Skin care composition and application thereof

Abstract

The invention discloses a skin care composition and application thereof.The skin care composition contains acetylchitosamine and succinic acid, acetylchitosamine and succinic acid are compounded, succinic acid serves as mild cutin-removing organic acid, cutin is softened by helping to dissolve tight connection between keratinocytes, and the skin care effect is improved. The composition helps to open channels for active ingredients to permeate into the skin, improves skin permeation of the water-soluble active ingredient acetylchitosamine, and meanwhile, due to intermolecular interaction of acetylchitosamine and succinic acid, the composition can show a synergistic effect on whitening and softening cutin.

Description

Technical Field

[0001] The invention relates to a skin care composition and application thereof, belonging to the technical field of cosmetics. Background Art

[0002] The skin is one of the largest organs in the human body. It covers the outside of the body and plays an important role in protecting internal tissues and organs from external damage. When the skin is exposed to ultraviolet rays, the pigment cells in the skin, melanocytes, produce melanin, which can absorb and scatter ultraviolet rays, thereby preventing them from damaging skin tissue. At the same time, the deposition of melanin also causes skin problems such as dull skin and spots, affecting the appearance.

[0003] In order to improve skin brightness and remove melanin, removing the stratum corneum is an effective way. As an active ingredient in the field of skin care, N-Acetylglucosamine (NAG) has gradually attracted attention from the scientific community for its effects on softening the stratum corneum and regulating sebum secretion. As a monomer that makes up hyaluronic acid, NAG participates in the synthesis of hyaluronic acid, enhances the water retention of the epidermis, and thus improves the softness of the stratum corneum and the overall skin quality.

[0004] Succinic acid is an intermediate in the tricarboxylic acid cycle and has been found to have good effects in skin care research in recent years. Succinic acid can inhibit tyrosinase activity and reduce the synthesis of melanin by affecting intracellular signal transduction, thereby fighting pigmentation and achieving a whitening effect. At the same time, succinic acid, as a mild exfoliant, can promote keratinocyte renewal, improve skin quality, and increase skin smoothness. Summary of the invention

[0005] The object of the present invention is to provide a skin care composition, which contains acetyl glucosamine and succinic acid. Experimental verification shows that the combination of the two exhibits a synergistic effect in whitening and softening keratin.

[0006] The present invention provides a skin care composition, and its technical solution is as follows:

[0007] A skin care composition comprises acetyl glucosamine and succinic acid in a physiologically acceptable medium.

[0008] Furthermore, the mass ratio of acetyl glucosamine to succinic acid is 0.01-50:1.

[0009] Furthermore, the mass ratio of acetyl glucosamine to succinic acid is 0.1-50:1.

[0010] Furthermore, the mass ratio of acetyl glucosamine to succinic acid is 0.5-50:1.

[0011] Furthermore, the mass ratio of acetyl glucosamine to succinic acid is 1-45:1.

[0012] Furthermore, the mass ratio of acetyl glucosamine to succinic acid is 2-45:1.

[0013] Furthermore, the mass ratio of acetyl glucosamine to succinic acid is 2.5-45:1.

[0014] Furthermore, the mass ratio of acetyl glucosamine to succinic acid is 3-45:1.

[0015] Furthermore, the mass percentage of acetyl glucosamine in the skin care composition is 0.001%-10%.

[0016] Furthermore, the mass percentage of succinic acid in the skin care composition is 0.002%-5%.

[0017] Furthermore, the skin care composition also contains other acceptable active ingredients and / or auxiliary materials.

[0018] The present invention also provides application of the skin care composition in preparing cosmetics and foods.

[0019] The present invention also provides the use of a skin care composition containing acetyl glucosamine and succinic acid in the synergistic preparation of a product for inhibiting tyrosinase activity and / or inhibiting melanin synthesis.

[0020] Furthermore, the mass ratio of the acetyl glucosamine to succinic acid is 0.5-50:1.

[0021] The invention also provides application of a skin care composition containing acetyl glucosamine and succinic acid in synergistic skin whitening.

[0022] Furthermore, the mass ratio of the acetyl glucosamine to succinic acid is 0.5-50:1.

