WO2025118982A1 - Compounds having sirt6 agonistic activity and use thereof - Google Patents

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

Priority information

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 the entire text of which is incorporated herein by reference.

Technical Field

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

Background Art

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 ⁺ -dependent histone deacetylase activity, it also has mono-ADP ribosyltransferase and de-fatty acylation functions.

Studies have shown that SIRT6 activity is regulated by multiple factors, mainly through catalyzing the deacetylation of related proteins in cells, and is mainly involved in life activities such as DNA repair, inflammation, and glucose and lipid metabolism. It is a key drug target for aging-related diseases, inflammation, cancer and other diseases. Therefore, the development of drugs with the effect of activating SIRT6 deacetylase activity is of great significance for delaying aging, treating aging-related diseases, and achieving healthy aging.

Although there are many agonists targeting SIRT6 deacetylase activity, such as MDL-800 and MDL-811, which have been shown to have anti-inflammatory effects, screening SIRT6 agonists with lower toxicity and better effects still has very important scientific value. Therefore, it is urgent to develop a drug with better SIRT6 agonist activity.

Summary of the invention

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

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:

Wherein, ring A is selected from a polyvalent aromatic ring; R is selected from H, halogen, hydroxyl, amino, carbonyl, C _1-6 alkyl optionally substituted by R ₁ , C _1-6 alkoxy optionally substituted by R ₁ , C _1-6 haloalkyl optionally substituted by R ₁ or C _1-6 halooxyalkyl optionally substituted by R ₁ ; R ₁ is selected from C _1-4 alkyl, hydroxyl, amino, nitro, cyano, oxo, carboxyl and carbonyl.

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

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

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

According to an embodiment of the present invention, R is selected from H, halogen or C _1-6 alkoxy.

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

According to an embodiment of the present invention, R is selected from C _1-6 alkoxy.

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

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

In the second aspect of the present invention, the present invention proposes a pharmaceutical composition. According to an embodiment of the present invention, the pharmaceutical composition comprises: the compound described in the first aspect, or its stereoisomer, tautomer, solvate, and pharmaceutically acceptable salt. As can be seen from the foregoing, the compound described in the first aspect has SIRT6 agonist activity. Therefore, the pharmaceutical composition described in the present invention can activate and/or promote the expression or activity of SIRT6 protein, thereby effectively preventing and/or treating related diseases mediated by SIRT6 protein.

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

In the third aspect of the present invention, the present invention proposes the use of the compound described in the first aspect as a SIRT6 agonist. As mentioned above, the compound described in the first aspect has SIRT6 agonist activity, and using it as a SIRT6 agonist can activate and/or promote the expression or activity of SIRT6 protein in cells, or effectively prevent and/or treat related diseases mediated by SIRT6 protein.

In the fourth aspect of the present invention, the present invention proposes the use of the compound described in the first aspect in the preparation of a reagent for activating and/or promoting the expression or activity of SIRT6 protein. As mentioned above, the compound described in the first aspect has SIRT6 agonist activity, and using it as a reagent can effectively activate and/or promote the expression or activity of SIRT6 protein, and can be particularly used to increase the expression or activity of SIRT6 protein in in vitro cells, for example, to construct a desired cell model.

In the fifth aspect of the present invention, 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, and the drug is used to prevent and/or treat related diseases or symptoms mediated by SIRT6 protein. As mentioned above, the compound described in the first aspect has SIRT6 agonist activity, and using it as a SIRT6 drug can effectively prevent and/or treat related diseases or symptoms mediated by SIRT6 protein.

In the sixth aspect of the present invention, the present invention proposes the use of the compound described in the first aspect or the pharmaceutical composition described in the second aspect for preventing and/or treating SIRT6 protein-mediated related diseases or symptoms. As mentioned above, the compound described in the first aspect has SIRT6 agonist activity and can effectively prevent and/or treat SIRT6 protein-mediated related diseases or symptoms.

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.

