Mild anti-aging composition as well as preparation method, cosmetics and application thereof

Abstract

The invention provides a mild anti-aging composition as well as a preparation method, cosmetics and application thereof, and belongs to the technical field of cosmetics. The mild anti-aging composition disclosed by the invention is prepared from hydroxyl pinacolone 9-cis retinoate and Gravi-A, wherein the Gravi-A is prepared from hydroxyl pinacolone retinoate and retinol propionate according to a mass ratio of 5 to 9. The composition disclosed by the invention can have a synergistic anti-aging effect in a specific proportion range; particularly, when the mass ratio of the hydroxyl pinacolone 9-cis-retinoate to the Gravi-A is 8: 2, under the condition that no anti-irritation component is added, the expression of molecular targets of inflammatory reaction can be effectively reduced, so that the irritation to the skin is reduced. The composition not only realizes an excellent anti-aging effect, but also considers the mild demand of skin, reduces the dosage of hydroxypinacolone 9-cis-retinoate, reduces the cost, and has a wide prospect when being applied to preparation of cosmetics.

Description

Technical Field

[0001] The present invention relates to the technical field of cosmetics, and in particular to a mild anti-aging composition, a preparation method thereof, cosmetics and applications thereof. Background Art

[0002] Skin aging, as an early indicator of systemic aging, has long been a focus of researchers in the field of aging. Beyond the inevitable natural aging process, how to effectively delay skin aging has long been a concern for dermatologists and skincare product developers. Early manifestations of skin aging include sagging, wrinkles, dry skin, and a dull complexion. With age, skin elasticity and collagen decrease, leading to fine lines on the face, especially in areas of high expression such as around the eyes, forehead, and corners of the mouth. Skin aging can also lead to uneven pigmentation, resulting in spots or freckles, making the complexion appear darker.

[0003] As early as the 1980s, studies have confirmed that vitamin A can improve skin aging. In-depth research on anti-aging products has revealed the positive effects of topical vitamin A in reducing the damage caused by aging. However, the potential irritation caused by vitamin A, such as erythema, stinging, or burning, has limited its wider application. Therefore, in the study of anti-aging products, researchers have developed a variety of vitamin A derivatives, such as retinol, retinal, and retinoic acid. Retinoic acid is considered one of the most effective substances in slowing the aging process.

[0004] Hydroxypinacolone Retinoate (HPR) is a retinoic acid derivative formed by the dehydration condensation of retinoic acid and pinacol. It retains the functional groups of retinoic acid and can act directly on retinoic acid receptors without converting to retinoic acid. This allows it to demonstrate excellent anti-aging, sebum suppression, and whitening and spot-removing properties. As a new generation of vitamin A derivatives, HPR offers advantages over previous derivatives in terms of stability, safety, and efficacy. Although HPR is significantly less irritating than retinol, its inclusion level in cosmetics is subject to certain restrictions. Excessive addition may increase product irritation and compromise product stability.

[0005] Hydroxypinacolone 9-cis retinoate has an isomerized chemical structure relative to that of HPR. CN116947723 A discloses hydroxypinacolone 9-cis retinoate, its preparation method, and applications. This patent describes hydroxypinacolone 9-cis retinoate, which exhibits significantly reduced irritation and significantly improved skin anti-aging efficacy compared to HPR. Adding hydroxypinacolone 9-cis retinoate to cosmetics can reduce the skin irritation of retinoids while enhancing their anti-aging efficacy. However, due to its high price, adding hydroxypinacolone 9-cis retinoate alone to cosmetics can significantly increase costs, resulting in a lack of economic benefit.

[0006] Therefore, it is very meaningful to develop a product that can reduce the usage of hydroxypinacolone 9-cis retinoic acid ester and has a synergistic anti-aging effect while being mild and non-irritating. Summary of the Invention

[0007] In view of the deficiencies in the prior art, the present invention discloses a mild anti-aging composition, a preparation method thereof, a cosmetic and applications thereof.

[0008] In one aspect, the present invention provides a mild anti-aging composition comprising hydroxypinacolone 9-cis retinoate (abbreviated as 111) and Gravi-A, wherein Gravi-A is composed of hydroxypinacolone retinoate (abbreviated as HRP) and retinol propionate (abbreviated as RP).

[0009] Based on extensive experimental research, the research and development team of this invention combined 111 and Gravi-A. Experimental testing of the mRNA expression of their anti-aging targets (type I collagen, type III collagen, type IV collagen, and elastin) and retinol metabolism targets (RAR, RXR, and CRBPI) revealed that within a specific ratio, 111 and Gravi-A in the combination synergistically promoted gene expression, achieving an anti-aging effect. Simultaneously, by testing the inflammatory target IL-1α, the research and development team unexpectedly discovered that the present composition, without the addition of anti-irritant components, can effectively reduce the expression of molecular targets of the inflammatory response, thereby reducing skin irritation. Therefore, the mild anti-aging composition of the present invention not only achieves excellent anti-aging effects, but also takes into account the skin's gentleness requirements, reduces the amount of hydroxypinacolone 9-cis retinoate used, and reduces costs. Its application in the preparation of cosmetics has broad prospects.

[0010] Furthermore, the mass ratio of hydroxypinacolone retinoic acid ester to retinol propionate is 5:9.

