Compounds having sirt6 agonistic activity and use thereof

Abstract

A class of compounds represented by formula (I) or a stereoisomer, a tautomer, a solvate, or a pharmaceutically acceptable salt thereof.

Description

Compounds with SIRT6 agonist activity and uses thereof

[0001] Priority information

[0002] This application claims priority and benefits of patent application 202311679025.4 filed with the State Intellectual Property Office of China on December 8, 2023, and incorporates the entire text of it herein by reference. Technical Field

[0003] The present invention relates to the field of biomedicine, and more specifically, to compounds having SIRT6 agonist activity and uses thereof. Background Art

[0004] SIRT6, as an important member of the III family (Sirtuin1-7) of histone deacetylases (HDACs), was first reported in 2000 and has been widely studied in recent years. With the deepening of research, many physiological functions of SIRT6 have been discovered. In addition to its NAD + In addition to its histone deacetylase activity, it also has mono ADP-ribosyltransferase and de-fatty acylation functions.

[0005] Studies have shown that SIRT6 activity is regulated by multiple factors, primarily by catalyzing the deacetylation of related proteins within the cell. It is involved in vital activities such as DNA repair, inflammation, and glucose and lipid metabolism, and is a key drug target for aging-related diseases, inflammation, cancer, and other diseases. Therefore, the development of drugs that activate SIRT6 deacetylase activity is of great significance for delaying aging, treating aging-related diseases, and achieving healthy aging.

[0006] Although several agonists targeting SIRT6 deacetylase activity, such as MDL-800 and MDL-811, have been shown to have anti-inflammatory effects, screening for SIRT6 agonists with lower toxicity and improved efficacy remains of great scientific value. Therefore, the development of drugs with enhanced SIRT6 agonist activity is urgently needed. Summary of the Invention

[0007] The present invention aims to solve, at least to some extent, at least one of the technical problems existing in the prior art. To this end, the present invention provides a class of compounds having SIRT6 agonist activity and uses thereof. Such compounds have SIRT6 agonist activity and can be used as SIRT6 agonists to effectively prevent and / or treat diseases mediated by the SIRT6 protein.

[0008] In the first aspect of the present invention, the present invention provides a class of compounds, which are compounds represented by formula (I) or stereoisomers, tautomers, solvates, and pharmaceutically acceptable salts thereof:

[0009] wherein ring A is selected from a polyvalent aromatic ring; R is selected from H, halogen, hydroxyl, amino, carbonyl, C optionally substituted by R1 1～6 Alkyl, optionally substituted by R1C 1～6 Alkoxy, optionally substituted by R1 C 1～6 Haloalkyl or optionally substituted by R1 C 1～6 Halogenated oxyalkyl; R1 is selected from C 1～4 Alkyl, hydroxy, amino, nitro, cyano, oxo, carboxyl, carbonyl.

[0010] The compounds of the present invention have SIRT6 agonist activity and can be used as SIRT6 agonists to effectively prevent and / or treat diseases mediated by SIRT6 protein.

[0011] According to an embodiment of the present invention, ring A is selected from 5- to 10-membered aromatic rings.

[0012] According to an embodiment of the present invention, ring A is selected from a six-membered aromatic ring.

[0013] According to an embodiment of the present invention, R is selected from H, halogen or C 1～6 Alkoxy.

[0014] According to an embodiment of the present invention, ring A is selected from a benzene ring.

[0015] According to an embodiment of the present invention, R is selected from C 1～6 Alkoxy.

[0016] According to an embodiment of the present invention, the compound represented by formula (I) has a structure represented by the following formula (II):

[0017] According to an embodiment of the present invention, the compound represented by formula (I) has the structure shown below:

[0018] In a second aspect of the present invention, a pharmaceutical composition is provided. According to an embodiment of the present invention, the pharmaceutical composition comprises: the compound described in the first aspect, or a stereoisomer, tautomer, solvate, or pharmaceutically acceptable salt thereof. As mentioned above, the compound described in the first aspect has SIRT6 agonist activity. Therefore, the pharmaceutical composition of the present invention can activate and / or promote the expression or activity of SIRT6 protein, thereby effectively preventing and / or treating diseases mediated by SIRT6 protein.

[0019] According to an embodiment of the present invention, the pharmaceutical composition further includes a pharmaceutically acceptable carrier or excipient.

[0020] In a third aspect, the present invention provides the use of the compound described in the first aspect as a SIRT6 agonist. As previously mentioned, the compound described in the first aspect has SIRT6 agonist activity and, as a SIRT6 agonist, can activate and / or promote the expression or activity of SIRT6 protein in cells, or effectively prevent and / or treat diseases mediated by SIRT6 protein.

[0021] In a fourth aspect, the present invention provides the use of the compound described in the first aspect in preparing a reagent for activating and / or promoting the expression or activity of SIRT6 protein. As previously mentioned, the compound described in the first aspect has SIRT6 agonist activity and can be used as a reagent to effectively activate and / or promote the expression or activity of SIRT6 protein. It is particularly useful for increasing the expression or activity of SIRT6 protein in cells in vitro, for example, to construct a desired cell model.