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.

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 cancer.

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

In the eighth aspect of the present invention, the present invention proposes a method for preventing and/or treating related diseases or symptoms mediated by 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 can be seen from the foregoing, the compound described in the first aspect has SIRT6 agonist activity. Thus, the above-mentioned compound can effectively prevent and/or treat related diseases or symptoms mediated by SIRT6 protein.

Additional aspects and advantages of the present invention will be given in part in the following description and in part will be obvious from the following description, or will be learned through practice of the present invention.

BRIEF DESCRIPTION OF THE DRAWINGS

The above and/or additional aspects and advantages of the present invention will become apparent and easily understood from the description of the embodiments in conjunction with the following drawings, in which:

FIG1 is a hydrogen spectrum of compound 1 in Example 1;

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

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

FIG4 is the toxicity test result of compound 1 using CCK-8 in Example 2;

FIG. 5 is the detection result of Western detection of the activation effect of compound 1 on SIRT6 protein activity in cell lines in Example 2.

DETAILED DESCRIPTION

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.

It should be noted that the terms "first" and "second" are used for descriptive purposes only and should not be understood as indicating or implying relative importance or implicitly indicating the number of the indicated technical features. Therefore, the features defined as "first" and "second" may explicitly or implicitly include one or more of the features. Further, in the description of the present invention, unless otherwise specified, the meaning of "plurality" is two or more.

Definitions and general terms

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.

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.

As used herein, the definitions and conventions of stereoisomers are generally in accordance with S. P. 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 also included within the scope of 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 (or centers) in the molecule. The prefixes d and l or (+) and (-) are the signs 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. For a given chemical structure, these stereoisomers are identical except that they are mirror images of each other. 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 may occur when there is no stereoselectivity or stereospecificity in a chemical reaction or process.

Depending on the choice of starting materials and processes, the compounds of the invention may exist in the form of one of the possible isomers or a mixture thereof, for example as pure optical isomers, or as isomer mixtures, such as racemic and diastereomeric mixtures, depending on the number of asymmetric carbon atoms. Optically active (R)- or (S)-isomers may be prepared using chiral synthons or chiral agents, 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.

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.

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

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

In this article, 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 mammals treated therewith. Preferably, the "pharmaceutically acceptable" of the present invention refers to those approved by federal regulatory agencies or national governments or listed in the U.S. Pharmacopoeia or other generally recognized pharmacopeias for use in animals, particularly humans.

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 S.M.Berge et al., describe pharmaceutically acceptable salts in detail in J.Pharmaceutical Sciences, 1977, 66: 1-19.

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

As used herein, the term " _C1-6 alkyl" refers to a straight or branched saturated monovalent hydrocarbon group having 1 _{, 2, 3} , _{4, 5 or 6 carbon atoms, such as C1-5 alkyl, C1-4} alkyl, _C1-3 alkyl, _C2-5 alkyl, _C2-4 alkyl, _C2-3 alkyl. Among _them , 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 _{( -CH2CH2CH2CH2CH3} ), ₂ -pentyl (-CH( _{CH3 ) CH2CH2CH3} ), 3 _- pentyl (-CH( _CH2CH3 ) ₂ ) _, 2-methyl _{- 2} -butyl (-C _{( CH3} ) _{2CH2CH3 )} ), 3-methyl-2-butyl (-CH(CH ₃ )CH(CH ₃ ) ₂ ), 3-methyl-1-butyl (-CH ₂ CH ₂ CH(CH ₃ ) ₂ ), 2-methyl-1-butyl (-CH ₂ CH(CH ₃ )CH ₂ CH ₃ ), n-hexyl (-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CH ₃ ), 2-hexyl (-CH(CH ₃ )CH ₂ CH ₂ CH ₂ CH ₃ ), 3-hexyl (-CH(CH ₂ CH ₃ )(CH ₂ CH ₂ CH ₃ )), 2-methyl-2-pentyl (-C(CH ₃ ) ₂ CH ₂ CH ₂ CH ₃ ), 3-methyl-2-pentyl (-CH(CH ₃ )CH(CH ₃ )CH ₂ CH ₃ ), 4-methyl-2-pentyl (-CH(CH ₃ )CH ₂ CH(CH ₃ ) ₂ ), 3-methyl-3-pentyl (-C(CH ₃ )(CH ₂ CH ₃ ) ₂ ), 2-methyl-3-pentyl (-CH(CH ₂ CH ₃ )CH(CH ₃ ) ₂ ), 2,3-dimethyl-2-butyl (-C(CH ₃ ) ₂ CH(CH ₃ ) ₂ ), 3,3-dimethyl-2-butyl (-CH(CH ₃ )C(CH ₃ ) ₃ ), wherein the alkyl groups may independently be unsubstituted or substituted with one or more substituents described herein.