[0023] The present invention also provides the use of a skin care composition containing acetyl glucosamine and succinic acid in the synergistic preparation of a product for reducing corneal desmosin (CDSN).

[0024] Furthermore, the mass ratio of the acetyl glucosamine to succinic acid is 0.5-50:1.

[0025] The present invention also provides the use of a skin care composition containing acetyl glucosamine and succinic acid in synergistically softening skin keratin and / or exfoliating.

[0026] Furthermore, the mass ratio of the acetyl glucosamine to succinic acid is 0.5-50:1.

[0027] The present invention has the following beneficial effects:

[0028] The present invention compounds acetyl glucosamine with succinic acid to exhibit synergistic effects in whitening and / or softening keratin and / or exfoliating. DETAILED DESCRIPTION

[0029] The following is a description of exemplary embodiments of the present invention, including various details of the embodiments of the present invention to aid understanding, which should be considered as merely exemplary. Therefore, it should be appreciated by those of ordinary skill in the art that various changes and modifications may be made to the embodiments described herein without departing from the scope and spirit of the present invention. Likewise, for clarity and simplicity, descriptions of well-known functions and structures are omitted in the following description.

[0030] The words "include" or "comprising" mentioned throughout the specification and claims are open-ended terms and should be interpreted as "including but not limited to". The following description of the specification is a preferred embodiment of the present application, but the description is for the purpose of the general principles of the specification and is not intended to limit the scope of the present application. The scope of protection of the present application shall be determined by the attached claims.

[0031] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those of ordinary skill in the art to which the present disclosure belongs. Although methods and materials similar or identical to those described herein can be used in experiments or practical applications, the present invention still describes the materials and methods below. In the event of conflict, the present specification, including definitions, will prevail.

[0032] The present invention relates to a composition, and the "composition" can achieve the effects of the present invention, and the "composition" includes but is not limited to using the components simultaneously or sequentially. The "simultaneous use" includes using them together in the same preparation or using them separately in different preparations. The "sequential use" includes using them sequentially in different preparations, and there is no restriction on the order of sequential use.

[0033] As used herein, the term "and / or" in phrases such as "A and / or B" is intended to include both A and B; A or B; A (alone); and B (alone). Similarly, as used herein, the term "and / or" in phrases such as "A, B, and / or C" is intended to include each of the following embodiments: A, B, and C; A, B, or C; A or C; A or B; B or C; A and C; A and B; B and C; A (alone); B (alone); and C (alone).

[0034] The present invention provides a skin care composition, which comprises a physiologically acceptable medium, wherein acetyl glucosamine and succinic acid are contained in the physiologically acceptable medium. Acetyl glucosamine and succinic acid are compounded to synergistically inhibit tyrosinase activity and reduce the synthesis of melanin, reduce the content of corneal desmosin connecting cells, and show a synergistic effect in whitening and softening keratin.

[0035] In the experiment, it was found that when the mass ratio of acetyl glucosamine to succinic acid was 0.01-50:1, it showed obvious synergistic effects in skin whitening and softening keratin, such as 0.01:1, 0.02:1, 0.03:1, 0.04:1, 0.05:1, 0.06:1, 0.07:1, 0.08:1, 0.09:1, 0.1:1, 0.2:1, 0.3:1, 0.4:1, 0.5:1, 0.6:1, 0.7:1, 0.8:1, 0.9:1, 1:1, 2:1, 2.5:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1 :1, 10:1, 11:1, 12:1, 13:1, 14:1, 15:1, 16:1, 17:1, 18:1, 19:1, 20:1, 21:1, 22:1, 23:1, 24:1, 25:1, 26:1, 27:1, 28:1, 29:1, 30:1, 31:1, 32:1, 33:1, 34:1, 35:1, 36:1, 37:1, 38:1, 39:1, 40:1, 41:1, 42:1, 43:1, 44:1, 45:1, 46:1, 47:1, 48:1, 49:1, 50:1 and any range therebetween.

[0036] Furthermore, the mass ratio of acetyl glucosamine to succinic acid is 0.1-50:1.

[0037] Furthermore, the mass ratio of acetyl glucosamine to succinic acid is 0.5-50:1.

