CN113149971A - 8- (indoletriazole) substituted coumarin compound and preparation method and application thereof - Google Patents

Abstract

The invention relates to an 8- (indole triazole) substituted coumarin compound and a preparation method and application thereof, wherein the structure of the 8- (indole triazole) substituted coumarin compound is shown as a formula (I), wherein a ring A is independently selected from phenyl, naphthyl and 5-14-membered aromatic heterocyclic group. The 8- (indole triazole) substituted coumarin compound related by the invention is a brand new compound structure and has stronger SIRT2 inhibitory activity, wherein the total content of 18 compoundsIn vitro SIRT2 inhibitory Activity IC₅₀Reach micromolar level and have obvious protective effect on neuroma cells. Therefore, the compound can be widely used for preparing medicaments for treating and/or preventing diseases or symptoms related to the SIRT2 high activity or SIRT2 overexpression or preparing medicaments for treating and/or preventing Parkinson's disease, metabolic diseases and tumors.

Description

8- (indoletriazole) substituted coumarin compound and preparation method and application thereof

Technical Field

The invention belongs to the technical field of medicines, and particularly relates to an 8- (indole triazole) substituted coumarin compound, a preparation method and application thereof, and in particular relates to an 8- (indole triazole) substituted coumarin compound capable of inhibiting SIRT2 activity or expression, and a preparation method and application thereof.

Background

Parkinson's disease is a common neurodegenerative disease, and its clinical manifestations are patient's bradykinesia, muscular rigidity and resting tremor. The pathological changes of parkinson's disease are mainly manifested in degenerative necrosis of mesocerebral substantia nigra dopaminergic neurons, and thereby in a significant reduction of striatal dopamine content. Patients with parkinson's disease are under a heavy physical and psychological burden. However, no medicament can play a role in protecting the degenerative diseases of nerve cells of the Parkinson disease at present. The therapeutic drugs for Parkinson disease (such as levodopa) on the market mainly relieve the symptoms of the disease, and the long-term administration of the drugs brings serious side effects to patients. Therefore, the search for new Parkinson disease treatment methods and the discovery of new Parkinson disease treatment drugs have very important significance.

As an important member of the Sirtuin family, SIRT2 is distributed mainly in the cytoplasm and its action substrate proteins include α -tubulin, histone H4, p53, FOXO and 14-3-3 protein. Functional studies of SIRT2 showed that: SIRT2 regulates the progression of cell mitosis by deacetylating H4-K16; SIRT2 maintains the stability of the genome by activating the activity of the APC/C system; SIRT2 mediates the mutual binding of Receptor-interacting proteins 1 and 3(RIP1 and RIP3) to regulate the progression of cell necrosis.

Recent studies have shown that: the activity of SIRT2 inhibition has potential value in the aspect of treating Parkinson's disease, and is particularly reflected in that: (1) SIRT2 is expressed in high abundance in the central nervous system of adult brain and regulates related physiological metabolism; (2) siRNA of SIRT2 and a small molecule inhibitor AGK2 can save nerve cell toxicity caused by alpha-synulein, and AGK2 can also protect nerve cells of a transgenic drosophila parkinsonii model in vivo in a dose-dependent manner to reduce apoptosis; (3) SIRT2 increased Bim expression at the RNA and protein levels by deacetylating FOXO3a, promoting apoptosis in a parkinson's disease cell model of MPTP action and in a mouse model brain of MPTP action. However, the phenomenon of significantly reducing nigrostriatal damage was shown in the brains of SIRT2 knock-out mice; (4) in addition, the small molecule inhibitor AK-7 of SIRT2 can obviously reduce the aggregation of mutant Huntingtin in the brain of a transgenic mouse with Huntington disease, another neurodegenerative disease, and can obviously improve the behavior of the transgenic mouse with Huntington disease and prolong the survival time of the transgenic mouse.

Representative prior art SIRT2 small molecule inhibitors include the following:

however, the types of the SIRT2 inhibitors known in the prior art are quite limited, and in view of the potential activity of the SIRT2 small-molecule inhibitor in treating Parkinson's disease, the design and synthesis of a novel SIRT2 inhibitor are very practical.

Disclosure of Invention

Aiming at the defects of the prior art, the invention aims to provide an 8- (indole triazole) substituted coumarin compound and a preparation method and application thereof, and particularly provides an 8- (indole triazole) substituted coumarin compound capable of inhibiting SIRT2 activity or expression and a preparation method and application thereof. The invention develops a 8- (indole triazole) substituted coumarin compound with a novel structure, has obvious SIRT2 inhibitory activity and better selectivity on SIRT2, and can be used for preventing and treating Parkinson's disease, metabolic diseases, tumors and the like.

In order to achieve the purpose, the invention adopts the following technical scheme:

in a first aspect, the invention provides an 8- (indole triazole) substituted coumarin compound, wherein the structure of the 8- (indole triazole) substituted coumarin compound is shown as the formula (I):

wherein the a ring is independently selected from phenyl, naphthyl, 5-14 membered aromatic heterocyclic group (e.g., 5 membered aromatic heterocyclic group, 6 membered aromatic heterocyclic group, 7 membered aromatic heterocyclic group, 8 membered aromatic heterocyclic group, 9 membered aromatic heterocyclic group, 10 membered aromatic heterocyclic group, 11 membered aromatic heterocyclic group, 12 membered aromatic heterocyclic group, 13 membered aromatic heterocyclic group, 14 membered aromatic heterocyclic group);

said phenyl, naphthyl, 5-14 membered aromatic heterocyclic group being unsubstituted or substituted by 1-5 (e.g. 1,2,3, 4, 5) R_aSubstitution; r_aEach independently selected from deuterium, halogen, hydroxy, mercapto, amino, cyano, nitro, azido, methanesulfonyl, isopropylsulfonyl, benzenesulfonyl, aminosulfonyl, methanesulfonate, isopropylsulfonate, benzenesulfonate, trifluoromethyl, trifluoromethyloxy, carboxamido, C_1-8Alkyloxy, C_1-8Alkylcarbonyl group, C_1-8Alkyl oxycarbonyl radical, C_1-8Alkyl radical, C_3-8Cycloalkyl, phenyl;

in said R_aIn (A), the C_1-8Alkyl radical, C_3-8Cycloalkyl, phenyl unsubstituted or substituted by 1-5 (e.g. 1,2,3, 4, 5) R_bSubstitution; the R is_bEach independently selected from deuterium, halogen, hydroxy, mercapto, amino, cyano, nitro, azido, methanesulfonyl, isopropylsulfonyl, benzenesulfonyl, aminosulfonyl, methanesulfonate, isopropylsulfonate, benzenesulfonate, trifluoromethyl, trifluoromethyloxy, carboxamido, C_1-8An alkylcarbonyl group.

C above_1-8Means that the number of carbon atoms of the substituent is 1,2,3, 4, 5, 6, 7 or 8; c_3-8The number of carbon atoms of the substituent is 3,4, 5, 6, 7 or 8.

The 8- (indole triazole) substituted coumarin compound related by the invention is a brand-new compound structure, the in vitro SIRT2 inhibitory activity of the compound reaches more than 50%, wherein the in vitro SIRT2 inhibitory activity IC of 18 compounds₅₀To micromolar levels, in particularIs IC of 13 of the compounds₅₀Value of up to 10^-7mol/L level, wherein IC of 3 compounds₅₀Value of up to 10^-8At the mol/L level, the compound shows good SIRT2 inhibition activity and has a remarkable protection effect on neuroma cells. Therefore, the compound can be widely used for preparing medicaments for treating and/or preventing diseases or symptoms related to the SIRT2 high activity or SIRT2 overexpression or preparing medicaments for treating and/or preventing Parkinson's disease, metabolic diseases and tumors.

Preferably, in formula (I), the a ring is independently selected from phenyl, naphthyl, 5-14 membered aromatic heterocyclic group (e.g., 5 membered aromatic heterocyclic group, 6 membered aromatic heterocyclic group, 7 membered aromatic heterocyclic group, 8 membered aromatic heterocyclic group, 9 membered aromatic heterocyclic group, 10 membered aromatic heterocyclic group, 11 membered aromatic heterocyclic group, 12 membered aromatic heterocyclic group, 13 membered aromatic heterocyclic group, 14 membered aromatic heterocyclic group);

said phenyl, naphthyl, 5-14 membered aromatic heterocyclic group being unsubstituted or substituted by 1-5 (e.g. 1,2,3, 4, 5) R_aSubstitution; r_aEach independently selected from deuterium, halogen, hydroxy, mercapto, amino, cyano, nitro, azido, methanesulfonyl, isopropylsulfonyl, benzenesulfonyl, aminosulfonyl, methanesulfonate, isopropylsulfonate, benzenesulfonate, trifluoromethyl, trifluoromethyloxy, carboxamido, C_1-6Alkyloxy, C_1-6Alkylcarbonyl group, C_1-6Alkyl oxycarbonyl radical, C_1-6Alkyl radical, C_3-6Cycloalkyl, phenyl;

in said R_aIn (A), the C_1-6Alkyl radical, C_3-6Cycloalkyl, phenyl unsubstituted or substituted by 1-5 (e.g. 1,2,3, 4, 5) R_bSubstitution; the R is_bEach independently selected from deuterium, halogen, hydroxy, mercapto, amino, cyano, nitro, azido, methanesulfonyl, isopropylsulfonyl, benzenesulfonyl, aminosulfonyl, methanesulfonate, isopropylsulfonate, benzenesulfonate, trifluoromethyl, trifluoromethyloxy, carboxamido, C_1-6An alkylcarbonyl group.

