, fibrotic diseases, neurological and psychiatric disorders, pain, ulcerative diseases, or dry eye syndrome.

[0080] The precise amount of the compounds of this invention required to treat sEH-mediated conditions will vary from person to person, depending on the type of patient, age and general condition, severity of the disease being treated, the specific compound used, and the route of administration, such as the route and frequency of administration. Those skilled in the art can determine the appropriate effective amount using only conventional experimental methods.

[0081] The dosage of the compound can range from about 0.1 to 100 mg / kg body weight / day, preferably 1 to 50 mg / kg body weight / day. It is understood that the dosage may vary depending on the patient's needs, the severity of the inflammatory disease, cardiovascular disease, diabetes, diabetic complications, diabetes-related diseases, fibrotic diseases, neurological and psychiatric disorders, pain, ulcerative diseases, or dry eye syndrome being treated, and the specific compound used. Furthermore, it is understood that the initial dose may be increased beyond the upper limit to rapidly achieve the desired blood level, or the initial dose may be lower than the optimal value, and the daily dose may be gradually increased during treatment, depending on the specific circumstances. If necessary, the daily dose may also be divided into multiple administrations, such as 2-4 times daily.

[0082] Mammals refer to humans or animals.

[0083] The amount of the active ingredient, i.e., the compound according to the invention, in the pharmaceutical composition and its unit dosage form can vary, depending on the specific application, the potency of the specific compound, and the required concentration. Generally, the content of the active ingredient will be between 0.5% and 90% by total weight of the composition.

[0084] In combination therapy, the compounds of the present invention and other compounds may be administered simultaneously or at intervals. When administered simultaneously, the compounds of the present invention and other compounds may be combined in a single pharmaceutical composition or in separate compositions.

[0085] This invention also proposes the use of compounds of general formula I and their pharmaceutically acceptable salts or pharmaceutical compositions in the preparation of medicaments for the treatment and / or prevention of sEH-mediated diseases. These diseases include inflammatory diseases, cardiovascular and cerebrovascular diseases, diabetes, diabetic complications, diabetes-related diseases, fibrotic diseases, neurological and psychiatric diseases, pain, ulcerative diseases, and dry eye syndrome.

[0086] Specifically, inflammatory diseases include inflammatory liver disease, inflammatory kidney disease, inflammatory lung disease, inflammatory brain disease, myocarditis, pancreatitis, arthritis, soft tissue inflammation, osteomyelitis, and vascular inflammation; cardiovascular and cerebrovascular diseases include hypertension, myocardial infarction, heart failure, coronary heart disease, cardiovascular arteriosclerosis, ischemic stroke, and hemorrhagic stroke; diabetes includes type 1 or type 2 diabetes; diabetic complications include diabetic retinopathy, diabetic uveitis, diabetic cataracts, diabetic nephropathy, diabetic skin diseases, and diabetic peripheral neuropathy; and diabetes-related diseases include hyperlipidemia, high cholesterol, and high blood sugar. Hyperuricemia and gout, obesity and metabolic syndrome; fibrotic diseases including pulmonary fibrosis, liver fibrosis, myocardial fibrosis and renal fibrosis; neurological and psychiatric diseases including Alzheimer's disease, epilepsy, Parkinson's disease, amyotrophic lateral sclerosis, schizophrenia, mental disorders, depression, neurasthenia; pain diseases including neuropathic pain, inflammatory pain, tumor pain and mixed pain; ulcerative diseases including gastric ulcer, duodenal ulcer, ulcerative colitis, corneal ulcer and oral ulcer; dry eye syndrome including aqueous hypoplasia dry eye, lipid abnormality dry eye, mucin abnormality dry eye, tear dynamics abnormality dry eye and mixed dry eye.

[0087] This invention relates to a method of treating and / or preventing sEH-mediated diseases by administering a compound of general formula I and a pharmaceutically acceptable salt thereof to a subject in need. The compounds of general formula I and pharmaceutically acceptable salts thereof of this invention exhibit sEH inhibitory activity and are therefore effective for the prevention or treatment of sEH-mediated related diseases.

[0088] In this invention, the term "prevention" refers to any effect of inhibiting or delaying the occurrence, spread, and recurrence of sEH-mediated related diseases due to the application of the compounds or compositions of this invention, and the term "treatment" refers to all effects of alleviating or beneficially altering the symptoms of the aforementioned diseases by applying the compounds or compositions of this invention.

[0089] The examples and preparation methods provided below further illustrate and demonstrate the compounds of the present invention and their preparation methods. It should be understood that the scope of the following examples and preparation methods does not limit the scope of the present invention in any way.

[0090] The following synthetic routes describe the preparation of compounds of general formula I of the present invention. All starting materials are prepared by the methods described in these synthetic routes, by methods well known to those skilled in the art of organic chemistry, or are commercially available. All final compounds of the present invention are prepared by the methods described in these synthetic routes or by similar methods well known to those skilled in the art of organic chemistry. All variables used in these synthetic routes are defined below or as defined in the claims.

[0091] The synthetic route for compounds of general formula I is as follows: starting material A reacts with Boc2(O) to obtain intermediate B, and intermediate B reacts with R... 1 R 2 N(CH2) n Cl undergoes a nucleophilic substitution reaction to give intermediate C, which is then reduced by ferric acid or by H2 / Pd-C to give intermediate D; R 3 (CH2) m NH2 reacts with phenyl chloroformate to prepare intermediate E; intermediate D and intermediate E undergo a condensation reaction to prepare intermediate F; intermediate F is deprotected by the Boc protecting group under the action of hydrogen chloride to obtain the hydrochloride salt of compound of general formula I; the latter is then reacted with sodium hydroxide solution to obtain the free compound of general formula I. Attached Figure Description

[0092] Figure 1 shows the anti-inflammatory effect of the compounds in the embodiments of the present invention. # Compared with the solvent group, p<0.05; ## compared with the solvent group, p<0.01; ** compared with the celecoxib group, p<0.01.

[0093] Figure 2 shows the effect of the compound of the present invention in reducing the infarct volume in rats with cerebral ischemia. * p < 0.05 compared with the solvent group, # p < 0.05 compared with the edaravone group, $ p < 0.05 compared with the t-AUCB group.

[0094] Figure 3 shows the effect of the compound of the present invention in reducing the level of inflammatory factors in the brain tissue of rats with cerebral ischemia. * p < 0.05 compared with the solvent group, # p < 0.05 compared with the sham-operated group, $ p < 0.05 compared with the t-AUCB group.

[0095] Figure 4 shows the effect of the compound in the embodiment of the present invention on improving the survival rate of rats with cerebral ischemia.

[0096] Figure 5 shows the effect of the compound of the present invention on improving the learning and memory function of rats with cerebral ischemia. * Compared with the solvent group, p<0.05; # Compared with the sham-operated group, p<0.05; $ Compared with the t-AUCB group, p<0.05; & Compared with the edaravone group, p<0.05.

[0097] Figure 6 shows the hypoglycemic effect of the compounds in the embodiments of the present invention. * 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;

[0098] Figure 7 shows the anti-fibrotic effect of the compounds in the embodiments of the present invention. * p < 0.05 compared with the solvent group, # p < 0.05 compared with the sham surgery group, $ p < 0.05 compared with the t-AUCB group;

[0099] Figure 8 shows the analgesic effect of the compound of the present invention on selective injury of sciatic nerve branches. * p < 0.05 compared with the solvent group, # p < 0.05 compared with the EC5026 group;

[0100] Figure 9 shows the analgesic effect of the compounds in the embodiments of the present invention on diabetic pathological neuropathic pain. * p < 0.05 compared with the solvent group, # p < 0.05 compared with the pregabalin group;

[0101] Figure 10 shows the analgesic effect of the compounds in the embodiments of the present invention on paclitaxel-induced neuropathic pain. * p < 0.05 compared with the solvent group, # p < 0.05 compared with the pregabalin group;

[0102] Figure 11 shows the effect of the compound of the present invention in reducing the level of inflammatory factors in the spinal cord tissue of animals with neuropathic pain. * p < 0.05 compared with the solvent group, ## p < 0.01 compared with the sham surgery group, $ p < 0.05 compared with the EC5026 group.

[0103] Figure 12 shows the effect of the compound of the present invention on the motor coordination function of rats, **p<0.01 compared with before drug administration;

[0104] Figure 13 shows the antidepressant effect of the compounds in the embodiments of the present invention. * p < 0.05 compared with the solvent group, # p < 0.05 compared with the normal group;

[0105] Figure 14 shows the effect of the compound in the embodiment of the present invention on treating dry eye syndrome. ** p < 0.01 compared with the solvent group, ## p < 0.01 compared with the normal group, $ p < 0.05 compared with the cyclosporine eye drops group;

[0106] Figure 15 shows the effect of the compounds of the present invention on improving the levels of 14,15-EET and 14,15-DHET in the corneal tissue of mice with dry eye syndrome. * p < 0.05 compared with the solvent group, # p < 0.05 compared with the normal group, $ p < 0.05 compared with the cyclosporine eye drops group.

