A polypeptide compound and its application in the field of cosmetics

Abstract

The present invention relates to the field of polymer applications, and in particular to a polypeptide compound and its application in the field of cosmetics. The present invention discloses a low molecular weight polypeptide compound, the structural formula of which is: [γ-glu] n ‑Aa, n is 1‑10; wherein glu is glutamic acid; Aa is selected from any one of glycine, alanine, valine, leucine, isoleucine, phenylalanine, tryptophan, tyrosine, aspartic acid, asparagine, glutamic acid, lysine, glutamine, methionine, serine, threonine, cysteine, proline, histidine or arginine. In the structural formula, n is preferably 1‑8. The molecular weight of the polypeptide derivative is 250‑1500Da. According to the test, the above polypeptide compound has excellent moisturizing performance and low molecular weight, which is conducive to absorption and utilization by the skin, reducing the risk of clogging pores and sweat gland openings.

Description

Technical Field

[0001] The present invention relates to the field of polymer applications, and in particular to a polypeptide compound and its application in the field of cosmetics. Background Art

[0002] Polyglutamic acid is a straight-chain linear polymer of homopolyglutamic acid connected by γ-amide bonds, which can be obtained by fermentation of Bacillus subtilis. The large number of peptide bonds on the main chain of polyglutamic acid are easily affected by light, heat and hydrolases in the environment, and degraded into non-toxic short peptides or glutamic acid monomers. Therefore, polyglutamic acid can be naturally degraded and is safe and environmentally friendly to use.

[0003] The structural formula of polyglutamic acid currently on the market is: [γ-glu] n , where n>200, and the molecular weight is around 1000000Da; due to its large molecular weight, when it is actually used as a skin care product, when it is applied to cosmetics and stays on the skin surface, especially in a humid environment or when sweating profusely, the polyglutamic acid will absorb moisture and swell, and may clog the skin sweat gland openings and pores, thereby causing local redness, swelling, pustules and other skin discomforts.

[0004] Therefore, polyglutamic acid needs to be properly degraded to prevent the above skin discomfort. The existing technology for the degradation of polyglutamic acid either has a complicated process or the performance of the product obtained by degradation is poor.

[0005] γ-Glutamyl peptides are a class of substances containing γ-glutamyl residues, which are generated by the dehydration condensation of the γ-carboxyl group of γ-glutamic acid and the amino group of amino acids or peptides. They are widely distributed in animals, plants and microorganisms in nature (Yao Yujing, Cui Chun, Huang Junwei. Research progress on the flavor characteristics, identification and preparation methods of γ-glutamyl peptides [J]. Chinese Condiments, 2016, 41(3): 146-152.). The study of γ-glutamyl peptides began in the 1950s. Some scholars separated and identified bean seed extracts and discovered a γ-glutamyl peptide (γ-L-glutamyl-S-methyl-L-cysteine) (Thompson J, Morris CJ, Zacharius R. Isolation of (-) S-methyl-L-cysteine ​​from beans(Phaseolus vulgaris)[J]. Nature, 1956, 178(4533): 593-593.). Later, naturally occurring γ-glutamyl peptides were found in legumes, mushrooms, alliums, tea, algae, and fermented condiments such as cheese, fish sauce, and soy sauce. γ-Glutamyl peptides are highly soluble in water and have health benefits such as anti-inflammatory, mood regulation, antioxidant, anti-epileptic, anti-cancer, memory improvement, and lowering blood sugar levels. The most representative of these is glutathione, a powerful antioxidant that protects the skin from oxidative damage and is a raw material for anti-aging skin care products (Puurunen J, Tiira K, Vapalahti K, et al. Fearful dogs have increased plasma glutamine and γ-glutamyl glutamine [J]. Scientific Reports, 2018, 8 (1): 1-12.). There are many types of γ-glutamyl peptides, but few have been developed as cosmetic raw materials. This study developed γ-glutamyl peptides into cosmetic raw materials with various functions.

[0006] The moisturizing or barrier repair function of the skin system is mainly affected by the following proteins:

[0007] Aquaporins, also known as water porins, are proteins located on the cell membrane that assist in the transport of water across the membrane. Water molecules can diffuse slowly across the membrane without the assistance of aquaporins, while aquaporins allow water molecules to flow out of or into the cell membrane quickly, helping cells adjust their internal osmotic pressure. The integumentary system includes skin, hair, nails, as well as nerves, fat, and some glands. In the integumentary system, AQP1, AQP3, AQP7, AQP9, and AQP10 are mainly expressed.

