Beauty composition and application thereof in whitening, anti-inflammatory and anti-aging products

Abstract

The invention provides a beautifying composition and application thereof in whitening, anti-inflammatory and anti-aging products, and belongs to the technical field of cosmetics. The beautifying composition is prepared from collagen peptide, sodium hyaluronate and ergothioneine, wherein the mass concentration ratio of the collagen peptide to the sodium hyaluronate to the ergothioneine is (75 to 85) to (3.5 to 10) to (0.4 to 3.5); the beautifying composition shows remarkable advantages in the aspects of collagen regeneration and skin improvement, the collagen content can be remarkably increased, and multi-effect collagen regeneration promotion is achieved by inhibiting collagen degrading enzyme MMP-1 and up-regulating TGF-beta1; the content of AQP3 and HA can be increased, and water locking and moisturizing of the skin are enhanced; by activating an Nrf2 pathway, the SOD2 level is improved, IL-6 and IL-1alpha are inhibited, and the antioxidant, anti-inflammatory and whitening effects are achieved; the thickness of an epidermal layer and the density of dermal fibroblasts can be remarkably improved, wrinkle resistance and repairing are effectively achieved, and the skin structure is improved.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of cosmetics, and particularly relates to a cosmetic composition and application thereof in whitening, anti-inflammatory and anti-aging products. BACKGROUND

[0002] Skin, as an important organ of the human body, bears the protection of environmental and mechanical damage, senses stimulation, regulates body temperature, and monitors immunity, and the like, multiple important physiological processes, but with age, the skin morphology and physiology gradually degenerate, and skin anti-aging products have been a hot spot in the cosmetics industry.

[0003] With natural aging, the epidermis first presents barrier damage, leading to a decrease in skin water content, a weakening of the proliferation ability of keratinocytes, a thinning of the epidermis, and the appearance of fine lines on the skin. Barrier damage leads to a decrease in the protective ability of the skin, and the skin is more susceptible to external damage, leading to the activation of MMPs, which degrade collagen and elastin in the dermis. At the same time, the fibroblast cytosol in the dermis presents aging due to reasons such as a decrease in mitochondrial oxygen supply and a shortening of telomeres in the genes in the cell nucleus, leading to a decrease in the proliferation ability of fibroblasts and a decrease in the ability of fibroblasts to secrete collagen and elastin, further exacerbating the decrease in extracellular matrix material in the dermis, and the supporting effect of the dermis on the skin decreases with age, and the skin gradually presents relaxation and sagging. In the problem of aging, due to the continuous contraction of the muscles, expression lines accumulate day by day, making the skin present an aged appearance, and expression lines are an important factor in the presentation of the aging state of the skin. The realization of the functions of the skin depends on the energy produced by respiration, and mitochondria produce a large amount of ROS while producing energy. Cell aging leads to a decrease in the ability to remove free radicals, and excessive ROS leads to a series of skin problems.

[0004] However, existing products have relatively single efficacy, and generally have the problem of poor whitening and anti-inflammatory effect. For example, Chinese Patent CN114903826A discloses an anti-aging composition, an anti-aging cosmetic, and a preparation method thereof. The anti-aging composition includes aminobutyric acid, hydrolyzed collagen, hydrolyzed sodium hyaluronate, centella asiatica extract, and chaenomeles fruit extract. The application verifies that the components have a good free radical scavenging effect through interaction in the implementation process, and can effectively reduce wrinkles, but the anti-inflammatory effect and whitening effect are not involved.

[0005] For example, Chinese patent CN110227050A discloses an anti-aging repair slow-release essence and an anti-aging repair mask prepared by using the same. The anti-aging repair mask liquid is composed of the following raw materials and mass percentages: glycerol 5%-10%, natural plant extract 5%-15%, butanediol 1%-1.5%, hydroxyethyl cellulose 2%-4%, sodium hyaluronate 2%-4%, lycine 1%-3%, polyacrylic acid 0.5%-1%, p-hydroxyacetophenone 0.2%-0.5%, citric acid 0.2%-0.5%, hydrogenated castor oil 0.5%-1%, and the balance is deionized water, and the total of the mass percentages of the raw materials is 100%. The anti-aging repair mask essence of the application has the effects of antioxidant, promoting cell metabolism, inhibiting the production of melanin, etc., can reduce skin wrinkles, skin problems such as pigmentation, and has strong functions of anti-aging and repair such as moisturizing, long-acting moisturizing, whitening, sunscreen, etc. The application has the effects of whitening and moisturizing, but the anti-inflammatory effect cannot meet the demand.

[0006] Therefore, it is necessary to develop a cosmetic composition with the effects of whitening, anti-inflammatory and anti-aging. SUMMARY

[0007] Based on the deficiencies of the prior art, the technical problem to be solved by the application is to provide a cosmetic composition and its application in whitening, anti-inflammatory and anti-aging products.

[0008] To this end, the application provides the following technical solutions: On the one hand, the application provides a cosmetic composition, which is composed of collagen peptide, sodium hyaluronate and ergothioneine, and the mass concentration ratio of the collagen peptide, the sodium hyaluronate and the ergothioneine is 75-85:3.5-10:0.4-3.5.

[0009] Preferably, the mass concentration ratio of the collagen peptide, the sodium hyaluronate and the ergothioneine is 76-84:3.7-8.5:0.6-3.3; More preferably, the mass concentration ratio of the collagen peptide, the sodium hyaluronate and the ergothioneine is 77-83:3.9-7.6:0.9-3.1; Further preferably, the mass concentration ratio of the collagen peptide, the sodium hyaluronate and the ergothioneine is 78-82:4.1-6.2:1.5-2.9; Still further preferably, the mass concentration ratio of the collagen peptide, the sodium hyaluronate and the ergothioneine is 79-81:4.3-5.0:2.0-2.7; Still further preferably, the mass concentration ratio of the collagen peptide, the sodium hyaluronate and the ergothioneine is 79-81:4.3-5.0:2.0-2.7; More preferably, the mass concentration ratio of the collagen peptide, sodium hyaluronate and ergothioneine is 80-81:4.7-4.9:2.0-2.3.