C above_1-6Means that the number of carbon atoms of the substituent is 1,2,3, 4, 5 or 6; c_3-6The number of carbon atoms of the substituent is 3,4, 5 or 6.

Preferably, the structure of the 8- (indole triazole) substituted coumarin compound is shown as the formula (IA):

wherein R is₁、R₂、R₃、R₄、R₅Independently selected from hydrogen, deuterium, halogen, hydroxy, mercapto, amino, cyano, nitro, azido, methanesulfonyl, isopropylsulfonyl, benzenesulfonyl, aminosulfonyl, methanesulfonate, isopropylsulfonate, benzenesulfonate, trifluoromethyl, trifluoromethyloxy, carboxamido, C_1-4Alkyloxy, C_1-4Alkylcarbonyl group, C_1-4Alkyl oxycarbonyl radical, C_1-4Alkyl, phenyl.

C above_1-4The number of carbon atoms of the substituent is 1,2,3 or 4.

Preferably, the structure of the 8- (indole triazole) substituted coumarin compound is shown as the formula (IB):

wherein R is₁、R₂、R₃、R₄、R₅Independently selected from hydrogen, deuterium, halogen, hydroxy, mercapto, amino, cyano, nitro, methylsulfonyl, trifluoromethyl, trifluoromethyloxy, carboxamido, C_1-4Alkyloxy, C_1-4Alkylcarbonyl group, C_1-4An alkyl group; x is independently selected from N or C.

C above_1-4The number of carbon atoms of the substituent is 1,2,3 or 4.

Further, the 8- (indole triazole) substituted coumarin compound is selected from structures (represented by structural formulas or system names respectively) shown as follows:

in a second aspect, the invention provides a stereoisomer of the 8- (indoletriazole) substituted coumarin compound, a pharmaceutically acceptable salt thereof or a pharmaceutical composition containing the stereoisomer;

preferably, the pharmaceutical composition further comprises pharmaceutically acceptable pharmaceutical excipients, such as carriers, diluents, excipients, fillers, binders, wetting agents, disintegrants, emulsifiers, cosolvents, solubilizers, osmotic pressure regulators, surfactants, coating materials, colorants, pH regulators, antioxidants, bacteriostats, buffers, or the like.

The pharmaceutically acceptable salt of the 8- (indole triazole) substituted coumarin compound related by the invention is a salt formed by the 8- (indole triazole) substituted coumarin compound and acid selected from the following acids: hydrochloric acid, hydrobromic acid, p-toluenesulfonic acid, tartaric acid, maleic acid, lactic acid, methanesulfonic acid, sulfuric acid, phosphoric acid, citric acid, acetic acid or trifluoroacetic acid. Preferably hydrochloric acid, hydrobromic acid, p-toluenesulfonic acid or trifluoroacetic acid.

In a third aspect, the invention provides a preparation method of the 8- (indole triazole) substituted coumarin compound, which comprises the following steps:

(1) will be provided with

Mixing with 7- (1- (1- (2-oxo) -2H-chromen-8-yl) -1H-1,2, 3-triazol-4-yl) -1H-indole-1-carboxylic acid tert-butyl ester, and reacting to obtain an intermediate product, namely 2- (4-chlorobenzyl) -7- (1- (2-oxo-2H-chromen-8-yl) -1H-1,2, 3-triazol-4-yl) -1H-indole-1-carboxylic acid tert-butyl ester;

(2) hydrolyzing 2- (4-chlorobenzyl) -7- (1- (2-oxo-2H-chromen-8-yl) -1H-1,2, 3-triazole-4-yl) -1H-indole-1-tert-butyl formate under acidic condition, and adjusting pH to neutrality to obtain the final product;

wherein the defined range of the a-ring is in accordance with the range defined in the first aspect;

the reaction formula is shown as follows:

preferably, the step (1) is to

Before mixing with tert-butyl 7- (1- (1- (2-oxo) -2H-chromen-8-yl) -1H-1,2, 3-triazol-4-yl) -1H-indole-1-carboxylate, tert-butyl 7- (1- (1- (2-oxo) -2H-chromen-8-yl) -1H-1,2, 3-triazol-4-yl) -1H-indole-1-carboxylate is sequentially mixed and stirred with n-butyl lithium and tributyltin chloride;

preferably, tris (dibenzylideneacetone) dipalladium, dicyclohexylphosphinobenzene, potassium fluoride and cesium carbonate are further added to the reaction system in the step (1).

Preferably, the preparation method of 7- (1- (1- (2-oxo) -2H-chromen-8-yl) -1H-1,2, 3-triazol-4-yl) -1H-indole-1-carboxylic acid tert-butyl ester in the step (1) comprises the following steps:

(1) reacting 8-bromocoumarin with NaN₃Mixing and reacting to obtain 8-azido coumarin;

(2) mixing 8-azido coumarin with 7-ethynyl-1H-indole and cuprous iodide, and reacting to obtain 8- (4- (1H-indol-7-yl) -1H-1,2, 3-triazol-1-yl) -2H-chromen-2-one;

(3) mixing 8- (4- (1H-indol-7-yl) -1H-1,2, 3-triazol-1-yl) -2H-chromen-2-one with sodium hydrideAnd then (Boc) was added thereto₂O to give 7- (1- (1- (2-oxo) -2H-chromen-8-yl) -1H-1,2, 3-triazol-4-yl) -1H-indole-1-carboxylic acid tert-butyl ester;

the reaction formula is shown as follows:

the preparation method of the 8- (pyridine triazole) substituted coumarin compound is simple and easy to operate, suitable for industrial production and practical.

As a preferred technical scheme of the invention, the preparation method of the 8- (indole triazole) substituted coumarin compound specifically comprises the following steps:

(1) addition of NaN to an ultra-dry DMF solution of 8-bromocoumarin at room temperature₃. The reaction solution was heated. The reaction was completed by TLC, and the reaction mixture was cooled to room temperature and quenched with saturated brine. The product was extracted three times with dichloromethane. And drying the combined organic phases by using anhydrous sodium sulfate, filtering, evaporating the solvent, and purifying by using a silica gel column to obtain the product 8-azido coumarin.

(2) 8-azidocoumarin, 7-ethynyl-1H-indole and cuprous iodide were added to ultra-dry dichloromethane and the reaction was stirred at room temperature. After TLC detection reaction is completed, the product 8- (4- (1H-indole-7-yl) -1H-1,2, 3-triazole-1-yl) -2H-chromen-2-ketone is obtained by purifying with a silica gel column.

(3) Adding sodium hydride to 8- (4- (1H-indole)-7-yl) -1H-1,2, 3-triazol-1-yl) -2H-chromen-2-one in anhydrous tetrahydrofuran. The reaction solution was stirred at room temperature. To the above solution was added (Boc) in portions with stirring₂And O. The reaction was stirred at room temperature and after completion of the TLC check, quenched with a mixture of ice and water. The mixture was extracted three times with ethyl acetate, and the organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated. Purifying with silica gel column to obtain product 7- (1- (1- (2-oxo-2H-chromen-8-yl) -1H-1,2, 3-triazole-4-yl) -1H-indole-1-tert-butyl formate.

(4) To a solution of tert-butyl 7- (1- (1- (2-oxo) -2H-chromen-8-yl) -1H-1,2, 3-triazol-4-yl) -1H-indole-1-carboxylate in super dry THF was added n-butyllithium and the mixture was stirred at 0 ℃. To the reaction solution was added an ultra-dry THF solution of tributyltin chloride, and the mixture was stirred at room temperature. Brine and ethyl acetate were added to the reaction solution, extracted three times, the organic phases were combined, dried over anhydrous sodium sulfate, filtered and concentrated to give an orange liquid. To the orange liquid and a solution of 1-chloro-4- (iodomethyl) benzene in 1, 4-dioxane was added tris (dibenzylideneacetone) dipalladium, dicyclohexylphosphinobenzene, potassium fluoride and cesium carbonate. The mixture was stirred under argon overnight. The reaction was checked by TLC, cooled to room temperature after completion of the reaction, then filtered through celite and washed with 1, 4-dioxane. Concentrating the filtrate, and purifying with silica gel column to obtain intermediate 2- (4-chlorobenzyl) -7- (1- (2-oxo-2H-chromen-8-yl) -1H-1,2, 3-triazole-4-yl) -1H-indole-1-tert-butyl formate.

