A mild anti-aging composition and its preparation method, cosmetics and application

Abstract

The present invention provides a mild anti-aging composition and a preparation method thereof, cosmetics and applications, and belongs to the field of cosmetic technology. The mild anti-aging composition of the present invention includes 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. When the composition of the present invention is within a specific 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, without adding anti-irritation components, the expression of molecular targets of inflammatory response can be effectively reduced to reduce the irritation of the skin. While achieving excellent anti-aging effects, this composition also takes into account the mild needs of the skin, reduces the amount of hydroxypinacolone 9-cis retinoic acid ester, reduces costs, and has broad prospects for applying it to the 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 and a preparation method thereof, cosmetics and applications. Background Art

[0002] As an early indicator of systemic aging, skin aging has always been valued by researchers in the field of aging. In addition to the inevitable natural aging process, how to effectively delay skin aging has always been a concern of dermatologists and skin care product R&D experts. Early skin aging manifests itself in sagging skin, wrinkles, dry skin, and dull complexion. As we age, the elasticity and collagen of the skin decrease, resulting in fine lines on the face, especially around the eyes, forehead, and corners of the mouth, which are areas with rich expressions. Skin aging may also lead to uneven pigmentation, spots or freckles, making the skin look darker.

[0003] As early as the 1980s, studies have confirmed that vitamin A acid can improve skin aging. In the in-depth research of anti-aging products, many studies have revealed the positive effects of topical use of vitamin A acid in reducing aging damage; however, the irritation reactions that vitamin A acid may cause, such as erythema, stinging or burning, limit its application in a wider range of fields. Therefore, in the research of anti-aging products, researchers have developed a variety of vitamin A acid derivatives, such as retinol, retinal, retinoic acid, etc., among which retinoic acid is considered to be one of the effective substances in slowing down 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 group of retinoic acid and can directly act on retinoic acid receptors to exert its effect without being converted into retinoic acid, thus performing well in anti-aging, inhibiting sebum secretion, whitening and lightening spots. As a new generation of vitamin A acid derivatives, HPR has advantages over previous derivatives in terms of stability, safety and effectiveness. Although HPR is significantly less irritating than retinol, the amount of HPR added in cosmetics is subject to certain restrictions. Excessive addition may increase the irritation of the product and also affect the stability of the product.

[0005] Hydroxypinacolone 9-cis retinoic acid ester, whose chemical structure is isomerized relative to that of HPR, CN116947723 A discloses hydroxypinacolone 9-cis retinoic acid ester, its preparation method and application, in which hydroxypinacolone 9-cis retinoic acid ester shows significantly lower irritation and significantly improved skin anti-aging effect than HPR. Adding hydroxypinacolone 9-cis retinoic acid ester to cosmetics can achieve the purpose of reducing the irritation of retinoic acid ester substances to the skin and enhancing its anti-aging efficacy. However, due to the high price of hydroxypinacolone 9-cis retinoic acid ester, if it is added to cosmetics alone, it will lead to a significant increase in cost, thus lacking economic benefits.

[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 gentle 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 and a preparation method, cosmetics and applications thereof.

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

[0009] On the basis of a large number of experimental studies, the research and development team of the present invention combined 111 and Gravi-A, and tested the expression 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 level. It was found that 111 and Gravi-A in the composition have synergistic promotion of gene expression within a specific ratio range to achieve the anti-aging effect; at the same time, by testing the inflammatory target IL-1α, the research and development team of the present invention unexpectedly found that the composition of the present application can effectively reduce the expression of the molecular target of the inflammatory response without adding anti-irritation components, thereby reducing the irritation of the skin. Therefore, the mild anti-aging composition of the present invention not only achieves excellent anti-aging effects, but also takes into account the mild needs of the skin, and reduces the amount of hydroxypinacolone 9-cis retinoic acid ester, reducing costs, and has broad prospects for its application in the preparation of cosmetics.