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

As used herein, the term "C _1-6 alkoxy" refers to a C _1-6 alkyl group having the formula "-O-alkyl", 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 "C 1-6 alkoxy" group 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").

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

As used herein, the term "aromatic ring" refers to a monocyclic or polycyclic carbon ring having 6 to 20 carbon atoms, wherein at least one ring is aromatic. When one of the rings is a non-aromatic ring, the group may be connected via the aromatic ring or via 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".

As used herein, the term "treatment" refers to the use of drugs to obtain the desired pharmacological and/or physiological effects. The effect may be preventive in terms of completely or partially preventing a disease or its symptoms, and/or may be 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 an individual who is susceptible to the disease but has not yet been diagnosed with the disease; (b) inhibiting the disease, such as blocking the progression of the disease; or (c) alleviating the disease, such as alleviating 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 an individual's disease, including but not limited to administering a drug containing a compound described herein to an individual in need.

In this article, the term "cancer" or "tumor" can be any unregulated cell growth. Exemplarily, it can be bladder cancer, nasopharyngeal carcinoma, glioblastoma cancer, 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, etc.

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

The present invention provides a class of compounds with SIRT6 agonist activity and uses thereof, which will be described in detail below.

Compound

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

Wherein, ring A is selected from a polyvalent aromatic ring; R is selected from -H, halogen, hydroxyl, amino, carbonyl, C _1-6 alkyl optionally substituted by R ₁ , C _1-6 alkoxy optionally substituted by R ₁ , C _1-6 haloalkyl optionally substituted by R ₁ or C _1-6 halooxyalkyl optionally substituted by R ₁ ; R ₁ is selected from C _1-4 alkyl, hydroxyl, amino, nitro, cyano, oxo, carboxyl and carbonyl.

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

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 _1-6 alkyl optionally substituted by R ₁ , C _1-6 alkoxy optionally substituted by R ₁ , C _1-6 haloalkyl optionally substituted by R ₁ or C _1-6 halooxyalkyl optionally substituted by R ₁ ; R ₁ is selected from C _1-4 alkyl, hydroxyl, amino, nitro, cyano, oxo, carboxyl, carbonyl.

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 _1-6 alkyl optionally substituted by R ₁ , C _1-6 alkoxy optionally substituted by R ₁ , C _1-6 haloalkyl optionally substituted by R ₁ or C _1-6 halooxyalkyl optionally substituted by R ₁ ; R ₁ is selected from C _1-4 alkyl, hydroxyl, amino, nitro, cyano, oxo, carboxyl, carbonyl.

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 _1-5 alkyl optionally substituted by R ₁ , C _1-5 alkoxy optionally substituted by R ₁ , C _1-5 haloalkyl optionally substituted by R ₁ or C _1-5 halooxyalkyl optionally substituted by R ₁ ; R ₁ is selected from C _1-4 alkyl, hydroxyl, amino, nitro, cyano, oxo, carboxyl, carbonyl.