[0107] Figure 16 shows the effect of the compound of the present invention in reducing the level of inflammatory factors in the corneal tissue of mice with dry eye syndrome. * p < 0.05 compared with the solvent group, # p < 0.05 compared with the normal group, $ p < 0.05 compared with the cyclosporine eye drops group.

[0108] Figure 17 shows the effect of the compound of the present invention in reducing retinal vascular permeability in 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.

[0109] Figure 18 shows the effect of the compound of the present invention on protecting retinal vascular homeostasis in 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 pericytes, ▲ represents cellless capillaries;

[0110] Figure 19 shows the distribution results of the compounds of the present invention in Dutch rabbit eye tissue;

[0111] Figure 20 shows the effect of the compounds of the present invention on the levels of 14,15-EET and 14,15-DHET in rats. *Compared with the EC5026 group, p<0.05;

[0112] Figure 21 shows the molecular docking results of EC5026 and sEH;

[0113] Figure 22 shows the molecular docking results of the compound in Example 1 of this invention with sEH;

[0114] Figure 23 shows the molecular docking results of the compound in Example 3 of this invention with sEH;

[0115] Figure 24 shows the molecular docking results of compound 16 of this invention with sEH. Detailed implementation method:

[0116] The following examples depict methods for preparing some of the compounds described herein. It should be understood that the following methods, as well as other methods known to those skilled in the art, are applicable to the preparation of all the compounds described herein. The examples are intended to illustrate, but not limit, the scope of the invention.

[0117] This invention relates to a series of 2-substituted aminobenzothiazole compounds, all of which exhibit significant inhibitory activity in vitro according to sEH inhibition tests. In vivo pharmacodynamic studies have shown that they have good therapeutic effects on inflammation, stroke, diabetes, fibrosis, pain, depression, and dry eye syndrome.

[0118] 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 1100 LC / MSD; all reagents used were of analytical or chemical purity.

[0119] Example 1 Preparation of 1-{N-[2-(4-morpholinyl)ethyl]-2-aminobenzo[d]thiazolyl-5-yl}-3-(4-chlorophenyl)urea

[0120] 1.1 Preparation of N-tert-butoxycarbonyl-2-amino-5-nitrobenzo[d]thiazole (B)

[0121] 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%.

[0122] ESI-MS m / z: 296.1 [M+H] + .

[0123] 1.2 Preparation of N-[2-(4-morpholinyl)ethyl]-N-tert-butoxycarbonyl-2-amino-5-nitrobenzo[d]thiazole (C)

[0124] 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%.

[0125] ESI-MS m / z: 443.1 [M+Na] + .

[0126] 1.3 N 2 -[2-(4-morpholinyl)ethyl]-N 2 Preparation of -tert-butoxycarbonyl-2,5-diaminobenzo[d]thiazole (D)

[0127] At room temperature, 2.3 g (41.1 mmol) of reduced iron powder and 4.4 g (82.4 mmol) of ammonium chloride were added to 45 mL of 95% ethanol and refluxed for 10 min. 4.2 g (10.3 mmol) of intermediate C was added in portions to the ethanol solution, and the mixture was refluxed for 2 h after the addition was complete. After the reaction was complete, the mixture was filtered while hot, and the filtrate was evaporated to dryness under reduced pressure to give 2.5 g of a brown solid, with a yield of 65.2%.

[0128] ESI-MS m / z: 379.1 [M+H] + .

[0129] 1.4 Synthesis of N-(4-chlorophenyl)carbamate (E)

[0130] Under ice-water bath conditions, 1.4 g (11.2 mmol) of 4-chloroaniline and 0.7 g (6.7 mmol) of sodium carbonate were added to a tetrahydrofuran solution, and the mixture was stirred. Then, 1.9 g (11.9 mmol) of a tetrahydrofuran solution of phenyl chloroformate (3 mL) was added dropwise to the reaction mixture. After the addition was complete, the reaction mixture was allowed to react at room temperature for 2 h. After the reaction was complete, the reaction mixture was evaporated to dryness under reduced pressure. 10 mL of water was added to the residue, and the mixture was stirred. A solid precipitated, which was then filtered. The filter cake was washed with water and dried to obtain 2.5 g of solid, with a yield of 91%.

[0131] 1.5 Preparation of 1-{N-[2-(4-morpholinyl)ethyl]-N-tert-butoxycarbonyl-2-aminobenzo[d]thiazolyl-5-yl}-3-(4-chlorophenyl)urea (F)

[0132] At room temperature, 2.0 g (5.3 mmol) of intermediate D and 0.8 mL (5.8 mmol) of triethylamine were added to 20 mL of dry dioxane and stirred until dissolved. 1.6 g (6.3 mmol) of intermediate E was added in portions to the reaction mixture. After the addition was complete, the reaction mixture was heated to 55 °C and reacted for 5 h. After the reaction was complete, the reaction mixture was cooled to room temperature, 20 mL of water was added, the mixture was stirred, filtered, and dried to give 2.2 g of a white solid, with a yield of 78.6%.

[0133] 1.6 Preparation of 1-{N-[2-(4-morpholinyl)ethyl]-2-aminobenzo[d]thiazolyl-5-yl}-3-(4-chlorophenyl)urea (Example 1)

[0134] At room temperature, 0.19 g (0.36 mmol) of intermediate F was added to 2 mL of a saturated dioxane solution containing hydrogen chloride, and the reaction solution was heated to 60 °C and reacted for 3 h. After the reaction was complete, the reaction solution was cooled to room temperature and filtered to obtain 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 a 20% sodium hydroxide aqueous solution. A solid precipitated out, which was filtered and dried to obtain 0.12 g of a white solid, which was the compound of Example 1, with a yield of 79.0%.

[0135] 1 H NMR(600MHz,CD3OD)δ7.76(s,1H),7.50(d,J=8.5Hz,1H),7.44(d,J=8.8Hz,2H),7.27(d, J=8.8Hz,2H),7.09(dd,J=8.5,1.9Hz,1H),3.87(m,4H),3.72(m,2H),3.07(m,6H).ESI-MS m / z:431.9[M+H] + .

[0136] With suitable R 4 Using substituted 2-amino-5-(6-)nitrobenzo[d]thiazole (A) as a starting material, the key intermediate D was synthesized according to the synthesis methods in Examples 1.1 to 1.3; with R 3 Using substituted aniline as a starting material, intermediate E was synthesized according to the synthesis method in 1.4 of Example 1; intermediates D and E were synthesized according to the synthesis method in 1.5 of Example 1 to obtain intermediate F, and then the compounds of Examples 2 to 16 were synthesized according to the synthesis method in 1.6. Their structural formulas are shown in Table 1.

[0137] Example 2 Preparation of 1-{N-[2-(2-oxo-1-pyrrolidinyl)ethyl]-2-aminobenzo[d]thiazolyl}-3-(4-chlorophenyl)urea

[0138] Using 2-amino-5-nitrobenzo[d]thiazole as a starting material, key intermediate B was synthesized according to the synthesis method in 1.1 of Example 1; intermediate B and N-(2-chloroethyl)pyrrolidine-2-one were synthesized according to the synthesis methods in 1.2 to 1.3 of Example 1 to obtain intermediate D; using 4-chloroaniline as a starting material, intermediate E was synthesized according to the synthesis method in 1.4 of Example 1 to obtain intermediate E; intermediate D and intermediate E were synthesized according to the synthesis methods in 1.5 to 1.6 of Example 1 to obtain the compound of Example 2.

[0139] 1 H NMR (400MHz, CD3OD) δ7.62(d,J=2.0Hz,1H),7.47(d,J=8.5Hz,1H),7.43(d,J=8.9Hz,2H),7.27(d,J=8.9Hz,2H),7.1 4(dd,J=8.5,2.1Hz,1H),3.64(m,2H),3.57-3.53(m,4H),2.32(t,J=8.1Hz,2H),2.0(m,2H).MS(ESI)m / z:428.0[MH] - .

[0140] Example 3 Preparation of 1-{N-[2-(4-morpholinyl)ethyl]-2-aminobenzo[d]thiazolyl-6-yl}-3-(4-chlorophenyl)urea

[0141] Using 2-amino-6-nitrobenzo[d]thiazole as a starting material, key intermediate D was synthesized according to the synthesis methods 1.1 to 1.3 in Example 1; using 4-chloroaniline as a starting material, intermediate E was synthesized according to the synthesis method 1.4 in Example 1; intermediates D and E were synthesized according to the synthesis methods 1.5 to 1.6 in Example 1 to obtain the compound of Example 3.

