Active peptide, peptide-containing composition and use thereof

Abstract

The present invention relates to the field of biotechnology, and in particular to an active peptide, a peptide-containing composition and uses thereof. The present invention provides the use of collagen dipeptide and / or collagen tripeptide, a composition and uses thereof. Experiments show that the present invention can improve the moisture of the subject's skin, increase skin elasticity, reduce wrinkles, reduce melanin, significantly improve skin color, promote collagen synthesis, promote filaggrin synthesis, and promote hyaluronic acid synthesis.

Description

Technical Field

[0001] The present invention relates to the field of biotechnology, and in particular to an active peptide, a peptide-containing composition and use thereof. Background Art

[0002] As people age, their skin cells and tissues age and collagen is lost, which leads to the appearance of wrinkles and the appearance of surface aging. In the basic structure of the skin, the epidermis and dermis play an important role. Fibroblasts in the dermis can promote collagen synthesis. Collagen molecules cross each other to form fibers, forming a fine collagen fiber network in the skin, increasing the connection strength and stability of the skin. As an important component of human skin, collagen has a huge water-locking function, which directly determines the moisture, smoothness, firmness and skin age of the skin. In addition, the main function of the keratinocytes of the skin is to protect the subcutaneous tissue, maintain the normal skin barrier structure, and prevent excessive evaporation of water in the body. Hyaluronic acid can improve the integrity of the stratum corneum and repair damaged barriers.

[0003] The functions of common skin care products at present are based on whitening and moisturizing. The whitening and moisturizing effects are only for the epidermis of the skin. Some products that add hydrolyzed collagen, hyaluronic acid, peptides or retinol, etc., due to the large molecular weight of hydrolyzed collagen, it is not easy to penetrate the human skin barrier to reach the dermis, hyaluronic acid cannot essentially reduce the loss of collagen, and peptides and retinol have certain irritation and safety risks to the skin. They cannot penetrate into the dermis or subcutaneous tissue to improve the skin, repair the skin, promote skin metabolism, and tighten the skin. Existing skin care products lack the effect of promoting fibroblasts to secrete collagen and promoting the synthesis of hyaluronic acid in keratinocytes. Summary of the invention

[0004] In view of this, the present invention provides an active peptide, a peptide-containing composition and use thereof, which have the effects of whitening, firming and moisturizing.

[0005] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0006] The present invention provides the use of a peptide represented by formula (I) or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof in the preparation of a skin care product having whitening, firming and / or moisturizing effects.

[0007] Xaa1-Xaa2-Xaa3 (I)

[0008] Wherein, Xaa1 is an aliphatic amino acid, a polar amino acid or a neutral amino acid, Xaa2 is an amino acid containing a heterocyclic ring, a neutral amino acid or a non-polar amino acid; Xaa3 is any amino acid or none;

[0009] The whitening includes reducing skin melanin;

[0010] Said firming includes upregulating collagen content in cells;

[0011] The moisturizing includes up-regulating the content of filaggrin and / or the content of hyaluronic acid in cells.

[0012] In some specific embodiments of the present invention, the Xaa1 in the above application is glycine; and the Xaa2 is proline or hydroxyproline.

[0013] In some specific embodiments of the present invention, the peptide used above is collagen dipeptide and / or collagen tripeptide.

[0014] In some specific embodiments of the present invention, the peptide used above accounts for 0.2% to 2.0% of the net weight of the skin care product;

[0015] The collagen dipeptide accounts for 0.1% to 1.0% of the net weight of the skin care product;

[0016] The collagen tripeptide accounts for 0.1% to 1.0% of the net weight of the skin care product.

[0017] The present invention also provides a composition comprising collagen tripeptide and ectoine.

[0018] In some specific embodiments of the present invention, the above composition, by weight, comprises 0.1 to 1.0 parts of the collagen tripeptide and 0.1 to 2.0 parts of the ectoine.

[0019] The present invention also provides a composition comprising collagen tripeptide and sodium hyaluronate.

[0020] In some specific embodiments of the present invention, in the above composition, the collagen tripeptide is 0.1 to 1.0 part, and the sodium hyaluronate is 0.01 to 0.1 part, by weight.

[0021] The present invention also provides a composition comprising a peptide and sodium hyaluronate, wherein the peptide comprises a collagen dipeptide and a collagen tripeptide.

[0022] In some specific embodiments of the present invention, in the above composition, the collagen tripeptide is 0.1-1.0 part, the collagen dipeptide is 0.1-1.0 part, and the sodium hyaluronate is 0.01-0.1 part, by weight.

