2-substituted aminobenzothiazole compound and application thereof

Abstract

The invention relates to a 2-substituted aminobenzothiazole compound and a pharmaceutically acceptable salt thereof, as well as a pharmaceutical composition containing the compound and application thereof. The 2-substituted aminobenzothiazole compound is a compound as shown in a general formula I and pharmaceutically acceptable salts thereof, wherein substituent groups R1, R2, R3, R4, m, n, X and Y have meanings given in the specification. The invention also relates to application of the compound shown in the general formula I in preparation of drugs for treating and / or preventing sEH-mediated diseases, in particular to application of the compound shown in the general formula I in preparation of drugs for treating inflammatory diseases, cardiovascular and cerebrovascular diseases, pain, xerophthalmia, diabetes, diabetic complications, diabetes-related diseases, fibrosis diseases, neurological and mental diseases, ulcerative diseases and the like. # imgabs0 #

Description

Technical Field

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

[0002] Epoxide hydrolases (EH) are commonly found in mammals, plants and microorganisms in nature, and can efficiently catalyze the hydrolysis of epoxides to generate vicinal diols. Eight subtypes of epoxide hydrolases have been discovered, including 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 more deeply in recent years. 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 the EPHX2 gene, which is located in the P21-P12 region of human chromosome 8. It includes 18 introns and 19 exons, encoding more than 550 amino acids and a total length of 45kb. sEH is composed of two 60kD subunits in an antiparallel manner, each of which contains two different enzymatic activity domains. The C-terminus has an α / β hydrolase folding structure, which can act as a catalytic subunit to hydrolyze epoxides to form corresponding diols; the N-terminus has phosphatase activity and can hydrolyze a variety of lipid phosphates. sEH is widely distributed in many organs of the human body, such as the brain, liver, and kidney, while its content is relatively low 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, lipoxins and leukotrienes. AA can be converted into epoxyeicosatrienoic acids (EETs) with high biological activity by cytochrome P450 (CYP450). EETs have anti-inflammatory, anti-apoptotic and antioxidant effects and participate in the pathophysiological processes of various diseases such as the immune system, nervous system and metabolic system. sEH is the main metabolic enzyme of EETs and can quickly hydrolyze EETs into dihydroxyeicosatrienoic acids (DHETs) with low biological activity, affecting the protective effect of EETs. sEH inhibitors can achieve the purpose of treating various diseases by increasing the level of endogenous EETs and maintaining the biological activity of EETs.

[0005] Currently, no sEH inhibitors have been used in clinical practice. The sEH inhibitors reported in the literature include t-AUCB, AR9281, GSK2256294 and EC5026. GSK2256294 is a small molecule sEH inhibitor discovered by GlaxoSmithKline, which can be used to treat diseases such as diabetes and obesity and is currently in Phase II clinical trials. EC5026 is a urea sEH inhibitor discovered by Bruce Hammock's team, and the compound is currently in Phase I clinical trials.

[0006] Summary of the invention

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

[0008] To achieve the above purpose, the present invention adopts the following technical solution:

[0009] The present invention relates to 2-substituted aminobenzothiazole compounds and pharmaceutically acceptable salts thereof as shown in general formula I.

[0010]

[0011] in,

[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, 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, and which may be optionally replaced by 1-3 identical or different R 5 Substituted, heterocyclic group contains 0-2 carbon-carbon double bonds;

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

[0015] R 3 is (C6-C 10 ) aryl, 5-10 membered heteroaryl or adamantyl, the heteroaryl containing 1-3 heteroatoms selected from N, O or S, and the aryl or heteroaryl may be optionally replaced by 1-3 identical or different R 6 replace;

[0016] R 6 is a hydrogen atom, a hydroxyl group, a halogen group, a cyano group, a mercapto group, a carboxyl group, a carbamoyl group, a substituted or unsubstituted (C1-C6) alkyl group, a substituted or unsubstituted (C1-C6) alkoxy group, a substituted or unsubstituted (C1-C6) alkylthio group, a substituted or unsubstituted (C2-C6) alkenyl group, a substituted or unsubstituted (C1-C6) alkylsulfinyl group, a substituted or unsubstituted (C1-C6) alkylsulfonyl group, a substituted or unsubstituted (C1-C6) alkylformyl group, a substituted or unsubstituted (C1-C6) alkoxyformyl group, or -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 are the same or different substituted or unsubstituted (C1-C6) alkyl groups; the substituents of the above-mentioned substitutions are selected from hydroxyl, amino or halogen; the substitutions are mono- or poly-substituted, and the substituents of the poly-substituted groups 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 invention preferably relates to 2-substituted aminobenzothiazole compounds of general formula I and pharmaceutically acceptable salts thereof,

[0021] in,

[0022] When Y represents an N atom, X represents an S atom; when Y represents an S atom, X represents an N atom;

[0023] R 1 and 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, and the heterocyclic group may also be optionally replaced by 1-3 identical or different R 5 Substituted, heterocyclic group contains 0-2 carbon-carbon double bonds;

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

[0025] R 3 is phenyl, 5-6 membered heteroaryl or adamantyl, the heteroaryl contains 1-3 heteroatoms selected from N, O or S, and the phenyl or heteroaryl may be optionally substituted by 1-3 identical or different R 6 replace;

[0026] R 6 is a hydrogen atom, a hydroxyl group, a halogen group, a cyano group, a mercapto group, a carboxyl group, a carbamoyl group, a substituted or unsubstituted (C1-C4) alkyl group, a substituted or unsubstituted (C1-C4) alkoxy group, a substituted or unsubstituted (C1-C4) alkylthio group, a substituted or unsubstituted (C2-C4) alkenyl group, a substituted or unsubstituted (C1-C4) alkylsulfinyl group, a substituted or unsubstituted (C1-C4) alkylsulfonyl group, a substituted or unsubstituted (C1-C4) alkylformyl group, a substituted or unsubstituted (C1-C4) alkoxyformyl group, or -CO-NHR. 7 、-CO-NR 7 R 8 、-NH-CO-R 7 、-NHR 7 or -NR 7 R 8 ; Among them, R 7 , R 8are the same or different substituted or unsubstituted (C1-C4) alkyl groups; the substituents of the above-mentioned substitutions are selected from hydroxyl, amino or halogen; the substitutions are mono- or poly-substituted, and the poly-substituted substituents are the same 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 invention more preferably relates to 2-substituted aminobenzothiazole compounds of general formula I and pharmaceutically acceptable salts thereof, wherein:

[0031] When Y represents an N atom, X represents an S atom; when Y represents an S atom, X represents an N atom;

[0032] R 1 and R 2 Together with the nitrogen atoms to which they are attached, they form

[0033]

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

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

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

[0037] m is 0 or 1;

[0038] n is 2.

[0039] The present invention particularly preferably relates to 2-substituted aminobenzothiazole compounds of general formula I and pharmaceutically acceptable salts thereof, wherein:

[0040] When Y represents an N atom, X represents an S atom; when Y represents an S atom, X represents an N atom, and R 4 H; R 1 and R 2 Together with the nitrogen atoms to which they are attached, they form

[0041]

[0042] R 3 is phenyl, the para position of phenyl is replaced by R 6 replace;

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

[0048]

[0049] R 1 and R 2 Together with the nitrogen atoms to which they are attached, they form

[0050]

[0051] R 3 is a phenyl group, wherein the para position of the phenyl group is replaced by R 6 replace;

[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 invention provides the following 2-substituted aminobenzothiazole compounds and pharmaceutically acceptable salts thereof:

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

[0073] Optionally, the pharmaceutically acceptable salt of the 2-substituted aminobenzothiazole compound is a salt of the 2-substituted aminobenzothiazole compound and an acid, wherein the acid is 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.