[0011] Patent CN 116947723 A, filed by the research and development team of this invention, discloses a synergistic effect achieved by combining hydroxypinacolone retinoate (HPR) and retinyl propionate (RP) in a mass ratio of (5-40):9. This synergistic effect is particularly pronounced when the mass ratio of hydroxypinacolone retinoate to retinyl propionate is 5:9, significantly increasing the expression of key aging-related proteins Col-IV, CRBPI, RARB, and Elastin. The expression of type IV collagen was increased by more than 4-fold, CRBP-1 by more than 30-fold, RARB by more than 5-fold, and Elastin by more than 300-fold, without any cytotoxicity.

[0012] Furthermore, the mass ratio of hydroxypinacolone 9-cis retinoic acid ester to Gravi-A is (1-8): (2-9).

[0013] Within the above-mentioned mass ratio range, 111 and Gravi-A of the present invention have the ability to synergistically promote the expression of anti-aging targets (type I collagen, type III collagen, type IV collagen and elastin) and retinol metabolism targets (RAR, RXR and CRBPI) at the mRNA level, thereby achieving the anti-aging effect of the skin.

[0014] Furthermore, the mass ratio of hydroxypinacolone 9-cis retinoic acid ester to Gravi-A is (1-5): (5-9).

[0015] The present invention's 111 and Gravi-A, within the aforementioned mass ratio range, can synergistically increase the expression of a series of important anti-aging-related proteins while simultaneously reducing the use of expensive hydroxypinacolone 9-cis retinoic acid ester by at least 50%. Exemplarily, the mass ratio of 111 to Gravi-A is 1:9; 2:8; or 5:5. The present embodiments provide the relevant components of the mild anti-aging composition.

[0016] Furthermore, the mass ratio of hydroxypinacolone 9-cis retinoic acid ester to Gravi-A is 8:2.

[0017] When the mild anti-aging composition has the above mass ratios, it not only promotes the expression of collagen, an anti-aging target, but also promotes the expression of retinol metabolism targets, and effectively reduces the expression of molecular targets of inflammatory response, thus achieving a good balance between excellent anti-aging efficacy and excellent mildness. The examples of the present invention provide the relevant content of the components of the mild anti-aging composition.

[0018] In another aspect, the present invention also provides a method for preparing a mild anti-aging composition, wherein the components of the composition are mixed in proportion, a solvent is added, and the mixture is heated and stirred to obtain the mild anti-aging composition.

[0019] The heating and stirring temperature of the present invention does not exceed 80°C, and the solvent is any one of an organic solvent or a grease solvent.

[0020] The organic solvent is one or more of dimethyl sulfoxide, isosorbide dimethyl ether, 2,3-butanediol, bis-diethoxydiglycol cyclohexane 1,4-dicarboxylate, ethoxydiglycol, and triethyl citrate.

[0021] The oil solvent is any one or a combination of the following raw materials: soybean oil, olive oil, almond oil, castor oil, peanut oil, cottonseed oil, tea seed oil, jojoba oil, avocado oil, coconut oil, palm oil, cocoa butter, rice bran oil, evening primrose oil, wheat germ oil, corn germ oil, palm wax, wood wax, mink oil, turtle oil, snake oil, egg yolk oil, beef tallow, horse tallow, lard, Deer fat, lecithin, lanolin, beeswax, spermaceti, shellac wax, paraffin, petrolatum, microcrystalline wax, ozokerite, squalane, lanolin wax, lanolin alcohol, lanolin alcohol esters, acetylated lanolin, acetylated lanolin alcohol, lanolin acid, lanolin acid esters, polyoxyethylene lanolin, polyoxyethylene lanolin alcohol ether, polyoxypropylene lanolin alcohol ether, hydrogenated lanolin, alkoxylated hydrogenated lanolin, silicone oil (such as dimethyl silicone oil, octamethyl silicone oil, methylphenyl polysiloxane), polysiloxane-polyoxyalkyl block copolymer, methyl hydrogen silicone oil, Cn fatty acids and esters thereof, Cn fatty alcohols and esters thereof; wherein n is ≥10, and is preferably 10-24, more preferably 12-20, such as 14, 16, 17, and 18. The Cn fatty acids include lauric acid, myristic acid, palmitic acid, stearic acid, oleic acid, linoleic acid, erucic acid, and capric acid. The Cn fatty alcohols include lauryl alcohol, cetyl alcohol, stearyl alcohol, isostearyl alcohol, oleyl alcohol, and coconut oil. The Cn fatty acid esters and Cn fatty alcohol esters include isopropyl myristate, myristyl myristate, myristyl lactate, cetyl lactate, isopropyl palmitate, isooctyl palmitate, butyl stearate, isooctyl stearate, monoglyceride stearate, polyethylene glycol stearate, decyl oleate, glyceryl caprylate, glyceryl caprate, and triolein.

[0022] In another aspect, the present invention also provides a cosmetic comprising a mild anti-aging composition.

[0023] Furthermore, the cosmetics include any one of water, lotion, cream, mask, gel, essence and essence milk.

[0024] Furthermore, the cosmetics also include moisturizers, emulsifiers, thickeners, preservatives, pH regulators, other active ingredients, etc.