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

[0011] The patent CN 115887280 A applied by the research and development team of the present invention discloses that hydroxypinacolone retinoic acid ester (HPR) and retinol propionate (RP) are combined in a mass ratio of (5-40):9 to achieve a synergistic effect. This synergistic effect is particularly prominent when hydroxypinacolone retinoic acid ester and retinol propionate are combined in a mass ratio of 5:9, which can significantly increase the expression of important series of proteins related to aging, Col-IV, CRBPI, RARB, and Elastin. Among them, the expression of type IV collagen is increased by more than 4 times, the expression of CRBP-1 is increased by more than 30 times, the expression of RARB is increased by more than 5 times, and the expression of Elastin is increased by more than 300 times, and there is no 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 metabolic targets (RAR, RXR and CRBPI) at the mRNA level, thereby achieving the anti-aging effect on the skin.

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

[0015] The 111 and Gravi-A of the present invention can not only synergistically increase the expression of an important series of anti-aging related proteins within the above-mentioned mass ratio range, but also reduce the amount of expensive hydroxypinacolone 9-cis retinoic acid ester, so that the amount of hydroxypinacolone 9-cis retinoic acid ester is reduced by at least 50%. Exemplarily, the mass ratio of 111 to Gravi-A is 1:9; or 2:8; or 5:5, and the embodiments of the present invention provide the relevant content 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 is at the above mass ratio, it can not only promote the expression of the anti-aging target collagen, but also promote the expression of the target of retinol metabolism, and effectively reduce the expression of the molecular target of inflammatory response, thus well balancing the excellent anti-aging effect and excellent mildness. The embodiments of the present invention provide the components of the mild anti-aging composition with the relevant contents.

[0018] On the other hand, the present invention also provides a method for preparing a mild anti-aging composition. 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 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, vaseline, 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≥10, and preferably 10-24, more preferably 12-20, such as 14, 16, 17, 18. The Cn fatty acids are such as 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 alcohol. The Cn fatty acid esters and Cn fatty alcohol esters include isopropyl myristate, myristyl myristate, myristyl lactate, hexadecyl lactate, isopropyl palmitate, isooctyl palmitate, butyl stearate, isooctyl stearate, monoglyceride stearate, polyethylene glycol stearate, decyl oleate, glyceryl caprylate, glyceryl caprate, and glyceryl trioleate.

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

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

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

[0025] Exemplarily, the moisturizer includes one or more of propylene glycol, 1,3-propylene glycol, methyl propanediol, glycerol, diglycerol, 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 moisturizer in the cosmetic is in the range of 0.1 to 10%.

[0026] Exemplarily, the emulsifier includes glyceryl stearate, PEG-100 stearate, cetearyl alcohol, ceteareth-20, PEG-40 stearate, cetearyl glucoside, C14-22 alcohol, C12-20 alkyl glucoside, polyglyceryl-3 methyl glucose 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 olive oil ester, sorbitan olive oil ester, sucrose polystearate, cetyl palmitate, olive oil PEG-6 polyglyceryl-6 esters, lecithin, hydrogenated lecithin, and one or more thereof. The mass percentage of the emulsifier in the cosmetic is in the range of 0.1 to 5%.

[0027] Exemplarily, the thickener includes one or more of hydroxyethyl cellulose, hydroxypropyl methylcellulose, xanthan gum, sodium hydroxypropyl starch phosphate, acrylic acid (esters) / C10-30 alkyl acrylate cross-linked polymer, acrylic acid (esters) copolymer, acrylic acid (esters) copolymer sodium, carbomer, ammonium acryloyldimethyl taurate / VP copolymer, and sodium polyacryloyldimethyl taurate. The mass percentage of the thickener in the cosmetic is 0.1-1%.

[0028] Exemplarily, 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] Exemplarily, the chelating agent includes one or more of disodium ethylenediaminetetraacetate, sodium phytate, tetrasodium glutamate diacetate, disodium dihydrogen pyrophosphate, sodium citrate, and trisodium ethylenediamine disuccinate. 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 mass percentage content of the pH regulator in the cosmetic is in the range of 0.1-1%.