[0074] The present invention also includes prodrugs of the compounds of the present invention. Prodrugs of the compounds of the present invention are compounds of general formula I, which may themselves have weak activity or even no activity, but after administration, are converted into the corresponding biologically active form under physiological conditions (e.g., by metabolism, solvolysis or other means).

[0075] Unless otherwise indicated, the term "halogen" as used herein 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.

[0076] "C1-C4" means that the number of carbon atoms in the defined group (such as alkyl, alkoxy, cycloalkyl, etc.) is 1, 2, 3 or 4. From this, the meaning of other terms described in a similar manner can be inferred, such as "C1-C6", "C3-C6", etc.

[0077] "Alkyl" refers to a straight or branched alkyl group. For example, "C1-C6 alkyl" refers to a straight or branched alkyl group having 1 to 6 carbon atoms, for example, selected from methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, pentyl and hexyl. "Alkenyl" refers to a straight or branched alkenyl group.

[0078] "Cycloalkyl" refers to a substituted or unsubstituted cycloalkyl group. For example, "C3-C6 cycloalkyl" refers to a cycloalkyl group having 3 to 6 carbon atoms, such as cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl.

[0079] "Alkoxy" is a -OR group, "alkylthio" is a -SR group, "alkylsulfinyl" is a -SO-R group, "alkylsulfonyl" is a -SO2-R group, "alkylformyl" is a -CO-R group, and "alkoxyformyl" is a -CO-OR group, where R is alkyl as defined above.

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

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

[0082] The carriers used in the pharmaceutical composition of the present invention are common types available in the pharmaceutical field, including: binders, lubricants, disintegrants, solubilizers, diluents, stabilizers, suspending agents, pigments, flavoring agents, etc. for oral preparations; pH regulators, osmotic pressure regulators, solubilizers, stabilizers, etc. for injectable preparations; bases, diluents, lubricants, preservatives, etc. for local preparations. Pharmaceutical preparations can be administered orally, parenterally (e.g., intravenously, subcutaneously, intraperitoneally, etc.) or topically (e.g., ocular, nasal, sublingual, skin, etc.). If some drugs are unstable under gastric conditions, they can be formulated into enteric-coated tablets.

[0083] Through in vitro and in vivo inhibitory activity tests, it was found that the compounds of general formula I in the present invention and their pharmaceutically acceptable salts have significant inhibitory activity against sEH and can stabilize the level of EETs in the body. The results of pharmacodynamic studies show that the compounds in the present invention can inhibit carrageenan-induced foot swelling, protect ischemic brain damage, maintain blood sugar homeostasis, relieve neuropathic pain, resist tissue fibrosis, relieve depression, improve dry eye symptoms, etc., and can be used to treat sEH-mediated inflammatory diseases, cardiovascular and cerebrovascular diseases, diabetes and complications, fibrotic diseases, neurological and mental diseases, pain, ulcerative diseases or dry eye, etc.

[0084] The exact amount of a compound of the invention required to treat a condition mediated by sEH will vary from subject to subject, depending on the species, age and general condition of the subject, the severity of the disease being treated, the specific compound used and the mode of administration, such as route and frequency of administration, etc. An appropriate effective amount can be determined by one of ordinary skill in the art using only routine experimentation.

[0085] The dosage of the compound can be 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 can vary depending on the patient's needs, the severity of the inflammatory diseases, cardiovascular and cerebrovascular diseases, diabetes, diabetic complications, diabetes-related diseases, fibrotic diseases, neurological and psychiatric diseases, pain, ulcerative diseases or dry eye treated, and the specific compound used. Moreover, it is understood that the initial dose administered can be increased beyond the upper limit in order to quickly reach the desired blood level, or the initial dose can be less than the optimal value, and the daily dose can be gradually increased during treatment, depending on the specific situation. If necessary, the daily dose can also be divided into multiple administrations, such as 2-4 times a day.

[0086] Mammal means a human or an animal.

[0087] The amount of active ingredient, i.e. the compound according to the invention, in the pharmaceutical composition and its unit dosage form may vary, depending on the specific application, the efficacy of the specific compound and the desired concentration. Generally speaking, the content of the active ingredient will be between 0.5% and 90%, based on the total weight of the composition.

[0088] In combination therapy, the compound of the present invention and the other compound may be administered simultaneously or intermittently. When administered simultaneously, the compound of the present invention and the other compound may be combined in a single pharmaceutical composition or in separate compositions.

[0089] The present invention also provides a use of a compound of formula I and a pharmaceutically acceptable salt or pharmaceutical composition thereof in the preparation of a drug for treating and / or preventing sEH-mediated diseases. The 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.

[0090] Specifically, inflammatory diseases include inflammatory liver disease, inflammatory kidney disease, inflammatory lung disease, inflammatory brain disease, myocarditis, pancreatitis, arthritis, soft tissue inflammation, bone tissue inflammation 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 diabetes or type 2 diabetes; diabetic complications include diabetic retinopathy, diabetic uveitis, diabetic cataract, diabetic nephropathy, diabetic skin disease and diabetic peripheral neuropathy; diabetes-related diseases include hyperlipidemia, hyperlipidemia, Uricemia and gout, obesity and metabolic syndrome; fibrotic diseases include pulmonary fibrosis, liver fibrosis, myocardial fibrosis and renal fibrosis; neurological and psychiatric diseases include Alzheimer's disease, epilepsy, Parkinson's disease, amyotrophic lateral sclerosis, schizophrenia, mental disorders, depression, neurasthenia; pain diseases include neuropathic pain, inflammatory pain, tumor pain and mixed pain; ulcerative diseases include gastric ulcer, duodenal ulcer, ulcerative colitis, corneal ulcer and oral ulcer; dry eye includes aqueous deficiency dry eye, lipid abnormality dry eye, mucin abnormality dry eye, tear dynamics abnormality dry eye and mixed dry eye.

[0091] The present invention relates to a method for treating and / or preventing sEH-mediated diseases comprising administering to a subject in need thereof a compound of formula I and a pharmaceutically acceptable salt thereof. The compound of formula I and a pharmaceutically acceptable salt thereof of the present invention exhibit sEH inhibitory activity and can therefore be effectively used to prevent or treat sEH-mediated related diseases.

[0092] In the present 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 administration of the compounds or compositions of the present invention, and the term "treatment" refers to all effects of alleviating or beneficially changing the symptoms of the above-mentioned diseases by administering the compounds or compositions of the present invention.

[0093] The examples and preparations provided below further illustrate and illustrate the compounds of the present invention and their preparation methods. It should be understood that the scope of the following examples and preparations does not limit the scope of the present invention in any way.

[0094] The following synthetic routes describe the preparation of compounds of formula I of the present invention, and all raw materials are prepared by the methods described in these synthetic routes, by methods well known to those of ordinary skill in the field 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 methods similar thereto, which are well known to those of ordinary skill in the field of organic chemistry. All variable factors used in these synthetic routes are as defined below or as defined in the claims.

[0095] The synthetic route of the compound of general formula I is: raw material A reacts with Boc2(O) to obtain intermediate B, intermediate B reacts with R 1 R 2 N(CH2) n Cl undergoes nucleophilic substitution reaction to obtain intermediate C, which is then reduced by ferric acid or H2 / Pd-C to obtain intermediate D; R 3 (CH2) m NH2 reacts with phenyl chloroformate to obtain intermediate E; intermediate D and intermediate E undergo condensation reaction to obtain intermediate F, intermediate F is subjected to the action of hydrogen chloride to remove the Boc protecting group to obtain the hydrochloride of the compound of general formula I, and the latter is then reacted with sodium hydroxide solution to obtain the free compound of general formula I.