(5) To a solution of hydrochloric acid in 1, 4-dioxane was added tert-butyl 2- (4-chlorobenzyl) -7- (1- (2-oxo-2H-chromen-8-yl) -1H-1,2, 3-triazol-4-yl) -1H-indole-1-carboxylate and stirred at room temperature. After concentration, water is added, and the pH value is adjusted to be neutral. Extracted three times with ethyl acetate, the organic phases are combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated. Purifying with silica gel column to obtain the product.

In a fourth aspect, the invention provides an application of the 8- (indole triazole) substituted coumarin compound according to the first aspect or the stereoisomer of the 8- (indole triazole) substituted coumarin compound according to the second aspect, a pharmaceutically acceptable salt thereof, and a pharmaceutical composition containing the stereoisomer in preparation of drugs for treating and/or preventing diseases or symptoms related to SIRT2 overhigh activity or SIRT2 overexpression.

Preferably, the disease or disorder associated with SIRT2 hyperactivity or SIRT2 overexpression includes parkinson's disease, metabolic disease, or tumor.

In a fifth aspect, the invention also provides an 8- (indole triazole) substituted coumarin compound in the first aspect or a stereoisomer of the 8- (indole triazole) substituted coumarin compound in the second aspect, a pharmaceutically acceptable salt thereof, and an application of a pharmaceutical composition containing the stereoisomer in preparation of a SIRT2 inhibitor.

In a sixth aspect, the invention also provides a method of treating and/or preventing a disease or condition associated with SIRT2 hyperactivity or SIRT2 overexpression, the method comprising: administering to a subject in need thereof a therapeutically and/or prophylactically effective amount of a stereoisomer of an 8- (indoletriazole) substituted coumarin as described in the first aspect or an 8- (indoletriazole) substituted coumarin as described in the second aspect, a pharmaceutically acceptable salt thereof, and a pharmaceutical composition comprising the same.

Preferably, the disease or disorder associated with SIRT2 hyperactivity or SIRT2 overexpression includes parkinson's disease, metabolic disease, or tumor.

Various aspects and features of the disclosure are described further below.

The various terms and phrases used herein have the ordinary meaning as is known to those skilled in the art, and even then, it is intended that the present invention not be limited to the specific meanings and expressions employed herein as are inconsistent with such known meanings. The following are definitions of various terms used herein, which apply to the terms used throughout the specification of the present application unless otherwise specified in specific instances. Definitions for various groups of the compounds of the present invention are provided below and, unless otherwise defined, are used uniformly throughout the specification and claims.

As referred to herein, the terms "halo", "halogen atom", "halo" and the like denote fluorine, chlorine, bromine or iodine, in particular fluorine, chlorine or bromine.

As referred to herein, the term "alkyl" refers to an alkyl group having the indicated number of carbon atoms, which may be straight or branched, such as the indicated "C_1-8The "alkyl group" means an alkyl group having 1,2,3, 4, 5, 6, 7 or 8 carbon atoms and may include C_1-8Alkyl radical, C_1-7Alkyl radical, C_2-8Alkyl radical, C_2-7Alkyl radical, C_2-6Alkyl radical, C_3-8Alkyl radical, C_3-7Alkyl radical, C_3-6Alkyl group, and the like, and preferable specific groups include methyl group, ethyl group, n-propyl group, isopropyl group, n-butyl group, sec-butyl group, tert-butyl group, pentyl group, hexyl group, heptyl group, and octyl group, and methyl group and isopropyl group are more preferable. For example said "C_1-6Alkyloxy group and C_1-6Alkyl formyl group and C_1-6Alkyl oxygen formyl radical "or" C_1-6C in alkyl_1-6Alkyl "refers to an alkyl group having 1,2,3, 4, 5, 6 carbon atoms, and may include C_1-5Alkyl radical, C_1-4Alkyl radical, C_2-6Alkyl radical, C_2-5Alkyl radical, C_2-4Alkyl radical, C_3-6Alkyl radical, C_3-5Alkyl radical, C_3-4Alkyl, etc., and preferable specific groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, pentyl, hexyl, and methyl is more preferable. For example said "C_1-4Alkylcarbonyl "or" C_1-4"C in" alkyloxycarbonyl_1-4Alkyl "refers to an alkyl group having 1,2,3, 4 carbon atoms, and may include C_1-4Alkyl radical, C_2-4Alkyl, etc., and preferred specific groups are methyl, ethyl, n-propyl, isopropyl.

As referred to herein, the term "cycloalkyl" refers to cyclic alkyl groups having the indicated number of ring carbon atoms, for example, where reference is made to "C3-8 cycloalkyl", this refers to cycloalkyl groups having 3,4, 5, 6, 7, 8 carbon atoms and may include a sub-range of groups represented by C3-7 cycloalkyl, C3-4 cycloalkyl, C4-6 cycloalkyl, and the like, as well as preferred specific groups such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, and further preferably cyclopropyl, cyclopentyl, cyclohexyl. For example, the "C3-6 cycloalkyl group" refers to a cycloalkyl group having 3,4, 5 or 6 carbon atoms, and may include groups in a sub-range represented by C3-6 cycloalkyl, C3-5 cycloalkyl, C4-5 cycloalkyl, and the like, and preferable specific groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and further preferable are cyclopropyl, cyclopentyl, and cyclohexyl.

As referred to herein, the term "arylheterocyclyl" refers to a heterocyclic aromatic system containing 1 to 4 heteroatoms, including heteroatoms of nitrogen, oxygen and sulfur. As referred to herein, "5-14 membered aromatic heterocyclyl" refers to a heterocyclic aromatic system containing 5-14 ring atoms. Specific examples include aryl groups containing 1 carbon atom and 4 heteroatoms selected from nitrogen, oxygen, sulfur, preferred specific groups such as tetrazolyl; aryl containing 2 carbon atoms and 3 heteroatoms selected from nitrogen, oxygen, sulfur, preferred specific groups such as 1,2, 3-triazolyl, 1,2, 4-triazolyl, oxadiazolyl, thiadiazolyl; aryl containing 3 carbon atoms and 2 heteroatoms selected from nitrogen, oxygen, sulfur, preferred specific groups such as imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl; aryl containing 4 carbon atoms and 1-2 heteroatoms selected from nitrogen, oxygen, sulfur, with preferred embodiments such as pyrrolyl, furyl, thienyl, pyridazinyl, pyrimidinyl, pyrazinyl; aryl containing 5 carbon atoms and 1 heteroatom selected from nitrogen, oxygen, sulfur, preferred embodiments are e.g. pyridyl, preferably pyridyl; aryl containing 6 carbon atoms and 3 heteroatoms selected from nitrogen, oxygen, sulfur, preferred embodiments are groups such as benzotriazolyl; aryl containing 7 carbon atoms and 2 heteroatoms selected from nitrogen, oxygen, sulfur, preferred embodiments are benzimidazolyl, benzpyrazolyl; aryl containing 8 carbon atoms and 1-2 heteroatoms selected from nitrogen, oxygen, sulfur, with preferred embodiments such as indolyl, benzofuranyl, benzothienyl, benzopyrazinyl, benzopyrimidinyl, and benzopyrazinyl; aryl containing 9 carbon atoms and 1 heteroatom selected from nitrogen, oxygen, sulfur, with particular preference given to quinolinyl, isoquinolinyl.

As referred to herein, the term "effective amount" refers to a dose that achieves treatment and/or prevention of a disease or disorder described herein in a subject.

As referred to herein, the term "pharmaceutical composition" may also refer to a "composition" which may be used to effect treatment and/or prevention of a disease or disorder described herein in a subject, particularly a mammal.

As referred to herein, the term "subject" may refer to a patient or other animal, particularly a mammal, e.g., a human, dog, monkey, cow, horse, etc., receiving a compound, pharmaceutically acceptable salt or pharmaceutical composition thereof referred to herein for the treatment and/or prevention of a disease or condition described herein.

As referred to herein, the term "disease and/or disorder" refers to a physical condition of the subject that is associated with the disease and/or disorder of the present invention. For example, the disease and/or condition of the present invention may refer to either a physical state, such as that which is indicative of Parkinson's disease, or a disease state, such as that which is indicative of Parkinson's disease. The body state and the disease state are not distinguished herein or may be referred to one another, e.g., "parkinson's disease" may be used interchangeably with "parkinson's disease".