[0023] The present invention also provides the use of the above composition in preparing skin care products;

[0024] The skin care products include skin care products for whitening, firming and / or moisturizing;

[0025] The whitening includes reducing skin melanin;

[0026] Said firming includes upregulating collagen content in cells;

[0027] The moisturizing includes up-regulating the content of filaggrin and / or the content of hyaluronic acid in cells.

[0028] The present invention also provides a skin care product, comprising the above composition and acceptable auxiliary materials or additives.

[0029] In some specific embodiments of the present invention, the above-mentioned skin care product:

[0030] In the above composition, the collagen tripeptide accounts for 0.1wt% to 1.0wt% of the net weight of the skin care product, and the ectoine accounts for 0.1wt% to 2.0wt% of the net weight of the skin care product; or

[0031] In the above composition, the collagen tripeptide accounts for 0.1wt% to 1.0wt% of the net weight of the skin care product, and the sodium hyaluronate accounts for 0.01wt% to 0.1wt% of the net weight of the skin care product; or

[0032] In the above composition, the collagen tripeptide accounts for 0.1wt% to 1.0wt% of the net weight of the skin care product, the collagen dipeptide accounts for 0.1wt% to 1.0wt% of the net weight of the skin care product, and the sodium hyaluronate accounts for 0.01wt% to 0.1wt% of the net weight of the skin care product.

[0033] The present invention also provides a method for preparing the skin care product, comprising the step of mixing the above composition with acceptable auxiliary materials or additives.

[0034] The present invention provides a compound as described in formula (I),

[0035] Xaa1-Xaa2-Xaa3, formula (I)

[0036] Among them, Xaa1 is an aliphatic amino acid, a polar amino acid, or a neutral amino acid, especially glycine, Xaa2 is an amino acid containing a heterocyclic ring, a neutral amino acid, or a non-polar amino acid, especially proline or hydroxyproline, and Xaa3 is any amino acid or none.

[0037] In some specific embodiments of the present invention, the above-mentioned compound, wherein the amino acid modification in the compound comprises prolylation.

[0038] In some specific embodiments of the present invention, the above-mentioned compound, the amino acid sequence is Gly-Xaa2-Xaa3, its biologically active fragments, its salts, its combinations and / or its variants.

[0039] In some specific embodiments of the present invention, the above-mentioned compound has an average molecular weight of 280 to 600 Daltons.

[0040] The present invention provides a composition, comprising the above compound, wherein the at least one compound is present in the range of 0.5% to 1% of the total weight of the composition and at a concentration of 0.05 μM or more.

[0041] In some specific embodiments of the present invention, the above composition further comprises a cationic surfactant.

[0042] In some specific embodiments of the present invention, the above composition, wherein the cationic surfactant is present in the range of WT% of the total weight of the composition.

[0043] In some specific embodiments of the present invention, the above composition has a moisturizing effect of 20 minutes to 72 hours or longer, and has effective skin tightening and anti-wrinkle effects.

[0044] In some specific embodiments of the present invention, the above composition comprises an additional active ingredient selected from whitening agents, anti-redness agents, sunscreens and UV organic or inorganic filters, hydrating agents, moisturizing agents, wetting agents, scrubs, anti-wrinkle agents, anti-aging agents, slimming agents, anti-acne agents, anti-inflammatory agents, antioxidants, free radical scavengers, tanning agents, depilatories or shaving agents, hair growth slowing agents, tightening agents, peptides and vitamins.

[0045] In some specific embodiments of the present invention, the above-mentioned composition is used to prevent and / or treat the intrinsic and extrinsic symptoms of skin aging, enhance antioxidant effect, moisturize the skin, eliminate wrinkles, fine lines, skin breaks and improve skin roughness, enhance skin tension and soothe the skin, reduce skin sagging, skin spots and / or improve skin brightness.

[0046] In some embodiments of the present invention, the above composition is used to improve the mechanical properties of the skin, the tension and / or stiffness and / or elasticity of the skin.

[0047] The present invention provides a cosmetic method for treating wrinkles, comprising topically applying the above composition to the skin.