[0025] Exemplarily, the moisturizing agent includes one or more of propylene glycol, 1,3-propylene glycol, methylpropylene glycol, glycerin, diglycerin, butylene glycol, 2,3-butylene glycol, 1,2-pentanediol, 1,2-hexanediol, dipropylene glycol, dimethyl isosorbide, 2,3-butylene glycol, bis-diethoxydiglycol cyclohexane 1,4-dicarboxylate, ethoxydiglycol, and triethyl citrate. The mass percentage of the moisturizing agent in the cosmetic is 0.1-10%.

[0026] Exemplarily, the emulsifier includes one or more of glyceryl stearate, PEG-100 stearate, cetearyl alcohol, ceteareth-20, PEG-40 stearate, cetearyl glucoside, C14-22 alcohol, C12-20 alkyl glucoside, polyglyceryl-3 methylglucose distearate, polyglyceryl-10 dipalmitate, polyglyceryl-3 stearate, C20-22 alcohol phosphate, C20-22 alcohol, potassium cetyl phosphate, sodium stearoyl glutamate, sodium cetearyl sulfate, sodium stearoyl lactylate, cetearyl olivate, sorbitan olivate, sucrose polystearate, cetyl palmitate, olive oil PEG-6 polyglyceryl-6 esters, lecithin, and hydrogenated lecithin. The mass percentage of the emulsifier in the cosmetic is 0.1 to 5%.

[0027] Exemplary thickeners include one or more of hydroxyethyl cellulose, hydroxypropyl methylcellulose, xanthan gum, sodium hydroxypropyl starch phosphate, acrylates / C10-30 alkyl acrylate crosspolymer, acrylates copolymer, sodium acrylates copolymer, carbomer, ammonium acryloyldimethyltaurate / VP copolymer, and sodium polyacryloyldimethyltaurate. The mass percentage of thickeners in cosmetics ranges from 0.1% to 1%.

[0028] For example, the preservative includes one or more of phenoxyethanol, ethylhexylglycerin, caprylyl glycol, and p-hydroxyacetophenone. The mass percentage of the preservative in the cosmetic is in the range of 0.3-0.6%.

[0029] For example, the chelating agent includes one or more of disodium edetate, sodium phytate, tetrasodium glutamate diacetate, disodium dihydrogen pyrophosphate, sodium citrate, and trisodium edetate. The mass percentage of the chelating agent in the cosmetic is in the range of 0.05-0.1%.

[0030] Exemplarily, the pH regulator includes one or more of sodium hydroxide, potassium hydroxide, triethanolamine, tromethamine, and arginine. The pH regulator is present in a cosmetic in an amount ranging from 0.1% to 1% by weight.

[0031] For example, the other active ingredients include one or more of tocopherol (vitamin E), tocopherol acetate, ectoine, niacinamide, etc. The mass percentage of the other active ingredients in the cosmetic is in the range of 0.1-1%.

[0032] Furthermore, the mass percentage of the anti-aging composition in the preparation of cosmetics is 0.01-0.3%.

[0033] On the other hand, the present invention also provides an application of a mild anti-aging composition in the preparation of cosmetics. In actual application, the mass ratio of 111 and Gravi-A in the composition can be selected according to different application scenarios.