[0010] As some preferred embodiments, the mass concentration ratio of the collagen peptide, sodium hyaluronate and ergothioneine is selected from, but not limited to, 75:3.5:1.2, 75:4.0:2.0, 75:4.5:2.5, 75:5.0:3.0, 80:3.5:1.2, 80:4.0:2.0, 80.647:4.839:2.016, 80:5.0:2.5, 80:5.5:3.5, 85:3.5:1.2, 85:4.0:1.5, 85:4.5:2.0, 85:5.0:2.5 or 85:5.5:3.5.

[0011] As some preferred embodiments, the mass concentration ratio of the collagen peptide, sodium hyaluronate and ergothioneine is selected from, but not limited to, 75:3.5:1.2, 75:4.0:2.0, 75:4.5:2.5, 75:5.0:3.0, 80:3.5:1.2, 80:4.0:2.0, 80.647:4.839:2.016, 80:5.0:2.5, 80:5.5:3.5, 85:3.5:1.2, 85:4.0:1.5, 85:4.5:2.0, 85:5.0:2.5 or 85:5.5:3.5.

[0012] Specifically, the cosmetic composition further comprises a solvent selected from any one or more of water, polyhydric alcohol solvents, alcohol solvents, and oil solvents.

[0013] Preferably, the solvent is water.

[0014] Specifically, the collagen peptide comprises at least 20% of collagen tripeptide.

[0015] More preferably, the collagen peptide comprises 20%-100% of collagen tripeptide.

[0016] More preferably, the collagen peptide comprises 20%-30%, 30%-40%, 40%-50%, 50%-60%, 60%-70%, 70%-80%, 80%-90% or 90%-100% of collagen tripeptide.

[0017] Specifically, the ergothioneine is L-ergothioneine.

[0018] In another aspect, the present application provides a preparation method of the cosmetic composition of any one of the above, which comprises mixing the collagen peptide, sodium hyaluronate and ergothioneine in proportion, or dissolving the collagen peptide, sodium hyaluronate and ergothioneine in a solvent in proportion.

[0019] In yet another aspect, the present application provides a composition preparation comprising the cosmetic composition of any one of the above.

[0020] Specifically, the composition preparation further comprises any one or more of plant extracts, vitamin substances, bioactive peptide substances, enzyme substances, acid substances, polysaccharide substances or mineral elements.

[0021] Preferably, the plant extracts include, but are not limited to, any one or more of ginseng extract, grape seed extract, licorice extract, tea extract, aloe extract, witch hazel extract, rosemary extract, gotu kola extract, peony extract, angelica extract, coix seed extract, white hoelen extract, giant knotweed extract, mulberry bark extract, honeysuckle extract, orange peel extract, lemon extract, peach blossom extract, ginkgo leaf extract, medlar extract, rose extract, lavender extract, chamomile extract, atractylodes extract, peony root bark extract, scute extract, milkvetch extract, baizhi extract, yam extract, eucommia extract, notoginseng extract, salvia extract, chrysanthemum extract, cinnamon extract, magnolia bark extract, schisandra extract, amomum villosum extract, cistanche extract.

[0022] Specifically, the vitamin substances include fat-soluble vitamins and / or water-soluble vitamins.

[0023] Preferably, the fat-soluble vitamins include, but are not limited to, any one or more of vitamin A, vitamin D, vitamin E or vitamin K; Preferably, the water-soluble vitamins include, but are not limited to, any one or more of vitamin B1, vitamin B2, vitamin B3, vitamin B5, vitamin B6, vitamin B7, vitamin B12, vitamin C, choline or inositol.

[0024] Preferably, the bioactive peptide substances include, but are not limited to, any one or more of collagen peptides, elastin peptides, glutathione, casein phosphopeptides, hyaluronic acid hydrolyzed peptides, ceramide peptides, soybean peptides, corn peptides, sea cucumber peptides, oyster peptides, bird's nest peptides, camosine peptides or lactoferrin peptides.

[0025] Preferably, the enzyme substances include, but are not limited to, any one or more of superoxide dismutase, catalase, glutathione peroxidase, bromelain, papain, lysozyme.

[0026] Preferably, the acid substances include, but are not limited to, any one or more of vitamin A and its derivatives, vitamin C and its derivatives, fruit acid, salicylic acid, arbutin, theophylline acid, kojic acid, ferulic acid or azelaic acid.

[0027] In particular, the polysaccharide substance includes, but is not limited to, any one or more of fungal polysaccharides, animal polysaccharides, plant polysaccharides, and algal polysaccharides.

[0028] Preferably, the fungal polysaccharide includes, but is not limited to, any one or more of Pachyman, Tremella polysaccharide, Flammulina polysaccharide, Auricularia polysaccharide, Cordyceps polysaccharide, Phellopilus polysaccharide, and Coriolus polysaccharide.

[0029] Preferably, the animal polysaccharide includes any one or more of glycogen, chondroitin sulfate, hyaluronic acid, heparin, chitosan, chitin, or glycosaminoglycan. Preferably, the plant polysaccharide includes any one or more of cellulose, hemicellulose, pectin, inulin, konjac polysaccharide, astragalus polysaccharide, medlar polysaccharide, tea polysaccharide, kelp polysaccharide, or red date polysaccharide. Preferably, the algal polysaccharide includes any one or more of green algal polysaccharide, red algal polysaccharide, or brown algal polysaccharide.

[0030] Preferably, the mineral element includes, but is not limited to, any one or more of zinc, copper, iron, magnesium, calcium, or silicon.

[0031] In another aspect, the present application provides use of the cosmetic composition of any one of the above and the composition preparation of any one of the above in the preparation of a product selected from any one of the following products: (1) a cosmetic product for anti-aging, moisturizing, skin-whitening, or anti-oxidation; or (2) a health product for skin-whitening, anti-oxidation, or improving skin moisture; or (3) a pharmaceutical product for anti-aging, skin-whitening, improving skin moisture, anti-inflammation, or anti-oxidation.

[0032] In another aspect, the present application provides a cosmetic product comprising the cosmetic composition of any one of the above and the composition preparation of any one of the above.

[0033] In particular, the cosmetic product includes, but is not limited to, any one or more of face cream, face mask, eye cream, neck cream, hand cream, body lotion, moisturizing lotion, essence, toner, shampoo, hair conditioner, shower gel, or facial cleanser.