[0022] In a fifth aspect, the present invention provides the use of the compound described in the first aspect or the pharmaceutical composition described in the second aspect in the preparation of a medicament for preventing and / or treating diseases or symptoms mediated by the SIRT6 protein. As previously noted, the compound described in the first aspect exhibits SIRT6 agonist activity and, as a SIRT6 medicament, can effectively prevent and / or treat diseases or symptoms mediated by the SIRT6 protein.

[0023] In a sixth aspect, the present invention provides the use of the compound described in the first aspect or the pharmaceutical composition described in the second aspect for preventing and / or treating diseases or symptoms mediated by the SIRT6 protein. As previously noted, the compound described in the first aspect exhibits SIRT6 agonist activity and can effectively prevent and / or treat diseases or symptoms mediated by the SIRT6 protein.

[0024] According to an embodiment of the present invention, the related diseases or symptoms mediated by the SIRT6 protein include but are not limited to aging-related diseases, cancer, inflammation, and anti-aging.

[0025] According to an embodiment of the present invention, the aging-related diseases include but are not limited to osteoarthritis, intervertebral disc herniation, osteoporosis, Alzheimer's disease, atherosclerosis, liver fibrosis, age-related macular degeneration, diabetic nephropathy, and idiopathic pulmonary fibrosis.

[0026] It should be noted that the cancer described in the present invention can inhibit its development or growth by activating or promoting the expression of SIRT6 protein. According to an embodiment of the present invention, the cancer is selected from but not limited to bladder cancer, nasopharyngeal carcinoma or glioblastoma.

[0027] In a seventh aspect, the present invention provides a method for activating and / or promoting SIRT6 protein activity in cells in vitro. According to an embodiment of the present invention, the method comprises: contacting the cells with the compound described in the first aspect. As previously mentioned, the compound described in the first aspect has SIRT6 agonist activity. Thus, contacting the above-mentioned compound with cells can activate and / or promote the expression or activity of SIRT6 protein in the cells.

[0028] In an eighth aspect, the present invention provides a method for preventing and / or treating diseases or symptoms mediated by the SIRT6 protein. According to an embodiment of the present invention, the method comprises administering a pharmaceutically acceptable amount of the compound described in the first aspect or the pharmaceutical composition described in the second aspect to a subject. As previously mentioned, the compound described in the first aspect has SIRT6 agonist activity. Thus, the use of the above-mentioned compound can effectively prevent and / or treat diseases or symptoms mediated by the SIRT6 protein.

[0029] Additional aspects and advantages of the present invention will be set forth in part in the description which follows and, in part, will be obvious from the description which follows, or may be learned by practice of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] The above and / or additional aspects and advantages of the present invention will become apparent and readily understood from the following description of the embodiments with reference to the accompanying drawings, in which:

[0031] FIG1 is a hydrogen spectrum of compound 1 in Example 1;

[0032] FIG2 is the detection result of SIRT6 enzyme activity of compound 1 in Example 2;

[0033] FIG3 is the test result of the dose-effect curve of compound 1 in Example 2;

[0034] FIG4 shows the toxicity test results of Compound 1 using CCK-8 in Example 2;

[0035] FIG5 shows the results of Western blotting to detect the activation effect of compound 1 on SIRT6 protein activity in cell lines in Example 2. DETAILED DESCRIPTION

[0036] The embodiments of the present invention are described in detail below. The embodiments described below are exemplary and are only used to explain the present invention, and should not be understood as limiting the present invention.

[0037] It should be noted that the terms "first" and "second" are used for descriptive purposes only and should not be understood to indicate or imply relative importance or implicitly specify the number of the technical features indicated. Therefore, features defined as "first" or "second" may explicitly or implicitly include one or more of such features. Furthermore, in the description of the present invention, unless otherwise specified, "plurality" means two or more.

[0038] Definitions and General Terms

[0039] In this document, the terms “include” or “comprising” are open expressions, that is, including the contents specified in the present invention, but not excluding other contents.

[0040] As used herein, the terms "optionally," "optional," or "optionally" generally mean that the subsequently described event or circumstance may but need not occur, and that the description includes instances where the event or circumstance occurs and instances where it does not.

[0041] As used herein, the definitions and conventions of stereoisomers are generally in accordance with SP Parker, Ed., McGraw-Hill Dictionary of Chemical Terms (1984) McGraw-Hill Book Company, New York; and Eliel, E. and Wilen, S., "Stereochemistry of Organic Compounds", John Wiley & Sons, Inc., New York, 1994. The compounds of the present invention may contain asymmetric centers or chiral centers and therefore exist in different stereoisomeric forms. It is contemplated that all stereoisomeric forms of the compounds of the present invention, including but not limited to diastereomers, enantiomers and atropisomers, and mixtures thereof, such as racemic mixtures, are encompassed by the present invention. Many organic compounds exist in optically active forms, i.e., they have the ability to rotate the plane of plane-polarized light. When describing an optically active compound, the prefixes D and L or R and S are used to denote the absolute configuration of the molecule with respect to the chiral center(s) in the molecule. The prefixes d and l, or (+) and (-), are symbols used to designate the rotation of plane-polarized light caused by a compound, where (-) or l indicates that the compound is levorotatory. A compound prefixed with (+) or d is dextrorotatory. With respect to a given chemical structure, these stereoisomers are identical except that they are mirror images of one another. Specific stereoisomers may also be referred to as enantiomers, and a mixture of such isomers is often referred to as a mixture of enantiomers. A 50:50 mixture of enantiomers is called a racemic mixture or racemate, which can occur when there is no stereoselectivity or stereospecificity in a chemical reaction or process.