[0008] AQP3 is mainly expressed in keratinocytes and skin fibroblasts. Its effects on the skin are mainly in two aspects (Luo Ying. Research progress of water channel protein 3 and skin. Medical Information. 2015): First, moisturizing effect. Since the glycerol content in the stratum corneum directly or indirectly affects the moisturizing function of the skin, its source supply is extremely important. AQP3 can not only bring endogenous glycerol in the circulation and triglycerides in the sebaceous glands into the epidermis, but also further participate in the glycerol metabolism of epidermal cells; second, barrier repair function. When the skin barrier is damaged by artificial external force or chemical reagents, AQP3 expression increases, while intermediate filament-related protein (intermediate filament-related protein can provide natural moisturizing factor NMF after hydrolysis) shows a short and obvious decrease in expression 3 hours after barrier damage, and returns to normal in 24 hours. This is the peak of AQP3 expression increase. This shows that when intermediate filament-related protein decreases, AQP3 expression increases, replacing the role of intermediate filament-related protein, so the function of AQP3 in repairing the barrier may be achieved through interaction with intermediate filament-related protein.

[0009] Claudin-1 is also known as aging-associated epithelial membrane protein-1. Claudin-1 is located in the human chromosome 3q28 region, has a molecular weight of 22.7 kDa, and is composed of 211 amino acid residues. Claudin-1 contains four transmembrane domains, two extracellular loops ECL1 and ECL2, one cytoplasmic intracellular loop, and NH2- and COOH-terminals located in the cytoplasm. Adjacent cells are closed by the outer loop in a "zipper" structure. ECL1 is composed of approximately 50 amino acids, including a group of highly conserved residues W-GLWC-C. Its amino acid composition affects the paracellular charge selectivity and determines the properties of the paracellular pore. ECL1 contains two cysteines located at positions 54 and 64, respectively. These two cysteines form an intramolecular disulfide bridge, which is highly conservative and determines the binding behavior of the entire ECL1, which is crucial to the closing function of claudin-1. ECL2 is relatively small, containing 16 to 33 amino acid residues, mediating claudin-claudin interactions within cells and between adjacent cells, and acting as a barrier to the flow of extracellular water and solutes and the translocation of other cells by regulating the charge and size selection characteristics of the paracellular space. The C-terminus of Claudin-1 is a COOH-terminus ending in valine, with a length of 21 to 63 amino acid residues, containing a structural motif that binds to the PDZ domain, and can bind to zonula occludens-1, multi-PDZ domain protein 1, etc. to form tight junctions, ultimately achieving epithelial barrier function (Ji Rong et al., Progress in Research on the Function of Tight Junction Protein-1, Chinese Journal of Cell Biology, 2020).

[0010] Filaggrin (FLG) is an important protein that maintains the integrity of the epidermal barrier function. It is located on the human 1q21 locus and is encoded by the FLG gene. It is a key protein for normal epidermal terminal differentiation and maintaining skin hydration. Its precursor, profilaggrin (proFLG), is synthesized in the cells of the epidermal granular layer and participates in the formation of transparent keratin granules. When the calcium ion concentration increases, proFLG is dephosphorylated to become FLG, and is further hydrolyzed by serine proteases (SPs), and finally a large amount of NMF is generated under the action of caspase14. NMF is an important molecule that affects skin hydration. Its reduction or absence will weaken the skin barrier function and lead to the occurrence of various skin diseases.

[0011] In view of the problems existing in the prior art, targeted exploration is needed to determine suitable low molecular weight polypeptide compounds that meet the moisturizing and skin repair requirements of cosmetics. Summary of the invention

[0012] The first aspect of the present invention discloses a low molecular weight polypeptide compound, the structural formula of which is: [γ-glu] n -Aa, where glu is glutamic acid;

[0013] Aa is selected from any one of glycine, alanine, valine, leucine, isoleucine, phenylalanine, tryptophan, tyrosine, aspartic acid, asparagine, glutamic acid, lysine, glutamine, methionine, serine, threonine, cysteine, proline, histidine or arginine; n is 1-10.