[0034] In particular, the cosmetic product further comprises a cosmetically acceptable auxiliary.

[0035] Preferably, the cosmetically acceptable auxiliary includes, but is not limited to, one or more of humectants, thickening agents, emollients, skin conditioning agents, emulsifiers, preservatives, emollients, lubricants, and stabilizers.

[0036] In another aspect, the present application provides a health product comprising the cosmetic composition or the composition preparation according to any one of the above.

[0037] In particular, the dosage form of the health product comprises a solid dosage form, a semi-solid dosage form or a liquid dosage form.

[0038] In particular, the health product further comprises a nutritionally acceptable nutritional additive.

[0039] Preferably, the nutritional additive comprises, but is not limited to, any one or more of dietary fiber, prebiotic, protein, lipid substance, mineral, vitamin.

[0040] In another aspect, the present application provides a pharmaceutical product comprising the cosmetic composition or the composition preparation according to any one of the above.

[0041] In particular, the pharmaceutical product further comprises a pharmaceutically acceptable excipient.

[0042] Preferably, the pharmaceutically acceptable excipient comprises, but is not limited to, a solvent, a diluent, a disintegrant, a precipitation inhibitor, a surfactant, a glidant, a binder, a lubricant, a dispersant, a suspending agent, an isotonic agent, a thickening agent, an emulsifying agent, a preservative, a stabilizer, a hydrating agent, an emulsification accelerator, a buffer, an absorbent, a coloring agent, a flavoring agent, a sweetening agent, an ion exchanger, a release agent, a coating agent, a flavoring agent or an antioxidant.

[0043] In particular, the dosage form of the pharmaceutical product comprises, but is not limited to, a gastrointestinal administration dosage form and a non-gastrointestinal administration dosage form according to the administration mode.

[0044] Preferably, the gastrointestinal administration dosage form comprises, but is not limited to, a tablet, a powder, a granule, a solution, a capsule, an emulsion, a suspension, an oil.

[0045] Preferably, the non-gastrointestinal administration dosage form comprises, but is not limited to, an injection administration dosage form, a respiratory administration dosage form, a skin administration dosage form, a mucosa administration dosage form and a cavity administration dosage form.

[0046] A method for anti-aging, moisturizing, whitening, moisturizing, improving skin moisture status, anti-inflammatory or antioxidant, the method comprising using the cosmetic composition, the composition preparation, the cosmetic product, the health product or the pharmaceutical product according to any one of the above.

[0047] The beneficial effects of the present application are: (1) The whitening composition of the present application can significantly increase the contents of collagen I, III, IV, VII and XVII; at the same time, inhibit collagen degradation enzyme MMP-1 and up-regulate the expression level of the pro-synthesis factor TGF-β1, which indicates that the whitening composition of the present application can multi-effect promote collagen regeneration.

[0048] (2) The whitening composition of the present application can significantly improve the contents of AQP3 and HA, strengthen the water-locking function of the skin, and has high efficient moisturizing effect.

[0049] (3) The whitening composition of the present application can activate the Nrf2 pathway, improve the expression level of SOD2, and inhibit the inflammatory factors IL-6 and IL-1a, and has strong antioxidant and anti-inflammatory effect. (4) The whitening composition of the present application combines collagen peptide, sodium hyaluronate and ergothioneine, and the prepared composition can have synergistic effect. BRIEF DESCRIPTION OF DRAWINGS

[0050] Figure 1 The figure is the thickness determination result of epidermal living cell layer; ## in the figure represents P<0.01 compared with the BC group; ** represents P<0.01 compared with the NC group.

[0051] Figure 2 The figure is the fibroblast density determination result of dermal layer; ## in the figure represents P<0.01 compared with the BC group; ** represents P<0.01 compared with the NC group.

[0052] Figure 3 The figure is the Collagen I expression determination result; ## in the figure represents P<0.01 compared with the BC group; ** represents P<0.01 compared with the NC group.

[0053] Figure 4 The figure is the Collagen III expression determination result; ## in the figure represents P<0.01 compared with the BC group; ** represents P<0.01 compared with the NC group.

[0054] Figure 5 The figure is the Collagen IV expression determination result; ## in the figure represents P<0.01 compared with the BC group; ** represents P<0.01 compared with the NC group.

[0055] Figure 6 The figure is the Collagen VII expression determination result; ## in the figure represents P<0.01 compared with the BC group; ** represents P<0.01 compared with the NC group.

[0056] Figure 7 The figure is the Collagen XVII expression determination result; ## in the figure represents P<0.01 compared with the BC group; ** represents P<0.01 compared with the NC group.

[0057] Figure 8 The figure is the collagen fiber content determination result; ## in the figure represents P<0.01 compared with the BC group; ** represents P<0.01 compared with the NC group.

[0058] Figure 9 The content of TGF-β1 is measured; ## in the figure represents P<0.01 compared with the BC group; ** represents P<0.01 compared with the NC group.

[0059] Figure 10 The content of MMP-1 is measured; ## in the figure represents P<0.01 compared with the BC group; ** represents P<0.01 compared with the NC group.

[0060] Figure 11 The content of HA is measured; ## in the figure represents P<0.01 compared with the BC group; ** represents P<0.01 compared with the NC group.

[0061] Figure 12 The content of Nrf2 is measured; ## in the figure represents P<0.01 compared with the BC group; ** represents P<0.01 compared with the NC group.

[0062] Figure 13 The content of SOD2 is measured; ## in the figure represents P<0.01 compared with the BC group; ** represents P<0.01 compared with the NC group.

[0063] Figure 14 The content of AQP3 is measured; ## in the figure represents P<0.01 compared with the BC group; ** represents P<0.01 compared with the NC group.

[0064] Figure 15 The content of IL-6 is measured; ## in the figure represents P<0.01 compared with the BC group; ** represents P<0.01 compared with the NC group.

[0065] Figure 16 The content of IL-1α is measured; ## in the figure represents P<0.01 compared with the BC group; ** represents P<0.01 compared with the NC group. DETAILED DESCRIPTION

[0066] The technical solutions in the embodiments of the present application will be described clearly and completely below in combination with the drawings in the embodiments of the present application. Obviously, the described embodiments are only some of the embodiments of the present application, rather than all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative work are within the scope of protection of the present application.

