8-(Pyridylamide) Substituted Coumarin Compounds, Methods for Their Preparation and Use

Abstract

The present application discloses 8-(pyridylamide) substituted coumarin compounds, their preparation method and use. The structure of the 8-(pyridylamide) substituted coumarin compounds is as shown in formula (I), in which the A ring is independently selected from the group consisting of a phenyl group, a naphthyl group and a 5-14 membered aromatic heterocyclic group. The 8-(pyridylamide) substituted coumarin compounds according to the present application have a new compound structure and have strong SIRT2 inhibitory activity. Among them, 18 compounds have been reported to have in vitro SIRT2 inhibitory activity IC 50 reaches micromolar levels and has significant protective effect on neuroma cells.Therefore, it can be widely used in the preparation of medicaments for treating and / or preventing diseases or pathologies associated with SIRT2 overactivity or overexpression of SIRT2, or for treating and / or preventing Parkinson's disease, metabolic diseases, and tumors.

Description

[Technical field]

[0001] The present application belongs to the field of pharmaceutical technology, specifically relates to 8-(pyridylamide) substituted coumarin compounds, their preparation methods and uses, and more particularly relates to 8-(pyridylamide) substituted coumarin compounds that can inhibit the activity or expression of Sirtuin 2 (also known as SIRT2), their preparation methods and uses. [Background technology]

[0002] Parkinson's disease is a common neurodegenerative disease, and its main clinical findings are bradykinesia, rigidity and resting tremor in patients. The pathological changes of Parkinson's disease are mainly expressed in the degenerative necrosis of dopaminergic neurons in the substantia nigra of the midbrain, and the resulting significant decrease in the dopamine content in the striatum. Parkinson's disease patients bear a great physiological and psychological burden. However, at present, there are no drugs that can play a protective role against the degenerative lesions of Parkinson's disease neurons. The therapeutic drugs for Parkinson's disease on the market (e.g., L-dopa) mainly alleviate the symptoms of the disease, and taking such drugs for a long period of time will give patients serious side effects. Therefore, it is of great significance to explore new methods for treating Parkinson's disease and discover new therapeutic drugs for Parkinson's disease.

[0003] Sirtuin is a type of histone acetyltransferase, whose enzymatic activity is mainly expressed in removing acetyl groups from lysine residues in histones or other proteins. In this process, Sirtuin depends on nicotinamide adenine dinucleotide (NAD+) as one of the substrates. The gene sequences of the Sirtuin family are relatively conservative in different species. Crystal structure studies reveal that Sirtuin is composed of two domains with different sizes. The large domain has a conservative sequence and has the characteristics of NAD and NADP binding enzymes. In comparison, the small domain has a large variation in sequence. The active center of Sirtuin is located in the slit between the large and small domains. The human Sirtuin family includes seven members, SIRT1 to SIRT7, each of which is located at a different intracellular site and acts by binding to different substrates including p53, α-tubulin, FOXO, etc., thereby achieving different biological regulatory functions. The biological functions of sirtuins are a hotspot of current biomedical research, and their functions are mainly expressed in the fields related to metabolism, aging, longevity, stress response and genome stability, etc.

[0004] As an important member of the Sirtuin family, SIRT2 is mainly distributed in the cytoplasm, and its substrate proteins include α-tubulin, histone H4, p53, FOXO, and 14-3-3 protein. Functional studies of SIRT2 have shown that SIRT2 regulates the process of cell mitosis by deacetylating H4-K16, and SIRT2 mediates the regulation of the anaphase-promoting complex / cyclosome (ANAPC). It has been shown that SIRT2 maintains genome stability by activating the activity of the APC / C system, and that SIRT2 regulates the process of cell necrosis by mediating the mutual binding of Receptor-interacting protein 1 and 3 (RIP1 and RIP3). Recent studies have revealed that inhibition of SIRT2 activity has potential therapeutic value in Parkinson's disease, specifically, as shown below: (1) SIRT2 is highly expressed in the central nervous system of the adult brain and regulates the related physiological metabolism; (2) SIRT2 siRNA and small molecule inhibitor AGK2 can rescue α-synulein-induced neuronal toxicity, and AGK2 can also protect neurons in a transgenic Drosophila model of Parkinson's disease in vivo in a dose-dependent manner to reduce apoptosis; (3) SIRT2 inhibits α-synulein-induced neuronal toxicity and protects neurons in a transgenic Drosophila model of Parkinson's disease in vivo in a dose-dependent manner to reduce apoptosis; and (4) SIRT2 inhibits α-synulein-induced neuronal toxicity and protects neurons in a transgenic Drosophila model of Parkinson's disease in vivo in a dose-dependent manner to reduce apoptosis. ) SIRT2 deacetylates FOXO3a, thereby increasing the expression of Bim at the RNA and protein levels, and promoting apoptosis in a Parkinson's disease cell model treated with N-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) and cell apoptosis in the brain of a mouse model treated with MPTP. However, in the brain of a mouse in which the SIRT2 gene was knocked out, the phenomenon of significantly reducing damage to the substantia nigra-striatum was displayed. (4) In addition, the small molecule inhibitor AK-7 of SIRT2 can significantly reduce the collection of mutant Huntingtin in the brain of a transgenic mouse with another neurodegenerative disease, Huntington's disease, and can significantly improve the behavioral expression of the transgenic mouse with Huntington's disease and extend its survival time. Representative SIRT2 small molecule inhibitors in the prior art include the following: [ka]

[0005] In view of the potential activity of SIRT2 small molecule inhibitors in the treatment of Parkinson's disease, the design and synthesis of new SIRT2 inhibitors is one of the current research hotspots. Summary of the Invention [Problem to be solved by the invention]

[0006] In view of the shortcomings of the prior art, the present application aims to provide 8-(pyridylamide)-substituted coumarin compounds, their preparation methods and uses, in particular, 8-(pyridylamide)-substituted coumarin compounds that can inhibit SIRT2 activity or expression, their preparation methods and uses. The present application has developed 8-(pyridylamide)-substituted coumarin compounds with novel structures that have significant SIRT2 inhibitory activity and good selectivity for SIRT2, and can be used for the prevention and treatment of Parkinson's disease, metabolic diseases, tumors, etc. [Means for solving the problem]

[0007] For the purpose of this application, the present application adopts the following technical solutions:

[0008] In aspect 1, the present application provides a method for producing a method for manufacturing a semiconductor device comprising: 8-(pyridylamide)-substituted coumarin compounds according to formula (I): [ka] wherein each A ring is independently selected from the group consisting of a phenyl group, a naphthyl group, and a 5- to 14-membered aromatic heterocyclic group (e.g., a 5-membered aromatic heterocyclic group, a 6-membered aromatic heterocyclic group, a 7-membered aromatic heterocyclic group, an 8-membered aromatic heterocyclic group, a 9-membered aromatic heterocyclic group, a 10-membered aromatic heterocyclic group, an 11-membered aromatic heterocyclic group, a 12-membered aromatic heterocyclic group, a 13-membered aromatic heterocyclic group, and a 14-membered aromatic heterocyclic group); The phenyl group, naphthyl group, and 5- to 14-membered aromatic heterocyclic group are unsubstituted or have 1 to 5 (e.g., 1, 2, 3, 4, or 5) R a is replaced by R aeach independently represents deuterium, a halogen, a hydroxyl group, a mercapto group, an amino group, a cyano group, a nitro group, an azido group, a methylsulfonyl group, an isopropylsulfonyl group, a benzenesulfonyl group, an aminosulfonyl group, a methanesulfonate group, an isopropanesulfonate group, a benzenesulfonate group, a trifluoromethyl group, a trifluoromethyloxy group, a formylamino group, C 1~8 Alkyloxy group, C 1~8 Alkylcarbonyl group, C 1~8 Alkyloxycarbonyl group, C 1~8 Alkyl group, C 3~8 selected from the group consisting of cycloalkyl groups and phenyl groups; R a In the above, 1~8 Alkyl group, C 3~8 Cycloalkyl groups and phenyl groups are unsubstituted or have 1 to 5 (e.g., 1, 2, 3, 4, 5) R b wherein R is substituted by b each independently represents deuterium, a halogen, a hydroxyl group, a mercapto group, an amino group, a cyano group, a nitro group, an azido group, a methylsulfonyl group, an isopropylsulfonyl group, a benzenesulfonyl group, an aminosulfonyl group, a methanesulfonate group, an isopropanesulfonate group, a benzenesulfonate group, a trifluoromethyl group, a trifluoromethyloxy group, a formylamino group, and C 1~8 alkylcarbonyl groups, 8-(pyridylamide) substituted coumarin compounds are provided.

[0009] Above C 1~8 means that the number of carbon atoms in the substituent is 1, 2, 3, 4, 5, 6, 7, or 8, and C 3~8 means that the substituent has 3, 4, 5, 6, 7, or 8 carbon atoms.