[0034] Compared with the prior art, the present invention has the following advantages: the mild anti-aging composition of the present invention exhibits a synergistic anti-aging effect within a specific ratio range. Furthermore, without the addition of anti-irritant components, the composition can effectively reduce the expression of molecular targets of inflammatory responses, thereby reducing skin irritation. Furthermore, the composition reduces the amount of hydroxypinacolone 9-cis retinoate used, lowering production costs. Its application in the preparation of cosmetics has broad prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0035] The accompanying drawings, which constitute part of this application, are intended to provide a further understanding of the present invention. The exemplary embodiments of the present invention and their descriptions are intended to explain the present invention and do not constitute an undue limitation of the present invention. In the accompanying drawings: Figure 1 This is a graph showing the effects of different proportions of mild anti-aging compositions on Col 1 mRNA expression in the anti-aging target test of Test Example 1; Figure 2 This is a graph showing the effects of different proportions of mild anti-aging compositions on Col Ⅲ mRNA expression in the anti-aging target test of Test Example 1; Figure 3 This is a graph showing the effects of different proportions of mild anti-aging compositions on Col IV mRNA expression in the anti-aging target test of Test Example 1; Figure 4 This is a graph showing the effects of different proportions of mild anti-aging compositions on Elastin mRNA expression in the anti-aging target test of Test Example 1; Figure 5 This is a graph showing the effects of different proportions of mild anti-aging compositions on RARA mRNA expression in the A-alcohol metabolism target test in Test Example 2; Figure 6 This is a graph showing the effects of different proportions of mild anti-aging compositions on RARB mRNA expression in the A-alcohol metabolism target test in Test Example 2; Figure 7 This is a graph showing the effects of different proportions of mild anti-aging compositions on RARG mRNA expression in the A-alcohol metabolism target test in Test Example 2; Figure 8 This is a graph showing the effects of different proportions of mild anti-aging compositions on RXRA mRNA expression in the A-alcohol metabolism target test in Test Example 2; Figure 9 This is a graph showing the effects of different proportions of mild anti-aging compositions on RXRB mRNA expression in the A-alcohol metabolism target test in Test Example 2; Figure 10 This is a graph showing the effects of different proportions of mild anti-aging compositions on RXRG mRNA expression in the A-alcohol metabolism target test in Test Example 2; Figure 11 This is a graph showing the effects of different proportions of mild anti-aging compositions on CRBP-1 mRNA expression in the A-alcohol metabolism target test in Test Example 2; Figure 12 This is a graph showing the effects of different proportions of mild anti-aging compositions on IL-1α expression in the inflammatory target test of Test Example 3; Figure 13 This is a graph showing the effect of hydroxypinacolone 9-cis retinoate on the survival rate of human fibroblasts in the cytotoxicity test of Test Example 4; Figure 14 This is a graph showing the effect of hydroxypinacolone retinoate on the survival rate of human fibroblasts in the cytotoxicity test of Test Example 4; Figure 15 This is a graph showing the effect of retinol propionate on the survival rate of human fibroblasts in the cytotoxicity test of Test Example 4; Figure 16 This is the evaluation result of the serum lotion in Example 3 on the length of forehead wrinkles in the human anti-aging evaluation in Test Example 5; Figure 17 This is the evaluation result of the essence lotion of Example 3 on the length of glabellar lines in the human anti-aging evaluation of Test Example 5; Figure 18 This is the evaluation result of the crow's feet length of the essence lotion in Example 3 in the human anti-aging evaluation of Test Example 5; Figure 19 This is the result of the evaluation of the length of nasolabial folds by the essence lotion of Example 3 in the human anti-aging evaluation of Test Example 5; Figure 20 This is the evaluation result of the neck wrinkle length by the essence lotion of Example 3 in the human anti-aging evaluation of Test Example 5; Figure 21 This is the evaluation result of the length of corner wrinkles of the mouth using the essence lotion of Example 3 in the human anti-aging evaluation of Test Example 5; Figure 22 This is the result of the evaluation of the skin glossiness of the essence lotion in Example 3 in the human anti-aging evaluation of Test Example 5; Figure 23 This is the result of the evaluation of the number of pores by the essence lotion in Example 3 in the human anti-aging evaluation of Test Example 5; Figure 24 This is the evaluation result of the essence lotion of Example 3 on cheek skin wrinkles in the human anti-aging evaluation of Test Example 5; Figure 25 This is a graph showing the evaluation results of the skin stratum corneum moisture content of the essence lotion in Example 3 in the human anti-aging evaluation of Test Example 5; Figure 26 This is a graph showing the evaluation results of the skin firmness parameter F4 of the essence lotion in Example 3 in the human anti-aging evaluation of Test Example 5; Figure 27 This is a graph showing the evaluation results of the skin elasticity parameter R2 of the essence lotion in Example 3 in the human anti-aging evaluation of Test Example 5; Figure 28 This is a graph showing the evaluation results of the skin elasticity parameter R5 of the essence lotion in Example 3 in the human anti-aging evaluation of Test Example 5. DETAILED DESCRIPTION

[0036] In order to clearly and completely describe the technical solutions in the embodiments of the present invention below, it is obvious that the embodiments described are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention.

[0037] To facilitate understanding of the present invention, the present invention will be described in more detail below, with preferred embodiments of the present invention provided. However, it should be understood that these embodiments are provided for further explanation only and are not intended to limit the present invention in any manner, i.e., they do not limit the scope of protection of the present invention.

[0038] The present invention will be further described below with reference to the embodiments.

[0039] The various components of the mild anti-aging composition of the present invention are described in detail.

[0040] Hydroxypinacolone 9-cis retinoate Hydroxypinacolone 9-cis retinoate. CN 116947723 A discloses the preparation method and application of this substance. The patent tests the expression levels of aging-related genes and RAR / RXR receptors, proving that hydroxypinacolone 9-cis retinoate has reduced irritation and improved skin anti-aging efficacy compared to HPR.

[0041] The mild anti-aging composition of the present invention comprises hydroxypinacolone 9-cis retinoic acid ester.

[0042] The structure of hydroxypinacolone 9-cis retinoic acid ester is as follows:

[0043] The hydroxypinacolone 9-cis retinoic acid ester used in the present invention is prepared by the preparation method of CN 116947723 A, specifically, the following method: (1) Dissolve 9-cis retinoic acid in a dimethylformamide solution, and add cesium carbonate and 1-chloropinacolone in sequence; (2) stirring the reaction mixture obtained in step (1) at room temperature, adding deionized water to quench the mixture, and then extracting the product with dichloromethane; (3) Combine the organic extracts, dry them, and then concentrate them under reduced pressure; (4) Purify by silica gel column chromatography using a hexane / ethyl acetate mixture with a volume ratio of 5:1.

[0044] Hydroxypinacolone retinoate Hydroxypinacolone retinoate is a novel retinoic acid derivative formed by the dehydration condensation of retinoic acid and pinacol. It retains the functional groups of retinoic acid and can act directly on retinoic acid receptors without converting to retinoic acid. This ester exhibits excellent anti-aging, sebum suppression, and whitening and spot-removing properties.

[0045] The mild anti-aging composition of the present invention comprises hydroxypinacolone retinoate.