[0048] The active peptides, peptide-containing compositions and uses thereof of the present invention have the following effects:

[0049] 1. Experiments show that the spray water of the present invention can improve the moisture of the subject's skin, increase skin elasticity, and reduce wrinkles, indicating that the firming and anti-wrinkle effect of the present invention is relatively good;

[0050] 2. Experiments show that the essence of the present invention can significantly improve skin moisture and skin elasticity and improve wrinkles. At the same time, the combined use of collagen dipeptide and collagen tripeptide can more effectively improve skin condition and achieve a firming and anti-wrinkle effect;

[0051] 3. Experiments show that the whitening and anti-wrinkle spray of the present invention can reduce melanin, significantly improve skin color, and achieve effective whitening results;

[0052] 4. Experiments show that the content of type I collagen in fibroblasts treated with the present invention increases significantly, indicating that the present invention has the ability to promote fibroblasts to synthesize collagen;

[0053] 5. Experiments show that the content of filaggrin in keratinocytes significantly increases after application of the invention, indicating that the invention has the ability to promote the synthesis of filaggrin in keratinocytes;

[0054] 6. Experiments show that the hyaluronic acid content in keratinocytes significantly increases after application of the drug of the present invention, indicating that the drug of the present invention has the ability to promote the synthesis of hyaluronic acid in keratinocytes. BRIEF DESCRIPTION OF THE DRAWINGS

[0055] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art are briefly introduced below.

[0056] Figure 1 The figure shows the statistical graph of collagen content; "*" indicates significant difference compared with the blank control group, "***": p<0.001, "****": p<0.0001;

[0057] Figure 2 It shows the up-regulation rate of type Ⅰ collagen;

[0058] Figure 3 Statistical graph of filaggrin content; "*" indicates significant difference compared with the blank control group, "**": p<0.01, "****": p<0.0001;

[0059] Figure 4 The up-regulation rate of filaggrin is shown;

[0060] Figure 5 Shows the statistical graph of hyaluronic acid content; "*" indicates significant difference compared with the blank control group, "**": p<0.01;

[0061] Figure 6 Shows the upregulation rate of hyaluronic acid. DETAILED DESCRIPTION

[0062] The present invention discloses an active peptide, a peptide-containing composition and its use. Those skilled in the art can refer to the content of this article and appropriately improve the process parameters to achieve it. It should be particularly noted that all similar substitutions and modifications are obvious to those skilled in the art, and they are all considered to be included in the present invention. The method and application of the present invention have been described through preferred embodiments, and relevant personnel can obviously modify or appropriately change and combine the methods and applications described herein without departing from the content, spirit and scope of the present invention to implement and apply the technology of the present invention.

[0063] Unless otherwise specified, the raw materials, reagents, consumables and instruments involved in the present invention are all common commercial products and can be purchased from the market.

[0064] The present invention will be further described below in conjunction with embodiments:

[0065] Example 1

[0066] Firming anti-wrinkle spray, the formula is shown in Table 1, preparation method:

[0067] 1. Heat water, disodium EDTA, allantoin, glycerin and propylene glycol to 85 degrees and stir to dissolve evenly;

[0068] 2. Cool down to 45 degrees, add tripeptide-29 (collagen tripeptide, sequence is Gly-Pro-Hyp), Tremella polysaccharide, tetrahydromethylpyrimidine carboxylic acid, 1,2-hexanediol, and p-hydroxyacetophenone, and stir well.

[0069] Table 1

[0070]

[0071]

[0072] Comparative Example 1

[0073] The formula is shown in Table 2, and the preparation method is the same as Example 1.

[0074] Table 2

[0075] Serial number Raw material name percentage% 1 water TO 100 2 Disodium EDTA 0.02 3 Allantoin 0.20 4 glycerin 5.00 5 Propylene glycol 5.00 6 - - 7 Tremella polysaccharide 0.02 8 Tetrahydromethylpyrimidine carboxylic acid 0.50 9 1,2-Hexanediol 0.50 10 p-Hydroxyacetophenone 0.50

[0076] Effect Example 1

[0077] Experimental methods:

[0078] Thirty subjects (18 to 50 years old), both male and female, were selected and numbered separately.

[0079] Preparation before the test: No products (cosmetics or topical medicines) can be used on the test area 2 to 3 days before the test, and the test area cannot come into contact with water for 1h to 3h. Before the test, the subjects need to clean their faces and necks uniformly and wipe them clean with dry facial tissues. After cleaning, mark the measurement areas on the face and neck of the subjects. Before the formal test, you should sit quietly in a standard room for 30 minutes, without drinking water, and be placed in the test state and keep relaxed. In the experiment, a 2cm*2cm test area was marked on the face and neck, and multiple areas were marked on the face and neck at the same time, with an interval of 1cm between the areas. Multiple areas are evenly distributed on the face and neck. The subjects used an aqueous solution without active ingredients, a spray containing a firming and anti-wrinkle efficacy composition prepared in Example 1 in the experiment, and a spray prepared in Comparative Example 1, respectively, where water was used as a blank control group, and the test sample was measured at (3.0±0.1) mg / cm 2 Use a latex finger cot to evenly apply the test product to the test area, and use each sample in the morning and evening for one week respectively.

