Small molecule compound having sulfonic acid or sulfinic acid lactone structure, composition and use thereof

Abstract

The present application discloses a small molecule compound having a sulfonic acid or sulfinic acid lactone structure, a composition and a use thereof. The present application provides a small molecule compound having a sulfonic acid or sulfinic acid lactone structure. It has been verified that the small molecule compound has extremely low skin irritation, excellent permeability and relatively high stability, and can not only significantly remove active oxygen and reduce oxidative stress, but also relieve skin inflammation. Thus, the small molecule compound has a good ability to protect and repair skin cells and can be used as a new raw material for cosmetics.

Description

Small molecule compound having sulfonic acid or sulfenic acid lactone structure, composition and application thereof

[0001] Cross-reference Statement

[0002] This patent application claims priority to the Chinese patent application filed on November 1, 2023, with application number 2023114494701 and invention name “Small molecule compounds, compositions and applications thereof having sulfonic acid or sulfinic acid lactone structure”. The full text of the above application is incorporated herein by reference. Technical Field

[0003] The present invention relates to the field of biochemistry, and in particular to a small molecule compound having a sulfonic acid or sulfenic acid lactone structure, a composition and applications thereof. Background Art

[0004] Protecting and repairing damaged skin is often a key function of skincare products and cosmetics. Factors that contribute to skin damage include hormone levels, excessive sun exposure, smog, and an improper diet. Inflammation and damage to the skin can lead to roughness, pigmentation, decreased collagen, and a decrease in the skin's antioxidant capacity, which can accelerate long-term skin aging. Studies have shown that quenching ROS and scavenging harmful oxygen free radicals can alleviate skin inflammation and repair damaged skin. While traditional antioxidants in existing cosmetics, such as vitamin C, vitamin E, niacinamide, and arbutin, possess some ROS scavenging capabilities, they still present significant irritation, low stability, and poor skin permeability.

[0005] Therefore, it is of great value to develop new cosmetic raw materials that are highly permeable, weakly irritating, and can quench ROS, scavenge harmful oxygen free radicals, and relieve skin inflammation.

[0006] Summary of the Invention

[0007] The object of the present invention is to provide a small molecule compound having a sulfonic acid or sulfenic acid lactone structure.

[0008] Another object of the present invention is to provide a composition.

[0009] Another object of the present invention is to provide a method for alleviating skin inflammation.

[0010] Another object of the present invention is to provide a method for reducing active oxygen in skin cells.

[0011] Another object of the present invention is to provide uses of the above-mentioned compound or composition.

[0012] To solve the above technical problems, the first aspect of the present invention provides a compound having a structure shown in the following general formula I, a salt, stereoisomer or solvate thereof,

[0013] Wherein, X is oxygen or none;

[0014] R 1 and R 2 are independently hydrogen, hydroxyl, C 1-6 Alkyl, C 1-6 Alkoxy, amino, -NC(O)R 1-1 、-NR a R b 、-CH2C(O)OR 1-2 , phenyl, -OC(O)R 1-3 、-OCH2OC(O)R 1-4 , halogen, cyano, nitro, -C(O)R 1-5 or-C(O)OR 1-6 ;

[0015] R 1-1 、R 1-2 、R 1-3 、R 1-4 、R 1-5 and R 1-6 are independently hydrogen, C 1-6 Alkyl, halogen-substituted C 1-6 Alkyl or -NR a R b ;

[0016] R a and R b are independently hydrogen, C 1-6 Alkyl or halogen substituted C 1-6 alkyl.

[0017] In some preferred embodiments, X is oxygen.

[0018] In some preferred embodiments, R 1 and R 2 are independently hydrogen, hydroxyl, C 1-6 Alkyl, C 1-6 Alkoxy, amino, -NC(O)R 1-1 、-NR a R b 、-CH2C(O)OR 1-2 , phenyl, -OC(O)R 1-3 or -OCH2OC(O)R 1-4 .

[0019] In some preferred embodiments, R 1 and R 2 are each independently hydroxyl, C 1-6 Alkoxy,

[0020] In some preferred embodiments, the C 1-6 Alkyl is C 1-4 More preferably, the C 1-4 The alkyl group is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl.

[0021] In some preferred embodiments, the C 1-6 Alkoxy is C 1-4 Alkoxy; more preferably, C 1-4 Alkoxy is methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy or tert-butoxy.

[0022] In some preferred embodiments, the halogen is fluorine, chlorine, bromine or iodine.

[0023] In some preferred embodiments, the halogen-substituted C 1-6 Alkyl is halogen substituted C 1-4 Alkyl; more preferably halogen-substituted methyl, halogen-substituted ethyl, halogen-substituted n-propyl, halogen-substituted isopropyl, halogen-substituted n-butyl, halogen-substituted isobutyl or halogen-substituted tert-butyl.

[0024] In some preferred embodiments, R 1 is hydroxy, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, More preferably, R 1 Hydroxyl, methoxy,

[0025] In some preferred embodiments, R 2 is hydroxy, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, More preferably, R 2 Hydroxyl, methoxy,

[0026] In some preferred embodiments, R 1-3 is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl or More preferably, R 1-3 isopropyl, tert-butyl or

[0027] In some preferred embodiments, R 1-4 is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl or More preferably, R 1-4 For tert-butyl.

[0028] In some preferred embodiments, the compound is selected from any one of the following:

[0029] In some preferred embodiments, the compound is not

[0030] The second aspect of the present invention provides a composition comprising the compound described in the first aspect of the present invention, a salt, stereoisomer or solvate thereof; and cosmetically acceptable excipients.

[0031] The third aspect of the present invention provides use of the compound described in the first aspect of the present invention, its salt, stereoisomer or solvate, or the composition described in the second aspect of the present invention in the preparation of cosmetics.

[0032] The fourth aspect of the present invention provides a method for alleviating skin inflammation, comprising the steps of: administering to a subject the compound, salt, stereoisomer or solvate thereof described in the first aspect of the present invention; or, administering to a subject the composition described in the second aspect of the present invention.

[0033] In a fifth aspect, the present invention provides a method for reducing reactive oxygen species in skin cells, the method comprising the steps of: administering to a subject the compound, salt, stereoisomer or solvate thereof described in the first aspect of the present invention; or administering to a subject the composition described in the second aspect of the present invention.

[0034] Compared with the prior art, the present invention has at least the following advantages:

[0035] The present invention provides a class of small molecule compounds with a sulfonic acid or sulfenic acid lactone structure, which have been proven to have extremely low skin irritation, excellent penetration ability and high stability. They can not only significantly scavenge reactive oxygen species and reduce oxidative stress, but also relieve skin inflammation. They have excellent ability to protect and repair skin cells and can be used as new cosmetic raw materials.

[0036] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features described in detail below (such as in the embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be listed here one by one. BRIEF DESCRIPTION OF THE DRAWINGS

[0037] One or more embodiments are exemplarily described by the figures in the corresponding drawings, and these exemplified descriptions do not constitute limitations on the embodiments.

[0038] FIG1 is a diagram showing the behavioral trajectory of a representative zebrafish selected from each group according to an embodiment of the present invention;

[0039] FIG2 is a statistical diagram of the total movement distance of each group of zebrafish within 20 minutes according to an embodiment of the present invention;

[0040] FIG3 is a statistical diagram of the average movement speed of each group of zebrafish within 20 minutes according to an embodiment of the present invention;

[0041] FIG4 is a representative image of inflammatory cell staining in zebrafish neuromasts of each group according to an embodiment of the present invention;

[0042] FIG5 is a statistical diagram of the number of inflammatory cells in each group of zebrafish neuromasts according to an embodiment of the present invention;

[0043] FIG6 shows the effect of I-8 on zebrafish wound repair according to an embodiment of the present invention;

[0044] FIG7 is a statistical diagram of the tail fin area of ​​zebrafish in each group according to an embodiment of the present invention;

[0045] FIG8 is a diagram showing the effect of I-8 on the number of inflammatory cells in zebrafish wounds according to an embodiment of the present invention;

[0046] FIG9 is a statistical diagram of the number of inflammatory cells in each group of wounds according to an embodiment of the present invention.