[0046] The structure of hydroxypinacolone retinoate is as follows:

[0047] Hydroxypinacolone retinoic acid ester can be used without particular limitation as long as it is, for example, a hydroxypinacolone retinoic acid ester commonly used in cosmetics.

[0048] As hydroxypinacolone retinoic acid ester, a commercially available product can be used.

[0049] Retinyl Propionate The mild anti-aging composition of the present invention comprises retinyl propionate.

[0050] The structure of retinyl propionate is as follows:

[0051] Retinol propionate can be used without particular limitation as long as it is a retinol propionate commonly used in cosmetics.

[0052] As retinyl propionate, a commercially available product can be used.

[0053] The anti-aging targets, A-alcohol metabolism targets, and inflammatory targets of the test experiments of the embodiments of the present invention include the following specific contents: The instruments, kits, cells, and reagents used in the test experiments of the embodiments of the present invention can all be purchased through public commercial channels and are commonly used in the field. The operating procedures of the instruments and kits are carried out according to the instructions of their instructions.

[0054] Anti-aging target type I collagen Type I collagen (Col I) is the most abundant type of collagen in the skin and provides structure and strength to the skin. It forms the main fiber network in the dermis layer of the skin, maintaining the skin's stability and elasticity.

[0055] Anti-aging target type III collagen Type III collagen (Col III) exists in the skin as fine fibers, interwoven with type I collagen, providing suppleness and elasticity to the skin. It helps maintain skin's flexibility and reduce the formation of wrinkles.

[0056] Anti-aging target type IV collagen Type IV collagen (Col IV) is the main type of collagen that makes up the basement membrane zone (BMZ) of the skin. It forms the BM's network structure, a crucial connecting layer between the epidermis and dermis, and is crucial for maintaining skin integrity and function.

[0057] Anti-aging target elastin Elastin, along with collagen fibers, forms the skin's extracellular matrix, providing structural support. Elastin forms a complex network within the dermis, interacting with collagen fibers to maintain the skin's shape and stability. Maintaining elastin is crucial for anti-aging and maintaining youthful skin.

[0058] A-alcohol metabolic targets RAR and RXR Retinol is first converted into retinoic acid (RA) in the body, a process that is crucial for retinol to exert its biological activity. Retinoic acid then binds to retinoic acid receptors (RAR and RXR) within the cell, forming a dimeric complex. This complex then binds to the retinoic acid response element (RARE or RXRE), regulating the activation and expression of mRNA in the cell nucleus. Binding to RAR and RXR promotes the maturation and differentiation of skin cells, which is crucial for maintaining normal skin renewal and repair. Furthermore, activation of RAR and RXR increases collagen synthesis and reduces collagen degradation, thereby reducing wrinkles and improving skin elasticity.

[0059] A-alcohol metabolic target CRBPI CRBPI indirectly regulates the rate and direction of retinol metabolism within cells through its ability to bind to retinol. By binding to retinol, CRBPI can affect the conversion of retinol to retinoic acid (RA), thereby regulating the concentration and activity of RA within cells.

[0060] Inflammatory target IL-1α IL-1α is a potent cytokine with potent inflammatory and immune-amplifying effects, primarily expressed by various cell types in response to external stimuli. While alcohols often promote IL-1α expression, the composition of the present invention can reduce IL-1α expression.

[0061] Example 1 Preparation of composition Mild anti-aging composition 1: (Gravi-A / 111=9:1) 1 g of hydroxypinacolone 9-cis retinoate, 3.214 g of hydroxypinacolone retinoate, and 5.786 g of retinol propionate were mixed in proportion, 190 g of dimethyl sulfoxide was added, and the mixture was heated to no more than 80° C. and stirred evenly, and then cooled to obtain a mild anti-aging composition 1.

[0062] Mild anti-aging composition 2: (Gravi-A / 111=8:2) 2 g of hydroxypinacolone 9-cis retinoate, 2.857 g of hydroxypinacolone retinoate, and 5.143 g of retinol propionate were mixed in proportion, 190 g of dimethyl sulfoxide was added, and the mixture was heated to no more than 80° C. and stirred evenly, and then cooled to obtain a mild anti-aging composition 2.

[0063] Mild anti-aging composition 3: (Gravi-A / 111=5:5) 5 g of hydroxypinacolone 9-cis retinoate, 1.786 g of hydroxypinacolone retinoate, and 3.214 g of retinol propionate were mixed in proportion, 190 g of dimethyl sulfoxide was added, and the mixture was heated to no more than 80° C. with stirring, and then cooled to obtain a mild anti-aging composition 3.

[0064] Mild anti-aging composition 4: (Gravi-A / 111=2:8) 8 g of hydroxypinacolone 9-cis retinoate, 0.714 g of hydroxypinacolone retinoate, and 1.286 g of retinol propionate were mixed in proportion, 190 g of dimethyl sulfoxide was added, and the mixture was heated to no more than 80° C. and stirred evenly, and then cooled to obtain a mild anti-aging composition 4.

[0065] Mild anti-aging composition 5: (Gravi-A / 111=4:6) 6 g of hydroxypinacolone 9-cis retinoate, 1.429 g of hydroxypinacolone retinoate, and 2.571 g of retinol propionate were mixed in proportion, 190 g of dimethyl sulfoxide was added, and the mixture was heated to no more than 80° C. and stirred evenly, and then cooled to obtain a mild anti-aging composition 5.