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 _1-4 alkyl optionally substituted by R ₁ , C _1-4 alkoxy optionally substituted by R ₁ , C _1-4 haloalkyl optionally substituted by R ₁ or C _1-4 halooxyalkyl optionally substituted by R ₁ ; R ₁ is selected from C _1-4 alkyl, hydroxyl, amino, nitro, cyano, oxo, carboxyl, carbonyl.

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 _1-3 alkyl optionally substituted by R ₁ , C _1-3 alkoxy optionally substituted by R ₁ , C _1-3 haloalkyl optionally substituted by R ₁ or C _1-3 halooxyalkyl optionally substituted by R ₁ ; R ₁ is selected from C _1-4 alkyl, hydroxyl, amino, nitro, cyano, oxo, carboxyl, carbonyl.

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 _1-3 alkyl optionally substituted by R ₁ , C _1-3 alkoxy optionally substituted by R ₁ , C _1-3 haloalkyl optionally substituted by R ₁ or C _1-3 halooxyalkyl optionally substituted by R ₁ ; R ₁ is selected from C _1-3 alkyl, hydroxyl, amino, nitro, cyano, oxo, carboxyl, carbonyl.

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

According to an embodiment of the present invention, R is selected from H, halogen or C _1-6 alkoxy.

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

According to an embodiment of the present invention, R is selected from C _1-6 alkoxy.

According to an embodiment of the present invention, R is selected from C _1-5 alkoxy.

According to an embodiment of the present invention, R is selected from C _1-4 alkoxy.

According to an embodiment of the present invention, R is selected from C _1-3 alkoxy.

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

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

Pharmaceutical composition

The present invention proposes a pharmaceutical composition, which includes the aforementioned compound, or its stereoisomers, tautomers, solvates, and pharmaceutically acceptable salts. As can be seen from the above, the aforementioned compound has SIRT6 agonist activity, and therefore, the pharmaceutical composition of the present invention can promote the expression or activity of SIRT6 protein, thereby effectively preventing and/or treating related diseases or symptoms mediated by SIRT6 protein.

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

The administration of the pharmaceutical composition of the present invention can be carried out by any acceptable mode of administration. The pharmaceutical composition of the present invention can be formulated into preparations 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 obvious 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 composition of the present invention is formulated so as to allow the biologically active ingredients contained therein to be bioavailable after the composition is administered to the patient.

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

In this article, the term "pharmaceutically acceptable excipient" may include any solvent, suitable for a particular target dosage form. Except for any conventional excipients that are incompatible with the compounds of the present disclosure, such as any adverse biological effects produced or interactions with any other components of the pharmaceutically acceptable composition in a harmful manner, their use is also contemplated by the present disclosure.

use

The present invention proposes the use of the aforementioned compounds as SIRT6 agonists. As can be seen from the foregoing, the aforementioned compounds have SIRT6 agonist activity, and using them as SIRT6 agonists can promote the expression or activity of SIRT6 protein, or effectively prevent and/or treat related diseases or symptoms mediated by SIRT6 protein.

The present invention proposes the use of the aforementioned compound in the preparation of a reagent for promoting the expression or activity of SIRT6 protein. As can be seen from the foregoing, the aforementioned compound has SIRT6 agonist activity, and using it as a reagent can effectively promote the expression or activity of SIRT6 protein, and can be particularly used to increase the expression or activity of SIRT6 protein in in vitro cells, for example, to construct a desired cell model.

The present invention proposes the use of the aforementioned compound or the aforementioned pharmaceutical composition in the preparation of a drug, and the drug is used to prevent and/or treat SIRT6 protein-mediated related diseases or symptoms. As can be seen from the above, the aforementioned compound has SIRT6 agonist activity, and using it as a SIRT6 drug can effectively prevent and/or treat SIRT6 protein-mediated related diseases or symptoms.

The present invention proposes that the aforementioned compound or the aforementioned pharmaceutical composition is used to prevent and/or treat SIRT6 protein-mediated related diseases or symptoms. As can be seen from the above, the aforementioned compound has SIRT6 agonist activity and can effectively prevent and/or treat SIRT6 protein-mediated related diseases or symptoms.