[0080] After one week, the 20 subjects were scored for different test items (0-100 points) in terms of skin hydration, skin elasticity and wrinkle changes, as shown in Table 3.

[0081] Table 3: Scoring criteria

[0082] Test items Scoring Criteria Hydration Good hydration: 71-100; Average hydration: 41-70; Poor hydration: 0-40 Skin elasticity Good skin elasticity 80-100; Average skin elasticity 51-79; No change in skin elasticity 0-50 Wrinkle changes Wrinkles become significantly lighter 80-100; wrinkles become generally lighter 51-79; wrinkles remain unchanged 0-50;

[0083] Through the test subjects' scores on skin hydration, skin elasticity, and wrinkle changes, it can be seen that the spray water of Example 1 has a better firming and anti-wrinkle effect. The experimental results are shown in Table 4.

[0084] Table 4: Experimental results

[0085] Grouping Hydration Skin elasticity Wrinkle changes Blank control group 66 50 47 Comparative Example 1 72 60 50 Example 1 81 89 86

[0086] Example 2

[0087] Firming anti-wrinkle essence, the formula is shown in Table 5, preparation method:

[0088] 1. Heat water, xanthan gum, carbomer, allantoin, glycerin and propylene glycol to 85 degrees and stir to dissolve evenly;

[0089] 2. Cool down to 45 degrees, add tripeptide-29 (collagen tripeptide, sequence is Gly-Pro-Hyp), sodium hyaluronate, p-hydroxyacetophenone, 1,2-hexanediol, triethanolamine, and essence, and stir well.

[0090] Table 5

[0091]

[0092]

[0093] Example 3

[0094] The formula of the firming and anti-wrinkle essence is shown in Table 6, and the preparation method is the same as that of Example 2.

[0095] Table 6

[0096] Serial number Raw material name percentage% 1 water TO 100 2 Xanthan gum 0.20 3 Carbomer 0.12 4 Allantoin 0.20 5 glycerin 5.00 6 Propylene glycol 5.00 7 Collagen tripeptide (sequence: Gly-Pro-Hyp) 0.30 8 Collagen dipeptide (sequence: Gly-Pro) 0.20 9 Sodium Hyaluronate 0.03 10 p-Hydroxyacetophenone 0.50 11 1,2-Hexanediol 0.50 12 Triethanolamine 0.12

[0097] Effect Example 2

[0098] Experimental methods:

[0099] The test method was the same as that of Effect Example 1, and 40 subjects (18 to 50 years old) were selected, both male and female, and numbered separately.

[0100] Preparation before the test: No products (cosmetics or topical medicines) can be used on the test site 2 to 3 days before the test, and the test site cannot come into contact with water for 1h to 3h. Before the test, the subjects need to clean their faces and necks uniformly and wipe them clean with dry tissue paper. After cleaning, mark the measurement areas on the face and neck of the subjects. Before the formal test, you should sit quietly in a standard room for 30 minutes, cannot drink water, and be placed in the test state to keep relaxed. In the experiment, a 2cm*2cm test area was marked on the face and neck, and multiple areas were marked on the face and neck at the same time, with an interval of 1cm. Multiple areas are evenly distributed on the face and neck. The subjects were smeared with an aqueous solution containing no active ingredients (the active ingredients are collagen tripeptide, collagen dipeptide, and sodium hyaluronate) as a blank control group. In the experiment, the essence containing the firming and anti-wrinkle effect group prepared in Examples 2 to 3 was tested at (3.0±0.1) mg / cm 2 Use a latex finger cot to evenly apply the test product to the test area, and apply each sample in the morning and evening for one week respectively.

[0101] After application, the skin moisture, skin elasticity and wrinkle changes of 40 subjects were scored for different test items (0-100 points) as shown in Table 7.