[0096] BRIEF DESCRIPTION OF THE DRAWINGS

[0097] Figure 1 This is the anti-inflammatory effect diagram of the compounds 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;

[0098] Figure 2 This is a graph showing the effect of the compounds of the present invention on reducing the volume of cerebral infarction in rats with cerebral ischemia, * compared with the solvent group p<0.05, # compared with the edaravone group p<0.05, $ compared with the t-AUCB group p<0.05;

[0099] Figure 3 This is a graph showing the effect of the compounds of the present invention on reducing the level of inflammatory factors in the brain tissue of rats with cerebral ischemia, * compared with the solvent group p<0.05, # compared with the sham operation group p<0.05, $ compared with the t-AUCB group p<0.05;

[0100] Figure 4 This is a diagram showing the effect of the compounds of the present invention on improving the survival rate of rats with cerebral ischemia.

[0101] Figure 5 The effect diagram of the compounds 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 operation group p<0.05, $ compared with the t-AUCB group p<0.05, & compared with the edaravone group p<0.05;

[0102] Figure 6 The graph is a hypoglycemic effect diagram of the compounds of the present invention, * compared with the solvent group p<0.05, # compared with the normal group p<0.05, $ compared with the t-AUCB group p<0.05;

[0103] Figure 7This is the anti-fibrosis effect diagram of the compounds of the present invention, * compared with the solvent group p < 0.05, # compared with the sham operation group p < 0.05, $ compared with the t-AUCB group p < 0.05;

[0104] Figure 8 This is a graph showing the analgesic effect of the compounds of the present invention on selective injury of sciatic nerve branches, * compared with the solvent group p<0.05, # compared with the EC5026 group p<0.05;

[0105] Fig. 9 This is a graph showing the analgesic effect of the compounds of the present invention on diabetic pathological neuropathy, * compared with the solvent group, p<0.05, # compared with the pregabalin group, p<0.05;

[0106] Fig.10 This is a graph showing the analgesic effect of the compounds of the present invention on paclitaxel-induced neuropathic pain, * compared with the solvent group, p<0.05, # compared with the pregabalin group, p<0.05;

[0107] Fig.11 This is a graph showing the effect of the compounds of the present invention on reducing the level of inflammatory factors in the spinal cord tissue of animals with neuropathic pain, * compared with the solvent group p<0.05, ## compared with the sham operation group p<0.01, $ compared with the EC5026 group p<0.05;

[0108] Fig.12 The effect of the compounds of the present invention on the motor coordination function of rats, **p<0.01 compared with before administration;

[0109] Fig.13 This is the antidepressant effect diagram of the compounds of the present invention, * compared with the solvent group p < 0.05, # compared with the normal group p < 0.05;

[0110] Fig.14 This is the effect diagram of the compounds of the present invention in treating dry eye, ** compared with the solvent group p<0.01, ## compared with the normal group p<0.01, $ compared with the cyclosporine eye drops group p<0.05;

[0111] Fig.15 This is a graph showing 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, * compared with the solvent group p<0.05, # compared with the normal group p<0.05, $ compared with the cyclosporine eye drops group p<0.05;

[0112] Fig.16This is a graph showing the effect of the compounds of the present invention on reducing the level of inflammatory factors in the corneal tissue of dry eye mice, * compared with the solvent group p<0.05, # compared with the normal group p<0.05, $ compared with the cyclosporine eye drops group p<0.05;

[0113] Fig.17 This is a graph showing the effect of the compounds of the present invention on reducing retinal vascular permeability in diabetic mice, * compared with the solvent group p<0.05, # compared with the normal group p<0.05, $ compared with the t-AUCB group p<0.05;

[0114] Fig.18 This is a diagram showing the effect of the compounds of the present invention on protecting the retinal vascular homeostasis of diabetic mice, * compared with the solvent group p < 0.05, # compared with the normal group p < 0.05, $ compared with the t-AUCB group p < 0.05; ↑ indicates free pericytes, ▲ indicates acellular capillaries;

[0115] Fig.19 The distribution results of the compounds of the present invention in the eye tissues of Dutch rabbits are shown in FIG.

[0116] Fig. 20 This is a graph showing the effects of the compounds of the present invention on the contents of 14,15-EET and 14,15-DHET in rats. *p<0.05 compared with the EC5026 group;

[0117] Fig.21 This is the molecular docking result diagram of EC5026 and sEH;

[0118] Fig. 22 This is a diagram showing the molecular docking results of the compound of Example 1 of the present invention and sEH;

[0119] Fig.23 This is a diagram showing the molecular docking results of the compound of Example 3 of the present invention and sEH;

[0120] Fig.24 This is a diagram showing the molecular docking results of the compound of Example 16 of the present invention and sEH. DETAILED DESCRIPTION

[0121] In the following examples, methods for preparing some of the compounds are described. It should be understood that the following methods and other methods known to those of ordinary skill in the art can be applied to the preparation of all compounds described in the present invention. The examples are intended to illustrate rather than limit the scope of the present invention.

[0122] The present invention relates to a series of 2-substituted aminobenzothiazole compounds, which have been 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 therapeutic effects on inflammation, stroke, diabetes, fibrosis, pain, depression and dry eye.

[0123] The H NMR spectra of the compounds were measured using Bruker ARX-400 or Bruker ARX-600, and the mass spectra were measured using Agilent 1100 LC / MSD; all reagents used were of analytical or chemical purity.

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

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

[0126] At room temperature, 25.5 g (130.6 mmol) of 2-amino-5-nitrobenz[d]thiazole (intermediate A, commercially available) and 16.0 g (130.6 mmol) of 4-dimethylaminopyridine (DMAP) were added to 255 mL of dichloromethane in sequence, and the temperature was raised to 40°C. 56.9 g (261.2 mmol) of (Boc)2O was slowly added dropwise to the reaction solution, and the reaction was continued for 2 h. After the reaction, the reaction solution was cooled to room temperature, evaporated to dryness under reduced pressure, and 255 mL of 10% citric acid aqueous solution was poured into the residue, stirred for 1 h, filtered, and the filter cake was washed with water and dried to obtain 31.0 g of a light yellow solid with a yield of 80.4%.

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

[0128] 1.2 Preparation of N-[2-(4-morpholinyl)ethyl]-N-tert-butyloxycarbonyl-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 to 50 mL of dioxane in sequence. After the addition, the reaction solution was heated to 90 ° C and reacted for 8 hours. After the reaction was completed, the filtrate was filtered while hot and evaporated to dryness under reduced pressure to obtain a brown oily liquid. 200 mL of ethyl acetate was added to dissolve the residual liquid, washed with water, and evaporated to dryness under reduced pressure to obtain 4.2 g of a light 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-butyloxycarbonyl-2,5-diaminobenzo[d]thiazole (D)

[0132] At room temperature, add 2.3g (41.1mmol) of reduced iron powder and 4.4g (82.4mmol) of ammonium chloride to 45mL of 95% ethanol and reflux for 10min. Add 4.2g (10.3mmol) of intermediate C to the ethanol solution in batches, and reflux for 2h. After the reaction is completed, filter while hot, and evaporate the filtrate to dryness under reduced pressure to obtain 2.5g of brown solid, with a yield of 65.2%.

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

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

[0135] Under ice-water bath, add 1.4g (11.2mmol) 4-chloroaniline and 0.7g (6.7mmol) sodium carbonate to tetrahydrofuran solution, stir, add 1.9g (11.9mmol) tetrahydrofuran (3mL) solution of phenyl chloroformate dropwise to the reaction solution, react at room temperature for 2h. After the reaction, evaporate the reaction solution under reduced pressure, add 10mL water to the residue, stir, precipitate solid, filter by suction, wash the filter cake with water, and dry to obtain 2.5g solid, yield 91%.