[0047] FIG10 shows the ROS content in zebrafish according to an embodiment of the present invention, wherein the white line area is the zebrafish yolk sac (ROS quantification area), and the values ​​in brackets are I-8 concentrations in μg / mL;

[0048] FIG11 is a statistical diagram of ROS levels in zebrafish according to an embodiment of the present invention;

[0049] FIG12 is a picture of β-galactosidase activity in zebrafish according to an embodiment of the present invention. After β-galactosidase staining, the tissue appears blue-green. Comparing the tissues pointed by arrows on the dorsal fin, the more arrows there are, the darker the blue-green color, and the higher the β-galactosidase activity.

[0050] FIG13 is a statistical diagram of β-galactosidase activity in zebrafish according to an embodiment of the present invention;

[0051] FIG14 is a statistical diagram of the expression of genes related to antioxidant and anti-aging in zebrafish according to an embodiment of the present invention;

[0052] Figure 15 shows that compound I-8 according to an embodiment of the present invention can down-regulate TNF-α in HaCaT;

[0053] FIG16 shows that compound I-8 according to an embodiment of the present invention can down-regulate IL-8 in HaCaT;

[0054] FIG17 shows that compound I-8 according to an embodiment of the present invention can down-regulate IL-1β in HaCaT;

[0055] FIG18 shows that compound I-8 according to an embodiment of the present invention can down-regulate EP2 of HaCaT;

[0056] FIG19 shows that compound I-8 according to an embodiment of the present invention can downregulate TRPV1 of HaCaT;

[0057] Figure 20 shows the scavenging of ROS by compound I-8 according to an embodiment of the present invention;

[0058] FIG21 shows that compound I-8 improves the secretion of type I collagen according to an embodiment of the present invention;

[0059] Figure 22 shows that compound I-8 up-regulates COL3A1 gene expression according to an embodiment of the present invention;

[0060] Figure 23 shows that compound I-8 up-regulates COL4A1 gene expression according to an embodiment of the present invention;

[0061] Figure 24 shows that compound I-8 up-regulates COL7A1 gene expression according to an embodiment of the present invention;

[0062] FIG25 shows that compound I-8 up-regulates LAMA5 gene expression according to an example of the present invention. DETAILED DESCRIPTION

[0063] Due to the poor stability of traditional cosmetic antioxidants, the application conditions are demanding, such as being decomposed by light during the day and can only be applied at night; poor skin permeability often requires high concentrations and large amounts of application to be effective, and some individuals often have no effect; in addition, these antioxidants also have strong irritation, and some fragile skins need to strictly control the dosage, otherwise they are prone to cause severe allergic and inflammatory reactions. The inventors have developed a class of small molecule polyphenol compounds with sulfonic acid or sulfenic acid lactone structures through extensive and in-depth research. These compounds have low irritation, good permeability, high stability, and can significantly quench reactive oxygen species (ROS), remove harmful oxygen free radicals, eliminate oxidative stress, alleviate skin inflammation, and soothe and repair damaged skin cells. Such compounds and their salts, stereoisomers or solvates can be used as new raw materials for cosmetics and have the potential to replace existing antioxidants.

[0064] Compound

[0065] The present invention relates to a class of small molecule compounds having a sulfonic acid or sulfenic acid lactone structure, the structure of which is shown in the following general formula I:

[0066] In the above general formula I, X is oxygen or none. Based on the beneficial effect of further improving the stability and permeability of the compound, X is oxygen.

[0067] R 1 and R 2

[0068] R 1 The number of is 0-4, such as 0, 1, 2, 3 or 4. 1 , can be hydrogen, hydroxyl, C 1-6 Alkyl, C 1-6 Alkoxy, amino, -NC(O)R 1-1 、-NR a R b 、-CH2C(O)OR 1-2 , phenyl, -OC(O)R 1-3 、-OCH2OC(O)R 1-4 , halogen, cyano, nitro, -C(O)R 1-5 or -COOR 1-6 ; and, R 1-1 、R 1-2 、R 1-3 、R 1-4 、R 1-5 and R 1-6 are independently hydrogen, C 1-6 Alkyl, halogen-substituted C 1-6 Alkyl or -NR a R b ; R a and R b are independently hydrogen, C 1-6 Alkyl or halogen substituted C 1-6 In a preferred embodiment, R 1 For hydrogen, hydroxyl, C 1-6 Alkyl, C 1-6 Alkoxy, amino, -NC(O)R 1-1 、-NR a R b 、-CH2C(O)OR 1-2 , phenyl, -OC(O)R 1-3 or -OCH2OC(O)R 1-4 .

[0069] Likewise, R 2 The number of is 0-4, such as 0, 1, 2, 3 or 4. 2 , can be hydrogen, hydroxyl, C 1-6 Alkyl, C 1-6 Alkoxy, amino, -NC(O)R1-1 、-NR a R b 、-CH2C(O)OR 1-2 , phenyl, -OC(O)R 1-3 、-OCH2OC(O)R 1-4 , halogen, cyano, nitro, -C(O)R 1-5 or -COOR 1-6 ; and, R 1-1 、R 1-2 、R 1-3 、R 1-4 、R 1-5 and R 1-6 are independently hydrogen, C 1-6 Alkyl, halogen-substituted C 1-6 Alkyl or -NR a R b ; R a and R b are independently hydrogen, C 1-6 Alkyl or halogen substituted C 1-6 In a preferred embodiment, R 2 For hydrogen, hydroxyl, C 1-6 Alkyl, C 1-6 Alkoxy, amino, -NC(O)R 1-1 、-NR a R b 、-CH2C(O)OR 1-2 , phenyl, -OC(O)R 1-3 or -OCH2OC(O)R 1-4 .

[0070] In a more preferred embodiment of the present invention, the above-mentioned R 1-3 is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl or More preferably, R 1-3 isopropyl, tert-butyl or R 1-4 is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl or More preferably, R 1-4 For tert-butyl.

[0071] R 1 and R 2 The numbers of R 1 and R 2 The number of is equal.

[0072] R 1 and R 2The substitution position is not limited, but in a preferred embodiment, R 1 and R 2 The positions of the biphenyl ring are respectively 4 and 4' (as shown in I-8 in a specific embodiment of the present invention). As used in the present invention, the position numbering rule of the biphenyl ring is: In the sulfonic acid or sulfinic acid lactone structure, the O atom is covalently bonded to the carbon on 2', the S atom is covalently bonded to the carbon on 2, and the oxygen atom and the sulfur atom are covalently bonded to each other to form a lactone structure.

[0073] Based on the excellent effect of promoting the compound to clear active oxygen and alleviate verification, in a more preferred embodiment, R 1 and R 2 At least one of them has the ability to donate electrons to the parent ring, and preferably both have the ability to donate electrons to the target. 1 and R 2 are each independently hydroxyl, C 1-6 Alkoxy, More preferably, R 1 and R 2 is hydroxy, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, In a specific embodiment of the present invention, R 1 Hydroxyl, methoxy, In a specific embodiment of the present invention, R 2 Hydroxyl, methoxy,

[0074] In certain embodiments of the present invention, the specific structures of the compounds are shown in the following table, wherein compounds I-1 to I-12, I-17 and I-18 have better penetration ability than I-13 to I-16.

[0075] surface

[0076] Composition

[0077] The present invention also relates to a composition containing the compound of the present invention, its salt, stereoisomer or solvate. The composition of the present invention may be a cosmetic composition or an external preparation.

[0078] As used in the present invention, the term "cosmetics" refers to products that are applied to the human body surface (such as the epidermis, hair, lips, etc.) by smearing, spraying or other similar methods to clean, maintain, beautify, or eliminate bad odors, and the product has a soothing effect on the application area.