[0102] Table 7: Scoring criteria

[0103] Test items Scoring Criteria Hydration Good hydration: 71-100; Average hydration: 41-70; Poor hydration: 0-40 Skin elasticity Good skin elasticity 80-100; Average skin elasticity 51-79; No change in skin elasticity 0-50 Wrinkle changes Wrinkles become significantly lighter 80-100; wrinkles become generally lighter 51-79; wrinkles remain unchanged 0-50;

[0104] Table 8: Experimental results

[0105] Grouping Hydration Skin elasticity Wrinkle changes Blank control group 62 55 45 Example 2 80 88 86 Example 3 89 95 93

[0106] Through the test subjects' scores on skin hydration, skin elasticity, and wrinkle changes, it can be compared that the essence water of the embodiment has a better firming and anti-wrinkle effect. According to the scoring results of the embodiment and the blank control group in Table 8, the wrinkle changes and skin elasticity of the skin added with firming and anti-wrinkle functional raw materials in Embodiments 2 and 3 are higher than those of the blank control group, and the skin hydration and skin elasticity can be significantly improved and wrinkles can be improved in Embodiment 3, indicating that the combined use of the raw materials in Embodiment 3 can more effectively improve the skin condition and achieve a firming and anti-wrinkle effect.

[0107] Example 4

[0108] Collagen tripeptide firming and anti-wrinkle efficacy test (in vitro fibroblast type I collagen content determination)

[0109] (1) Experimental methods

[0110] The experiment was designed according to the standard of "T / SHRH 031-2020 Cosmetic Firming and Anti-wrinkle Efficacy Test-Determination of Type I Collagen Content in Fibroblasts in Vitro", and the firming effect of the sample was verified through the sample group, blank control group and positive control group. After the fibroblasts were inoculated in a 96-well plate, they were given different treatments, without adding sample solution, adding sample solutions of different concentrations and stimulating factors for a period of incubation, and then the supernatant after culture was collected and tested using the instruction manual of the type I collagen enzyme-linked immunosorbent assay kit, and the absorbance value of each sample well was measured at 450nm using an enzyme reader.

[0111] (2) Correlation between experimental indicators and firming effects

[0112] Increase rate (%) = (AB) / B*100%

[0113] A: (sample solution / TGF-β1) OD value of sample solution;

[0114] B: OD value of blank control group solution;

[0115] The results of this experiment show that the product promotes the production of type I collagen. If the test sample is upregulated compared with the model blank control, and the improvement of the test product is significantly different from the model blank control, it means that the test sample has a firming effect. The significant difference is statistically analyzed by SPSS software. When the significance factor P<0.05, it is considered to be significantly different.

[0116] (3) Experimental results

[0117] Under normal culture conditions, the test substances affect the content and upregulation rate of collagen secreted by fibroblasts as shown in Table 9. Compared with the blank group, the content of type I collagen in the fibroblasts treated with the sample group increased significantly, and the collagen upregulation in the group containing both collagen tripeptide and sodium hyaluronate was higher, with an upregulation rate of 135.58% to 195.61%.

[0118] Table 9: Collagen content and upregulation rate in fibroblasts

[0119]

[0120] “*” indicates significant difference compared with the blank control group, “***”: p<0.001, “****”: p<0.0001.

[0121] (4) Experimental conclusion

[0122] Compared with the blank control, the content of type I collagen in fibroblasts of the sample group containing both collagen tripeptide and sodium hyaluronate increased significantly, with an increase rate of 135.58% to 195.61%, indicating that tripeptide-29 (collagen tripeptide) has the ability to promote fibroblasts to synthesize collagen. According to the experimental basis and the mechanism of skin aging, this result can be used as one of the evidences to support the firming effect claim of tripeptide-29.

[0123] Example 5

[0124] Collagen tripeptide moisturizing efficacy test (determination of the ability to promote hyaluronic acid synthesis in keratinocytes)

[0125] (1) Experimental methods

[0126] The experiment was designed according to the standards of the literature "Evaluation Specifications for Cosmetic Efficacy Claims" and "Effects of Ophiopogon japonicus and Its Compound Extracts on the Expression of Genes Related to Skin Moisturizing". The moisturizing effect of the product was verified through the sample group, blank control group, and positive control group. After keratinocytes were inoculated in a 96-well plate and cultured for 24 hours, they were given different treatments, without adding sample solution, and after adding different sample solutions for 24 hours, the contents of human filaggrin (FLG) and human hyaluronic acid (HA) were detected by the kit, and finally the absorbance of each sample well was measured at 450nm using an ELISA reader.

[0127] (2) Correlation between experimental indicators and moisturizing efficacy

[0128] Increase rate (%) = (AB) / B*100%

[0129] A: OD value of sample solutions with different concentrations;

[0130] B: OD value of blank control group solution;

[0131] The results of this experiment show that collagen tripeptide can promote the production of FLG and HA. If the expression levels of FLG and HA in the test sample are increased compared with the control group, and there is a significant difference between the test product and the model control, it means that the test sample has moisturizing ability. The significant difference is statistically analyzed by SPSS software. When the significance factor P<0.05, it is considered to be significantly different.

