Compound having sirt6 agonistic activity and use thereof

Abstract

Provided is a compound. The compound is a compound represented by formula (I) or a stereoisomer, tautomer, solvate, or 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 202311679118.7 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 belongs to the field of biopharmaceutical technology, and specifically relates to a compound with SIRT6 agonist activity and uses thereof. Background Art

[0004] SIRT6, a histone deacetylase, is a key protein that regulates critical biological processes such as DNA repair, cellular metabolism, and apoptosis through epigenetic modification. SIRT6, also known as the "longevity protein," is a key drug target for age-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 age-related diseases, and achieving healthy aging.

[0005] Although several agonists targeting SIRT6 deacetylase, such as MDL-800 and MDL-811, have been shown to have anti-inflammatory effects, screening for SIRT6 agonists with lower toxicity or better efficacy remains of great scientific value. Therefore, the development of drugs with enhanced SIRT6 agonist activity is urgently needed. Summary of the Invention

[0006] 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 compound having SIRT6 agonist activity and its use, wherein the compound has SIRT6 agonist activity and can be used as a SIRT6 agonist to effectively prevent and / or treat diseases or symptoms mediated by the SIRT6 protein.

[0007] In the first aspect of the present invention, the present invention provides a compound, which is a compound represented by formula (I) or a stereoisomer, tautomer, solvate, or pharmaceutically acceptable salt thereof:

[0008] Wherein, R1 and R2 are each independently selected from H, halogen, C 1～6 Alkyl, C 1～6 Oxyalkyl, C 1～6 Haloalkyl or C 1～6 Halogenated oxyalkyl groups;

[0009] Each X1 is independently selected from C, O, S or N;

[0010] n is any integer between 1 and 6.

[0011] 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.

[0012] According to an embodiment of the present invention, R1 is selected from C 1～6 Alkyl, C 1～6 Oxyalkyl, C 1～6 Haloalkyl or C 1～6 Halogenated oxyalkyl.

[0013] According to an embodiment of the present invention, R1 is selected from C 1～3 Alkyl or C 1～3 Oxyalkyl.

[0014] According to an embodiment of the present invention, R2 is selected from H, halogen or C 1～6 alkyl.

[0015] According to an embodiment of the present invention, R2 is selected from halogen.

[0016] According to an embodiment of the present invention, each X1 is independently selected from C or S.

[0017] According to an embodiment of the present invention, n is 1, 2, 3 or 4.

[0018] According to an embodiment of the present invention, n is 1, 2 or 3.

[0019] According to an embodiment of the present invention, the compound represented by formula (I) has a structure represented by the following formula (II):

[0020] According to an embodiment of the present invention, the compound represented by formula (I) has the structure shown below:

[0021] In a second aspect, the present invention provides a pharmaceutical composition. According to an embodiment of the present invention, the pharmaceutical composition includes the compound described in the first aspect. As previously mentioned, the compound described in the first aspect has SIRT6 agonist activity and can act as a SIRT6 agonist. Thus, the pharmaceutical composition containing the compound can effectively prevent and / or treat diseases or symptoms mediated by the SIRT6 protein.

[0022] According to an embodiment of the present invention, the pharmaceutical composition further includes a pharmaceutically acceptable excipient.

[0023] 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 promote the expression or activity of SIRT6 protein in cells, or effectively prevent and / or treat diseases or symptoms mediated by SIRT6 protein.

[0024] In a fourth aspect, the present invention provides the use of the compound described in the first aspect in preparing a reagent for promoting the expression or activity of a SIRT6 protein. As previously mentioned, the compound described in the first aspect has SIRT6 agonist activity and, when used as a reagent, can effectively promote the expression or activity of a SIRT6 protein, particularly increasing SIRT6 protein expression or activity in cells in vitro, for example, to construct a desired cell model.