[0142] 1 H NMR (400MHz, CD3OD) δ7.84(d,J=2.2Hz,1H),7.42(d,J=8.9Hz,2H),7.35(d,J=8.6Hz,1H),7.26(d,J=8.9Hz,2H),7.15(dd,J =8.6,2.2Hz,1H),3.73(t,J=4.7Hz,4H),3.59(t,J=6.4Hz,2H),2.71(t,J=5.8Hz,2H),2.60(m,4H).MS(ESI)m / z:430.0[MH] - .

[0143] Example 4 Preparation of 1-{N-[2-(dimethylamino)ethyl]-2-aminobenzo[d]thiazolyl}-3-(4-chlorophenyl)urea

[0144] Using 2-amino-6-nitrobenzo[d]thiazole as a starting material, key intermediate B was synthesized according to the synthesis method in 1.1 of Example 1; intermediate B and N,N-dimethyl-2-chloroethylamine were used to synthesize intermediate D according to the synthesis methods in 1.2 to 1.3 of Example 1; using 4-chloroaniline as a starting material, intermediate E was synthesized according to the synthesis method in 1.4 of Example 1; intermediate D and intermediate E were used to prepare the compound of Example 4 according to the synthesis methods in 1.5 to 1.6 of Example 1.

[0145] MS(ESI) m / z: 390.0 [M+H] + .

[0146] Example 5 Preparation of 1-{N-[2-(4-morpholinyl)ethyl]-2-aminobenzo[d]thiazolyl-5-yl}-3-(3-fluoro-4-chlorophenyl)urea

[0147] Using 2-amino-5-nitrobenzo[d]thiazole as a starting material, key intermediate B was synthesized according to the synthesis method in 1.1 of Example 1; intermediate B and N-(2-chloroethyl)piperidine were synthesized into intermediate D according to the synthesis methods in 1.2 to 1.3 of Example 1; using 3-fluoro-4-chloroaniline as a starting material, intermediate E was synthesized according to the synthesis method in 1.4 of Example 1; intermediate D and intermediate E were synthesized according to the synthesis methods in 1.5 to 1.6 of Example 1 to obtain the compound of Example 5.

[0148] 1 H NMR (400MHz, CD3OD) δ7.61-7.58(m,2H),7.48(d,J=8.5Hz,1H),7.33(t,J=8.5Hz,1H),7.14(dd,J=8.5,2.1Hz,1H ),7.09(m,1H),3.71(t,J=4.6Hz,4H),3.58(t,J=6.5Hz,2H),2.66(t,J=6.5Hz,2H),2.54(t,J=4.5Hz,4H).ESI-MS m / z:450.5[M+H] + .

[0149] Example 6 Preparation of 1-{N-[2-(1-piperidinyl)ethyl]-2-aminobenzo[d]thiazolyl}-3-(4-chlorophenyl)urea

[0150] Using 2-amino-5-nitrobenzo[d]thiazole as a starting material, key intermediate D was synthesized according to the synthesis methods 1.1 to 1.3 in Example 1; using 3-fluoro-4-chloroaniline as a starting material, intermediate E was synthesized according to the synthesis method 1.4 in Example 1; intermediates D and E were synthesized according to the synthesis methods 1.5 to 1.6 in Example 1 to obtain the compound of Example 6.

[0151] 1 H NMR (400MHz, CD3OD) δ7.65(s,1H),7.49(d,J=8.5Hz,1H),7.43(d,J=8.8Hz,2H),7.27(d,J=8.8Hz,2H),7.1 2(dd,J=8.5,2.0Hz,1H),3.63(t,J=6.4Hz,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] + .

[0152] Example 7 Preparation of 1-{N-[2-(4-morpholinyl)ethyl]-2-aminobenzo[d]thiazolyl-6-yl}-3-(4-trifluoromethoxyphenyl)urea

[0153] Using 2-amino-6-nitrobenzo[d]thiazole as a starting material, key intermediate D was synthesized according to the synthesis methods in 1.1 to 1.3 of Example 1; using 4-(trifluoromethoxy)aniline as a starting material, intermediate E was synthesized according to the synthesis method in 1.4 of Example 1; intermediates D and E were synthesized according to the synthesis methods in 1.5 to 1.6 of Example 1 to obtain the compound of Example 7.

[0154] ESI-MS m / z: 481.9 [M+H] + .

[0155] Example 8 Preparation of 1-{N-[2-(2-oxo-1-pyrrolidinyl)ethyl]-2-aminobenzo[d]thiazolyl}-3-(3-fluoro-4-chlorophenyl)urea

[0156] Using 2-amino-5-nitrobenzo[d]thiazole as a starting material, key intermediate B was synthesized according to the synthesis method in 1.1 of Example 1; intermediate B and N-(2-chloroethyl)pyrrolidine-2-one were synthesized according to the synthesis methods in 1.2 to 1.3 of Example 1 to obtain intermediate D; using 3-fluoro-4-chloroaniline as a starting material, intermediate E was synthesized according to the synthesis method in 1.4 of Example 1 to obtain intermediate E; intermediate D and intermediate E were synthesized according to the synthesis methods in 1.5 to 1.6 of Example 1 to obtain the compound of Example 8.

[0157] 1H NMR (600MHz, DMSO-d6) δ9.11(s,1H),8.86(s,1H),8.15(s,1H),7.69(d,J=11.7Hz,1H),7.62(s,1H),7.55(d,J=8.0Hz,1H),7.47(t,J=8 .4Hz,1H),7.19(d,J=8.4Hz,1H),7.09(d,J=8.1Hz,1H),3.51(s,4H),3.43(m,4H),2.18(t,J=7.3Hz,2H),1.90(t,J=7.0Hz,2H).ESI-MS m / z:448.1[M+H] + .

[0158] Example 9 Preparation of 1-{N-[2-(4-morpholinyl)ethyl]-2-aminobenzo[d]thiazolyl-6-yl}-3-(1-adamantyl)urea

[0159] Using 2-amino-6-nitrobenzo[d]thiazole as a starting material, key intermediate D was synthesized according to the synthesis methods 1.1 to 1.3 in Example 1; using adamantane as a starting material, intermediate E was synthesized according to the synthesis method 1.4 in Example 1; intermediates D and E were synthesized according to the synthesis methods 1.5 to 1.6 in Example 1 to obtain the compound of Example 9.

[0160] ESI-MS m / z: 456.0 [M+H] + 477.9 [M+Na] + .

[0161] Example 10 Preparation of 1-{N-[2-(2-oxo-1-pyrrolidinyl)ethyl]-2-aminobenzo[d]thiazo-6-yl}-3-(4-chlorophenyl)urea

[0162] Using 2-amino-6-nitrobenzo[d]thiazole as a starting material, key intermediate B was synthesized according to the synthesis method in 1.1 of Example 1; intermediate B and N-(2-chloroethyl)pyrrolidine-2-one were synthesized according to the synthesis methods in 1.2 to 1.3 of Example 1 to obtain intermediate D; using 4-chloroaniline as a starting material, intermediate E was synthesized according to the synthesis method in 1.4 of Example 1; intermediate D and intermediate E were synthesized according to the synthesis methods in 1.5 to 1.6 of Example 1 to obtain the compound of Example 10.

[0163] 1H NMR(600MHz,DMSO-d6)δ10.33(br s,1H),9.54(s,1H),9.51(s,1H),8.10(d,J=1.7Hz,1H),7.52(d,J=8.7Hz,1H),7.49(d,J=8.8Hz,2H),7.43(dd,J=8.7,1.9Hz,1 H),7.33(d,J=8.8Hz,2H),3.71(m,2H),3.48(t,J=5.9Hz,2H),3.44(t,J=7.0Hz,2H),2.19(t,J=8.0Hz,2H),1.92(m,2H).ESI-MS m / z:429.9[M+H] + .

[0164] Example 11 Preparation of 1-{N-[2-(3-oxo-4-morpholinyl)ethyl]-2-aminobenzo[d]thiazolyl-5-yl}-3-(3,4-dichlorophenyl)urea

[0165] Using 2-amino-5-nitrobenzo[d]thiazole as a starting material, key intermediate B was synthesized according to the synthesis method in 1.1 of Example 1; intermediate B and N-(2-chloroethyl)morpholin-3-one were synthesized according to the synthesis methods in 1.2 to 1.3 of Example 1 to obtain intermediate D; using 3,4-dichloroaniline as a starting material, intermediate E was synthesized according to the synthesis method in 1.4 of Example 1 to obtain intermediate E; intermediate D and intermediate E were synthesized according to the synthesis methods in 1.5 to 1.6 of Example 1 to obtain the compound of Example 11.