[0038] Furthermore, the mass ratio of acetyl glucosamine to succinic acid is 0.5-45:1.

[0039] Furthermore, the mass ratio of acetyl glucosamine to succinic acid is 1-45:1.

[0040] Furthermore, the mass ratio of acetyl glucosamine to succinic acid is 2-45:1.

[0041] Furthermore, the mass ratio of acetyl glucosamine to succinic acid is 2.5-45:1.

[0042] Furthermore, the mass ratio of acetyl glucosamine to succinic acid is 3-45:1.

[0043] In a specific embodiment of the present invention, the mass percentage of acetyl glucosamine in the skin care composition is 0.001%-10%, for example, 0.001%, 0.0015%, 0.002%, 0.0025%, 0.003%, 0.0035%, 0.004%, 0.0045%, 0.005%, 0.0055%, 0.006%, 0.0065%, 0.007%, 0.0075%, 0.008%, 0.0085%, 0.009%, 0.0095%, 0.01%, 0.0 2%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1.0%, 1.5%, 2.0%, 2.5%, 3.0%, 3.5%, 4.0%, 4.5%, 5.0%, 5.5%, 6.0%, 6.5%, 7.0%, 7.5%, 8.0%, 8.5%, 9.0%, 9.5%, 10% and any range in between.

[0044] In a specific embodiment of the present invention, the mass percentage of succinic acid in the skin care composition is 0.002%-5%, for example, 0.002%, 0.0025%, 0.003%, 0.0035%, 0.004%, 0.0045%, 0.005%, 0.0055%, 0.006%, 0.0065%, 0.007%, 0.0075%, 0.008%, 0.0085%, 0.009%, 0.010%, 0.011%, 0.012%, 0.013%, 0.014%, 0.015%, 0.016%, 0.017%, 0.018%, 0.019%, 0.199%, 10.25%, 10.35%, 10.45%, 10.5 0.095%, 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1.0%, 1.5%, 2.0%, 2.5%, 3.0%, 3.5%, 4.0%, 4.5%, 5.0% and any range in between.

[0045] The acetyl glucosamine of the present invention is also known as N-acetyl-D-glucosamine, N-acetylglucosamine, or NAG for short, and is an amino monosaccharide generated by replacing the -OH group at the 2-position of the glucose molecule with an acetylamino group. Acetyl glucosamine exists naturally in human bones, articular cartilage, skin and other tissues, and can also be obtained from the outside. Acetyl glucosamine is commonly found in crustaceans, such as shrimp, crab, oyster, etc. The acetyl glucosamine can be introduced into food and skin care compositions in any form, such as a pure product, a solution state, or an extract state containing acetyl glucosamine.

[0046] The succinic acid of the present invention is also known as succinic acid, is a dicarboxylic acid, is colorless crystal, and tastes sour. The succinic acid can be introduced into food and skin care compositions in any form, such as pure product, solution state, or extract state containing succinic acid.

[0047] In a specific embodiment of the present invention, the skin care composition consists of acetyl glucosamine and succinic acid.

[0048] In a specific embodiment of the present invention, the skin care composition includes, in addition to acetyl glucosamine and succinic acid, other active ingredients and / or adjuvants acceptable in the field of food or cosmetics. The other active ingredients may be active ingredients having at least one effect of whitening, moisturizing, anti-acne, anti-inflammatory, antibacterial, anti-aging, anti-oxidation, anti-wrinkle, etc. The cosmetic adjuvant may be an adjuvant of at least one of a moisturizer, an emulsifier, a chelating agent, a preservative, a thickener, a grease, a solvent, a surfactant, an antioxidant, a pH regulator, a functional additive, a fragrance, a pigment, etc. The food adjuvant may be an adjuvant of at least one of a preservative, a sweetener, an antioxidant, a flavor enhancer, an emulsifier, a thickener, a leavening agent, and a colorant.

[0049] In a specific embodiment of the present invention, the skin care composition is a cosmetic and can be made into different types, including but not limited to essence water, lotion, cream, sunscreen, hand mask, hand cream, shower gel, shampoo, etc.

[0050] In a specific embodiment of the present invention, the skin care composition is a food, which can be made into different types, including but not limited to candies, biscuits, cakes, beverages, convenience foods, etc.