[0025] In a fifth aspect, the present invention provides the compound of the first aspect or the pharmaceutical composition of the second aspect for use in preventing and / or treating diseases or symptoms mediated by SIRT6 protein, or for use in preventing and / or treating diseases or symptoms mediated by SIRT6 protein, and / or for preparing a medicament for preventing and / or treating diseases or symptoms mediated by SIRT6 protein. As mentioned above, the compound of the first aspect has SIRT6 agonist activity and, as a SIRT6 drug, can effectively prevent and / or treat diseases or symptoms mediated by SIRT6 protein.

[0026] According to an embodiment of the present invention, the related diseases or symptoms mediated by the SIRT6 protein include aging-related diseases, cancer, inflammation, and anti-aging.

[0027] According to an embodiment of the present invention, 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.

[0028] In a sixth 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 compound with cells can promote the expression or activity of SIRT6 protein in the cells.

[0029] In a seventh 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 of the first aspect or the pharmaceutical composition of the second aspect to a subject. As previously mentioned, the compound of the first aspect has SIRT6 agonist activity. Thus, the above-mentioned compound can effectively prevent and / or treat diseases or symptoms mediated by the SIRT6 protein.

[0030] 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

[0031] 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:

[0032] FIG1 is a hydrogen spectrum of compound 1 in Example 1;

[0033] FIG2 is a hydrogen spectrum of compound 2 in Example 1;

[0034] FIG3 is the detection result of SIRT6 enzyme activity of compound 1 in Example 2;

[0035] FIG4 is the detection result of SIRT6 enzyme activity of compound 2 in Example 2;

[0036] FIG5 is the test result of the dose-effect curve of compound 1 in Example 2;

[0037] FIG6 is the test result of the dose-effect curve of compound 2 in Example 2;

[0038] FIG7 shows the toxicity test results of Compound 1 using CCK-8 in Example 2;

[0039] FIG8 is the toxicity test result of compound 2 using CCK-8 in Example 2;

[0040] FIG9 is the detection result of Western blot detection of the activation effect of compound 1 on SIRT6 protein activity in cell lines in Example 2;

[0041] FIG10 is the detection result of Western blot detection of the activation effect of compound 2 on SIRT6 protein activity in cell lines in Example 2. DETAILED DESCRIPTION

[0042] 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.

[0043] 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.

[0044] Detailed description of the invention

[0045] Definitions and General Terms

[0046] 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.

[0047] 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.

[0048] 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.

[0049] 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.

[0050] 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.

[0051] 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 invention are within the scope of the invention.

[0052] 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.

[0053] 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.

[0054] 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.

[0055] As used herein, the term "halogen" refers to a fluorine, chlorine, bromine or iodine atom.

[0056] 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.

[0057] As used herein, the term "haloalkyl" refers to an alkyl group in which one or more H atoms are replaced by any halogen.

[0058] 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-C6 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").

[0059] As used herein, the term "halooxyalkyl" refers to an oxyalkyl group in which one or more H atoms are replaced by any halogen.

[0060] As used herein, the term "pharmaceutically acceptable excipient" may include any solvent, solid excipient, diluent or other liquid excipient, etc., suitable for the specific target dosage form. Except to the extent that any conventional excipient is incompatible with the compound of the present invention, such as any adverse biological effect produced or interaction with any other component of the pharmaceutically acceptable composition in a harmful manner, their use is also contemplated by the present invention.

[0061] 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.

[0062] As used herein, the term "cancer" or "tumor" may be any unregulated cell growth. Exemplary cancers include non-small cell lung cancer, papillary thyroid cancer, glioblastoma multiforme, colon cancer, rectal cancer, lung cancer, head and neck cancer, kidney cancer, bladder cancer, nasopharyngeal cancer, glioblastoma, 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.

[0063] Detailed description of the compounds with SIRT6 agonist activity and their uses of the present invention

[0064] The present invention provides a compound with SIRT6 agonist activity and its use, which will be described in detail below.

[0065] Compound

[0066] In the first aspect of the present invention, the present invention provides a compound, which is a compound represented by formula (I) or a stereoisomer, tautomer, solvate, or pharmaceutically acceptable salt thereof:

[0067] Wherein, R1 and R2 are each independently selected from H, halogen, C 1～6 Alkyl, C 1～6 Oxyalkyl, C 1～6 Haloalkyl or C 1～6 Halogenated oxyalkyl groups;

[0068] Each X1 is independently selected from C, O, S or N;

[0069] n is any integer between 1 and 6.