[0166] 1 HNMR (600MHz, DMSO-d6) δ9.11(s,1H),8.92(s,1H),8.31(brs,1H),7.89(d,J=2.0Hz,1H),7.63(s,1H),7.56(d,J=8.4Hz,1H),7.52(d,J=8. 8Hz,1H),7.34(dd,J=8.8,2.0Hz,1H),7.10(d,J=8.4Hz,1H),4.00(s,2H),3.79(t,J=4.9Hz,2H),3.56(m,4H),3.42(t,J=4.8Hz,2H).ESI-MS m / z:502.4[M+Na] + .

[0167] Example 12 Preparation of 1-{N-[2-(3-oxo-4-morpholinyl)ethyl]-2-amino-5-fluorobenzo[d]thiazolyl-6-yl}-3-(4-chlorophenyl)urea

[0168] Using 2-amino-5-fluoro-6-nitrobenzo[d]thiazole as a starting material, key intermediate B was synthesized according to the synthesis method in 1.1 of Example 1; intermediate B and N-(2-chloroethyl)morpholin-3-one were synthesized according to the synthesis methods in 1.2 to 1.3 of Example 1 to obtain intermediate D; using 4-chloroaniline as a starting material, intermediate E was synthesized according to the synthesis method in 1.4 of Example 1; intermediate D and intermediate E were synthesized according to the synthesis methods in 1.5 to 1.6 of Example 1 to obtain the compound of Example 12.

[0169] 1 H NMR (400MHz, DMSO-d6) δ9.31(s,1H),8.53(s,1H),8.30(d,J=7.9Hz,1H),8.21(s,1H),7.53(d,J=8.9Hz,2H),7.38(d, J=8.8Hz,2H),7.34(d,J=12.1Hz,1H),4.05(s,2H),3.84(t,J=4.8Hz,2H),3.59(m,4H),3.45(t,J=4.8Hz,2H).ESI-MS m / z:464.1[M+H] + .

[0170] Example 13 Preparation of 1-{N-[2-(2-oxo-1-piperidinyl)ethyl]-2-amino-5-fluorobenzo[d]thiazolyl-6-yl}-3-(4-chlorophenyl)urea

[0171] Using 2-amino-5-fluoro-6-nitrobenzo[d]thiazole as a starting material, key intermediate B was synthesized according to the synthesis method in 1.1 of Example 1; intermediate B and N-(2-chloroethyl)piperidin-2-one were synthesized according to the synthesis methods in 1.2 to 1.3 of Example 1 to obtain intermediate D; using 4-chloroaniline as a starting material, intermediate E was synthesized according to the synthesis method in 1.4 of Example 1; intermediate D and intermediate E were synthesized according to the synthesis methods in 1.5 to 1.6 of Example 1 to obtain the compound of Example 13.

[0172] 1 H NMR (400MHz, CD3OD) δ8.19(d,J=7.7Hz,1H),7.73-7.69(m,1H),7.63–7.60(m,1H),7.44(d,J=8.9Hz,2H),7.27(d,J=8.9H z,2H),7.19(d,J=11.9Hz,1H),3.65-3.60(m,4H),3.43(t,J=5.7Hz,2H),2.30(t,J=6.3Hz,2H),1.77-1.70(m,4H).ESI-MS m / z:462.1[M+H] + .

[0173] Example 14 Preparation of 1-{N-[2-(3-oxo-4-morpholinyl)ethyl]-2-aminobenzo[d]thiazo-6-yl}-3-(1-adamantyl)urea

[0174] Using 2-amino-6-nitrobenzo[d]thiazole as a starting material, key intermediate B was synthesized according to the synthesis method in 1.1 of Example 1; intermediate B and N-(2-chloroethyl)morpholin-3-one were synthesized according to the synthesis methods in 1.2 to 1.3 of Example 1 to obtain intermediate D; using adamantaneamine as a starting material, intermediate E was synthesized according to the synthesis method in 1.4 of Example 1; intermediate D and intermediate E were synthesized according to the synthesis methods in 1.5 to 1.6 of Example 1 to obtain the compound of Example 14.

[0175] ESI-MS m / z: 469.9 [M+H] + 491.9 [M+Na] + .

[0176] Example 15 Preparation of 1-{N-[2-(3-oxo-4-morpholinyl)ethyl]-2-amino-5-chlorobenzo[d]thiazo-6-yl}-3-(4-chlorophenyl)urea

[0177] Using 2-amino-5-chloro-6-nitrobenzo[d]thiazole as a starting material, key intermediate B was synthesized according to the synthesis method in 1.1 of Example 1; intermediate B and N-(2-chloroethyl)morpholin-3-one were synthesized according to the synthesis methods in 1.2 to 1.3 of Example 1 to obtain intermediate D; using 4-chloroaniline as a starting material, intermediate E was synthesized according to the synthesis method in 1.4 of Example 1; intermediate D and intermediate E were synthesized according to the synthesis methods in 1.5 to 1.6 of Example 1 to obtain the compound of Example 15.

[0178] 1 H NMR(400MHz,CD3OD)δ8.23(s,1H),7.46-7.44(m,3H),7.27(d,J=8.9Hz,2H),4 .07(s,2H),3.83(t,J=4.9Hz,2H),3.68(m,4H),3.50(t,J=5.2Hz,2H).ESI-MS m / z:501.8[M+Na] + .

[0179] Example 16 Preparation of 1-{N-[2-(2-oxo-1-pyrrolidinyl)ethyl]-2-amino-5-fluorobenzo[d]thiazolyl}-3-(4-chlorophenyl)urea

[0180] Using 2-amino-5-fluoro-6-nitrobenzo[d]thiazole as a starting material, key intermediate B was synthesized according to the synthesis method in 1.1 of Example 1; intermediate B and N-(2-chloroethyl)pyrrolidine-2-one were synthesized according to the synthesis methods in 1.2 to 1.3 of Example 1 to obtain intermediate D; using 4-chloroaniline as a starting material, intermediate E was synthesized according to the synthesis method in 1.4 of Example 1 to obtain intermediate E; intermediate D and intermediate E were synthesized according to the synthesis methods in 1.5 to 1.6 of Example 1 to obtain compound 16.

[0181] 1 H NMR(400MHz,DMF-d7)δ9.46(s,1H),8.68(d,J=2.1Hz,1H),8.60(d,J=7.9Hz,1H),8.26(t,J=5.7Hz,1H),7.80(d,J=8.9Hz,2H ),7.55(d,J=8.9Hz,2H),7.47(d,J=12.2Hz,1H),3.84(q,J=5.9Hz,2H),3.72–3.69(m,4H),3.10(m,2H),2.93(m,2H).ESI-MS m / z:447.9[M+H] + .

[0182] Table 1. Structural formulas of compounds from Examples 1-16

[0183] Bioactivity testing:

[0184] 1. Inhibitory activity tests of compounds in some examples against sEH

[0185] The in vitro inhibitory activity of 2-substituted aminobenzothiazole compounds of general formula I according to the present invention against sEH was tested using fluorescence analysis. t-AUCB was used as the reference standard.

[0186] Both the sEH solution and the fluorescent substrate PHOME (2-(3-phenylethyleneoxy-2-yl)acetic acid 1-cyano-1-(6-methoxy-2-naphthyl)methyl ester) solution were prepared with 25 mM Bis-Tris (pH 7.0) buffer. The compound and t-AUCB were dissolved and diluted with DMSO to prepare a series of different concentrations. 100 μL of enzyme solution and 5 μL of test compound solution were added to each well of a 96-well plate, and incubated at 37°C for 5 min. Then, 95 μL of substrate solution was added, and incubation continued at 37°C for 15 min. Blank wells contained only the corresponding volume of Bis-Tris (pH 7.0) buffer and DMSO, while full-well wells contained only sEH, substrate, and DMSO. After incubation, fluorescence intensity was measured at an excitation wavelength of 330 nm and an emission wavelength of 465 nm. The inhibition rate of different concentrations of the compound was calculated based on the full-well values, and the IC50 was calculated using SPSS software. 50 value.

[0187] IC50 of some of the compound examples and t-AUCB against sEH 50 The data is shown in Table 2.

[0188] Table 2 shows the inhibitory activity of some of the compounds in the examples against sEH.

[0189] Preliminary in vitro sEH inhibitory activity test results show that the compound of general formula I to be protected in this invention has good sEH inhibitory activity, and some compounds are comparable to or significantly superior to the positive control t-AUCB.

