3-ureido substituted thieno [2, 3-b] pyridine compound as well as preparation method and application thereof

Abstract

The invention discloses a 3-ureido substituted thieno [2, 3-b] pyridine compound with a novel structure as well as a preparation method and application of the 3-ureido substituted thieno [2, 3-b] pyridine compound. The preparation method comprises the following steps: carrying out nucleophilic substitution reaction on 5-bromothieno [2, 3-b] pyridine-3-amine and phenyl chloroformate or phenyl thiochloroformate to generate an intermediate, and carrying out nucleophilic substitution reaction on the intermediate and substituted amine to generate a key intermediate; then, the key intermediate and substituted arylboronic acid or boric acid ester or substituted arylamine are subjected to a palladium catalyzed Suzuki coupling reaction or a Buchwald-Harwig coupling reaction under the alkaline condition, and the target product 3-ureido substituted thieno [2, 3-b] pyridine compound is obtained. The 3-ureido substituted thieno [2, 3-b] pyridine compound disclosed by the invention, as a DRAK2 inhibitor, has a certain prospect in the development of drugs for treating metabolic syndromes.

Description

Technical Field

[0001] The invention 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 Art

[0002] DRAK2 (death-related 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 the thymus, spleen, and lymph nodes, and is also expressed to a certain extent in other tissues and organs such as the olfactory lobe, ventricular zone, hippocampus, intestinal epithelial cells, and pancreas of the brain (J.Immunol.2009,182,4762-4770; Front.Pharmacol.2022,13,1014508). Recent studies have found that DRAK2 protein is abnormally highly expressed in pancreatic tissues of hyperglycemic humans and diabetic monkeys. Increased DRAK2 protein levels, blocked autophagy, and increased apoptosis were also detected in the pancreas of diabetic model mice (db / db mice). By constructing DRAK2-cKO mice with specific knockout of DRAK2 in pancreatic β cells, it was found that DRAK2-cKO mice can resist islet function damage induced by a high-fat diet. Other studies have found that DRAK2 expression is significantly increased in liver biopsy tissues of NAFLD / NASH model mice and patients, and is positively correlated with NASH pathological scores.

[0003] Molecular mechanism studies have shown that in pancreatic β cells, DRAK2 has a direct protein-protein interaction with ULK1, a key protein of the autophagy initiation complex, and directly phosphorylates the ULK1-Ser56 site, thereby promoting the ubiquitination degradation of ULK1, thereby negatively regulating autophagy, damaging mitochondrial quality and β cell function (Sci. Transl. Med. 2024, 16, eade8647). In liver cells, DRAK2 has a direct protein-protein interaction with the RNA splicing factor SRSF6, which leads to abnormal alternative splicing of mitochondrial function-related genes (including mtDNA polymerase POLγ2). This abnormal alternative splicing of mitochondrial function-related genes is a key factor in the occurrence and 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 very valuable to develop new structures of DRAK2 small molecule inhibitors, and it is particularly urgent to obtain small molecules with high inhibitory activity against DRAK2 and safe and effective. Summary of the invention

[0004] In order to solve the deficiencies of the prior art, the purpose of the present invention 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 to develop therapeutic drugs for metabolic syndrome, including but not limited to: diabetes, non-alcoholic fatty liver disease, hyperlipidemia, obesity, and biliary cirrhosis.

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

[0006]

[0007] in,

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

[0009] R1 is selected from the following group: -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 Ra is independently selected from the following group: hydrogen, cyclopropyl, C1-C6 alkyl; Rb is selected from the following group: substituted or unsubstituted C1-C6 alkyl, phenyl; R2 is selected from the following group: hydrogen, hydroxyl, amine, halogen, C1-C6 alkyl, C1-C6 alkoxy; or, R1 and R2 located on adjacent ring atoms of ring A may form together with their adjacent ring atoms: a substituted or unsubstituted 5-7 membered heterocyclyl;

[0010] R3 is selected from the group consisting of cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, phenyl, pyridyl, substituted or unsubstituted benzyl,

[0011] R4 is 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 otherwise specified, the substitution involved in the above groups means that the hydrogen atoms on the groups are replaced by one to three substituents selected from the following groups: halogen, hydroxyl, amine, C1-C3 alkyl, C1-C3 alkoxy, acetyl,

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

[0016] In another preferred embodiment, R1 is selected from the following group: -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, Ra is independently selected from the following group: hydrogen, methyl, ethyl, propyl, cyclopropyl, isopropyl, tert-butyl; Rb is selected from the following group: methyl, chloromethyl, ethyl, phenyl.

[0017] In another preferred embodiment, R2 is selected from the following group: hydrogen, hydroxy, fluorine, chlorine, methyl, methoxy.

[0018] In another preferred embodiment, R3 is selected from the group consisting of cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, phenyl, pyridyl, benzyl,

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

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

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

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

[0023]

[0024] Wherein, ring A, R1, R2, R3, R4, and X are defined the same as in formula (I).

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

[0026]

[0027] Wherein, R1, R2, and R3 are defined the same as in formula (I).

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

[0029]

[0030] The present invention also provides a method for preparing a 3-ureido-substituted thieno[2,3-b]pyridine compound (as shown in formula (I)), the preparation method comprising the following route:

[0031] (1) In the presence of a base, 5-bromothieno[2,3-b]pyridin-3-amine reacts with phenyl chloroformate or phenyl chlorothioate to form an intermediate IV-1 by a nucleophilic substitution reaction in a first solvent;

[0032] (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 an intermediate IV-2;

[0033] (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 aryl boronic acid or boric acid ester under alkaline conditions to generate the target compound II-1;

[0034] or,

[0035] 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;

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

[0037]

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

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

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

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

[0042] In the step (1), the base is one or more of potassium carbonate, sodium bicarbonate, etc.; preferably, it is 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 reaction temperature is 20°C to 40°C; preferably, 25°C.

[0045] In the step (1), the reaction time 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, it is dimethyl sulfoxide.