[0014] Among them, Aa is preferably glutamine Gln.

[0015] In the structural formula, n is preferably 1-8, and n is selected from any number of 1, 2, 3, 4, 5, 6, 7 or 8.

[0016] The molecular weight of the polypeptide compound is 250-1500Da.

[0017] The polypeptide compound is prepared by a fermentation method.

[0018] The second aspect of the present invention discloses the use of the above-mentioned polypeptide compounds in moisturizing and skin barrier repair, including the use in the preparation of moisturizing products.

[0019] Preferably, the above polypeptide compounds are used in the preparation of moisturizing cosmetics, such as moisturizing masks, facial creams, lotions, etc.

[0020] The polypeptide compound is also used in the preparation of products for repairing skin barrier function. The products for repairing skin barrier function have firming and anti-wrinkle effects.

[0021] The third aspect of the present invention discloses the use of a polypeptide compound moisturizing product and one or a combination of the following substances:

[0022] I), an auxiliary agent that promotes the moisturizing effect of polypeptide compounds;

[0023] II) contains moisturizing agents that have a synergistic moisturizing effect.

[0024] The above-mentioned additives include antioxidants, preservatives, surfactants, pH adjusting buffers and other additives used in moisturizing products.

[0025] The above-mentioned moisturizers can respectively play the role of moisture absorption, water locking, bio-regulation or film-forming moisturizing, such as glycerin, propylene glycol, butylene glycol, mineral oil, vegetable oil, animal oil, ceramide, free fatty acids, triglycerides, squalene, sodium hyaluronate, pullulan, tremella polysaccharide, etc.

[0026] After testing, the above-mentioned polypeptide compounds have excellent moisturizing properties and low molecular weight, which is conducive to being absorbed and utilized by the skin, reducing the risk of clogging pores and sweat gland openings.

[0027] The polypeptide compound provided by the present invention has a molecular weight of 250-1500Da and can be used as a moisturizer in cosmetics to effectively moisturize, promote absorption, and increase skin elasticity. It can also be used as a drug carrier in the medical field, and has a low molecular weight, which can reduce the side effects caused by drugs. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 This is the secondary mass spectrum of the structure γ-Glu-Gln.

[0029] Figure 2 This is the secondary mass spectrum of the structure γ-Glu-Glu-Gln.

[0030] Figure 3 This is the secondary mass spectrum of the structure γ-Glu-Glu-Glu-Gln.

[0031] Figure 4 This is the secondary mass spectrum of the structure γ-Glu-Glu-Glu-Glu-Gln.

[0032] Figure 5 This is the secondary mass spectrum of the structure γ-Glu-Glu-Glu-Glu-Glu-Glu-Gln.

[0033] Figure 6 This is a diagram showing the moisturizing effect of the polypeptide compound disclosed in Example 2.

[0034] Figure 7 Bar graph showing the effects of peptides on HaCaT cell proliferation.

[0035] Figure 8 Bar graph showing the effect of oligomeric hyaluronic acid on the proliferation of HaCaT cells.

[0036] Fig. 9 Changes in AQP1 expression in HaCaT cells.

[0037] Fig.10 Changes in AQP3 expression in HaCaT cells.

[0038] Fig.11 Changes in FLG expression in HaCaT cells.

[0039] Fig.12 Changes in Claudin-1 expression in HaCaT cells.

[0040] Fig.13 This is a diagram showing the measurement results of the transepidermal water loss experiment in Example 4.

[0041] Fig.14 This is a diagram showing the measurement results of the skin elasticity experiment in Example 4.

[0042] Fig.15 This is a diagram showing the measurement results of the skin density experiment in Example 4.

[0043] Fig.16 This is a diagram showing the measurement results of the skin stratum corneum thickness experiment in Example 4.

[0044] Fig.17 This is a diagram showing the measurement results of the skin stratum corneum moisture experiment in Example 4.

[0045] Fig.18 This is a graph showing the measurement results of the cell activity experiment in Example 5.

[0046] Fig.19 This is a graph showing the expression of AQP1, AQP3, Claudin-1 and FLG genes after treatment with different polypeptide compounds in Example 6.