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

[0137] At room temperature, add 2.0g (5.3mmol) of intermediate D and 0.8mL (5.8mmol) of triethylamine to 20mL of dry dioxane and stir to dissolve. Add 1.6g (6.3mmol) of intermediate E to the reaction solution in batches. After the addition, heat the reaction solution to 55°C and react for 5h. After the reaction, cool the reaction solution to room temperature, add 20mL of water, stir, filter and dry to obtain 2.2g of 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] At room temperature, 0.19 g (0.36 mmol) of intermediate F was added to 2 mL of saturated hydrogen chloride in dioxane solution, and the reaction solution was heated to 60°C for 3 h. After the reaction, the reaction solution was cooled to room temperature and filtered to obtain the hydrochloride of the compound of Example 1. The hydrochloride of the compound of Example 1 was added to water, and the pH value was adjusted to 10-11 with 20% sodium hydroxide aqueous solution. Solid precipitated, filtered and dried to obtain 0.12 g of white solid, which was the compound of Example 1, with a yield of 79.0%.

[0140] 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] + .

[0141] With suitable R 4 Substituted 2-amino-5-(6-)nitrobenzo[d]thiazole (A) was used as a raw material to synthesize the key intermediate D according to the synthesis method of 1.1 to 1.3 in Example 1; R 3 Substituted aniline was used as the raw material to synthesize intermediate E according to the synthesis method of 1.4 in Example 1; intermediate D and intermediate E were used to prepare intermediate F according to the synthesis method of 1.5 in Example 1, and then the compounds of Examples 2 to 16 were prepared according to the synthesis method of 1.6, whose structural formulas are shown in Table 1.

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

[0143] Using 2-amino-5-nitrobenz[d]thiazole as a raw material, the key intermediate B was synthesized according to the synthesis method of 1.1 in Example 1; intermediate B and N-(2-chloroethyl)pyrrolidin-2-one were used to synthesize intermediate D according to the synthesis methods of 1.2 to 1.3 in Example 1; using 4-chloroaniline as a raw material, intermediate E was synthesized according to the synthesis method of 1.4 in Example 1; intermediate D and intermediate E were used according to the synthesis methods of 1.5 to 1.6 in Example 1 to prepare the compound of Example 2.

[0144] 1H 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] - .

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

[0146] Using 2-amino-6-nitrobenz[d]thiazole as a raw material, the key intermediate D was synthesized according to the synthesis method of 1.1 to 1.3 in Example 1; using 4-chloroaniline as a raw material, the intermediate E was synthesized according to the synthesis method of 1.4 in Example 1; intermediate D and intermediate E were synthesized according to the synthesis method of 1.5 to 1.6 in Example 1 to obtain the compound of Example 3.

[0147] 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] - .

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

[0149] Using 2-amino-6-nitrobenz[d]thiazole as a raw material, the key intermediate B was synthesized according to the synthesis method of 1.1 in Example 1; intermediate B and N,N-dimethyl-2-chloroethylamine were used to synthesize intermediate D according to the synthesis methods of 1.2 to 1.3 in Example 1; using 4-chloroaniline as a raw material, intermediate E was synthesized according to the synthesis method of 1.4 in Example 1; intermediate D and intermediate E were used according to the synthesis methods of 1.5 to 1.6 in Example 1 to prepare the compound of Example 4.

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

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

[0152] Using 2-amino-5-nitrobenzo[d]thiazole as a raw material, the key intermediate B was synthesized according to the synthesis method of 1.1 in Example 1; intermediate B and N-(2-chloroethyl)piperidine were used to synthesize intermediate D according to the synthesis methods of 1.2 to 1.3 in Example 1; using 3-fluoro-4-chloroaniline as a raw material, intermediate E was synthesized according to the synthesis method of 1.4 in Example 1; intermediate D and intermediate E were used according to the synthesis methods of 1.5 to 1.6 in Example 1 to prepare the compound of Example 5.

[0153] 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] + .

[0154] Example 6 Preparation of 1-{N-[2-(1-piperidinyl)ethyl]-2-aminobenzo[d]thiazol-5-yl}-3-(4-chlorophenyl)urea

[0155] Using 2-amino-5-nitrobenz[d]thiazole as a raw material, the key intermediate D was synthesized according to the synthesis method of 1.1 to 1.3 in Example 1; using 3-fluoro-4-chloroaniline as a raw material, the intermediate E was synthesized according to the synthesis method of 1.4 in Example 1; intermediate D and intermediate E were synthesized according to the synthesis method of 1.5 to 1.6 in Example 1 to obtain the compound of Example 6.

[0156] 1H 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] + .

[0157] Example 7 Preparation of 1-{N-[2-(4-morpholinyl)ethyl]-2-aminobenzo[d]thiazol-6-yl}-3-(4-trifluoromethoxyphenyl)urea

[0158] Using 2-amino-6-nitrobenz[d]thiazole as the raw material, the key intermediate D was synthesized according to the synthesis method of 1.1 to 1.3 in Example 1; using 4-(trifluoromethoxy)aniline as the raw material, the intermediate E was synthesized according to the synthesis method of 1.4 in Example 1; intermediate D and intermediate E were synthesized according to the synthesis method of 1.5 to 1.6 in Example 1 to obtain the compound of Example 7.

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

[0160] Example 8 Preparation of 1-{N-[2-(2-oxo-1-pyrrolidinyl)ethyl]-2-aminobenzo[d]thiazol-5-yl}-3-(3-fluoro-4-chlorophenyl)urea

[0161] Using 2-amino-5-nitrobenzo[d]thiazole as a raw material, the key intermediate B was synthesized according to the synthesis method of 1.1 in Example 1; intermediate B and N-(2-chloroethyl)pyrrolidin-2-one were used to synthesize intermediate D according to the synthesis methods of 1.2 to 1.3 in Example 1; using 3-fluoro-4-chloroaniline as a raw material, intermediate E was synthesized according to the synthesis method of 1.4 in Example 1; intermediate D and intermediate E were used according to the synthesis methods of 1.5 to 1.6 in Example 1 to prepare the compound of Example 8.

[0162] 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] + .

[0163] Example 9 Preparation of 1-{N-[2-(4-morpholinyl)ethyl]-2-aminobenzo[d]thiazol-6-yl}-3-(1-adamantyl)urea

[0164] Using 2-amino-6-nitrobenzo[d]thiazole as a raw material, the key intermediate D was synthesized according to the synthesis method of 1.1 to 1.3 in Example 1; using adamantane amine as a raw material, the intermediate E was synthesized according to the synthesis method of 1.4 in Example 1; intermediate D and intermediate E were synthesized according to the synthesis method of 1.5 to 1.6 in Example 1 to obtain the compound of Example 9.

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

[0166] Example 10 Preparation of 1-{N-[2-(2-oxo-1-pyrrolidinyl)ethyl]-2-aminobenzo[d]thiazol-6-yl}-3-(4-chlorophenyl)urea

[0167] Using 2-amino-6-nitrobenzo[d]thiazole as the raw material, the key intermediate B was synthesized according to the synthesis method of 1.1 in Example 1; intermediate B and N-(2-chloroethyl)pyrrolidin-2-one were used to synthesize intermediate D according to the synthesis methods of 1.2 to 1.3 in Example 1; using 4-chloroaniline as the raw material, intermediate E was synthesized according to the synthesis method of 1.4 in Example 1; intermediate D and intermediate E were used according to the synthesis methods of 1.5 to 1.6 in Example 1 to prepare the compound of Example 10.

[0168] 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] + .

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

[0171] 1 H NMR (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=8.8 Hz,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] + .

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

[0174] 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] + .