[0010] The 8-(pyridylamide)-substituted coumarin compounds of the present application have a new compound structure, and their in vitro SIRT2 inhibitory activity reaches 50% or more. Among them, 18 compounds have an in vitro SIRT2 inhibitory activity IC 50 reached the micromolar level, and in particular, the IC 50 A value of 10 -7 mol / L level, and four of the compounds had IC 50 A value of 10 -8 The SIRT2 inhibitory activity reached the 5-mol / L level, and had a significant protective effect on neuroma cells. Or it can be widely used in the preparation of a medicament for treating and / or preventing a disease or pathology associated with overexpression of SIRT2, or for the preparation of a medicament for treating and / or preventing Parkinson's disease, metabolic diseases, and tumors.

[0011] Preferably, in formula (I), ring A is independently selected from the group consisting of a phenyl group, a naphthyl group, and a 5- to 14-membered aromatic heterocyclic group (e.g., a 5-membered aromatic heterocyclic group, a 6-membered aromatic heterocyclic group, a 7-membered aromatic heterocyclic group, an 8-membered aromatic heterocyclic group, a 9-membered aromatic heterocyclic group, a 10-membered aromatic heterocyclic group, an 11-membered aromatic heterocyclic group, a 12-membered aromatic heterocyclic group, a 13-membered aromatic heterocyclic group, and a 14-membered aromatic heterocyclic group); The phenyl group, naphthyl group, and 5- to 14-membered aromatic heterocyclic group are unsubstituted or have 1 to 5 (e.g., 1, 2, 3, 4, or 5) R a is replaced by R a each independently represents deuterium, a halogen, a hydroxyl group, a mercapto group, an amino group, a cyano group, a nitro group, an azido group, a methylsulfonyl group, an isopropylsulfonyl group, a benzenesulfonyl group, an aminosulfonyl group, a methanesulfonate group, an isopropanesulfonate group, a benzenesulfonate group, a trifluoromethyl group, a trifluoromethyloxy group, a formylamino group, C 1~6 Alkyloxy group, C 1~6 Alkylcarbonyl group, C 1~6 Alkyloxycarbonyl group, C 1~6Alkyl group, C 3~6 cycloalkyl groups, and phenyl groups; R a In the above, 1~6 Alkyl group, C 3~6 Cycloalkyl groups and phenyl groups are unsubstituted or have 1 to 5 (e.g., 1, 2, 3, 4, 5) R b wherein R is substituted by b each independently represents deuterium, a halogen, a hydroxyl group, a mercapto group, an amino group, a cyano group, a nitro group, an azido group, a methylsulfonyl group, an isopropylsulfonyl group, a benzenesulfonyl group, an aminosulfonyl group, a methanesulfonate group, an isopropanesulfonate group, a benzenesulfonate group, a trifluoromethyl group, a trifluoromethyloxy group, a formylamino group, and C 1~6 It is selected from the group consisting of alkylcarbonyl groups.

[0012] Above C 1~6 means that the number of carbon atoms in the substituent is 1, 2, 3, 4, 5, or 6, and C 3~6 means that the substituent has 3, 4, 5, or 6 carbon atoms.

[0013] Preferably, the structure of the 8-(pyridylamide) substituted coumarin compound is shown in formula (IA). [ka] Here, R1, R2, R3, R4, and R5 are independently hydrogen, deuterium, halogen, a hydroxyl group, a mercapto group, an amino group, a cyano group, a nitro group, an azide group, a methylsulfonyl group, or a methyl group. nyl group, isopropylsulfonyl group, benzenesulfonyl group, aminosulfonyl group, methanesulfonate group, isopropanesulfonate group, benzenesulfonate group, trifluoromethyl group, trifluoromethyloxy group, formylamino group, C 1~4 Alkyloxy group, C 1~4 Alkylcarbonyl group, C 1~4 Alkyloxycarbonyl group, C1~4 It is selected from the group consisting of an alkyl group and a phenyl group.

[0014] Above C 1~4 means that the substituent has one, two, three, or four carbon atoms.

[0015] Preferably, the structure of the 8-(pyridylamide) substituted coumarin compound is shown in formula (IB). [ka] where R1, R2, R3, R4, and R5 are independently hydrogen, deuterium, halogen, a hydroxyl group, a mercapto group, an amino group, a cyano group, a nitro group, a methylsulfonyl group, a trifluoromethyl group, a trifluoromethyloxy group, a formylamino group, C 1~4 Alkyloxy group, C 1~4 Alkylcarbonyl group, and C 1~4 alkyl groups, and X is independently selected from N or C.

[0016] Above C 1~4 means that the substituent has one, two, three, or four carbon atoms.

[0017] More preferably, the 8-(pyridylamide) substituted coumarin compound is selected from the structures shown below (each represented by a structural formula or IUPAC nomenclature).

[0018] [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4]

[0019] In an embodiment 2, the present application provides a stereoisomer of the 8-(pyridylamide) substituted coumarin compound according to the embodiment 1, a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition comprising the same.

[0020] Preferably, the pharmaceutical composition further comprises a pharma- ceutically acceptable pharmaceutical auxiliary, such as a carrier, a diluent, an excipient, a filler, an adhesive, a wetting agent, a disintegrant, an emulsifier, a dissolution aid, a solubilizer, an osmotic pressure adjuster, a surfactant, a coating material, a colorant, a pH adjuster, an antioxidant, an antibacterial agent or a buffer.

[0021] The pharma- ceutically acceptable salt of the 8-(pyridylamide) substituted coumarin compound according to the present application is a salt formed with an acid selected from hydrochloric acid, hydrobromic acid, p-toluenesulfonic acid, tartaric acid, maleic acid, lactic acid, methanesulfonic acid, sulfuric acid, phosphoric acid, citric acid, acetic acid, and trifluoroacetic acid, preferably hydrochloric acid, hydrobromic acid, p-toluenesulfonic acid, or trifluoroacetic acid.

[0022] In a third aspect, the present application provides a method for preparing an 8-(pyridylamide)-substituted coumarin compound according to the first aspect, the method comprising: [ka] and DIPEA, and then reacting with 4-(2-oxo-2H-chromene-8-carboxamido)pyridine-2-sulfonyl chloride to obtain a compound according to the present invention, wherein the range of the A ring is the same as that of embodiment 1; The reaction scheme is shown below. [ka]

[0023] Preferably, the process for preparing 4-(2-oxo-2H-chromene-8-carboxamido)pyridine-2-sulfonyl chloride comprises the steps of: (1) Mixing 8-bromocoumarin and n-butyllithium, then mixing with CO2 gas under vacuum conditions, and reacting to obtain coumarin-8-carboxylic acid; (2) mixing coumarin-8-carboxylic acid with HATU and DIPEA, then mixing with 4-aminopyridine-2-sulfonic acid and reacting to obtain 4-(2-oxo-2H-chromene-8-carboxamide)pyridine-2-sulfonic acid; (3) mixing 4-(2-oxo-2H-chromene-8-carboxamido)pyridine-2-sulfonic acid with dichlorosulfoxide and reacting to obtain 4-(2-oxo-2H-chromene-8-carboxamido)pyridine-2-sulfonyl chloride; The reaction scheme is shown below. [ka]

[0024] As a preferred technical solution of the present application, the preparation method of said 4-(2-oxo-2H-chromene-8-carboxamido)pyridine-2-sulfonyl chloride includes the following steps: [ka]

[0025] (1) n-Butyllithium (n-hexane solution) is slowly added to an anhydrous tetrahydrofuran solution of 8-bromocoumarin under nitrogen protection at -78°C. After stirring at -78°C for 3 hours, the solution is frozen in a liquid nitrogen bath and then evacuated, followed by the introduction of CO2 gas generated by the reaction of BaCO3 with concentrated sulfuric acid. The mixture is stirred at -78°C for 2 hours, and then the reaction is quenched with saturated aqueous NH4Cl solution at room temperature. The pH is adjusted to about 5 with hydrochloric acid, extracted three times with ethyl acetate, washed once with saturated saline, and the combined organic phase is dried over anhydrous sodium sulfate. Dry and concentrate. Purify on a silica gel column to obtain coumarin-8-carboxylic acid. [ka]

[0026] (2) Add coumarin-8-carboxylic acid, condensing agent HATU, DIPEA, and ultra-dry dichloromethane to a single-neck flask. Stir at room temperature. Then add 4-aminopyridine-2-sulfonic acid in dichloromethane drop by drop. After the addition is complete, stir at room temperature for 5 hours. Check the completeness of the reaction by TLC. Add water to the mixture, extract with ethyl acetate three times, wash with saturated saline, and dry with anhydrous sodium sulfate. Purify with a silica gel column to obtain 4-(2-oxo-2H-chromene-8-carboxamide)pyridine-2-sulfonic acid. [ka]

[0027] (3) First, dichlorosulfoxide is introduced into a flask and heated to 60°C. While stirring, 4-(2-oxo-2H-chromene-8-carboxamide)pyridine-2-sulfonic acid is added dropwise to the reaction solution, and the mixture is heated to 60°C. The reaction products, hydrogen chloride and sulfur dioxide, are released in gas form through a reflux condenser. After the addition is complete, the mixture is stirred at 60°C for another 2 hours until no more gas is released. Then, the excess dichlorosulfoxide is distilled off using a vacuum pump. The mixture is purified using a silica gel column to obtain 4-(2-oxo-2H-chromene-8-carboxamide)pyridine-2-sulfonyl chloride. [ka]

[0028] (4) DIPEA is added to a solution of various aromatic amines in anhydrous tetrahydrofuran. The mixture is then stirred at room temperature. 4-(2-oxo-2H-chromene-8-carboxamido)pyridine-2-sulfonyl chloride is then added to the reaction solution. The mixture is stirred at room temperature for 20 min. The reaction is detected by TLC to be complete. The product is obtained by purifying it on a silica gel column.