As referred to herein, the term "pharmaceutically acceptable" when describing a "pharmaceutically acceptable salt" means not only that the salt is physiologically acceptable to the subject, but may also refer to a synthetic substance of pharmaceutical value, e.g., a salt formed as an intermediate in order to effect chiral resolution, which salt may play a role in obtaining the final product of the invention, although such intermediate salt may not be directly administered to the subject.

In a further aspect, the invention relates to pharmaceutical compositions comprising as active ingredient a compound of the invention. The pharmaceutical composition may be prepared according to methods well known in the art. The compounds of the invention may be formulated into any dosage form suitable for human or animal use by combining them with one or more pharmaceutically acceptable solid or liquid excipients and/or adjuvants. The compounds of the present invention are generally present in the pharmaceutical compositions in an amount of from 0.1 to 95% by weight.

The compounds of the present invention or pharmaceutical compositions containing them may be administered in unit dosage form by enteral or parenteral routes, such as oral, intravenous, intramuscular, subcutaneous, nasal, oromucosal, ophthalmic, pulmonary and respiratory, dermal, vaginal, rectal, and the like.

The dosage form for administration may be a liquid dosage form, a solid dosage form, or a semi-solid dosage form. The liquid dosage forms can be solution (including true solution and colloidal solution), emulsion (including o/w type, w/o type and multiple emulsion), suspension, injection (including water injection, powder injection and infusion), eye drop, nose drop, lotion, liniment, etc.; the solid dosage form can be tablet (including common tablet, enteric coated tablet, buccal tablet, dispersible tablet, chewable tablet, effervescent tablet, orally disintegrating tablet), capsule (including hard capsule, soft capsule, and enteric coated capsule), granule, powder, pellet, dripping pill, suppository, pellicle, patch, aerosol (powder), spray, etc.; semisolid dosage forms can be ointments, gels, pastes, and the like.

The compound can be prepared into common preparations, sustained release preparations, controlled release preparations, targeting preparations and various particle delivery systems.

For tableting the compound of the present invention, a wide variety of excipients known in the art may be used, including diluents, binders, wetting agents, disintegrants, lubricants, and solubilizers. The diluent can be starch, dextrin, sucrose, glucose, lactose, mannitol, sorbitol, xylitol, microcrystalline cellulose, calcium sulfate, calcium hydrogen phosphate, calcium carbonate, etc.; the humectant can be water, ethanol, isopropanol, etc.; the binder can be starch slurry, dextrin, syrup, Mel, glucose solution, microcrystalline cellulose, acacia slurry, gelatin slurry, sodium carboxymethylcellulose, methylcellulose, hydroxypropyl methylcellulose, ethyl cellulose, acrylic resin, carbomer, polyvinylpyrrolidone, polyethylene glycol, etc.; the disintegrant may be dry starch, microcrystalline cellulose, low-substituted hydroxypropyl cellulose, crosslinked polyvinylpyrrolidone, crosslinked sodium carboxymethylcellulose, sodium carboxymethyl starch, sodium bicarbonate and citric acid, polyoxyethylene sorbitol fatty acid ester, sodium dodecyl sulfate, etc.; the lubricant and cosolvent may be talc, silica, stearate, tartaric acid, liquid paraffin, polyethylene glycol, etc.

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

To encapsulate the administration unit, the active ingredient of the compound of the present invention may be mixed with a diluent and a cosolvent, and the mixture may be directly placed in a hard capsule or soft capsule. Or the effective component of the compound of the invention can be prepared into granules or pellets with diluent, adhesive and disintegrating agent, and then placed into hard capsules or soft capsules. The diluents, binders, wetting agents, disintegrants, and cosolvents used to prepare the compound tablets of the present invention can also be used to prepare capsules of the compounds of the present invention.

For preparing the compound of the present invention into injection, water, ethanol, isopropanol, propylene glycol or their mixture can be used as solvent, and appropriate amount of solubilizer, cosolvent, pH regulator, and osmotic pressure regulator commonly used in the art can be added. The solubilizer or cosolvent can be poloxamer, lecithin, hydroxypropyl-beta-cyclodextrin, etc.; the pH regulator can be phosphate, acetate, hydrochloric acid, sodium hydroxide, etc.; the osmotic pressure regulator can be sodium chloride, mannitol, glucose, phosphate, acetate, etc. For example, mannitol and glucose can be added as proppant for preparing lyophilized powder for injection.

In addition, colorants, preservatives, flavors, or other additives may also be added to the pharmaceutical preparation, if desired.

For the purpose of administration and enhancing the therapeutic effect, the drug or pharmaceutical composition of the present invention can be administered by any known administration method.

The dosage of the pharmaceutical composition of the compound of the present invention to be administered may 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 route and dosage form of administration, and the like. Generally, a suitable daily dosage range for a compound of the invention is from 0.001 to 150mg/kg body weight, preferably from 0.1 to 100mg/kg body weight, more preferably from 1 to 60mg/kg body weight, most preferably from 2 to 30mg/kg body weight. The above-described dosage may be administered in one dosage unit or divided into several dosage units, depending on the clinical experience of the physician and the dosage regimen including the use of other therapeutic means.

The compounds or compositions of the present invention may be administered alone or in combination with other therapeutic or symptomatic agents. When the compound of the present invention is used in a synergistic manner with other therapeutic agents, the dosage thereof should be adjusted according to the actual circumstances.

Compared with the prior art, the invention has the following beneficial effects:

the 8- (indole triazole) substituted coumarin compound related by the invention is a brand-new compound structure, the in vitro SIRT2 inhibitory activity of the compound reaches more than 50%, wherein the in vitro SIRT2 inhibitory activity IC of 18 compounds₅₀To micromolar levels, especially IC of 13 compounds therein₅₀Value of up to 10^-7mol/L level, wherein IC of 3 compounds₅₀Value of up to 10^-8At the mol/L level, the compound shows good SIRT2 inhibition activity and has a remarkable protection effect on neuroma cells. Therefore, the compound can be widely used for preparing medicaments for treating and/or preventing diseases or symptoms related to the SIRT2 high activity or SIRT2 overexpression or preparing medicaments for treating and/or preventing Parkinson's disease, metabolic diseases and tumors.

Drawings

FIG. 1 is a statistical chart of the protection effect of 8- (indoletriazole) substituted coumarin compounds on SH-SY5Y cell damage in test example 3.

Detailed Description

The technical solution of the present invention is further explained by the following embodiments. It should be understood by those skilled in the art that the examples are only for the understanding of the present invention and should not be construed as the specific limitations of the present invention.

For all of the following examples or preparations, standard procedures and purification methods known to those skilled in the art may be used. Unless otherwise indicated, all temperatures are in degrees celsius (degrees celsius) and the structure of a compound is determined by nuclear magnetic resonance spectroscopy (NMR) and/or Mass Spectroscopy (MS).

For all the following examples or preparations, the structure of the compounds is determined by NMR¹H NMR) or Mass Spectrometry (MS). The nuclear magnetic resonance hydrogen spectral shift (δ) is given in parts per million (ppm). The NMR spectra were measured using a NMR spectrometer model Mercury-300 or Mercury-400, deuterated chloroform (CDCl3) or deuterated dimethyl sulfoxide (DMSO-d6) as solvent, and Tetramethylsilane (TMS) or 3- (trimethylsilyl) deuterated sodium propionate (TSM) as internal standard.

The electronic balance used was an electronic balance model Yanaco LY-300, Japan.

The column chromatography uses 200-300 mesh or 300-400 mesh silica gel as a carrier.

The anhydrous solvents were all processed by standard methods. Other reagents were all commercially available analytical grade.

Wherein the content of the first and second substances,

HATU is 2- (7-Azabenzotriazol-1-yl) -N, N, N ', N' -tetramethyluronium hexafluoro-phosphate, i.e., 2- (7-azabenzotriazole) -N, N, N ', N' -tetramethyluronium hexafluorophosphate.

DIPEA is N, N-diisomyricylethylamine, i.e., N-diisopropylethylamine.

DMF is N, N-dimethylformamide, i.e., N-dimethylformamide.