[0079] When used as a cosmetic composition, the compound of the present invention, its salt, stereoisomer or solvate composition is used as a cosmetic benefit agent in the cosmetic composition. In addition, the composition also includes a cosmetically acceptable medium for diluting, dispersing or serving as a carrier for the cosmetic benefit agent to promote its distribution when the composition is applied to the skin.

[0080] These media can be aqueous, anhydrous or emulsions. Oily carriers, in the presence of water and an emulsifier, can form an emulsion system as a carrier. Preferably, the composition is aqueous or an emulsion, especially a water-in-oil or oil-in-water emulsion, preferably an oil-in-water emulsion. Cosmetic compositions are generally in the form of, but not limited to, liquids, creams or emulsions. In a preferred embodiment, water is used as a carrier to form the cosmetic composition. In a preferred embodiment, the cosmetic composition further comprises a carrier other than water, such as oil, fat, wax oily base (such as coconut oil, palm oil, olive oil, castor oil, mink oil, snake oil, silicone oil and its derivatives, tallow, lanolin and its derivatives, carnauba wax, spermaceti, beeswax, liquid paraffin, vaseline, microcrystalline wax, squalane, fatty acids, fatty alcohols and esters, etc.), powdery base (such as talc, kaolin, zinc white, titanium dioxide, bentonite, magnesium stearate, zinc stearate, calcium carbonate, magnesium carbonate, calcium hydrogen phosphate, etc.), and solvent base (such as alcohols, small molecule ketones, ethers, small molecule esters).

[0081] In the cosmetic composition of the present invention, cosmetically acceptable excipients may also be added. The term "cosmetically acceptable excipients" refers to substances that play a role in the formation, stability, or imparting of color, fragrance, and other characteristics of cosmetics. For example, preservatives (such as benzoic acid and its derivatives, chlorobutanol, chloroxylenol, salicylic acid and its derivatives, sorbic acid and its derivatives, imidazolidinyl urea, phenylethanol, etc.), antioxidants (such as butylated hydroxyanisole, tert-butylated hydroxyanisole, vitamin E, propyl gallate, etc.), moisturizers (such as glycerin, propylene glycol, sorbitol, polyethylene glycol, lactic acid, sodium lactate, sodium pyrrolidonecarboxylate, hyaluronic acid, hydrolyzed collagen, chitin and its derivatives, glucose esters), sunscreens (titanium dioxide, zinc oxide, aminobenzoic acid esters and their derivatives, salicylic acid and its derivatives, benzophenone, etc.), surfactants (lecithin, soap beans), glycosides, alkyl glycosides, etc.), antioxidants, colorants (such as organic synthetic pigments azo series, anthraquinone series, etc., or inorganic pigments such as zinc oxide, titanium dioxide, ferric oxide, ferrous hydroxide, chromium trioxide, ferric oxide, etc., or natural pigments capsaicin, sorghum red, anthocyanins, etc.), flavors (natural or blended flavors such as ambergris, castoreum, musk, civet, rose oil, peppermint oil, spearmint oil, lavender oil, fennel oil, etc.), water-soluble polymers, chelating agents (such as disodium edetate and its derivatives) and film agents (such as polyvinyl acetate, polyacrylate emulsion, hyaluronic acid, polyglyceryl-2 isostearate, etc.), etc.

[0082] In the cosmetic composition of the present invention, there is no restriction on the amount of each raw material and auxiliary material added.

[0083] Uses of compounds or compositions

[0084] The present invention also relates to the use of the above-mentioned compound, its salt, stereoisomer or solvate, or a composition containing the above-mentioned compound, its salt, stereoisomer or solvate, for: (i) preparing cosmetics; (ii) relieving skin inflammation; (iii) reducing active oxygen in skin cells; and / or (iv) repairing damaged skin.

[0085] As used herein, the term "skin" includes the skin on the face, neck, chest, back, arms, axillae, arms, hands, legs, and scalp. As used herein, a cosmetic benefit agent is meant to include a component that (a) improves a facial or body feature, such as a skin feature, upon topical application, (b) benefits a facial or body feature, such as a skin feature, or (c) both (a) and (b). In preferred embodiments, the compound or cosmetic composition is for topical application.

[0086] Method for relieving skin inflammation and / or reducing active oxygen in skin cells and / or repairing damaged skin

[0087] The present invention also relates to a method for (a) alleviating skin inflammation and / or (b) reducing reactive oxygen species in skin cells and / or (c) repairing damaged skin, comprising the steps of administering the aforementioned compound, salt, stereoisomer, or solvate thereof to a subject; or administering the aforementioned composition to a subject. The administration is parenteral, preferably topical, such as by smearing or applying to the skin, mucous membranes, or the like.

[0088] the term

[0089] As used herein, the term "alkyl" refers to a linear or branched saturated monovalent hydrocarbon group, wherein the alkyl group may be optionally substituted with one or more substituents. In a specific embodiment, the alkyl group is a group having 1 to 20 (C 1-20 ), 1 to 15 (C 1-15 ), 1 to 12 (C 1-12 ), 1 to 10 (C 1-10 ) or 1 to 6 (C 1-6 ) carbon atoms, or a linear saturated monovalent hydrocarbon group having 3 to 20 (C 3-20 ), 3 to 15 (C 3-15 ), 3 to 12 (C 3-12 ), 3 to 10 (C 3-10 ) or 3 to 6 (C 3-6 ) carbon atoms with a branched saturated monovalent hydrocarbon group. The linear C 1-6 and branched C 3-6 Alkyl groups are also referred to as "lower alkyl groups". Examples of alkyl groups include, but are not limited to, methyl, ethyl, propyl (including all isomeric forms), n-propyl, isopropyl, butyl (including all isomeric forms), n-butyl, isobutyl, tert-butyl, pentyl (including all isomeric forms) and hexyl (including all isomeric forms). For example, C 1-6 Alkyl refers to a linear saturated monovalent hydrocarbon radical having 1 to 6 carbon atoms or a branched saturated monovalent hydrocarbon radical having 3 to 6 carbon atoms. In one embodiment, the alkyl radical is an optionally substituted alkyl radical as described elsewhere herein. In some embodiments, C 1-6 Alkyl is C 1-4 Alkyl, C 1-4 Alkyl is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl. 1-6 In other embodiments, any one or more hydrogen atoms in the alkyl group are replaced by halogen atoms. 1-6 Alkyl is halogen substituted C 1-4 In other embodiments, the halogen-substituted C 1-4The alkyl group is a halogen-substituted methyl group, a halogen-substituted ethyl group, a halogen-substituted n-propyl group, a halogen-substituted isopropyl group, a halogen-substituted n-butyl group, a halogen-substituted isobutyl group or a halogen-substituted tert-butyl group.

[0090] As used herein, the term "alkoxy" refers to a stable linear or branched, or cyclic hydrocarbon group, or a combination thereof, consisting of the indicated number of carbon atoms and one or more (in one embodiment, one to three) O atoms. Examples of alkoxy groups include, but are not limited to -O-CH3, -O-CH2-CH3, -O-CH2-CH2-CH3, -O-CH-(CH3)2, and -O-CH2-CH2-O-CH3. In one embodiment, the alkoxy group is an optionally substituted alkoxy group described elsewhere herein. In some embodiments, the alkoxy group is C 1-6 In some embodiments, the alkoxy group is C 1-4 In some embodiments, the alkoxy group is methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy or tert-butoxy. In other embodiments, C 1-6 Any one or more hydrogen atoms in the alkoxy group are replaced by halogen atoms. 1-6 The alkoxy group is a halogen-substituted methoxy group, a halogen-substituted ethoxy group, a halogen-substituted n-propoxy group, a halogen-substituted isopropoxy group, a halogen-substituted n-butoxy group, a halogen-substituted isobutoxy group or a halogen-substituted tert-butoxy group.

[0091] As used herein, the term "halogen" refers to fluorine, chlorine, bromine or iodine.