[0047] In the step (2), the ratio of the amount of the second solvent to the amount of the intermediate IV-1 (ml:mmol) is 1:0.02 to 1:0.04; preferably, it is 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, etc., 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; preferably, it is 1:1.5.

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

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

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

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

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

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

[0056] In the step (3), the substituted aromatic boronic acid or boronic acid ester includes p-aminoformylphenylboronic acid, p-carboxyphenylboronic acid, 1-isoindolone-5-boronic acid pinacol ester, 5-carboxy-2-fluorophenylboronic acid, p-acetylaminophenylboronic acid, 4-(N-methylformamido)phenylboronic acid, 4-(aminosulfonyl)phenylboronic 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-methylsulfonylaminophenylboric acid, 2-acetylaminopyridine-5-boric acid, p-propionamidophenylboric acid, (1-acetylindolin-5-yl)boric acid, 3-ethoxycarbonylphenylboric acid, etc., preferably, p-carbamoylphenylboric 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 tetrakistriphenylphosphine palladium, bistriphenylphosphine palladium (II) dichloride, 1,1'-bis(diphenylphosphino)ferrocene palladium (II) dichloromethane complex, etc.; preferably, it is tetrakistriphenylphosphine 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, it is 1:0.10.

[0060] In the step (3), the base is one or more of potassium carbonate, sodium carbonate, etc.; preferably, it is 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, it is 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, tert-butyl alcohol, etc.; preferably, tert-butyl alcohol. In the step (3), the ratio of the fourth solvent to the amount of intermediate IV-2 (ml:mmol) is 1:0.03 to 1:0.05; preferably, it is 1:0.04.

[0066] In the step (3), the substituted aromatic amine includes one or more of p-aminobenzamide, 4-aminobenzoic acid methyl ester, etc., preferably p-aminobenzamide.

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

[0068] In the step (3), the palladium catalyst is one or more of tris(dibenzylideneacetone)dipalladium, tetrakistriphenylphosphine palladium, bistriphenylphosphine palladium (II) dichloride, etc.; preferably, it is 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, it is 1:0.05.

[0070] In the step (3), the phosphine ligand is one or more of (S)-(-)-1,1′-binaphthyl-2,2′-bisdiphenylphosphine, 4,5-(bisdiphenylphosphino)-9,9-dimethylxanthene, 2-dicyclohexylphospho-2′,4′,6′-triisopropylbiphenyl, etc.; preferably, it is 2-dicyclohexylphospho-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, it is 1:0.05.

[0072] In the step (3), the base is one or more of cesium carbonate, potassium tert-butoxide, etc.; preferably, it is 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 a specific embodiment, the preparation method of the 3-ureido-substituted thieno[2,3-b]pyridine compound (as shown in formula (I)) comprises the following steps:

[0077] (1) Using 5-bromothieno[2,3-b]pyridin-3-amine as a raw material and phenyl chloroformate or phenyl chlorothioformate in a molar ratio of 1:1.5, sodium bicarbonate as a base, and tetrahydrofuran as a solvent, a nucleophilic substitution reaction is carried out at 25° C. for 1 to 2 hours to obtain intermediate IV-1;

[0078] (2) The molar ratio of the intermediate IV-1 to the substituted amine is 1:1.5, triethylamine is used as a base, dimethyl sulfoxide is used as a solvent, and a nucleophilic substitution reaction is carried out at 25° C. for 1 to 2 hours to obtain the intermediate IV-2;

[0079] (3) The molar ratio of the intermediate IV-2 to the substituted arylboronic acid or boronic acid ester is 1:1, potassium carbonate is used as a base, N,N-dimethylformamide and water are used as solvents, and a Suzuki coupling reaction catalyzed by tetrakistriphenylphosphine palladium (10 mol%) is carried out at 85° C. for 5 to 12 hours to produce the target compound II-1;

[0080] or,

[0081] The molar ratio of intermediate IV-2 and substituted aromatic amine is 1:1, cesium carbonate is used as a base, tert-butanol is used as a solvent, 2-dicyclohexylphosphine-2′,4′,6′-triisopropylbiphenyl is used as a phosphine ligand, and a Buchwald–Hartwig coupling reaction catalyzed by tri(dibenzylideneacetone)dipalladium (5 mol%) is carried out at 100°C. The reaction takes 5 to 12 hours to produce the target compound II-2.

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

[0083]

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

[0085] The present invention also provides a drug / drug composition comprising a compound as described above, such as formula (I), formula (II-1), formula (II-2), formula (III-1), formula (III-2) or a stereoisomer thereof, a pharmaceutically acceptable salt, a prodrug or a solvate; and a pharmaceutically acceptable carrier.

[0086] Furthermore, the drug / drug composition is used alone and / or in combination with other drugs.

[0087] Preferably, the pharmaceutically acceptable carrier refers to a drug that does not produce adverse, allergic or other adverse reactions when the drug is properly administered to animals or humans. Pharmaceutically acceptable carriers include, 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 methylcellulose, ethyl cellulose and methyl cellulose; tragacanth powder; 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; polyols, such as propylene glycol, glycerol, sorbitol, mannitol and polyethylene glycol; alginic acid; emulsifiers, such as Tween; wetting agents, such as sodium lauryl sulfate; colorants; flavorings; tablets, stabilizers; antioxidants; preservatives; pyrogen-free water; isotonic saline solution; and phosphate buffer, etc. These materials are used as needed to aid in the stability of the formulation or to help increase the activity or its bioavailability or to produce an acceptable taste or flavor in the case of oral administration.

[0088] Preferably, the drug / drug composition may also contain physiologically compatible excipients, including buffers, diluents, excipients, fillers, binders, wetting agents, disintegrants, absorption enhancers, surfactants, adsorption carriers, lubricants, etc.

[0089] Specifically, the drug / drug composition is administered orally, by injection, nasally, transdermally or through a mucosa, etc.

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

[0091] Specifically, the medicine / drug composition includes liquid dosage forms, gas dosage forms, solid dosage forms, semi-solid dosage forms, and the like.

