A kind of 3-urea group substitution thiophene [2, 3-b] pyridine compound and its preparation method and application

Abstract

The application discloses a novel 3-urea-substituted thieno[2,3-b]pyridine compound, a preparation method and application thereof. The preparation method is that 5-bromothieno[2,3-b]pyridine-3-amine is subjected to nucleophilic substitution reaction with phenyl chloroformate or phenyl thiophosphonic chloride to generate an intermediate, the intermediate is subjected to nucleophilic substitution reaction with a substituted amine to generate a key intermediate, then the key intermediate is subjected to palladium-catalyzed Suzuki coupling reaction or Buchwald-Hartwig coupling reaction under alkaline conditions with a substituted aryl boronic acid or borate or a substituted arylamine to obtain a target product, namely the 3-urea-substituted thieno[2,3-b]pyridine compound. The 3-urea-substituted thieno[2,3-b]pyridine compound has a certain prospect as a DRAK2 inhibitor in the development of metabolic syndrome treatment drugs.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of pharmacy, and relates to a novel 3-ureido-substituted thieno[2,3-b]pyridine compound and a preparation method and application thereof. BACKGROUND

[0002] DRAK2 (Death Associated Apoptosis Inducing Protein Kinase 2), also known as serine / threonine kinase 17B (STK17B), belongs to the death associated protein kinase (DAPK) family (J. Biol. Chem. 1998, 273, 29066-29071; Mol. Cell. Biol. 1998, 18, 1642-1651). DRAK2 is highly expressed in immune system related tissues such as thymus, spleen, lymph node, and also has a certain degree of expression in other tissues and organs such as the olfactory lobe of the brain, the ventricular region of the brain, the hippocampus, intestinal epithelial cells, and the pancreas (J. Immunol. 2009, 182, 4762-4770; Front. Pharmacol. 2022, 13, 1014508). Recent studies have found that DRAK2 protein is abnormally highly expressed in the pancreatic tissues of hyperglycemic humans and diabetic monkeys, and that the DRAK2 protein level is increased, autophagy is blocked, and apoptosis is increased in the pancreas of diabetic model mice (db / db mice). By constructing DRAK2-cKO mice with specific knockout of DRAK2 in islet beta cells, it was found that DRAK2-cKO mice could resist islet function damage induced by a high-fat diet. Another study found that DRAK2 expression was significantly increased in the liver puncture tissues of NAFLD / NASH model mice and patients, and was positively correlated with the NASH pathological score.

[0003] Molecular mechanism studies show that DRAK2 has a direct protein-protein interaction with the key protein ULK1 of the autophagy initiation complex in pancreatic beta cells, and by directly phosphorylating the ULK1-Ser56 site, it further promotes the ubiquitination and degradation of ULK1 to negatively regulate autophagy, mitochondrial mass and beta cell function (Sci. Transl. Med. 2024, 16, eade8647). In liver cells, DRAK2 has a direct protein-protein interaction with the RNA splicing factor SRSF6, thereby causing abnormal alternative splicing of mitochondrial function-related genes, including mtDNA polymerase POLy2, which is a key factor in the development of non-alcoholic fatty liver disease (Cell. Metab. 2021, 33, 2004-2020). Therefore, given the potential of DRAK2 in the treatment of metabolic syndromes such as diabetes and non-alcoholic fatty liver disease, it is of great value to develop new structural DRAK2 small molecule inhibitors, and it is particularly urgent to obtain small molecules with high inhibitory activity and safety and effectiveness for DRAK2. SUMMARY

[0004] To solve the problems existing in the prior art, the purpose of the present application is to provide a novel 3-ureido-substituted thieno[2,3-b]pyridine compound, which can be used as a small molecule inhibitor of DRAK2 and can be used for the development of drugs for the treatment of metabolic syndromes, including but not limited to: diabetes, non-alcoholic fatty liver disease, hyperlipidemia, obesity, and biliary cirrhosis.

[0005] The present application provides a 3-ureido-substituted thieno[2,3-b]pyridine compound or a stereoisomer, a pharmaceutically acceptable salt, a prodrug or a solvate thereof, the structure of the compound is as shown in the following formula (I):

[0006]

[0007] wherein,

[0008] Ring A is selected from the group consisting of phenyl, 5-9 membered heteroaryl;

[0009] R1is selected from the group consisting of -C(O)ORa, -O(O)CCH3, -S(O)2CH3, -NHS(O)2Rb, -S(O)2NH2, -C(O)CH3, -C(O)N(Ra)2, -NHC(O)Rb, substituted or unsubstituted 5-6 membered heteroaryl, substituted or unsubstituted 5-7 membered heterocyclyl; wherein each Ra is independently selected from the group consisting of hydrogen, cyclopropyl, C1-C6 alkyl; Rb is selected from the group consisting of substituted or unsubstituted C1-C6 alkyl, phenyl; R2is selected from the group consisting of hydrogen, hydroxyl, amino, halogen, C1-C6 alkyl, C1-C6 alkoxy; or, R1, R2on adjacent ring atoms of ring A together with the adjacent ring atoms form a substituted or unsubstituted 5-7 membered heterocyclyl;

[0010] R3is selected from the group consisting of cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, phenyl, pyridyl, substituted or unsubstituted benzyl,

[0011] R4is selected from the group consisting of hydrogen, methyl;

[0012] L is selected from the group consisting of a covalent single bond, -NH-;

[0013] X is selected from the group consisting of oxygen, sulfur;

[0014] Unless specifically indicated, the above-mentioned substituents of the groups mentioned involve substitution of one to three substituents selected from the group consisting of halogen, hydroxyl, amino, C1-C3 alkyl, C1-C3 alkoxy, acetyl,

[0015] In another preferred embodiment, ring A is selected from the group consisting of phenyl, pyridyl.

[0016] In another preferred embodiment, R1is selected from the group consisting of -C(O)ORa, -O(O)CCH3, -S(O)2CH3, -NHS(O)2Rb, -S(O)2NH2, -C(O)CH3, -C(O)N(Ra)2, -NHC(O)Rb, wherein each Ra is independently selected from the group consisting of hydrogen, methyl, ethyl, propyl, cyclopropyl, isopropyl, tert-butyl; Rb is selected from the group consisting of methyl, chloromethyl, ethyl, phenyl.

[0017] In another preferred embodiment, R2is selected from the group consisting of hydrogen, hydroxyl, fluorine, chlorine, methyl, methoxy.

[0018] In another preferred embodiment, R3is selected from the group consisting of cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, phenyl, pyridyl, benzyl,

[0019] In another preferred embodiment, R4 is selected from the group consisting of hydrogen.

[0020] In another preferred embodiment, L is selected from the group consisting of a covalent single bond.

[0021] In another preferred embodiment, X is selected from the group consisting of oxygen.

[0022] In another preferred embodiment, the compound of formula (I) has the structure as shown in formula (II-1) and (II-2) below:

[0023]

[0024] wherein, the definitions of ring A, R1, R2, R3, R4, and X are the same as those in formula (I).

[0025] In another preferred embodiment, the compound of formula (I) has the structure as shown in formula (III-1) and (III-2) below:

[0026]

[0027] wherein, the definitions of R1, R2, and R3 are the same as those in formula (I).

[0028] In another preferred embodiment, the compound of formula (I) is selected from the group consisting of:

[0029]

[0030] The present application also provides a preparation method of a 3-ureido-substituted thieno[2,3-b]pyridine compound (as shown in formula (I)), which comprises the following route:

[0031] (1) in the presence of a base, 5-bromothieno[2,3-b]pyridine-3-amine and phenyl chloroformate or phenyl thiocyanate undergo a nucleophilic substitution reaction in a first solvent to generate an intermediate IV-1;

[0032] (2) in the presence of a base, the intermediate IV-1 obtained in step (1) and a substituted amine undergo a nucleophilic substitution reaction in a second solvent to generate an intermediate IV-2;

[0033] (3) in the presence of a base and a palladium catalyst, the intermediate IV-2 obtained in step (2) and a substituted aryl boronic acid or boronic ester undergo a palladium-catalyzed Suzuki coupling reaction under basic conditions in a third solvent to generate a target compound II-1;

[0034] or,

[0035] In the fourth solvent, the intermediate IV-2 obtained in the step (2) is subjected to a palladium-catalyzed Buchwald-Hartwig coupling reaction with a substituted aniline under basic conditions in the presence of a base, a palladium catalyst, and a phosphine ligand, to generate the target compound II-2.

[0036] The reaction process of the route is shown in the following reaction formula (a):

[0037]

[0038] wherein R5 is B(OH)2 or B(Pin); the definitions of ring A, R1, R2, R3, R4, and X are the same as those in formula (I).

[0039] In the step (1), the first solvent is one or more of dichloromethane, tetrahydrofuran, dimethyl sulfoxide, etc.; preferably, tetrahydrofuran.