[0029] In an embodiment 4, the present application provides a use of the stereoisomer of the 8-(pyridylamide) substituted coumarin compound according to embodiment 1 or the 8-(pyridylamide) substituted coumarin compound according to embodiment 2, a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition comprising the same, in the preparation of a medicament for treating and / or preventing a disease or condition associated with SIRT2 overactivity or overexpression of SIRT2.

[0030] Preferably, said disease or condition associated with SIRT2 overactivity or overexpression of SIRT2 comprises Parkinson's disease, metabolic disease or tumor.

[0031] In aspect 5, the present application provides a use of a stereoisomer of the 8-(pyridylamide) substituted coumarin compound according to aspect 1 or the 8-(pyridylamide) substituted coumarin compound according to aspect 2, a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition comprising the same, in the preparation of a SIRT2 inhibitor.

[0032] In embodiment 6, the present application provides a method for treating and / or preventing a disease or condition associated with SIRT2 overactivity or overexpression, the method comprising administering to a subject in need thereof a therapeutically and / or prophylactically effective amount of an 8-(pyridylamide) substituted coumarin compound according to embodiment 1 or a stereoisomer of an 8-(pyridylamide) substituted coumarin compound according to embodiment 2, a pharma- ceutical acceptable salt thereof, or a pharmaceutical composition comprising the same.

[0033] Preferably, said disease or condition associated with SIRT2 overactivity or overexpression of SIRT2 comprises Parkinson's disease, metabolic disease or tumor.

[0034] Each aspect and feature of the present application is further described below.

[0035] Various terms and phrases used in this application have the common meanings known to those skilled in the art, but this application still desires to further explain and interpret these terms and phrases, and when the terms and phrases mentioned do not match their well-known meanings, they shall conform to the meanings described in this application. The following are definitions of various terms used in this application, and these definitions apply to the terms used throughout the specification of this application unless otherwise specified. Below, the definitions of each group of the compounds of this application are provided, and unless otherwise defined, they are used uniformly in the specification and claims.

[0036] As referred to in this application, the terms "halogen", "halogen", "halogen atom", "halogenated" and the like refer to fluorine, chlorine, bromine or iodine, and in particular fluorine, chlorine or bromine.

[0037] As referred to in this application, the term "alkyl group" refers to an alkyl group having a specified number of carbon atoms, which may be a linear or branched alkyl group, for example, the "C 1~8 "Alkyl group" means an alkyl group having 1, 2, 3, 4, 5, 6, 7, or 8 carbon atoms. 1~8 Alkyl group, C 1~7 Alkyl group, C 2~8 Alkyl group, C 2~7 Alkyl group, C 2~6 Alkyl group, C 3~8 Alkyl group, C 3~7 Alkyl group, C 3~6 The alkyl group may include a subrange of groups represented by the alkyl group and the like, and preferred specific groups are, for example, a methyl group, an ethyl group, an n-propyl group, an isopropyl group, an n-butyl group, an s-butyl group, a t-butyl group, a pentyl group, a hexyl group, a heptyl group, and an octyl group, and more preferably a methyl group and an isopropyl group. 1~6 Alkyloxy group, C 1~6 Alkylformyl group, C 1~6 Alkyloxyformyl group" or "C 1~6 "C alkyl group" 1~6"Alkyl" means an alkyl group having 1, 2, 3, 4, 5, or 6 carbon atoms; 1~5 Alkyl group, C 1~4 Alkyl group, C 2~6 Alkyl group, C 2~5 Alkyl group, C 2~4 Alkyl group, C 3~6 Alkyl group, C 3~5 Alkyl group, C 3~4 a The alkyl group may include a subrange of groups represented by the above-mentioned alkyl groups, and preferred specific groups are, for example, a methyl group, an ethyl group, an n-propyl group, an isopropyl group, an n-butyl group, an s-butyl group, a t-butyl group, a pentyl group, and a hexyl group, and more preferably a methyl group. 1~4 Alkylcarbonyl group" or "C 1~4 "C" of "alkyloxycarbonyl group" 1~4 "Alkyl" means an alkyl group having 1, 2, 3, or 4 carbon atoms; 1~4 Alkyl group, C 2~4 It may include a sub-range of groups represented by alkyl groups, etc., and preferred specific groups are, for example, methyl, ethyl, n-propyl and isopropyl groups.

[0038] As referred to herein, the term "cycloalkyl group" refers to a cyclic alkyl group having a specified number of ring carbon atoms, such as the referenced "C 3~8 "Cycloalkyl group" means a cycloalkyl group having 3, 4, 5, 6, 7, or 8 carbon atoms. 3~7 Cycloalkyl groups, C 3~4 Cycloalkyl groups, C 4~6 The cycloalkyl group may include a subrange of groups, and preferred specific groups are, for example, a cyclopropyl group, a cyclobutyl group, a cyclopentyl group, a cyclohexyl group, a cycloheptyl group, and a cyclooctyl group, and more preferably, a cyclopropyl group, a cyclopentyl group, and a cyclohexyl group. 3~6 "Cycloalkyl group" means a cycloalkyl group having 3, 4, 5, or 6 carbon atoms, 3~6 Cycloalkyl groups, C 3~5 Cycloalkyl groups, C4~5 It may include a sub-range of groups represented by cycloalkyl groups, and preferred specific groups are, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl groups, and more preferably, cyclopropyl, cyclopentyl, and cyclohexyl groups.

[0039] As referred to herein, the term "aromatic heterocyclic group" refers to a heteroaromatic system containing 1 to 4 heteroatoms, including nitrogen, oxygen and sulfur heteroatoms. As referred to herein, the term "5- to 14-membered aromatic heterocyclic group" refers to a heteroaromatic system containing 5 to 14 ring atoms. Specific examples include aryl groups containing one carbon atom and four heteroatoms selected from nitrogen, oxygen, and sulfur (preferred specific groups are, for example, a tetrazole group); aryl groups containing two carbon atoms and three heteroatoms selected from nitrogen, oxygen, and sulfur (preferred specific groups are, for example, a 1,2,3-triazole group, a 1,2,4-triazole group, an oxadiazole group, and a thiadiazole group); aryl groups containing three carbon atoms and two heteroatoms selected from nitrogen, oxygen, and sulfur (preferred specific groups are, for example, an imidazole group, a pyrazole group, an oxazole group, an isoxazole group, a thiazole group, and an isothiazole group); aryl groups containing four carbon atoms and one or two heteroatoms selected from nitrogen, oxygen, and sulfur (preferred specific groups are, for example, a pyrrolyl group, a furyl group, a thienyl group, a pyridazinyl group, a pyrimidinyl group, and a pyrazinyl group); and aryl groups containing five carbon atoms and one or two heteroatoms selected from nitrogen, oxygen, and sulfur. yellow (preferable specific groups are, for example, a pyridyl group, and preferably a pyridyl group); an aryl group containing six carbon atoms and three heteroatoms selected from nitrogen, oxygen, and sulfur (preferable specific groups are, for example, a benzotriazole group); an aryl group containing seven carbon atoms and two heteroatoms selected from nitrogen, oxygen, and sulfur (preferable specific groups are, for example, a benzimidazole group and a benzpyrazole group); an aryl group containing eight carbon atoms and one to two heteroatoms selected from nitrogen, oxygen, and sulfur (preferable specific groups are, for example, an indole group, a benzofuranyl group, a benzothienyl group, a benzopyrazinyl group, a benzopyrimidinyl group, and a benzopyridazinyl group); and an aryl group containing nine carbon atoms and one heteroatom selected from nitrogen, oxygen, and sulfur (preferable specific groups are, for example, a quinolyl group and an isoquinolyl group).

[0040] As referred to in this application, the term "effective amount" refers to an amount of a compound that is effective to treat a disease or condition of interest in a subject. It means a dose capable of achieving treatment and / or prophylaxis.