[0190] 2. Determination of the anti-inflammatory effects of compounds in Examples 1, 3, 10, 15, and 16

[0191] Forty-two male Kunming mice (20–25 g) were randomly divided into seven groups: Example 1 group, Example 3 group, Example 10 group, Example 15 group, Example 16 group, celecoxib group, and blank solvent group, with six mice in each group. An inflammatory swelling model was established by subcutaneous injection of 50 μL of 1% λ-carrageenan saline solution into the paw. One hour before modeling, mice were intraperitoneally injected with either a 100 mg / kg solution of the corresponding compound (20 mg / mL, 10% PEG 400 and 10% Tween 80 aqueous solution) or a blank solvent. The initial paw thickness before modeling was taken as 100%, and changes in paw thickness were measured. The degree of paw swelling was statistically analyzed, as shown in Figure 1.

[0192] The results showed that the compounds in Examples 1, 3, 10, 15 and 16 could significantly inhibit carrageenan-induced paw edema in mice, and their effects were superior to celecoxib.

[0193] 3. Determination of the anti-stroke effect of compounds in Examples 1, 3, 10, 15 and 16

[0194] 3.1 Therapeutic effect on acute phase of ischemic stroke

[0195] A middle cerebral artery occlusion (MCAO) reperfusion model was established in male SD rats using the suture occlusion method. After 2 hours of ischemia, the suture was removed to restore perfusion. Immediately after the start of reperfusion, rats were injected via the tail vein with 1 mg / kg or 3 mg / kg of the compound solution described in the above examples (1.5 mg / mL, an aqueous solution of 10% PEG 400 and 10% Tween 80), t-AUCB solution (1.5 mg / mL, an aqueous solution of 10% PEG 400 and 10% Tween 80), edaravone injection (commercially available, 20 mL: 30 mg), or a blank solvent, with 8 rats in each group. After 24 hours, the rats were euthanized under 10% chloral hydrate anesthesia, and the brain tissue was harvested and cut into 4-5 consecutive equidistant coronal sections. The brain sections were completely immersed in 2% TTC (2,3,5-triphenyltetrazolium chloride) solution for staining and photographing, and the infarct volume was calculated, as shown in Figure 2.

[0196] The results showed that the compounds in Examples 1, 3, 10, 15 and 16 significantly reduced the infarct volume in MCAO rats, with better effects than t-AUCB and edaravone.

[0197] Thirty rats were randomly divided into five groups: sham-operated group, solvent group, compound of Example 3 (3 mg / kg) group, compound of Example 16 (3 mg / kg) group, and t-AUCB (3 mg / kg) group. A MCAO reperfusion model was established as described above, and the drug was administered via tail vein immediately after reperfusion. Twenty-four hours later, ischemic brain tissue was harvested and weighed. PBS buffer was added at a weight-to-volume ratio of 1:9, and the tissue was ground at low temperature for 1 min. The tissue was then centrifuged at 3000 rpm for 20 min at 4°C, and the supernatant was collected. The concentrations of TNF-α and IL-1β in the brain tissue were measured using an enzyme-linked immunosorbent assay (ELISA) kit (referring to the ELISA kit instructions), as shown in Figure 3.

[0198] The results showed that the compounds in Examples 3 and 16 could significantly reduce the levels of inflammatory factors in the brain tissue of MCAO rats, with better effects than t-AUCB.

[0199] 3.2 Long-term therapeutic effects on ischemic stroke

[0200] Seventy-eight male SD rats were used to establish a MCAO reperfusion model and were divided into four groups: sham-operated group, solvent group, compound of Example 3 (3 mg / kg) group, compound of Example 16 (3 mg / kg) group, t-AUCB (3 mg / kg) group, and edaravone (3 mg / kg) group. Eight rats were in the sham-operated group, and the remaining groups each had 14 rats. After ischemia-reperfusion, the rats were administered the drug once daily via tail vein injection (experimental drug same as in 3.1) for two consecutive weeks. Animal survival was recorded daily, and the results are shown in Figure 4. Starting in the third week, the Morris water maze test was conducted. Rats were placed in the water facing the pool wall, and the latency to find the platform within 60 seconds was recorded. Rats that found the platform within 60 seconds were guided to the platform and remained there for 30 seconds. The latency to find the platform was recorded over four days to evaluate the rats' spatial memory ability, and the experimental results are shown in Figure 5.

[0201] The results showed that the compounds in Examples 3 and 16 could improve the survival rate of rats, significantly reduce the escape latency of rats, and enhance their learning and memory abilities, with better effects than t-AUCB and edaravone.

[0202] 4. Determination of the hypoglycemic effect of the compounds in Examples 1, 3, 10, 15 and 16

[0203] Forty-eight male BALB / c mice (18–22 g) were randomly divided into eight groups: 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, with six mice in each group. Except for the normal control group, mice were intraperitoneally injected with 1% streptozotocin (STZ) solution (pH 4.5, 0.1 mol / L citrate buffer) once daily for five consecutive days at a dose of 50 mg / kg to establish a diabetic model. Starting from the first day of modeling, mice were intraperitoneally injected once daily with either a 10 mg / kg solution of the corresponding compound (6 mg / mL, 10% PEG 400 and 10% Tween 80 aqueous solution) or a blank solvent. Fasting blood glucose levels were measured after two weeks, as shown in Figure 6.

[0204] The results showed that the blood glucose levels in the treated mice were significantly lower than those in the blank solvent group. The compounds in Examples 1, 3, 10, 15 and 16 could alleviate STZ-induced hyperglycemia, and the compounds in Examples 3 and 16 were more effective than t-AUCB.

[0205] 5. Determination of the antifibrotic effect of the compounds in Examples 1, 3, 10, 15 and 16

[0206] Forty-eight male C57BL6 / J mice (18–22 g) were randomly divided into eight groups: sham-operated group, blank solvent group, Example 1 group, Example 3 group, Example 10 group, Example 15 group, Example 16 group, and t-AUCB group, with six mice in each group. After anesthetizing the mice, the left ureter was ligated and cut to establish a unilateral ureteral obstruction (UUO) model. In the sham-operated group, only the ureter was isolated, without ligation. Starting three days before modeling, mice were intraperitoneally injected with 10 mg / kg of the corresponding compound solution (prepared as in section 4) or blank solvent once daily. Seven days after modeling, the animals were sacrificed, and the kidneys were harvested for Masson staining to examine histopathological changes. The experimental results are shown in Figure 7.

[0207] The results showed that the compounds in Examples 1, 3, 10, 15 and 16 could significantly reduce the fibrotic area in mouse kidney tissue, with the compounds in Examples 3 and 16 being more effective than t-AUCB.

[0208] 6. Determination of the analgesic effect of the compounds in Examples 1, 3, 10, 15 and 16

[0209] 6.1 Determination of analgesic effect

[0210] (1) Male SD rats (200-250g) had their initial mechanical withdrawal reflex threshold (MWT) measured using a Von Frey electronic analgesia meter. The rats were then anesthetized, and the skin was longitudinally incised along the left femur. The muscles were bluntly dissected to expose the sciatic nerve trunk and its distal branches. Surrounding adhesions and the three branches of the sciatic nerve—the tibial, common peroneal, and sural nerves—were separated. The tibial and common peroneal nerves were tightly ligated with silk sutures, while approximately 2-4 mm of the nerve was cut distal to the ligation, preserving and avoiding damage to the sural nerve. This established a selective sciatic nerve branch injury (SNI) pain model. One week after surgery, the MWT was measured to confirm the SNI model.

[0211] After successful model establishment, the SNI model rats were divided into 8 groups: Example 1 group, Example 3 group, Example 10 group, Example 15 group, Example 16 group, EC5026 group, pregabalin group, and blank solvent group, with 6 rats in each group. The rats were administered the 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. With the pre-administration MWT as 100%, the changes in MWT at different time points after administration were measured, and the MWT%-time curve was plotted, and the area under the curve (AUC) was calculated. The experimental results are shown in Figure 8.

[0212] (2) Male SD rats (180-220g) were injected intraperitoneally with 1% streptozotocin (STZ) solution (0.1mol / L citrate buffer with pH 4.5) at a dose of 60mg / kg to establish a diabetic model. One week later, fasting blood glucose was measured to be greater than 11.1mmol / L to confirm the diabetic model. Two weeks later, MWT was measured to confirm the diabetic neuropathic pain (DNP) model.