[0067] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. The terminology used herein in the description of the invention is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. The term "and / or" as used herein includes any and all combinations of one or more of the associated listed items.

[0068] Unless otherwise specified, the experimental methods used in the following examples are all routine experiments, and the materials and reagents used in the following examples are all commercially available unless otherwise specified.

[0069] Collagen peptides were purchased from Shanghai Tongyuan Food Technology Co., Ltd., batch number TC240621A2; Sodium hyaluronate was purchased from Shandong Huaxi Haiyu Biomedical Co., Ltd., batch number J202240289; Ergothioneine was purchased from Shanghai Ergothioneine Biotechnology Group Co., Ltd., batch number YGAF24022601.

[0070] Example 1: A whitening composition and its preparation method The composition consists of collagen peptides, sodium hyaluronate, and ergothioneine, with a mass concentration ratio of 80.647:4.839:2.016.

[0071] The preparation method is as follows: collagen peptides, sodium hyaluronate and ergothioneine are mixed in proportion and dissolved in water to obtain the product.

[0072] Example 2: A whitening composition and its preparation method The composition comprises collagen peptides, sodium hyaluronate, and ergothioneine, with a mass concentration ratio of 80.647:4.839:0.4032.

[0073] The preparation method is as follows: collagen peptides, sodium hyaluronate and ergothioneine are mixed in proportion and dissolved in water to obtain the product.

[0074] Example 3: A whitening composition and its preparation method The composition consists of collagen peptides, sodium hyaluronate, and ergothioneine, with a mass concentration ratio of 78.452:8.065:1.528.

[0075] The preparation method is as follows: collagen peptides, sodium hyaluronate and ergothioneine are mixed in proportion and dissolved in water to obtain the product.

[0076] Example 4: A whitening composition and its preparation method The composition consists of collagen peptides, sodium hyaluronate, and ergothioneine, with a mass concentration ratio of 83.148:3.550:2.845.

[0077] The preparation method is as follows: collagen peptides, sodium hyaluronate and ergothioneine are mixed in proportion and dissolved in water to obtain the product.

[0078] Example 5: A whitening composition and its preparation method The composition consists of collagen peptides, sodium hyaluronate, and ergothioneine, with a mass concentration ratio of 75.694:5.052:3.285.

[0079] The preparation method is as follows: collagen peptides, sodium hyaluronate and ergothioneine are mixed in proportion and dissolved in water to obtain the product.

[0080] Comparative Example 1: A whitening composition and its preparation method The composition consists of collagen peptides and ergothioneine in a mass concentration ratio of 85.486:2.016.

[0081] The preparation method is as follows: collagen peptides and ergothioneine are mixed in a certain proportion and dissolved in water to obtain the product.

[0082] Comparative Example 2: A whitening composition and its preparation method The composition consists of sodium hyaluronate and ergothioneine in a mass concentration ratio of 85.486:2.016.

[0083] The preparation method is as follows: Sodium hyaluronate and ergothioneine are mixed in a certain proportion and dissolved in water to obtain the product.

[0084] Effect Experiment: In the following test results data, the formulas for calculating the improvement rate and inhibition rate are as follows: Improvement rate (%) = [(Sample group - Negative control group) / Negative control group] × 100% Inhibition rate (%) = [(Negative control group - Sample group) / Negative control group] × 100% GraphPad Prism was used for plotting, and results are expressed as Mean ± SD. t-tests were used for comparisons between groups. All statistical analyses were two-tailed. P < 0.05 was considered statistically significant, and P < 0.01 was considered highly statistically significant.

[0085] Experimental Example 1: The ex vivo skin model was placed in a 6-well plate culture mold, and 3.7 mL of ex vivo skin model culture medium (Guangdong Boxi Biotechnology Shaanxi Branch) was added to each well. The plates were incubated at 37℃ in a 5% CO2 incubator, with the medium changed daily. After 2 days of culture, the ex vivo skin models were randomly divided into the blank group (BC), negative control group (NC), positive control group (PC), THA group, J group, ERG group, Example 1-Example 5 groups, and Comparative Example 1-Comparative Example 2 groups. Each group underwent the following drug treatments: Blank control (BC) group: Replace with fresh culture medium; Negative control (NC) group: Replace with fresh culture medium; Positive control (PC) group: Replace with fresh culture medium containing 0.01% m / v VC and 0.0007% m / v VE; THA group: Replace with fresh culture medium containing 0.087502% m / v sodium hyaluronate solution; Group J: Replace with fresh culture medium containing 0.087502% m / v collagen peptides; ERG group: Replace with fresh culture medium containing 0.087502% m / v ergothioneine; Example 1 group: Replace the fresh culture medium containing the composition of Example 1, that is, give fresh culture medium containing 0.080647% m / v collagen peptide, 0.004839% m / v sodium hyaluronate and 0.002016% m / v ergothioneine; Example 2 group: Replace the fresh culture medium containing the composition of Example 2, that is, give fresh culture medium containing 0.080647% m / v collagen peptide, 0.004839% m / v sodium hyaluronate and 0.0004032% m / v ergothioneine; Example 3 group: Replace the fresh culture medium containing the composition of Example 3, that is, give fresh culture medium containing 0.078452% m / v collagen peptide, 0.008065% m / v sodium hyaluronate and 0.001528% m / v ergothioneine; Example 4 group: Replace the fresh culture medium containing the composition of Example 4, that is, give fresh culture medium containing 0.083148% m / v collagen peptide, 0.00355% m / v sodium hyaluronate and 0.002845% m / v ergothioneine; Example 5 group: Replace the fresh culture medium containing the composition of Example 5, that is, give fresh culture medium containing 0.075694% m / v collagen peptide, 0.005052% m / v sodium hyaluronate and 0.003285% m / v ergothioneine; Comparative Example 1: Replace the culture medium with fresh medium containing the composition of Comparative Example 1, i.e., give fresh medium containing 0.085486% w / v collagen peptide and 0.002016% w / v ergothioneine. Comparative Example 2: Replace the culture medium with fresh medium containing the composition of Comparative Example 2, i.e., give fresh medium containing 0.085486% w / v sodium hyaluronate and 0.002016% w / v ergothioneine.