The following preparations and examples refer to the following sources of the reaction raw materials:

8-bromocoumarin is available from Shanghai Aladdin Biotechnology GmbH, CAS: 33491-30-4;

7-ethynyl-1H-indole is available from Shanghai Wo, Biotech, Inc.;

1-chloro-4- (iodomethyl) benzene, 1-chloro-3- (iodomethyl) benzene, 1-chloro-2- (iodomethyl) benzene, 1-methyl-4- (iodomethyl) benzene, 1-nitro-3- (iodomethyl) benzene, 1-cyano-2- (iodomethyl) benzene, 1-fluoro-4- (iodomethyl) benzene, 1-methanesulfonyl-4- (iodomethyl) benzene, 1-trifluoromethyl-4- (iodomethyl) benzene, 1-phenyl-4- (iodomethyl) benzene, 1-hydroxy-4- (iodomethyl) benzene, 1-methoxy-4- (iodomethyl) benzene, 1, 2-fluoro-4- (iodomethyl) benzene, methyl-1-hydroxy-4- (iodomethyl) benzene, methyl-1-methyl-4- (iodomethyl) benzene, methyl-1, 2-fluoro-4- (iodomethyl) benzene, methyl-1, 2-methyl-4- (iodomethyl) benzene, and mixtures thereof, 1-chloro-2-bromo-4- (iodomethyl) benzene, 2-bromo-8- (iodomethyl) naphthalene, 2-chloro-4- (iodomethyl) naphthalene, 5- (iodomethyl) quinoline, and 3-bromo-5- (iodomethyl) quinoline are available from Shanghai Michelin Biotechnology, Inc., Sigma-Aldrich Sigma Aldrich (Shanghai) trade, Beijing Bailingwei technology, Shanghai Michelle chemical, Inc., Afaha chemical, Inc., Beijing coupled technology, Shanghai Arlatin Biotechnology, Inc., Beijing Sorbolea Tech technology, Inc., and Beijing Yinokay, Inc., respectively.

Preparation example 1

This preparation example prepared intermediate 4, shown below, via the following synthetic route:

(1) to an ultra-dry DMF solution (15mL) of 8-bromocoumarin (5.00g, 22.33mmol) at 20 deg.C was added NaN3(1.74g, 26.80 mmol). The reaction solution was heated at 40 ℃ for 14 hours. The completion of the reaction was checked by TLC, and the reaction mixture was cooled to 20 ℃ and quenched with saturated brine. The product was extracted three times with dichloromethane (20 mL. times.3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and then the solvent was evaporated and purified by silica gel column to obtain the product 8-azidocoumarin (1.38g, yield 36%).

(2) 8-azidocoumarin (1.30g, 6.95mmol), 7-ethynyl-1H-indole (0.98g, 6.95mmol) and cuprous iodide (0.13g, 0.70mmol) were added to ultra-dry dichloromethane (15mL) and the reaction was stirred at 20 ℃ for 25 min. After TLC detection reaction, the product 8- (4- (1H-indol-7-yl) -1H-1,2, 3-triazol-1-yl) -2H-chromen-2-one (1.94g, 85% yield) was obtained by purification on silica gel column.

(3) Sodium hydride (0.16g, 6.59mmol) was added to a solution of 8- (4- (1H-indol-7-yl) -1H-1,2, 3-triazol-1-yl) -2H-chromen-2-one (1.80g, 5.49mmol) in anhydrous tetrahydrofuran (30mL) at-10 ℃. The reaction solution was stirred at 20 ℃ for 1 hour. To the above solution was added (Boc) in portions with stirring₂O (1.44g, 6.59 mmol). The reaction was stirred at 20 ℃ for 4 hours and after completion of the TLC check, it was quenched with an ice-water mixture (20 g). The mixture was extracted three times with ethyl acetate (20mL × 3), and the organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated. Purification on silica gel column afforded the product, tert-butyl 7- (1- (1- (2-oxo) -2H-chromen-8-yl) -1H-1,2, 3-triazol-4-yl) -1H-indole-1-carboxylate (1.88g, yield 80%).

Example 1

This example prepares compound 1: 8- (4- (2- (4-chlorobenzyl) -1H-indol-7-yl) -1H-1,2, 3-triazole-1-yl) -2H-chromen-2-one, the structure of which is shown as follows:

(1) to a cold (0 ℃) solution of tert-butyl 7- (1- (1- (2-oxo) -2H-chromen-8-yl) -1H-1,2, 3-triazol-4-yl) -1H-indole-1-carboxylate (1.80g, 4.20mmol) in super dry THF (20mL) was added n-butyllithium (1.60M in n-hexane; 2.63mL, 4.20mmol) and the mixture was stirred at 0 ℃ for 1 hour. To the reaction solution was added a solution of tributyltin chloride (1.37g, 4.20mmol) in ultra dry THF (10mL), and the mixture was stirred at 20 ℃ for 1.5 hours. Brine (10mL) and ethyl acetate (20mL) were added to the reaction solution, extracted three times, and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to give an orange liquid. To a solution of this orange liquid (3.00mmol) and 1-chloro-4- (iodomethyl) benzene (0.76g, 3.00mmol) in 1, 4-dioxane (10mL) was added tris (dibenzylideneacetone) dipalladium (0.17g, 0.30mmol), dicyclohexylphosphinobenzene (0.26g, 0.75mmol), potassium fluoride (0.52g, 9.00mmol) and cesium carbonate (1.95g, 6.00 mmol). The mixture was stirred at 60 ℃ for 24h under argon. The reaction was checked by TLC, cooled to 20 ℃ after completion of the reaction, filtered through celite and washed with 1, 4-dioxane (10 mL). Concentrating the filtrate, and purifying with silica gel column to obtain intermediate 2- (4-chlorobenzyl) -7- (1- (2-oxo-2H-chromen-8-yl) -1H-1,2, 3-triazole-4-yl) -1H-indole-1-tert-butyl formate.

(2) To a solution of hydrochloric acid (4.0M) in 1, 4-dioxane (10mL) was added tert-butyl 2- (4-chlorobenzyl) -7- (1- (2-oxo-2H-chromen-8-yl) -1H-1,2, 3-triazol-4-yl) -1H-indole-1-carboxylate (0.5g, 0.91mmol) and stirred at 20 ℃ for 3 hours. After concentration, water was added, and the pH was adjusted to neutral with 1N aqueous sodium hydroxide solution. Extracted three times with ethyl acetate (20 mL. times.3), the organic phases are combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated. Purifying with silica gel column to obtain the product. Silica gel column purification to obtain 0.15g of white solid with the yield of 36 percent. The product was characterized as follows:

¹H NMR(400MHz,CDCl₃):δ8.10(s,1H),7.65-7.97(m,4H),7.30-7.50(m,4H),7.17(m,2H),6.97(m,1H),6.21-6.25(m,2H),3.85(s,2H).

HR-MS(ESI):[M+H]+C₂₆H₁₈ClN₄O₂calculated 453.1118, found 453.1134.

Example 2

This example prepares compound 2: 8- (4- (2- (3-chlorobenzyl) -1H-indol-7-yl) -1H-1,2, 3-triazole-1-yl) -2H-chromen-2-one, the structure of which is shown as follows:

referring to the preparation method in example 1, 1-chloro-4- (iodomethyl) benzene was replaced with 1-chloro-3- (iodomethyl) benzene to obtain 0.17g of a white solid in a yield of 41%. The product was characterized as follows:

¹H NMR(400MHz,CDCl₃):δ8.09(s,1H),7.65-7.97(m,5H),7.30-7.50(m,3H),7.17-7.20(m,3H),6.97(m,1H),6.21-6.25(m,2H),3.85(s,2H).

HR-MS(ESI):[M+H]+C₂₆H₁₈ClN₄O₂calculated 453.1118, found 453.1145.

Example 3

This example prepares compound 3: 8- (4- (2- (2-chlorobenzyl) -1H-indol-7-yl) -1H-1,2, 3-triazole-1-yl) -2H-chromen-2-one, the structure of which is shown as follows:

referring to the preparation method in example 1, 1-chloro-4- (iodomethyl) benzene was replaced with 1-chloro-2- (iodomethyl) benzene to obtain 0.15g of a white solid in a yield of 38%. The product was characterized as follows:

¹H NMR(400MHz,CDCl₃):δ8.09(s,1H),7.65-7.97(m,4H),7.27-7.50(m,5H),6.97-7.11(m,2H),6.21-6.25(m,2H),3.85(s,2H).

HR-MS(ESI):[M+H]+C₂₆H₁₈ClN₄O₂calculated 453.1118, found 453.1139.

Example 4

This example prepares compound 4: 8- (4- (2- (4-methylbenzyl) -1H-indol-7-yl) -1H-1,2, 3-triazol-1-yl) -2H-chromen-2-one, the structure of which is shown below:

referring to the preparation method in example 1, 1-chloro-4- (iodomethyl) benzene was replaced with 1-methyl-4- (iodomethyl) benzene to obtain 0.20g of a white solid in 52% yield. The product was characterized as follows:

¹H NMR(400MHz,CDCl₃):δ8.07-8.12(m,3H),7.50-7.97(m,7H),7.42(m,1H),6.95(m,1H),6.21-6.25(m,2H),3.85(s,2H).

HR-MS(ESI):[M+H]+C₂₇H₂₁N₄O₂calculated 433.1665, found 433.1698.