[0175] Example 13 Preparation of 1-{N-[2-(2-oxo-1-piperidinyl)ethyl]-2-amino-5-fluorobenzo[d]thiazol-6-yl}-3-(4-chlorophenyl)urea

[0176] Using 2-amino-5-fluoro-6-nitrobenz[d]thiazole as the raw material, the key intermediate B was synthesized according to the synthesis method of 1.1 in Example 1; intermediate B and N-(2-chloroethyl)piperidin-2-one were used to synthesize intermediate D according to the synthesis methods of 1.2 to 1.3 in Example 1; using 4-chloroaniline as the raw material, intermediate E was synthesized according to the synthesis method of 1.4 in Example 1; intermediate D and intermediate E were used according to the synthesis methods of 1.5 to 1.6 in Example 1 to prepare the compound of Example 13.

[0177] 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] + .

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

[0179] Using 2-amino-6-nitrobenzo[d]thiazole as a raw material, the key intermediate B was synthesized according to the synthesis method of 1.1 in Example 1; intermediate B and N-(2-chloroethyl)morpholine-3-one were used to synthesize intermediate D according to the synthesis methods of 1.2 to 1.3 in Example 1; using adamantane amine as a raw material, intermediate E was synthesized according to the synthesis method of 1.4 in Example 1; intermediate D and intermediate E were prepared according to the synthesis methods of 1.5 to 1.6 in Example 1.

[0180] Example 14 Compound.

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

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

[0183] Using 2-amino-5-chloro-6-nitrobenz[d]thiazole as the raw material, the key intermediate B was synthesized according to the synthesis method of 1.1 in Example 1; intermediate B and N-(2-chloroethyl)morpholine-3-one were used to synthesize intermediate D according to the synthesis methods of 1.2 to 1.3 in Example 1; using 4-chloroaniline as the raw material, intermediate E was synthesized according to the synthesis method of 1.4 in Example 1; intermediate D and intermediate E were used according to the synthesis methods of 1.5 to 1.6 in Example 1 to prepare the compound of Example 15.

[0184] 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] + .

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

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

[0187] 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-MSm / z:447.9[M+H] + .

[0188] Table 1 Structural formula of compounds of Examples 1 to 16

[0189]

[0190]

[0191] Biological activity test:

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

[0193] The 2-substituted aminobenzothiazole compounds of the general formula I of the present invention were tested for their in vitro sEH inhibitory activity using fluorescence analysis. t-AUCB was used as a control.

[0194] The sEH solution and the fluorescent substrate PHOME (2-(3-phenyloxirane-2-yl)acetic acid 1-cyano-1-(6-methoxy-2-naphthyl)methyl ester) solution were both prepared with 25mM Bis-Tris (pH 7.0) buffer. The example compounds and t-AUCB were dissolved in DMSO and diluted into a series of different concentrations. 100μL enzyme solution and 5μL test compound solution were added to the wells of a 96-well plate, incubated at 37°C for 5 minutes, and then 95μL substrate solution was added, and the incubation continued at 37°C for 15 minutes. Only the corresponding volume of Bis-Tris (pH 7.0) buffer and DMSO were added to the blank wells, and only sEH, substrate and DMSO were added to the full-living wells. After the incubation, the fluorescence intensity was measured under the conditions of excitation wavelength of 330nm and emission wavelength of 465nm. The inhibition rate of compounds of different concentrations was calculated based on the full-living wells, and the IC was calculated using SPSS software. 50 value.

[0195] IC of some example compounds and t-AUCB for sEH 50 See Table 2 for data.

[0196] Table 2 Inhibitory activity of some of the compounds in the examples on sEH

[0197] Compound <![CDATA[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 Embodiment 11 0.3 Example 12 0.3 Example 13 1.7 Embodiment 15 0.3 Example 16 0.2 t-AUCB 8.0

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

[0199] 2. Determination of the anti-inflammatory effects of the compounds of Examples 1, 3, 10, 15 and 16

[0200] 42 male Kunming mice (20-25 g) were randomly divided into 7 groups, namely, Example 1 group, Example 3 group, Example 10 group, Example 15 group, Example 16 group, celecoxib group and blank solvent group, with 6 mice in each group. 50 μL of 1% λ-carrageenan physiological saline solution was injected subcutaneously in the sole to prepare the inflammatory swelling model. 1 h before modeling, the mice were intraperitoneally injected with 100 mg / kg of the corresponding compound solution (20 mg / mL, 10% PEG 400 and 10% Tween 80 aqueous solution) or blank solvent. The initial paw thickness of the mice before modeling was taken as 100%, and the changes in the paw thickness of the mice were measured. The degree of paw swelling of the mice was statistically analyzed as follows: Figure 1 shown.

[0201] The results showed that the compounds of Example 1, Example 3, Example 10, Example 15 and Example 16 were all able to significantly inhibit carrageenan-induced mouse foot swelling, and the effects were better than celecoxib.

[0202] 3. Determination of the Anti-Stroke Effects of the Compounds of Examples 1, 3, 10, 15 and 16 3.1 Therapeutic Effects on Acute Ischemic Stroke

[0203] The middle cerebral artery occlusion (MCAO) reperfusion model was prepared in male SD rats by thread embolism. After 2 hours of ischemia, the thread embolism was removed and perfusion was restored. Immediately after the start of reperfusion, 1 mg / kg or 3 mg / kg of the above-mentioned example compound solution (1.5 mg / mL, 10% PEG 400 and 10% Tween 80 aqueous solution), t-AUCB solution (1.5 mg / mL, 10% PEG 400 and 10% Tween 80 aqueous solution), 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 hours, the rats were killed by anesthesia with 10% chloral hydrate, and the brain tissue was cut into 4-5 consecutive coronal slices at equal distances. The brain slices were completely immersed in 2% TTC (2,3,5-triphenyltetrazolium chloride) solution for staining and then photographed to calculate the infarct volume. Figure 2 shown.

[0204] The results showed that the compounds of Example 1, Example 3, Example 10, Example 15 and Example 16 could significantly reduce the cerebral infarction volume of MCAO rats, and the effect was better than 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 through the tail vein immediately after reperfusion. After 24 hours, the ischemic side brain tissue was weighed, and PBS buffer solution was added at a weight-to-volume ratio of 1:9. The tissue was cryogenically ground for 1 min using a tissue grinder, 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 kits (refer to the instructions of the ELISA kit) were used to determine the concentrations of TNF-α and IL-1β in brain tissue. Figure 3 shown.

[0206] The results showed that the compounds of Example 3 and Example 16 could significantly reduce the levels of inflammatory factors in the brain tissue of MCAO rats, and the effects were better than t-AUCB.

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

[0208] 78 male SD rats were used to prepare the MCAO reperfusion model and 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. There were 8 rats in the sham operation group and 14 rats in each of the other groups. The rats were injected into the tail vein once a day after ischemia reperfusion (experimental drugs were the same as those in 3.1) for two consecutive weeks. The survival of the animals was recorded every day. The results are shown in Table 1. Figure 4 The Morris water maze experiment was carried out in the third week. The rats were placed in the water facing the pool wall and their latency to find the platform within 60 seconds was recorded. If the rats found the platform within 60 seconds, they were guided to the platform and stayed there for 30 seconds. The latency of the rats to find the platform was recorded for 4 days to evaluate the spatial memory ability of the rats. The experimental results are shown in Figure 5 shown.

[0209] The results showed that the compounds of Example 3 and Example 16 were able to increase the survival rate of rats, significantly reduce the escape latency of rats, and enhance learning and memory abilities, and their effects were better than those of t-AUCB and edaravone.