[0051] The present invention also provides a method for preparing the above skin care composition, which comprises mixing the components uniformly. The mixing method and order of the components are not limited.

[0052] The present invention also provides the use of the skin care composition in the synergistic preparation of products for inhibiting tyrosinase activity and / or inhibiting melanin synthesis.

[0053] The present invention also provides the use of the skin care composition in the synergistic preparation of a product for reducing corneal desmosin (CDSN).

[0054] The present invention also provides the use of the skin care composition in synergistic skin whitening, and the application mode can be used as a raw material in the preparation of skin whitening products, or used in skin whitening in combination with corresponding beauty equipment or massage techniques.

[0055] The present invention also provides the use of the above-mentioned skin care composition in synergistically softening skin keratin and / or exfoliating. The application method can be used as a raw material in the preparation of products for softening skin keratin and / or exfoliating, or used in conjunction with corresponding beauty equipment or massage techniques for softening skin keratin and / or exfoliating.

[0056] The present invention also provides the use of the skin care composition in the preparation of cosmetics, and the application mode thereof can be used as a raw material in the preparation of cosmetics.

[0057] In a specific embodiment of the present invention, the cosmetic is a cosmetic having skin whitening effect.

[0058] In a specific embodiment of the present invention, the cosmetic is a cosmetic having the effect of softening skin keratin and / or exfoliating.

[0059] The present invention also provides a cosmetic comprising the skin care composition.

[0060] The present invention also provides the use of the skin care composition in preparing food, and the use mode thereof can be used as a raw material in preparing food.

[0061] In a specific embodiment of the present invention, the food is a food with skin whitening effect.

[0062] The present invention also provides a food, which comprises the skin care composition.

[0063] The present invention produces a synergistic effect in whitening and / or softening keratin and / or exfoliating after compounding acetyl glucosamine with succinic acid.

[0064] Example

[0065] The efficacy of the compound of acetyl glucosamine and succinic acid was verified below. In the following experiments, unless otherwise specified, the concentrations are all mass percentage concentrations, and the reagents or instruments without indicating the manufacturer are all conventional reagent products that can be purchased commercially.

[0066] In the following experiments, the reagents and instruments used are as follows:

[0067] name factory Remark Acetyl glucosamine Bloomage Biopharmaceuticals Purity>99% Succinic acid Roquette, France Purity>99% Deionized water Laboratory-made PBS Vivacell DMSO Sinopharm NP-40 Lysate Blue Sky Kojic Acid Sinopharm Purity>99% Levodopa Sinopharm Paraformaldehyde Seville Dewaxing liquid Seville Primary antibody (goat source) ABCAM Secondary antibody (goat source) ABCAM Autofluorescence Quencher Solution B Seville Anti-fluorescence fading mounting medium Seville BSA bovine serum albumin Merck DAPI stain Blue Sky <![CDATA[CO 2 Incubator]]> Thermo, 160i Biological Safety Cabinets Su Jingantai, BSC-1604II A2

[0068] Example 1 Test on the efficacy of NAG, succinic acid and their combination in inhibiting tyrosinase activity and melanin synthesis

[0069] This test uses a melanocyte model to measure the effects of different samples on the melanin synthesis inhibition rate and tyrosinase synthesis inhibition rate.

[0070] 1. Experimental samples

[0071] The contents of effective ingredients in the experimental samples are shown in Table 1 below.

[0072] Table 1 Composition and concentration of each experimental sample

[0073] No NAG / wt% Succinic acid / wt% NAG to succinic acid mass ratio Leg-1 0.1％ - - Leg-2 - 0.1％ - Leg-3 0.0010％ 0.0990％ 0.01：1 Leg-4 0.0091％ 0.0909％ 0.1：1 Leg-5 0.0333％ 0.0667％ 0.5：1 Leg-6 0.050％ 0.050％ 1：1 Leg-7 0.0750％ 0.025％ 3：1 Leg-8 0.0909％ 0.0091％ 10：1 Leg-9 0.0978％ 0.0022％ 45：1 Leg-10 0.0980％ 0.0020％ 49：1

[0074] The NAG solution in Leg-1 was prepared by dissolving 0.1 g of NAG in 99.9 g of deionized water and stirring until completely dissolved.