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.

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.

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 cancer.

In this article, "anti-aging" refers to the ability to prevent the aging of biological individuals, such as anti-skin aging, anti-cell aging, or delaying the shortening of telomere length.

method

The present invention proposes a method for activating and/or promoting the activity of SIRT6 protein in cells in vitro. According to an embodiment of the present invention, the method comprises: contacting the cells with the aforementioned compound. As can be seen from the foregoing, the aforementioned compound 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 cells.

The present invention proposes a method for preventing and/or treating related diseases or symptoms mediated by SIRT6 protein. According to an embodiment of the present invention, the method comprises: administering a pharmaceutically acceptable amount of the aforementioned compound or the aforementioned pharmaceutical composition to a subject. As can be seen from the above, the aforementioned compound has SIRT6 agonist activity. Thus, the above-mentioned compound can effectively prevent and/or treat related diseases or symptoms mediated by SIRT6 protein.

The effective amount of the compound of the present invention may vary depending on the mode of administration and the severity of the disease to be treated. The selection of the preferred effective amount can be determined by a person of ordinary skill in the art based on various factors (e.g., through clinical trials). The factors include, but are not limited to: pharmacokinetic parameters of the active ingredient such as bioavailability, metabolism, half-life, etc.; the severity of the disease to be treated, the patient's weight, the patient's immune status, the route of administration, etc. For example, several divided doses may be administered daily, or the dose may be reduced proportionally, depending on the urgency of the treatment situation.

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.

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.

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.

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 cancer.

The scheme of the present invention will be explained below in conjunction with the embodiments. It will be appreciated by those skilled in the art that the following embodiments are only used to illustrate the present invention and should not be considered as limiting the scope of the present invention. Where specific techniques or conditions are not indicated in the embodiments, the techniques or conditions described in the literature in this area or the product specifications are used. The reagents or instruments used are not indicated by the manufacturer and are all conventional products that can be obtained commercially.

Example 1: Acquisition of Compound 1

Compound 1 was synthesized by Shanghai TOPScience Technology Co., Ltd. The hydrogen spectrum of compound 1 is shown in FIG1 , and its hydrogen spectrum is characterized as follows:

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).

The structure of compound 1 is shown below:

Example 2: Activity Detection of Compound 1

1. Verification of SIRT6 enzyme activity of compound 1

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 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, and then the activation effect of the added compound on the SIRT6 enzyme activity is reflected. Among them, the detailed steps are as follows:

1) The in vitro enzyme activity of compound 1 in Example 1 was detected using a 384-well plate with a transparent bottom and black inner wall.

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

Table 1

Add the test compound with 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 system solution. The composition of the buffer system solution is shown in Table 2:

Table 2

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.

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

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

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.

2. Compound dose-effect curve detection

Take compound 1 in Example 1, set up different concentration gradients, and detect the compound at each concentration 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 show that compared with the existing SIRT6 enzyme activity agonist MDL-800, compound 1 has a better agonist effect in the in vitro enzyme activity detection system, and the log (IC50) of compound 1 is 1.988.

3. Compound toxicity testing

Compound 1 prepared in Example 1 was taken and configured into different concentration gradients, and then added to the culture supernatant of the mouse hippocampal neuron cell line HT22. After culturing at 37°C for 48 hours, CCK8 was used to detect the proliferation inhibition of cells at each concentration. The detection results of compound 1 are shown in Figure 4. The results show that compound 1 has strong cytotoxicity and can be used as a potential chemotherapeutic 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.