[0042] Depending on the choice of starting materials and process, the compounds of the present invention may exist as one of the possible isomers or as a mixture thereof, for example as pure optical isomers, or as a mixture of isomers, such as a racemic and diastereomeric mixture, depending on the number of asymmetric carbon atoms. Optically active (R)- or (S)-isomers can be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques. If the compound contains a double bond, the substituents may be in the E or Z configuration; if the compound contains a disubstituted cycloalkyl group, the cycloalkyl substituents may be in the cis or trans configuration.

[0043] The compounds of the present invention may contain asymmetric centers or chiral centers and therefore exist in different stereoisomeric forms. It is contemplated that all stereoisomeric forms of the compounds of the present invention, including but not limited to diastereomers, enantiomers and atropisomers and geometric (or conformational) isomers and mixtures thereof, such as racemic mixtures, are within the scope of the present invention.

[0044] As used herein, the term "tautomer" or "tautomeric form" refers to structural isomers with different energies that can be converted into each other through a low energy barrier. If tautomerism is possible (such as in solution), a chemical equilibrium of the tautomers can be reached. For example, proton tautomers (also known as prototropic tautomers) include interconversions that occur through proton migration, such as keto-enol isomerization and imine-enamine isomerization. Valence tautomers include interconversions that occur through the reorganization of some bonding electrons. Unless otherwise indicated, all tautomeric forms of the compounds of the present invention are within the scope of the present invention.

[0045] As used herein, the term "solvate" refers to an association formed between one or more solvent molecules and a compound of the present invention. Solvents that form solvates include, but are not limited to, water, isopropanol, ethanol, methanol, dimethyl sulfoxide, ethyl acetate, acetic acid, and aminoethanol. The term "hydrate" refers to an association formed when the solvent molecule is water.

[0046] As used herein, the term "pharmaceutically acceptable" means that the substance or composition must be chemically and / or toxicologically compatible with the other ingredients of the formulation and / or the mammal to be treated therewith. Preferably, the term "pharmaceutically acceptable" as used herein means approved by federal regulatory agencies or national governments or listed in the U.S. Pharmacopoeia or other generally recognized pharmacopeia for use in animals, particularly humans.

[0047] As used herein, the term "pharmaceutically acceptable salts" refers to organic and inorganic salts of the compounds of the present invention. Pharmaceutically acceptable salts are well known in the art, as described in SM Berge et al., describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 1977, 66: 1-19.

[0048] As used herein, the term "halogen" refers to a fluorine, chlorine, bromine or iodine atom.

[0049] In this article, the term “C 1～6 "Alkyl" refers to a linear or branched saturated monovalent hydrocarbon group having 1, 2, 3, 4, 5 or 6 carbon atoms, such as C 1～5 Alkyl, C 1～4 Alkyl, C 1～3 Alkyl, C 2～5 Alkyl, C 2～4 Alkyl, C 2～3 Alkyl. These include, but are not limited to, methyl, ethyl, n-propyl (n-Pr, -CH2CH2CH3), isopropyl (i-Pr, -CH(CH3)2), n-butyl (n-Bu, -CH2CH2CH2CH3), isobutyl (i-Bu, -CH2CH(CH3)2), sec-butyl (s-Bu, -CH(CH3)CH2CH3), tert-butyl (t-Bu, -C(CH3)3), n-pentyl (-CH2CH2CH2C H2CH3), 2-pentyl (-CH(CH3)CH2CH2CH3), 3-pentyl (-CH(CH2CH3)2), 2-methyl-2-butyl (-C(CH3)2CH2CH3), 3-methyl-2-butyl (-CH(CH3)CH(CH3)2), 3-methyl-1-butyl (-CH2CH2CH(CH3)2), 2-methyl-1-butyl (-CH2CH(CH3)CH2CH3), n-hexyl ( -CH2CH2CH2CH2CH2CH3), 2-hexyl (-CH(CH3)CH2CH2CH2CH3), 3-hexyl (-CH(CH2CH3)(CH2CH2CH3)), 2-methyl-2-pentyl (-C(CH3)2CH2CH2CH3), 3-methyl-2-pentyl (-CH(CH3)CH(CH3)CH2CH3), 4-methyl-2-pentyl (-CH(CH3)CH2CH(CH3)2), 3-methyl-3-pentyl (-C(CH3)(CH2CH3)2), 2-methyl-3-pentyl (-CH(CH2CH3)CH(CH3)2), 2,3-dimethyl-2-butyl (-C(CH3)2CH(CH3)2), 3,3-dimethyl-2-butyl (-CH(CH3)C(CH3)3), wherein the alkyl groups may independently be unsubstituted or substituted with one or more substituents described herein.