[0086] After completing the above drug administration treatment, all groups except BC were irradiated (30 J / cm²). 2 UVA +50mJ / cm 2 (UVB). After irradiation, the ex vivo skin model was incubated at 37°C in 5% CO2 medium for 24 hours. This procedure was repeated four times. After the repeated procedures, each group was treated with the drug daily for three days.

[0087] 1. Anti-wrinkle index: Epidermal cell layer thickness measurement Skin models used for testing were fixed with 4% paraformaldehyde for 24 hours. After fixation, H&E staining was performed, and images were taken and observed under a microscope. The thickness of the epidermal cell layer was collected and analyzed.

[0088] The measurement results are as follows Figure 1 As shown, compared with the BC group, the epidermal viable cell layer thickness in the NC group was significantly decreased, indicating that the stimulation conditions in this experiment were effective. Compared with the NC group, the epidermal viable cell layer thickness in the PC group was significantly increased, indicating that the positive control detection was effective. Compared with the NC group, the THA group, J group, ERG group, Comparative Example 1 group, Comparative Example 2 group, and Example 1-Example 5 groups all significantly increased the epidermal viable cell layer thickness. Among them, the average epidermal layer thickness of Example 1-Example 5 groups were 70.39 μm, 68.78 μm, 69.58 μm, 70.95 μm, and 70.97 μm, respectively, with an increase rate of 66.06%-71.34% compared with the NC group.

[0089] 2. Anti-wrinkle index: Determination of dermal cell thickness Skin models used for testing were fixed with 4% paraformaldehyde for 24 hours. After fixation, H&E staining was performed, and images were taken and observed under a microscope. The thickness of the dermal live cell layer was collected and analyzed.

[0090] The measurement results are as follows Figure 2As shown, compared with the BC group, the dermal fibroblast density in the NC group was significantly decreased, indicating that the stimulation conditions in this experiment were effective. Compared with the NC group, the dermal fibroblast density in the PC group was significantly increased, indicating that the positive control detection was effective. Compared with the NC group, the THA group, J group, ERG group, Comparative Example 1, Comparative Example 2, and Examples 1-5 groups all significantly increased the dermal fibroblast density. Among them, the dermal fibroblast density in Examples 1-5 groups increased by 191.67%-225% compared with the NC group.

[0091] 3. Anti-wrinkle index: Collagen I expression level determination Skin models used for testing were fixed in 4% paraformaldehyde for 24 hours. Immunofluorescence detection (Collagen I antibody, CST) was then performed, and images were taken and analyzed under a microscope. The green fluorescent region represents Collagen I; the stronger the green fluorescence, the higher the Collagen I content.

[0092] The measurement results are as follows Figure 3 As shown, compared with the BC group, the Collagen I content in the NC group was significantly decreased, indicating that the stimulation conditions in this experiment were effective. Compared with the NC group, the Collagen I content in the PC group was significantly increased, indicating that the positive control detection was effective. Compared with the NC group, the THA group, J group, ERG group, Comparative Example 1 group, Comparative Example 2 group, and Example 1-Example 5 groups all significantly increased the Collagen I content. Among them, the Collagen I content increase rate in Example 1-Example 5 groups was more significant than that in the other groups, with an increase rate of 190.91%-224.24% compared with the NC group.

[0093] 4. Anti-wrinkle index: Collagen III expression level determination Skin models used for testing were fixed with 4% paraformaldehyde for 24 hours, followed by immunofluorescence detection (Collagen III, Wuhan Sanying). Images were taken and analyzed under a microscope. The green fluorescent region represents Collagen III; stronger green fluorescence indicates a higher Collagen III content.

[0094] The measurement results are as follows Figure 4 As shown, compared with the NC group, the THA group, J group, ERG group, Comparative Example 1 group, Comparative Example 2 group, and Example 1-Example 5 groups all significantly increased the Collagen III content. Among them, the Collagen III content of Example 1-Example 5 groups increased by 256.67%-316.7% compared with the NC group.

[0095] 5. Anti-wrinkle index: Collagen IV expression level determination Skin models used for testing were fixed in 4% paraformaldehyde for 24 hours. Immunofluorescence detection (Collagen IV antibody, Abcam) was then performed, and images were taken and analyzed under a microscope. The green fluorescent region represents Collagen IV; the stronger the green fluorescence, the higher the Collagen IV content.

[0096] The measurement results are as follows Figure 5 As shown, compared with the NC group, the THA group, J group, ERG group, Comparative Example 1 group, Comparative Example 2 group, and Example 1-Example 5 groups all significantly increased the Collagen IV content. Among them, the Collagen IV content increase rate of Example 1-Example 5 groups was more significant than that of the other groups, with an increase rate of 155.81%-202.33% compared with the NC group.

[0097] 6. Anti-wrinkle index: Collagen VII expression level determination Skin models used for testing were fixed in 4% paraformaldehyde for 24 hours, followed by immunofluorescence detection (Collagen VII, Abcam). Images were taken and analyzed under a microscope. The green fluorescent region represents Collagen VII; stronger green fluorescence indicates a higher Collagen VII content.

[0098] The measurement results are as follows Figure 6 As shown, compared with the NC group, the THA group, J group, ERG group, Comparative Example 1 group, Comparative Example 2 group and Example 1-Example 5 group all significantly increased the Collagen VII content. Among them, the Collagen VII content increase rate of Example 1-Example 5 group was more obvious than that of the other groups, with an increase rate of 125%-140.91% compared with the NC group.

[0099] 7. Anti-wrinkle index: Collagen XVII expression level determination Skin models used for testing were fixed in 4% paraformaldehyde for 24 hours. Immunofluorescence detection (using Collagen XVII antibody, Abcam) was then performed, and images were taken and analyzed under a microscope. The green fluorescent region represents Collagen XVII; stronger green fluorescence indicates a higher Collagen XVII content.