Example 5

This example prepares compound 5: 8- (4- (2- (3-nitrobenzyl) -1H-indol-7-yl) -1H-1,2, 3-triazol-1-yl) -2H-chromen-2-one, the structure of which is shown below:

referring to the preparation method in example 1, 1-chloro-4- (iodomethyl) benzene was replaced with 1-nitro-3- (iodomethyl) benzene to obtain 0.15g of a white solid in a yield of 36%. The product was characterized as follows:

¹H NMR(400MHz,CDCl₃):δ8.08(s,1H),7.65-7.97(m,4H),7.42(m,1H),7.51(m,1H),7.11(m,4H),6.95(m,1H),6.21-6.25(m,2H),3.85(s,2H),2.35(s,3H).

HR-MS(ESI):[M+H]+C₂₆H₁₈N₅O₄calculated 464.1359, found 464.1378.

Example 6

This example prepares compound 6: 8- (4- (2- (2-cyanobenzyl) -1H-indol-7-yl) -1H-1,2, 3-triazol-1-yl) -2H-chromen-2-one, the structure of which is shown below:

referring to the preparation method in example 1, 1-chloro-4- (iodomethyl) benzene was replaced with 1-cyano-2- (iodomethyl) benzene to obtain 0.17g of a white solid in a yield of 42%. The product was characterized as follows:

¹H NMR(400MHz,CDCl₃):δ8.08(s,1H),7.61-7.97(m,5H),7.42-7.54(m,5H),6.97(m,1H),6.21-6.25(m,2H),3.85(s,2H).

HR-MS(ESI):[M+H]+C₂₇H₁₈N₅O₂calculated 444.1460, found 444.1493.

Example 7

This example prepares compound 7: 8- (4- (2- (4-fluorobenzyl) -1H-indol-7-yl) -1H-1,2, 3-triazole-1-yl) -2H-chromen-2-one, the structure of which is shown as follows:

referring to the preparation method in example 1, 1-chloro-4- (iodomethyl) benzene was replaced with 1-fluoro-4- (iodomethyl) benzene to obtain 0.19g of a white solid in 48% yield. The product was characterized as follows:

¹H NMR(400MHz,CDCl₃):δ8.10(s,1H),7.65-7.97(m,4H),7.50(m,1H),7.43(m,1H),6.97(m,1H),6.21-6.25(m,2H),3.85(s,2H).

HR-MS(ESI):[M+H]+C₂₆H₁₈FN₄O₂calculated 437.1414, found 437.1447.

Example 8

This example prepares compound 8: 8- (4- (2- (4-methanesulfonylbenzyl) -1H-indol-7-yl) -1H-1,2, 3-triazol-1-yl) -2H-chromen-2-one, which has the following structure:

referring to the preparation method in example 1, 1-chloro-4- (iodomethyl) benzene was replaced with 1-methanesulfonyl-4- (iodomethyl) benzene to obtain 0.19g of a white solid in a yield of 42%. The product was characterized as follows:

¹H NMR(400MHz,CDCl₃):δ8.08(s,1H),7.65-7.97(m,8H),7.50(m,1H),7.43(m,1H),6.97(m,1H),6.21-6.25(m,2H),3.85(s,2H),3.32(s,3H).

HR-MS(ESI):[M+H]+C₂₇H₂₁N₄O₄s calculated 497.1284, found 497.1299.

Example 9

This example prepares compound 9: 8- (4- (2- (4-trifluoromethyl benzyl) -1H-indol-7-yl) -1H-1,2, 3-triazole-1-yl) -2H-chromen-2-one, the structure of which is shown as follows:

referring to the preparation method in example 1, 1-chloro-4- (iodomethyl) benzene was replaced with 1-trifluoromethyl-4- (iodomethyl) benzene to obtain 0.21g of a white solid in a yield of 47%. The product was characterized as follows:

¹H NMR(400MHz,CDCl₃):δ8.08(s,1H),7.65-7.97(m,4H),7.42-7.50(m,4H),7.16(m,2H),6.97(m,1H),6.21-6.25(m,2H),3.85(s,2H).

HR-MS(ESI):[M+H]+C₂₇H₂₁N₄O₄s calculated 497.1284, found 497.1299.

Example 10

This example prepares compound 10: 8- (4- (2- (4-phenylbenzyl) -1H-indol-7-yl) -1H-1,2, 3-triazol-1-yl) -2H-chromen-2-one, which has the following structure:

referring to the preparation method in example 1, 1-chloro-4- (iodomethyl) benzene was replaced with 1-phenyl-4- (iodomethyl) benzene to obtain 0.14g of a white solid in a yield of 31%. The product was characterized as follows:

¹H NMR(400MHz,CDCl₃):δ8.08(s,1H),7.65-7.97(m,4H),7.41-7.50(m,7H),7.29-7.33(m,4H),6.97(m,1H),6.21-6.25(m,2H),3.85(s,2H).

HR-MS(ESI):[M+H]+C₃₂H₂₃N₄O₂calculated 495.1821, found 495.1845.

Example 11

This example prepares compound 11: 8- (4- (2- (4-hydroxybenzyl) -1H-indol-7-yl) -1H-1,2, 3-triazole-1-yl) -2H-chromen-2-one, the structure of which is shown as follows:

referring to the preparation method in example 1, 1-chloro-4- (iodomethyl) benzene was replaced with 1-hydroxy-4- (iodomethyl) benzene to obtain 0.14g of a white solid in a yield of 35%. The product was characterized as follows:

¹H NMR(400MHz,CDCl₃):δ8.08(s,1H),7.65-7.97(m,4H),7.50(m,1H),7.42(m,1H),7.06(m,2H),6.97(m,1H),6.65(m,2H),6.21-6.25(m,2H),3.85(s,2H).

HR-MS(ESI):[M+H]+C₂₆H₁₉N₄O₃calculated 435.1457, found 435.1487.

Example 12

This example prepares compound 12: 8- (4- (2- (4-methoxybenzyl) -1H-indol-7-yl) -1H-1,2, 3-triazol-1-yl) -2H-chromen-2-one, the structure of which is shown below:

referring to the preparation method in example 1, 1-chloro-4- (iodomethyl) benzene was replaced with 1-methoxy-4- (iodomethyl) benzene to obtain 0.19g of a white solid in a yield of 47%. The product was characterized as follows:

¹H NMR(400MHz,CDCl₃):δ8.08(s,1H),7.65-7.97(m,4H),7.50(m,1H),7.42(m,1H),7.12(m,2H),6.97(m,1H),6.87(m,2H),6.21-6.25(m,2H),3.85(s,2H).

HR-MS(ESI):[M+H]+C₂₇H₂₁N₄O₃calculated 449.1614, found 449.1654.

Example 13

This example prepares compound 13: 8- (4- (2- (3, 4-difluorobenzyl) -1H-indol-7-yl) -1H-1,2, 3-triazol-1-yl) -2H-chromen-2-one, the structure of which is shown below:

referring to the preparation method in example 1, 1-chloro-4- (iodomethyl) benzene was replaced with 1, 2-fluoro-4- (iodomethyl) benzene to obtain 0.25g of a white solid in a yield of 61%. The product was characterized as follows:

¹H NMR(400MHz,CDCl₃):δ8.08(s,1H),7.65-7.97(m,4H),7.50(m,1H),7.42(m,1H),6.75-7.10(m,4H),6.21-6.25(m,2H),3.85(s,2H).

HR-MS(ESI):[M+H]+C₂₆H₁₇F₂N₄O₂calculated 455.1320, found 455.1337.

Example 14

This example prepares compound 14: 8- (4- (2- (3-bromo-4-chlorobenzyl) -1H-indol-7-yl) -1H-1,2, 3-triazol-1-yl) -2H-chromen-2-one, the structure of which is shown below:

referring to the preparation method in example 1, 1-chloro-4- (iodomethyl) benzene was replaced with 1-chloro-2-bromo-4- (iodomethyl) benzene to obtain 0.16g of a white solid in a yield of 35%. The product was characterized as follows:

¹H NMR(400MHz,CDCl₃):δ8.08(s,1H),7.65-7.97(m,4H),7.50(m,1H),7.42(m,1H),7.32(m,1H),7.26(m,1H),7.11(d,J＝5.4Hz,1H),6.97(m,1H)6.21-6.25(m,2H),3.85(s,2H).

HR-MS(ESI):[M+H]+C₂₆H₁₇BrClN₄O₂calculated 531.0223, found 531.0257.

Example 15

This example prepares compound 15: 8- (4- (2- (7-bromonaphthalen-1-yl) methyl) -1H-indol-7-yl) -1H-1,2, 3-triazol-1-yl) -2H-chromen-2-one, the structure of which is shown below:

referring to the preparation method in example 1, 1-chloro-4- (iodomethyl) benzene was replaced with 2-bromo-8- (iodomethyl) naphthalene to obtain 0.22g of a white solid in a yield of 43%. The product was characterized as follows:

¹H NMR(400MHz,CDCl₃):δ8.29(s,1H),8.08(s,1H),7.65-7.97(m,7H),7.47-7.50(m,3H),7.07(m,1H),6.97(m,1H)6.21-6.25(m,2H),4.29(s,2H).