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

[0211] Forty-eight male BALB / c mice (18-22 g) were randomly divided into eight groups, namely, a normal control group, a blank solvent group, Example 1 group, Example 3 group, Example 10 group, Example 15 group, Example 16 group and a t-AUCB group, with six 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 with a pH of 4.5) at a dose of 50 mg / kg once a day for 5 consecutive days to prepare a diabetic model. Starting from 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 follows: Figure 6 shown.

[0212] The results showed that the blood glucose of mice in the drug administration group was significantly lower than that in the blank solvent group. The compounds of Examples 1, 3, 10, 15 and 16 could alleviate STZ-induced hyperglycemia, and the compounds of Examples 3 and 16 were better than t-AUCB.

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

[0214] Forty-eight male C57BL6 / J mice (18-22 g) were randomly divided into 8 groups, namely, 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, with 6 mice in each group. After anesthetizing the mice, the left ureter was ligated and cut to prepare a unilateral ureteral obstruction (UUO) model. In the sham operation group, only the ureter was separated without ligation. Starting 3 days before modeling, the mice were intraperitoneally injected with 10 mg / kg of the corresponding compound solution (the experimental drug preparation was the same as 4) or blank solvent once a day. The animals were killed 7 days after modeling, and the kidneys were taken and Masson staining was used to detect histopathological changes. The experimental results are shown as follows. Figure 7 shown.

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

[0216] 6. Determination of analgesic effect of the compounds of Example 1, Example 3, Example 10, Example 15 and Example 16 6.1 Determination of analgesic effect

[0217] (1) Male SD rats (200-250 g) were anesthetized after the initial mechanical withdrawal threshold (MWT) of the plantar reflex was measured using a Von Frey electronic analgesic. The rats were then anesthetized and the skin was longitudinally incised on the left femur. The muscles were bluntly separated to expose the sciatic nerve trunk and its distal branches. The surrounding adhesion tissue and the three branches of the sciatic nerve: the tibial nerve, the common peroneal nerve, and the sural nerve were separated. The tibial nerve and the common peroneal nerve were tightly ligated with silk thread. At the same time, about 2-4 mm of the nerve was cut off at the distal end of the ligature to preserve and avoid injuring the sural nerve. The selective sciatic nerve branch injury (SNI) pain model was prepared. One week after the 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, namely, 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, 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 at a dose of 3 mg / kg. Taking the MWT before administration as 100%, the changes in MWT at different time points after administration were measured, and the MWT%-time curve was drawn to calculate the area under the curve (AUC). The experimental results are shown in Figure 2. Figure 8 shown.

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

[0220] The DNP model rats were divided into 7 groups, namely, 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, 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 dosage was 3 mg / kg. The MWT before administration was 100%, and the changes of MWT at different time points after administration were measured, and the MWT%-time curve was drawn, and the area under the curve (AUC) was calculated. The experimental results are shown in Figure 2. Fig. 9 shown.

[0221] (3) C57BL6 / J mice (18-22 g) were intraperitoneally injected with paclitaxel solution (0.2 mg / mL, solvent DMSO-PEG400-Tween 80-normal saline (1:1:1:7, V / V)) at 1, 3, 5, and 7 days, respectively, with an injection dose of 2 mg / kg. After the model was established, the MWT of the mice was monitored every 4 days using a Von Frey fiber analgesia instrument 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, namely, 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, and the corresponding compound solution (3.75 mg / mL, 10% PEG 400 and 10% Tween 80 aqueous solution) or blank solvent was administered by gavage, and the dosage was 30 mg / kg. Taking the MWT before administration as 100%, the changes of MWT at different time points after administration were measured, and the MWT%-time curve was drawn to calculate the area under the curve (AUC). The experimental results are shown in Figure 2. Fig.10 shown.

[0223] The results showed that the compounds of Examples 1, 3, 10, 15 and 16 could significantly improve the MWT of SNI rats, DNP rats and PINP mice and relieve neuropathic pain, and the compounds of Examples 3 and 16 were better than EC5026 and pregabalin.

[0224] 6.2 Determination of inflammatory factor levels in the spinal cord

[0225] A batch of 30 SD rats (200-250g) were randomly divided into 5 groups, namely, Example 3 group, Example 16 group, EC5026 group, blank solvent group and sham operation group, with 6 rats in each group. The SNI model was prepared according to the method under 6.1. Only the tibial nerve, common peroneal nerve and sural nerve were isolated in the sham operation group. After the model was successfully established, the corresponding compound solution (1.2mg / mL, 10% PEG400 and 10% Tween 80 aqueous solution) or blank solvent was administered by gavage, and the dosage was 3mg / kg. After 2h of administration, the animals were anesthetized and cardiac perfusion was performed with about 200mL of physiological saline and about 100mL of 4% paraformaldehyde in sequence. The L4-L6 spinal cord was taken, weighed, and PBS buffer solution was added at a weight-to-volume ratio of 1g:9mL, and homogenized at 4°C. The tissue homogenate was centrifuged at 4°C and 2500 rpm for 10min, and the supernatant was taken for use. TNF-α and IL-1β enzyme-linked immunosorbent assay kits (refer to the kit instructions) were used to measure the concentrations of inflammatory factors TNF-α and IL-1β in the rat spinal cord. Fig.11 shown.

[0226] The results showed that the compounds of Example 3 and Example 16 could significantly reduce the levels of inflammatory factors in the spinal cord of SNI rats, and the effects were better than pregabalin.

[0227] 6.3 Effects on motor coordination in rats

[0228] The effect of the compound on the motor coordination function of rats was evaluated using a rotarod 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, with 6 rats in each group. Three days before the test, the rats were trained three times a day at a speed of 10-20 r / min, each time for 5 minutes, and the interval between two training sessions was 20 minutes as fatigue recovery time. On the day of the test, the corresponding compound solution (8 mg / mL, 10% PEG 400 and 10% Tween 80 aqueous solution) or blank solvent was administered by gavage at a dose of 20 mg / kg. The walking time of the rats on the rotarod with a speed of 25 r / min was recorded before and after the administration. The walking time of the rats on the rotarod was counted as follows: Fig.12 shown.

[0229] The results showed that the walking time of rats in the pregabalin group on the rotarod was significantly reduced, 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 rotarod, and there was no adverse reaction of the central nervous system caused by pregabalin.

[0230] 7. Determination of the antidepressant effect of the compounds of Examples 1, 3, 10, 15 and 16

[0231] 42 male Kunming mice (20-25g) were randomly divided into 7 groups, namely, Example 1 group, Example 3 group, Example 10 group, Example 15 group, Example 16 group, blank solvent group and normal control group, with 6 mice in each group. The Kunming mice were pre-swimmed for 10 minutes. Except for the normal control group, the mice were intraperitoneally injected with 0.5 mg / kg lipopolysaccharide in saline solution (0.1 mg / mL). 23 hours later, 10 mg / kg of the corresponding compound solution (6 mg / mL, 10% PEG 400 and 10% Tween 80 aqueous solution) or blank solvent was subcutaneously injected. One hour later, a forced swimming test was performed for 6 minutes, and the immobility time (s) of each group of mice within the next 4 minutes was recorded. The immobility time of the mice in the forced swimming test was calculated as follows: Fig.13 shown.

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

[0233] 8. Determination of the effect of the compounds of Examples 1, 3, 10, 15 and 16 on relieving dry eye syndrome

[0234] 72 male BALB / c mice (18-22g) were randomly divided into 8 groups, namely, 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, with 9 mice in each group. Except for the normal control group, the mice were dripped with 0.2% benzalkonium chloride (BAC) physiological saline solution 3 times a day for 15 consecutive days to establish a dry eye model. The compound prepared in the above example (2 mg / mL, 10% hydroxypropyl-β-cyclodextrin solution), cyclosporine eye drops (II) (commercially available, 0.4mL: 0.2mg) or blank solvent were used for intervention treatment, 3 times a day, 5μL per eye each time, for 15 consecutive days. The tear secretion of mice and the corneal sodium fluorescein staining score were measured at different times, as shown in Figure 2. Fig.14 shown.