[0132] (3) Experimental results

[0133] ① Under normal culture conditions, the content and up-regulation rate of filaggrin in keratinocytes are shown in Table 10. Compared with the blank control group, the expression of filaggrin in keratinocytes treated with the sample was significantly up-regulated, and the combined use of collagen tripeptide and ectoine was more effective, with the up-regulation rate of filaggrin being 88.98% to 96.85%.

[0134] ② Under normal culture conditions, the hyaluronic acid content and up-regulation rate in keratinocytes are shown in Table 11. Compared with the blank control, the hyaluronic acid content in keratinocytes treated with the sample was significantly up-regulated, and the up-regulation rate was 87.65% to 97.2%.

[0135] Table 10: Filaggrin content and upregulation rate in human keratinocytes

[0136]

[0137]

[0138] “*” indicates significant difference compared with the blank control group, “**”: p<0.01, “****”: p<0.0001.

[0139] Table 11: Hyaluronic acid content and upregulation rate in keratinocytes

[0140]

[0141] “*” indicates significant difference compared with the blank control group, “**”: p<0.01.

[0142] (4) Experimental conclusion

[0143] Compared with the blank control, the keratinocyte filaggrin content and hyaluronic acid in the sample group containing collagen tripeptide and ectoine carboxylic acid increased significantly, indicating that collagen tripeptide and ectoine carboxylic acid have the ability to promote the synthesis of keratinocyte filaggrin, and the keratinocyte hyaluronic acid content in the 1% collagen tripeptide and 1% ectoine carboxylic acid sample group increased significantly, with an increase rate of 97.2%, indicating that collagen tripeptide and ectoine carboxylic acid have the ability to promote the synthesis of keratinocyte hyaluronic acid. According to the experimental basis and the skin moisturizing mechanism, this result can be used as one of the evidences to support the moisturizing effect claim of collagen tripeptide raw materials.

[0144] Example 6

[0145] Firming and anti-wrinkle efficacy test (in vitro fibroblast type I collagen content determination)

[0146] (1) Experimental methods

[0147] The experiment was designed according to the standard of "T / SHRH 031-2020 Cosmetic Firming and Anti-wrinkle Efficacy Test-Determination of Type I Collagen Content in Fibroblasts in Vitro", and different sample groups, blank control group and positive control group were tested to verify the firming effect of the samples. After the fibroblasts were inoculated in a 96-well plate, they were given different treatments, without adding sample solution, adding different sample solutions and stimulating factors, and incubated for a period of time. The supernatant after culture was collected and tested using the instruction manual of the type I collagen enzyme-linked immunosorbent assay kit, and the absorbance value of each sample well was measured at 450nm using an enzyme reader.

[0148] (2) Correlation between experimental indicators and firming effects

[0149] Increase rate (%) = (AB) / B*100%

[0150] A: (sample solution / TGF-β1) OD value of sample solution;

[0151] B: OD value of blank control group solution;

[0152] The experimental results show that the sample group promotes the production of type I collagen. If the test sample is upregulated compared with the blank control group, and the improvement of the test product is significantly different from the blank control, it means that the test sample has a firming effect. The significant difference is statistically analyzed by SPSS software. When the significance factor P<0.05, it is considered to be significantly different.

[0153] (3) Experimental results

[0154] Under normal culture conditions, the test substances affect the content and up-regulation rate of collagen secretion by fibroblasts as shown in Table 12. Compared with the blank group, the content of type I collagen in fibroblasts treated with the sample groups containing collagen tripeptide, collagen dipeptide and sodium hyaluronate increased significantly, with an up-regulation rate of 185.91% to 223.9%.

[0155] Table 12: Collagen content and upregulation rate in fibroblasts

[0156]

[0157]

[0158] “*” indicates significant difference compared with the blank control group, “***”: p<0.001, “****”: p<0.0001.

[0159] (4) Experimental conclusion

[0160] Compared with the blank control, the content of type I collagen in fibroblasts of the sample group containing collagen tripeptide, collagen dipeptide and sodium hyaluronate increased significantly, with an increase rate of 185.91% to 223.9%, indicating that tripeptide-29 (collagen tripeptide) has the ability to promote fibroblasts to synthesize collagen. According to the experimental basis and the mechanism of skin aging, this result can be used as one of the evidences to support the firming effect claim of tripeptide-29.