[0092] Preferably, the administration route of the drug / drug composition is preferably parenteral administration, injection or oral administration. The injection preferably includes intravenous injection, intramuscular injection, intraperitoneal injection, intradermal injection or subcutaneous injection. The drug / drug composition is a conventional dosage form in the art, preferably in the form of solid, semi-solid, gas or liquid, that is, it can be an aqueous solution, non-aqueous solution or suspension, more preferably tablets, capsules, granules, injection or infusion, etc. More preferably, it is administered intravascularly, subcutaneously, intraperitoneally or intramuscularly. Preferably, the drug / drug composition can also be administered as an aerosol or a coarse spray, that is, nasally; or, intrathecally, intramedullary or intraventricularly. More preferably, the drug / drug composition can also be administered transdermally, percutaneously, topically, enterally, intravaginally, sublingually or rectally. The drug / drug composition of the present invention can be prepared into various dosage forms as needed, and can be administered by a physician according to factors such as the type of patient, age, weight and general disease condition, and mode of administration to determine the dosage that is beneficial to the patient. The administration method may be, for example, injection or other treatment methods.

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

[0094] The therapeutically effective dose for the drug / pharmaceutical composition of the present invention can be initially estimated in cell culture experiments or animal models such as rodents, rabbits, dogs, pigs and / or primates. Animal models can also be used to determine suitable concentration ranges and approaches for administration. Subsequently, it can be used to determine useful doses and approaches for administration in humans. Generally, the determination and adjustment of the administration of an effective amount or dosage and when and how to conduct such an adjustment assessment are known to those skilled in the art.

[0095] For additional guidance on formulation, dosage, administration schedules, 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 invention also provides compounds represented by formula (I), formula (II-1), formula (II-2), formula (III-1), formula (III-2), formula (1)-formula (26) or their stereoisomers, pharmaceutically acceptable salts, prodrugs or solvates, or the preparation methods as described above, or the use of the drugs / drug compositions as described above in the preparation of drugs for treating or preventing diseases related to the activity or expression level of DRAK2, or in the preparation of drugs for inhibiting the kinase activity of DRAK2.

[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, and biliary cirrhosis.

[0098] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features specifically described below (such as embodiments) can be combined with each other to form a new or preferred technical solution. Due to space limitations, they will not be described one by one here.

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

[0100] The present invention is further described in detail with reference to the following specific examples. The process, conditions, implementation methods, etc. of the present invention, except for the contents specifically mentioned below, are all common knowledge and common common sense in the art and are not particularly limited by the present invention.

[0101] The following description of at least one exemplary embodiment is actually only illustrative and is by no means intended to limit the present invention and its application or use. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0102] In all examples shown and discussed herein, any specific values ​​should be interpreted as merely exemplary and not as limiting. Therefore, other examples of the exemplary embodiments may have different values.

[0103] The present invention provides a novel 3-ureido-substituted thieno[2,3-b]pyridine compound, a preparation method and an application thereof. The preparation method comprises the following steps: 5-bromothieno[2,3-b]pyridine-3-amine undergoes a nucleophilic substitution reaction with phenyl chloroformate or phenyl chlorothioformate to generate an intermediate, the intermediate undergoes a nucleophilic substitution reaction with a substituted amine to generate a key intermediate, and then the key intermediate undergoes a palladium-catalyzed Suzuki coupling reaction or a Buchwald–Hartwig coupling reaction with a substituted aryl boronic acid or boronic acid ester or a substituted aryl amine under alkaline conditions to obtain a target product, 3-ureido-substituted thieno[2,3-b]pyridine compound. The 3-ureido-substituted thieno[2,3-b]pyridine compound of the present invention has certain prospects as an inhibitor of DRAK2 in the preparation of a drug for the treatment of metabolic syndrome.

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

[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) and 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 spin-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) and 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 it was complete, and appropriate amounts of water and ethyl acetate were added to the reaction system for extraction, and the organic phase was retained. After drying over anhydrous sodium sulfate, the organic phase was mixed with silica gel, and separated and purified by column chromatography (dichloromethane: methanol, v: v = 20: 1) to obtain intermediate IV-2 (66 mg). 1 H NMR (400MHz, DMSO-d6) δ8.91(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] N,N-dimethylformamide (20 mL), water (4 mL), IV-2 (150 mg, 0.425 mmol), potassium carbonate (117 mg, 0.850 mmol), p-aminophenylboronic acid (70 mg, 0.425 mmol), tetrakistriphenylphosphine palladium (49 mg, 0.0425 mmol) were added to a single-necked flask in sequence, and the system was reacted at 85°C for 12 hours, and the reaction was monitored by TLC until it was complete. An appropriate amount of water and ethyl acetate were added to the reaction system for extraction, and the organic phase was retained. After drying over anhydrous sodium sulfate, the organic phase was mixed with silica gel, and separated and purified by column chromatography (dichloromethane: methanol, v: v = 15: 1) to obtain the target compound 1 (34 mg) as a white solid. 1 H NMR(400MHz,DMSO-d6)δ8.98(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=7.8Hz,1H),3.60–3.46(m,1H),1.91–1.50(m,5H),1.40–1.17(m,5H). 13 C NMR(101MHz,DMSO-d6)δ167.89,158.62,154.82,146.10,140.41,134.11,131.0 4,129.95,128.86,127.34,126.74,126.56,106.20,48.32,33.40,25.69,24.81.

[0110] Example 2: Synthesis of Compound 2.