[0031] Exemplarily, the other active ingredients include one or more of tocopherol (vitamin E), tocopherol acetate, ectoine, niacinamide, etc. The mass percentage of other active ingredients in the cosmetics 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 practical applications, 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 beneficial effects: the mild anti-aging composition of the present invention has a synergistic anti-aging effect within a specific ratio range, and the composition can effectively reduce the expression of molecular targets of inflammatory response without adding anti-irritation components, thereby reducing skin irritation. At the same time, the amount of hydroxypinacolone 9-cis retinoic acid ester is reduced, the production cost is reduced, and the application of the composition in the preparation of cosmetics has broad prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0035] The drawings constituting a part of the present application are used to provide a further understanding of the present invention. The exemplary embodiments of the present invention and their descriptions are used to explain the present invention and do not constitute an improper limitation of the present invention. In the drawings:

[0036] Figure 1 This is a graph showing the effect of different proportions of mild anti-aging compositions on Col I mRNA expression in the anti-aging target test of Test Example 1;

[0037] Figure 2 This is a graph showing the effect of different proportions of mild anti-aging compositions on Col Ⅲ mRNA expression in the anti-aging target test of Test Example 1;

[0038] Figure 3 This is a graph showing the effect of different proportions of mild anti-aging compositions on Col IV mRNA expression in the anti-aging target test of Test Example 1;

[0039] Figure 4 This is a graph showing the effect of different proportions of mild anti-aging compositions on Elastin mRNA expression in the anti-aging target test of Test Example 1;

[0040] Figure 5 This is a graph showing the effect of different proportions of mild anti-aging compositions on RARA mRNA expression in the A-alcohol metabolism target test in Test Example 2;

[0041] Figure 6This is a graph showing the effect of different proportions of mild anti-aging compositions on RARB mRNA expression in the A-alcohol metabolism target test in Test Example 2;

[0042] Figure 7 This is a graph showing the effect of different proportions of mild anti-aging compositions on RARG mRNA expression in the A-alcohol metabolism target test in Test Example 2;

[0043] Figure 8 This is a graph showing the effect of different proportions of mild anti-aging compositions on RXRA mRNA expression in the A-alcohol metabolism target test in Test Example 2;

[0044] Fig. 9 This is a graph showing the effect of different proportions of mild anti-aging compositions on RXRB mRNA expression in the A-alcohol metabolism target test in Test Example 2;

[0045] Fig.10 This is a graph showing the effect of different proportions of mild anti-aging compositions on RXRG mRNA expression in the A-alcohol metabolism target test in Test Example 2;

[0046] Fig.11 This is a graph showing the effect of different proportions of mild anti-aging compositions on CRBP-I mRNA expression in the A-alcohol metabolism target test in Test Example 2;

[0047] Fig.12 This is a graph showing the effect of different proportions of mild anti-aging compositions on IL-1α expression in the inflammatory target test of Test Example 3;

[0048] Fig.13 This is a graph showing the effect of hydroxypinacolone 9-cis retinoic acid ester on the survival rate of human fibroblasts in the cytotoxicity test of Test Example 4;

[0049] Fig.14 This is a graph showing the effect of hydroxypinacolone retinoic acid ester on the survival rate of human fibroblasts in the cytotoxicity test of Test Example 4;

[0050] Fig.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;

[0051] Fig.16 This is the result of the evaluation of the length of forehead wrinkles by the essence lotion of Example 3 in the human anti-aging evaluation of Test Example 5;

[0052] Fig.17 This is a graph showing the evaluation results of the length of the glabellar lines by the essence lotion of Example 3 in the human anti-aging evaluation of Test Example 5;

[0053] Fig.18 This is a graph showing the evaluation result of the essence lotion of Example 3 on the length of crow's feet in the human anti-aging evaluation of Test Example 5;