[0066] Mild anti-aging composition 6: (Gravi-A / 111=3:7) 7 g of hydroxypinacolone 9-cis retinoate, 1.071 g of hydroxypinacolone retinoate, and 1.929 g of retinol propionate were mixed in proportion, 190 g of dimethyl sulfoxide was added, and the mixture was heated to no more than 80° C. and stirred evenly, and then cooled to obtain a mild anti-aging composition 6.

[0067] Example 2 Preparation of Comparative Example Comparative Example 1: (Gravi-A / 111=1:9) 9 g of hydroxypinacolone 9-cis retinoate, 0.357 g of hydroxypinacolone retinoate and 0.643 g of retinol propionate were mixed in proportion, 190 g of dimethyl sulfoxide was added, the mixture was heated to no more than 80° C. and stirred evenly, and the mixture was cooled to obtain Comparative Example 1.

[0068] Comparative Example 2: (Gravi-A) 3.571 g of hydroxypinacolone 9-cis retinoic acid ester and 6.429 g of retinol propionate were mixed in proportion, 190 g of dimethyl sulfoxide was added, and the mixture was heated to no more than 80° C. and stirred evenly, and then cooled to obtain Comparative Example 2.

[0069] Comparative Example 3: (111) 10 g of hydroxypinacolone 9-cis retinoic acid ester was added to 190 g of dimethyl sulfoxide, heated to no more than 80 ° C. and stirred evenly, and then cooled to obtain Comparative Example 3.

[0070] Test Example 1 Anti-aging target test The anti-aging target test of the present invention includes type I collagen (Col I), type III collagen (Col III), type IV collagen (Col IV), and elastin.

[0071] The experimental process of anti-aging target testing is as follows: human primary fibroblasts in the exponential growth phase are prepared into single cell suspensions, and 1.0-1.2×10 5Cells were seeded in 6-well plates at a density of 100 μg / mL in a final culture volume of 2 mL and cultured at 37°C in a medium containing 10% fetal bovine serum until the cells adhered. Primary fibroblasts were seeded in low-adhesion culture plates and cultured in a medium containing 10% fetal bovine serum at 37°C for 3-5 days to form spheroids. After cell attachment or spheroid formation, the supernatant was aspirated and discarded. Freshly prepared 0.075 μg / mL of the composition of Examples 1 and 2 (serum-free) was added to each well and incubated for 48 hours. Col I mRNA, Col III mRNA, Col IV mRNA, and Elastin mRNA were extracted from each well according to the standard RNA extraction protocol. The RNA was reverse-transcribed into cDNA for qPCR detection of key gene expression, and the remaining RNA was aliquoted and stored.

[0072] The above steps of RNA extraction and qPCR detection of Col I mRNA, Col III mRNA, Col IV mRNA, and Elastin mRNA expression were completed using kits commonly used by those skilled in the art, and each operation step was performed with reference to the kit instructions.

[0073] Col I, Col III, Col IV, and Elastin mRNA expression assay: Total RNA was extracted from cell samples using a Trizol RNA extraction kit and reverse transcribed into cDNA using reverse transcriptase. Specific primers were used to amplify the cDNA of the target gene, and the amplified product was monitored in real time by adding a fluorescent dye. The intensity of the fluorescent signal is proportional to the mRNA level of the target gene. Relative expression was calculated using normalization with the internal reference gene GAPDH.

[0074] Data analysis: Statistical analysis software was used for statistical analysis of the data. The experimental results were expressed as mean ± SD. The t-test was used for comparison between groups. "*" indicates p < 0.05, indicating that there was a significant difference compared with the Control, Gravi-A and 111 groups.

[0075] The test results of the expression of anti-aging targets Col I mRNA, Col III mRNA, Col IV mRNA, and Elastin mRNA are shown in Table 1.

[0076]

[0077] From Table 1 and Figure 1-4The data show that in the Col I and Col III tests, when the mass ratio of Gravi-A / 111 is 2:8 (mild anti-aging composition 4), it has significant differences compared with only Gravi-A (Comparative Example 2) or 111 (Comparative Example 3), and therefore has the effect of synergistically promoting the expression of Col I and Col III; in the Col IV test, when the mass ratio of Gravi-A / 111 is 9:1 (mild anti-aging composition 1), 8:2 (mild anti-aging composition 2), 5:5 (mild anti-aging composition 3), and 2:8 (mild anti-aging composition 4), it has significant differences compared with only Gravi-A or 111, and therefore has the effect of synergistically promoting the expression of Col IV; in the elastin test, when the mass ratio of Gravi-A / 111 is 8:2 (mild anti-aging composition 2), 5:5 (mild anti-aging composition 3), and 2:8 (mild anti-aging composition 4), it has the effect of synergistically promoting the expression of elastin compared with only Gravi-A or 111.

[0078] In summary, both Gravi-A and 111 have the effect of promoting the expression of anti-aging targets, but only when the two substances are combined in a specific ratio can they have a synergistic effect. In some ratios, the compound even performs worse than Gravi-A or 111. For example, when the mass ratio of Gravi-A / 111 is 1:9 (Comparative Example 1), the Col IV test result of Comparative Example 1 is lower than that of Comparative Example 2 (Gravi-A) and Comparative Example 3 (111).