[0100] The measurement results are as follows Figure 7As shown, compared with the NC group, the THA group, J group, ERG group, Comparative Example 1 group, Comparative Example 2 group, and Example 1-Example 5 groups all significantly increased the content of Collagen XVII. Among them, the increase rate of Collagen XVII content in Example 1-Example 5 groups was more significant than that in the other groups, with an increase rate of 163.41%-190.2% compared with the NC group.

[0101] 8. Anti-wrinkle index: Collagen fiber content determination Skin models used for testing were fixed with 4% paraformaldehyde, stained with Masson's stain, and photographed under a microscope. The blue area represents collagen fibers; the larger the blue area, the higher the collagen fiber content.

[0102] The measurement results are as follows Figure 8 As shown, compared with the BC group, the collagen fiber content in the NC group was significantly lower, indicating that the stimulation conditions in this experiment were effective. Compared with the NC group, the collagen fiber content in the PC group was significantly higher, indicating that the positive control test was effective. Compared with the NC group, the THA group, J group, ERG group, Comparative Example 1 group, Comparative Example 2 group, and Example 1-Example 5 groups all significantly increased collagen fiber content. Among them, the collagen fiber increase rate of Example 1-Example 5 groups was more significant than that of the other groups, with an increase rate of 125.58%-148.84% compared with the NC group.

[0103] 9. Anti-wrinkle index: TGF-β1 content determination Skin models used for testing were fixed in 4% paraformaldehyde for 24 hours. Immunofluorescence detection (TGF-β1 antibody, Abcam) was then performed, and images were taken and analyzed under a microscope. The green fluorescent region represents TGF-β1; the stronger the green fluorescence, the higher the TGF-β1 content.

[0104] The measurement results are as follows Figure 9 As shown, compared with the BC group, the TGF-β1 content in the NC group was significantly decreased, indicating that the stimulation conditions in this experiment were effective. Compared with the NC group, the TGF-β1 content in the PC group was significantly increased, indicating that the positive control detection was effective. Compared with the NC group, the THA group, J group, ERG group, Comparative Example 1, Comparative Example 2, and Example 1-Example 5 groups all significantly increased TGF-β1 expression. Among them, the TGF-β1 expression increase rate in Example 1-Example 5 groups was more significant than that in the other groups, with an increase rate of 197.44%-225.64% compared with the NC group.

[0105] 10. Anti-wrinkle index: MMP-1 content determination After the administration of the drug, the skin model culture medium was collected in centrifuge tubes. After collection, the samples used for ELISA detection were frozen at -80°C and analyzed according to the instructions of the MMP-1 ELISA kit (Abcam).

[0106] The measurement results are as follows Figure 10 As shown, compared with the BC group, the MMP-1 content in the NC group was significantly increased, indicating that the stimulation conditions in this experiment were effective. Compared with the NC group, the MMP-1 content in the PC group was significantly decreased, indicating that the positive control detection was effective. Compared with the NC group, the THA group, J group, ERG group, Comparative Example 1, Comparative Example 2, and Example 1-Example 5 groups all significantly reduced MMP-1 expression. Among them, Example 1-Example 5 groups showed the most significant inhibition of MMP-1 expression, with inhibition rates of 44.89%-46.2% compared with the NC group.

[0107] 11. Moisturizing index: HA content determination Biotin coloration assay: A skin model for testing was taken and fixed with 4% paraformaldehyde for 24 hours. The images were then photographed and analyzed (Hyaluronan Binding Protein, Biotin Conj., Amsbio). Green fluorescence represents HA; the stronger the green fluorescence, the higher the HA content.

[0108] The measurement results are as follows Figure 11 As shown, compared with the BC group, the HA content in the NC group decreased significantly, indicating that the stimulation conditions in this experiment were effective. Compared with the NC group, the HA content in the PC group increased significantly, indicating that the positive control detection was effective. Compared with the NC group, the THA group, J group, ERG group, Comparative Example 1 group, Comparative Example 2 group, and Example 1-Example 5 groups all significantly increased the HA content. Among them, the improvement effect of Example 1-Example 5 groups was more obvious, with an improvement rate of 272.41%-293.10% compared with the NC group.

[0109] 12. Antioxidant index: Nrf2 content determination Skin models used for testing were fixed in 4% paraformaldehyde for 24 hours. Immunofluorescence detection (Nrf2 antibody, Abcam) was then performed, and images were taken and analyzed under a microscope. The green fluorescent region represents Nrf2; the stronger the green fluorescence, the higher the Nrf2 content.

[0110] The measurement results are as follows Figure 12As shown, compared with the BC group, the Nrf2 content in the NC group was significantly decreased, indicating that the stimulation conditions in this experiment were effective. Compared with the NC group, the Nrf2 content in the PC group was significantly increased, indicating that the positive control detection was effective. Compared with the NC group, the THA group, J group, ERG group, Comparative Example 1 group, Comparative Example 2 group, and Example 1-Example 5 groups all significantly increased the Nrf2 content. Among them, the improvement effect of Example 1-Example 5 groups was more obvious, with an improvement rate of 296.55%-317.24% compared with the NC group.

[0111] 13. Antioxidant index: SOD2 content determination Skin models used for testing were fixed in 4% paraformaldehyde for 24 hours. Immunofluorescence detection (SOD2 antibody, Abcam) was then performed, and images were taken and analyzed under a microscope. The green fluorescent region represents SOD2; the stronger the green fluorescence, the higher the SOD2 content.

[0112] The measurement results are as follows Figure 13 As shown, compared with the BC group, the SOD2 content in the NC group decreased significantly, indicating that the stimulation conditions in this experiment were effective. Compared with the NC group, the SOD2 content in the PC group increased significantly, indicating that the positive control detection was effective. Compared with the NC group, the THA group, J group, ERG group, Comparative Example 1 group, Comparative Example 2 group, and Example 1-Example 5 groups all significantly increased the SOD2 content. Among them, the improvement effect of Example 1-Example 5 groups was more obvious, with an improvement rate of 264.29%-307.14% compared with the NC group.