[0054] Fig.19 This is a graph showing the evaluation results of the essence lotion in Example 3 on the length of nasolabial folds in the human anti-aging evaluation in Test Example 5;

[0055] Fig. 20 This is a graph showing the evaluation results of the essence lotion of Example 3 on the length of neck wrinkles in the human anti-aging evaluation of Test Example 5;

[0056] Fig.21 This is the evaluation result of the length of corner lines of the mouth by the essence lotion of Example 3 in the human anti-aging evaluation of Test Example 5;

[0057] Fig. 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;

[0058] Fig.23 This is the result diagram of the evaluation of the number of pores by the essence lotion of Example 3 in the human anti-aging evaluation of Test Example 5;

[0059] Fig.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;

[0060] Fig.25 This is a graph showing the evaluation results of the moisture content of the stratum corneum of the skin by the essence lotion of Example 3 in the human anti-aging evaluation of Test Example 5;

[0061] Fig.26 This is a graph showing the evaluation results of the skin firmness parameter F4 of the essence lotion of Example 3 in the human anti-aging evaluation of Test Example 5;

[0062] Fig. 27 This is a graph showing the evaluation results of the skin elasticity parameter R2 of the essence lotion of Example 3 in the human anti-aging evaluation of Test Example 5;

[0063] Fig.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

[0064] In order to clearly and completely describe the technical solutions in the embodiments of the present invention below, it is obvious that the described embodiments are only part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0065] In order to facilitate the understanding of the present invention, the present invention will be described more comprehensively below, and preferred embodiments of the present invention are given. However, it should be understood that these embodiments are only used for more detailed description and should not be understood as limiting the present invention in any form, that is, not limiting the scope of protection of the present invention.

[0066] The present invention will be further described below in conjunction with the embodiments.

[0067] The various components of the mild anti-aging composition of the present invention are described in detail.

[0068] Hydroxypinacolone 9-cis retinoic acid ester

[0069] Hydroxypinacolone 9-cis retinoic acid ester, CN 116947723 A discloses the preparation method and application of the substance. The patent tests the expression levels of aging-related genes and RAR / RXR receptor expression levels, proving that hydroxypinacolone 9-cis retinoic acid ester has lower irritation and improved skin anti-aging effect than HPR.

[0070] The mild anti-aging composition of the present invention comprises hydroxypinacolone 9-cis retinoic acid ester.

[0071] The structure of hydroxypinacolone 9-cis retinoic acid ester is as follows:

[0072]

[0073] The hydroxypinacolone 9-cis retinoic acid ester used in the present invention is prepared by the preparation method of CN 116947723 A, and the specific method is as follows:

[0074] (1) dissolving 9-cis retinoic acid in a dimethylformamide solution, and adding cesium carbonate and 1-chloropinacolone in sequence;

[0075] (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;

[0076] (3) combining the organic extracts, drying them, and concentrating them under reduced pressure;

[0077] (4) Purify by silica gel column chromatography using a 5:1 volume ratio of hexane / ethyl acetate mixture.

[0078] Hydroxypinacolone retinoate

[0079] Hydroxypinacolone retinoic acid ester is a new type of retinoic acid derivative formed by the dehydration condensation of retinoic acid and pinacol. It retains the functional group of retinoic acid and can directly act on retinoic acid receptors to exert its effect without converting into retinoic acid, thus performing well in anti-aging, inhibiting sebum secretion, whitening and lightening spots, etc.

[0080] The mild anti-aging composition of the present invention comprises hydroxypinacolone retinoic acid ester.

[0081] The structure of hydroxypinacolone retinoic acid ester is as follows:

[0082]

[0083] Hydroxypinacolone retinoic acid ester can be used without particular limitation as long as it is, for example, hydroxypinacolone retinoic acid ester commonly used in cosmetics.

[0084] As hydroxypinacolone retinoic acid ester, a commercially available product can be used.

[0085] Retinyl Propionate

[0086] The mild anti-aging composition of the present invention comprises retinol propionate.