[0040] In the step (1), the ratio of the first solvent to 5-bromothieno[2,3-b]pyridine-3-amine (milliliter: millimole) is 1:0.02 to 1:0.04; preferably, 1:0.03.

[0041] In the step (1), the molar ratio of 5-bromothieno[2,3-b]pyridine-3-amine to phenyl chloroformate or phenyl thiochloroformate is 1:1 to 1:1.5; preferably, 1:1.5.

[0042] In the step (1), the base is one or more of potassium carbonate, sodium bicarbonate, etc.; preferably, sodium bicarbonate.

[0043] In the step (1), the molar ratio of the base to 5-bromothieno[2,3-b]pyridine-3-amine is 1:1 to 3:1; preferably, 1.5:1.

[0044] In the step (1), the temperature of the reaction is 20°C to 40°C; preferably, 25°C.

[0045] In the step (1), the time of the reaction is 1 to 2 hours; preferably, 1 hour.

[0046] In the step (2), the second solvent is one or more of dimethyl sulfoxide, acetonitrile, toluene, etc.; preferably, dimethyl sulfoxide.

[0047] In the step (2), the ratio of the second solvent to intermediate IV-1 (milliliter: millimole) is 1:0.02 to 1:0.04; preferably, 1:0.03.

[0048] In the step (2), the substituted amine includes one or more of cyclopentylamine, cyclohexylamine, cycloheptylamine, 4-aminotetrahydropyran, cyclopropylamine, aniline, benzylamine, trans-4-aminocyclohexanol, cyclohexylmethylamine, and the like, and is preferably cyclohexylamine.

[0049] In the step (2), the molar ratio of the intermediate IV-1 to the substituted amine is 1:1 to 1:1.5, and is preferably 1:1.5.

[0050] In the step (2), the base is one or more of potassium carbonate, triethylamine, and the like, and is preferably triethylamine.

[0051] In the step (2), the molar ratio of the base to the intermediate IV-1 is 1:1 to 3:1, and is preferably 3:1.

[0052] In the step (2), the temperature of the reaction is 20°C to 40°C, and is preferably 25°C.

[0053] In the step (2), the time of the reaction is 1 to 2 hours, and is preferably 1 hour.

[0054] In the step (3), the third solvent is one or more of dimethyl sulfoxide, N,N-dimethylformamide, toluene, water, and the like, and is preferably a mixed solvent of N,N-dimethylformamide and water.

[0055] In the step (3), the ratio of the third solvent to the intermediate IV-2 (mL:mmol) is 1:0.01 to 1:0.03, and is preferably 1:0.02.

[0056] In the step (3), the substituted aryl boronic acid or boronic ester includes one or more of p-carbamoylphenyl boronic acid, p-carboxyphenyl boronic acid, 1-isoindolin-5-boronic acid pinacol ester, 5-carboxy-2-fluorophenyl boronic acid, p-acetylamidophenyl boronic acid, 4-(N-methylformamido)phenyl boronic acid, 4-(aminosulfonyl)phenyl boronic acid, 2-methoxy-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide, 3-chloro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide, 5-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-1H-tetrazole, 4-methylsulfonamidophenyl boronic acid, 2-acetylamino pyridine-5-boronic acid, p-propionamidophenyl boronic acid, (1-acetylaminoindolin-5-yl)boronic acid, 3-ethoxycarbonylphenyl boronic acid, and the like, and is preferably p-carbamoylphenyl boronic acid.

[0057] In the step (3), the molar ratio of the intermediate IV-2 to the substituted aryl boronic acid or boronic ester is 1:1 to 1:5; preferably, 1:1.

[0058] In the step (3), the palladium catalyst is one or more of tetrakis(triphenylphosphine)palladium, dichlorobis(triphenylphosphine)palladium(II), 1,1'-bis(diphenylphosphino) ferrocene palladium(II) dichloromethane complex, and the like; preferably, tetrakis(triphenylphosphine)palladium.

[0059] In the step (3), the molar ratio of the intermediate IV-2 to the palladium catalyst is 1:0.05 to 1:0.10; preferably, 1:0.10.

[0060] In the step (3), the base is one or more of potassium carbonate, sodium carbonate, and the like; preferably, potassium carbonate.

[0061] In the step (3), the molar ratio of the base to the intermediate IV-2 is 2:1 to 3:1; preferably, 2:1.

[0062] In the step (3), the reaction temperature of the Suzuki coupling reaction is 80°C to 100°C; preferably, 85°C.

[0063] In the step (3), the reaction time of the Suzuki coupling reaction is 5 to 12 hours; preferably, 12 hours.

[0064] or,

[0065] In the step (3), the fourth solvent is one or more of toluene, dioxane, t-butanol, and the like; preferably, t-butanol. In the step (3), the ratio of the fourth solvent to the intermediate IV-2 (mL:mmol) is 1:0.03 to 1:0.05; preferably, 1:0.04.

[0066] In the step (3), the substituted aryl amine includes one or more of p-aminobenzamide, methyl 4-aminobenzoate, and the like; preferably, p-aminobenzamide.

[0067] In the step (3), the molar ratio of the intermediate IV-2 to the substituted aryl amine is 1:1 to 1:5; preferably, 1:1.

[0068] In the step (3), the palladium catalyst is one or more of tris(dibenzylideneacetone)dipalladium, tetrakis(triphenylphosphine)palladium, dichlorobis(triphenylphosphine)palladium(II), and the like; preferably, tris(dibenzylideneacetone)dipalladium.

[0069] In the step (3), the molar ratio of the intermediate IV-2 to the palladium catalyst is 1:0.05 to 1:0.10; preferably, 1:0.05.

[0070] In the step (3), the phosphine ligand is one or more of (S)-(-)-1,1'-binaphthalene-2,2'-diphenylphosphine, 4,5-(bis-diphenylphosphino)-9,9-dimethylxanthene, 2-dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl, and the like; preferably, 2-dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl.

[0071] In the step (3), the molar ratio of the intermediate IV-2 to the phosphine ligand is 1:0.05 to 1:0.10; preferably, 1:0.05.

[0072] In the step (3), the base is one or more of cesium carbonate, potassium tert-butoxide, and the like; preferably, cesium carbonate.

[0073] In the step (3), the molar ratio of the base to the intermediate IV-2 is 2:1 to 3:1; preferably, 3:1.

[0074] In the step (3), the reaction temperature of the Buchwald-Hartwig coupling reaction is 80°C to 100°C; preferably, 100°C.

[0075] In the step (3), the reaction time of the Buchwald-Hartwig coupling reaction is 5 to 12 hours; preferably, 12 hours.

[0076] In one embodiment, the method for preparing the 3-ureido-substituted thieno[2,3-b]pyridine compound (as shown in formula (I)) comprises the following steps:

[0077] (1) A nucleophilic substitution reaction is carried out at 25°C for 1 to 2 hours using 5-bromothieno[2,3-b]pyridine-3-amine as a raw material, phenyl chloroformate or phenyl thiocyanate as a reagent, sodium bicarbonate as a base, and tetrahydrofuran as a solvent, to obtain the intermediate IV-1.

[0078] (2) A nucleophilic substitution reaction is carried out at 25°C for 1 to 2 hours using the intermediate IV-1 and a substituted amine as reagents, triethylamine as a base, and dimethyl sulfoxide as a solvent, to obtain the intermediate IV-2.

[0079] (3) intermediate IV-2 and substituted arylboronic acid or boronic ester in a molar ratio of 1:1, potassium carbonate as a base, N,N-dimethylformamide and water as a solvent, tetraphenylphosphine palladium (10 mol%) catalyzed Suzuki coupling reaction at 85°C for 5-12 hours to generate the target compound II-1;

[0080] or,

[0081] intermediate IV-2 and substituted arylamine in a molar ratio of 1:1, cesium carbonate as a base, t-butanol as a solvent, 2-dicyclohexylphosphine-2',4',6'-triisopropylbiphenyl as a phosphine ligand, tris(dibenzylideneacetone)dipalladium (5 mol%) catalyzed Buchwald-Hartwig coupling reaction at 100°C for 5-12 hours to generate the target compound II-2.

[0082] The process described in the above preparation method is shown in reaction formula (b):

[0083]

[0084] wherein, R5 is B(OH)2 or B(Pin); the definitions of ring A, R1, R2, R3, R4 and X are the same as those in formula (I).

[0085] The present application also provides a medicine / pharmaceutical composition comprising the compound as shown in formula (I), formula (II-1), formula (II-2), formula (III-1), formula (III-2) or its stereoisomer, pharmaceutically acceptable salt, prodrug or solvate as described above; and a pharmaceutically acceptable carrier.

[0086] Further, the medicine / pharmaceutical composition is used alone and / or in combination with other medicines.