[0047] Wherein HA refers to oligomeric hyaluronic acid, and polypeptide refers to the polypeptide compound prepared in Example 1

[0048] The symbol * represents a statistically significant difference compared with the blank control group (control), * represents P < 0.05, ** represents P < 0.01, and *** represents P < 0.001; the symbol # represents a statistically significant difference between the two treatment groups, # represents P < 0.05, ## represents P < 0.01, and ### represents P < 0.001. DETAILED DESCRIPTION

[0049] The present invention is further described below by way of examples, but the present invention is not limited to the scope of the examples. The experimental methods in the following examples without specifying specific conditions are selected according to conventional methods and conditions, or according to the product specifications. The raw materials and equipment used in the examples are well known to those skilled in the art, and are all available on the market or easily obtained or prepared.

[0050] The experimental methods in the following examples, unless otherwise specified, are all conventional methods, and are performed according to the techniques or conditions described in the literature in the field or according to the product instructions. The materials, reagents, etc. used in the following examples, unless otherwise specified, can all be obtained from commercial channels.

[0051] Example 1: Mass spectrometry identification of samples

[0052] The present invention uses amino acids as raw materials and synthesizes polypeptide compounds by fermentation. The structure of the polypeptide compounds is determined by HPLC-MS / MS. Figure 1-5 shown.

[0053] Example 2: Zebrafish Moisturizing Efficacy Test

[0054] 1. System and sample size

[0055] Test system: zebrafish (Albino) with melanin allele mutation, sample solution: a mixed system containing the polypeptide compound disclosed in Structural Example 1.

[0056] Zebrafish age: 2 days post fertilization (2 dpf).

[0057] The number of experimental samples in each group was 15 (N=10). Normal control group: zebrafish were not dehydrated; model control group: zebrafish were dehydrated; sample group: zebrafish were immersed in sample solution for dehydration.

[0058] Adult fish feeding and breeding methods: in accordance with the company's laboratory standard feeding and breeding methods, in line with the requirements of the international AAALAC certification (certification number: 001458)

[0059] 2. Principle and method: When zebrafish are treated with sodium chloride, the skin surface will lose water and shrink due to osmotic pressure, and the tail area will become smaller due to shrinkage. The moisturizing effect of cosmetics is evaluated by the impact on the tail area.

[0060] 3. Experimental steps:

[0061] 1) Randomly select zebrafish and place them in a 6-well plate, with 15 zebrafish in each well.

[0062] 2) Water-soluble samples were administered, and a normal control group and a model control group were set up at the same time. The volume of each well was 3 mL.

[0063] 3) Sodium chloride was administered in water at the same time to establish a zebrafish skin dehydration model.

[0064] 4) Incubate at 28°C in dark for 22 h.

[0065] 5) Ten zebrafish were randomly selected from each experimental group and photographed under a dissecting microscope. Advanced image processing software was used to analyze and collect data. The tail area (S) of the zebrafish was analyzed and the moisturizing effect of the sample was calculated according to the formula to determine whether it had moisturizing effect. (Statistical analysis p<0.05 was considered to be significantly different.)

[0066]

[0067] 4. Test results

[0068] Test items Detection concentration (%) effect(%) p-value Test results Moisturizing effect 0.1 59 <0.05 Significant

[0069] The results are as follows Figure 6 As shown, it can be seen that the tail area of ​​the sample disclosed in Example 2 is significantly increased compared with the model control group, revealing that the sample has a moisturizing effect.

[0070] Example 3: Cell activity experiment

[0071] 1. Effect of peptides on the proliferation of human immortalized skin keratinocyte cell line HaCaT

[0072] Experimental method: HaCaT cells were revived within 1 min at 37°C in DMEM medium containing 10% fetal bovine serum and 1% triple antibody. They were cultured in an incubator at 37°C and 5% CO2, and passaged once every 48h-72h, with a passage ratio of 1:3. Cells in the exponential growth phase were collected, counted, and counted at 1×10 4 HaCaT cells were plated in a 96-well plate with 200 μL complete DMEM medium per well. After culturing for 24 hours, the cells were divided into a control group and an experimental group. The control group was added with 0.05%, 0.1%, 0.2%, 0.5% and 1% oligomeric hyaluronic acid, and the experimental group was added with 0.05%, 0.1%, 0.2%, 0.5% and 1% of the mixture of polypeptide compounds in Example 1. After 24 hours of sample addition, 10 μL CCK8 reagent was added to each well to detect cell proliferation ability.