[0087] The structure of retinol propionate is as follows:

[0088]

[0089] Retinol propionate can be used without particular limitation as long as it is, for example, a retinol propionate commonly used in cosmetics.

[0090] As retinol propionate, a commercially available product can be used.

[0091] 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:

[0092] 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 instruments, kits, cells and reagents in the art. The operating procedures of the instruments and kits are carried out in accordance with the instructions of their instructions.

[0093] Anti-aging target type I collagen

[0094] 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 and maintains the skin's stability and elasticity.

[0095] Anti-aging target: type III collagen

[0096] Type III collagen (Col III) exists in the skin in the form of fine fibers, intertwined with type I collagen, providing suppleness and elasticity to the skin. It helps maintain skin flexibility and reduce the formation of wrinkles.

[0097] Anti-aging target type IV collagen

[0098] 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 meshwork of the BM, an important connecting layer between the epidermis and dermis of the skin, and is essential for maintaining the integrity and function of the skin.

[0099] Anti-aging target elastin

[0100] Elastin, together with collagen fibers, constitutes the extracellular matrix of the skin, providing structural support for the skin. It forms a complex network in the dermis of the skin, interacting with collagen fibers to maintain the shape and stability of the skin. The maintenance of elastin is very important for anti-aging and keeping the skin youthful.

[0101] Alanine metabolism targets RAR and RXR

[0102] Retinol is first converted into retinoic acid (RA) in the body, and this process is a key step for retinol to exert its biological activity. Retinoic acid then binds to the retinoic acid receptors (RAR and RXR) in the cell to form a dimer complex, which then binds to the retinoic acid response element (RARE or RXRE) to regulate the activation and expression of mRNA in the cell nucleus. After retinol binds to RAR and RXR, it can promote the maturation and differentiation of skin cells, which is very important for maintaining the normal renewal and repair process of the skin. At the same time, the activation of RAR and RXR can increase the synthesis of collagen and reduce the degradation of collagen, thereby reducing wrinkles and improving skin elasticity.

[0103] ALCOHOL METABOLIZATION TARGET CRBPI

[0104] CRBPI indirectly regulates the metabolic rate and direction of A-alcohol in cells through its ability to bind A-alcohol. By binding to A-alcohol, CRBPI can affect the conversion of A-alcohol to retinoic acid (RA), thereby regulating the concentration and activity of RA in cells.

[0105] Inflammatory target IL-1α

[0106] IL-1α is a cytokine with strong inflammatory and immune amplification effects, which is mainly expressed by various cell types in response to external stimuli. Alcohol components often promote the expression of IL-1α, and the composition of the present invention can reduce the expression of IL-1α.

[0107] Example 1 Preparation of composition

[0108] Mild anti-aging composition 1: (Gravi-A / 111=9:1)

[0109] 1 g of hydroxypinacolone 9-cis retinoic acid ester, 3.214 g of hydroxypinacolone retinoic acid ester and 5.786 g of retinol propionate were mixed in proportion, 190 g of dimethyl sulfoxide was added, the mixture was heated to not more than 80° C. and stirred evenly, and the mixture was cooled to obtain a mild anti-aging composition 1.

[0110] Mild anti-aging composition 2: (Gravi-A / 111=8:2)

[0111] 2 g of hydroxypinacolone 9-cis retinoic acid ester, 2.857 g of hydroxypinacolone retinoic acid ester and 5.143 g of retinol propionate were mixed in proportion, 190 g of dimethyl sulfoxide was added, the mixture was heated to not more than 80° C. and stirred evenly, and the mixture was cooled to obtain a mild anti-aging composition 2.

[0112] Mild anti-aging composition 3: (Gravi-A / 111=5:5)

[0113] 5 g of hydroxypinacolone 9-cis retinoic acid ester, 1.786 g of hydroxypinacolone retinoic acid ester and 3.214 g of retinol propionate were mixed in proportion, 190 g of dimethyl sulfoxide was added, the mixture was heated to not more than 80° C. and stirred evenly, and the mixture was cooled to obtain a mild anti-aging composition 3.