HR-MS(ESI):[M+H]+C₃₀H₂₀BrN₄O₂calculated 547.0770, found 547.0797.

Example 16

This example prepares compound 16: 8- (4- (2- (3-chloronaphthalene-1-yl) methyl) -1H-indol-7-yl) -1H-1,2, 3-triazole-1-yl) -2H-chromen-2-one, the structure of which is shown as follows:

referring to the preparation method in example 1, 1-chloro-4- (iodomethyl) benzene was replaced with 2-chloro-4- (iodomethyl) naphthalene to obtain 0.23g of a white solid in 52% yield. The product was characterized as follows:

¹H NMR(400MHz,CDCl₃):δ8.18(s,1H),8.08-8.05(m,2H),7.65-7.97(m,4H),7.42-7.54(m,5H),7.11(m,1H),6.97(m,1H)6.21-6.25(m,2H),4.29(s,2H).

HR-MS(ESI):[M+H]+C₃₀H₂₀ClN₄O₂calculated 503.1275, found 503.1298.

Example 17

This example prepares compound 17: 8- (4- (2- (quinolin-5-yl) methyl) -1H-indol-7-yl) -1H-1,2, 3-triazol-1-yl) -2H-chromen-2-one, which has the following structure:

referring to the preparation method in example 1, 1-chloro-4- (iodomethyl) benzene was replaced with 5- (iodomethyl) quinoline to obtain 0.16g of a white solid in a yield of 38%. The product was characterized as follows:

¹H NMR(400MHz,CDCl₃):δ8.80(m,1H),8.48(m,1H),8.05(s,1H),7.65-7.97(m,6H),7.42-7.54(m,4H),6.97(m,1H)6.21-6.25(m,2H),3.85(s,2H).

HR-MS(ESI):[M+H]+C₂₉H₂₀N₅O₂calculated 470.1617, found 470.1653.

Example 18

This example prepares compound 18: 8- (4- (2- (7-bromo-quinolin-5-yl) methyl) -1H-indol-7-yl) -1H-1,2, 3-triazol-1-yl) -2H-chromen-2-one, which has the following structure:

referring to the preparation method in example 1, 1-chloro-4- (iodomethyl) benzene was replaced with 3-bromo-5- (iodomethyl) quinoline to give 0.18g of a white solid in a yield of 38%. The product was characterized as follows:

¹H NMR(400MHz,CDCl₃):δ8.92(s,1H),8.78(s,1H),8.08(s,1H),7.65-7.97(m,6H),7.42-7.56(m,3H),6.97(m,1H)6.21-6.25(m,2H),3.85(s,2H).

HR-MS(ESI):[M+H]+C₂₉H₁₉BrN₅O₂calculated 548.0722, found 548.0756.

Test example 1

In vitro activity screening of SIRT2 inhibitors:

the full-length expression sequence ORF of the human SIRT2 gene is 1170bp (Access No.: NM-012237), and the size of the expressed SIRT2 protein is 43 KDa. The expression and purification steps of the SIRT2 recombinant protein are as follows: the SIRT2 gene was cloned into an expression vector pET-15b (primers as follows Forward: 5'-TAATACGACTCACTATAGGG-3'; backward: 5'-TTCACTTCTGAGTTCGGCATG-3'), and the expressed SIRT2 protein contained the tag His at the N-terminus for purification₆. The plasmid was transformed into E.coli BL2(DE3), induced with 1mM IPTG and expressed at 18 ℃ for 6 hours. The induced E.coli cells were collected by centrifugation and frozen at-20 ℃. The bacterial pellet was resuspended in 15mL of cell lysate (50mM Tris-HCl pH 8.0,300mM NaCl) and the bacterial sample was sonicated for 10min before centrifugation (12000 Xg, 20min) at 4 ℃ to remove the pellet. The recombinant expressed soluble SIRT2 protein was present in the supernatant. Passing the supernatant through Ni²⁺Purification with NTA-agarose matrix column (Qiagen). During the purification process, the non-bound protein was washed off with the loading solution (50mM Tris-HCl pH 8.0,300mM NaCl) first, and then the non-specifically bound protein was washed off by gradually increasing the concentration of imidazole (0-200mM), resulting in purified recombinant SIRT2 protein. Imidazole was removed using a PD-10 column and the purified SIRT2 protein concentration was determined using the Bradford method. The recombinant SIRT2 protein was stored in storage medium (50mM Tris-HCl, pH 8.0,265mM NaCl,0.2mM DTT, and 10% glycerol) and placed at-20 ℃.

According to the report in the literature, a SIRT2 enzyme activity measuring method is established. Firstly synthesizing a substrate peptide segment Ac-Gln-Pro-Lys- [ Lys- (Ac) of SIRT2]AMC as a substrate in the enzyme activity assay. The attached fluorescent tag is AMC (7-Amino-4-methylicuomarin). The whole determination process comprises two steps: the catalytic reaction was carried out in 60. mu.L of a reaction solution (25mM Tris-HCl, pH 8.0,137mM NaCl,2.7mM KCl,1mM MgCl)₂and 1mg/ml BSA), 500. mu.M NAD was added⁺50 μ M substrate peptide fragment, 1.0 μ g SIRT2 and varying concentrations of compounds 1-18. The reaction was left at 37 ℃ for 2 hours. In this reaction, the acetyl group of the Lysine residue on the small molecule peptide fragment is removed to varying degrees. Then, 60. mu.L of the sample treatment solution (50mM Tris-HCl, pH 8.0,100mM NaCl, Trypsin and 4mM nicotinamide) was added to the reaction solution and mixed, and left at 37 ℃ for 20 minutes. The microplate reader is set to have excitation light of 355nm and absorption light of 460nm, and the absorption intensity is measured. There were 2 replicates per compound concentration in the assay, with appropriate controls, and the results of the assay were calculated using the processing software GraphPad Prism for the inhibitory activity IC of each inhibitor₅₀. The results are shown in table 1 (where AGK is the positive control):

TABLE 1

As can be seen from the data in Table 1: 18 tested samplesIn vitro SIRT2 inhibitory Activity of Compounds IC₅₀All reach micromolar level, especially IC of 13 compounds₅₀Value reaches 10^-7mol/L level, wherein IC of 3 compounds₅₀Value reaches 10^{- 8}The mol/L level shows that the 8- (indole triazole) substituted coumarin compound has good SIRT2 inhibitory activity.

Test example 2

Cytotoxicity assay of compounds against neuroma cells SH-SY 5Y:

this test example measured the cytotoxicity of the compound against SH-SY5Y neuroma cells and was used to determine the dose for the next Parkinson cell model protection test. For the determination of cytotoxicity, the specific experimental procedure was as follows: cultured neuroma cells SH-SY5Y were seeded in 96-well cell culture plates at about 6000 cells per well. After overnight incubation, the culture medium was replaced with fresh medium and different concentrations of compounds 1-18(0.01,0.05,0.1,0.5,1,5,10,20,50,100, 200. mu.M) were added to each well and different control samples were set. The cells were cultured at 37 ℃ for another 48 hours. Add 10. mu.L of dye to each well

(Invitrogen) the incubation was continued at 37 ℃ for about 2 hours, and the color change of the staining solution was observed, and the change in absorbance was read under a microplate reader (Ex: 530 nm; Em: 590 nm). There were 2 replicates for each concentration of compound tested. Experimental results cytotoxicity CC of each compound was calculated using data processing software GraphPad Prism₅₀. The results are shown in table 2 (where Taxol is the positive control group):

TABLE 2

As can be seen from the data in Table 2: the tested 18 compounds have cytotoxicity CC50 of more than 20 mu M to SH-SY5Y, namely the 8- (indole triazole) substituted coumarin compound has no obvious inhibitory activity to SH-SY5Y cells.

Test example 3

Screening compounds with protective activity against parkinson's disease cell models:

for screening test of protective effect of Parkinson's disease cell model, currently generally accepted nerve agent MPP is adopted⁺(5mM) acts on a neuroma cell SH-SY5Y to construct a Parkinson disease cell model. This nerve agent acts on SH-SY5Y cells to affect abnormal accumulation of alpha-synuclein in the cells and causes neurotoxicity similar to Parkinson's disease. The specific experimental method is as follows: approximately 20000 SH-SY5Y cells were seeded in 96-well cell plates and cultured overnight. Control test control with no nerve agent, and nerve agent MPP in each well of other groups⁺(5mM), MPP + group as no protective compound, compounds 1-18 (10. mu.M), respectively, were added simultaneously to the wells of the protection assay (where AGK is the positive control), and incubation was continued for 48 hours to measure the cell activity of the Parkinson's disease cell model and thereby evaluate their protective effect on the Parkinson's cell model. GraphPad Prism was used for data analysis of the test results. The results are shown in FIG. 1.