4. Detection of compound physiological functions

Compound 1 in Example 1 was taken, and different concentration gradients of Compound 1 were added to the culture supernatant of mouse hippocampal neuron cell line HT22. After constant temperature culture at 37°C for 48 hours, the protein of mouse hippocampal neuron cell line HT22 treated for 48 hours was taken, and the acetylation of H3K9 site was detected by Western blot, and the activation effect of Compound 1 on SIRT6 catalysis in cell lines was evaluated. The detection results of Compound 1 are shown in Figure 5. The results show that Compound 1 can effectively enhance the deacetylation activity of SIRT6 on histones in cell lines.

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

Considering that the cell line has the characteristics of immortalized cells and cannot well characterize the characteristics of senescent cells, the experiment was conducted in primary cells or C2C12 cell lines specifically used for studying senescence in this example.

At present, the common indicators for detecting the level of cell senescence in academia mainly include: SA-β-Gal staining and detection of the expression of p16 and p21 senescence-related genes. The specific steps are as follows:

1) β-galactose was used to induce senescence of C2C12 cell line, and compound 1 in Example 1 was added to the culture medium and co-cultured with the cells for 24 hours. The cells were collected to extract RNA, which was reverse transcribed into cDNA, and the expression of p16 and p21 genes in the cells was detected by fluorescent quantitative PCR.

2) β-galactose was used to induce senescence of the C2C12 cell line, and the compound 1 in 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 detect the effect of the addition of compound 1 on the positive rate of SA-β-Gal staining of the cells.

3) Inducing replicative senescence of primary cells by multiple passages, adding compound 1 in Example 1 to the culture medium of primary cells, and co-culturing with cells for 24 hours, collecting cells to extract RNA, reverse transcribe into cDNA, and using fluorescent quantitative PCR to detect the expression of p16 and p21 genes in cells.

4) The primary cells were induced to undergo replicative senescence by multiple passages, and the compound 1 in 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 detect the effect of the addition of the compound on the positive rate of SA-β-Gal staining of the cells.

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 the primary cells of induced senescent C2C12 that had produced replicative senescence and reached the division limit, which further demonstrated that the compounds of the present invention have anti-aging activity at the cellular level.

In the description of this specification, the description with reference to the terms "one embodiment", "some embodiments", "example", "specific example", or "some examples" etc. means 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 may be combined in any one or more embodiments or examples in a suitable manner. In addition, those skilled in the art may combine and combine the different embodiments or examples described in this specification and the features of the different embodiments or examples, without contradiction.

Although the embodiments of the present invention have been shown and described above, it is to be understood that the above embodiments are exemplary and are not to be construed as limitations of the present invention. A person skilled in the art may change, modify, replace and vary the above embodiments within the scope of the present invention.

Claims (13)

A class of compounds, which are compounds represented by formula (I) or stereoisomers, tautomers, solvates, and pharmaceutically acceptable salts thereof:
wherein ring A is selected from polyvalent aromatic rings; R is selected from -H, halogen, hydroxyl, amino, carbonyl, C _1-6 alkyl optionally substituted by R ₁ , C _{1-6 alkoxy optionally substituted by R 1, C 1-6 haloalkyl optionally} substituted by R 1, or C _1-6 halooxyalkyl optionally substituted by R _{1 ; R1} is selected from _C1-4 alkyl, hydroxy, amino, nitro, cyano, oxo, carboxyl, and carbonyl. 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. 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 groups. The compound according to claim 1, characterized in that the compound represented by formula (I) has the structure shown below:
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. The pharmaceutical composition according to claim 5, characterized in that the pharmaceutical composition further comprises a pharmaceutically acceptable carrier or excipient. 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. 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 drug for preventing and/or treating SIRT6 protein-mediated related diseases or symptoms. 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. The use according to claim 8 or 9 is characterized in that the related diseases or symptoms mediated by the SIRT6 protein include aging-related diseases, cancer, inflammation or anti-aging. The use according to claim 10 is 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. A method for activating and/or promoting the activity of cellular SIRT6 protein in vitro, comprising: The cell is contacted with the compound according to any one of claims 1 to 4. A method for preventing and/or treating SIRT6 protein-mediated diseases or symptoms, comprising: 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.