[0087] Preferably, the pharmaceutically acceptable carrier means that they do not produce an adverse, allergic, or other untoward reaction when administered to an animal or a human as appropriate. As a pharmaceutically acceptable carrier, there can be included, but are not limited to, sugars such as lactose, glucose, and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium methyl cellulose, ethyl cellulose, and methyl cellulose; powdered tragacanth; malt; gelatin; talc; solid lubricants such as stearic acid and magnesium stearate; calcium sulfate; vegetable oils such as peanut oil, cottonseed oil, sesame oil, olive oil, corn oil, and cocoa butter; polyhydric alcohols such as propylene glycol, glycerin, sorbitol, mannitol, and polyethylene glycol; alginic acid; emulsifiers such as Tween; wetting agents such as sodium lauryl sulfate; coloring agents; flavoring agents; tabletting agents, stabilizers; antioxidants; preservatives; pyrogen-free water; isotonic salt solutions; and phosphate buffer solutions, etc. These substances are used as needed to help the stability of the formulation or to help improve the activity or its bioavailability or to produce an acceptable taste or odor in the case of oral administration.

[0088] Preferably, the pharmaceutical / drug composition can further include a physiologically compatible adjuvant including a buffer, a diluent, an excipient, a filler, a binder, a humectant, a disintegrant, an absorption enhancer, a surfactant, an adsorption carrier, a lubricant, etc.

[0089] Specifically, the pharmaceutical / drug composition is administered orally, by injection, nasally, transdermally, or mucosally, etc.

[0090] Preferably, the dosage form of the pharmaceutical / drug composition includes an injection, a sterile powder for injection, a tablet, a pill, a capsule, a lozenge, an elixir, a powder, a granule, a syrup, a solution, a tincture, an aerosol, a powder mist, or a suppository, etc. The pharmaceutical / drug composition of the above various dosage forms can be prepared according to the conventional method in the pharmaceutical field.

[0091] Specifically, the pharmaceutical / drug composition includes a liquid dosage form, a gaseous dosage form, a solid dosage form, and a semi-solid dosage form, etc.

[0092] Preferably, the route of administration of the medicament / pharmaceutical composition is preferably parenteral administration, injection administration or oral administration. The injection administration preferably includes intravenous injection, intramuscular injection, intraperitoneal injection, intradermal injection or subcutaneous injection, etc. The medicament / pharmaceutical composition is in various dosage forms conventional in the art, preferably in the form of solid, semi-solid, gas or liquid, i.e. can be aqueous solution, non-aqueous solution or suspension, more preferably tablet, capsule, granule, injection or infusion, etc. More preferably, it is administered intravascularly, subcutaneously, intraperitoneally or intramuscularly. Preferably, the medicament / pharmaceutical composition can also be administered as an aerosol or coarse spray, i.e. nasally; or intrathecally, intramedullary or intraventricularly. More preferably, the medicament / pharmaceutical composition can also be transdermally, transcutaneously, topically, enterally, intravaginally, sublingually or rectally administered. The medicament / pharmaceutical composition of the present application can be prepared into various dosage forms as needed, and the dosage beneficial to the patient can be determined by the physician according to the patient's species, age, weight and general disease condition, administration method, etc. The administration method can be injection or other treatment, for example.

[0093] The dosage level of the medicament / pharmaceutical composition of the present application can be adjusted according to the amount of composition required to achieve the desired diagnostic or therapeutic result. The administration regimen can also be a single injection or multiple injections, or adjusted. The selected dosage level and regimen are reasonably adjusted depending on various factors including the activity and stability (i.e. half-life) of the cell medicament / pharmaceutical composition, the preparation, the administration route, the combination with other drugs or treatments, the disease or disorder to be detected and / or treated, and the health condition and previous medical history of the subject to be treated, etc.

[0094] The therapeutically effective dose of the medicament / pharmaceutical composition of the present application can be initially estimated in cell culture assays or animal models, such as rodents, monkeys, dogs, pigs and / or primates. Animal models can also be used to determine the appropriate administration concentration range and route. Subsequently, it can be used to determine the useful dose and route of administration in humans. In general, the determination and adjustment of effective amount or dose and the evaluation of when and how to make such adjustments are known to those skilled in the art.

[0095] For further guidance on formulations, dosages, administration regimens and measurable therapeutic outcomes, see Berkow et al. (2000) The Merck Manual of Medical Information and Merck & Co. Inc., Whitehouse Station, New Jersey; Ebadi (1998) CRC Desk Reference of Clinical Pharmacology, among others.

[0096] The present application also provides the use of the compound as shown in formula (I), formula (II-1), formula (II-2), formula (III-1), formula (III-2), formula (1) to formula (26), or a stereoisomer, a pharmaceutically acceptable salt, a prodrug or a solvate thereof, or a preparation method as described above, or a drug / drug composition as described above in the preparation of a drug for treating or preventing a disease related to the activity or expression amount of DRAK2, or in the preparation of a drug for inhibiting the activity of DRAK2 kinase.

[0097] In another preferred embodiment, the disease is metabolic syndrome, etc.; wherein the metabolic syndrome includes but is not limited to diabetes, non-alcoholic fatty liver disease, hyperlipidemia, obesity, biliary cirrhosis.

[0098] It should be understood that, within the scope of the present application, each of the technical features described above and in the following (such as the examples) can be combined with each other to form a new or preferred technical solution. Due to the limited space, they will not be repeated one by one here.

[0099] The beneficial effects of the present application are embodied in that the 3-ureido-substituted thieno[2,3-b]pyridine compounds prepared by the present application have novel structures, good DRAK2 inhibitory activity, and can significantly promote the secretion of insulin by pancreatic beta cells, and have certain application prospects in the discovery of drugs for metabolic syndrome treatment, and can be used for developing new drugs for metabolic syndrome treatment. DETAILED DESCRIPTION

[0100] The present application will be further described in conjunction with the following specific examples. The process, conditions, implementation methods, etc. of the present application are all general knowledge and common sense in the art, and the present application has no special limitations.

[0101] The following description of at least one exemplary embodiment is merely exemplary in nature and is in no way intended to limit the application or its application or uses. Based on the examples in the present application, all other examples obtained by those of ordinary skill in the art without creative work are within the scope of protection of the present application.

[0102] In all examples shown and discussed here, any specific value should be interpreted as merely exemplary, not as a limitation. Therefore, other examples of exemplary embodiments can have different values.

[0103] The application provides a novel 3-ureido-substituted thieno[2,3-b]pyridine compound and a preparation method and application thereof. The preparation method is that 5-bromothieno[2,3-b]pyridine-3-amine and phenyl chloroformate or phenyl thiophosphonic chloride are subjected to nucleophilic substitution reaction to generate an intermediate, the intermediate and substituted amine are subjected to nucleophilic substitution reaction to generate a key intermediate, and then the key intermediate and substituted aryl boronic acid or borate or substituted arylamine are subjected to palladium-catalyzed Suzuki coupling reaction or Buchwald-Hartwig coupling reaction under alkaline conditions to obtain the target product 3-ureido-substituted thieno[2,3-b]pyridine compound. The 3-ureido-substituted thieno[2,3-b]pyridine compound has a certain prospect as a DRAK2 inhibitor for preparing a metabolic syndrome treatment drug.

[0104] The test materials used in the examples are all conventional biochemical reagents unless otherwise specified.

[0105] Example 1: Synthesis of compound 1.

[0106]

[0107] To a solution of 5-bromothieno[2,3-b]pyridine-3-amine (70 mg, 0.31 mmol), sodium bicarbonate (39 mg, 0.47 mmol) in tetrahydrofuran (10 mL) was added phenyl chloroformate (70 mg, 0.45 mmol), and the system was stirred at room temperature for 1 hour. The reaction was monitored by TLC until completion, and the system was rotary dried to obtain a green crude intermediate IV-1 (100 mg).

[0108] The crude intermediate IV-1 (100 mg, 0.29 mmol) was added to a solution of cyclohexylamine (42 mg, 0.42 mmol), triethylamine (87.8 mg, 0.87 mmol) in dimethyl sulfoxide (10 mL), and stirred at room temperature for 1 hour. The reaction was monitored by TLC until completion, and the reaction system was added with an appropriate amount of water and ethyl acetate for extraction, and the organic phase was dried over anhydrous sodium sulfate and then was mixed with silica gel and separated and purified by column chromatography (dichloromethane:methanol, v:v = 20:1) to obtain the intermediate IV-2 (66 mg). 1 H NMR (400 MHz, DMSO-d6) δ 8.91 (s, 1H), 8.68 (s, 1H), 8.42 (s, 1H), 7.70 (s, 1H), 6.21 (s, 1H), 3.65-3.51 (m, 1H), 1.94-1.60 (m, 5H), 1.43-1.06 (m, 5H).