[0075] The preparation method of succinic acid solution in Leg-2 is as follows: 0.1 g succinic acid is dissolved in 99.9 g deionized water and stirred until completely dissolved.

[0076] The NAG and succinic acid solutions in Leg-3 were prepared by dissolving 0.001 g of NAG and 0.099 g of succinic acid in 99.9 g of deionized water and stirring until completely dissolved.

[0077] The NAG and succinic acid solutions in Leg-4 were prepared by dissolving 0.0091 g of NAG and 0.0909 g of succinic acid in 99.9 g of deionized water and stirring until completely dissolved.

[0078] The NAG and succinic acid solutions in Leg-5 were prepared by dissolving 0.0333 g of NAG and 0.0667 g of succinic acid in 99.9 g of deionized water and stirring until completely dissolved.

[0079] The NAG and succinic acid solutions in Leg-6 were prepared by dissolving 0.05 g of NAG and 0.05 g of succinic acid in 99.9 g of deionized water and stirring until completely dissolved.

[0080] The NAG and succinic acid solutions in Leg-7 were prepared by dissolving 0.075 g of NAG and 0.025 g of succinic acid in 99.9 g of deionized water and stirring until completely dissolved.

[0081] The NAG and succinic acid solutions in Leg-8 were prepared by dissolving 0.0909 g of NAG and 0.0091 g of succinic acid in 99.9 g of deionized water and stirring until completely dissolved.

[0082] The NAG and succinic acid solutions in Leg-9 were prepared by dissolving 0.0978 g of NAG and 0.0022 g of succinic acid in 99.9 g of deionized water and stirring until completely dissolved.

[0083] The NAG and succinic acid solutions in Leg-10 were prepared by dissolving 0.098 g of NAG and 0.002 g of succinic acid in 99.9 g of deionized water and stirring until completely dissolved.

[0084] 2. Experimental methods

[0085] 2.1 Cellular melanin synthesis inhibition test method

[0086] Melanocytes in the logarithmic growth phase were collected and the cell density was 2×10 5 Each well was inoculated into a 24-well plate and incubated in an incubator (37°C, 5% CO 2 ) After culturing for 24 h, 1 mL of the experimental sample shown in Table 1 was added to each well, and 1 mL of kojic acid (3 mg / mL) was added as a positive control. Untreated cells were used as blank controls. Incubate in an incubator (37°C, 5% CO 2 ) for 24 h, discard the supernatant, add 0.5 mL of 1 mol NaOH containing 10% DMSO, and incubate in a constant temperature oven at 80°C for 1 h. After returning to room temperature, transfer 200 μL to each well of a 96-well plate, use 1 mol NaOH containing 10% DMSO as a zero control, read the absorbance at 405 nm, and calculate the cell melanin synthesis inhibition rate.

[0087]

[0088] 2.2 Cellular tyrosinase activity inhibition test method

[0089] Melanocytes in the logarithmic growth phase were collected and the cell density was 1×10 5 Inoculate 1000 cells / well into a 24-well plate. Incubate in an incubator (37°C, 5% CO 2 ) for 24 h, 1 mL of the experimental sample shown in Table 1 was added to each well, and 1 mL of kojic acid (3 mg / mL) was added as a positive control, and untreated cells were used as a blank control. Three parallels were set for each group. After adding the sample, the cells were placed in an incubator (37°C, 5% CO 2 ) for 24 h, discard the culture medium, wash twice with PBS, set the zeroing group at the same time, add only 100 μL NP-40 lysis buffer, lyse on ice for 5 min, take out the 24-well plate, aspirate 50 μL of supernatant and add it to a new 96-well plate, add 50 μL 1 mg / mL L-DOPA to each well, incubate at 37 °C for 1 h, then read the absorbance at 475 nm and calculate the cell tyrosinase activity inhibition rate.

[0090]

[0091] 3. Experimental results

[0092] The melanin synthesis inhibition rate and tyrosinase activity inhibition rate of cells after treatment with different experimental samples are shown in Table 2 below.