[0070] 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.

[0071] According to an embodiment of the present invention, R1 is selected from C 1～6 Alkyl, C 1～6 Oxyalkyl, C 1～6 Haloalkyl or C 1～6 Halogenated oxyalkyl.

[0072] According to an embodiment of the present invention, R1 is selected from C 1～3 Alkyl or C 1～3 Oxyalkyl.

[0073] According to an embodiment of the present invention, R1 is C 1～3 Oxyalkyl.

[0074] According to an embodiment of the present invention, R1 is -O-CH3.

[0075] According to an embodiment of the present invention, R2 is selected from H, halogen or C 1～6 alkyl.

[0076] According to an embodiment of the present invention, R2 is selected from halogen.

[0077] According to an embodiment of the present invention, R2 is Cl.

[0078] According to an embodiment of the present invention, each X1 is independently selected from C or S.

[0079] According to an embodiment of the present invention, X1 is C.

[0080] According to an embodiment of the present invention, X1 is S.

[0081] According to an embodiment of the present invention, n is 1, 2, 3 or 4.

[0082] According to an embodiment of the present invention, n is 1, 2 or 3.

[0083] According to an embodiment of the present invention, n is 1.

[0084] According to an embodiment of the present invention, n is 2.

[0085] According to an embodiment of the present invention, n is 3.

[0086] According to an embodiment of the present invention, the compound represented by formula (I) has the structure described by the following formula (II):

[0087] According to an embodiment of the present invention, the compound represented by formula (I) has a structure as described in formula (IIa), formula (IIb) or formula (IIc):

[0088] According to an embodiment of the present invention, the compound represented by formula (I) has the structure shown below:

[0089] use

[0090] In a second aspect, the present invention provides a pharmaceutical composition. According to an embodiment of the present invention, the pharmaceutical composition includes the compound described in the first aspect. As previously mentioned, the compound described in the first aspect has SIRT6 agonist activity and can act as a SIRT6 agonist. Thus, the pharmaceutical composition containing the compound can effectively prevent and / or treat diseases or symptoms mediated by the SIRT6 protein.

[0091] According to an embodiment of the present invention, the pharmaceutical composition further includes a pharmaceutically acceptable excipient.

[0092] 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 solid, semisolid, liquid or gaseous formulations, 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.

[0093] 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 promote the expression or activity of SIRT6 protein in cells, or effectively prevent and / or treat diseases or symptoms mediated by SIRT6 protein.

[0094] In a fourth aspect, the present invention provides the use of the compound described in the first aspect in preparing a reagent for promoting the expression or activity of a SIRT6 protein. As previously mentioned, the compound described in the first aspect has SIRT6 agonist activity and, when used as a reagent, can effectively promote the expression or activity of a SIRT6 protein, particularly by increasing SIRT6 protein expression or activity in cells in vitro, thereby constructing a desired cell model.

[0095] In the fifth aspect of the present invention, the present invention provides 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 SIRT6 protein.

[0096] The present invention provides the use of the compound described in the first aspect or the pharmaceutical composition described in the second aspect in preventing and / or treating diseases or symptoms mediated by SIRT6 protein.

[0097] The present invention proposes the use of the compound described in the first aspect or the pharmaceutical composition described in the second aspect in the preparation of a drug for preventing and / or treating diseases or symptoms mediated by SIRT6 protein.

[0098] As can be seen from the foregoing, the compound described in the first aspect has SIRT6 agonist activity, and can be used as a SIRT6 drug to effectively prevent and / or treat diseases or symptoms mediated by SIRT6 protein.

[0099] According to an embodiment of the present invention, the related diseases or symptoms mediated by the SIRT6 protein include aging-related diseases, cancer, inflammation, and anti-aging.

[0100] According to an embodiment of the present invention, 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.