[0041] As referred to in this application, the term "pharmaceutical composition" may further mean a "composition" that can be used to achieve the treatment and / or prevention of a disease or condition according to this application in a subject, particularly a mammal.

[0042] As referred to in this application, the term "subject" may mean a patient or an animal, particularly a mammal, such as a human, dog, monkey, cow, horse, etc., that receives a compound, pharmaceutical salt, or pharmaceutical composition thereof, to treat and / or prevent a disease or condition of this application.

[0043] As referred to in the present application, the term "disease and / or condition" refers to a physical condition of the subject, which physical condition is related to the disease and / or condition of the present application. For example, the disease and / or condition of the present application may refer to a physical condition, such as a physical condition of Parkinson's disease, or may refer to a disease state, such as Parkinson's disease. In the present application, the physical condition and the disease state are not distinguished, or both can represent each other, for example, "Parkinson's disease" and "Parkinsonism" can be used interchangeably.

[0044] As referred to in the present application, the term "pharmaceutical acceptable" refers, for example, to a "pharmaceutical acceptable salt", which may refer to a synthetic substance that is physiologically acceptable to a subject, as well as a medicamentously valuable substance, for example, a salt formed as an intermediate during chiral resolution, and such an intermediate salt cannot be administered directly to a subject, but can serve to obtain the final product of the present application.

[0045] In a further aspect of the present application, the present application also relates to a pharmaceutical composition containing the compound of the present application as an active ingredient. The pharmaceutical composition can be prepared based on a method known in the art. The compound of the present application can be combined with one or more pharma- ceutically acceptable solid or liquid excipients and / or auxiliaries to prepare any dosage form suitable for use by humans or animals. The content of the compound of the present application in the pharmaceutical composition is usually 0.1 to 95% by weight.

[0046] The compounds of the present application or pharmaceutical compositions containing same can be administered in unit dose form, and the route of administration can be enteral or parenteral, for example, oral, intravenous, intramuscular, subcutaneous, nasal, oral mucosa, eye, lung and airway, skin, vagina, rectum, etc.

[0047] The dosage form may be a liquid dosage form, a solid dosage form or a semi-solid dosage form. The liquid dosage form may be a solution (including true solution and colloidal solution), an emulsion (including o / w type, w / o type and complex emulsion), a suspension, an injection (including water injection, powder injection and infusion), an eye drop, a nose drop, a lotion and a liniment, etc., the solid dosage form may be a tablet (including ordinary tablet, enteric tablet, buccal tablet, dispersible tablet, chewable tablet, effervescent tablet, oral disintegrating tablet), a capsule (including hard capsule, soft capsule, enteric capsule), a granule, a powder, a pellet, a drop, a suppository, a film, a patch, a (powder) aerosol, a spray, etc., and the semi-solid dosage form may be an ointment, a gel, a paste, etc.

[0048] The compounds of the present application may be formulated into conventional formulations, sustained release formulations, controlled release formulations, targeted formulations and microparticle delivery systems.

[0049] In order to prepare the compound of the present application into a tablet, various excipients known in the art can be widely used, including diluents, adhesives, wetting agents, disintegrants, lubricants, and dissolution aids. Diluents include starch. The adhesive may be starch slurry, dextrin, sucrose, glucose, lactose, mannitol, sorbitol, xylitol, microcrystalline cellulose, calcium sulfate, calcium hydrogen phosphate, calcium carbonate, etc.; the wetting agent may be water, ethanol, isopropyl alcohol, etc.; the adhesive may be starch slurry, dextrin, syrup, honey, glucose solution, microcrystalline cellulose, gum arabic slurry, gelatin slurry, sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, ethylcellulose, acrylic resin, carbomer, polyvinylpyrrolidone, polyethylene glycol, etc.; the disintegrant may be dry starch, microcrystalline cellulose, low-substituted hydroxypropylcellulose, crosslinked polyvinylpyrrolidone, crosslinked sodium carboxymethylcellulose, sodium carboxymethyl starch, sodium hydrogen carbonate, citric acid, polyoxyethylene sorbitan fatty acid ester, sodium dodecyl sulfonate, etc.; and the lubricant and dissolution aid may be talc powder, silica, stearate, tartaric acid, liquid paraffin, polyethylene glycol, etc.

[0050] The tablets may further be made into coated tablets, such as sugar-coated tablets, film-coated tablets, enteric-coated tablets, or double- and multi-layer tablets.

[0051] To prepare the dosage unit into a capsule, the active ingredient of the compound of the present application can be mixed with a diluent and a dissolution aid, and the mixture can be directly put into a hard or soft capsule. The active ingredient of the compound of the present application can also be made into granules or pellets with a diluent, adhesive, and disintegrant, and then put into a hard or soft capsule. The varieties of diluents, adhesives, wetting agents, disintegrants, and dissolution aids used to prepare the compound tablets of the present application can also be used to prepare the capsules of the compound of the present application.

[0052] In order to prepare the compound of the present application into an injection, water, ethanol, isopropyl alcohol, propylene glycol or a mixture thereof can be used as a solvent, and a suitable amount of a solubilizer, dissolution aid, pH adjuster, or osmotic pressure adjuster commonly used in the art can be added. The solubilizer or dissolution aid can be poloxamer, lecithin, hydroxypropyl-β-cyclodextrin, etc., the pH adjuster can be phosphate, acetate, hydrochloric acid, sodium hydroxide, etc., and the osmotic pressure adjuster can be sodium chloride, mannitol, glucose, phosphate, acetate, etc. When preparing a lyophilized powder injection, mannitol, glucose, etc. can also be added as a proppant.

[0053] Furthermore, if necessary, coloring agents, preservatives, fragrances, flavoring agents or other additives may be added to the pharmaceutical preparations.

[0054] To achieve the purpose of drug administration and enhance the therapeutic effect, the drug or pharmaceutical composition of the present application can be administered by any known administration method.

[0055] The dosage of the pharmaceutical composition of the compound of the present application can vary widely depending on the nature and severity of the disease to be prevented or treated, the individual condition of the patient or animal, the administration route and dosage form, etc. In general, the compound of the present application has a suitable daily dosage range of 0.001-150 mg / kg body weight, preferably 0.1-100 mg / kg body weight, more preferably 1-60 mg / kg body weight, and most preferably 2-30 mg / kg body weight. The above dosage can be administered in one dosage unit or divided into several dosage units, and is determined by the doctor's clinical experience and the administration mode including the use of other treatment means.

[0056] The compounds or compositions of the present application may be administered alone or in combination with other therapeutic or symptomatic drugs. When the compounds of the present application have synergistic effects with other therapeutic drugs, the synergistic effects may be adjusted according to the actual situation. The dose should be adjusted accordingly. Effect of the Invention

[0057] Compared with the prior art, the present application has the following beneficial effects:

[0058] The 8-(pyridylamide)-substituted coumarin compounds of the present application have a new compound structure, and their in vitro SIRT2 inhibitory activity reaches 50% or more. Among them, 18 compounds have an in vitro SIRT2 inhibitory activity IC 50 reached the micromolar level, and in particular, the IC 50 A value of 10 -7 mol / L level, and four of the compounds had IC 50 A value of 10 -8 The SIRT2 inhibitory activity reaches 1000 mol / L level, exhibits good SIRT2 inhibitory activity, and has a significant protective effect on neuroma cells. Therefore, it can be widely used in the preparation of medicaments for treating and / or preventing diseases or pathologies associated with SIRT2 overactivity or SIRT2 overexpression, or in the preparation of medicaments for treating and / or preventing Parkinson's disease, metabolic diseases, and tumors. [Brief description of the drawings]

[0059] [Figure 1] FIG. 2 is a statistical diagram showing the results of protective action of the 8-(pyridylamide)-substituted coumarin compounds of the present application against SH-SY5Y cell damage in Test Example 3. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0060] The technical solution of the present application will be further described below through specific embodiments. Those skilled in the art should understand that the above examples are only for understanding the present application, and should not be regarded as specific limitations of the present application.

[0061] For all of the following examples or preparations, standard procedures for operation and purification well known to those skilled in the art can be used. Unless otherwise specified, all temperatures are expressed in degrees Celsius, and the structures of compounds were confirmed by nuclear magnetic resonance spectroscopy (NMR) and / or mass spectrometry (MS).

[0062] In all of the following examples or preparations, the structures of the compounds are shown by nuclear magnetic resonance hydrogen spectrum ( 1 The NMR spectra were determined by H NMR (H NMR) or mass spectrometry (MS). Nuclear magnetic resonance hydrogen spectral shifts (δ) are given in parts per million (ppm). Nuclear magnetic resonance spectra were recorded on a Mercury-300 or Mercury-400 nuclear magnetic resonance spectrometer in deuterated chloroform (CDCl3) or deuterated dimethyl sulfoxide (DMSO-d6) with tetramethylsilane (TMS) or sodium 3-(trimethylsilyl) deuterated propionate (TSM) as the internal standard.