[0050] As used herein, the term "haloalkyl" refers to an alkyl group in which one or more H atoms are replaced by any halogen.

[0051] In this article, the term “C 1～6 "Alkoxy" refers to a C 1～6Alkyl, wherein the term "alkyl" is as defined above. For example: methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, sec-butoxy, pentoxy, isopentoxy and n-hexoxy, or isomers of the above groups. In particular, the "C1-6 alkoxy" may contain 1, 2, 3, 4 or 5 carbon atoms ("C 1～5 Alkoxy”), preferably, may contain 1, 2, 3 or 4 carbon atoms (“C 1～4 alkoxy").

[0052] As used herein, the term "halooxyalkyl" refers to an oxyalkyl group in which one or more H atoms are replaced by any halogen.

[0053] As used herein, the term "aromatic ring" refers to a monocyclic or polycyclic carbon ring having 6 to 20 carbon atoms, at least one of which is aromatic. When one of the rings is non-aromatic, the group may be attached through either the aromatic ring or the non-aromatic ring. Examples of aryl groups include, but are not limited to, phenyl, naphthyl, tetrahydronaphthyl, indanyl, biphenyl, phenanthrenyl, anthracenyl, and acenaphthenyl. The term "aromatic ring" may be used interchangeably with the term "aryl group."

[0054] As used herein, the term "treatment" refers to any agent used to obtain a desired pharmacological and / or physiological effect. The effect may be preventive in terms of completely or partially preventing a disease or its symptoms, and / or therapeutic in terms of partially or completely curing a disease and / or the adverse effects caused by the disease. "Treatment" as used herein covers diseases in mammals, particularly humans, and includes: (a) preventing the occurrence of a disease or condition in individuals who are susceptible to the disease but have not yet been diagnosed with the disease; (b) inhibiting the disease, such as arresting the progression of the disease; or (c) alleviating the disease, such as alleviating the symptoms associated with the disease. "Treatment" as used herein covers any medication that administers a drug or compound to an individual to treat, cure, alleviate, improve, reduce or inhibit the individual's disease, including but not limited to administering a drug containing a compound described herein to an individual in need.

[0055] As used herein, the term "cancer" or "tumor" may be any unregulated cell growth. Exemplary cancers include bladder cancer, nasopharyngeal cancer, glioblastoma, non-small cell lung cancer, papillary thyroid cancer, glioblastoma multiforme, colon cancer, rectal cancer, lung cancer, head and neck cancer, kidney cancer, breast cancer, ovarian cancer, liver cancer, bile duct cancer or sarcoma, acute myeloid leukemia, large cell neuroendocrine carcinoma, neuroblastoma, prostate cancer, neuroblastoma, pancreatic cancer, melanoma, head and neck squamous cell carcinoma, cervical cancer, skin cancer, glioma, esophageal cancer, oral squamous cell carcinoma, or gastric cancer, among others.

[0056] Detailed description of the compounds with SIRT6 agonist activity and their uses of the present invention

[0057] The present invention provides a class of compounds with SIRT6 agonist activity and uses thereof, which are described in detail below.

[0058] Compound

[0059] The present invention provides a class of compounds, which are compounds represented by formula (I) or stereoisomers, tautomers, solvates, and pharmaceutically acceptable salts thereof:

[0060] wherein Ring A is selected from a polyvalent aromatic ring; R is selected from -H, halogen, hydroxyl, amino, carbonyl, C optionally substituted by R1 1～6 Alkyl, optionally substituted by R1C 1～6 Alkoxy, optionally substituted by R1 C 1～6 Haloalkyl or optionally substituted by R1 C 1～6 Halogenated oxyalkyl; R1 is selected from C 1～4 Alkyl, hydroxy, amino, nitro, cyano, oxo, carboxyl, carbonyl.

[0061] The compounds of the present invention have SIRT6 agonist activity and can be used as SIRT6 agonists to effectively prevent and / or treat related diseases or symptoms mediated by SIRT6 protein.

[0062] According to an embodiment of the present invention, ring A is selected from a 5-10 membered aromatic ring; R is selected from -H, halogen, hydroxyl, amino, carbonyl, C optionally substituted by R1 1～6 Alkyl, optionally substituted by R1C 1～6 Alkoxy, optionally substituted by R1 C 1～6 Haloalkyl or optionally substituted by R1 C 1～6 Halogenated oxyalkyl; R1 is selected from C 1～4 Alkyl, hydroxy, amino, nitro, cyano, oxo, carboxyl, carbonyl.

[0063] According to an embodiment of the present invention, ring A is selected from a 5-7 membered aromatic ring; R is selected from -H, halogen, hydroxyl, amino, carbonyl, C optionally substituted by R1 1～6 Alkyl, optionally substituted by R1C 1～6 Alkoxy, optionally substituted by R1 C 1～6 Haloalkyl or optionally substituted by R1 C 1～6 Halogenated oxyalkyl; R1 is selected from C 1～4 Alkyl, hydroxy, amino, nitro, cyano, oxo, carboxyl, carbonyl.