[0235] The results showed that the compounds of Example 1, Example 3, Example 10, Example 15 and Example 16 could significantly alleviate BAC-induced dry eye, increase tear secretion in mice, and alleviate corneal damage, and the effects were better than 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, with 15 mice in each group. The dry eye model was prepared and the drug was administered according to the above method. After 15 days, the mice were killed and the corneas were taken. Six corneas from every three mice were combined into one sample, and three samples were taken from each group for the determination of 14,15-EET levels and 14,15-DHET levels in tissues. The remaining corneas were combined into one sample for every four mice, and three samples were taken from each group for the determination of TNF-α concentration in tissues. The LC-MS / MS method was used to determine the 14,15-EET level and 14,15-DHET level, and the enzyme-linked immunosorbent assay kit was used to determine the TNF-α concentration, referring to the instructions of the kit. The 14,15-EET level and 14,15-DHET level in tissues are as follows. Fig.15 As shown in Figure 2, the concentration of inflammatory factor TNF-α in tissues is Fig.16 shown.

[0237] The results showed that the compounds of Example 3 and Example 16 were able to 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 cyclosporine eye drops (II).

[0238] 9. Determination of the therapeutic effect of the compounds of Example 3 and Example 16 on diabetic retinopathy

[0239] 80 male C57BL / 6J mice (18-22g) were randomly divided into 5 groups, namely, 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 (0.1mol / L citric acid buffer with a pH value of 4.5), with a dose of 50mg / kg, once a day, for 5 consecutive days. After one week, the random blood glucose was ≥16.7mmol / L, and the diabetes model was successfully established, and the eye drops of the compounds of Example 3 and Example 16 (1mg / mL, 10% hydroxypropyl-β-cyclodextrin solution) or blank solvent were used for intervention treatment, twice a day, and 5μL was administered to each eye each time. Diabetic mice were fed for 6 months to induce retinopathy. After 6 months, 0.1 mL of 2% Evans blue saline solution was injected into the tail vein of the mice. The solution circulated for 2 hours. After cardiac perfusion with 50 mL of saline, retinal tissue was obtained. The homogenate was centrifuged and the supernatant was obtained. The absorbance was measured at 620 nm to investigate the permeability of retinal blood vessels. Another retinal tissue was obtained by passing through 5% pepsin solution and pepsin, and the excess tissue was blown off to obtain retinal blood vessels. After PAS staining, the morphology of retinal blood vessels and pericytes was observed and the number of free pericytes in the retina was counted. The experimental results are shown in the figure. Fig.17 , Fig.18 shown.

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

[0241] The distribution of the compounds of Example 3 and Example 16 in ocular tissues was studied using Dutch rabbits. Six male Dutch rabbits (1.6-2.4 kg) were divided into two groups, with three rabbits in each group. The eye drops of the compounds of Example 3 and Example 16 were administered to each eye, with 50 μL administered to each eye. The animals were killed 0.5 h after administration, and corneal, aqueous humor, conjunctiva, iris, vitreous, choroid, retina and sclera tissue samples were immediately collected, and 20% methanol was added at a weight-volume ratio of 1:10 for homogenization. The compound content in the supernatant was determined by LC-MS / MS. The experimental results are shown in the figure. Fig.19 shown.

[0242] The results showed that the compounds of Example 3 and Example 16 were able to reach the retinal tissue at the back of the eye after eye administration.

[0243] 10. Experimental study on the effects of the compounds of Example 3 and Example 16 on the contents of 14,15-EET and 14,15-DHET in rats

[0244] Male SD rats (250-300 g) were intragastrically administered with the test compounds of Example 3 and Example 16 and EC5026 solution (8 mg / mL, 10% PEG 400 and 10% Tween 80 aqueous solution) at a dose of 20 mg / kg, with 6 rats in each group. Before administration and 15 min, 30 min, 1 h, 2 h, 4 h, and 8 h after administration, about 0.25 mL of blood was collected from the jugular sinus 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 the figure. Fig. 20 shown.

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

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

[0247] The sEH cocrystal protein (PDB code: 4OCZ) was used as the docking model. EC5026, Example 1, Example 3 and Example 16 compounds were docked with sEH using Glide 4.5.208 software.

[0248] The molecular docking results of EC5026 and sEH are shown in Fig.21 As shown, the 3-fluoro-4-(trifluoromethoxy)phenyl group in the EC5026 structure faces the hydrophobic region inside the sEH protein cavity, 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 bidentate hydrogen bonds with the Asp335 residue, and the carbonyl oxygen atom forms hydrogen bonds with the Tyr383 and Tyr466 residues respectively; the (S)-2-methylbutanamide fragment extends to the solvent region.

[0249] The molecular docking results of the compound in Example 1 and sEH are as follows Fig. 22 As shown, the 4-chlorophenyl group in its structure faces 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 with Gln384; the two NH groups of the urea fragment form bidentate hydrogen bonds with the Asp335 residue, and the oxygen atoms of the urea fragment form hydrogen bonds with the Tyr383 and Tyr466 residues respectively; the (4-morpholinyl)ethyl part faces the solvent region.

[0250] The molecular docking results of the compound in Example 3 and sEH are as follows Fig.23 As shown, the 4-chlorophenyl group in its structure faces 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 NH groups of the urea fragment form bidentate hydrogen bonds with the Asp335 residue, and the oxygen atoms of the urea fragment form hydrogen bonds with the Tyr383 and Tyr466 residues respectively; the (4-morpholinyl)ethyl part faces the solvent region.

[0251] The molecular docking results of Example 16 compound and sEH are as follows Fig.24 As shown, the 4-chlorophenyl group in its structure faces 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 with the Asp335 residue; the two NH groups of the urea fragment form bidentate hydrogen bonds with the Asp335 residue, and the oxygen atoms of the urea fragment form hydrogen bonds with the Tyr383 and Tyr466 residues, respectively; the (2-oxo-1-pyrrolidinyl)ethyl part faces the solvent region, and the keto carbonyl group forms an additional hydrogen bond with the Ile363 residue.

[0252] Compared with EC5026, the compounds of Example 1, Example 3 and Example 16 not only maintain the key interaction force equivalent to that of EC5026 with the sEH protein, but also their benzothiazole structures generate additional interaction forces with sEH, enabling them to better bind to the sEH protein, thereby exhibiting better inhibitory activity against sEH.

[0253] In summary, in the compounds of the general formula I of the present invention, the substituent R 1 , R 2 , R 3 , R 4 , m, n, X, Y, etc., so that it can better bind to sEH, resulting in a good inhibitory effect on sEH and a 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 can achieve unexpected effects.