[0063] The electronic balance used was a Japanese Yanaco LY-300 type electronic balance.

[0064] In the column chromatography, 200-300 mesh or 300-400 mesh silica gel was used as a carrier.

[0065] All anhydrous solvents were handled by standard methods. All other reagents were of commercial analytical purity.

[0066] Where: HATU was 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluor-ophosphat, i.e., 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate.

[0067] DIPEA is N,N-diisomromylethylamine, i.e., N,N-diisopropylethylamine.

[0068] DMF was N,N-dimethylformamide.

[0069] The origins of the raw materials used in the following Preparation Examples and Examples are as follows: 8-Bromocoumarin was purchased from Shanghai Aladdin Biochemical Technology Co., Ltd., CAS: 33491-30-4; 4-Aminopyridine-2-sulfonic acid was purchased from SYNCHEM-OHG, CAS: 900804-14-0; 4-Chloroaniline, 3-chloroaniline, 2-chloroaniline, 4-methylaniline, 3-nitroaniline, 2-cyanoaniline, 4-fluoroaniline, 4-methylsulfonylaniline, 4-trifluoromethylaniline, 4-phenylaniline, 4-hydroxylaniline, 4-methoxyaniline, 3,4-fluoroaniline, 3-bromo-4-chloroaniline, 7-bromo-1-naphthylamine, 3-chloro-1-naphthylamine, 5-aminoquinoline, and 3-bromo-5-aminoquinoline were obtained from Macklin (Shanghai) Biochemical Technology Co., Ltd., Sigma-Aldrich (Shanghai) Trading Co., Ltd., J&K Scientific (Beijing) Co., Ltd., Meryer (Shanghai) Chemical Technology Co., Ltd., and Alfa They were purchased from Aesar (China) Chemical Co., Ltd., Beijing Gouhe Technology Co., Ltd., Shanghai Aladdin Biochemical Technology Co., Ltd., SolAlbio Science & Technology (Beijing) Co., Ltd., and InnoChem (Beijing) Technology Co., Ltd.

[0070] [Preparation Example 1] In this preparation example, intermediate 4 represented by the following formula was prepared, and the synthetic route was as follows. [ka]

[0071] (1) n-Butyllithium (1.60M n-hexane solution) (14.00mL, 22.33mmol) was slowly added to a solution of 8-bromocoumarin (5.00g, 22.33mmol) in anhydrous tetrahydrofuran (30mL) under nitrogen protection at -78℃. After stirring at -78℃ for 3 hours, the solution was frozen in a liquid nitrogen bath and then evacuated, followed by the introduction of CO2 gas generated by the reaction of BaCO3 (5.29g, 26.80mmol) with concentrated sulfuric acid (1.43mL, 26.80mmol). The mixture was stirred at -78℃ for 2 hours, and then the reaction was quenched with saturated aqueous NH4Cl solution at 20℃. The pH was adjusted to 5 with hydrochloric acid, extracted three times with ethyl acetate (15mL×3), washed once with saturated brine, and the combined organic phase was dried over anhydrous sodium sulfate and concentrated. The product, coumarin-8-carboxylic acid (1.61 g, 38% yield), was obtained by purification on a silica gel column.

[0072] (2) Coumarin-8-carboxylic acid (1.60 g, 8.42 mmol), condensation agent HATU (3.20 g, 8.42 mmol), DIPEA (2.93 mL, 16.84 mmol), and ultra-dry dichloromethane (15 mL) were added to a one-neck flask. Stir at 20°C for 30 min. Then, a solution of 4-aminopyridine-2-sulfonic acid (1.33 g, 7.65 mmol) in dichloromethane (5 mL) was added dropwise. After the addition was completed, the mixture was stirred at 20° C. for 5 hours. The completeness of the reaction was detected by TLC. Water was added to the mixture, which was then extracted three times with ethyl acetate (15 mL×3), washed with saturated saline, and dried over anhydrous sodium sulfate. The product, 4-(2-oxo-2H-chromene-8-carboxamido)pyridine-2-sulfonic acid (1.61 g, 61% yield), was obtained by purifying it with a silica gel column.

[0073] (3) First, dichlorosulfoxide (5 mL) was introduced into the flask and heated to 60°C. While stirring, 4-(2-oxo-2H-chromene-8-carboxamide)pyridine-2-sulfonic acid (1.60 g, 4.62 mmol) was added dropwise to the reaction solution within 3 hours, and the mixture was heated to 60°C. The reaction products, hydrogen chloride and sulfur dioxide, were released in gas form through the reflux condenser. After the dropwise addition was completed, the mixture was stirred again at 60°C for 2 hours until no gas was released. Then, the excess dichlorosulfoxide was distilled off with a vacuum pump. The product, 4-(2-oxo-2H-chromene-8-carboxamide)pyridine-2-sulfonyl chloride (0.59 g, 35% yield), was obtained by purifying it with a silica gel column.

[0074] [Example 1] In this example, compound 1, N-(2-(N-(4-chlorophenyl)sulfonamido)-pyridin-4-yl)-2-oxo-2H-chromene-8-amide, was prepared, and its structure is as follows: [ka]

[0075] DIPEA (0.48mL, 2.74mmol) was added to a solution of 4-chloroaniline (0.17g, 1.37mmol, CAS:106-47-8) in anhydrous tetrahydrofuran (10mL). The mixture was then stirred at 20°C for 10min. 4-(2-oxo-2H-chromene-8-carboxamido)pyridine-2-sulfonyl chloride (0.50g, 1.37mmol) was then added to the reaction mixture. The mixture was stirred at 20°C for 20min. The reaction was detected complete by TLC. The product was obtained by purification on a silica gel column (0.21g, 33% yield). The product was characterized as follows:

[0076] 1 H NMR (400 MHz, CDCl3): δ 8.54 (d, J = 4.4 Hz, 1H), 7.52-7.79 (m, 4H), 7.10-7.41 (m, 5H), 6.52 (m, 1H), 5.97 (m, 1H). HR-MS (ESI): [M+H] + C21H15ClN3O5S Calculated value 456.0421, found value 456.0454.

[0077] [Example 2] In this example, compound 2, N-(2-(N-(3-chlorophenyl)sulfonamido)-pyridin-4-yl)-2-oxo-2H-chromene-8-amide, was prepared, and its structure is as follows: [ka]

[0078] The preparation was carried out with reference to the preparation method in Example 1, except that 4-chloroaniline was replaced with 3-chloroaniline to obtain 0.16g of white solid, with a yield of 26%. The product was characterized as follows:

[0079] 1 H NMR (400 MHz, CDCl3): 8.53 (d, J = 5.4 Hz, 1H), 7.52-7.79 (m, 4H), 7.10-7.38 (m, 5H), 6.52 (m, 1H), 5.97 (m, 1H). HR-MS (ESI): [M+H] + C 21 H 15 ClN3O5S Calculated value 456.0421, found value 456.0434.

[0080] [Example 3] In this example, compound 3, N-(2-(N-(2-chlorophenyl)sulfonamido)-pyridin-4-yl)-2-oxo-2H-chromene-8-amide, was prepared, and its structure is as follows: [ka]

[0081] The preparation was carried out with reference to the preparation method in Example 1, except that 4-chloroaniline was replaced with 2-chloroaniline to obtain 0.26g of white solid, with a yield of 42%. The product was characterized as follows:

[0082] 1 H NMR (400 MHz, CDCl3): δ 8.54 (d, J = 4.4 Hz, 1H), 7.52-7.79 (m, 4H), 7.15-7.38 (m, 5H), 6.52 (m, 1H), 5.97 (m, 1H). HR-MS (ESI): [M+H] + C 21 H 15 ClN3O5S Calculated value 456.0421, found value 456.0414.

[0083] [Example 4] In this example, compound 4, N-(2-(N-(4-methylphenyl)sulfonamido)-pyridin-4-yl)-2-oxo-2H-chromene-8-amide, was prepared, and its structure is as follows: [ka]

[0084] The preparation was carried out with reference to the preparation method in Example 1, except that 4-chloroaniline was replaced with 4-methylaniline to obtain 1.97g of white solid, with a yield of 33%. The product was characterized as follows:

[0085] 1 H NMR (400 MHz, CDCl3): δ 8.54 (d, J = 4.4 Hz, 1H), 7.53-7.78 (m, 4H), 7.12-7.49 (m, 5H), 6.52 (m, 1H), 5.97 (m, 1H), 2.34 (s, 3H). HR-MS (ESI): [M+H]+ C 22 H 18 N3O5S calculated value 436.0967, measured value 436.0945.