[0064] According to an embodiment of the present invention, ring A is selected from a 5-7 membered aromatic ring; R is selected from -H, halogen, hydroxyl, amino, carbonyl, C optionally substituted by R1 1～5 Alkyl, optionally substituted by R1C 1～5 Alkoxy, optionally substituted by R1 C1～5 Haloalkyl or optionally substituted by R1 C 1～5 Halogenated oxyalkyl; R1 is selected from C 1～4 Alkyl, hydroxy, amino, nitro, cyano, oxo, carboxyl, carbonyl.

[0065] According to an embodiment of the present invention, ring A is selected from a 5-7 membered aromatic ring; R is selected from -H, halogen, hydroxyl, amino, carbonyl, C optionally substituted by R1 1～4 Alkyl, optionally substituted by R1C 1～4 Alkoxy, optionally substituted by R1 C 1～4 Haloalkyl or optionally substituted by R1 C 1～4 Halogenated oxyalkyl; R1 is selected from C 1～4 Alkyl, hydroxy, amino, nitro, cyano, oxo, carboxyl, carbonyl.

[0066] According to an embodiment of the present invention, ring A is selected from a 5-7 membered aromatic ring; R is selected from -H, halogen, hydroxyl, amino, carbonyl, C optionally substituted by R1 1～3 Alkyl, optionally substituted by R1C 1～3 Alkoxy, optionally substituted by R1 C 1～3 Haloalkyl or optionally substituted by R1 C 1～3 Halogenated oxyalkyl; R1 is selected from C 1～4 Alkyl, hydroxy, amino, nitro, cyano, oxo, carboxyl, carbonyl.

[0067] According to an embodiment of the present invention, ring A is selected from a 5-7 membered aromatic ring; R is selected from -H, halogen, hydroxyl, amino, carbonyl, C optionally substituted by R1 1～3 Alkyl, optionally substituted by R1C 1～3 Alkoxy, optionally substituted by R1 C 1～3 Haloalkyl or optionally substituted by R1 C 1～3 Halogenated oxyalkyl; R1 is selected from C 1～3 Alkyl, hydroxy, amino, nitro, cyano, oxo, carboxyl, carbonyl.

[0068] According to an embodiment of the present invention, ring A is selected from a six-membered aromatic ring.

[0069] According to an embodiment of the present invention, R is selected from H, halogen or C 1～6 Alkoxy.

[0070] According to an embodiment of the present invention, ring A is selected from a benzene ring.

[0071] According to an embodiment of the present invention, R is selected from C 1～6 Alkoxy.

[0072] According to an embodiment of the present invention, R is selected from C 1～5 Alkoxy.

[0073] According to an embodiment of the present invention, R is selected from C 1～4 Alkoxy.

[0074] According to an embodiment of the present invention, R is selected from C 1～3 Alkoxy.

[0075] According to an embodiment of the present invention, the compound represented by formula (I) has a structure represented by the following formula (II):

[0076] According to an embodiment of the present invention, the compound represented by formula (I) has the structure shown below:

[0077] Pharmaceutical composition

[0078] The present invention provides a pharmaceutical composition comprising the aforementioned compound, or a stereoisomer, tautomer, solvate, or pharmaceutically acceptable salt thereof. As previously mentioned, the aforementioned compound exhibits SIRT6 agonist activity. Therefore, the pharmaceutical composition of the present invention can promote the expression or activity of SIRT6 protein, thereby effectively preventing and / or treating diseases or symptoms mediated by SIRT6 protein.

[0079] According to an embodiment of the present invention, the pharmaceutical composition further includes a pharmaceutically acceptable carrier or excipient.

[0080] Administration of the pharmaceutical compositions of the present invention can be carried out by any acceptable mode of administration. The pharmaceutical compositions of the present invention can be formulated into formulations including, but not limited to, solid, semisolid, liquid or gaseous forms, such as tablets, capsules, injections, lyophilized powders, and the current methods for preparing these dosage forms are known or will be apparent to those skilled in the art. Typical routes of administration of such pharmaceutical compositions include, but are not limited to, oral, topical, transdermal, inhalation, parenteral, sublingual, buccal, rectal, vaginal and intranasal routes. The term parenteral as used herein includes subcutaneous injection, intravenous, intramuscular, intradermal, intrasternal injection or infusion techniques. The pharmaceutical compositions of the present invention are formulated so as to allow the biologically active ingredients contained therein to be bioavailable after administration of the composition to a patient.

[0081] As used herein, "pharmaceutically acceptable carrier" includes, but is not limited to, any adjuvant, carrier, excipient, glidant, sweetener, diluent, preservative, dye / colorant, flavoring, surfactant, wetting agent, dispersant, suspending agent, stabilizer, isotonic agent, solvent or emulsifier approved by the relevant governmental regulatory authorities as acceptable for use by humans or livestock.

[0082] As used herein, the term "pharmaceutically acceptable excipient" includes any solvent suitable for the particular intended dosage form. Except to the extent that any conventional excipient is incompatible with the compounds of the present disclosure, such as by producing any adverse biological effects or interacting in a deleterious manner with any other component of the pharmaceutically acceptable composition, their use is also contemplated by the present disclosure.