[0109] Into a single necked flask, N,N-dimethylformamide (20 mL), water (4 mL), IV-2 (150 mg, 0.425 mmol), potassium carbonate (117 mg, 0.850 mmol), p- carbamoylphenylboronic acid (70 mg, 0.425 mmol), tetrakis(triphenylphosphine)palladium (49 mg, 0.0425 mmol) were added successively. The system was reacted at 85 °C for 12 h. TLC was used to monitor the reaction until completion. The reaction system was added with appropriate amount of water and ethyl acetate for extraction. The organic phase was retained, dried over anhydrous sodium sulfate, and then the organic phase was mixed with silica gel and separated and purified by column chromatography (dichloromethane:methanol, v:v = 15:1) to obtain the white solid target compound 1 (34 mg). 1 H NMR (400 MHz, DMSO-d6) δ 8.98 (s, 1H), 8.96 (s, 1H), 8.52 (s, 1H), 8.10 (s, 1H), 8.07 (s, 1H), 8.05 (s, 1H), 7.92 (s, 1H), 7.90 (s, 1H), 7.70 (s, 1H), 7.47 (s, 1H), 6.25 (d, J = 7.8 Hz, 1H), 3.60 - 3.46 (m, 1H), 1.91 - 1.50 (m, 5H), 1.40 - 1.17 (m, 5H). 13 C NMR (101 MHz, DMSO-d6) δ 167.89, 158.62, 154.82, 146.10, 140.41, 134.11, 131.04, 129.95, 128.86, 127.34, 126.74, 126.56, 106.20, 48.32, 33.40, 25.69, 24.81.

[0110] Example 2: Synthesis of compound 2.

[0111]

[0112] The raw material p-carbamoylphenylboronic acid in Example 1 of the application was replaced with p-carboxyphenylboronic acid, and compound 2 was prepared by using similar experimental steps as in Example 1 of the application. 1 H NMR (400 MHz, DMSO-d6) δ 8.98 (s, 1H), 8.96 (s, 1H), 8.52 (s, 1H), 8.10 (s, 1H), 8.07 (s, 1H), 8.05 (s, 1H), 7.92 (s, 1H), 7.90 (s, 1H), 7.70 (s, 1H), 7.47 (s, 1H), 6.25 (d, J = 7.8 Hz, 1H), 3.60 - 3.46 (m, 1H), 1.91 - 1.50 (m, 5H), 1.40 - 1.17 (m, 5H). 13CNMR(101MHz,DMSO-d6)δ167.52,158.84,154.78,146.09,142.01,130.85,130.6 6,130.63,129.94,127.71,126.71,126.66,106.29,48.32,33.40,25.68,24.81.

[0113] Example 3: Synthesis of compound 3.

[0114]

[0115] In Example 1 of this invention, p-carbamoylphenylboronic acid was replaced with 5-carboxy-2-fluorophenylboronic acid, and compound 3 was prepared using experimental steps similar to those in Example 1 of this invention. 1 H NMR (400MHz, DMSO-d6) δ13.29(s,1H),8.97(s,1H),8.80(s,1H),8.41(s,1H),8.22(d,J=7.1Hz,1H),8.14–8.04( m,1H),7.73(s,1H),7.56(m,1H),6.25(d,J=7.3Hz,1H),3.57–3.48(m,1H),1.93–1.51(m,5H),1.43–1.13(m,5H). 13 C NMR(101MHz,DMSO-d6)δ166.68,163.54,161.02,158.78,154.83,147.11,132.89,132.84,132.27,132.1 7,129.88,128.57,128.51,126.56,126.10,125.96,117.41,117.18,106.42,48.31,33.36,25.67,24.79.

[0116] Example 4: Synthesis of compound 4.

[0117]

[0118] By replacing p-carbamoylphenylboronic acid in Example 1 of the present invention with p-acetaminophenylboronic acid, compound 4 was prepared using experimental steps similar to those in Example 1 of the present invention. 1H NMR (400 MHz, DMSO-d6) δ 10.12 (s, 1H), 8.87 (s, 2H), 8.36 (s, 1H), 7.82 - 7.68 (m, 4H), 7.67 (s, 1H), 6.19 (d, J = 7.2 Hz, 1H), 3.61 - 3.49 (m, 1H), 2.09 (s, 3H), 1.94 - 1.49 (m, 5H), 1.41 - 1.12 (m, 5H). 13 C NMR (101 MHz, DMSO-d6) δ 168.96, 157.71, 154.80, 145.89, 139.87, 132.19, 131.69, 129.81, 127.88, 126.72, 125.69, 119.97, 106.09, 48.33, 33.42, 25.68, 24.82, 24.55.

[0119] Example 5: Synthesis of compound 5.

[0120]

[0121] The starting material p-aminocarbonylphenylboronic acid in Example 1 of the present application was replaced by 4-(N-methylcarboxamido)phenylboronic acid, and compound 5 was prepared by using similar experimental procedures to those in Example 1 of the present application. 1 H NMR (400 MHz, DMSO-d6) δ 9.13 (s, 1H), 8.96 (s, 1H), 8.64 - 8.52 (m, 2H), 8.07 - 7.87 (m, 4H), 7.69 (s, 1H), 6.37 (d, J = 7.3 Hz, 1H), 3.60 - 3.48 (m, 1H), 2.82 (d, J = 3.6 Hz, 3H), 1.93 - 1.51 (m, 5H), 1.42 - 1.13 (m, 5H). 13 C NMR (101 MHz, DMSO-d6) δ 166.60, 158.58, 154.92, 146.01, 140.17, 134.26, 130.95, 130.09, 128.41, 127.36, 126.79, 126.74, 105.87, 48.22, 33.39, 26.77, 25.70, 24.76.

[0122] Example 6: Synthesis of compound 6.

[0123]

[0124] The starting material p-aminocarbonylphenylboronic acid in Example 1 of the present application was replaced by 4-(N-methylcarboxamido)phenylboronic acid, and compound 5 was prepared by using similar experimental procedures to those in Example 1 of the present application.1 H NMR (400 MHz, DMSO-d6) δ 8.97 (s, 1H), 8.91 (s, 1H), 8.47 (s, 1H), 7.99 (s, 4H), 7.71 (s, 1H), 7.47 (s, 2H), 6.21 (s, 1H), 3.59 - 3.46 (m, 1H), 1.94 - 1.49 (m, 5H), 1.39 - 1.08 (m, 5H). 13 CNMR (101 MHz, DMSO-d6) δ 158.92, 154.80, 146.16, 144.02, 141.08, 130.57, 129.92, 128.13, 128.07, 126.99, 126.73, 106.55, 48.35, 33.41, 25.68, 24.84.

[0125] Example 7: Synthesis of compound 7.

[0126]

[0127] The starting material p-aminocarbonylphenylboronic acid in Example 1 of the present application was replaced with 1-isatyl-5-boronic acid pinacol ester, and compound 7 was prepared by using similar experimental procedures as in Example 1 of the present application. 1 H NMR (400 MHz, DMSO-d6) δ 8.97 (s, 1H), 8.91 (s, 1H), 8.47 (s, 1H), 7.99 (s, 4H), 7.71 (s, 1H), 7.47 (s, 2H), 6.21 (s, 1H), 3.59 - 3.46 (m, 1H), 1.94 - 1.49 (m, 5H), 1.39 - 1.08 (m, 5H). 13 CNMR (101 MHz, DMSO-d6) δ 170.08, 158.64, 154.79, 146.23, 145.67, 140.91, 132.71, 131.50, 129.90, 127.45, 126.79, 126.72, 124.05, 122.90, 106.32, 48.31, 45.50, 33.39, 25.67, 24.79.

[0128] Example 8: Synthesis of compound 8.

[0129]

[0130] The raw material p-carbamoylphenylboronic acid in the present application embodiment 1 is replaced with 2-methoxy-4-(4,4,5,5-tetramethyl-1,3,2-dioxaboran-2-yl)benzamide, and compound 8 is prepared by using similar experimental steps to the present application embodiment 1. 1 H NMR (400 MHz, DMSO-d6) δ 9.76 (s, 1H), 9.04 (s, 1H), 9.01 (s, 1H), 7.96 (d, J = 8.0 Hz, 1H), 7.78 - 7.57 (m, 4H), 7.52 (d, J = 8.0 Hz, 1H), 6.85 (d, J = 7.9 Hz, 1H), 4.08 (s, 3H), 3.63 - 3.48 (m, 1H), 1.88 - 1.48 (m, 5H), 1.41 - 1.15 (m, 5H). 13 C NMR (101 MHz, DMSO-d6) δ 166.43, 158.66, 158.37, 155.11, 145.95, 141.73, 132.11, 130.81, 130.43, 127.32, 126.90, 122.38, 119.23, 111.15, 105.10, 56.83, 48.01, 33.34, 25.77, 24.61.