[0111]

[0112] The raw material p-aminoformylphenylboronic acid in Example 1 of the present invention was replaced by p-carboxyphenylboronic acid, and compound 2 was prepared using experimental steps similar to those in Example 1 of the present invention. 1 H NMR(400MHz,DMSO-d6)δ13.07(s,1H),8.96(s,1H),8.95(s,1H),8.50(s,1H),8.13(s,1H),8.10(s,1H),7.96(s ,1H),7.94(s,1H),7.71(s,1H),6.22(d,J=7.7Hz,1H),3.72–3.44(m,1H),1.98–1.46(m,5H),1.43–1.01(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] The raw material p-aminoylphenylboronic acid in Example 1 of the present invention was replaced by 5-carboxy-2-fluorophenylboronic acid, and compound 3 was prepared using experimental steps similar to those in Example 1 of the present 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] The raw material p-aminoformylphenylboronic acid in Example 1 of the present invention was replaced by p-acetylaminophenylboronic acid, and compound 4 was prepared using experimental steps similar to those in Example 1 of the present invention. 1H NMR(400MHz,DMSO-d6)δ10.12(s,1H),8.87(s,2H),8.36(s,1H),7.82–7.68(m,4H),7.67(s,1H) ),6.19(d,J=7.2Hz,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(101MHz,DMSO-d6)δ168.96,157.71,154.80,145.89,139.87,132.19,131.69,1 29.81,127.88,126.72,125.69,119.97,106.09,48.33,33.42,25.68,24.82,24.55.

[0119] Example 5: Synthesis of Compound 5.

[0120]

[0121] The raw material p-aminoformylphenylboronic acid in Example 1 of the present invention was replaced by 4-(N-methylformamido)phenylboronic acid, and compound 5 was prepared using experimental steps similar to those in Example 1 of the present invention. 1 H NMR(400MHz,DMSO-d6)δ9.13(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=7.3Hz,1H),3.60–3.48(m,1H),2.82(d,J=3.6Hz,3H),1.93–1.51(m,5H),1.42–1.13(m,5H). 13 CNMR(101MHz,DMSO-d6)δ166.60,158.58,154.92,146.01,140.17,134.26,130.95,1 30.09,128.41,127.36,126.79,126.74,105.87,48.22,33.39,26.77,25.70,24.76.

[0122] Example 6: Synthesis of compound 6.

[0123]

[0124] The raw material p-aminoformylphenylboronic acid in Example 1 of the present invention was replaced by 4-(aminosulfonyl)phenylboronic acid, and compound 6 was prepared using experimental steps similar to those in Example 1 of the present invention.1 H NMR(400MHz,DMSO-d6)δ8.97(s,1H),8.91(s,1H),8.47(s,1H),7.99(s,4H),7.71(s,1 H),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(101MHz,DMSO-d6)δ158.92,154.80,146.16,144.02,141.08,130.57,1 29.92,128.13,128.07,126.99,126.73,106.55,48.35,33.41,25.68,24.84.

[0125] Example 7: Synthesis of compound 7.

[0126]

[0127] The raw material p-aminoformylphenylboronic acid in Example 1 of the present invention was replaced by 1-isoindolone-5-boric acid pinacol ester, and compound 7 was prepared using experimental steps similar to those in Example 1 of the present invention. 1 H NMR (400MHz, DMSO-d6) δ8.95(s,1H),8.91(s,1H),8.67(s,1H),8.47(s,1H),8.00(s,1H),7.90(d,J=7.7Hz,1H),7.84(d, J=7.7Hz,1H),7.71(s,1H),6.20(d,J=7.9Hz,1H),4.48(s,2H),3.62–3.47(m,1H),1.96–1.51(m,5H),1.44–0.96(m,5H). 13 CNMR(101MHz,DMSO-d6)δ170.08,158.64,154.79,146.23,145.67,140.91,132.71,131.50,1 29.90,127.45,126.79,126.72,124.05,122.90,106.32,48.31,45.50,33.39,25.67,24.79.

[0128] Example 8: Synthesis of Compound 8.

[0129]

[0130] The raw material p-aminoylphenylboronic acid in Example 1 of the present invention was replaced by 2-methoxy-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide, and compound 8 was prepared using experimental steps similar to those in Example 1 of the present invention. 1 H NMR(400MHz, DMSO-d6)δ9.76(s,1H),9.04(s,1H),9.01(s,1H),7.96(d,J=8.0Hz,1H),7.78–7.57(m,4H),7.52 (d,J=8.0Hz,1H),6.85(d,J=7.9Hz,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(101MHz,DMSO-d6)δ166.43,158.66,158.37,155.11,145.95,141.73,132.11,130.81,1 30.43,127.32,126.90,122.38,119.23,111.15,105.10,56.83,48.01,33.34,25.77,24.61.

[0131] Example 9: Synthesis of Compound 9.

[0132]

[0133] The raw material p-aminoylphenylboronic acid in Example 1 of the present invention was replaced by 3-chloro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide, and compound 9 was prepared using experimental steps similar to those in Example 1 of the present invention. 1 HNMR(400MHz,DMSO-d6)δ9.47(s,1H),8.66(s,1H),8.57(s,1H),8.24(s,1H),8.13(s,1H),7.99(d,J=7.8H z,1H),7.80–7.55(m,3H),6.70(d,J=7.6Hz,1H),3.64–3.48(m,1H),1.91–1.47(m,5H),1.41–1.03(m,5H). 13C NMR(101MHz,DMSO-d6)δ166.56,158.62,155.02,147.31,139.71,136.08,132.45,132.3 5,130.23,129.83,129.53,129.31,127.16,126.27,105.67,48.07,33.34,25.72,24.67.

[0134] Example 10: Synthesis of compound 10.

[0135]

[0136] The raw material p-aminoylphenylboronic acid in Example 1 of the present invention was replaced with 5-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-1H-tetrazole, and compound 10 was prepared using experimental steps similar to those in Example 1 of the present invention. 1 H NMR (400MHz, DMSO-d6) δ9.04(s,1H),8.99(s,1H),8.57(s,1H),8.48(s,1H),8.12(d,J=7.2Hz,1H),8.04(d,J=7. 3Hz,1H),7.86–7.76(m,1H),7.72(s,1H),6.29(s,1H),4.04–3.93(m,1H),1.95–1.50(m,5H),1.42–1.10(m,5H). 13 C NMR(101MHz,DMSO-d6)δ158.66,154.82,146.01,138.93,130.94,130.90,130.3 5,129.96,126.96,126.78,126.61,126.02,106.29,48.29,33.39,25.67,24.81.