[0114] Mild anti-aging composition 4: (Gravi-A / 111=2:8)

[0115] 8 g of hydroxypinacolone 9-cis retinoic acid ester, 0.714 g of hydroxypinacolone retinoic acid ester and 1.286 g of retinol propionate were mixed in proportion, 190 g of dimethyl sulfoxide was added, the mixture was heated to not more than 80° C. and stirred evenly, and the mixture was cooled to obtain a mild anti-aging composition 4.

[0116] Mild anti-aging composition 5: (Gravi-A / 111=4:6)

[0117] 6 g of hydroxypinacolone 9-cis retinoic acid ester, 1.429 g of hydroxypinacolone retinoic acid ester and 2.571 g of retinol propionate were mixed in proportion, 190 g of dimethyl sulfoxide was added, the mixture was heated to not more than 80° C. and stirred evenly, and the mixture was cooled to obtain a mild anti-aging composition 5.

[0118] Mild anti-aging composition 6: (Gravi-A / 111=3:7)

[0119] 7 g of hydroxypinacolone 9-cis retinoic acid ester, 1.071 g of hydroxypinacolone retinoic acid ester and 1.929 g of retinol propionate were mixed in proportion, 190 g of dimethyl sulfoxide was added, the mixture was heated to not more than 80° C. and stirred evenly, and the mixture was cooled to obtain a mild anti-aging composition 6.

[0120] Example 2 Preparation of Comparative Example

[0121] Comparative Example 1: (Gravi-A / 111=1:9)

[0122] 9 g of hydroxypinacolone 9-cis retinoic acid ester, 0.357 g of hydroxypinacolone retinoic acid ester 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 temperature was lowered to obtain Comparative Example 1.

[0123] Comparative Example 2: (Gravi-A)

[0124] 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, the mixture was heated to no more than 80° C. and stirred evenly, and the mixture was cooled to obtain Comparative Example 2.

[0125] Comparative Example 3: (111)

[0126] 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.

[0127] Test Example 1 Anti-aging target test

[0128] 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 (Elastin).

[0129] 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 5 The cells were inoculated on a 6-well plate at a density of cells / mL, with a final culture volume of 2 mL, and cultured at 37°C with a medium containing 10% fetal bovine serum until the cells adhered to the wall. Primary fibroblasts were inoculated into a low-adsorption culture plate and cultured at 37°C with a medium containing 10% fetal bovine serum for 3 to 5 days to form spheroids. After the cells adhered to the wall or spheroids were formed, the supernatant was discarded, and a freshly prepared composition of Example 1 and Example 2 (serum-free) at a concentration of 0.075 μg / mL 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 RNA extraction standard operating procedure, and reverse transcribed into cDNA for qPCR detection of key gene expression, and the remaining RNA was packaged and stored.

[0130] 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 routinely used by those skilled in the art, and each operation step was performed with reference to the kit instructions.

[0131] mRNA expression detection method for Col I, Col III, Col IV and Elastin: Total RNA was extracted from cell samples using the Trizol RNA extraction kit, and the extracted RNA was reverse transcribed into cDNA using reverse transcriptase. The cDNA of the target gene was amplified using specific primers, and the amplified product was monitored in real time by adding fluorescent dye. The intensity of the fluorescent signal is proportional to the mRNA level of the target gene. The internal reference gene GAPDH was used for standardization to calculate the relative expression.

[0132] 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 among groups. "*" indicates p < 0.05, indicating that there is a significant difference compared with the Control, Gravi-A and 111 groups.

[0133] 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.

[0134] Table 1 Anti-aging target test results

[0135]

[0136] From Table 1 and Figure 1-4 The 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 to 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 to 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 to only Gravi-A or 111.

[0137] 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 lower 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).

[0138] Test Example 2 A-alcohol metabolism target test

[0139] 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.