[0161] Example 7

[0162] Collagen tripeptide firming and anti-wrinkle efficacy test (in vitro fibroblast type I collagen content determination)

[0163] (1) Experimental methods

[0164] The experiment was designed according to the standard of "T / SHRH 031-2020 Cosmetic Firming and Anti-wrinkle Efficacy Test-Determination of Type I Collagen Content in Fibroblasts in Vitro", and the firming effect of the sample was verified through the sample group, blank control group and positive control group. After the fibroblasts were inoculated in a 96-well plate, they were given different treatments, without adding sample solution, adding sample solutions of different concentrations and stimulating factors for a period of incubation, and then the supernatant after culture was collected and tested using the instruction manual of the type I collagen enzyme-linked immunosorbent assay kit, and the absorbance value of each sample well was measured at 450nm using an enzyme reader.

[0165] (2) Correlation between experimental indicators and firming effects

[0166] Increase rate (%) = (AB) / B*100%

[0167] A: (sample solution / TGF-β1) OD value of sample solution;

[0168] B: OD value of blank control group solution;

[0169] The results of this experiment show that the product promotes the production of type I collagen. If the test sample is upregulated compared with the model blank control, and the improvement of the test product is significantly different from the model blank control, it means that the test sample has a firming effect. The significant difference is statistically analyzed by SPSS software. When the significance factor P<0.05, it is considered to be significantly different.

[0170] (3) Experimental results

[0171] Under normal culture conditions, the test substances affect the collagen secretion content and upregulation rate of fibroblasts as shown in Table 13. Figure 1 and Figure 2 Compared with the blank group, the content of type I collagen in fibroblasts in the 0.01% and 0.001% sample groups increased significantly, with the upregulation rates being 147.877% and 35.935% respectively.

[0172] Table 13: Collagen content and upregulation rate in fibroblasts

[0173]

[0174] “*” indicates significant difference compared with the blank control group, “***”: p<0.001, “****”: p<0.0001.

[0175] (4) Experimental conclusion

[0176] Compared with the blank control, the content of type I collagen in fibroblasts in the 0.01% and 0.001% sample groups increased significantly, with an increase rate of 147.877% and 35.935%, indicating that tripeptide-29 (collagen tripeptide) has the ability to promote fibroblasts to synthesize collagen. According to the experimental basis and the mechanism of skin aging, this result can be used as one of the evidences to support the firming effect claim of tripeptide-29.

[0177] Example 8

[0178] Collagen tripeptide moisturizing efficacy test (determination of the ability to promote hyaluronic acid synthesis in keratinocytes)

[0179] (1) Experimental methods

[0180] The experiment was designed according to the standards of the literature "Evaluation Specifications for Cosmetic Efficacy Claims" and "Effects of Ophiopogon japonicus and Its Compound Extracts on the Expression of Genes Related to Skin Moisturizing". The moisturizing effect of the product was verified through the sample group, blank control group, and positive control group. After keratinocytes were inoculated in a 96-well plate and cultured for 24 hours, they were given different treatments, without adding sample solution, and adding sample solutions of different concentrations for 24 hours. The contents of the two were detected by the human filaggrin (FLG) kit and the human hyaluronic acid (HA) kit, respectively, and finally the absorbance of each sample well was measured at 450nm using an ELISA reader.

[0181] (2) Correlation between experimental indicators and moisturizing efficacy

[0182] Increase rate (%) = (AB) / B*100%

[0183] A: OD value of sample solutions with different concentrations;

[0184] B: OD value of blank control group solution;

[0185] The experimental results are characterized by the production of FLG and HA from collagen tripeptide. If the expression of FLG and HA in the test sample is increased compared with the model control, and there is a significant difference between the test product and the model control, it means that the test sample has moisturizing ability. The significant difference is statistically analyzed by SPSS software. When the significance factor P<0.05, it is considered to be significantly different.

[0186] (3) Experimental results

[0187] ① Under normal culture conditions, the content and up-regulation rate of filaggrin in keratinocytes are shown in Table 14. Figure 3 and Figure 4 Compared with the blank control group, the content of filaggrin in keratinocytes in the 0.1% and 0.01% sample groups increased significantly, with the upregulation rates being 95.993% and 38.561%.

[0188] ② Under normal culture conditions, the hyaluronic acid content and upregulation rate in keratinocytes are shown in Table 15. Figure 5 and Figure 6 Compared with the blank control, the hyaluronic acid content in keratinocytes of the 0.1% and 0.01% sample groups increased significantly, with the upregulation rates being 65.055 and 21.422%, respectively.

[0189] Table 14: Filaggrin content and upregulation rate in human keratinocytes

[0190]

[0191] “*” indicates significant difference compared with the blank control group, “**”: p<0.01, “****”: p<0.0001.