[0137] Example 11: Synthesis of compound 11.

[0138]

[0139] The raw material p-aminoformylphenylboronic acid in Example 1 of the present invention was replaced by 4-methylsulfonylaminophenylboronic acid, and compound 11 was prepared using experimental steps similar to those in Example 1 of the present invention. 1H NMR (400MHz, DMSO-d6) δ9.96(s,1H),8.96(s,1H),8.87(d,J=2.1Hz,1H),8.43(d,J=2.2Hz,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=7.8Hz,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(101MHz,DMSO-d6)δ157.88,155.60,154.83,145.89,138.94,133.10,131.4 3,129.88,128.55,126.75,126.00,120.54,106.07,48.29,33.41,25.68,24.81.

[0140] Example 12: Synthesis of compound 12.

[0141]

[0142] The raw material p-aminoformylphenylboronic acid in Example 1 of the present invention was replaced by 2-acetylaminopyridine-5-boric acid, and compound 12 was prepared using experimental steps similar to those in Example 1 of the present invention. 1 HNMR(400MHz,DMSO-d6)δ10.71(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=7.6Hz,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(101MHz,DMSO-d6)δ169.96,158.25,154.78,152.37,146.58,145.78,137.08,129.8 1,128.84,128.80,126.71,125.93,113.75,106.29,48.35,33.41,25.67,24.84,24.42.

[0143] Example 13: Synthesis of compound 13.

[0144]

[0145] The raw material p-aminoformylphenylboronic acid in Example 1 of the present invention was replaced by p-propionamidophenylboronic acid, and compound 13 was prepared using experimental steps similar to those in Example 1 of the present invention. 1 H NMR (400MHz, DMSO-d6) δ10.07(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= 7.6Hz,1H),3.60–3.49(m,1H),2.37(q,J=7.4Hz,2H),1.93–1.50(m,5H),1.41–1.17(m,5H),1.11(t,J=7.5Hz,3H). 13 C NMR(101MHz,DMSO-d6)δ172.66,157.63,154.93,145.76,139.90,131.98,131.56,130.00,127 .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-aminoylphenylboronic acid in Example 1 of the present invention was replaced by (1-acetylindolin-5-yl)boric acid, and compound 14 was prepared using experimental steps similar to those in Example 1 of the present invention. 1 H NMR (400MHz, DMSO-d6) δ8.98(s,1H),8.86(s,1H),8.43(s,1H),8.17(d,J=8.2Hz,1H),7.82–7.48(m,3H),6.30(d,J=7.3Hz ,1H),4.16(t,J=8.0Hz,2H),3.61–3.47(m,1H),3.24(t,J=8.0Hz,2H),2.20(s,3H),1.93–1.51(m,5H),1.41–1.13(m,5H). 13C NMR(101MHz,DMSO-d6)δ169.21,157.60,154.85,145.85,143.45,133.55,132.47,131.82,129.89, 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 raw material p-aminoformylphenylboronic acid in Example 1 of the present invention was replaced by 3-ethoxycarbonylphenylboronic acid, and compound 15 was prepared using experimental steps similar to those in Example 1 of the present invention. 1 H NMR (400MHz, DMSO-d6) δ8.93(s,2H),8.44(s,1H),8.31(s,1H),8.09(d,J=7.7Hz,1H),8.04(d,J=7.7Hz,1H),7.76– 7.63(m,2H),6.20(d,J=7.7Hz,1H),4.38(q,J=7.0Hz,2H),3.58–3.49(m,1H),1.92–1.49(m,5H),1.42–1.14(m,8H). 13 C NMR (101MHz, DMSO-d6) δ166.03,158.61,154.82,146.08,138.43,132.47,131.37,131.04,130. 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] 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), tert-butanol (20 mL) were added to a three-necked flask in sequence, and the system was heated at 100°C for 12 hours. The reaction was monitored by TLC until completion, and appropriate amounts of water and ethyl acetate were added to the reaction system for extraction. The organic phase was retained, dried over anhydrous sodium sulfate, and then the organic phase was mixed with silica gel, separated and purified by column chromatography (DCM: MeOH, v: v = 35: 1) to obtain the target compound 16 (45 mg) as a white solid. 1 H NMR(400MHz,DMSO-d6)δ8.73(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=7.7Hz,1H),3.55–3.44(m,1H),1.89–1.48(m,5H),1.38–1.10(m,5H). 13 C NMR(101MHz,DMSO-d6)δ168.51,154.94,151.12,144.06,141.17,136.77,136.23,129.8 0,129.43,127.07,119.58,119.26,115.84,115.51,106.71,48.32,33.43,25.68,24.85.

[0155] Example 17: Synthesis of Compound 17.

[0156]

[0157] The raw material p-aminobenzamide in Example 16 of the present invention was replaced by methyl 4-aminobenzoate, and compound 17 was prepared using experimental steps similar to those in Example 16 of the present invention. 1H NMR(400MHz,DMSO-d6)δ9.04(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=7.7Hz,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(101MHz,DMSO-d6)δ166.44,154.82,152.67,149.02,142.24,134.96,131.72,1 29.41,126.96,120.22,118.00,114.59,106.97,52.06,48.38,33.42,25.66,24.89.

[0158] Example 18: Synthesis of Compound 18.

[0159]

[0160] The raw material cyclohexylamine in Example 1 of the present invention was replaced by 4-aminotetrahydropyran, and compound 18 was prepared using experimental steps similar to those in Example 1 of the present invention. 1 H NMR(400MHz,DMSO-d6)δ9.43(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=7.0Hz,1H),3.92–3.70(m,3H),3.51–3.36(m,2H),1.90–1.32(m,4H). 13 CNMR(101MHz,DMSO-d6)δ167.89,158.59,155.04,145.99,140.33,134.06,13 0.92,130.09,128.85,127.24,126.90,126.85,105.97,66.19,45.83,33.58.

[0161] Example 19: Synthesis of Compound 19.