[0073] Experimental results, such as Figure 7 , 8 As shown, compared with the control group, oligomeric hyaluronic acid had no significant effect on the proliferation of HaCaT cells at different concentrations, and the polypeptide compound mixture of Example 1 had no significant effect on the proliferation of HaCaT cells within the concentration range of 0-1.0%.

[0074] 2 peptides for the effect of AQP1, AQP3, Claudin-1 and FLG genes

[0075] Experimental methods: (1) Collect cells in the exponential growth phase, plate HaCaT cells in 12-well plates, and culture HaCaT cells for 24 hours using culture medium containing 0.05% and 0.5% oligomeric hyaluronic acid and 0.05% and 0.5% of the mixture of polypeptide compounds in Example 1, respectively, to detect the expression of water channel proteins AQP1 and AQP3, tight junction protein Claudin-1 and filaggrin FLG genes. (2) Culture at 37°C for 24 hours, and extract cell RNA using the TRIzol method. (3) Reverse transcribe RNA using a kit to obtain cDNA. (4) qPCR method was used to detect the expression of water channel proteins (AQP1, AQP3), tight junction protein (Claudin-1), and FLG genes in each group. qPCR reaction conditions: 95 ℃ 4 min, 95 ℃ 15 sec, 60 ℃ 30sec, 40 repeats, 95 ℃ 15 sec, 60 ℃ 1min, 37 ℃ 1min.

[0076] The experimental results, such as Fig. 9 , 10 As shown in Figures 11 and 12, after the cells were treated with 0.05% oligomeric hyaluronic acid, the expression levels of AQP1, AQP3, Claudin-1 and FLG did not change significantly. After the cells were treated with 0.5% oligomeric hyaluronic acid, the expression levels of AQP3, Claudin-1 and FLG increased significantly. After treatment with the mixture of polypeptide compounds in Example 1, the expression levels of AQP1, AQP3, Claudin-1 and FLG increased, and the expression levels of each gene increased more when the polypeptide was 0.5% compared with 0.05%. The results show that at the same detection amount, compared with oligomeric hyaluronic acid, the polypeptide has better moisturizing and barrier repair functions.

[0077] Example 4

[0078] [γ-glu] n -Gln human skin repair effect

[0079] Experimental methods:

[0080] Recruit 10 volunteers (aged 20-27 years old) and give them a 9cm diameter patch on the inner side of their forearms. 2 , sample concentration 1%, control 1: ceramide (Chongqing Zhihe); control 2: ceramide (Evonik); sample: [γ-glu] n -Gln.

[0081] Test method:

[0082] 1. Transepidermal water loss

[0083] The subjects were tested with the instrument four times, before use, 30 minutes after use, 7 days after use, and 14 days after use. The inner forearm area was selected for testing.

[0084] Each time you use the Tewameter test, read three values ​​and take the average.

[0085] Parameter explanation: The higher the test value, the higher the transepidermal water loss of the skin and the worse the skin barrier function. The lower the test value, the lower the transepidermal water loss of the skin and the better the skin barrier function.

[0086] Result determination: If after using the product, the transepidermal water loss in the test area is significantly reduced compared with the initial value before use, it means that the test sample has the effect of improving the skin barrier function.

[0087] 2. Skin elasticity

[0088] The subjects were tested with the instrument four times before use, 24 hours after use, 7 days after use, and 14 days after use. The inner area of ​​the right forearm was selected for testing.

[0089] Each time you use the Cutometer test, read three values ​​and take the average.

[0090] Parameter explanation: The R2 value is the total elasticity of the skin. Values ​​closer to 1 indicate higher elasticity.

[0091] Result determination: If the skin elasticity and collagen values ​​in the test area after use of the product are significantly increased compared with the initial values ​​before use, it means that the test sample has the effect of increasing skin elasticity and improving skin firmness.

[0092] 3. Skin density

[0093] The subjects were tested with the instrument four times, before use, 30 minutes after use, 7 days after use, and 14 days after use. The inner area of ​​the right forearm was selected for testing.

[0094] Each time the Ultrascan UC22 skin ultrasound instrument is used for testing, three values ​​are read and the average is taken.