[0213] DNP model rats were divided into 7 groups: Example 1 group, Example 3 group, Example 10 group, Example 15 group, Example 16 group, pregabalin group, and blank solvent group, with 6 rats in each group. Each group was administered the 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 change in MWT at different time points after administration was measured with the pre-administration MWT as 100%, and the MWT%-time curve was plotted. The area under the curve (AUC) was calculated. The experimental results are shown in Figure 9.

[0214] (3) C57BL6 / J mice (18–22 g) were intraperitoneally injected with paclitaxel solution (0.2 mg / mL, solvent: DMSO-PEG400-Tween 80-physiological saline (1:1:1:7, V / V)) at 1, 3, 5, and 7 days, at a dose of 2 mg / kg. MWT was monitored every 4 days after model initiation using a Von Frey fiber optic pain meter to confirm the paclitaxel-induced neuropathic pain (PINP) model.

[0215] After successful model establishment, the PINP model mice were divided into 7 groups: Example 1 group, Example 3 group, Example 10 group, Example 15 group, Example 16 group, pregabalin group, and blank solvent group, with 6 mice in each group. Each group was administered the 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. With the pre-administration MWT as 100%, the changes in MWT at different time points after administration were measured, and the MWT%-time curve was plotted, and the area under the curve (AUC) was calculated. The experimental results are shown in Figure 10.

[0216] The results showed that the compounds in Examples 1, 3, 10, 15 and 16 could significantly improve the MWT in SNI rats, DNP rats and PINP mice and relieve neuropathic pain. The compounds in Examples 3 and 16 were more effective than EC5026 and pregabalin.

[0217] 6.2 Measurement of inflammatory factors in the spinal cord

[0218] Thirty SD rats (200-250g) were randomly divided into five groups: Example 3 group, Example 16 group, EC5026 group, blank solvent group, and sham-operated group, with six rats in each group. The SNI model was established according to the method described in section 6.1. In the sham-operated group, only the tibial nerve, common peroneal nerve, and sural nerve were isolated. After successful model establishment, the rats were administered the 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. Two hours after administration, the animals were anesthetized, and their hearts were perfused sequentially with approximately 200 mL of physiological saline and approximately 100 mL of 4% paraformaldehyde. The L4-L6 segment of the spinal cord was harvested, weighed, and added to PBS buffer at a weight-to-volume ratio of 1 g:9 mL. The tissue homogenate was then homogenized at 4°C and centrifuged at 2500 rpm for 10 min at 4°C, and the supernatant was collected for later use. The concentrations of inflammatory factors TNF-α and IL-1β in rat spinal cord were determined using an enzyme-linked immunosorbent assay kit (refer to the kit instructions). The results are shown in Figure 11.

[0219] The results showed that the compounds in Examples 3 and 16 could significantly reduce the levels of inflammatory factors in the spinal cord of SNI rats, with better effects than pregabalin.

[0220] 6.3 Effects on motor coordination in rats

[0221] The effects of the compound on motor coordination in rats were evaluated using a rotarod fatigue test. Forty-two male SD rats were randomly divided into seven groups: Example 1 group, Example 3 group, Example 10 group, Example 15 group, Example 16 group, pregabalin group, and a normal control group, with six rats in each group. For the three days prior to the test, rats were trained three times daily at a rotation speed of 10–20 rpm for 5 minutes each time, with a 20-minute interval between training sessions as the fatigue recovery time. On the day of the test, rats were administered the corresponding compound solution (8 mg / mL, 10% PEG 400 and 10% Tween 80 aqueous solution) or a blank solvent via gavage at a dose of 20 mg / kg. The walking time of the rats on the rotarod at a rotation speed of 25 rpm was recorded before and after administration. The walking time of the rats on the rotarod is shown in Figure 12.

[0222] The results showed that the walking time of rats in the pregabalin group was significantly reduced on the rotundus. The compounds in Examples 1, 3, 10, 15 and 16 did not affect the walking time of rats on the rotundus, and there were no adverse central nervous system reactions caused by pregabalin.

[0223] 7. Determination of the antidepressant effects of compounds from Examples 1, 3, 10, 15, and 16

[0224] Forty-two male Kunming mice (20-25g) were randomly divided into seven groups: Example 1 group, Example 3 group, Example 10 group, Example 15 group, Example 16 group, blank solvent group, and normal control group, with six mice in each group. The Kunming mice underwent a 10-minute pre-swimming test. Except for the normal control group, the mice were intraperitoneally injected with 0.5 mg / kg lipopolysaccharide saline solution (0.1 mg / mL). Twenty-three hours later, they were subcutaneously injected with either a 10 mg / kg solution of the corresponding compound (6 mg / mL, 10% PEG 400 and 10% Tween 80 aqueous solution) or a blank solvent. One hour later, a forced swimming test was performed for 6 minutes. The immobility time (s) of each group was recorded within the last 4 minutes. The statistical analysis of the immobility time of the mice during forced swimming is shown in Figure 13.

[0225] The results showed that the compounds in Examples 1, 3, 10, 15 and 16 could significantly reduce the immobility time of mice during forced swimming.

[0226] 8. Determination of the dry eye relief effects of the compounds in Examples 1, 3, 10, 15 and 16

[0227] Seventy-two male BALB / c mice (18–22 g) were randomly divided into eight groups: Example 1, Example 3, Example 10, Example 15, Example 16, cyclosporine eye drops, blank solvent, and a normal control group, with nine mice in each group. Except for the normal control group, mice were treated with 0.2% benzalkonium chloride (BAC) saline solution three times daily for 15 days to establish a dry eye model. Intervention treatment was performed using the compound prepared in the above examples (2 mg / mL, 10% hydroxypropyl-β-cyclodextrin solution), cyclosporine eye drops (II) (commercially available, 0.4 mL: 0.2 mg), or blank solvent, with 5 μL administered to each eye three times daily for 15 days. Tear secretion and corneal fluorescein staining scores were measured at different time points, as shown in Figure 14.

[0228] The results showed that the compounds in Examples 1, 3, 10, 15 and 16 could significantly alleviate BAC-induced dry eye, increase tear secretion in mice, and alleviate corneal damage, with better effects than cyclosporine eye drops (II).

[0229] Seventy-five male BALB / c mice (18–22 g) were randomly divided into five groups: Example 3 group, Example 16 group, cyclosporine eye drops (II) group, blank solvent group, and normal control group, with 15 mice in each group. A dry eye model and drug administration were performed as described above. Fifteen days later, the mice were sacrificed, and corneas were collected. Six corneas from every three mice were combined into one sample, and three samples were collected from each group for determining the levels of 14,15-EET and 14,15-DHET in the tissue. The remaining corneas were combined into one sample from every four mice, and three samples were collected from each group for determining the concentration of TNF-α in the tissue. The levels of 14,15-EET and 14,15-DHET were determined using LC-MS / MS, and the concentration of TNF-α was determined using an enzyme-linked immunosorbent assay (ELISA) kit, following the kit's instructions. Figure 15 shows the levels of 14,15-EET and 14,15-DHET in the tissue, and Figure 16 shows the concentration of the inflammatory factor TNF-α in the tissue.

[0230] The results showed that the compounds in Examples 3 and 16 could increase the level of 14,15-EET in the cornea and reduce the content of 14,15-DHET and TNF-α, and their effects were better than those of cyclosporine eye drops (II).

[0231] 9. Determination of the therapeutic effects of the compounds in Examples 3 and 16 on diabetic retinopathy

[0232] Eighty male C57BL / 6J mice (18–22 g) were randomly divided into five groups: Example 3 group, Example 16 group, t-AUCB group, blank solvent group, and normal control group, with 16 mice in each group. Except for the normal control group, mice were intraperitoneally injected with 1% streptozotocin (STZ) solution (0.1 mol / L citrate buffer, pH 4.5) at a dose of 50 mg / kg, once daily for 5 consecutive days. One week later, a random blood glucose level ≥16.7 mmol / L was considered a successful establishment of a diabetic model, and intervention treatment was initiated using eye drops containing the compounds from Example 3 and Example 16 (1 mg / mL, 10% hydroxypropyl-β-cyclodextrin solution) or the blank solvent, administered twice daily, 5 μL per eye each time. Diabetic mice were fed for 6 months to induce retinopathy. After 6 months, the mice were injected intravenously with 0.1 mL of 2% Evans blue saline solution for 2 hours. After cardiac perfusion with 50 mL of saline, retinal tissue was harvested, homogenized, centrifuged, and the supernatant was collected. The absorbance was measured at 620 nm to examine retinal vascular permeability. Separately, retinal tissue was treated with 5% pepsin solution and pepsin to remove excess tissue, obtaining retinal vessels. After PAS staining, microscopic imaging was performed to observe the morphology of retinal vessels and pericytes, and the number of free pericytes in the retina was counted. The experimental results are shown in Figures 17 and 18.