Claims

1. A 2-substituted aminobenzothiazole compound, characterized in that: The 2-substituted aminobenzothiazole compounds are compounds as shown in the general formula I and pharmaceutically acceptable salts thereof, in, When Y represents an N atom, X represents an S atom; when Y represents an S atom, X represents an N atom; R 1 and R 2 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, they form a 4-10 membered heterocyclic group, wherein the heterocyclic group, in addition to the nitrogen atom, optionally contains 0-3 heteroatoms selected from N, O or S, and the heterocyclic group may also be optionally replaced by 1-3 identical or different R 5 Substitution, the heterocyclic group contains 0-2 carbon-carbon double bonds; R 5 is a hydrogen atom, =0, a hydroxyl group, an amino group, a halogen group, a carboxyl group, a cyano group, a nitro group, a (C1-C6) alkyl group, a (C1-C6) alkoxy group, a (C3-C7) cycloalkyl group, a (C1-C6) alkylsulfinyl group, a (C1-C6) alkylsulfonyl group or a (C1-C6) alkylformyl group; R 3 (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 be optionally replaced by 1-3 identical or different R 6 replace; R 6 is a hydrogen atom, a hydroxyl group, a halogen group, a cyano group, a mercapto group, a carboxyl group, a carbamoyl group, a substituted or unsubstituted (C1-C6) alkyl group, a substituted or unsubstituted (C1-C6) alkoxy group, a substituted or unsubstituted (C1-C6) alkylthio group, a substituted or unsubstituted (C2-C6) alkenyl group, a substituted or unsubstituted (C1-C6) alkylsulfinyl group, a substituted or unsubstituted (C1-C6) alkylsulfonyl group, a substituted or unsubstituted (C1-C6) alkylformyl group, a substituted or unsubstituted (C1-C6) alkoxyformyl group, or -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 are the same or different substituted or unsubstituted (C1-C6) alkyl groups; the substituents of the above-mentioned substitutions are selected from hydroxyl, amino or halogen; the substitutions are mono- or poly-substituted, and the poly-substituted substituents are the same or different; R 4 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: Among the compounds, When Y represents an N atom, X represents an S atom; when Y represents an S atom, X represents an N atom; R 1 and R 2 Together with the nitrogen atom to which they are attached, they form a 4-10 membered heterocyclic group, wherein the heterocyclic group, in addition to the nitrogen atom, optionally contains 0-3 heteroatoms selected from N, O or S, and the heterocyclic group may be optionally substituted by 1-3 identical or different R 5 Substitution, the heterocyclic group contains 0-2 carbon-carbon double bonds; R 5 is a hydrogen atom, =0, a hydroxyl group, an amino group, a halogen group, a carboxyl group, a cyano group, a nitro group, a (C1-C4) alkyl group, a (C1-C4) alkoxy group, a (C3-C6) cycloalkyl group, a (C1-C4) alkylsulfinyl group, a (C1-C4) alkylsulfonyl group or a (C1-C4) alkylformyl group; R 3 is phenyl, 5-6 membered heteroaryl or adamantyl, wherein the heteroaryl contains 1-3 heteroatoms selected from N, O or S, and the phenyl or heteroaryl may be optionally substituted by 1-3 identical or different R 6 replace; R 6 is a hydrogen atom, a hydroxyl group, a halogen group, a cyano group, a mercapto group, a carboxyl group, a carbamoyl group, a substituted or unsubstituted (C1-C4) alkyl group, a substituted or unsubstituted (C1-C4) alkoxy group, a substituted or unsubstituted (C1-C4) alkylthio group, a substituted or unsubstituted (C2-C4) alkenyl group, a substituted or unsubstituted (C1-C4) alkylsulfinyl group, a substituted or unsubstituted (C1-C4) alkylsulfonyl group, a substituted or unsubstituted (C1-C4) alkylformyl group, a substituted or unsubstituted (C1-C4) alkoxyformyl group, or -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 are the same or different substituted or unsubstituted (C1-C4) alkyl groups; the substituents of the above-mentioned substitutions are selected from hydroxyl, amino or halogen; the substitutions are mono- or poly-substituted, and the poly-substituted substituents are the same or different; R 4 is 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 an N atom, X represents an S atom; when Y represents an S atom, X represents an N atom; R 1 and R 2 Together with the nitrogen atoms to which they are attached, they form R 3 is phenyl or adamantyl; the phenyl group is optionally substituted by 1-3 identical or different R 6 replace; R 6 is halogen, unsubstituted or optionally substituted by 1 to 3 identical or different halogen (C1-C4) alkyl or (C1-C4) alkoxy; R 4 is 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 an N atom, X represents an S atom; When Y represents an S atom, X represents an N atom, and R 4 is H; R 1 and R 2 Together with the nitrogen atoms to which they are attached, they form R 3 is a phenyl group, wherein the para position of the phenyl group is replaced by R 6 replace; R 6 is halogen, trifluoromethyl or trifluoromethoxy; R 4 is 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 a N atom, X represents a S atom, which is represented by the general formula (II): R 1 and R 2 Together with the nitrogen atoms to which they are attached, they form R 3 is a phenyl group, wherein the para position of the phenyl group is replaced by R 6 replace; R 6 is halogen, trifluoromethyl or trifluoromethoxy; R 4 is 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]thiazol-5-yl}-3-(4-chlorophenyl)urea; 1-{N-[2-(2-oxo-1-pyrrolidinyl)ethyl]-2-aminobenzo[d]thiazol-5-yl}-3-(4-chlorophenyl)urea; 1-{N-[2-(4-morpholinyl)ethyl]-2-aminobenzo[d]thiazol-6-yl}-3-(4-chlorophenyl)urea; 1-{N-[2-(Dimethylamino)ethyl]-2-aminobenzo[d]thiazol-6-yl}-3-(4-chlorophenyl)urea; 1-{N-[2-(4-morpholinyl)ethyl]-2-aminobenzo[d]thiazol-5-yl}-3-(3-fluoro-4-chlorophenyl)urea; 1-{N-[2-(1-piperidinyl)ethyl]-2-aminobenzo[d]thiazol-5-yl}-3-(4-chlorophenyl)urea; 1-{N-[2-(4-morpholinyl)ethyl]-2-aminobenzo[d]thiazol-6-yl}-3-(4-trifluoromethoxyphenyl)urea; 1-{N-[2-(2-oxo-1-pyrrolidinyl)ethyl]-2-aminobenzo[d]thiazol-5-yl}-3-(3-fluoro-4-chlorophenyl)urea; 1-{N-[2-(4-morpholinyl)ethyl]-2-aminobenzo[d]thiazol-6-yl}-3-(1-adamantyl)urea; 1-{N-[2-(2-oxo-1-pyrrolidinyl)ethyl]-2-aminobenzo[d]thiazol-6-yl}-3-(4-chlorophenyl)urea; 1-{N-[2-(3-oxo-4-morpholinyl)ethyl]-2-aminobenzo[d]thiazol-5-yl}-3-(3,4-dichlorophenyl)urea; 1-{N-[2-(3-oxo-4-morpholinyl)ethyl]-2-amino-5-fluorobenzo[d]thiazol-6-yl}-3-(4-chlorophenyl)urea; 1-{N-[2-(2-oxo-1-piperidinyl)ethyl]-2-amino-5-fluorobenzo[d]thiazol-6-yl}-3-(4-chlorophenyl)urea; 1-{N-[2-(3-oxo-4-morpholinyl)ethyl]-2-aminobenzo[d]thiazol-6-yl}-3-(1-adamantyl)urea; 1-{N-[2-(3-oxo-4-morpholinyl)ethyl]-2-amino-5-chlorobenzo[d]thiazol-6-yl}-3-(4-chlorophenyl)urea; 1-{N-[2-(2-oxo-1-pyrrolidinyl)ethyl]-2-amino-5-fluorobenzo[d]thiazol-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 of the 2-substituted aminobenzothiazole compound and an acid, wherein the acid is 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 and pharmaceutically acceptable salts thereof according to any one of claims 1 to 7 and a pharmaceutically acceptable carrier.

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

10. The use according to claim 9, characterized in that: The 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 eyes.

11. The use according to claim 10, characterized in that: 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 diabetic complications include diabetic retinopathy, diabetic uveitis, diabetic cataracts, diabetic nephropathy, diabetic skin diseases and diabetic peripheral neuropathy; the diabetes-related diseases include hyperlipidemia, hyperuria Acidemia 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 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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