[0095] Parameter explanation: The higher the test value, the higher the skin density; the lower the test value, the lower the skin density.

[0096] Result determination: If after using the product, the skin density of the test area is significantly increased compared with the initial value before use, it means that the test sample has the effect of improving the skin barrier function.

[0097] 4. Skin epidermal thickness

[0098] The subjects were tested with the instrument four times, before use, 30 minutes after use, 7 days after use, and 14 days after use. The inner area of ​​the right forearm was selected for testing.

[0099] Each time the Ultrascan UC22 skin ultrasound instrument is used for testing, three values ​​are read and the average is taken.

[0100] Parameter explanation: The higher the test value, the thicker the skin epidermis; the lower the test value, the thinner the skin epidermis.

[0101] Result determination: If after using the product, the epidermal thickness of the test area increases significantly compared with the initial value before use, it means that the test sample has the effect of improving the skin barrier function.

[0102] 5. Water content of the stratum corneum

[0103] The subjects were tested with the instrument four times, before use, 30 minutes after use, 7 days after use, and 14 days after use. The inner forearm area was selected for testing.

[0104] Each time you use the Corneometer test, read three values ​​and take the average.

[0105] Parameter explanation: The higher the test value, the higher the moisture content of the skin stratum corneum.

[0106] Result determination: If the moisture content of the stratum corneum in the test area after use of the product is significantly increased compared with the initial value before use, it means that the test sample has the effect of hydrating and moisturizing.

[0107] Test conclusion:

[0108] Using the test product ([γ-glu] n -Gln) 30 minutes later, the water content of the skin in the test area increased compared with before use, with an improvement rate of 4.4%; the transepidermal water loss value decreased compared with before use, with an improvement rate of 13.47%; the total elasticity of the skin in the test area improved to a certain extent, with an improvement rate of 2.05%. This shows that after using this product once, it has the effect of moisturizing and hydrating the skin.

[0109] With the test product ([γ-glu] n-Gln) 7 days later, the water content of the skin in the test area increased significantly compared with before use, with an improvement rate of 3.76%; the transepidermal water loss value decreased significantly compared with before use, with an improvement rate of 18.69%, and there was a significant difference; the total elasticity of the skin in the test area improved to a certain extent, with an improvement rate of 3.67%, and there was a significant difference; the skin density and stratum corneum thickness improved to a certain extent, with improvement rates of 6.90% and 7.39% respectively. This shows that the product has significant moisturizing and hydrating effects, elasticizing the skin, and has a certain barrier repair effect after using it for 7 days.

[0110] Using the test product ([γ-glu] n -Gln) 14 days later, the water content of the skin in the test area increased significantly compared with before use, with an improvement rate of 13.99%, and a very significant difference; the transepidermal water loss value decreased significantly compared with before use, with an improvement rate of 16.42%, and a significant difference; the total elasticity of the skin improved significantly, with an improvement rate of 5.01%, and a very significant difference; the glossiness of the skin improved significantly, with an improvement rate of 9.5%, and a significant difference; the skin density and stratum corneum thickness improved significantly, with improvement rates of 19.54% and 10.13%, respectively, and a significant difference. This shows that the product has significant moisturizing, hydrating, barrier repairing, and elastic skin effects after 14 days of use.

[0111] Example 5 [γ-glu] n -Aa cell viability assay

[0112] Experimental method: HaCaT cells were revived within 1 min at 37°C in DMEM medium containing 10% fetal bovine serum and 1% triple antibody. They were cultured in an incubator at 37°C and 5% CO2, and passaged once every 48h-72h, with a passage ratio of 1:3. Cells in the exponential growth phase were collected, counted, and counted at 1×10 4 HaCaT cells were plated in 96-well plates with 200 μL complete DMEM medium per well. After culturing for 24 h, the cells were divided into a control group and an experimental group. The control group was not treated with γ-glu n -Aa, experimental group added 0.001%, 0.005%, 0.01%, 0.05%, 0.1%, 0.2%, 0.5%, 1% and 2% [γ-glu] n -Aa. 24h after sample addition, 10μL CCK8 reagent was added to each well to detect cell proliferation ability.