[0192] Table 15: Hyaluronic acid content and upregulation rate in keratinocytes

[0193]

[0194] “*” indicates significant difference compared with the blank control group, “****”: p<0.0001, “*”: p<0.05.

[0195] (4) Experimental conclusion

[0196] Compared with the blank control, the keratinocyte filaggrin content of the 0.1% and 0.01% sample groups increased significantly, with an increase rate of 95.993% and 38.561%, indicating that collagen tripeptide has the ability to promote the synthesis of keratinocyte filaggrin, and the keratinocyte hyaluronic acid content of the 0.1% sample group increased significantly, with an increase rate of 65.055%, indicating that collagen tripeptide has the ability to promote the synthesis of keratinocyte hyaluronic acid. According to the experimental basis and the mechanism of skin moisturizing, this result can be used as one of the evidences to support the moisturizing effect claim of collagen tripeptide raw materials.

[0197] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. Use of a peptide represented by formula (I) or a stereoisomer thereof or a pharmaceutically acceptable salt thereof in the preparation of a skin care product having whitening, firming and / or moisturizing effects, Xaa1-Xaa2-Xaa3 (I) in, Xaa1 is an aliphatic amino acid, a polar amino acid or a neutral amino acid, Xaa2 is an amino acid containing a heterocyclic ring, a neutral amino acid or a non-polar amino acid; Xaa3 is any amino acid or none; The whitening includes reducing skin melanin; Said firming includes upregulating collagen content in cells; The moisturizing includes up-regulating the content of filaggrin and / or the content of hyaluronic acid in cells.

2. The use according to claim 1, characterized in that The Xaa1 is glycine; the Xaa2 is proline or hydroxyproline.

3. The use according to claim 1, characterized in that The peptides are collagen dipeptides and / or collagen tripeptides.

4. The use according to claim 1, characterized in that The peptide accounts for 0.2% to 2.0% of the net weight of the skin care product; The collagen dipeptide accounts for 0.1% to 1.0% of the net weight of the skin care product; The collagen tripeptide accounts for 0.1% to 1.0% of the net weight of the skin care product.

5. A composition, characterized in that Includes collagen tripeptide and ectoine.

6. The composition according to claim 5, characterized in that In parts by weight, the collagen tripeptide is 0.1 to 1.0 parts, and the ectoine is 0.1 to 2.0 parts.

7. A composition, characterized in that Includes collagen tripeptide and sodium hyaluronate.

8. The composition according to claim 7, characterized in that In parts by weight, the collagen tripeptide is 0.1 to 1.0 part, and the sodium hyaluronate is 0.01 to 0.1 part.

9. A composition, characterized in that It comprises peptides and sodium hyaluronate, wherein the peptides comprise collagen dipeptide and collagen tripeptide.

10. The composition according to claim 9, characterized in that In parts by weight, the collagen tripeptide is 0.1 to 1.0 part, the collagen dipeptide is 0.1 to 1.0 part, and the sodium hyaluronate is 0.01 to 0.1 part.

11. Use of the composition according to any one of claims 5 to 10 in preparing skin care products; The skin care products include skin care products for whitening, firming and / or moisturizing; The whitening includes reducing skin melanin; Said firming includes upregulating collagen content in cells; The moisturizing includes up-regulating the content of filaggrin and / or the content of hyaluronic acid in cells.

12. A skin care product, characterized in that: The composition comprises the composition according to any one of claims 5 to 10, and acceptable adjuvants or auxiliary agents.

13. The skin care product according to claim 12, characterized in that: include: In the composition according to claim 5 or 6, the collagen tripeptide accounts for 0.1wt% to 1.0wt% of the net weight of the skin care product, and the ectoine accounts for 0.1wt% to 2.0wt% of the net weight of the skin care product; or In the composition according to claim 7 or 8, the collagen tripeptide accounts for 0.1wt% to 1.0wt% of the net weight of the skin care product, and the sodium hyaluronate accounts for 0.01wt% to 0.1wt% of the net weight of the skin care product; or In the composition according to claim 9 or 10, the collagen tripeptide accounts for 0.1wt% to 1.0wt% of the net weight of the skin care product, the collagen dipeptide accounts for 0.1wt% to 1.0wt% of the net weight of the skin care product, and the sodium hyaluronate accounts for 0.01wt% to 0.1wt% of the net weight of the skin care product.

14. A method for preparing a skin care product, characterized in that: The method comprises the step of mixing the composition according to any one of claims 5 to 10 with an acceptable auxiliary material or adjuvant.

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