[0086] [Example 5] In this example, compound 5, N-(2-(N-(3-nitrophenyl)sulfonamido)-pyridin-4-yl)-2-oxo-2H-chromene-8-amide, was prepared, and its structure is as follows: [ka]

[0087] The preparation was carried out with reference to the preparation method in Example 1, except that 4-chloroaniline was replaced with 3-nitroaniline to obtain 0.23g of white solid, with a yield of 36%. The product was characterized as follows:

[0088] 1 H NMR (400 MHz, CDCl3): δ 8.53 (d, J = 5.4 Hz, 1H), 7.50-7.77 (m, 4H), 7.10-7.36 (m, 5H), 6.54 (m, 1H), 5.92 (m, 1H). HR-MS (ESI): [M+H] + C 21 H 15 N4O7S calculated value 467.0661, measured value 467.0646.

[0089] [Example 6] In this example, compound 6, N-(2-(N-(2-cyanophenyl)sulfonamido)-pyridin-4-yl)-2-oxo-2H-chromene-8-amide, was prepared and its structure The structure is as follows: [ka]

[0090] The preparation was carried out with reference to the preparation method in Example 1, except that 4-chloroaniline was replaced with 2-cyanoaniline to obtain 0.153g of white solid, with a yield of 25%. The product was characterized as follows:

[0091] 1 H NMR (400 MHz, CDCl3): δ8.53 (d, J = 5.2 Hz, 1H), 7.52-7.75 (m, 4H), 7.10-7.42 (m, 5H), 6.52 (m, 1H), 5.97 (m, 1H). HR-MS (ESI): [M+H] + C 22 H 15 N4O5S calculated value 447.0763, measured value 447.0742.

[0092] [Example 7] In this example, compound 7, N-(2-(N-(4-fluorophenyl)sulfonamido)-pyridin-4-yl)-2-oxo-2H-chromene-8-amide, was prepared, and its structure is as follows: [ka]

[0093] The preparation was carried out with reference to the preparation method in Example 1, except that 4-chloroaniline was replaced with 4-fluoroaniline to obtain 0.17g of white solid, with a yield of 29%. The product was characterized as follows:

[0094] 1 H NMR (400 MHz, CDCl3): δ 8.62 (d, J = 4.7 Hz, 1H), 7.52-7.79 (m, 4H), 7.15-7.42 (m, 5H), 6.52 (m, 1H), 5.97 (m, 1H). HR-MS (ESI): [M+H] + C 21 H 15 FN3O5S Calculated value 440.0716, Measured value 440.0740.

[0095] [Example 8] In this example, compound 8, N-(2-(N-(4-methylsulfonylphenyl)sulfonamido)-pyridin-4-yl)-2-oxo-2H-chromene-8-amide, was prepared. and its structure is as follows: [ka]

[0096] The preparation was carried out with reference to the preparation method in Example 1, and 4-chloroaniline was replaced with 4-methylsulfonylaniline to obtain 0.27g of white solid, with a yield of 39%. The product was characterized as follows:

[0097] 1 H NMR (400 MHz, CDCl3): δ 8.62 (d, J = 4.7 Hz, 1H), 7.52-7.77 (m, 4H), 7.11-7.38 (m, 5H), 6.51 (m, 1H), 5.93 (m, 1H), 3.32 (s, 3H). HR-MS (ESI): [M+H]+ C 22 H 18 N3O7S2 calculated value 500.0586, measured value 500.0594.

[0098] [Example 9] In this example, compound 9, N-(2-(N-(4-trifluoromethylphenyl)sulfonamido)-pyridin-4-yl)-2-oxo-2H-chromene-8-amide, was prepared, and its structure is as follows: [ka]

[0099] The preparation was carried out with reference to the preparation method in Example 1, and 4-chloroaniline was replaced with 4-trifluoromethylaniline to obtain 0.26g of white solid, with a yield of 39%. The product was characterized as follows:

[0100] 1 H NMR (400 MHz, CDCl3): δ 8.62 (d, J = 4.7 Hz, 1H), 7.58-7.79 (m, 4H), 7.11-7.49 (m, 5H), 6.52 (m, 1H), 5.76 (m, 1H). HR-MS (ESI): [M+H]+ C 22 H 15 F3N3O5S calculated value 490.0685, measured value 490.0683.

[0101] [Example 10] In this example, compound 10, N-(2-(N-(4-phenylphenyl)sulfonamide), (mido)-pyridin-4-yl)-2-oxo-2H-chromene-8-amide was prepared, the structure of which is as follows: [ka]

[0102] The preparation was carried out with reference to the preparation method in Example 1, except that 4-chloroaniline was replaced with 4-phenylaniline to obtain 0.28g of white solid, with a yield of 41%. The product was characterized as follows:

[0103] 1 H NMR (400 MHz, CDCl3): δ 8.62 (d, J = 4.7 Hz, 1H), 8.09-8.23 (m, 2H), 7.55-7.89 (m, 10H), 6.52-6.78 (m, 2H), 5.76-6.12 (m, 2H). HR-MS (ESI): [M+H]+ C 27 H 20 N3O5S calculated value 498.1124, measured value 498.1145.

[0104] [Example 11] In this example, compound 11, N-(2-(N-(4-hydroxylphenyl)sulfonamido)-pyridin-4-yl)-2-oxo-2H-chromene-8-amide, was prepared, and its structure is as follows: [ka]

[0105] The preparation was carried out with reference to the preparation method in Example 1, substituting 4-hydroxylaniline for 4-chloroaniline to obtain 0.12g of white solid, with a yield of 21%. The product was characterized as follows:

[0106] 1 H NMR (400 MHz, CDCl3): δ 8.72 (d, J = 4.9 Hz, 1H), 7.62-7.77 (m, 4H), 7.12-7.56 (m, 5H), 6.42 (m, 1H), 5.66 (m, 1H), 5.45 (s, 1H). HR-MS (ESI): [M+H]+ C 21 H 16 N3O6S Calculated value 438. 0760, actual value 438.0788.

[0107] [Example 12] In this example, compound 12, N-(2-(N-(4-methoxyphenyl)sulfonamido)-pyridin-4-yl)-2-oxo-2H-chromene-8-amide, was prepared, and its structure is as follows: [ka]

[0108] The preparation was carried out with reference to the preparation method in Example 1, except that 4-chloroaniline was replaced with 4-methoxyaniline to obtain 0.16g of white solid, with a yield of 26%. The product was characterized as follows:

[0109] 1H NMR (400 MHz, CDCl3): δ 8.72 (d, J = 4.5 Hz, 1H), 7.62-7.76 (m, 4H), 7.12-7.66 (m, 5H), 6.42 (m, 1H), 5.66 (m, 1H), 3.83 (s, 3H). HR-MS (ESI): [M+H]+ C 22 H 18 N3O6S calculated value 452.0916, measured value 452.0933.

[0110] [Example 13] In this example, compound 13, N-(2-(N-(3,4-difluorophenyl)sulfonamido)-pyridin-4-yl)-2-oxo-2H-chromene-8-amide, was prepared, and its structure is as follows: [ka]

[0111] The preparation was carried out with reference to the preparation method in Example 1, except that 4-chloroaniline was replaced with 3,4-fluoroaniline to obtain 0.22g of white solid, with a yield of 36%. The product was characterized as follows:

[0112] 1 H NMR (400 MHz, CDCl3): δ 8.62 (d, J = 5.7 Hz, 1H), 7.57-7.77 (m, 4H), 7.13-7.49 (m, 4H), 6.62 (m, 1H), 5.93 (m, 1H). HR-MS (ESI): [M+H]+ C 21 H 14 F2N3O5S calculated value 458.0622, measured value 458.0636.

[0113] [Example 14] In this example, compound 14, N-(2-(N-(3-bromo-4-chlorophenyl)sulfonamido)-pyridin-4-yl)-2-oxo-2H-chromene-8-amide, was prepared, and its structure is as follows: [ka]

[0114] The preparation was performed with reference to the preparation method in Example 1, except that 4-chloroaniline was replaced with 3-bromo-4-chloroaniline to obtain 0.23g of white solid, with a yield of 32%. The product was characterized as follows:

[0115] 1 H NMR (400 MHz, CDCl3): δ 8.62 (d, J = 5.7 Hz, 1H), 7.53-7.78 (m, 4H), 7.15-7.41 (m, 4H), 6.62 (m, 1H), 5.93 (m, 1H). HR-MS (ESI): [M+H]+ C 21 H 14 BrClN3O5S calculated value 533.9526, found value 533.9554.

[0116] [Example 15] In this example, compound 15, N-(2-(N-(7-bromonaphthalen-1-yl)sulfonamido)-pyridin-4-yl)-2-oxo-2H-chromene-8-amide, was prepared, and its structure is as follows: [ka]

[0117] The preparation was carried out with reference to the preparation method in Example 1, and 4-chloroaniline was replaced with 7-bromo-1-naphthylamine to obtain 0.19g of white solid, with a yield of 26%. The product was characterized as follows:

[0118] 1H NMR (400 MHz, CDCl3): δ 8.56 (d, J = 54 Hz, 1H), 8.10-8.13 (m, 3H), 7.52-7.81 (m, 4H), 7.02-7.45 (m, 4H), 6.67-6.89 (m, 2H). HR-MS (ESI): [M+H]+ C 25 H 17 BrN3O5S calculated value 550.0072, found value 550.0098.