As can be seen from fig. 1: 18 compounds tested all were able to protect MPP to some extent⁺The damage of the nerve toxin to SH-SY5Y cells shows that the 8- (indole triazole) substituted coumarin compound has a protective effect on neuroma cells; and the compound 7, 9, 13, 14 has more obvious protective effect on neuroma cells.

The applicant states that the invention is illustrated by the above examples, but the invention is not limited to the above examples, that is, the invention is not limited to the above examples. It should be understood by those skilled in the art that any modification of the present invention, equivalent substitutions of the raw materials of the product of the present invention, addition of auxiliary components, selection of specific modes, etc., are within the scope and disclosure of the present invention.

The preferred embodiments of the present invention have been described in detail, however, the present invention is not limited to the specific details of the above embodiments, and various simple modifications may be made to the technical solution of the present invention within the technical idea of the present invention, and these simple modifications are within the protective scope of the present invention.

It should be noted that the various technical features described in the above embodiments can be combined in any suitable manner without contradiction, and the invention is not described in any way for the possible combinations in order to avoid unnecessary repetition.

Claims (10)

1. The 8- (indole triazole) substituted coumarin compound is characterized in that the structure of the 8- (indole triazole) substituted coumarin compound is shown as the formula (I):

wherein ring A is independently selected from phenyl, naphthyl, 5-14 membered aromatic heterocyclic group;

said phenyl, naphthyl, 5-14 membered aromatic heterocyclic group being unsubstituted or substituted by 1-5R_aSubstitution; r_aEach independently selected from deuterium, halogen, hydroxy, mercapto, amino, cyano, nitro, azido, methanesulfonyl, isopropylsulfonyl, benzenesulfonyl, aminosulfonyl, methanesulfonate, isopropylsulfonate, benzenesulfonate, trifluoromethyl, trifluoromethyloxy, carboxamido, C_1-8Alkyloxy, C_1-8Alkylcarbonyl group, C_1-8Alkyl oxycarbonyl radical, C_1-8Alkyl radical, C_3-8Cycloalkyl, phenyl;

in said R_aIn (A), the C_1-8Alkyl radical, C_3-8Cycloalkyl, phenyl unsubstituted or substituted by 1-5R_bSubstitution; the R is_bEach independently selected from deuterium, halogenPlain, hydroxy, mercapto, amino, cyano, nitro, azido, methylsulfonyl, isopropylsulfonyl, phenylsulfonyl, aminosulfonyl, methanesulfonate, isopropylsulfonate, benzenesulfonate, trifluoromethyl, trifluoromethyloxy, carboxamido, C_1-8An alkylcarbonyl group.

2. The 8- (indoletriazole) substituted coumarin compound of claim 1, wherein in formula (I), ring a is independently selected from phenyl, naphthyl, 5-14 membered aromatic heterocyclic group;

said phenyl, naphthyl, 5-14 membered aromatic heterocyclic group being unsubstituted or substituted by 1-5R_aSubstitution; r_aEach independently selected from deuterium, halogen, hydroxy, mercapto, amino, cyano, nitro, azido, methanesulfonyl, isopropylsulfonyl, benzenesulfonyl, aminosulfonyl, methanesulfonate, isopropylsulfonate, benzenesulfonate, trifluoromethyl, trifluoromethyloxy, carboxamido, C_1-6Alkyloxy, C_1-6Alkylcarbonyl group, C_1-6Alkyl oxycarbonyl radical, C_1-6Alkyl radical, C_3-6Cycloalkyl, phenyl;

in said R_aIn (A), the C_1-6Alkyl radical, C_3-6Cycloalkyl, phenyl unsubstituted or substituted by 1-5R_bSubstitution; the R is_bEach independently selected from deuterium, halogen, hydroxy, mercapto, amino, cyano, nitro, azido, methanesulfonyl, isopropylsulfonyl, benzenesulfonyl, aminosulfonyl, methanesulfonate, isopropylsulfonate, benzenesulfonate, trifluoromethyl, trifluoromethyloxy, carboxamido, C_1-6An alkylcarbonyl group.

3. The 8- (indoletriazole) substituted coumarin compound as claimed in claim 1 or 2, wherein the structure of the 8- (indoletriazole) substituted coumarin compound is shown as formula (IA):

wherein R is₁、R₂、R₃、R₄、R₅Independently selected from hydrogen, deuterium, halogen, hydroxy, mercapto, amino, cyano, nitro, azido, methanesulfonyl, isopropylsulfonyl, benzenesulfonyl, aminosulfonyl, methanesulfonate, isopropylsulfonate, benzenesulfonate, trifluoromethyl, trifluoromethyloxy, carboxamido, C_1-4Alkyloxy, C_1-4Alkylcarbonyl group, C_1-4Alkyl oxycarbonyl radical, C_1-4Alkyl, phenyl.

4. The 8- (indoletriazole) substituted coumarin compound as claimed in claim 1 or 2, wherein the structure of the 8- (indoletriazole) substituted coumarin compound is as shown in formula (IB):

wherein R is₁、R₂、R₃、R₄、R₅Independently selected from hydrogen, deuterium, halogen, hydroxy, mercapto, amino, cyano, nitro, methylsulfonyl, trifluoromethyl, trifluoromethyloxy, carboxamido, C_1-4Alkyloxy, C_1-4Alkylcarbonyl group, C_1-4An alkyl group; x is independently selected from N or C.

5. The 8- (indoletriazole) substituted coumarin compound as claimed in claim 1 or 2, wherein the 8- (indoletriazole) substituted coumarin compound is selected from the following structures:

6. the stereoisomer of the 8- (indoletriazole) substituted coumarin compound, the pharmaceutically acceptable salt thereof or the pharmaceutical composition containing the stereoisomer of the 8- (indoletriazole) substituted coumarin compound according to any one of claims 1-5;

preferably, the pharmaceutical composition further comprises pharmaceutically acceptable pharmaceutical excipients.

7. The preparation method of the 8- (indoletriazole) substituted coumarin compound as claimed in any one of claims 1-5, characterized in that the preparation method comprises the following steps:

(1) will be provided with

Mixing with 7- (1- (1- (2-oxo) -2H-chromen-8-yl) -1H-1,2, 3-triazol-4-yl) -1H-indole-1-carboxylic acid tert-butyl ester, and reacting to obtain an intermediate product, namely 2- (4-chlorobenzyl) -7- (1- (2-oxo-2H-chromen-8-yl) -1H-1,2, 3-triazol-4-yl) -1H-indole-1-carboxylic acid tert-butyl ester;

(2) hydrolyzing 2- (4-chlorobenzyl) -7- (1- (2-oxo-2H-chromen-8-yl) -1H-1,2, 3-triazole-4-yl) -1H-indole-1-tert-butyl formate under acidic condition, and adjusting pH to neutrality to obtain the final product;

wherein the defined range of ring a is in accordance with the range defined in any one of claims 1 to 5;

the reaction formula is shown as follows:

8. the preparation method of the 8- (indoletriazole) substituted coumarin compound as claimed in claim 7, wherein the preparation method of the 7- (1- (1- (2-oxo) -2H-chromen-8-yl) -1H-1,2, 3-triazol-4-yl) -1H-indole-1-carboxylic acid tert-butyl ester in step (1) comprises the following steps:

(1) reacting 8-bromocoumarin with NaN₃Mixing and reacting to obtain 8-azido coumarin;

(2) mixing 8-azido coumarin with 7-ethynyl-1H-indole and cuprous iodide, and reacting to obtain 8- (4- (1H-indol-7-yl) -1H-1,2, 3-triazol-1-yl) -2H-chromen-2-one;

(3) mixing 8- (4- (1H-indol-7-yl) -1H-1,2, 3-triazol-1-yl) -2H-chromen-2-one with sodium hydride, and adding thereto (Boc)₂O to give 7- (1- (1- (2-oxo) -2H-chromen-8-yl) -1H-1,2, 3-triazol-4-yl) -1H-indole-1-carboxylic acid tert-butyl ester;

the reaction formula is shown as follows:

9. the 8- (indoletriazole) substituted coumarin compound according to any one of claims 1-5 or the 8- (indoletriazole) substituted coumarin compound according to claim 6, and the pharmaceutically acceptable salt and the pharmaceutical composition containing the stereoisomer are applied to preparation of drugs for treating and/or preventing diseases or symptoms related to SIRT2 overhigh activity or SIRT2 overexpression.

10. The use of claim 9, wherein the disease or condition associated with excessive SIRT2 activity or overexpression of SIRT2 comprises parkinson's disease, metabolic disease, or tumor.

Images