[0101] 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.

[0102] Method for activating cell SIRT6 protein activity in vitro

[0103] In a sixth 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 compound with cells can promote the expression or activity of SIRT6 protein in the cells.

[0104] In a seventh 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 of the first aspect or the pharmaceutical composition of the second aspect to a subject. As previously mentioned, the compound of the first aspect has SIRT6 agonist activity. Thus, the above-mentioned compound can effectively prevent and / or treat diseases or symptoms mediated by the SIRT6 protein.

[0105] 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.

[0106] 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.

[0107] According to an embodiment of the present invention, the related diseases or symptoms mediated by the SIRT6 protein include aging-related diseases, cancer, inflammation, and anti-aging.

[0108] According to an embodiment of the present invention, 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.

[0109] In some optional embodiments of the present invention, the cancer is bladder cancer, nasopharyngeal carcinoma, or malignant glioma.

[0110] 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.

[0111] Example 1: Acquisition of Compounds

[0112] 1. 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:

[0113] 1 H NMR (400MHz, DMSO-d6) δ12.37(s,1H),9.80(s,1H),8.10(s,1H),7.74(d,1H),7.65(d,1H),7.40(d,1H),7.31( d,1H),7.06(d,1H),3.80(s,3H),3.71-3.60(m,2H),2.43-2.21(m,3H),1.92-1.81(m,2H),1.73-1.58(m,2H).

[0114] The structural formula of compound 1 is shown below:

[0115] 2. Compound 2 was synthesized by Shanghai TOPScience Technology Co., Ltd. The hydrogen spectrum of compound 2 is shown in Figure 2. Its hydrogen spectrum is characterized by:

[0116] 1H NMR(400MHz,DMSO-d6)δ9.93(s,1H),9.80(s,1H),7.74(d,1H),7.62(dd,2H),7.39(dd,1H),7.17(d,1H),7.08 (d,1H),3.80(s,3H),3.71-3.62(m,2H),2.81(t,2H),2.40-2.11(m,7H),1.91-1.80(m,2H),1.70-1.56(m,2H).

[0117] The structural formula of compound 2 is shown below:

[0118] Example 2: Activity detection of compounds

[0119] 1. Verification of SIRT6 enzyme activity of compounds

[0120] First, a lysine long-chain polypeptide (RHKK-Lys(Ac)-AMC) synthesized by the Fmoc solid-phase peptide synthesis method was acetylated on the last lysine and linked 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:

[0121] 1) In vitro enzyme activity assays were performed on Compound 1 and Compound 2 in Example 1 using a well plate with a transparent bottom and a black inner wall.

[0122] 2) The reaction system for enzyme activity detection is shown in Table 1:

[0123] Table 1

[0124] Add the test compound to a final concentration of 50 μM and then add the above reaction system. The reaction reagents in the above table need to be diluted with a buffer solution to the final concentration. The buffer solution ratio is shown in Table 2:

[0125] Table 2

[0126] 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.

[0127] 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.

[0128] 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:

[0129] The detection results of compound 1 are shown in Figure 3, and the detection results of compound 2 are shown in Figure 4. The results showed that compared with the existing agonists MDL-800 and ellagic acid, both compound 1 and compound 2 have higher activation effects on SIRT6 enzyme activity.

[0130] 2. Compound dose-effect curve detection

[0131] Compound 1 and compound 2 in Example 1 were taken and different concentration gradients were set up. The compounds at each concentration were tested according to the fluorescence reaction system in step 2, and a dose-effect curve was drawn. The test results of compound 1 are shown in Figure 5, and the test results of compound 2 are shown in Figure 6. The results showed that in the in vitro enzyme activity test system, the log (IC 50 ) was 1.881, and the log(IC 50 ) is 1.556.