[0092] As used in the present invention, "(O)" is a =O structure, such as -NH-C(O)- is an amide group, -OC(O)- is an acyloxy group, and -C(O)- is a carbonyl group.

[0093] As used herein, the term "hydrogen" includes protons (1H), deuterium (2H), tritium (3H) and / or mixtures thereof. In the compounds described herein, one or more positions occupied by hydrogen may be enriched with deuterium and / or tritium. Such isotopically enriched analogs can be prepared by appropriately isotopically labeled starting materials obtained from commercial sources or by known literature procedures.

[0094] As used herein, the term "hydroxy" refers to -OH.

[0095] As used herein, the term "amino" refers to -NH2.

[0096] As used herein, the term "cyano" refers to -CN.

[0097] As used herein, the term "nitro" refers to -NO2.

[0098] The term "substituted" refers to moieties having substituents replacing a hydrogen or one or more non-hydrogen atoms on the molecule.

[0099] As used herein, the term "solvate" refers to a compound formed by the interaction of a solvent with a compound provided herein or a salt thereof.

[0100] In order to make the purpose, technical solutions and advantages of the embodiments of the present invention clearer, the present invention will be further described below in conjunction with specific examples. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention. The experimental methods in the following examples, for which specific conditions are not specified, are usually based on conventional conditions or the conditions recommended by the manufacturer. Unless otherwise stated, percentages and parts are weight percentages and parts by weight. The experimental materials and reagents used in the following examples can be obtained from commercial sources unless otherwise specified.

[0101] Unless otherwise specified, the technical and scientific terms used herein have the same meaning as commonly understood by ordinary technicians in the technical field to which this application belongs. It should be noted that the terms used herein are only for describing specific embodiments and are not intended to limit the exemplary embodiments of this application.

[0102] General synthetic step 1

[0103] General Synthesis Step 2

[0104] General Synthesis Step 3

[0105] General Synthesis Step 4

[0106] Example 1: Synthesis and characterization of compound I-8

[0107] Synthesis of Intermediate 2: 2-Iodo-5-methoxyphenol (0.73 g, 2.92 mmol, 1.2 eq) and DMAP (0.03 g, 0.24 mmol, 0.1 eq) were dissolved in DCM (5.00 mL) and pyridine (10.00 mL). The mixture was cooled to 0°C and stirred until uniform. Compound 1 (0.50 g, 2.43 mmol, 1 eq) was then dissolved in DCM (5.00 mL). This solution was then slowly added dropwise to the reaction system, maintained at 0°C. After the addition was complete, the system was allowed to warm to room temperature and allowed to react overnight. LC-MS indicated complete reaction of the starting material. Stirring was stopped, and the mixture was extracted with water and DCM. The organic phase was separated, concentrated, and purified on a silica gel column (PE:EA = 10:1) to afford Intermediate 2 (0.96 g, 2.23 mmol, 94.61% yield, 93.42% purity) as a yellow oil. LC-MS: [M+1] + =421.

[0108] Synthesis of intermediate 3 (i.e., I-17): Compound 2 (0.2 g, 0.48 mmol, 1 eq), Pd(Pivate)2 (0.015 g, 0.048 mmol, 0.1 eq), TBAB (0.18 g, 0.57 mmol, 1.2 eq), and potassium acetate (0.14 g, 1.43 mmol, 3 eq) were dissolved in DMAc (2.00 mL), fully replaced with nitrogen, and the temperature was raised to 60 ° C. The reaction was stirred under nitrogen protection overnight. The reaction was completed by LC-MS monitoring, filtered, extracted with EA, concentrated, and purified on a silica gel column (PE: EA = 1: 1) to give intermediate 3 (0.12 g, 0.42 mmol, 88.49% yield, 93.32% purity) as a white solid. 1 H NMR(400MHz,Chloroform-d)δ7.76(d,J=2.8Hz,1H),7.74(d,J=2.7Hz,1H),7.43(d,J=2.7Hz,1H),7.28– 7.23(m,1H),6.93(dd,J=8.8,2.6Hz,1H),6.84(d,J=2.6Hz,1H),3.91(s,3H),3.87(s,3H).LC-MS: [M+1] + =293.

[0109] Synthesis of compound I-8: Compound 3 (1.00 g, 3.42 mmol, 1 eq) was dissolved in DCM (20.00 mL), cooled to 0°C, and BBr3 (1.0 M in DCM, 13.68 mmol, 13.68 mL, 4 eq) was slowly added dropwise under nitrogen protection. The temperature was controlled at 0°C. After the addition was complete, the system was allowed to naturally warm to room temperature and reacted overnight. The reaction was monitored by LC-MS. The reaction was quenched with ice water, extracted with EA and DCM, concentrated, and purified by silica gel column (PE: EA = 1: 1) to obtain compound I-8 (0.25 g, 0.95 mmol, 27.78% yield, 100% purity) as a light yellow solid. 1H NMR (400MHz, DMSO-d6) δ7.96(d,J=8.5Hz,1H),7.92(d,J=8.8Hz,1H),7.27–7.22(m,2H),6.87(dd,J=8.6,2.4Hz,1H),6.78(d,J=2.4Hz,1H).LC-MS: [M-1] - =263.

[0110] Example 2: Synthesis and Characterization of Compound I-11

[0111] In Example 1, the reaction of synthesizing compound 3 from compound 2 also produces compound 3'. Compound 3' can be subjected to the same demethylation step to obtain compound I-11. LC-MS: [M-1] - =263.

[0112] Example 3: Synthesis and characterization of compound I-13

[0113] Compound I-8 (130 mg, 0.49 mmol), DMAP (6 mg, 0.05 mmol) and DIEA (254 mg, 1.97 mmol) were dissolved in THF (5 mL), cooled to 0°C, and then dimethylaminoformyl chloride (212 mg, 1.97 mmol) was slowly added dropwise to the reaction system. The temperature was controlled at 0°C. After the addition was completed, the system was allowed to warm to room temperature naturally. The reaction was allowed to proceed overnight, concentrated, and purified on a silica gel column (DCM:EA=100:5) to obtain I-13 (90 mg, yield 45.01%) as a white solid. 1H NMR (400MHz, DMSO-d6) δ8.33(d,J=8.8Hz,1H),8.30(d,J=8.8Hz,1H),7.92(d,J=2.4Hz,1H),7.74(dd,J=8.7,2.5Hz,1H) ,7.45(d,J=2.3Hz,1H),7.34(dd,J=8.7,2.4Hz,1H),3.08(d,J=6.2Hz,6H),2.94(d,J=2.8Hz,6H).LCMS:m / z=407.1(M+H + ,ESI).

[0114] Example 4: Synthesis and Characterization of Compounds I-14 and I-15

[0115] Using the synthesis method in Example 3, I-8 was used as the starting material, and dimethylaminoformyl chloride was replaced with isobutyryl chloride or pivaloyl chloride to obtain I-14 and I-15, both of which were white solids. I-14: LCMS: m / z=405 (M+H + ,ESI).I-15: 1 H NMR (400MHz, DMSO-d6) δ8.24(d,J=8.8Hz,1H),8.07(dd,J=7.7,1.3Hz,1H),7.83(t,J=8.0Hz,1H),7.75(dd,J=8 .3,1.3Hz,1H),7.54(d,J=2.4Hz,1H),7.38(dd,J=8.8,2.4Hz,1H),1.35(s,9H),1.32(s,9H).LCMS:m / z=433(M+H + ,ESI).

[0116] Example 5: Synthesis and Characterization of Compound I-1

[0117] According to the general synthesis steps 1 and 2, the synthesis method in Example 1 was used to prepare compound As raw materials, compound I-1 was prepared: LCMS: m / z=247 (MH + ,ESI).

[0118] Example 6: Synthesis and Characterization of Compound I-2

[0119] According to the general synthesis steps 1 and 2, the synthesis method in Example 1 was used to prepare compound As raw materials, compound I-2 was prepared: LCMS: m / z=295 (MH + ,ESI).