[0119] [Example 16] In this example, compound 16, N-(2-(N-(3-chloronaphthalen-1-yl)sulfonamido)-pyridin-4-yl)-2-oxo-2H-chromene-8-amide, was prepared, and its structure is as follows: [ka]

[0120] The preparation was carried out with reference to the preparation method in Example 1, except that 4-chloroaniline was replaced with 3-chloro-1-naphthylamine to obtain 0.20g of white solid, with a yield of 29%. The product was characterized as follows:

[0121] 1 H NMR (400 MHz, CDCl3): δ 8.56 (d, J = 54 Hz, 1H), 8.11-8.15 (m, 3H), 7.55-7.87 (m, 4H), 7.02-7.45 (m, 4H), 6.69-6.92 (m, 2H). HR-MS (ESI): [M+H]+ C 25 H 17 ClN3O5S Calculated value 506.0577, found value 506.0599.

[0122] [Example 17] In this example, compound 17, N-(2-(N-(quinolin-5-yl)sulfonamido)-pyridin-4-yl)-2-oxo-2H-chromene-8-amide, was prepared, and its structure is as follows: [ka]

[0123] The preparation was carried out with reference to the preparation method in Example 1, except that 4-chloroaniline was replaced with 5-aminoquinoline to obtain 0.22g of white solid, with a yield of 34%. The product was characterized as follows:

[0124] 1 H NMR (400 MHz, CDCl3): δ 8.35-8.58 (m, 5H), 8.19 (m, 1H), 7.51-7.87 (m, 5H), 7.31 (m, 1H), 6.52 (m, 1H), 5.57 (m, 1H). HR-MS (ESI): [M+H]+ C 24 H 17 N4O5S calculated value 473.0920, measured value 473.0934.

[0125] [Example 18] In this example, compound 18, N-(2-(N-(7-bromo-quinolin-5-yl)sulfonamido)-pyridin-4-yl)-2-oxo-2H-chromene-8-amide, was prepared, and its structure is as follows: [ka]

[0126] The preparation was carried out with reference to the preparation method in Example 1, substituting 3-bromo-5-aminoquinoline for 4-chloroaniline to obtain 0.24g of white solid, with a yield of 32%. The product was characterized as follows:

[0127] 1H NMR (400 MHz, CDCl3): δ 8.31-8.52 (m, 4H), 8.13 (m, 1H), 7.51-7.82 (m, 5H), 7.31 (m, 1H), 6.52 (m, 1H), 5.57 (m, 1H). HR-MS (ESI): [M+H]+ C 24 H 16 BrN4O5S calculated value 551.0025, found value 551.0043.

[0128] [Test Example 1] Screening of SIRT2 inhibitors for in vitro activity: The full length expression sequence ORF of human SIRT2 gene is 1170 bp (Accession No.: NM_012237), and the size of the expressed SIRT2 protein is 43KDa. The expression and purification steps of SIRT2 recombinant protein are as follows: SIRT2 gene was cloned into expression carrier pET-15b (primers are as follows: Forward: 5'-TAATACGACTCACTATAGGG-3' (SEQ ID NO. 1), backward: 5'-TTCACTTCTGAGTTCGGCATG-3' (SEQ ID NO. 2)), and the expressed SIRT2 protein contains a tag His6 at the N-terminus for purification. The plasmid was transformed into E. coli BL2 (DE3), induced by 1 mM IPTG, and expressed at 18°C ​​for 6 hours. The induced E. coli cells were collected by centrifugation and frozen at -20°C. The bacteria were precipitated and suspended in 15 mL of cell lysis solution (50 mM Tris-HCl, pH 8.0, 300 mM NaCl), and the bacterial sample was sonicated for 10 min, followed by centrifugation (12000 × g, 20 min) at 4 °C to remove the precipitate. The recombinantly expressed soluble SIRT2 protein was present in the supernatant. The supernatant was then immersed in Ni 2+ The purification was performed using an NTA-agarose matrix column (Qiagen). Unbound proteins were washed with 50 mM Tris-HCl, pH 8.0, 300 mM NaCl, then non-specifically bound proteins were washed with gradually increasing concentrations of imidazole (0-200 mM), and finally purified recombinant SIRT2 protein was obtained. Imidazole was removed using a PD-10 column, and the purified SIRT2 protein concentration was measured by the Bradford method. The recombinant SIRT2 protein was stored in a storage solution (50 mM Tris-HCl, pH 8.0, 265 mM NaCl, 0.2 mM DTT, and 10% glycerol) and placed at -20°C.

[0129] According to literature reports, a method for measuring SIRT2 enzyme activity was established. First, the active substrate peptide segment of SIRT2, Ac-Gln-Pro-Lys-[Lys-(Ac)]-AMC, was synthesized and used as the substrate in the enzyme activity measurement method. The linked fluorescent tag was AMC (7-Amino-4-methylcoumarin). The whole measurement process included two steps. The catalytic reaction was carried out in 60 μL of reaction solution (25 mM Tris-HCl, pH 8.0, 137 mM NaCl, 2.7 mM KCl, 1 mM MgCl2 and 1 mg / ml BSA), and 500 μM NAD + , 50 μM of the substrate peptide segment, 1.0 μg of SIRT2, and different concentrations of compounds 1 to 18 were added. The reaction was allowed to stand at 37 °C for 2 hours. In this reaction, the acetyl groups of the lysine residues in the low molecular weight peptide segments were removed to different degrees. Then, 60 μL of sample treatment solution (50 mM Tris-HCl, pH 8.0, 100 mM NaCl, trypsin, and 4 mM nicotinamide) was added to the reaction solution, mixed, and allowed to stand at 37 °C for 20 min. The plate reader was set to excitation light 355 nm and absorption light 460 nm, and the absorption intensity was measured. For each compound under test, there were two parallel tests for each concentration, and reasonable controls were set, and the experimental results were processed using the processing software GraphPad Prism to calculate the inhibitory activity IC of each inhibitor. 50 The results are shown in Table 1 (where AGK was the positive control group).

[0130] [Table 2]

[0131] As can be seen from the data in Table 1, the 18 tested compounds had in vitro SIRT2 inhibitory activity IC 50 All of the compounds reached the micromolar level, and the IC 50 The value is 10 -7 mol / L level, and four of the compounds had IC 50 The value is 10 -8 The results showed that the 8-(pyridylamide)-substituted coumarin compounds of the present application had good SIRT2 inhibitory activity.

[0132] [Test Example 2] Measurement of cytotoxicity of compounds against SH-SY5Y neuroma cells: In this test example, the cytotoxicity of the compound against SH-SY5Y neuroma cells is measured, and the dose is determined for the next Parkinson's cell model protection test. The specific experimental steps for measuring the cytotoxicity are as follows: Cultured SH-SY5Y neuroma cells were seeded in a 96-well cell culture plate at about 6,000 cells per well. After overnight culture, the culture medium was replaced with a new one, and different concentrations of compounds 1 to 18 (0.01, 0.05, 0.1, 0.5, 1, 5, 10, 20, 50, 100, 200 μM) was added to each well, and different control samples were set up. The cells were incubated for 48 hours at 37°C. 10 μL of the dye AlamarBlue® (Invitrogen) was added to each well, and incubation was continued for about 2 hours at 37°C, and the color change of the dye solution could be observed, and the change in absorbance value was read with a plate reader (Ex: 530 nm, Em: 590 nm). The compounds under test had two parallel tests for each concentration. The experimental results were analyzed by the data processing software GraphPad Prism to calculate the cytotoxic CC of each compound. 50 The results are shown in Table 2 (where Taxol was the positive control).

[0133] [Table 3]

[0134] As can be seen from the data in Table 2, the cytotoxicity IC50 of all 18 tested compounds against SH-SY5Y was greater than 20 μM, i.e., the 8-(pyridylamide)-substituted coumarin compounds of the present application have no obvious inhibitory activity against SH-SY5Y cells.

[0135] [Test Example 3] Screening for compounds with protective activity against Parkinson's disease cell models: For screening tests of protective effects in Parkinson's disease cell models, the currently accepted neurotoxic agent MPP + A Parkinson's disease cell model was constructed by acting on neuroma cells SH-SY5Y using MPP (5 mM). This type of neurotoxic agent acts on SH-SY5Y cells, affecting the abnormal accumulation of α-synuclein in the cells and causing neurotoxicity similar to Parkinson's disease. The specific experimental method is as follows. Approximately 20,000 SH-SY5Y cells were seeded in a 96-well cell plate and cultured overnight. The control test was performed without adding any neurotoxic agent, and the other groups were treated with the neurotoxic agent MPP + (5 mM) was added to the MPP+ group, and no protective compound was added to the MPP+ group. Compounds 1 to 18 (10 μM) (AGK was the positive control) were added to the culture wells of the protection test at the same time, and the culture was continued for 48 hours. The protective effect on the Parkinson's disease cell model was evaluated by measuring the cell activity of the Parkinson's disease cell model. Data analysis of the test results was performed using GraphPad Prism. The results are shown in Figure 1.