[0132] 3. Compound toxicity testing

[0133] Compound 1 and Compound 2 from Example 1 were added to the culture supernatant of the mouse hippocampal neuronal cell line HT22 at different concentration gradients. After incubation at 37°C for 48 hours, CCK8 was used to detect the cell proliferation inhibition at each concentration. The detection results for Compound 1 are shown in Figure 7 , and the detection results for Compound 2 are shown in Figure 8 . The results showed that both Compound 1 and Compound 2 had low toxicity. Compound 1 maintained a high cell viability at a concentration of 0.550 mmol / L, and Compound 2 maintained a high cell viability at a concentration of 0.989 mmol / L.

[0134] 4. Detection of compound physiological functions

[0135] Compound 1 and Compound 2 from Example 1 were co-cultured with the mouse hippocampal neuronal cell line HT22 at varying concentrations at 37°C for 48 hours. Protein from the 48-hour treated mouse hippocampal neuronal cell line HT22 was then collected and Western blot assayed for acetylation at the H3K9 site. The activation effects of Compound 1 and Compound 2 on the catalytic activity of SIRT6 in the cell lines were evaluated. The assay results for Compound 1 are shown in Figure 9 , and for Compound 2, in Figure 10 . The results showed that both Compound 1 and Compound 2 effectively enhanced the deacetylation activity of SIRT6 on histones in the cell lines.

[0136] Example 3: Verification of the anti-aging activity of compounds at the cellular level

[0137] 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.

[0138] Currently, the common indicators used in the academic community to detect cell senescence levels include: SA-β-Gal staining and detection of p16 and p21 senescence-related gene expression. The specific steps are as follows:

[0139] 1) β-galactose was used to induce senescence in the C2C12 cell line. Compound 1 and Compound 2 from Example 1 were 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.

[0140] 2) β-galactose was used to induce senescence in the C2C12 cell line. Compound 1 and Compound 2 from Example 1 were 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 the added compounds on the SA-β-Gal staining positivity rate.

[0141] 3) Primary cells were induced to undergo replicative senescence by multiple passages. Compounds 1 and 2 from Example 1 were 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.

[0142] 4) Primary cells were induced to undergo replicative senescence by multiple passages. Compound 1 and Compound 2 from Example 1 were 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.

[0143] The results showed that the intervention of Compound 1 and Compound 2 decreased 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 produced replicative senescence and reached the division limit, further demonstrating that the compounds of the present invention have anti-aging activity at the cellular level.

[0144] 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.

[0145] 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, R1 and R2 are each independently selected from H, halogen, C 1～6 Alkyl, C 1～6 Oxyalkyl, C 1～6 Haloalkyl or C 1～6 Halogenated oxyalkyl; Each X1 is independently selected from C, O, S or N; n is any integer between 1 and 6.

2. The compound according to claim 1, characterized in that R1 is selected from C 1～6 Alkyl, C 1～6 Oxyalkyl, C 1～6 Haloalkyl or C 1～6 Halogenated oxyalkyl; R2 is selected from H, halogen or C 1～6 alkyl; Each X1 is independently selected from C or S; n is 1, 2, 3 or 4.

3. The compound according to claim 1, characterized in that R1 is selected from C 1～3 Alkyl or C 1～3 Oxyalkyl; R2 is selected from halogen; n is 1, 2 or 3.

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

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

6. A pharmaceutical composition, characterized in that The invention comprises the compound according to any one of claims 1 to 5, and optionally a pharmaceutically acceptable auxiliary material.

7. Use of the compound according to any one of claims 1 to 5 as a SIRT6 agonist.

8. Use of the compound according to any one of claims 1 to 5 in the preparation of an agent for promoting the expression or activity of SIRT6 protein.

9. The compound according to any one of claims 1 to 5 or the pharmaceutical composition according to claim 6, for preventing and / or treating diseases or symptoms mediated by SIRT6 protein, or for the following uses: Preventing and / or treating SIRT6 protein-mediated diseases or symptoms, and / or Preparation of drugs for preventing and / or treating diseases or symptoms mediated by SIRT6 protein.

10. The use according to claim 9, characterized in that The SIRT6 protein-mediated related diseases or symptoms include aging-related diseases, cancer, inflammation, and 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 5.

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 5 or the pharmaceutical composition according to claim 6 is administered to a subject.

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