[0120] Example 7: Synthesis and Characterization of Compound I-3

[0121] According to the general synthesis steps 1 and 2, the synthesis method in Example 1 was used to prepare compound As raw materials, compound I-3 was prepared: LCMS: m / z=247 (MH + ,ESI).

[0122] Example 8: Synthesis and Characterization of Compound I-4

[0123] According to the general synthesis steps 1 and 2, the synthesis method in Example 1 was used to prepare compound As raw materials, compound I-1 was prepared: LCMS: m / z=311 (MH + ,ESI).

[0124] Example 9: Synthesis and Characterization of Compound I-5

[0125] According to the general synthesis steps 1 and 2, the synthesis method in Example 1 was used to prepare compound As raw materials, compound I-5 was prepared: LCMS: m / z=279 (MH + ,ESI).

[0126] Example 10: Synthesis and Characterization of Compound I-6

[0127] According to the general synthesis steps 1 and 2, the synthesis method in Example 1 was used to prepare compound As raw materials, compound I-6 was prepared: LCMS: m / z=295 (MH + ,ESI).

[0128] Example 11: Synthesis and Characterization of Compound I-7

[0129] According to the general synthesis steps 1 and 2, the synthesis method in Example 1 was used to prepare compound As raw materials, compound I-7 was prepared: LCMS: m / z=295 (MH + ,ESI).

[0130] Example 12: Synthesis and Characterization of Compound I-9

[0131] According to the general synthesis steps 1 and 2, the synthesis method in Example 1 was used to prepare compound As raw materials, compound I-9 was prepared: LCMS: m / z=263 (MH+ ,ESI).

[0132] Example 13: Synthesis and Characterization of Compound I-10

[0133] According to the general synthesis steps 1 and 2, the synthesis method in Example 1 was used to prepare compound As raw materials, compound I-10 was prepared: LCMS: m / z=279 (MH + ,ESI).

[0134] Example 14: Synthesis and Characterization of Compound I-12

[0135] According to the general synthesis steps 1 and 2, the synthesis method in Example 1 was used to prepare compound As raw materials, compound I-12 was prepared: LCMS: m / z=247 (MH + ,ESI).

[0136] Example 15: Synthesis and Characterization of Compound I-16

[0137] According to the general synthesis step 4, compound I-3 and As raw materials, compound I-16 was prepared: LCMS: m / z=493 (M+H + ,ESI).

[0138] Example 16: Synthesis and Characterization of Compound I-18

[0139] According to the general synthetic step 1, compound As raw materials, intermediate 4 was prepared:

[0140] Intermediate 4 was hydrogenated to remove the benzyl protection to prepare compound I-18: LCMS: m / z=293 (MH + ,ESI).

[0141] Example 17: Synthesis and Characterization of Compound I-19

[0142] According to the general synthetic step 1, compound As raw materials, compound I-19 was prepared: 1HNMR (400MHz, DMSO-d6) δ10.95(s,1H),10.67(s,1H),8.58(d,J=8.9Hz,1H),7.32–7.29(m,1H),7.28–7. 26(m,1H),7.26–7.23(m,1H),6.98(dd,J=8.4,1.2Hz,1H),6.90(dd,J=8.1,1.2Hz,1H).LCMS:m / z=263(MH + ,ESI).

[0143] Example 18: Synthesis and Characterization of Compound I-20

[0144] According to the general synthetic step 1, compound As raw materials, compound I-20 was prepared: 1 H-NMR(400MHz,DMSO-d6)δ10.77(s,1H),10.46(s,1H),10.22(s,1H),8.45–8.43(d,1H),7.2 5-7.24(d,1H),7.21–7.18(dd,1H),6.45–6.44(d,1H),6.28-6.27(d,1H).LCMS: m / z=279(MH + ,ESI).

[0145] Example 19: Synthesis and Characterization of Compound I-21

[0146] According to the general synthetic step 1, compound As raw materials, intermediate 5 was prepared:

[0147] Intermediate 5 was hydrogenated to remove the benzyl protection to prepare compound I-21: 1 H-NMR (400MHz, DMSO-d6) δ10.48(s,1H),8.07(d,J=8.7Hz,1H),7.99(d,J=8.7Hz,1H),7.48–7 .41(m,2H),6.89(dd,J=8.7,2.4Hz,1H),6.81(d,J=2.4Hz,1H),3.90(s,3H).LCMS:m / z=277(MH + ,ESI).

[0148] Example 20: Synthesis and Characterization of Compound I-22

[0149] According to the general synthetic step 1, compound As raw materials, intermediate 6 was prepared:

[0150] Intermediate 6 was hydrogenated to remove the benzyl protection to prepare compound I-22: 1 H-NMR (400MHz, DMSO-d6) δ11.04(s,1H),8.67(d,J=9.0Hz,1H),7.49(d,J=2.8Hz,1H),7.44(dd,J=9.1,2.9Hz,1H ),7.30(t,J=8.2Hz,1H),7.01(dd,J=8.3,1.2Hz,1H),6.92(dd,J=8.1,1.1Hz,1H),3.91(s,3H).LCMS:m / z=277(MH + ,ESI).

[0151] Example 21: Synthesis and Characterization of Compound I-23

[0152] According to the general synthetic step 1, compound As raw materials, compound I-23 was prepared: LCMS: m / z=277 (MH + ,ESI).

[0153] Example 22: Synthesis and Characterization of Compound I-24

[0154] According to the general synthetic step 1, compound As raw materials, compound I-24 was prepared: LCMS: m / z=281 (MH + ,ESI).

[0155] Example 23: Synthesis and Characterization of Compound I-25

[0156] According to the general synthetic step 1, compound As raw materials, compound I-25 was prepared: LCMS: m / z=297 (MH + ,ESI).

[0157] Example 24: Synthesis and Characterization of Compound I-26

[0158] According to the general synthetic step 1, compound As raw materials, compound I-26 was prepared: LCMS: m / z=277 (MH + ,ESI).

[0159] Example 25: Synthesis and Characterization of Compound I-27

[0160] According to the general synthetic step 1, compound As raw materials, compound I-27 was prepared: LCMS: m / z=281 (MH +,ESI).

[0161] Example 26: Synthesis and Characterization of Compound I-28

[0162] According to the general synthetic step 1, compound As raw materials, compound I-28 was prepared: LCMS: m / z=297 (MH + ,ESI).

[0163] Test Example 1: Zebrafish model to evaluate the efficacy of compounds in relieving stinging pain and relieving inflammation

[0164] Experimental animals: healthy wild AB zebrafish at 3 dpf (days post fertilization)

[0165] Evaluation of the pain-relieving efficacy: Healthy AB zebrafish were randomly divided into blank control, modeling, positive control, and sample groups. The blank control group was treated with embryo culture water; the modeling group was treated with the modeling drug (100μM SDS, sodium dodecyl sulfate); the positive control group was treated with both the modeling drug and the positive drug (200μM asiaticoside solution); and the sample group was treated with both the modeling drug and the test sample at different concentrations (high, medium, and low). After a period of continuous treatment, the zebrafish's behavioral trajectories were recorded using a behavioral analyzer, and changes in their total movement distance and speed were statistically analyzed. The results are presented in Figures 2-4. SDS causes stinging pain in the zebrafish, prompting them to move more vigorously. Consequently, the total movement distance and average speed of the modeling group were significantly increased compared to the blank control group. In the behavioral trajectory diagram in Figure 2, the black lines represent the zebrafish's movement within the well. More and more chaotic black lines indicate more vigorous movement.

[0166] Evaluation of soothing and anti-inflammatory efficacy: As described above, the modeling agent was 40 μM copper sulfate, the positive agent was 200 μM asiaticoside, and the I-8 concentrations were 66.6, 22.2, and 7.4 μg / mL. After zebrafish were treated as described above, inflammatory cells were stained, the number of inflammatory cells in the neuromasts was counted, and statistical analysis was performed. The results are presented in Figures 5-6.