[0162]

[0163] The raw material cyclohexylamine in Example 1 of the present invention was replaced by cyclopropylamine, and compound 19 was prepared using experimental steps similar to those in Example 1 of the present invention. 1H NMR(400MHz,DMSO-d6)δ8.99(s,1H),8.72(s,1H),8.43(s,1H),7.91–7.58(m,5H ),7.46(s,1H),7.21(s,1H),6.48(s,1H),2.43–2.32(m,1H),0.50–0.08(m,4H). 13 C NMR (101MHz, DMSO-d6) δ167.87,158.58,156.51,146.03,140.35,134.06,130.94,129.95,128.85,127.27,126.93,126.89,106.72,22.91,6.86.

[0164] Example 20: Synthesis of Compound 20.

[0165]

[0166] The raw material cyclohexylamine in Example 1 of the present invention was replaced by cycloheptylamine, and compound 20 was prepared using experimental steps similar to those in Example 1 of the present invention. 1 H NMR (400MHz, DMSO-d6) δ8.97(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=4.9Hz,1H),3.80–3.67(m,1H),1.96–1.82(m,2H),1.66–1.34(m,10H). 13 C NMR(101MHz,DMSO-d6)δ167.89,158.62,154.69,146.12,140.43,134.12,131.0 6,129.92,128.87,127.36,126.70,126.47,106.16,50.53,35.28,28.18,23.96.

[0167] Example 21: Synthesis of compound 21.

[0168]

[0169] The raw material cyclohexylamine in Example 1 of the present invention was replaced by aniline, and compound 21 was prepared using experimental steps similar to those in Example 1 of the present invention. 1H NMR(400MHz, DMSO-d6)δ9.28(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(101MHz,DMSO-d6)δ167.91,158.60,152.98,146.31,140.39,139.87,134.17, 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 raw material cyclohexylamine in Example 1 of the present invention was replaced by benzylamine, and compound 22 was prepared using experimental steps similar to those in Example 1 of the present invention. 1 H NMR(400MHz,DMSO-d6)δ9.12(s,1H),8.97(s,1H),8.51(s,1H),8.14–7.85(m,5H),7.74(s ,1H),7.46(s,1H),7.35(s,4H),7.27(s,1H),6.73(t,J=4.9Hz,1H),4.38(d,J=4.9Hz,2H). 13 C NMR(101MHz,DMSO-d6)δ167.90,158.63,155.67,146.14,140.54,140.42,134.15,134 .10,131.07,129.84,128.86,127.73,127.35,126.82,126.77,126.72,107.10,43.46.

[0173] Example 23: Synthesis of compound 23.

[0174]

[0175] The raw material cyclohexylamine in Example 1 of the present invention was replaced with trans-4-aminocyclohexanol, and compound 23 was prepared using experimental steps similar to those in Example 1 of the present invention. 1H NMR(400MHz,DMSO-d6)δ8.96(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=7.6Hz,1H),4.56(d,J=4.1Hz,1H),3.57–3.39(m,2H),2.02–1.03(m,8H). 13 C NMR(101MHz,DMSO-d6)δ167.91,158.62,154.95,146.10,140.41,134.11,131.0 4,129.89,128.87,127.33,126.74,126.55,106.36,68.45,48.33,34.25,31.23.

[0176] Example 24: Synthesis of Compound 24.

[0177]

[0178] The raw material phenyl chloroformate in Example 1 of the present invention was replaced by phenyl chlorothioformate, and compound 24 was prepared using experimental steps similar to those in Example 1 of the present invention. 1 H NMR(400MHz,DMSO-d6)δ9.59(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(101MHz,DMSO-d6)δ179.90,167.90,158.52,146.26,140.37,134.14,13 1.26,129.90,128.88,128.45,127.42,115.21,52.99,32.33,25.61,25.04.

[0179] Example 25: Synthesis of Compound 25.

[0180]

[0181] The raw materials in Example 1 of the present invention were replaced by p-acetylaminophenylboronic acid and cyclohexylamine by cyclopentylamine, and compound 25 was prepared using experimental steps similar to those in Example 1 of the present invention. 1HNMR(400MHz,DMSO-d6)δ10.14(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=7.0Hz,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(101MHz,DMSO-d6)δ168.98,157.66,155.25,145.81,139.85,132.12,131.6 0,129.90,127.83,126.77,125.85,119.93,105.83,51.49,33.33,24.55,23.62.

[0182] Example 26: Synthesis of compound 26.

[0183]

[0184] The raw materials in Example 1 of the present invention were replaced by p-acetylaminophenylboronic acid and cyclohexylamine by cyclohexylmethylamine, and compound 26 was prepared using experimental steps similar to those in Example 1 of the present invention. 1 H NMR(400MHz,DMSO-d6)δ10.13(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 (101MHz, DMSO-d6) δ168.98,157.68,155.72,145.85,139.85,132.15,131.64,129. 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 the inhibitory activity of some compounds described in the present invention on DRAK2.

[0186] 1. Materials:

[0187] Some of the compounds described in the present invention (see Table 1 for details).

[0188] 2. Test principle:

[0189] DRAK2 is a serine / threonine protein kinase. ADP-Glo TM Kinase Assay (Promega; v9102) is a luminescent kinase assay kit that can detect the ADP generated in the kinase reaction. The kit can first convert the ADP generated in the reaction system into ATP, and then ATP is quantified by Ultra-Glo TM Luciferase captures and eventually converts it into light. The emitted light signal is positively correlated with the corresponding ATP. Through this method, the inhibitory activity of small molecule compounds on DRAK2 can be measured.