[0079] Test Example 2: A-alcohol metabolism target test The A-alcohol metabolism target test of the present invention includes RARA mRNA, RARB mRNA, RARG mRNA, RXRA mRNA, RXRB mRNA, RXRG mRNA, and CRBP-I mRNA.

[0080] The experimental process of the A-alcohol metabolic target test is the same as the anti-aging target test method of Test Example 1. The difference is that the mRNA used is different. The mRNA of the A-alcohol metabolic target is RARA, RARB, RARG, RXRA, RXRB, RXRG, CRBP-I.

[0081] A-alcohol metabolism targets RARA mRNA, RARB mRNA, RARG mRNA, RXRA mRNA, The test results of RXRB mRNA, RXRG mRNA, and CRBP-I RNA expression are shown in Table 2.

[0082]

[0083] From Table 2 and Figure 5-11 The data show that when Gravi-A and 111 are combined in different ratios, they can promote the expression of different retinol metabolic targets. When the mass ratio of Gravi-A / 111 is 9:1 (mild anti-aging composition 1), it has a synergistic effect on promoting the expression of RXRB; when the mass ratio of Gravi-A / 111 is 5:5 (mild anti-aging composition 3), it has a synergistic effect on promoting the expression of RARG. Of particular note, when the mass ratio of Gravi-A / 111 is 2:8 (mild anti-aging composition 4), it has a significant synergistic effect on the mRNA expression of the retinol metabolism targets RARA, RARB, RARG, RXRA, RXRB, RXRG, and CRBP-I. Therefore, the composition of the present invention can increase collagen synthesis and reduce collagen degradation, thereby reducing wrinkles and improving skin elasticity.

[0084] Test Example 3 Inflammation Target Test The inflammatory target tested in the present invention is IL-1α.

[0085] The experimental process of the inflammatory target test is the same as the anti-aging target test method of Test Example 1. The difference is that the RNA used is different. The RNA of the inflammatory target is IL-1α.

[0086] The test results of the inflammatory target IL-1α expression are shown in Table 3.

[0087]

[0088] From Table 3 and Figure 12 The data shows that only at a Gravi-A / 111 mass ratio of 2:8 (mild anti-aging composition 4) did expression of the pro-inflammatory cytokine IL-1a significantly decrease. These results demonstrate that the composition of the present invention, without the addition of anti-irritant components, can effectively reduce the expression of molecular targets of inflammatory responses, thereby reducing skin irritation.

[0089] Test Example 4 Cytotoxicity Test Human fibroblasts in the exponential growth phase were prepared into single cell suspensions and cultured at a concentration of 3 x 10 3Cells were seeded into 96-well plates at a density of 100 μL / well and cultured at 37°C for 24 h in a medium containing 10% fetal bovine serum to allow the cells to adhere to the wall. The final culture volume was 100 μL. The supernatant was then discarded and freshly prepared serum-free DMSO solutions of different concentrations of hydroxypinacolone 9-cis retinoate (111), hydroxypinacolone retinoate (HPR), and retinyl propionate (RP) were added. After incubation for 24 h, 10 μL of CCK-8 was added to each well. After gentle shaking to mix, the cells were continued to be cultured at 37°C. After 2 h, the 96-well culture plate was removed and the absorbance of each well was measured at a wavelength of 450 nm to observe the effects of 111, HPR, and RP on cell growth.

[0090] The results of the effects of hydroxypinacolone 9-cis retinoate (111), hydroxypinacolone retinoate (HPR), and retinol propionate (RP) on the cell viability of human fibroblasts are shown in the attached figure. Figure 13-15 shown.

[0091] The test results show that 111, HPR and RP have no significant effect on the survival rate of fibroblasts at concentrations of 0.001-10 ppm. It is well known in the art that when the components do not undergo chemical reactions, if the components are not cytotoxic individually, then the combination of the components will not be cytotoxic either. Since 111, HPR and RP of the present invention are all retinol derivatives and do not react with each other, the mild anti-aging composition of the present invention is also not cytotoxic.

[0092] Example 3 Preparation of Cosmetic Mild Anti-aging Composition According to the components and contents in Table 4, an essence lotion containing a mild anti-aging composition was prepared.

[0093]

[0094] The preparation method of cosmetics comprises the following steps: Step 1: Mix all ingredients in Phase A and heat to 80°C until completely dissolved. Stir and mix until uniform. Step 2: Combine all ingredients in Phase B and heat to 80°C until completely dissolved. Stir and mix until uniform. Step 3: Add phase A to phase B at 80°C and homogenize at 4000 rpm for 3 min until uniform; Step 4: Cool the ingredients in step 3 to 60°C and add phase C, stirring well. Step 5: Continue to cool down to below 50°C, add phase D and phase E in sequence, and stir evenly until the temperature reaches below 40°C to obtain the essence lotion.

[0095] The prepared cosmetics include but are not limited to lotion, lotion, cream, mask, gel and essence, and can be prepared into different types of cosmetics as needed.

[0096] Test Example 5: Human anti-aging evaluation experiment of the mild anti-aging composition used in cosmetics.