[0083] use

[0084] The present invention provides the use of the aforementioned compounds as SIRT6 agonists. As previously mentioned, the aforementioned compounds have SIRT6 agonist activity and, when used as SIRT6 agonists, can promote the expression or activity of SIRT6 protein, or effectively prevent and / or treat diseases or symptoms mediated by SIRT6 protein.

[0085] The present invention provides the use of the aforementioned compounds in the preparation of reagents for promoting the expression or activity of SIRT6 protein. As previously mentioned, the aforementioned compounds have SIRT6 agonist activity and can be used as reagents to effectively promote the expression or activity of SIRT6 protein. They are particularly useful for increasing the expression or activity of SIRT6 protein in cells in vitro, for example, to construct desired cell models.

[0086] The present invention provides the use of the aforementioned compound or pharmaceutical composition in the preparation of a medicament for preventing and / or treating diseases or symptoms mediated by the SIRT6 protein. As previously mentioned, the aforementioned compound has SIRT6 agonist activity and, as a SIRT6 medicament, can effectively prevent and / or treat diseases or symptoms mediated by the SIRT6 protein.

[0087] The present invention provides the aforementioned compounds or pharmaceutical compositions for use in preventing and / or treating diseases or symptoms mediated by SIRT6 protein. As can be seen from the foregoing, the aforementioned compounds have SIRT6 agonist activity and can effectively prevent and / or treat diseases or symptoms mediated by SIRT6 protein.

[0088] According to an embodiment of the present invention, the related diseases or symptoms mediated by the SIRT6 protein include but are not limited to aging-related diseases, cancer, inflammation, and anti-aging.

[0089] According to an embodiment of the present invention, the aging-related diseases include but are not limited to osteoarthritis, intervertebral disc herniation, osteoporosis, Alzheimer's disease, atherosclerosis, liver fibrosis, age-related macular degeneration, diabetic nephropathy, and idiopathic pulmonary fibrosis.

[0090] It should be noted that the cancer described in the present invention can inhibit its development or growth by activating or promoting the expression of SIRT6 protein. According to an embodiment of the present invention, the cancer is selected from but not limited to bladder cancer, nasopharyngeal carcinoma or glioblastoma.

[0091] In this article, "anti-aging" refers to the ability to prevent the aging of an individual organism, such as anti-skin aging, anti-cell aging, or delaying the shortening of telomere length.

[0092] method

[0093] The present invention provides a method for activating and / or promoting SIRT6 protein activity in cells in vitro. According to an embodiment of the present invention, the method comprises contacting the cells with the aforementioned compound. As previously mentioned, the aforementioned compound has SIRT6 agonist activity. Thus, contacting the aforementioned compound with cells can activate and / or promote the expression or activity of SIRT6 protein in the cells.

[0094] The present invention provides a method for preventing and / or treating diseases or symptoms mediated by the SIRT6 protein. According to an embodiment of the present invention, the method comprises administering a pharmaceutically acceptable amount of the aforementioned compound or pharmaceutical composition to a subject. As previously mentioned, the aforementioned compound exhibits SIRT6 agonist activity. Thus, the aforementioned compound can effectively prevent and / or treat diseases or symptoms mediated by the SIRT6 protein.

[0095] The effective amount of the compounds described herein may vary depending on the mode of administration and the severity of the disease to be treated. The preferred effective amount can be determined by one of ordinary skill in the art based on various factors (e.g., through clinical trials). These factors include, but are not limited to, the pharmacokinetic parameters of the active ingredient, such as bioavailability, metabolism, and half-life; the severity of the disease to be treated, the patient's weight, the patient's immune status, and the route of administration. For example, depending on the exigencies of the treatment, several divided doses may be administered daily, or the dose may be reduced proportionally.

[0096] The compounds of the present invention can be incorporated into drugs suitable for parenteral administration (e.g., intravenous, subcutaneous, intraperitoneal, intramuscular). These drugs can be prepared in various forms, such as liquid, semi-solid and solid dosage forms.

[0097] According to an embodiment of the present invention, the related diseases or symptoms mediated by the SIRT6 protein include but are not limited to aging-related diseases, cancer, inflammation, and anti-aging.

[0098] According to an embodiment of the present invention, the aging-related diseases include but are not limited to osteoarthritis, intervertebral disc herniation, osteoporosis, Alzheimer's disease, atherosclerosis, liver fibrosis, age-related macular degeneration, diabetic nephropathy, and idiopathic pulmonary fibrosis.

[0099] It should be noted that the cancer described in the present invention can inhibit its development or growth by activating or promoting the expression of SIRT6 protein. According to an embodiment of the present invention, the cancer is selected from but not limited to bladder cancer, nasopharyngeal carcinoma or glioblastoma.

[0100] Below, the scheme of the present invention will be explained in conjunction with embodiment.It will be understood by those skilled in the art that the following examples are only used to illustrate the present invention and should not be regarded as limiting the scope of the present invention.In the embodiment, if specific technology or conditions are not indicated, the technology or conditions described in the literature in this area or the product instructions are used.The reagents or instruments used are not indicated by the manufacturer, and are all conventional products that can be obtained by commercial purchase.