[0113] Experimental Example 2 The ex vivo skin model was placed in a 6-well plate culture mold, and 3.7 mL of ex vivo skin model culture medium (Guangdong Boxi Biotechnology Shaanxi Branch) was added to each well. The plates were incubated at 37℃ in a 5% CO2 incubator, with the medium changed daily. After 2 days of culture, the ex vivo skin models were randomly divided into the blank group (BC), negative control group (NC), positive control group (PC), THA group, J group, ERG group, Example 1-Example 5 groups, and Comparative Example 1-Comparative Example 2 groups. Each group underwent the following drug treatments: Blank control (BC) group: Replace with fresh culture medium; Negative control (NC) group: Replace with fresh culture medium; Positive control (PC) group: Replace with fresh culture medium containing 0.0033% m / v CaCl2; THA group: Replace with fresh culture medium containing 0.087502% m / v sodium hyaluronate solution; Group J: Replace with fresh culture medium containing 0.087502% m / v collagen peptides; ERG group: Replace with fresh culture medium containing 0.087502% m / v ergothioneine; Example 1 group: Replace the fresh culture medium containing the composition of Example 1, that is, give fresh culture medium containing 0.080647% m / v collagen peptide, 0.004839% m / v sodium hyaluronate and 0.002016% m / v ergothioneine; Example 2 group: Replace the fresh culture medium containing the composition of Example 2, that is, give fresh culture medium containing 0.080647% m / v collagen peptide, 0.004839% m / v sodium hyaluronate and 0.0004032% m / v ergothioneine; Example 3 group: Replace the fresh culture medium containing the composition of Example 3, that is, give fresh culture medium containing 0.078452% m / v collagen peptide, 0.008065% m / v sodium hyaluronate and 0.001528% m / v ergothioneine; Example 4 group: Replace the fresh culture medium containing the composition of Example 4, that is, give fresh culture medium containing 0.083148% m / v collagen peptide, 0.00355% m / v sodium hyaluronate and 0.002845% m / v ergothioneine; Example 5 group: Replace the fresh culture medium containing the composition of Example 5, that is, give fresh culture medium containing 0.075694% m / v collagen peptide, 0.005052% m / v sodium hyaluronate and 0.003285% m / v ergothioneine; Comparative Example 1: Replace the culture medium with fresh medium containing the composition of Comparative Example 1, i.e., give fresh medium containing 0.085486% w / v collagen peptide and 0.002016% w / v ergothioneine. Comparative Example 2: Replace the culture medium with fresh medium containing the composition of Comparative Example 2, i.e., give fresh medium containing 0.085486% w / v sodium hyaluronate and 0.002016% w / v ergothioneine.

[0114] After completing the above drug administration treatment, all groups except BC were irradiated (30 J / cm²). 2 UVA +50mJ / cm 2 (UVB). After irradiation, the ex vivo skin model was incubated at 37°C in 5% CO2 medium for 24 hours. This procedure was repeated four times. After the repeated procedures, each group was treated with the drug daily for three days.

[0115] Moisturizing index: AQP3 content determination Skin models used for testing were fixed in 4% paraformaldehyde for 24 hours. Immunofluorescence detection (AQP3 antibody, Abcam) was then performed, and images were taken and analyzed under a microscope. The green fluorescent region represents AQP3; the stronger the green fluorescence, the higher the AQP3 content.

[0116] The measurement results are as follows Figure 14 As shown, compared with the BC group, the AQP3 content in the NC group was significantly lower, indicating that the stimulation conditions in this experiment were effective. Compared with the NC group, the AQP3 content in the PC group was significantly higher, indicating that the positive control detection was effective. Compared with the NC group, the THA group, J group, ERG group, Comparative Example 1 group, Comparative Example 2 group, and Example 1-Example 5 groups all significantly increased the AQP3 content. Among them, the improvement effect of Example 1-Example 5 groups was more obvious, with an improvement rate of 138.1%-180.95% compared with the NC group.

[0117] Experimental Example 3 The ex vivo skin model was placed in a 6-well plate culture mold, and 3.7 mL of ex vivo skin model culture medium (Guangdong Boxi Biotechnology Shaanxi Branch) was added to each well. The plates were incubated at 37℃ in a 5% CO2 incubator, with the medium changed daily. After 2 days of culture, the ex vivo skin models were randomly divided into the blank group (BC), negative control group (NC), positive control group (PC), THA group, J group, ERG group, Example 1-Example 5 groups, and Comparative Example 1-Comparative Example 2 groups. Each group underwent the following drug treatments: Blank control (BC) group: Replace with fresh culture medium; Negative control (NC) group: Replace with fresh culture medium; Positive control (PC) group: Replace with fresh culture medium containing 0.01% m / v dexamethasone; THA group: Replace with fresh culture medium containing 0.087502% m / v sodium hyaluronate solution; Group J: Replace with fresh culture medium containing 0.087502% m / v collagen peptides; ERG group: Replace with fresh culture medium containing 0.087502% m / v ergothioneine; Example 1 group: Replace the fresh culture medium containing the composition of Example 1, that is, give fresh culture medium containing 0.080647% m / v collagen peptide, 0.004839% m / v sodium hyaluronate and 0.002016% m / v ergothioneine; Example 2 group: Replace the fresh culture medium containing the composition of Example 2, that is, give fresh culture medium containing 0.080647% m / v collagen peptide, 0.004839% m / v sodium hyaluronate and 0.0004032% m / v ergothioneine; Example 3 group: Replace the fresh culture medium containing the composition of Example 3, that is, give fresh culture medium containing 0.078452% m / v collagen peptide, 0.008065% m / v sodium hyaluronate and 0.001528% m / v ergothioneine; Example 4 group: Replace the fresh culture medium containing the composition of Example 4, that is, give fresh culture medium containing 0.083148% m / v collagen peptide, 0.00355% m / v sodium hyaluronate and 0.002845% m / v ergothioneine; Example 5 group: Replace the fresh culture medium containing the composition of Example 5, that is, give fresh culture medium containing 0.075694% m / v collagen peptide, 0.005052% m / v sodium hyaluronate and 0.003285% m / v ergothioneine; Comparative Example 1: Replace the culture medium with fresh medium containing the composition of Comparative Example 1, i.e., give fresh medium containing 0.085486% w / v collagen peptide and 0.002016% w / v ergothioneine. Comparative Example 2: Replace the culture medium with fresh medium containing the composition of Comparative Example 2, i.e., give fresh medium containing 0.085486% w / v sodium hyaluronate and 0.002016% w / v ergothioneine.