[0113] The experimental results are shown in the figure, [γ-glu] n -Aa ([γ-glu] n -Trp, [γ-glu] n-Leu, [γ-glu] n -Phe, [γ-glu] n -Cys, [γ-glu] n -Met, [γ-glu] n -Tyr) had no significant effect on the proliferation of HaCaT in the concentration range of 0.001%~2.0%.

[0114] Example 6 [γ-glu] n -Aa effects on AQP1, AQP3, Claudin-1 and FLG genes

[0115] Experimental method: (1) cells in the exponential growth phase were collected and HaCaT cells were plated in 12-well plates. A control without sample, 0.05% and 0.2% [γ-glu] n -Trp, 0.05% and 1% [γ-glu] n -Leu, 1% and 2% [γ-glu] n -Phe, 0.01% and 0.05% [γ-glu] n -Cys, 0.01% and 0.2% [γ-glu] n -Met, 1% and 2% [γ-glu] n -Tyr medium was used to culture HaCaT cells for 24 h, and the expression of aquaporins AQP1 and AQP3, tight junction protein Claudin-1, and filaggrin FLG genes was detected. (2) After 24 h of culture at 37°C, cellular RNA was extracted using the TRIzol method. (3) RNA was reverse transcribed using a kit to obtain cDNA. (4) qPCR was used to detect the expression of aquaporins (AQP1, AQP3), tight junction protein (Claudin-1), and FLG genes in each group. qPCR reaction conditions: 95°C for 4 min, 95°C for 15 sec, 60°C for 30 sec, 40 repeats, 95°C for 15 sec, 60°C for 1 min, and 37°C for 1 min.

[0116] The experimental results are shown in the figure (1 and 2 refer to low and high concentrations respectively), 0.05% and 0.2% [γ-glu] n -Trp, 0.05% and 1% [γ-glu] n -Leu, 1% and 2% [γ-glu] n -Phe, 0.01% and 0.05% [γ-glu] n -Cys, 0.01% and 0.2% [γ-glu] n-Met, 1% and 2% [γ-glu] n After the cells were treated with -Tyr, the expression levels of AQP1, AQP3, Claudin-1 and FLG changed to varying degrees. n -Trp can significantly increase the expression of AQP3, Claudin-1 and FLG. 0.05% [γ-glu] n -Leu can significantly increase the expression of AQP1 and AQP3. 1% and 2% [γ-glu] n -Phe can significantly increase the expression of AQP1, AQP3, Claudin-1 and FLG. 2%[γ-glu] n -Tyr treatment significantly increased the expression of FLG. These results indicate that [γ-glu] n -Aa has moisturizing and barrier repair functions and can be used in various cosmetics.

[0117] [γ-glu] n -Aa in vitro efficacy study, [γ-glu] n -Gln in Gln can be replaced by other amino acids including but not limited to Trp, Leu, Phe, Cys, Met, Tyr and other 19 kinds of amino acids, through fermentation, enzymatic method or chemical synthesis [γ-glu] n -Aa, n is 1-9, Aa is an amino acid other than Gln. [γ-glu] n -Aa and [γ-glu] n -Gln, and can be used as a functional raw material in various cosmetics.

[0118] The above description is only a preferred specific implementation manner of the present invention, but the protection scope of the present invention is not limited thereto. Any technician familiar with the technical field can make equivalent replacements or changes according to the technical scheme and inventive concept of the present invention within the technical scope disclosed by the present invention, which should be covered by the protection scope of the present invention.

Claims

1. An application of a polypeptide compound in the field of cosmetics, characterized in that: Its structural formula is: [γ-glu] n -Gln, wherein glu is glutamic acid; Gln is glutamine; and n is 1-10.

2. The use according to claim 1, characterized in that In the structural formula, n is preferably 1-8.

3. The use according to claim 1, characterized in that The molecular weight of the polypeptide compound is 250-1500Da.

4. The use according to claim 1, characterized in that The polypeptide compound is prepared by a fermentation method.

5. The use according to claim 1, characterized in that Application of the polypeptide compound in the preparation of moisturizing products.

6. The use according to claim 1, characterized in that The polypeptide compound is used in the preparation of a product for repairing skin barrier function.

7. The use according to claim 5, characterized in that The moisturizing product includes the use of one or a combination of the following substances: I), an auxiliary agent that promotes the moisturizing effect of polypeptide compounds; II) contains moisturizing agents that have a synergistic moisturizing effect.

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