[0101] Example 1: Acquisition of Compound 1

[0102] Compound 1 was synthesized by Shanghai TOPScience Technology Co., Ltd. The hydrogen spectrum of compound 1 is shown in Figure 1. Its hydrogen spectrum is characterized by:

[0103] 1 H NMR(400MHz,Chloroform-d)δ7.95(s,1H),7.57(s,1H),7.40(s,1H),7.38-7.31(m,3H),7.22(d, 2H),6.61(s,1H),6.44(s,1H),6.20(s,1H),5.28(s,2H),4.51(s,2H),4.47(d,2H),3.46(s,3H).

[0104] The structure of compound 1 is shown below:

[0105] Example 2: Activity Detection of Compound 1

[0106] 1. Verification of SIRT6 enzyme activity of compound 1

[0107] First, a lysine long-chain polypeptide (RHKK-Lys(Ac)-AMC) synthesized by the Fmoc solid-phase synthesis method was acetylated on the last lysine and connected to coumarin. Coumarin is a natural fluorescent group that emits fluorescence at a wavelength of 460nm only when it is in a free state and excited at a wavelength of 360nm. Then, a compound is added to the system containing the lysine long-chain polypeptide, and the compound is allowed to catalyze the removal of the lysine acetylation modification by SIRT6 protein in the system, thereby exposing the coumarin. In the subsequent operation of adding trypsin, the chemical bond connecting coumarin to lysine will be cut, thereby producing free coumarin that can emit a fluorescent signal. The activity of SIRT6 deacetylase in the system is reflected by detecting the strength of the fluorescent signal, thereby reflecting the activation effect of the added compound on the SIRT6 enzyme activity. Among them, the detailed steps are as follows:

[0108] 1) In vitro enzyme activity assay of Compound 1 in Example 1 was performed using a 384-well plate with a transparent bottom and black inner wall.

[0109] 2) The reaction system for enzyme activity detection is shown in Table 1:

[0110] Table 1

[0111] Add the test compound to a final concentration of 50 μM and then add the reaction system in Table 1. The reaction reagents in Table 1 need to be diluted to the final concentration with a buffer solution. The composition of the buffer solution is shown in Table 2:

[0112] Table 2

[0113] The total volume was made up to 20 μl with buffer solution and incubated in a 37°C constant temperature incubator in the dark for 2 hours.

[0114] 3) After the reaction is complete, add 40 mM NAM and 6 mg / ml trypsin per 20 μl of the reaction system and mix the system thoroughly at 160 rpm in a 25°C incubator for 30 minutes.

[0115] 4) Set the excitation wavelength of the microplate reader to 360 nm and the emission wavelength to 460 nm, test the reaction system, and calculate the catalytic activity of SIRT6 according to the following formula:

[0116] The detection results of compound 1 are shown in Figure 2. The results show that compared with the existing SIRT6 agonist MDL-800 and the natural compound ellagic acid that has been reported to have SIRT6 enzyme activity agonist effect, compound 1 has a higher SIRT6 enzyme activity activation effect.

[0117] 2. Compound dose-effect curve detection

[0118] Compound 1 from Example 1 was used to establish different concentration gradients. The compound at each concentration was detected according to the fluorescence reaction system of step 2) to draw a dose-effect curve. The detection results of compound 1 are shown in Figure 3. The results showed that compared with the existing SIRT6 enzyme activity agonist MDL-800, compound 1 had a better agonist effect in the in vitro enzyme activity detection system, and the log (IC50) of compound 1 was 1.988.

[0119] 3. Compound Toxicity Testing

[0120] Compound 1 prepared in Example 1 was prepared into different concentration gradients and then added to the culture supernatant of the mouse hippocampal neuronal cell line HT22. After culturing at 37°C for 48 hours, CCK8 was used to detect the proliferation inhibition of the cells at each concentration. The detection results of compound 1 are shown in Figure 4. The results showed that compound 1 has strong cytotoxicity and can be used as a potential chemotherapy drug. Since the activity of SIRT6 can affect the efficacy of cancer treatment, the compounds of the present invention have strong cytotoxicity and can significantly activate SIRT6, which is expected to improve the efficacy of cancer chemotherapy and reduce the risk of cancer recurrence and metastasis after chemotherapy.

[0121] 4. Detection of compound physiological functions

[0122] Compound 1 from Example 1 was added to the culture supernatant of the mouse hippocampal neuronal cell line HT22 at varying concentrations. After incubation at 37°C for 48 hours, proteins from the 48-hour-treated mouse hippocampal neuronal cell line HT22 were collected and Western blot analysis was performed for acetylation at the H3K9 site. The activation effect of Compound 1 on the catalytic effect of SIRT6 in the cell lines was evaluated. The detection results of Compound 1 are shown in Figure 5. The results showed that Compound 1 effectively enhanced the deacetylation activity of SIRT6 on histones in both cell lines.

[0123] Example 3: Verification of the anti-aging activity of compounds at the cellular level

[0124] Considering that cell lines have the characteristics of immortalized cells and cannot well characterize the characteristics of senescent cells, the experiments in this example were conducted in primary cells or the C2C12 cell line specifically used for studying senescence.