[0167] Conclusion: I-8 has a certain effect in relieving stinging pain at the concentrations of 22.2, 66.6, and 200 μg / mL. It also has a certain effect in relieving inflammation at the concentrations of 7.4, 22.2, and 66.6 μg / mL.

[0168] Test Example 2: Evaluation of the ability of compounds to penetrate the skin using a Franz diffusion cell

[0169] Experimental materials: one-month-old Bama Xiang pig back skin, thickness 0.8-1.0 mm; 0.1 mg / mL test compound solution, solvent 50 mM Sodium Phosphate Buffer (PB) containing 20% ​​ethanol.

[0170] Experimental process:

[0171] 1. Pipette 400uL of the test compound solution and add it to the supply pool. Repeat three times.

[0172] 2. Add approximately 7600uL of 50mM PB buffer solution to the corresponding receiving cell

[0173] 3. Permeate the test compound at 32°C for 2h and 6h

[0174] 4. At 2h and 6h, the skin was washed with 50mM PB buffer solution containing 20% ​​ethanol, the skin of the infiltrated area was removed and homogenized, and acetonitrile solution containing internal standard was added.

[0175] 5. Centrifuge, take 100uL of the supernatant and add it to 100uL of pure water, mix well and use LC-MS / MS for quantitative analysis

[0176] The quantitative test results of the example compounds penetrating into the skin are shown in Table 2 below:

[0177] Table 2

[0178] Compound I-8 has good skin penetration ability and can reach a penetration amount of 10ug / g within 6 hours.

[0179] Test Example 3: Zebrafish Model to Evaluate the Repair Efficacy of Compounds

[0180] Experimental animals: healthy wild AB zebrafish at 3 dpf (days post fertilization)

[0181] Evaluation of wound repair efficacy: Healthy AB zebrafish at 3 dpf were used as experimental animals and randomly divided into a blank control group, a model group, a positive control group, and a sample group. The zebrafish in the blank control group were treated with culture water; the zebrafish in the model group were treated with culture water after the tail fin was cut; the zebrafish in the positive control and sample groups were treated with 600 μg / mL hyaluronic acid and different concentrations of I-8 (1, 3, 9 μg / mL) after the tail fin was cut. The fry in each group were placed in a constant temperature incubator at 28.5°C and kept away from light for 48 hours, with the medium changed every 24 hours. Image-Pro Plus software was used to calculate the tail fin area of ​​each group of zebrafish, and GraphPad software was used to perform statistical analysis of the data. The results are presented in Figures 7 and 8.

[0182] Evaluation of the restorative and anti-inflammatory efficacy: As described above, each group of larvae was placed in a constant-temperature incubator at 28.5°C, shielded from light, for 24 hours. Inflammatory cells in the fish were labeled with neutral red, and the number of inflammatory cells at the tail site was counted. GraphPad software was used for statistical analysis. The results are presented in Figures 9 and 10.

[0183] Conclusion: As shown in Figures 7 and 8, compared with the blank control group, the zebrafish tail fin regenerated in the model group, but its area was significantly reduced. When I-8 was applied to the zebrafish tail amputation model at concentrations of 1, 3, and 9 μg / mL, the tail fin area increased compared to the model group, with statistical significance. These results suggest that I-8 at concentrations of 1, 3, and 9 μg / mL can further promote the regeneration of tail fin tissue and have a certain effect on wound repair. As shown in Figures 9 and 10, compared with the blank control group, the number of inflammatory cells at the tail amputation site in the model group increased significantly. When I-8 was applied to the zebrafish tail amputation model at concentrations of 1, 3, and 9 μg / mL, the number of inflammatory cells at the wound site decreased compared to the model group, with statistical significance. These results suggest that I-8 at concentrations of 1, 3, and 9 μg / mL has a certain effect on inflammatory repair.

[0184] Test Example 4: Comparison of Kinetic Solubility in Aqueous Buffer with Urolithin A

[0185] Experimental Method: 30 μL of a 10 mM DMSO stock solution of the test compound was placed in duplicate in a solubility plate. 970 μL of aqueous buffer was then added to the plate. The plate was sealed with a film sealer and shaken at 1100 rpm for 2 h at 25°C. The sample from the solubility plate was transferred to a filter plate and filtered to obtain the filtrate. 10 μL of the filtrate was added along with 10 μL of DMSO to 980 μL of methanol, followed by a 10-fold dilution with methanol:water (1:1). The resulting sample was analyzed by LC-MS / MS to obtain the Area (filtered). Separately, 10 mM of the DMSO stock solution of the test compound was diluted to 300 μM with DMSO. 10 μL of this 300 μM DMSO solution and 10 μL of aqueous buffer were added to 980 μL of methanol, followed by a 10-fold dilution with methanol:water (1:1). The resulting sample was analyzed by LC-MS / MS to obtain the Area (std). The kinetic solubility was calculated as follows:

[0186] Where DF is the dilution factor.

[0187] *Accurate kinetic solubility values ​​cannot be determined when the actual solubility is greater than 300 μM.

[0188] Conclusion: Compared with the control compound Urolithin A, I-8 has better solubility, overcoming the problem of poor water solubility of Urolithin A and the difficulty in developing topical preparations.

[0189] Test Example 5: Using the zebrafish model to evaluate the anti-oxidative efficacy of compounds

[0190] Experimental animals: healthy wild AB zebrafish at 48 hpf (hours post fertilization)

[0191] Evaluation of anti-oxidative efficacy: Wild-type AB zebrafish at 48 hpf were used as experimental animals, and a zebrafish oxidative aging model was established via chemical mutagenesis using 4 μM menadione as the modeling agent. Fucoxanthin was used as a positive control compound. After 22 hours of treatment with different concentrations of I-8, the fish were tested for reactive oxygen species (ROS) levels, β-galactosidase activity, and expression of three antioxidant and anti-aging-related genes (Cu / Zn-sod, Mn-sod, and AMPK), and statistical analysis was performed.

[0192] During the experiment, the modeling drug, positive compound, and I-8 were dispersed together in the culture water of zebrafish. The larvae of each group were placed in a constant temperature incubator at 28.5°C and kept away from light for 22 hours. TM Deep Red fluorescent probe and β-galactosidase staining kit were used to specifically label ROS and β-galactosidase in the fish, respectively. Image-Pro Plus software was used to quantify the staining intensity. After extracting RNA from the whole fish, fluorescent real-time quantitative PCR (RT-qPCR) was used to detect changes in the expression of three genes in the fish: Cu / Zn-sod, Mn-sod, and AMPK. GraphPad software was used to perform statistical analysis on the data. The results are presented in Figures 10 to 14.

[0193] Conclusion: As shown in Figures 10-13, compared with the blank control group, continuous treatment with menadione for 22 hours significantly increased ROS levels and β-galactosidase activity in zebrafish, with statistical significance. Menadione is an oxidant that can generate unstable semiquinones through the intracellular reductase system, which in turn generates large amounts of ROS, accelerating aging. β-galactosidase is a hydrolase found in lysosomes, and increased activity is a hallmark of cellular aging. When I-8 was applied to the zebrafish oxidative aging model at a concentration of 22.2 μg / mL, both ROS levels and β-galactosidase activity were significantly decreased compared with the control group. These results suggest that I-8, at a concentration of 22.2 μg / mL, exhibits a certain antioxidant effect.

[0194] As shown in Figure 14, when the zebrafish oxidative aging model was treated with I-8 at concentrations of 2.5, 7.5, and 22.2 μg / mL, the expression of the Cu / Zn-sod and Mn-sod genes was upregulated compared to the model group, and this was statistically significant. When the zebrafish oxidative aging model was treated with I-8 at a concentration of 2.5 μg / mL, the expression of the AMPK gene was upregulated compared to the model group, and this was statistically significant.