[0233] The results showed that the compounds in Examples 3 and 16 could significantly reduce the permeability of retinal blood vessels in diabetic mice, reduce the number of free pericytes and cell-free vessels in retinal capillaries, maintain vascular homeostasis, and alleviate the symptoms of diabetic retinopathy, with better effects than t-AUCB.

[0234] The distribution of the compounds from Examples 3 and 16 in ocular tissues was studied using Dutch rabbits. Six male Dutch rabbits (1.6–2.4 kg) were divided into two groups of three each. Each group received 50 μL of eye drops containing the compounds from Examples 3 and 16, respectively. The animals were sacrificed 0.5 h after administration, and samples of cornea, aqueous humor, bulbar conjunctiva, iris, vitreous body, choroid, retina, and sclera were immediately collected. The samples were homogenized with 20% methanol at a weight-to-volume ratio of 1:10, and the compound content in the supernatant was determined by LC-MS / MS. The experimental results are shown in Figure 19.

[0235] The results showed that the compounds in Examples 3 and 16 could reach the posterior retinal tissue after being administered as eye drops.

[0236] 10. Effects of the compounds in Examples 3 and 16 on the levels of 14,15-EET and 14,15-DHET in rats.

[0237] Male SD rats (250–300 g) were administered the test compounds of Examples 3 and 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. Approximately 0.25 mL of blood was collected from the jugular sinus before administration and at 15 min, 30 min, 1 h, 2 h, 4 h, and 8 h after administration, and placed in EDTA-K2 anticoagulant tubes. The whole blood samples were centrifuged at 1500–1600 rpm for 10 min, and the separated plasma was stored at -40–-20°C. The concentrations of 14,15-EET and 14,15-DHET in rat plasma were determined using liquid chromatography-mass spectrometry (LC-MS / MS). The results are shown in Figure 20.

[0238] The results showed that the compounds in Example 3 and Example 16, as well as EC5026, could significantly increase the level of 14,15-EET and decrease the level of 14,15-DHET in rat plasma, with better effects than EC5026.

[0239] 11. Molecular docking results of EC5026, compounds from Examples 1, 3, and 16 with sEH

[0240] Using sEH cocrystal protein (PDB code: 4OCZ) as the docking model, The 4.5.208 software was used to perform molecular docking of EC5026, compounds from Examples 1, 3, and 16 with sEH, respectively.

[0241] The molecular docking results of EC5026 and sEH are shown in Figure 21. The 3-fluoro-4-(trifluoromethoxy)phenyl group in the EC5026 structure faces the hydrophobic region inside the cavity of the sEH protein. The phenyl group forms a π-π stacking interaction with the His524 residue, and the fluorine atom at position 3 forms a halogen bond with the Phe267 residue. The two NH groups of the urea fragment form a bidentate hydrogen bond with the Asp335 residue, and the carbonyl oxygen atom forms a hydrogen bond with the Tyr383 and Tyr466 residues, respectively. The (S)-2-methylbutyramide fragment extends into the solvent region.

[0242] The molecular docking results of the compound in Example 1 with sEH are shown in Figure 22. The 4-chlorophenyl group in the compound forms a π-π stacking interaction with the Phe267 residue in the hydrophobic region inside the cavity of the sEH protein; 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 with Gln384; the two NH atoms of the urea fragment form a bidentate hydrogen bond with the Asp335 residue, and the oxygen atom of the urea fragment forms a hydrogen bond with the Tyr383 and Tyr466 residues, respectively; the (4-morpholino)ethyl moiety faces the solvent region.

[0243] The molecular docking results of the compound in Example 3 with sEH are shown in Figure 23. The 4-chlorophenyl group in the compound forms a π-π stacking interaction with the Phe267 residue in the hydrophobic region inside the cavity of the sEH protein; the benzothiazole structure forms an additional π-π stacking interaction with the Trp336 residue; the two NH groups of the urea fragment form a bidentate hydrogen bond with the Asp335 residue, and the oxygen atoms of the urea fragment form hydrogen bonds with the Tyr383 and Tyr466 residues, respectively; the (4-morpholino)ethyl moiety faces the solvent region.

[0244] The molecular docking results of the compound in Example 16 with sEH are shown in Figure 24. The 4-chlorophenyl group in the compound forms a π-π stacking interaction with the Phe267 residue in the hydrophobic region inside the cavity of the sEH protein; 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 NH atoms of the urea fragment form a bidentate hydrogen bond interaction with the Asp335 residue, and the oxygen atom of the urea fragment forms hydrogen bonds with the Tyr383 and Tyr466 residues, respectively; the (2-oxo-1-pyrrolidinyl)ethyl moiety faces the solvent region, and the ketone carbonyl group forms an additional hydrogen bond interaction with the Ile363 residue.

[0245] Compared to EC5026, the compounds in Examples 1, 3, and 16 not only maintained the key interaction forces equivalent to those between EC5026 and the sEH protein, but their benzothiazole structure also generated additional interaction forces with sEH, enabling them to bind better to the sEH protein and thus exhibiting superior inhibitory activity against sEH.

[0246] In summary, in the compounds of general formula I of this invention, the substituent R... 1 R 2 R 3 R 4 Structural modifications such as m, n, X, and Y are made to enable better binding with sEH, resulting in a good inhibitory effect on sEH and good therapeutic effects on inflammation, stroke, diabetes, fibrosis, pain, depression, and dry eye syndrome. This demonstrates that the interaction of specific substituents with sEH can achieve unexpected effects.

Claims

1. A 2-substituted aminobenzothiazole compound, characterized in that: 2-Substituted aminobenzothiazole compounds are compounds represented by general formula I and their pharmaceutically acceptable salts. in, When Y represents N atom, X represents S atom; when Y represents S atom, X represents N atom. R 1 and R 2 Each is independently selected from hydrogen, (C1-C6)alkyl, (C3-C7)cycloalkyl, or R 1 and R 2 Together with the nitrogen atoms they are attached to, they form 4-10 membered heterocyclic groups, wherein the heterocyclic group, in addition to the nitrogen atoms, optionally contains 0-3 heteroatoms selected from N, O, or S, and the heterocyclic group may optionally be coupled with 1-3 identical or different R atoms. 5 The heterocyclic group is substituted with 0-2 carbon-carbon double bonds; R 5 It can be a hydrogen atom, =O, hydroxyl, amino, halogen, carboxyl, cyano, nitro, (C1-C6)alkyl, (C1-C6)alkoxy, (C3-C7)cycloalkyl, (C1-C6)alkylsulfinyl, (C1-C6)alkylsulfonyl or (C1-C6)alkylformyl. R 3 For (C6-C) 10 ) aryl, 5-10 heteroaryl or adamantyl, wherein the heteroaryl contains 1-3 heteroatoms selected from N, O or S, and the aryl or heteroaryl may optionally be surrounded by 1-3 identical or different R atoms. 6 replace; R 6 The following groups are represented by hydrogen atom, hydroxyl group, halogen, cyano group, mercapto group, carboxyl group, carbamoyl group, substituted or unsubstituted (C1-C6) alkyl group, substituted or unsubstituted (C1-C6) alkoxy group, substituted or unsubstituted (C1-C6) alkylthio group, substituted or unsubstituted (C2-C6) alkenyl group, substituted or unsubstituted (C1-C6) alkylsulfinyl group, substituted or unsubstituted (C1-C6) alkylsulfonyl group, substituted or unsubstituted (C1-C6) alkylformyl group, substituted or unsubstituted (C1-C6) alkoxyformyl group, -CO-NHR 7 -CO-NR 7 R 8 -NH-CO-R 7 -NHR 7 or -NR 7 R 8 Among them, R 7 R 8 The substituted or unsubstituted (C1-C6) alkyl groups are the same or different; the substituents of each of the above substitutions are selected from hydroxyl, amino or halogen; the substitution is monosubstituted or polysubstituted, and the substituents of the polysubstituted groups are the same or different; R 4 It is hydrogen, halogen, (C1-C4)alkyl, or (C1-C4)alkoxy; m is 0, 1, or 2; n is 2, 3, or 4.