[0131] Example 9: Synthesis of compound 9.

[0132]

[0133] The raw material p-carbamoylphenylboronic acid in the present application embodiment 1 is replaced with 3-chloro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaboran-2-yl)benzamide, and compound 9 is prepared by using similar experimental steps to the present application embodiment 1. 1 H NMR (400 MHz, DMSO-d6) δ 9.76 (s, 1H), 9.04 (s, 1H), 9.01 (s, 1H), 7.96 (d, J = 8.0 Hz, 1H), 7.78 - 7.57 (m, 4H), 7.52 (d, J = 8.0 Hz, 1H), 6.85 (d, J = 7.9 Hz, 1H), 4.08 (s, 3H), 3.63 - 3.48 (m, 1H), 1.88 - 1.48 (m, 5H), 1.41 - 1.15 (m, 5H). 13C NMR (101 MHz, DMSO-d6) δ 166.56, 158.62, 155.02, 147.31, 139.71, 136.08, 132.45, 132.35, 130.23, 129.83, 129.53, 129.31, 127.16, 126.27, 105.67, 48.07, 33.34, 25.72, 24.67.

[0134] Example 10: Synthesis of compound 10.

[0135]

[0136] The starting material p-carbamoylphenylboronic acid in Example 1 of the present application was replaced by 5-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-1 H- tetrazole, and compound 10 was prepared by using similar experimental procedures to those in Example 1 of the present application. 1 H NMR (400 MHz, DMSO-d6) δ 9.04 (s, 1 H), 8.99 (s, 1 H), 8.57 (s, 1 H), 8.48 (s, 1 H), 8.12 (d, J = 7.2 Hz, 1 H), 8.04 (d, J = 7.3 Hz, 1 H), 7.86 - 7.76 (m, 1 H), 7.72 (s, 1 H), 6.29 (s, 1 H), 4.04 - 3.93 (m, 1 H), 1.95 - 1.50 (m, 5H), 1.42 - 1.10 (m, 5H). 13 C NMR (101 MHz, DMSO-d6) δ 166.56, 158.62, 155.02, 147.31, 139.71, 136.08, 132.45, 132.35, 130.23, 129.83, 129.53, 129.31, 127.16, 126.27, 105.67, 48.07, 33.34, 25.72, 24.67.

[0137] Example 11 : Synthesis of compound 11.

[0138]

[0139] The starting material p-carbamoylphenylboronic acid in Example 1 of the present application was replaced by 4-methylsulfonamidophenylboronic acid, and compound 11 was prepared by using similar experimental procedures to those in Example 1 of the present application. 1H NMR (400 MHz, DMSO-d6) δ 9.96 (s, 1H), 8.96 (s, 1H), 8.87 (d, J = 2.1 Hz, 1H), 8.43 (d, J = 2.2 Hz, 1H), 7.80 (s, 1H), 7.77 (s, 1H), 7.68 (s, 1H), 7.39 (s, 1H), 7.37 (s, 1H), 6.27 (d, J = 7.8 Hz, 1H), 3.60 - 3.49 (m, 1H), 3.06 (s, 3H), 1.93 - 1.52 (m, 5H), 1.41 - 1.18 (m, 5H). 13 C NMR (101 MHz, DMSO-d6) δ 157.88, 155.60, 154.83, 145.89, 138.94, 133.10, 131.43, 129.88, 128.55, 126.75, 126.00, 120.54, 106.07, 48.29, 33.41, 25.68, 24.81.

[0140] Example 12: Synthesis of compound 12.

[0141]

[0142] The starting material p-carbamoylphenylboronic acid in Example 1 of the present application was replaced by 2-acetamidopyridine-5-boronic acid, and compound 12 was prepared by using similar experimental procedures to those in Example 1 of the present application. 1 H NMR (400 MHz, DMSO-d6) δ 10.71 (s, 1H), 8.93 (s, 1H), 8.87 (s, 1H), 8.75 (s, 1H), 8.42 (s, 1H), 8.31 - 8.16 (m, 2H), 7.70 (s, 1H), 6.19 (d, J = 7.6 Hz, 1H), 3.57 - 3.50 (m, 1H), 2.14 (s, 3H), 1.94 - 1.52 (m, 5H), 1.39 - 1.18 (m, 5H). 13 C NMR (101 MHz, DMSO-d6) δ 169.96, 158.25, 154.78, 152.37, 146.58, 145.78, 137.08, 129.81, 128.84, 128.80, 126.71, 125.93, 113.75, 106.29, 48.35, 33.41, 25.67, 24.84, 24.42.

[0143] Example 13: Synthesis of compound 13.

[0144]

[0145] The raw material p-aminocarbonylphenylboronic acid in Example 1 of the present application was replaced with p-propionamidophenylboronic acid, and compound 13 was prepared by using similar experimental procedures to those in Example 1 of the present application. 1 H NMR (400 MHz, DMSO-d6) δ 10.07 (s, 1H), 9.20 (s, 1H), 8.88 (s, 1H), 8.57 (s, 1H), 7.77 (s, 4H), 7.66 (s, 1H), 6.46 (d, J = 7.6 Hz, 1H), 3.60 - 3.49 (m, 1H), 2.37 (q, J = 7.4 Hz, 2H), 1.93 - 1.50 (m, 5H), 1.41 - 1.17 (m, 5H), 1.11 (t, J = 7.5 Hz, 3H). 13 C NMR (101 MHz, DMSO-d6) δ 172.66, 157.63, 154.93, 145.76, 139.90, 131.98, 131.56, 130.00, 127.78, 126.80, 125.91, 119.96, 105.60, 48.20, 46.07, 33.39, 30.03, 25.71, 24.75, 10.09, 9.02.

[0146] Example 14: Synthesis of compound 14.

[0147]

[0148] The raw material p-aminocarbonylphenylboronic acid in Example 1 of the present application was replaced with (1-acetylinindolin-5-yl)boronic acid, and compound 14 was prepared by using similar experimental procedures to those in Example 1 of the present application. 1 H NMR (400 MHz, DMSO-d6) δ 10.07 (s, 1H), 9.20 (s, 1H), 8.88 (s, 1H), 8.57 (s, 1H), 7.77 (s, 4H), 7.66 (s, 1H), 6.46 (d, J = 7.6 Hz, 1H), 3.60 - 3.49 (m, 1H), 2.37 (q, J = 7.4 Hz, 2H), 1.93 - 1.50 (m, 5H), 1.41 - 1.17 (m, 5H), 1.11 (t, J = 7.5 Hz, 3H). 13C NMR (101 MHz, DMSO-d6) δ 169.21, 157.60, 154.85, 145.85, 143.45, 133.55, 132.47, 131.82, 129.89, 126.73, 126.47, 125.73, 123.87, 116.64, 105.75, 48.90, 48.26, 33.40, 27.85, 25.70, 24.78, 24.48.

[0149] Example 15: Synthesis of compound 15.

[0150]

[0151] The starting material carbamoylphenylboronic acid in Example 1 of the present application was replaced by 3-ethoxycarbonylphenylboronic acid, and compound 15 was prepared by using similar experimental procedures to those in Example 1 of the present application. 1 H NMR (400 MHz, DMSO-d6) δ 8.93 (s, 2H), 8.44 (s, 1H), 8.31 (s, 1H), 8.09 (d, J = 7.7 Hz, 1H), 8.04 (d, J = 7.7 Hz, 1H), 7.76 - 7.63 (m, 2H), 6.20 (d, J = 7.7 Hz, 1H), 4.38 (q, J = 7.0 Hz, 2H), 3.58 - 3.49 (m, 1H), 1.92 - 1.49 (m, 5H), 1.42 - 1.14 (m, 8H). 13 C NMR (101 MHz, DMSO-d6) δ 166.03, 158.61, 154.82, 146.08, 138.43, 132.47, 131.37, 131.04, 130.25, 129.90, 129.12, 128.01, 126.70, 126.57, 106.37, 61.53, 48.37, 33.40, 25.67, 24.84, 14.67.

[0152] Example 16: Synthesis of compound 16.