[0136] As can be seen from Figure 1, all 18 tested compounds exhibited MPP activity to some degree. +The 8-(pyridylamide) substituted coumarin compounds of the present application can protect SH-SY5Y cells from damage caused by neurotoxic agents, and the protective effects of compounds 1, 13, 14, and 15 on neuroma cells are more pronounced.

[0137] The present application describes the 8-(pyridylamide) substituted coumarin compounds of the present application, their preparation methods and uses through the above examples, but the present application is not limited to the above examples, that is, the present application does not mean that it must be carried out depending on the above examples. Those skilled in the art should understand that any improvements to the present application, equivalent replacement of each raw material of the product of the present application, addition of auxiliary ingredients, selection of specific forms, etc. are all within the protection scope and disclosure scope of the present application.

[0138] Although the preferred embodiments of the present application have been described in detail above, the present application is not limited to the specific details in the above embodiments. Various simple modifications can be made to the technical solutions of the present application within the scope of the technical idea of ​​the present application, and all of these simple modifications fall within the scope of protection of the present application.

[0139] It should be noted that the specific technical features described in the above specific embodiments may be combined in any appropriate manner as long as there is no contradiction, and in order to avoid unnecessary duplication, the present application does not separately describe various possible combination forms.

Claims

1. 8-(pyridylamide)-substituted coumarin compounds according to formula (I): 【Chemistry 1】 wherein ring A is independently selected from the group consisting of a phenyl group, a naphthyl group, and a 5- to 14-membered aromatic heterocyclic group; The phenyl group, naphthyl group and 5- to 14-membered aromatic heterocyclic group are unsubstituted or have 1 to 5 R ａ is replaced by R ａ each independently represents deuterium, a halogen, a hydroxyl group, a mercapto group, an amino group, a cyano group, a nitro group, an azido group, a methylsulfonyl group, an isopropylsulfonyl group, a benzenesulfonyl group, an aminosulfonyl group, a methanesulfonate group, an isopropanesulfonate group, a benzenesulfonate group, a trifluoromethyl group, a trifluoromethyloxy group, a formylamino group, C １～８ Alkyloxy group, C １～８ Alkylcarbonyl group, C １～８ Alkyloxycarbonyl group, C １～８ Alkyl group, C ３～８ selected from the group consisting of cycloalkyl groups and phenyl groups; The R ａ In the above, １～８ Alkyl group, C ３～８ The cycloalkyl group and the phenyl group are unsubstituted or have 1 to 5 R ｂ and R ｂ each independently represents deuterium, a halogen, a hydroxyl group, a mercapto group, an amino group, a cyano group, a nitro group, an azido group, a methylsulfonyl group, an isopropylsulfonyl group, a benzenesulfonyl group, an aminosulfonyl group, a methanesulfonate group, an isopropanesulfonate group, a benzenesulfonyl group, a fluoromethyl group, a trifluoromethyloxy group, a formylamino group, and １～８ alkylcarbonyl groups, 8-(pyridylamide) substituted coumarin compounds.

2. In formula (I), ring A is independently selected from the group consisting of a phenyl group, a naphthyl group, and a 5- to 14-membered aromatic heterocyclic group; The phenyl group, naphthyl group and 5- to 14-membered aromatic heterocyclic group are unsubstituted or have 1 to 5 R ａ is replaced by R ａ each independently represents deuterium, a halogen, a hydroxyl group, a mercapto group, an amino group, a cyano group, a nitro group, an azido group, a methylsulfonyl group, an isopropylsulfonyl group, a benzenesulfonyl group, an aminosulfonyl group, a methanesulfonate group, an isopropanesulfonate group, a benzenesulfonate group, a trifluoromethyl group, a trifluoromethyloxy group, a formylamino group, C １～６ Alkyloxy group, C １～６ Alkylcarbonyl group, C １～６ Alkyloxycarbonyl group, C １～６ Alkyl group, C ３～６ cycloalkyl groups, and phenyl groups; The R ａ In the above, １～６ Alkyl group, C ３～６ The cycloalkyl group and the phenyl group are unsubstituted or have 1 to 5 R ｂ and R ｂ are each independently deuterium, a halogen, a hydroxyl group, a mercapto group, an amino group, a cyano group, a nitro group, an azido group, a methylsulfonyl group, an isopropylsulfonyl group, a benzenesulfonyl group, an aminosulfonyl group, a methanesulfonate group, an isopropanesulfonate group, a benzenesulfonate group, a trifluoromethyl group, a trifluoromethyloxy group, a formylamino group, and C １～６ alkylcarbonyl groups, The 8-(pyridylamide) substituted coumarin compound according to claim 1.

3. The structure of the 8-(pyridylamide) substituted coumarin compound is shown in formula (IA): 【Chemistry 2】 However, R １ , R ２ , R ３ , R ４ and R ５ are independently hydrogen, deuterium, a halogen, a hydroxyl group, a mercapto group, an amino group, a cyano group, a nitro group, an azido group, a methylsulfonyl group, an isopropylsulfonyl group, a benzenesulfonyl group, an aminosulfonyl group, a methanesulfonate group, an isopropanesulfonate group, a benzenesulfonate group, a trifluoromethyl group, a trifluoromethyloxy group, a formylamino group, C １～４ Alkyloxy group, C １～４ Alkylcarbonyl group, C １～４ Alkyloxycarbonyl group, C １～４ selected from the group consisting of alkyl groups, and phenyl groups; 3. The 8-(pyridylamide) substituted coumarin compound according to claim 1 or 2.

4. The structure of the 8-(pyridylamide) substituted coumarin compound is shown in formula (IB): 【Chemistry 3】 However, R １ , R ２ , R ３ , R ４ and R ５ are independently hydrogen, deuterium, halogen, a hydroxyl group, a mercapto group, an amino group, a cyano group, a nitro group, a methylsulfonyl group, a trifluoromethyl group, a trifluoromethyloxy group, a formylamino group, C １～４ Alkyloxy group, C １～４ Alkylcarbonyl group, and C １～４ alkyl groups, and X is independently selected from N or C; 3. The 8-(pyridylamide) substituted coumarin compound according to claim 1 or 2.

5. The 8-(pyridylamide) substituted coumarin compound is selected from the following structures: 【Chemistry 4】 3. The 8-(pyridylamide) substituted coumarin compound according to claim 1 or 2.

6. A stereoisomer of the 8-(pyridylamide)-substituted coumarin compound according to any one of claims 1 to 5, a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition comprising the same, The pharmaceutical composition further comprises a pharma- ceutically acceptable adjuvant. A stereoisomer of an 8-(pyridylamide) substituted coumarin compound, a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition comprising the same. 【Request 7】 【Chemical 5】 and N,N-diisopropylethylamine, and then mixing and reacting with 4-(2-oxo-2H-chromene-8-carboxamido)pyridine-2-sulfonyl chloride to obtain a compound according to the present invention, wherein the range of the A ring is the same as that of any one of claims 1 to 5; The reaction formula is shown below: 【Chemistry 6】 A process for preparing the 8-(pyridylamide) substituted coumarin compounds according to any one of claims 1 to 5.

8. The method for preparing the 4-(2-oxo-2H-chromene-8-carboxamido)pyridine-2-sulfonyl chloride comprises the steps of: (1) Mix 8-bromocoumarin and n-butyllithium, then CO ２ mixing with the gas and reacting to obtain coumarin-8-carboxylic acid; (2) mixing coumarin-8-carboxylic acid with 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate and N,N-diisopropylethylamine, and then mixing with 4-aminopyridine-2-sulfonic acid to react to obtain 4-(2-oxo-2H-chromene-8-carboxamide)pyridine-2-sulfonic acid; (3) mixing 4-(2-oxo-2H-chromene-8-carboxamido)pyridine-2-sulfonic acid with dichlorosulfoxide and reacting to obtain 4-(2-oxo-2H-chromene-8-carboxamido)pyridine-2-sulfonyl chloride; The reaction formula is shown below: 【Chemistry 7】 A method for preparing the 8-(pyridylamide) substituted coumarin compounds according to claim 7.

9. The 8-(pyridylamide)-substituted coumarin compound according to any one of claims 1 to 5. Or use of a stereoisomer of the 8-(pyridylamide) substituted coumarin compound according to claim 6, a pharma- ceutical acceptable salt thereof, or a pharmaceutical composition containing the same in the preparation of a medicament for treating and / or preventing a disease or pathology associated with SIRT2 overactivity or overexpression of SIRT2.

10. The disease or condition associated with SIRT2 overactivity or overexpression includes Parkinson's disease, metabolic disease or tumor; 10. The use according to claim 9.

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