[0140] The experimental process of the ALPHA 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 ALPHA metabolic target is RARA, RARB, RARG, RXRA, RXRB, RXRG, and CRBP-I.

[0141] The test results of the expression of ALCOHOL metabolic targets RARA mRNA, RARB mRNA, RARG mRNA, RXRA mRNA, RXRB mRNA, RXRGmRNA, and CRBP-I RNA are shown in Table 2.

[0142] Table 2 Results of A-alcohol metabolic target test

[0143]

[0144] From Table 2 and Figure 5-11 The data show that when Gravi-A and 111 are in different compounding 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 the effect of synergistically promoting the expression of RXRB; when the mass ratio of Gravi-A / 111 is 5:5 (mild anti-aging composition 3), it has the effect of synergistically promoting the expression of RARG; it is particularly noteworthy that when the mass ratio of Gravi-A / 111 is 2:8 (mild anti-aging composition 4), the mRNA expression of the retinol metabolic targets RARA, RARB, RARG, RXRA, RXRB, RXRG and CRBP-I all have significant synergistic effects. Therefore, the composition of the present invention can increase the synthesis of collagen and reduce the degradation of collagen, thereby reducing wrinkles and improving skin elasticity.

[0145] Test Example 3 Inflammation Target Test

[0146] The inflammatory target tested in the present invention is IL-1α.

[0147] The experimental process of the inflammatory target test is the same as the anti-aging target test method of Test Example 1, except that the RNA used is different, and the RNA of the inflammatory target is IL-1α.

[0148] The test results of the inflammatory target IL-1α expression are shown in Table 3.

[0149] Table 3 Inflammatory target test results

[0150]

[0151] From Table 3 and Fig.12 The data show that only at a mass ratio of Gravi-A / 111 of 2:8 (mild anti-aging composition 4), the expression of the pro-inflammatory cytokine IL-1a was significantly reduced. The above results show that the composition of the present invention can effectively reduce the expression of molecular targets of inflammatory response and thus reduce skin irritation without adding anti-irritant components.

[0152] Test Example 4 Cytotoxicity Test

[0153] Human fibroblasts in the exponential growth phase were prepared into single cell suspensions and cultured at 3x10 3 Cells were inoculated at a density of 100 μL / well on a 96-well plate and cultured at 37°C for 24 hours with 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 retinoic acid ester (111), hydroxypinacolone retinoic acid ester (HPR), and retinol propionate (RP) were added. After incubation for 24 hours, 10 μL of CCK-8 was added to each well, and after slight shaking to mix, the cells were continued to be cultured at 37°C. After 2 hours, the 96-well culture plate was taken out and the absorbance OD of each well was measured at a wavelength of 450 nm to observe the effects of 111, HPR, and RP on cell growth.

[0154] The results of the effects of hydroxypinacolone 9-cis retinoic acid ester (111), hydroxypinacolone retinoic acid ester (HPR), and retinol propionate (RP) on the cell viability of human fibroblasts are shown in the attached figure. Figure 13-15 shown.

[0155] It can be seen from the test results that 111, HPR and RP have no significant effect on the survival rate of fibroblasts at a concentration of 0.001-10 ppm. It is well known in the art that when the components do not undergo chemical reactions and are not cytotoxic individually, 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.

[0156] Example 3 Preparation of cosmetics of mild anti-aging composition

[0157] According to the components and contents in Table 4, an essence lotion containing a mild anti-aging composition was prepared.

[0158] Table 4 Cosmetic ratio table

[0159]

[0160] The preparation method of cosmetics comprises the following steps:

[0161] Step 1: Mix all ingredients of phase A and heat to 80°C until completely dissolved, stirring and mixing until uniform;

[0162] Step 2: Mix all ingredients of phase B and heat to 80°C until completely dissolved, stirring and mixing until uniform;

[0163] Step 3: Add phase A to phase B at 80°C and homogenize at 4000 rpm for 3 min until uniform;

[0164] Step 4: Cool the ingredients in step 3 to 60°C and add phase C, stirring evenly;

[0165] 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.