[0195] The above results suggest that when the action concentration of I-8 is 2.5, 7.5 and 22.2 μg / mL, it can reverse the abnormal expression of Cu / Zn-sod and Mn-sod genes induced by menadione; when the action concentration is 2.5 μg / mL, it can reverse the abnormal expression of ampk gene induced by menadione.

[0196] Inflammation is a physiological response that protects the body from various insults, such as physical injury, pathogens, exposure to toxic chemicals, and ultraviolet radiation. Early manifestations of inflammation include capillary dilation, hyperpermeability, and edema. Various inflammatory mediators, such as interleukin-1α (IL-1β), interleukin-8 (IL-8), tumor necrosis factor-α (TNF-α), and prostaglandin E2 (PGE2), play key roles in both acute and chronic inflammation. PGE2 is the most abundant prostaglandin produced in the body, and its production begins with arachidonic acid. Arachidonic acid is first converted to prostaglandin H2 (PGH2) by cyclooxygenase (COX), which is then further catalyzed by prostaglandin E synthase to produce PGE2. Ultimately, PGE2 exerts its primarily inflammatory biological functions by acting on four E-type prostaglandin (EP) receptors, EP1-4. Among them, EP2 receptors are all coupled to Gs proteins and mainly send signals through the cAMP-PKA-CREB pathway triggered by adenylate cyclase. Studies have also shown that selective small molecule antagonists can be developed by targeting EP2 receptors to alleviate downstream pathological processes mediated by EP2 receptors, thereby developing a new generation of anti-inflammatory therapies.

[0197] In addition, TRPV1 receptor is a nociceptor that can be activated by a variety of factors such as chemicals (capsaicin), noxious heat stimulation and acidification. TRPV1 is widely present in C-type sensory nerve afferent fibers and keratinocytes. After the TRPV1 receptor is activated, monovalent and divalent cations (mainly Ca 2+ ) enters the cell, triggering an action potential, which is transmitted to the higher central nervous system, producing a burning pain sensation. Therefore, after the test substance acts, blocking or inhibiting the expression of TRPV1 receptor protein helps to relieve the burning pain and achieve the purpose of soothing. Therefore, by detecting the expression level of TRPV1 gene in keratinocytes after sample treatment, this dimension indicator can be used to preliminarily determine whether the sample has a soothing effect.

[0198] In the following test examples 6-1 and 6-2, a model was established using immortalized keratinocytes (HaCaT) and UVB, and the expression levels of TNF-α, IL-8, IL-1β, and EP2 were detected after the test compound was applied to HaCaT; a model was established using immortalized keratinocytes (HaCaT) and capsaicin, and the expression level of TRPV1 was detected after the test compound was applied to HaCaT, thereby conducting a multi-dimensional evaluation of the soothing effect of the test substance.

[0199] Test Example 6-1: Detection of TNF-α, IL-8, IL-1β, and EP2 Gene Expression in HaCaT Cells after UVB Irradiation

[0200] 1) Cell Seeding: HaCaT cells were seeded at 6×105 cells / well in a 6-well plate and cultured in an incubator (37°C, 5% CO2) for 12 h. A normal control group (0 mJ / cm2 + vehicle control), a model control group (80 mJ / cm2 + vehicle control), a low-concentration group (80 mJ / cm2 + low-concentration compound), a medium-concentration group (80 mJ / cm2 + medium-concentration compound), and a high-concentration group (80 mJ / cm2 + high-concentration compound) were set up.

[0201] 2) UVB modeling: Before UVB irradiation, wash the HaCaT cells three times with D'Hanks. Add 1 mL of D'Hanks to the wells to submerge the cells. Wrap the control cells in tin foil and place them in the dark. Based on the experimental groups, perform UVB modeling (80 mJ / cm2) in each irradiation group.

[0202] 3) Administration: Each experimental group was added with DMEM medium containing different concentrations of compounds and cultured for 24 hours;

[0203] 4) After the culture is completed, the cells are sampled, total RNA is extracted from each experimental group, cDNA is synthesized, and the gene expression of β-actin and the target gene is detected by q-PCR.

[0204] 5) Using β-actin as an internal reference for gene expression, calculate the relative RNA expression of the target gene. ΔΔC(t) ΔC(t)=C(t) 目的基因 -C(t) β-actin

[0205] Test Example 6-2: Detecting TRPV1 gene expression in HaCaT cells after capsaicin stimulation

[0206] 1) Cell Seeding: HaCaT cells were seeded at 6 × 105 cells / well in a 6-well plate and cultured in an incubator (37°C, 5% CO2) for 12 h. A normal control group (0 μM capsaicin + vehicle control), a model control group (0 μM capsaicin + vehicle control), a low-concentration group (15 μM capsaicin + low-concentration compound), a medium-concentration group (15 μM capsaicin + medium-concentration compound), and a high-concentration group (15 μM capsaicin + high-concentration compound) were set up.

[0207] 2) Induction and Dosing: Discard the culture medium in the 6-well plate and proceed with dosing. Add 1 mL of culture medium containing the test substance and capsaicin stock solution to each well according to the drug grouping described above. After dosing, place the 24-well plate in an incubator (37°C, 5% CO2) and incubate for 24 h ± 2 h.

[0208] 3) After the incubation, the cells were gently rinsed once or twice with D-Hanks. Fresh culture medium was added to the normal control group, and fresh culture medium containing the corresponding concentration of compound was added to the sample group. The cells were cultured at 37° C. and 5% CO 2 for 24 h.

[0209] 4) After the culture is completed, the cells are sampled, total RNA is extracted from each experimental group, cDNA is synthesized, and the gene expression of β-actin and the target gene is detected by q-PCR.

[0210] 5) Using β-actin as an internal reference for gene expression, calculate the relative RNA expression of the target gene. ΔΔC(t) ΔC(t)=C(t) 目的基因 -C(t) β-actin

[0211] As shown in Figures 15 to 19, compound I-8 was able to downregulate the expression of TNF-α, IL-8, IL-1β, EP2, and TRPV1 in a dose-dependent manner, suggesting that compound I-8 has a soothing effect.

[0212] Test Example 7: Evaluation of the Antioxidant Efficacy of Compounds Using HaCaT Human Immortalized Keratinocytes

[0213] Oxidative reactions in organisms, such as respiratory metabolism, generate reactive free radicals. Under normal conditions, free radicals possess a stable scavenging system, maintaining a low concentration. Oxidative stress (OS) occurs when the balance between oxidative and antioxidant functions in the body is disrupted by endogenous and / or exogenous stimuli, leading to excessive free radical production. Excessive free radicals produce a range of negative effects in the body, including oxidative damage to biomolecules and further cell death and tissue damage. In vivo, except for a very small amount of ROS that is utilized by the body, virtually all ROS must be promptly cleared. Radiation damage from medium-wave erythematous ultraviolet (UVB) (280-319nm) is the most important factor in skin photoaging, primarily damaging keratinocytes (HaCaTs). This is manifested by the accumulation of photoproducts, increased ROS, and increased oxidative damage. Organisms have formed a complete set of antioxidant systems in the long-term evolution process, which keeps the production and elimination of free radicals in a dynamic balance. It is an adaptive mechanism of organisms. The main members include antioxidant enzymes, antioxidants, and proteins that separate transition metals, etc. They can all specifically limit the body's oxidative damage. Therefore, the antioxidant efficacy of the test compound can be evaluated by measuring the ROS clearance rate in keratinocytes after UVB radiation. The following test example 7 uses HaCaT human immortalized keratinocytes to evaluate the antioxidant efficacy of the compound.

[0214] Test Example 7: Detection of ROS content in HaCaT after UVB irradiation

[0215] 1) Cell seeding: HaCaT cells were seeded at 2×10 4 The cells were inoculated into 96-well plates and cultured in an incubator (37°C, 5% CO2) for 12 h. A control group (120 mJ / cm 2 + solvent control), low concentration group (120mJ / cm 2 + low concentration compound), medium concentration group (120mJ / cm 2 + medium concentration compound), high concentration group (120mJ / cm 2 + high concentration compounds);

[0216] 2) UVB modeling: Before UVB irradiation, wash the HaCaT cells three times with D'Hanks. Add 50 μL of D'Hanks to the wells to submerge the cells. Wrap the control cells in tin foil and place them in the dark. UVB modeling was performed on each irradiation group.