[0190] 3. Test method:

[0191] Before the test, some of the compounds described in the present invention were dissolved in DMSO to prepare a stock solution, which was diluted to the required concentration with culture medium when used. 1 μL of compound was added to the wells for each assay, 2 μL of kinase solution was added to each well of the assay plate except for the control wells without enzyme, 2 μL of ATP was added to each well of the assay plate, the plate was shaken and centrifuged. The assay was stopped and 2.5 μL of ADP-Glo ​​was added. TM The reagent was used to terminate the kinase reaction and consume the unconsumed ATP, leaving only ADP and a very low ATP background for incubation at room temperature for 60 minutes. For detection analysis, 5 μL of kinase detection reagent was added to convert ADP into ATP, and luciferase and luciferin were introduced to detect ATP. Luminescence was measured using Envision, and the values ​​were copied from the Envision program, and the data were processed using GraphPadPrism 5.0. The activity percentage calculation formula is Activity% = (sample Ratio-minimum value) / (maximum value-minimum value) × 100 (maximum value refers to the value of the control group with 2% DMSO; minimum value refers to the value of the control group without enzyme). By this method, the inhibitory activity data of small molecule compounds on DRAK2 can be obtained.

[0192] 4. Results and Discussion

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

[0194]

[0195]

[0196] Note: IC 50 Evaluation of the inhibitory activity of sample drugs on DRAK2, half inhibitory concentration.

[0197] The test results show that some compounds of the present invention, 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, and 25 can inhibit DRAK2 kinase activity at a level less than 100 nM. As a novel DRAK2 inhibitor, this class of compounds can be used to develop new therapeutic drugs for metabolic syndromes including but not limited to diabetes, non-alcoholic fatty liver disease, etc.

[0198] Example 28: Functional evaluation of some compounds of the present invention on glucose-stimulated insulin release (GSIS) from mouse pancreatic islets.

[0199] 1. Materials:

[0200] Some of the compounds described in the present invention (see Table 2 for details).

[0201] 2. Test principle:

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

[0203] 3. Test method:

[0204] Some of the compounds described in the present invention were dissolved in DMSO and stored at low temperature. The concentration of DMSO in the final system was controlled within the range that did not affect the detection activity. Primary mouse islets with good growth status and uniform size were inoculated into a 48-well plate at a density of 10 / well. The primary mouse islets were starved for 1 hour with KRBH buffer containing 0.1% BSA (without free fatty acids). The supernatant was discarded, and KRBH buffer containing 0.1% BSA and 2.8mM glucose and the corresponding group of compounds described in the present invention were added to stimulate under low sugar conditions for 1 hour, and the supernatant was collected for testing. Then KRBH buffer containing 0.1% BSA and 16.7mM glucose and the corresponding group of compounds described in the present invention were added to the corresponding plate wells to stimulate under high sugar conditions for 1 hour, and the supernatant was collected for testing. The insulin content in the collected supernatant was detected using the Insulin HTRF detection kit (PerkinElmer; #62INSPEC), and the signal intensity was detected by EnVisionTM. The standard curve was established through the readings of each compound well, and the concentration of each sample (ng / mL) was calculated. Then, according to the amount of islet protein in each group, the actual content of insulin in each group (ng / μg) was calculated, and the fold value was calculated.

[0205] 4. Results and Discussion

[0206] Table 2: GSIS function evaluation results of some compounds of the present invention (including but not limited to these compounds)

[0207] compound GSIS(fold) 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 multiple of insulin secretion in the experimental group relative to the blank control group at a concentration of 5 μM after high glucose treatment, and 22b is the positive control compound.

[0209] The evaluation results show that compounds 4, 5, 6, 8, 22, and 23 can effectively promote insulin secretion in mouse pancreatic islet cells, among which compounds 4, 5, 22, and 23 have significantly better insulin secretion-promoting activity than the publicly reported DRAK2 small molecule inhibitor 22b. This class of compounds exhibits excellent insulin secretion-promoting activity and can be used to develop 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 invention have the same meaning as those commonly understood by those skilled in the art of the present invention. The terms used in the specification of the present invention are only for the purpose of describing specific embodiments and are not intended to limit the present invention.

[0211] As used in the present invention, the terms "include" and "comprising" are open expressions, that is, including the contents specified in the present invention but not excluding other contents.

[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 mentioned in the present invention are cited as references in this application, just as each document is cited as reference individually. In addition, it should be understood that after reading the above teachings of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the claims attached to this application.

Claims

1. A 3-ureido-substituted thieno[2,3-b]pyridine compound or a stereoisomer, a pharmaceutically acceptable salt, a prodrug or a solvate thereof, characterized in that: The structure of the compound is shown in formula (I): in, Ring A is selected from the group consisting of phenyl, 5-9 membered heteroaryl; R1 is selected from the following group: -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 Ra is independently selected from the following group: hydrogen, cyclopropyl, C1-C6 alkyl; Rb is selected from the following group: substituted or unsubstituted C1-C6 alkyl, phenyl; R2 is selected from the following group: hydrogen, hydroxyl, amine, halogen, C1-C6 alkyl, C1-C6 alkoxy; or, the R1 and R2 located on the adjacent ring atoms of the A ring and their adjacent ring atoms together constitute: a substituted or unsubstituted 5-7 membered heterocyclyl; R3 is selected from the group consisting of cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, phenyl, pyridyl, substituted or unsubstituted benzyl, R4 is selected from the group consisting of hydrogen, methyl; L is selected from the group consisting of a covalent single bond, -NH-; X is selected from the group consisting of oxygen, sulfur; The substitution refers to the replacement of the hydrogen atoms on the group by one to three substituents selected from the group consisting of halogen, hydroxyl, amine, C1-C3 alkyl, C1-C3 alkoxy, acetyl, 2. The compound according to claim 1 or its stereoisomer, pharmaceutically acceptable salt, prodrug or solvate, characterized in that: The ring A is selected from the following group: phenyl, pyridyl; and / or, The R1 is selected from the following group: -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 said Ra is independently selected from the following group: hydrogen, methyl, ethyl, propyl, cyclopropyl, isopropyl, tert-butyl; and / or, said Rb is selected from the following group: methyl, chloromethyl, ethyl, phenyl; and / or, Said R2 is selected from the following group: hydrogen, hydroxyl, fluorine, chlorine, methyl, methoxy; and / or, The R3 is selected from the following group: cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, phenyl, pyridyl, benzyl, and / or, Said R4 is selected from the following group: hydrogen; and / or, The L is selected from the group consisting of a covalent single bond; and / or, The X is selected from the group consisting of oxygen.