2. The 2-substituted aminobenzothiazole compound according to claim 1, characterized in that: In the compound, When Y represents N atom, X represents S atom; when Y represents S atom, X represents N atom. R 1 and R 2 Together with the nitrogen atoms they are attached to, they form 4-10 membered heterocyclic groups, wherein the heterocyclic group, in addition to the nitrogen atoms, optionally contains 0-3 heteroatoms selected from N, O, or S, and the heterocyclic group may optionally be bonded by 1-3 identical or different R atoms. 5 The heterocyclic group is substituted with 0-2 carbon-carbon double bonds; R 5 It can be a hydrogen atom, =O, hydroxyl, amino, halogen, carboxyl, cyano, nitro, (C1-C4)alkyl, (C1-C4)alkoxy, (C3-C6)cycloalkyl, (C1-C4)alkylsulfinyl, (C1-C4)alkylsulfonyl or (C1-C4)alkylformyl. R 3 It is a phenyl, a 5-6-membered heteroaryl, or an adamantyl group, wherein the heteroaryl group contains 1-3 heteroatoms selected from N, O, or S, and the phenyl or the heteroaryl group may optionally be surrounded by 1-3 identical or different R atoms. 6 replace; R 6 The following groups are represented by hydrogen atom, hydroxyl group, halogen, cyano group, mercapto group, carboxyl group, carbamoyl group, substituted or unsubstituted (C1-C4) alkyl group, substituted or unsubstituted (C1-C4) alkoxy group, substituted or unsubstituted (C1-C4) alkylthio group, substituted or unsubstituted (C2-C4) alkenyl group, substituted or unsubstituted (C1-C4) alkylsulfinyl group, substituted or unsubstituted (C1-C4) alkylsulfonyl group, substituted or unsubstituted (C1-C4) alkylformyl group, substituted or unsubstituted (C1-C4) alkoxyformyl group, -CO-NHR 7 -CO-NR 7 R 8 -NH-CO-R 7 -NHR 7 or -NR 7 R 8 Among them, R 7 R 8 The substituted or unsubstituted (C1-C4) alkyl groups are the same or different; the substituents of each of the above substitutions are selected from hydroxyl, amino or halogen; the substitution is monosubstituted or polysubstituted, and the substituents of the polysubstituted groups are the same or different; R 4 It can be hydrogen, halogen, or methyl; m is 0 or 1; n is 2 or 3.

3. The 2-substituted aminobenzothiazole compound according to claim 2, characterized in that: The compound When Y represents N atom, X represents S atom; when Y represents S atom, X represents N atom. R 1 and R 2 Together with the nitrogen atoms they are attached to, they form R 3 It is phenyl or adamantyl; the phenyl group is optionally surrounded by 1-3 identical or different R groups. 6 replace; R 6 Halogen, unsubstituted or optionally substituted with 1-3 identical or different halogens (C1-C4) alkyl or (C1-C4) alkoxy; R 4 It can be hydrogen, fluorine, or chlorine; m is 0 or 1; n is 2.

4. The 2-substituted aminobenzothiazole compound according to claim 3, characterized in that: The compound When Y represents N atoms, X represents S atoms; When Y represents an S atom, X represents an N atom, and R 4 For H; R 1 and R 2 Together with the nitrogen atoms they are attached to, they form R 3 It is a phenyl group, wherein the para position of the phenyl group is R 6 replace; R 6 It is a halogen, trifluoromethyl or trifluoromethoxy; R 4 It can be hydrogen, fluorine, or chlorine; m is 0 or 1; n is 2.

5. The 2-substituted aminobenzothiazole compound according to claim 4, characterized in that: In the compound, when Y represents an N atom, X represents an S atom, which is shown in general formula (II). R 1 and R 2 Together with the nitrogen atoms they are attached to, they form R 3 It is a phenyl group, wherein the para position of the phenyl group is R 6 replace; R 6 It is a halogen, trifluoromethyl or trifluoromethoxy; R 4 It can be hydrogen, fluorine, or chlorine; m is 0 or 1; n is 2.

6. The 2-substituted aminobenzothiazole compound according to claim 4, characterized in that: The compound is 1-{N-[2-(4-morpholinyl)ethyl]-2-aminobenzo[d]thiazolyl-5-yl}-3-(4-chlorophenyl)urea; 1-{N-[2-(2-oxo-1-pyrrolidinyl)ethyl]-2-aminobenzo[d]thiazolyl-5-yl}-3-(4-chlorophenyl)urea; 1-{N-[2-(4-morpholinyl)ethyl]-2-aminobenzo[d]thiazolyl-6-yl}-3-(4-chlorophenyl)urea; 1-{N-[2-(dimethylamino)ethyl]-2-aminobenzo[d]thiazolyl-6-yl}-3-(4-chlorophenyl)urea; 1-{N-[2-(4-morpholinyl)ethyl]-2-aminobenzo[d]thiazolyl-5-yl}-3-(3-fluoro-4-chlorophenyl)urea; 1-{N-[2-(1-piperidinyl)ethyl]-2-aminobenzo[d]thiazolyl-5-yl}-3-(4-chlorophenyl)urea; 1-{N-[2-(4-morpholinyl)ethyl]-2-aminobenzo[d]thiazolyl-6-yl}-3-(4-trifluoromethoxyphenyl)urea; 1-{N-[2-(2-oxo-1-pyrrolidinyl)ethyl]-2-aminobenzo[d]thiazolyl-5-yl}-3-(3-fluoro-4-chlorophenyl)urea; 1-{N-[2-(4-morpholinyl)ethyl]-2-aminobenzo[d]thiazolyl-6-yl}-3-(1-adamantyl)urea; 1-{N-[2-(2-oxo-1-pyrrolidinyl)ethyl]-2-aminobenzo[d]thiazolyl}-3-(4-chlorophenyl)urea; 1-{N-[2-(3-oxo-4-morpholinyl)ethyl]-2-aminobenzo[d]thiazolyl-5-yl}-3-(3,4-dichlorophenyl)urea; 1-{N-[2-(3-oxo-4-morpholinyl)ethyl]-2-amino-5-fluorobenzo[d]thiazolyl-6-yl}-3-(4-chlorophenyl)urea; 1-{N-[2-(2-oxo-1-piperidinyl)ethyl]-2-amino-5-fluorobenzo[d]thiazolyl-6-yl}-3-(4-chlorophenyl)urea; 1-{N-[2-(3-oxo-4-morpholinyl)ethyl]-2-aminobenzo[d]thiazo-6-yl}-3-(1-adamantyl)urea; 1-{N-[2-(3-oxo-4-morpholinyl)ethyl]-2-amino-5-chlorobenzo[d]thiazolyl-6-yl}-3-(4-chlorophenyl)urea; 1-{N-[2-(2-oxo-1-pyrrolidinyl)ethyl]-2-amino-5-fluorobenzo[d]thiazolyl-6-yl}-3-(4-chlorophenyl)urea.

7. The 2-substituted aminobenzothiazole compound according to any one of claims 1 to 6, characterized in that: The pharmaceutically acceptable salt of the 2-substituted aminobenzothiazole compound is a salt formed by the 2-substituted aminobenzothiazole compound and 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.

8. A pharmaceutical composition, characterized in that: The pharmaceutical composition contains one or more of the 2-substituted aminobenzothiazole compounds according to any one of claims 1 to 7 and their pharmaceutically acceptable salts, as well as a pharmaceutically acceptable carrier.

9. The use of a 2-substituted aminobenzothiazole compound according to any one of claims 1 to 7, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 8 in the preparation of a medicament for treating and / or preventing sEH-mediated diseases.

10. The application according to claim 9, characterized in that: The diseases mentioned include inflammatory diseases, cardiovascular and cerebrovascular diseases, diabetes, diabetes complications, diabetes-related diseases, fibrotic diseases, neurological and psychiatric diseases, pain, ulcerative diseases, and dry eye syndrome.

11. The application according to claim 10, characterized in that, The inflammatory diseases include inflammatory liver disease, inflammatory kidney disease, inflammatory lung disease, inflammatory brain disease, myocarditis, pancreatitis, arthritis, soft tissue inflammation, osteomyelitis, 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 1 diabetes or type 2 diabetes; the complications of diabetes include diabetic retinopathy, diabetic uveitis, diabetic cataracts, diabetic nephropathy, diabetic skin diseases, and diabetic peripheral neuropathy; the diabetes-related diseases include hyperlipidemia, hyperuricemia, and hyperuricemia. Acidosis and gout, obesity and metabolic syndrome; the fibrotic diseases include pulmonary fibrosis, liver fibrosis, myocardial fibrosis and renal fibrosis; the neurological and psychiatric diseases include Alzheimer's disease, epilepsy, Parkinson's disease, amyotrophic lateral sclerosis, schizophrenia, mental disorders, depression and neurasthenia; the pain diseases include neuropathic pain, inflammatory pain, tumor pain and mixed pain; the ulcerative diseases include gastric ulcer, duodenal ulcer, ulcerative colitis, corneal ulcer and oral ulcer; the dry eye syndrome includes aqueous hypoplasia dry eye, lipid abnormality dry eye, mucin abnormality dry eye, tear dynamics abnormality dry eye and mixed dry eye.

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