[0125] Currently, common indicators for detecting cell senescence levels in academia include: SA-β-Gal staining and detection of p16 and p21 senescence-related gene expression. The specific steps are as follows:

[0126] 1) β-galactose was used to induce senescence in the C2C12 cell line. Compound 1 from Example 1 was added to the culture medium and co-cultured with the cells for 24 hours. RNA was extracted from the cells and reverse-transcribed into cDNA. The expression of the p16 and p21 genes in the cells was detected by quantitative PCR.

[0127] 2) β-galactose was used to induce senescence in the C2C12 cell line. Compound 1 from Example 1 was added to the culture medium and co-cultured with the cells for 24 hours. The cells were stained with SA-β-Gal to examine the effect of Compound 1 on the SA-β-Gal staining positivity rate.

[0128] 3) Primary cells were induced to undergo replicative senescence by multiple passages. Compound 1 from Example 1 was added to the culture medium of the primary cells and co-cultured with the cells for 24 hours. RNA was extracted from the cells, reverse transcribed into cDNA, and the expression of the p16 and p21 genes in the cells was detected by fluorescent quantitative PCR.

[0129] 4) Primary cells were induced to undergo replicative senescence by multiple passages. Compound 1 from Example 1 was added to the culture medium of the primary cells and co-cultured with the cells for 24 hours. The primary cells were stained with SA-β-Gal to examine the effect of compound addition on the SA-β-Gal staining positivity rate.

[0130] The results showed that the intervention of compound 1 reduced the SA-β-Gal staining positivity rate or the expression levels of p16 and p21 genes in primary cells of induced senescent C2C12 that had reached the division limit through replicative senescence, further demonstrating that the compounds of the present invention have anti-aging activity at the cellular level.

[0131] In the description of this specification, the reference terms "one embodiment", "some embodiments", "example", "specific example", or "some examples" mean that the specific features, structures, materials or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the schematic representations of the above terms do not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described can be combined in any one or more embodiments or examples in a suitable manner. In addition, those skilled in the art can combine and combine different embodiments or examples described in this specification and features of different embodiments or examples without contradiction.

[0132] Although the embodiments of the present invention have been shown and described above, it will be understood that the above embodiments are illustrative and are not to be construed as limitations on the present invention. A person skilled in the art may change, modify, replace and modify the above embodiments within the scope of the present invention.

Claims

1. A compound, which is a compound represented by formula (I) or a stereoisomer, tautomer, solvate, or pharmaceutically acceptable salt thereof: in, Ring A is selected from polyvalent aromatic rings; R is selected from -H, halogen, hydroxy, amino, carbonyl, C optionally substituted by R1 1～6 Alkyl, optionally substituted by R1C 1～6 Alkoxy, optionally substituted by R1C 1～6 haloalkyl or optionally substituted by R1C 1～6 Halogenated oxyalkyl; R1 is selected from C 1～4 Alkyl, hydroxy, amino, nitro, cyano, oxo, carboxyl, carbonyl.

2. The compound according to claim 1, characterized in that Ring A is selected from a six-membered aromatic ring; R is selected from H, halogen or C 1～6 Alkoxy.

3. The compound according to claim 1, characterized in that Ring A is selected from a benzene ring; R is selected from C 1～6 Alkoxy.

4. The compound according to claim 1, characterized in that The compound represented by formula (I) has the structure shown below:

5. A pharmaceutical composition, characterized in that The pharmaceutical composition comprises: the compound according to any one of claims 1 to 4, or its stereoisomers, tautomers, solvates, and pharmaceutically acceptable salts.

6. The pharmaceutical composition according to claim 5, characterized in that The pharmaceutical composition further comprises a pharmaceutically acceptable carrier or excipient.

7. Use of the compound according to any one of claims 1 to 4 in the preparation of an agent for activating and / or promoting the expression or activity of SIRT6 protein.

8. Use of the compound according to any one of claims 1 to 4 or the pharmaceutical composition according to claim 5 in the preparation of a medicament for preventing and / or treating diseases or symptoms mediated by SIRT6 protein.

9. Use of the compound according to any one of claims 1 to 4 or the pharmaceutical composition according to claim 5 for preventing and / or treating diseases or symptoms mediated by SIRT6 protein.

10. The use according to claim 8 or 9, characterized in that The SIRT6 protein-mediated related diseases or symptoms include aging-related diseases, cancer, inflammation or anti-aging.

11. The use according to claim 10, characterized in that The aging-related diseases include osteoarthritis, intervertebral disc herniation, osteoporosis, Alzheimer's disease, atherosclerosis, liver fibrosis, age-related macular degeneration, diabetic nephropathy, and idiopathic pulmonary fibrosis.

12. A method for activating and / or promoting the activity of SIRT6 protein in vitro, characterized in that: include: The cell is contacted with the compound according to any one of claims 1 to 4.

13. A method for preventing and / or treating SIRT6 protein-mediated diseases or symptoms, characterized in that: include: A pharmaceutically acceptable dose of the compound according to any one of claims 1 to 4 or the pharmaceutical composition according to claim 5 is administered to a subject.

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