[0153]

[0154] Into a three-necked flask, IV-2 (300 mg, 0.850 mmol), p-aminobenzamide (116 mg, 0.850 mmol), tris(dibenzylideneacetone)dipalladium (39 mg, 0.043 mmol), 2-dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl (20 mg, 0.043 mmol), cesium carbonate (831 mg, 2.550 mmol), t-butanol (20 mL) were added successively, and the system was heated at 100 °C for 12 hours. The reaction was monitored by TLC until completion. The reaction system was added with appropriate amount of water and ethyl acetate for extraction. The organic phase was retained, dried over anhydrous sodium sulfate, and then the organic phase was mixed with silica gel and separated and purified by column chromatography (DCM:MeOH, v:v = 35:1) to obtain the white solid target compound 16 (45 mg). 1 H NMR (400 MHz, DMSO-d6) δ 8.73 (s, 1H), 8.58 (s, 1H), 8.37 (s, 1H), 8.00 - 7.90 (m, 2H), 7.62 (s, 1H), 7.59 (s, 1H), 7.41 - 7.23 (m, 4H), 6.27 (d, J = 7.7 Hz, 1H), 3.55 - 3.44 (m, 1H), 1.89 - 1.48 (m, 5H), 1.38 - 1.10 (m, 5H). 13 C NMR (101 MHz, DMSO-d6) δ 168.51, 154.94, 151.12, 144.06, 141.17, 136.77, 136.23, 129.80, 129.43, 127.07, 119.58, 119.26, 115.84, 115.51, 106.71, 48.32, 33.43, 25.68, 24.85.

[0155] Example 17: Synthesis of compound 17.

[0156]

[0157] The starting material p-aminobenzamide in Example 16 of the present application was replaced with methyl 4-aminobenzoate, and compound 17 was prepared by using similar experimental procedures to those in Example 16 of the present application. 1H NMR (400 MHz, DMSO-d6) δ 9.04 (s, 1H), 8.68 (s, 1H), 8.41 (s, 1H), 8.06 (s, 1H), 7.87 (s, 1H), 7.85 (s, 1H), 7.65 (s, 1H), 7.14 (s, 1H), 7.12 (s, 1H), 6.15 (d, J = 7.7 Hz, 1H), 3.80 (s, 3H), 3.58 - 3.44 (m, 1H), 1.91 - 1.50 (m, 5H), 1.38 - 1.09 (m, 5H). 13 C NMR (101 MHz, DMSO-d6) δ 166.44, 154.82, 152.67, 149.02, 142.24, 134.96, 131.72, 129.41, 126.96, 120.22, 118.00, 114.59, 106.97, 52.06, 48.38, 33.42, 25.66, 24.89.

[0158] Example 18: Synthesis of compound 18.

[0159]

[0160] The raw material cyclohexylamine in Example 1 of the present application was replaced by 4-aminotetrahydropyran, and compound 18 was prepared by using similar experimental procedures to Example 1 of the present application. 1 H NMR (400 MHz, DMSO-d6) δ 9.43 (s, 1H), 8.97 (s, 1H), 8.77 (s, 1H), 8.16 - 7.88 (m, 5H), 7.70 (s, 1H), 7.45 (s, 1H), 6.71 (d, J = 7.0 Hz, 1H), 3.92 - 3.70 (m, 3H), 3.51 - 3.36 (m, 2H), 1.90 - 1.32 (m, 4H). 13 C NMR (101 MHz, DMSO-d6) δ 167.89, 158.59, 155.04, 145.99, 140.33, 134.06, 130.92, 130.09, 128.85, 127.24, 126.90, 126.85, 105.97, 66.19, 45.83, 33.58.

[0161] Example 19: Synthesis of compound 19.

[0162]

[0163] The raw material cyclohexylamine in Example 1 of the present application was replaced by cyclopropylamine, and compound 19 was prepared by using similar experimental procedures to Example 1 of the present application. 1H NMR (400 MHz, DMSO-d6) δ 8.97 (s, 1H), 8.90 (s, 1H), 8.47 (s, 1H), 8.18 - 8.01 (m, 3H), 7.96 - 7.86 (m, 2H), 7.71 (s, 1H), 7.48 (s, 1H), 6.24 (d, J = 4.9 Hz, 1H), 3.80 - 3.67 (m, 1H), 1.96 - 1.82 (m, 2H), 1.66 - 1.34 (m, 10H). 13 C NMR (101 MHz, DMSO-d6) δ 167.89, 158.62, 154.69, 146.12, 140.43, 134.12, 131.06, 129.92, 128.87, 127.36, 126.70, 126.47, 106.16, 50.53, 35.28, 28.18, 23.96.

[0164] Example 20: Synthesis of compound 20.

[0165]

[0166] The raw material cyclohexylamine in Example 1 of the present application was replaced by cycloheptylamine, and compound 20 was prepared by using similar experimental procedures to Example 1 of the present application. 1 H NMR (400 MHz, DMSO-d6) δ 8.97 (s, 1H), 8.90 (s, 1H), 8.47 (s, 1H), 8.18 - 8.01 (m, 3H), 7.96 - 7.86 (m, 2H), 7.71 (s, 1H), 7.48 (s, 1H), 6.24 (d, J = 4.9 Hz, 1H), 3.80 - 3.67 (m, 1H), 1.96 - 1.82 (m, 2H), 1.66 - 1.34 (m, 10H). 13 C NMR (101 MHz, DMSO-d6) δ 167.89, 158.62, 154.69, 146.12, 140.43, 134.12, 131.06, 129.92, 128.87, 127.36, 126.70, 126.47, 106.16, 50.53, 35.28, 28.18, 23.96.

[0167] Example 21: Synthesis of compound 21.

[0168]

[0169] The raw material cyclohexylamine in Example 1 of the present application was replaced by aniline, and compound 21 was prepared by using similar experimental procedures to Example 1 of the present application. 1H NMR (400 MHz, DMSO-d6) δ 9.28 (s, 1H), 9.00 (s, 1H), 8.79 (s, 1H), 8.53 (s, 1H), 8.21 - 7.90 (m, 5H), 7.85 (s, 1H), 7.61 - 7.27 (m, 5H), 7.09 - 6.97 (m, 1H). 13 C NMR (101 MHz, DMSO-d6) δ 167.91, 158.60, 152.98, 146.31, 140.39, 139.87, 134.17, 131.19, 129.39, 129.16, 128.90, 127.40, 126.84, 126.68, 122.60, 118.68, 108.37.

[0170] Example 22: Synthesis of compound 22.

[0171]

[0172] The starting material cyclohexylamine in Example 1 of the present application was replaced by benzylamine, and compound 22 was prepared by using similar experimental procedures to those in Example 1 of the present application. 1 H NMR (400 MHz, DMSO-d6) δ 9.28 (s, 1H), 9.00 (s, 1H), 8.79 (s, 1H), 8.53 (s, 1H), 8.21 - 7.90 (m, 5H), 7.85 (s, 1H), 7.61 - 7.27 (m, 5H), 7.09 - 6.97 (m, 1H). 13 C NMR (101 MHz, DMSO-d6) δ 167.91, 158.60, 152.98, 146.31, 140.39, 139.87, 134.17, 131.19, 129.39, 129.16, 128.90, 127.40, 126.84, 126.68, 122.60, 118.68, 108.37.

[0173] Example 23: Synthesis of compound 23.

[0174]

[0175] The starting material cyclohexylamine in Example 1 of the present application was replaced by trans-4-aminocyclohexanol, and compound 23 was prepared by using similar experimental procedures to those in Example 1 of the present application. 1H NMR (400 MHz, DMSO-d6) δ 8.96 (s, 1H), 8.90 (s, 1H), 8.46 (s, 1H), 8.14 - 7.84 (m, 5H), 7.70 (s, 1H), 7.47 (s, 1H), 6.13 (d, J = 7.6 Hz, 1H), 4.56 (d, J = 4.1 Hz, 1H), 3.57 - 3.39 (m, 2H), 2.02 - 1.03 (m, 8H). 13 C NMR (101 MHz, DMSO-d6) δ 167.91, 158.62, 154.95, 146.10, 140.41, 134.11, 131.04, 129.89, 128.87, 127.33, 126.74, 126.55, 106.36, 68.45, 48.33, 34.25, 31.23.

[0176] Example 24: Synthesis of compound 24.

[0177]

[0178] The raw material phenyl chloroformate in Example 1 of the present application was replaced by phenyl thiocarbamates, and compound 24 was prepared by using similar experimental procedures in Example 1 of the present application. 1 H NMR (400 MHz, DMSO-d6) δ 9.59 (s, 1H), 8.97 (s, 1H), 8.42 (s, 1H), 8.26 (s, 1H), 8.15 - 7.82 (m, 6H), 7.46 (s, 1H), 4.21 - 4.03 (m, 1H), 2.05 - 1.53 (m, 5H), 1.43 - 1.06 (m, 5H). 13 C NMR (101 MHz, DMSO-d6) δ 179.90, 167.90, 158.52, 146.26, 140.37, 134.14, 131.26, 129.90, 128.88, 128.45, 127.42, 115.21, 52.99, 32.33, 25.61, 25.04.

[0179] Example 25: Synthesis of compound 25.