[0093] Table 2 Inhibition rate of cell melanin synthesis and cell tyrosinase activity of different experimental samples

[0094] Melanin synthesis inhibition rate Cellular tyrosinase activity inhibition rate Leg-1 33.20 23.25 Leg-2 37.53 24.19 Leg-3 37.13 23.57 Leg-4 29.36 23.06 Leg-5 44.82**# 28.51***## Leg-6 43.99**# 25.82*# Leg-7 44.07**# 26.12**# Leg-8 45.61**# 32.02***## Leg-9 43.14**# 27.78**# Leg-10 40.59 24.39

[0095] * indicates statistically significant difference compared with Leg-1 group, p<0.05;

[0096] ** indicates a statistically significant difference compared with the Leg-1 group, p < 0.01;

[0097] *** indicates statistically significant difference compared with Leg-1 group, p<0.001;

[0098] # indicates statistically significant difference compared with Leg-2 group, p<0.05;

[0099] ## indicates a statistically significant difference compared with the Leg-2 group, p<0.01.

[0100] The test results in Table 2 show the cell melanin synthesis inhibition rate and cell tyrosinase activity inhibition rate of samples with a concentration of 0.1% NAG, 0.1% succinic acid, and a total concentration of NAG and succinic acid maintained at 0.1%. The results show that compared with the use of NAG and succinic acid alone, when NAG and succinic acid are compounded in a ratio of 0.5-45:1, the cell melanin synthesis inhibition rate and the cell tyrosinase activity inhibition rate are significantly improved, that is, NAG and succinic acid have a synergistic effect on skin whitening in the ratio range of 0.5-45:1, with a statistically significant difference, proving that NAG and succinic acid have a synergistic whitening effect within this range.

[0101] Experimental Example 2: Corneal desmosin (CDSN) test of NAG, succinic acid and their combination based on epidermal model

[0102] This test establishes an epidermal model to test the expression of the intercellular junction desmosome protein CDSN, and compares the reduction in CDSN expression by single NAG, single succinic acid, and the combination of the two, indicating the efficacy of the two substances in synergistically softening keratin.

[0103] 1. Experimental samples

[0104] The contents of effective ingredients in the experimental samples are shown in Table 3 below.

[0105] Table 3 Composition and concentration of each experimental sample

[0106] No NAG / wt% Succinic acid / wt% Leg-1 1％ - Leg-2 - 1％ Leg-3 0.5％ 0.5％

[0107] The NAG solution in Leg-1 was prepared by dissolving 1 g of NAG in 99 g of deionized water and stirring until completely dissolved.

[0108] The preparation method of succinic acid solution in Leg-2 is as follows: dissolve 1 g of succinic acid in 99 g of deionized water and stir until completely dissolved.

[0109] The NAG and succinic acid solutions in Leg-3 were prepared by dissolving 0.5 g of NAG and 0.5 g of succinic acid in 99 g of deionized water and stirring until completely dissolved.

[0110] 2. Epidermal model corneal desmosin (CDSN) content test method

[0111] Prepare 1% NAG, 1% succinic acid and 0.5% NAG + 0.5% succinic acid solutions respectively and transfer them to 24-well plates (add 0.9 mL of the model culture medium of the corresponding group in advance). ), mark the test group number on the 24-well plate, incubate overnight, and add only 0.9 mL of model culture medium The three groups of samples and the blank control group were evenly smeared with 50 μL on the surface of the skin injury model, which was a finished 3D epidermal skin model purchased from Genevitra. When the experiment started, place it in a 24-well plate and add 0.9 mL Culture medium, placed in CO 2 Culture in an incubator (37±1℃, 5±1% CO2). Change the culture medium every day during the experiment. That's it;