[0217] 3) Administration: Each experimental group was added with DMEM medium containing different concentrations of compounds and cultured for 24 hours;

[0218] 4) ROS fluorescent probe production: Dilute DCFH-DA with PBS at a 1:1000 dilution ratio to a final concentration of 10 μmol / L. Discard the culture medium from all wells except the wells containing bare cells. Wash three times with PBS, then add 200 μL of DCFH-DA working solution to each well. Incubate in a CO2 incubator for 30 min. After incubation, rinse each well three times with PBS.

[0219] 5) Fluorescence analysis: Place the 96-well plate to be tested on the fluorescence microplate reader, set the incident light wavelength to 525 nm and the excitation light wavelength to 488 nm, and read the results.

[0220] Experimental results: As shown in Figure 20, compound I-8 can scavenge ROS in a dose-dependent manner, indicating that compound I-8 has antioxidant effects.

[0221] With aging, human skin gradually experiences atrophy (thinning), fragility, poor pigmentation, and delayed wound healing. Skin fragility is attributed in part to changes in hemidesmosomes and downregulation of the expression of various collagens (Collagen I, III, IV, VII) or laminins (such as LN-5) at the dermal-epidermal junction. LN-5 has been shown to be a component of anchoring fibers in the basement membranes of the skin, cornea, conjunctiva, and other tissues. LN-5 participates in cell-to-cell interactions through the mediation of integrins and proteoglycans, playing a crucial role in cell adhesion, growth, migration, and differentiation. The appearance of skin wrinkles is closely related to the normal synthesis and expression of collagen and laminin. Therefore, increasing the aforementioned collagen content and LN-5 can achieve a certain degree of wrinkle resistance and play an important role in the aging process of the skin. Therefore, the firming and anti-wrinkle efficacy of the test compounds was evaluated by measuring Collagen I protein levels in epidermal cells (fibroblasts) using ELISA, and by measuring transcript levels in epidermal cells (keratinocytes) III, IV, VII, and LN-5 using real-time PCR. The following Test Examples 8-1 and 8-2 evaluated the anti-wrinkle efficacy of the compounds using HaCaT immortalized human keratinocytes and HSF human skin fibroblasts.

[0222] Test Example 8-1: Detection of Type I Collagen Expression in HSF Human Skin Fibroblasts

[0223] 1) Cell seeding: 2×10 cells were seeded into 96-well plates. 4 cells / well (37°C, 5% CO2) and cultured for 24 hours.

[0224] 2) Dosing: Discard the culture medium in the 96-well plate and proceed with dosing. Add the culture medium containing the compound to the sample group and the cell culture medium without the compound to the control group, 200 μL per well. After dosing, place the 96-well plate in an incubator (37°C, 5% CO2) for 24 h ± 2 h.

[0225] 3) Type I collagen detection: After the incubation, the cell supernatant was collected and the type I collagen content was determined using a human type I collagen enzyme-linked immunosorbent assay kit.

[0226] Test Example 8-2: Detection of Collagen III, IV, VII, and LN-5 Gene Expression Levels in HaCaT Human Immortalized Keratinocytes

[0227] 1) Cell seeding: Epidermal cells were seeded at 1×10 6 Cells / well were seeded in 6-well plates and cultured in an incubator (37°C, 5% CO2) for 12 h. A control group (0% compound), a low-concentration group (low-concentration compound), a medium-concentration group (medium-concentration compound), and a high-concentration group (high-concentration compound) were set up.

[0228] 2) Administration: According to the experimental groups, add DMEM medium containing different concentrations of compounds and continue culturing for 24 hours;

[0229] 3) Cell collection: After the culture is completed, cell samples are collected for subsequent detection of Collagen III, IV, VII and LN-5 expression.

[0230] As shown in Figures 21 to 25 , compound I-8 can increase the secretion of type I collagen and upregulate the expression of COL3A1, COL4A1, COL7A1, and LAMA5 genes in a dose-dependent manner, suggesting that compound I-8 has anti-wrinkle efficacy.

[0231] Test Example 9: Detection and calculation of the ROS clearance rate of each compound on HaCaT cells after UVB irradiation

[0232] Using the experimental method in Test Example 7, the ROS scavenging rate of each compound on HaCaT cells after UVB irradiation was calculated according to the following formula. The results are listed in Table 1.

[0233] Where S: fluorescence intensity

[0234] Table 1.

[0235] Those skilled in the art will appreciate that the above-mentioned embodiments are specific examples for implementing the present invention, and that in actual applications, various changes may be made thereto in form and detail without departing from the spirit and scope of the present invention.

Claims

1. A compound having a structure as shown in the following general formula I, a salt, a stereoisomer or a solvate thereof, in, X is oxygen or nothing; R 1 and R 2 are independently hydrogen, hydroxyl, C 1-6 Alkyl, C 1-6 Alkoxy, amino, -NC(O)R 1-1 、-NR a R b 、-CH2C(O)OR 1-2 , phenyl, -OC(O)R 1-3 、-OCH2OC(O)R 1-4 , halogen, cyano, nitro, -C(O)R 1-5 or -COOR 1-6 ; R 1-1 , R 1-2 , R 1-3 , R 1-4 , R 1-5 and R 1-6 are independently hydrogen, C 1-6 Alkyl, halogen substituted C 1-6 Alkyl or -NR a R b ; R a and R b are independently hydrogen, C 1-6 Alkyl or halogen substituted C 1-6 alkyl.

2. The compound, salt, stereoisomer or solvate thereof according to claim 1, characterized in that X is oxygen.

3. The compound, salt, stereoisomer or solvate thereof according to claim 1, characterized in that The C 1-6 Alkyl is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl; and / or, the C 1-6 Alkoxy is methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy or tert-butoxy; and / or, the halogen is fluorine, chlorine, bromine or iodine; and / or, the halogen-substituted C 1-6 The alkyl group is halogen-substituted methyl, halogen-substituted ethyl, halogen-substituted n-propyl, halogen-substituted isopropyl, halogen-substituted n-butyl, halogen-substituted isobutyl or halogen-substituted tert-butyl.

4. The compound, salt, stereoisomer or solvate thereof according to claim 1, characterized in that R 1 and R 2 are independently hydrogen, hydroxyl, C 1-6 Alkyl, C 1-6 Alkoxy, amino, -NC(O)R 1-1 、-NR a R b 、-CH2C(O)OR 1-2 , phenyl, -OC(O)R 1-3 or -OCH2OC(O)R 1-4 .

5. The compound, salt, stereoisomer or solvate thereof according to claim 1, characterized in that: R 1 and R 2 are independently hydrogen, hydroxyl, C 1-6 Alkoxy, Among them, R 1-3 and R 1-4 are independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl or 6. The compound, salt, stereoisomer or solvate thereof according to claim 1, characterized in that: R 1 and R 2 are independently hydroxy, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, 7. The compound, salt, stereoisomer or solvate thereof according to any one of claims 1 to 6, characterized in that The compound is selected from any one of the following:

8. A composition, characterized in that The composition comprises the compound according to any one of claims 1 to 7, its salt, stereoisomer or solvate; and cosmetically acceptable excipients.

9. Use of the compound according to any one of claims 1 to 7, its salt, stereoisomer or solvate, or the composition according to claim 8 in the preparation of cosmetics.

10. A method for relieving skin inflammation and / or reducing active oxygen in skin cells, characterized in that: The method comprises the steps of: The compound, salt, stereoisomer or solvate thereof according to any one of claims 1 to 7 is administered to the subject; or the composition according to claim 8 is administered to the subject.

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