[0180]

[0181] The raw material p-carbamoylphenylboronic acid in Example 1 of the present application was replaced by p-acetamidophenylboronic acid, and compound 25 was prepared by using similar experimental procedures in Example 1 of the present application. 1HNMR (400 MHz, DMSO-d6) δ 10.14 (s, 1H), 9.06 (s, 1H), 8.87 (s, 1H), 8.50 (s, 1H), 7.76 (s, 4H), 7.67 (s, 1H), 6.45 (d, J = 7.0 Hz, 1H), 4.06 - 3.90 (m, 1H), 2.09 (s, 3H), 1.94 - 1.82 (m, 2H), 1.74 - 1.38 (m, 6H). 13 C NMR (101 MHz, DMSO-d6) δ 168.98, 157.66, 155.25, 145.81, 139.85, 132.12, 131.60, 129.90, 127.83, 126.77, 125.85, 119.93, 105.83, 51.49, 33.33, 24.55, 23.62.

[0182] Example 26: Synthesis of compound 26.

[0183]

[0184] The raw material p-aminocarbonylphenylboronic acid in Example 1 of the present application was replaced with p-acetylamino phenylboronic acid, and cyclohexylmethylamine was replaced with cyclohexylmethylamine, and compound 26 was prepared by using similar experimental procedures to Example 1 of the present application. 1 H NMR (400 MHz, DMSO-d6) δ 10.13 (s, 1H), 9.03 (s, 1H), 8.87 (s, 1H), 8.45 (s, 1H), 7.75 (s, 4H), 7.67 (s, 1H), 6.38 (s, 1H), 3.07 - 2.97 (m, 2H), 2.09 (s, 3H), 1.87 - 1.56 (m, 6H), 1.49 - 1.38 (m, 1H), 1.31 - 1.04 (m, 4H). 13 C NMR (101 MHz, DMSO-d6) δ 168.98, 157.68, 155.72, 145.85, 139.85, 132.15, 131.64, 129.92, 127.87, 126.78, 125.86, 119.93, 106.13, 45.84, 38.45, 30.80, 26.54, 25.88, 24.55.

[0185] Example 27: Evaluation of DRAK2 inhibitory activity of some compounds of the present application.

[0186] 1. Materials:

[0187] Some compounds of the present application (see Table 1 in particular).

[0188] 2. Test principle:

[0189] DRAK2 is a serine / threonine protein kinase. ADP-Glo TM Kinase Assay (Promega; v9102) is a luminescence-based kinase assay kit that detects ADP formed in a kinase reaction. The kit converts ADP generated in the reaction to ATP, which is then converted to light by Ultra-Glo TM Luciferase. The emitted light signal is directly proportional to the amount of ATP present, and this method can be used to determine the inhibitory activity of small molecule compounds against DRAK2.

[0190] 3. Test method:

[0191] Before the test, some of the compounds described in the present application are dissolved in DMSO to prepare a stock solution, which is diluted to the required concentration with culture solution before use. 1 μL of the compound is added to each well for each determination, and 2 μL of the kinase solution is added to each well of the assay plate except for the control wells without enzyme. 2 μL of ATP is added to each well of the assay plate, the plate is shaken and centrifuged. The assay is stopped by adding 2.5 μL of ADP-Glo TM reagent to terminate the kinase reaction and consume the unspent ATP, leaving only ADP and very low ATP background. Detection analysis, 5 μL of kinase detection reagent is added to convert ADP to ATP and introduce luciferase and luciferin to detect ATP. The luminescence is measured using Envision, the values are copied from the Envision program, and the data are processed using GraphPad Prism 5.0. The formula for calculating the percentage of activity is Activity% = (sample Ratio - minimum value) / (maximum value - minimum value) x 100 (the maximum value refers to the value of the control group with 2% DMSO; the minimum value refers to the value of the control group without enzyme). In this way, the inhibitory activity data of small molecule compounds against DRAK2 can be obtained.

[0192] 4. Results and discussion:

[0193] Table 1: Test results of DRAK2 inhibitory activity of some compounds of the present application (including but not limited to these compounds)

[0194]

[0195]

[0196] Note: IC 50 is the evaluation of the inhibitory activity of the sample drug against DRAK2, and the half inhibitory concentration.

[0197] The test results show that some of the compounds of the present application, such as 1-8, 10-16, 18-26, can inhibit DRAK2 kinase activity at a level less than 1000 nM. Among them, compounds 1, 2, 5, 6, 7, 8, 11, 13, 14, 20, 25 can inhibit DRAK2 kinase activity at a level less than 100 nM. Such compounds as a novel DRAK2 inhibitor can be used for the development of new therapeutic drugs for metabolic syndrome including but not limited to diabetes, non-alcoholic fatty liver disease, etc.

[0198] Example 28: Evaluation of the function of some of the compounds of the present application on mouse islet glucose-stimulated insulin release (GSIS).

[0199] 1. Materials:

[0200] Some of the compounds of the present application (see Table 2 in particular).

[0201] 2. Test principle:

[0202] Mouse primary islets have strong insulin releasing ability and can release insulin in response to glucose stimulation under high glucose (16.7 mM glucose) stimulation. The insulin content in the culture medium is detected to evaluate the effect of the compounds on islet function. Mouse primary islets are obtained by mouse islet isolation technology, and are co-treated with high and low glucose (2.8 mM glucose) respectively by some of the compounds of the present application. The present application detects the insulin content in the supernatant to evaluate whether some of the compounds of the present application have a promoting or protective effect on the insulin releasing ability. HTRF is based on two technologies of fluorescence resonance energy transfer and time-resolved fluorescence to detect the concentration of the measured insulin in the collected cell supernatant.

[0203] 3. Test method:

[0204] Part of the compounds described in the application are dissolved in DMSO, stored at low temperature, and the concentration of DMSO in the final system is controlled within the range that does not affect the detection activity. Well-grown, uniform, and uniform primary mouse islets are inoculated into a 48-well plate at a density of 10 per well. The mouse primary islets are starved for 1 hour with KRBH buffer containing 0.1% BSA (without free fatty acids). The supernatant is discarded, and KRBH buffer containing 0.1% BSA and 2.8 mM glucose and the corresponding group of compounds described in the application are added for stimulation under low-sugar conditions for 1 hour, and the supernatant is collected for detection. Then, 0.1% BSA and 16.7 mM glucose KRBH buffer and the corresponding group of compounds described in the application are added to the corresponding plate wells for stimulation under high-sugar conditions for 1 hour, and the supernatant is collected for detection. The insulin content in the collected supernatant is detected using the Insulin HTRF detection kit (PerkinElmer; #62INSPEC), and the EnVisionTM signal intensity is detected. The concentration (ng / mL) of each sample is calculated by establishing a standard curve based on the readings of each compound well, and then the actual insulin content (ng / μg) of each group is calculated based on the protein amount of each group of islets, and then the fold value can be calculated.

[0205] 4. Results and discussion:

[0206] Table 2: Functional evaluation results of part of the compounds of the application (including but not limited to these compounds)

[0207] Compound GSIS (fold) 1 2.3 2 2.3 3 4 2.67 5 3.24 6 1.39 8 1.12 22 1.86 23 4.11 22b 1.41

[0208] Note: fold is the fold of the insulin secretion amount of the experimental group relative to the blank control group after high-sugar treatment at a concentration of 5 μM of the compound, and 22b is a positive control compound.

[0209] The evaluation results show that compounds 4, 5, 6, 8, 22, and 23 can effectively promote the secretion of insulin by mouse islet cells, and the insulin secretion promoting activity of compounds 4, 5, 22, and 23 is significantly better than that of the reported DRAK2 small molecule inhibitor 22b. Such compounds exhibit excellent insulin secretion promoting activity and can be used for the development of new therapeutic drugs for metabolic syndromes including but not limited to diabetes.

[0210] Unless otherwise defined, all technical and scientific terms used in the present application have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. The terminology used in the description of the application is for the purpose of describing particular embodiments only and is not intended to be limiting of the application.

[0211] As used in the present application, the terms "comprise", "comprising", are open-ended expressions, i.e. they include the indicated content but not exclude other aspects.

[0212] As used herein, the term "and / or" includes any and all combinations of one or more of the associated listed items.

[0213] All documents referred to in this application are incorporated herein by reference as if each individual document were incorporated by reference. In addition, it is to be understood that the application can be carried out by specifically different embodiments and that embodiments can be practiced without materials details, quantifications, and other specific items, parts, and pieces specifically recited herein. These equivalents are also within the scope of the application. Also, it is to be understood that the application is not limited to the particular methodology, protocols, and materials described herein and as such can involve any procedures or agents that produce the same or similar results.