[0118] After completing the above drug administration treatment, all groups except BC were irradiated (30 J / cm²). 2 UVA +50mJ / cm 2 (UVB). After irradiation, the ex vivo skin model was incubated at 37°C in 5% CO2 medium for 24 hours. This procedure was repeated four times. After the repeated procedures, each group was treated with the drug daily for three days.

[0119] After drug administration, the skin model culture medium was collected in centrifuge tubes. After collection, the samples used for ELISA detection were frozen at -80°C and analyzed according to the instructions of the IL-1α ELISA kit (Abcam) and IL-6 ELISA kit (Abcam).

[0120] 1. Anti-inflammatory marker: IL-6 content measurement results The measurement results are as follows Figure 15As shown, compared with the BC group, the IL-6 level in the NC group was significantly increased, indicating that the stimulation conditions in this experiment were effective. Compared with the NC group, the IL-6 level in the PC group was significantly decreased, indicating that the positive control detection was effective. Compared with the NC group, the THA group, J group, ERG group, Comparative Example 1 group, Comparative Example 2 group, and Example 1-Example 5 groups all significantly reduced IL-6 expression levels, with the improvement effect of Example 1-Example 5 groups being more significant than the other groups, with inhibition rates of 60.19%-62.90% compared with the NC group.

[0121] 2. Anti-inflammatory marker: IL-1α content determination The measurement results are as follows Figure 16 As shown, compared with the BC group, the IL-1α level in the NC group was significantly increased, indicating that the stimulation conditions in this experiment were effective. Compared with the NC group, the IL-1α level in the PC group was significantly decreased, indicating that the positive control detection was effective. Compared with the NC group, the THA group, J group, ERG group, Comparative Example 1, Comparative Example 2, and Example 1-Example 5 groups all significantly reduced IL-1α expression, with the improvement effect of Example 1-Example 5 groups being more significant than the other groups, with inhibition rates of 59.85%-62.8% compared with the NC group.

[0122] The above detailed description is a specific illustration of one feasible embodiment of the present invention, and this embodiment is not intended to limit the patent scope of the present invention. It should be noted that all equivalent implementations or modifications made without departing from the present invention should be included within the scope of the technical solution of the present invention. Therefore, the protection scope of the present invention should be determined by the appended claims.

Claims

1. A cosmetic composition, characterized in that: The cosmetic composition comprises collagen peptides, sodium hyaluronate, and ergothioneine, wherein the mass concentration ratio of collagen peptides, sodium hyaluronate, and ergothioneine is 75-85: 3.5-10：0.4-3.5。 2. The cosmetic composition according to claim 1, characterized in that: The mass concentration ratio of the collagen peptides, sodium hyaluronate, and ergothioneine is 80-81: 4.7-4.9：2.0-2.3。 3. The cosmetic composition according to claim 1, characterized in that: The mass concentration ratio of the collagen peptide, sodium hyaluronate, and ergothioneine is 80.647:4.839:2.

016.

4. The cosmetic composition according to claim 1, characterized in that: The collagen peptides mentioned are collagen peptides containing 20%-100% collagen tripeptides; the ergothioneine mentioned is L-ergothioneine.

5. The cosmetic composition according to claim 1, characterized in that, The cosmetic composition further includes a solvent, which is selected from one or more of the following: water, polyol solvents, alcohol solvents, and oil solvents.

6. A method for preparing the cosmetic composition according to any one of claims 1-5, characterized in that, The preparation method includes mixing collagen peptides, sodium hyaluronate and ergothioneine in a certain proportion; or mixing collagen peptides, sodium hyaluronate and ergothioneine in a certain proportion and dissolving them in a solvent.

7. A compositional formulation comprising the cosmetic composition according to any one of claims 1-5.

8. The composition formulation according to claim 7, characterized in that, The composition formulation further includes any one or more of the following: plant extracts, vitamins, bioactive peptides, enzymes, acids, polysaccharides, or mineral elements.

9. The use of the cosmetic composition according to any one of claims 1-5 or the composition formulation according to any one of claims 7-8 in the preparation of a product, characterized in that, The product mentioned is selected from any one of the following products: (1) Cosmetics that are anti-aging, hydrating, moisturizing, anti-inflammatory, whitening, or antioxidant; Or (2) anti-inflammatory, whitening, antioxidant, or skin hydration-enhancing health products; Or (3) Anti-aging, whitening, skin moisture improvement, anti-inflammatory or antioxidant drugs.

10. A cosmetic product, characterized in that, The cosmetic comprises the composition according to any one of claims 1-5 or the composition formulation according to any one of claims 7-8.

11. The cosmetic product according to claim 10, characterized in that, The cosmetics mentioned include any one or more of the following: face cream, face mask, eye cream, neck cream, hand cream, body lotion, moisturizer, serum, toner, shampoo, conditioner, shower gel, or facial cleanser.

12. The cosmetic product according to claim 10, characterized in that, The cosmetics mentioned also include excipients acceptable in the cosmetics industry.

13. The cosmetic product according to claim 12, characterized in that, The cosmetic excipients acceptable in the field include one or more of the following: moisturizers, thickeners, emollients, skin conditioning agents, emulsifiers, preservatives, softeners, lubricants, and stabilizers.

14. A health product, characterized in that, The cosmetic comprises the composition according to any one of claims 1-5 or the composition formulation according to any one of claims 7-8.

15. The health product according to claim 14, characterized in that, The dosage forms of the health products include solid dosage forms, semi-solid dosage forms, or liquid dosage forms.

16. The health product according to claim 14, characterized in that, The health products mentioned also include nutritionally acceptable nutritional additives.

17. The health product according to claim 16, characterized in that, The nutritional additives include any one or more of the following: dietary fiber, prebiotics, protein, lipids, minerals, and vitamins.

18. A medicine, characterized in that, The pharmaceutical product comprises the composition according to any one of claims 1-5 or the composition formulation according to any one of claims 7-8.

19. The pharmaceutical product according to claim 18, characterized in that, The medicine also includes pharmaceutically acceptable excipients.

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