[0112] After the drug administration, the model was placed in CO 2 Incubator (37°C, 5% CO 2) and continue incubation for 24h. Then, use a washing bottle filled with sterile PBS solution to clean the test substance remaining on the surface of the model, and use a sterile cotton swab to gently wipe off the residual liquid inside and outside the model. The skin model was removed by circumcision and fixed with 4% paraformaldehyde. After fixation, immunofluorescence operation was performed. The main steps are: dewaxing the paraffin sections to water, placing the sections in environmentally friendly dewaxing solution and washing them repeatedly for 3 times, 10min each time, then placing them in anhydrous ethanol and washing them repeatedly for 3 times, 5min each time, and finally washing with distilled water. During the antigen repair process, excessive evaporation of the buffer should be prevented, and the sections should not be dried. After the repair is completed, cool naturally. Place the slide in PBS (pHPH7.4) and shake it on a decolorizing shaker to wash 3 times, 5min each time. Block, after the sections are slightly dried, use a tissue pen to draw circles around the tissue, add BSA (the primary antibody is from goat and is blocked with 10% donkey serum, and the primary antibody from other sources is blocked with 3% BSA), and block for 30min. Add primary antibody: Add the prepared primary antibody dropwise, and incubate the slices flat in a humidified box at 4°C overnight. Add secondary antibody: Place the slides in PBS (pH7.4) and shake on a decolorizing shaker for 3 times, 5 minutes each time. Add the corresponding secondary antibody and incubate at room temperature in the dark for 50 minutes. DAPI counterstaining: Place the slides in PBS (pHPH7.4) and shake on a decolorizing shaker for 3 times, 5 minutes each time. Add DAPI staining solution and incubate at room temperature in the dark for 10 minutes. Quench tissue autofluorescence: Place the slides in PBS (pHPH7.4) and wash on a decolorizing shaker for 3 times, 5 minutes each time. Add autofluorescence quencher B solution for 5 minutes and rinse with running water for 10 minutes. Seal the slides with anti-fluorescence quenching sealant, then take pictures with a fluorescence microscope, and use ImageProPlus software to quantitatively analyze the CDSN fluorescence intensity.

[0113] 3. Experimental results

[0114] The expression of CDSN after treatment of different experimental samples is shown in Table 4 below.

[0115] Table 4 Differences in CDSN expression levels among different experimental samples

[0116] No Relative IOD fluorescence intensity average Leg-1 0.71 Leg-2 0.72 Leg-3 0.64*#

[0117] * indicates statistically significant difference compared with Leg-1 group, p<0.05;

[0118] # indicates a statistically significant difference compared with the Leg-2 group, p<0.05.

[0119] The results in Table 4 show that, compared with the use of NAG and succinic acid alone, the combination of NAG and succinic acid has the effect of significantly reducing the CDSN content, proving that the two can synergistically soften keratin.

[0120] Although the embodiments of the present invention are described above, the present invention is not limited to the above specific embodiments and application fields, and the above specific embodiments are only illustrative and instructive, but not restrictive. A person skilled in the art can make many forms under the guidance of this specification and without departing from the scope of protection of the claims of the present invention, all of which belong to the protection of the present invention.

Claims

1. A skin care composition, characterized in that: Contains acetyl glucosamine and succinic acid in a physiologically acceptable medium.

2. The skin care composition according to claim 1, characterized in that: The mass ratio of acetyl glucosamine to succinic acid is 0.01-50:

1.

3. The skin care composition according to claim 1, characterized in that: The mass ratio of acetyl glucosamine to succinic acid is 0.5-50:

1.

4. The skin care composition according to any one of claims 1 to 3, characterized in that: The skin care composition may further contain other active ingredients and / or auxiliary materials acceptable in the food or cosmetic field.

5. Use of a skin care composition containing acetyl glucosamine and succinic acid in the synergistic preparation of products for inhibiting tyrosinase activity and / or inhibiting melanin synthesis.

6. Use of a skin care composition containing acetyl glucosamine and succinic acid in synergistic skin whitening.

7. Use of the skin care composition according to claim 5 or 6, characterized in that: The mass ratio of the acetyl glucosamine to succinic acid is 0.5-50:

1.

8. Use of a skin care composition containing acetyl glucosamine and succinic acid in the synergistic preparation of a product for reducing corneal desmosin.

9. Use of a skin care composition containing acetyl glucosamine and succinic acid for synergistically softening skin keratin and / or exfoliating.

10. Use of the skin care composition according to claim 8 or 9, characterized in that: The mass ratio of the acetyl glucosamine to succinic acid is 0.5-50:1.

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