3. The compound according to claim 1 or its stereoisomer, pharmaceutically acceptable salt, prodrug or solvate, characterized in that: The compound of formula (I) has a structure as shown in the following formula (II-1) or (II-2): Wherein, ring A, R1, R2, R3, R4, and X are defined as in claim 1.

4. The compound according to claim 1 or its stereoisomer, pharmaceutically acceptable salt, prodrug or solvate, characterized in that: The compound of formula (I) has a structure as shown in the following formula (II-1) or (II-2): Wherein, the definitions of R1, R2 and R3 are the same as those in claim 1.

5. The compound according to claim 1 or its stereoisomer, pharmaceutically acceptable salt, prodrug or solvate, characterized in that: The compound of formula (I) is selected from the following group:

6. A method for preparing a 3-ureido-substituted thieno[2,3-b]pyridine compound, characterized in that: The preparation method comprises the following route: (1) In the presence of a base, 5-bromothieno[2,3-b]pyridin-3-amine reacts with phenyl chloroformate or phenyl chlorothioate to produce an intermediate IV-1 by a nucleophilic substitution reaction in a first solvent; (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 an intermediate IV-2; (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 aryl boronic acid or boric acid ester under alkaline conditions to generate the target compound II-1; or, 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 reaction process of the route is shown in reaction formula (a): Wherein, R5 is B(OH)2 or B(Pin); and the definitions of ring A, R1, R2, R3, R4 and X are the same as those in claim 1.

7. The preparation method according to claim 6, characterized in that: In the step (1), the first solvent is one or more of dichloromethane, tetrahydrofuran, and dimethyl sulfoxide; and / or the ratio of the first solvent to the amount of 5-bromothieno[2,3-b]pyridine-3-amine (ml:mmol) is 1:0.02 to 1:0.04; and / or the molar ratio of 5-bromothieno[2,3-b]pyridine-3-amine to phenyl chloroformate or phenyl chlorothioformate is 1:1 to 1:1.5; and / or the base is one or more of potassium carbonate and sodium bicarbonate; 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 the step (2), the second solvent is one or more of dimethyl sulfoxide, acetonitrile, and toluene; and / or the ratio of the amount of the second solvent to the intermediate IV-1 (ml:mmol) is 1:0.02 to 1:0.04; 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 the intermediate IV-1 to the substituted amine is 1:1 to 1:1.5; and / or the base is one or more of potassium carbonate and triethylamine; and / or the molar ratio of the base to the 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 the step (3), the third solvent is one or more of dimethyl sulfoxide, N,N-dimethylformamide, toluene, and water; and / or, the ratio of the third solvent to the intermediate IV-2 (ml:mmol) is 1:0.01 to 1:0.03; and / or, the substituted aromatic boronic acid or boronic acid ester is p-aminoformylphenylboronic acid, p-carboxyphenylboronic acid, 1-isoindolone-5-boric acid pinacol ester, 5-carboxy-2-fluorophenylboric acid, p-acetamidophenylboronic acid, 4-(N-methylformylamino)phenylboronic acid, 4-(aminosulfonyl)phenylboronic 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-methylsulfonylaminobenzene one or more of boronic acid, 2-acetylaminopyridine-5-boric acid, p-propionamidophenylboric acid, (1-acetylindolin-5-yl)boric acid, and 3-ethoxycarbonylphenylboric acid; and / or, the molar ratio of the intermediate IV-2 to the substituted arylboronic acid or boric acid ester is 1:1 to 1:5; and / or, the palladium catalyst is tetrakistriphenylphosphine palladium, bistriphenylphosphine palladium dichloride (II), 1,1'-bis(diphenylphosphino)ferrocene palladium dichloride (II) dichloromethane composite one or more of the compounds; and / or, the molar ratio of the intermediate IV-2 to the palladium catalyst is 1:0.05 to 1:0.10; and / or, the base is one or more of potassium carbonate and sodium carbonate; 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 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, The fourth solvent is one or more of toluene, dioxane, and tert-butanol; in the step (3), the ratio of the amount of the fourth solvent to the intermediate IV-2 (ml:mmol) is 1:0.03 to 1:0.05; and / or, and / or, the substituted aromatic amine includes one or more of p-aminobenzamide and methyl 4-aminobenzoate; and / or, the molar ratio of the intermediate IV-2 to the substituted aromatic amine is 1:1 to 1:5; and / or, the palladium catalyst is one or more of tris(dibenzylideneacetone)dipalladium, tetrakistriphenylphosphine palladium, and bistriphenylphosphine palladium dichloride (II); and / or, the molar ratio of the intermediate IV-2 to the palladium catalyst is 1:0.05 to 1:0.10; and / or, the phosphine ligand is One or more of (S)-(-)-1,1′-binaphthyl-2,2′-bisdiphenylphosphine, 4,5-bis(diphenylphosphino)-9,9-dimethyloxanthene, and 2-dicyclohexylphosphino-2′,4′,6′-triisopropylbiphenyl; 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 base is one or more of cesium carbonate and potassium tert-butoxide; 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.

8. A drug / drug composition, characterized in that Comprising the compound described in any one of claims 1 to 5 or its stereoisomer, pharmaceutically acceptable salt, prodrug or solvate; and a pharmaceutically acceptable carrier.

9. Use of the compound according to any one of claims 1 to 5 or its stereoisomer, pharmaceutically acceptable salt, prodrug or solvate, or the preparation method according to claim 6 or 7, or the drug / drug composition according to claim 8 in the preparation of a drug for treating or preventing a disease related to DRAK2 activity or expression, or a drug for inhibiting DRAK2 kinase activity.

10. The use according to claim 9, characterized in that The disease is metabolic syndrome; wherein the metabolic syndrome includes diabetes, non-alcoholic fatty liver disease, hyperlipidemia, obesity, and biliary cirrhosis.

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