Claims

1. A 3-ureidosubstituted thiopheno[2,3-b]pyridine compound or a pharmaceutically acceptable salt thereof, characterized in that, The structure of the compound is shown in formula (I): in, Ring A is selected from the following group: phenyl, pyridyl; R1 is selected from the following groups: -C(O)ORa, -NHS(O)2Rb, -S(O)2NH2, -C(O)N(Ra)2, -NHC(O)Rb. In this context, Ra is independently selected from the group consisting of hydrogen and C1-C6 alkyl; Rb is selected from the group consisting of C1-C6 alkyl; R2 is selected from the group consisting of hydrogen, halogen, C1-C6 alkoxy, and C1-C6 alkyl; or, R1 and R2 located on adjacent ring atoms of ring A together with their adjacent ring atoms constitute a 5-membered heterocyclic group. R3 is selected from the following group: cyclopropyl, cyclopentyl, cyclohexyl, cycloheptyl, phenyl, benzyl. R4 is selected from hydrogen; L is selected from the following group: covalent single bond, -NH-; X is selected from the following group: oxygen, sulfur.

2. The compound of claim 1 or a pharmaceutically acceptable salt thereof, characterized in that, The ring A is selected from the group consisting of: phenyl, pyridyl; and / or, The R1 is selected from the following group: -C(O)ORa, -NHS(O)2Rb, -S(O)2NH2, -C(O)N(Ra)2, -NHC(O)Rb, Wherein, Ra is independently selected from the group consisting of hydrogen, methyl, ethyl, and propyl; and / or, Rb is selected from the group consisting of methyl and ethyl; and / or, The R2 is selected from the group consisting of: hydrogen, fluorine, chlorine, methoxy; and / or, The R3 is selected from the group consisting of: cyclopropyl, cyclopentyl, cyclohexyl, cycloheptyl, phenyl, benzyl, And / or, The R4 is selected from hydrogen; and / or, The L is selected from a covalent single bond; and / or, X is selected from oxygen.

3. The compound of claim 1 or a pharmaceutically acceptable salt thereof, characterized in that, The compound of formula (I) has the structure shown in formula (III-1) or (III-2): The definitions of R1, R2, and R3 are the same as in claim 1.

4. The compound of claim 1 or a pharmaceutically acceptable salt thereof, characterized in that, The compounds of formula (I) are selected from the following group:

5. A method for preparing a 3-ureidosubstituted thiopheno[2,3-b]pyridine compound, characterized in that, The preparation method includes the following route: (1) In a first solvent, in the presence of a base, 5-bromothiopheno[2,3-b]pyridine-3-amine undergoes a nucleophilic substitution reaction with phenyl chloroformate or phenyl thiochloroformate to generate intermediate IV-1; the first solvent is one or more of dichloromethane, tetrahydrofuran, and dimethyl sulfoxide; the base is one or two of potassium carbonate and sodium bicarbonate. (2) In a second solvent, in the presence of a base, the intermediate IV-1 obtained in step (1) undergoes a nucleophilic substitution reaction with a substituted amine to generate intermediate IV-2; the second solvent is one or more of dimethyl sulfoxide, acetonitrile, and toluene; the base is one or two of potassium carbonate and triethylamine. (3) In a third solvent, in the presence of a base and a palladium catalyst, the intermediate IV-2 obtained in step (2) undergoes a palladium-catalyzed Suzuki coupling reaction with a substituted arylboronic acid or borate ester under alkaline conditions to generate the target compound II-1; the third solvent is one or more of dimethyl sulfoxide, N,N-dimethylformamide, toluene, and water; the palladium catalyst is one or more of tetratetraphenylphosphine palladium, bis(triphenylphosphine)palladium dichloride, and 1,1'-bis(diphenylphosphine)ferrocene palladium dichloride dichloromethane complex; the base is one or two of potassium carbonate and sodium carbonate. or, (4) In a fourth solvent, in the presence of a base, a palladium catalyst, and a phosphine ligand, the intermediate IV-2 obtained in step (2) undergoes a palladium-catalyzed Buchwald–Hartwig coupling reaction with a substituted aromatic amine under alkaline conditions to generate the target compound II-2; the fourth solvent is one or more of toluene, dioxane, and tert-butanol; the palladium catalyst is one or more of tris(dibenzylacetone)dipalladium, tetratriphenylphosphine palladium, and bis(triphenylphosphine)dichloride palladium; the phosphine ligand is one or more of (S)-(-)-1,1′-binaphthyl-2,2′-bis(diphenylphosphine), 4,5-bis(diphenylphosphino)-9,9-dimethyloxanthracene, and 2-dicyclohexylphosphine-2′,4′,6′-triisopropylbiphenyl; the base is one or two of cesium carbonate and potassium tert-butoxide. The reaction process along the route is shown in reaction formula (a): Wherein, R5 is B(OH)2 or B(Pin); the definitions of rings A, R1, R2, R3, R4, and X are the same as in claim 1.

6. The preparation method according to claim 5, characterized in that, In step (1), the ratio of the first solvent to 5-bromothieno[2,3-b]pyridine-3-amine is 1 mL:0.02 mmol to 1 mL:0.04 mmol; and / or, the molar ratio of 5-bromothieno[2,3-b]pyridine-3-amine to phenyl chloroformate or phenyl thiochloroformate is 1:1 to 1:1.5; and / or, the molar ratio of the base to 5-bromothieno[2,3-b]pyridine-3-amine is 1:1 to 3:1; and / or, the reaction temperature is 20°C to 40°C; and / or, the reaction time is 1 to 2 hours. And / or, In step (2), the ratio of the second solvent to intermediate IV-1 is 1 mL:0.02 mmol to 1 mL:0.04 mmol; and / or, the substituted amine is one or more of cyclopentylamine, cyclohexylamine, cycloheptylamine, 4-aminotetrahydropyran, cyclopropylamine, aniline, benzylamine, trans-4-aminocyclohexanol, and cyclohexylmethylamine; and / or, the molar ratio of intermediate IV-1 to the substituted amine is 1:1 to 1:1.5; and / or, the molar ratio of the base to intermediate IV-1 is 1:1 to 3:1; and / or, the reaction temperature is 20°C to 40°C; and / or, the reaction time is 1 to 2 hours. And / or, In step (3), the ratio of the third solvent to intermediate IV-2 is 1 mL:0.01 mmol to 1 mL:0.03 mmol; and / or, the substituted arylboronic acid or borate ester is p-carbamoylphenylboronic acid, p-carboxyphenylboronic acid, 1-isoindolone-5-boronic acid pinacol ester, 5-carboxy-2-fluorophenylboronic acid, p-acetaminophenylboronic acid, 4-(N-methylformamide)phenylboronic acid, 4-(aminosulfonyl)phenylboronic acid, 2-methoxy-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)benzamide, 3-chloro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)benzamide, 5-(3-(4,4,5,5-tetramethyl-1,3,2-) One or more of the following: dioxoborane-2-yl)phenyl)-1H-tetrazole, 4-methylsulfonamide phenylboronic acid, 2-acetamidopyridine-5-boronic acid, p-propamidophenylboronic acid, (1-acetylindoline-5-yl)boronic acid, and 3-ethoxycarbonylphenylboronic acid; and / or, the molar ratio of intermediate IV-2 to the substituted arylboronic acid or boronic ester is 1:1 to 1:5; and / or, the molar ratio of intermediate IV-2 to the palladium catalyst is 1:0.05 to 1:0.10; and / or, the molar ratio of the base to intermediate IV-2 is 2:1 to 3:1; and / or, the reaction temperature of the Suzuki coupling reaction is 80°C to 100°C; and / or, the reaction time of the Suzuki coupling reaction is 5 to 12 hours. or, In step (4), the ratio of the fourth solvent to intermediate IV-2 is 1 mL:0.03 mmol to 1 mL:0.05 mmol; and / or, the substituted aromatic amine includes one or more of p-aminobenzoamide and methyl 4-aminobenzoate; and / or, the molar ratio of intermediate IV-2 to the substituted aromatic amine is 1:1 to 1:5; and / or, the molar ratio of intermediate IV-2 to the palladium catalyst is 1:0.

05. ~1:0.10; and / or, the molar ratio of the intermediate IV-2 to the phosphine ligand is 1:0.05 to 1:0.10; and / or, the molar ratio of the base to the intermediate IV-2 is 2:1 to 3:1; and / or, the reaction temperature of the Buchwald–Hartwig coupling reaction is 80°C to 100°C; and / or, the reaction time of the Buchwald–Hartwig coupling reaction is 5 to 12 hours.

7. A drug / drug composition, characterized in that, It comprises any one of the compounds of claims 1-4 or a pharmaceutically acceptable salt thereof; and a pharmaceutically acceptable carrier.

8. The use of any compound of claims 1-4 or a pharmaceutically acceptable salt thereof, or the preparation method of claim 5 or 6, or the drug / drug composition of claim 7 in the preparation of a medicament for treating or preventing diseases related to DRAK2 activity or expression, or a medicament for inhibiting DRAK2 kinase activity.

9. The application as described in claim 8, characterized in that, The disease is metabolic syndrome; wherein, the metabolic syndrome is diabetes.

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