[0097] To evaluate the anti-wrinkle, firming, and moisturizing effects of the essence lotion prepared in Example 3, 38 healthy women aged 38-60 years who reported experiencing wrinkles, fine lines, and sagging on their face and neck were selected. Exclusion criteria included those with obvious scars or birthmarks in the test area that could affect test results; those with self-reported severely sensitive skin or severely acne-prone skin; those with severe allergies; those who were pregnant, breastfeeding, or planning to become pregnant during the test period; those with severe heart, liver, or kidney damage or severe immunosuppression; those with mental illness, severe endocrine disease, or those taking oral contraceptives; those who were chronically dependent on hormones or antihistamines for treatment or maintenance; those who had participated in drug clinical trials or other trials within the past 60 days; those who had used medications that could affect test results within the past two weeks; those who had received medical care, beauty salon care, or treatment for their facial skin within the past 90 days; and those who had taken or applied topical cosmetics or health supplements with similar efficacy that could affect test results within the past 30 days. During the trial, the subjects agreed not to use any cosmetics, drugs or health products that might affect the results. Finally, 38 people participated in the test, of whom 3 withdrew from the trial for personal reasons. The effective sample size was 35. The subjects had Asian skin type.

[0098] The results are as attached Figure 16-28 As shown, compared with before use, after using the essence lotion of Example 3 for 7, 14 and 28 days, the length of forehead wrinkles was significantly reduced, the length of glabellar wrinkles was significantly reduced, the length of crow's feet wrinkles was significantly reduced, the length of nasolabial folds was significantly reduced, the length of neck wrinkles was significantly reduced, the length of mouth corner wrinkles was significantly reduced, the skin glossiness was significantly improved, the number of pores was significantly reduced, the wrinkles on the cheek skin were significantly improved, the moisture content of the skin stratum corneum was significantly increased, and the skin firmness parameter F4 was significantly reduced (the smaller the F4 value, the higher the skin firmness and the better the recovery ability). Strong. Conversely, the larger the F4 value, the lower the skin's firmness and the weaker its recovery ability. Therefore, the F4 value can be used as an important indicator for assessing skin firmness and aging). Skin elasticity parameters R2 and R5 have significantly improved (R2 is considered the parameter closest to the true state of skin elasticity. A larger R2 value indicates that the skin can recover to its original state faster after being subjected to external forces, showing better elasticity; a larger R5 value indicates that the skin's ability to recover after elastic deformation is stronger, that is, the skin can maintain good retraction properties after being subjected to external forces, which is also a key manifestation of skin elasticity).

[0099] In summary, from the above experimental results, it can be seen that through experimental testing of its anti-aging targets (type I collagen, type III collagen, type IV collagen and elastin) and retinol metabolic targets (RAR, RXR and CRBPI) at the mRNA gene expression level, the mild anti-aging composition of the present invention, the mass ratio of hydroxypinacolone 9-cis retinoic acid ester and Gravi-A is (1~8): (2~9). Within this ratio range, it can have a synergistic anti-aging effect; especially when the mass ratio of hydroxypinacolone 9-cis retinoic acid ester and Gravi-A in the composition is 8:2, through the inflammatory target IL-1α test, the composition of the present application can effectively reduce the expression of molecular targets of inflammatory response without adding anti-irritation components, thereby reducing skin irritation.

[0100] The above is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, etc. made within the spirit and principles of the present invention shall be included in the scope of protection of the present invention. For those skilled in the art, various modifications and improvements can be made without departing from the concept of the present invention, and these shall fall within the scope of protection of the present invention. Therefore, the scope of protection of the patent for this invention shall be based on the appended claims.

Claims

1. A mild anti-aging composition, characterized in that: The invention comprises hydroxypinacolone 9-cis retinoic acid ester and Gravi-A, wherein Gravi-A is composed of hydroxypinacolone retinoic acid ester and retinol propionate in a mass ratio of 5:

9.

2. A mild anti-aging composition according to claim 1, characterized in that: The mass ratio of the hydroxypinacolone 9-cis retinoic acid ester to Gravi-A is (1-8): (2-9).

3. A mild anti-aging composition according to claim 2, characterized in that: The mass ratio of hydroxypinacolone 9-cis retinoic acid ester to Gravi-A is (1-5): (5-9).

4. A mild anti-aging composition according to claim 2, characterized in that: The mass ratio of the hydroxypinacolone 9-cis retinoic acid ester to Gravi-A is 8:

2.

5. A method for preparing the mild anti-aging composition according to any one of claims 1 to 4, characterized in that: The components of the composition are mixed in proportion, a solvent is added, and the mixture is heated and stirred to obtain a mild anti-aging composition.

6. A cosmetic comprising the mild anti-aging composition according to any one of claims 1 to 4.

7. A mild anti-aging cosmetic composition according to claim 6, characterized in that: The cosmetics include any one of water, milk, cream, facial mask, gel, essence and essence milk.

8. The mild anti-aging composition cosmetic according to claim 6, characterized in that: The mass percentage of the mild anti-aging composition in the cosmetic is 0.01-0.3%.

9. Use of the mild anti-aging composition according to any one of claims 1 to 4 in the preparation of cosmetics.

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