[0166] The prepared cosmetics include but are not limited to water, milk, cream, mask, gel and essence, and can be prepared into different types of cosmetics as needed.

[0167] Test Example 5 Human anti-aging evaluation experiment of the mild anti-aging composition used in cosmetics.

[0168] In order to evaluate the anti-wrinkle, firming, moisturizing and other skin anti-aging effects of the essence milk prepared in Example 3, 38 healthy women aged 38-60 were selected, who reported that their faces and necks were accompanied by wrinkles or fine lines, and sagging. The exclusion criteria were those with obvious scars, birthmarks, etc. in the test area that affected the test results, those with severely sensitive skin (self-declared) and severely acne-prone skin, those with highly allergic constitutions, women who were pregnant, breastfeeding or intended to be pregnant during the test period, those with severe heart, liver, and kidney damage and severe immunodeficiency, those with mental illness, severe endocrine diseases, and oral contraceptives, those who relied on hormones, antihistamines for long-term treatment or maintenance of physical condition, those who participated in drug clinical trials or other trials within 60 days, or those who had used drugs that affected the test results within the past 2 weeks, those who had received medical care, beauty salon care or treatment for facial skin within 90 days, and those who started taking or applying cosmetics and health products of the same efficacy that may affect the test results within 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 which 3 withdrew from the trial for personal reasons. The effective sample size was 35. The subjects' skin type was Asian.

[0169] The results are attached Figure 16-28 As shown, compared with before use, after using the essence lotion of Example 3 for 7 days, 14 days 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 corners of mouth wrinkles was significantly reduced, the skin glossiness was significantly improved, the number of pores was significantly reduced, the cheek skin wrinkles were significantly improved, the moisture content of the skin stratum corneum was significantly improved, and the skin firmness parameter F4 was significantly reduced (the smaller the F4 value, the higher the skin firmness and the stronger the recovery ability). Strong. On the contrary, 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 to evaluate the firmness and aging degree of the skin), the skin elasticity parameters R2 and R5 have been significantly improved (the R2 value is considered to be the parameter value closest to the true state of skin elasticity. The larger the R2 value, the faster the skin can recover to its original state after being acted upon by external force, showing better elasticity; the larger the R5 value, the stronger the skin's recovery ability after elastic deformation, that is, the skin can maintain good retraction performance after being acted upon by external force, which is also one of the important manifestations of skin elasticity).

[0170] In summary, from the above experimental results, it can be seen that through experimental tests on its anti-aging targets (type I collagen, type III collagen, type IV collagen and elastin) and A-alcohol metabolic targets (RAR, RXR and CRBPI) at the mRNA gene expression level, the mass ratio of hydroxypinacolone 9-cis retinoic acid ester and Gravi-A of the mild anti-aging composition of the present invention is (1~8): (2~9). When it is 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.

[0171] The above is only a preferred embodiment of the invention, and is not intended to limit the invention. Any modifications, equivalent substitutions, etc. made within the spirit and principle of the invention shall be included in the protection scope of the invention. For ordinary technicians in this field, several modifications and improvements can be made without departing from the concept of the invention, which all belong to the protection scope of the invention. Therefore, the protection scope of the patent of the invention shall be based on the attached 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, and the mass ratio of hydroxypinacolone 9-cis retinoic acid ester and Gravi-A is 8:

2.

2. A method for preparing the mild anti-aging composition according to claim 1, 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.

3. A cosmetic comprising the mild anti-aging composition according to claim 1.

4. The cosmetic according to claim 3, characterized in that: The cosmetic is selected from any one of water, milk, cream, facial mask, gel and essence.

5. The cosmetic according to claim 3, characterized in that: The mass percentage of the mild anti-aging composition in the cosmetic is 0.01-0.3%.

6. Use of the mild anti-aging composition according to claim 1 in the preparation of anti-aging cosmetics.

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