A blue copper-containing peptide-containing composition for repairing photothermal damage and application thereof

Abstract

This invention relates to the field of cosmetics, specifically to a composition containing copper peptides for repairing photothermal damage and its application. The composition comprises a complex polypeptide, galactosomal yeast fermentation product filtrate, and cyanobacterial extract; wherein the complex polypeptide includes copper peptides, yeast polypeptides, and hexapeptide-11. This invention, by combining specific complex polypeptides with galactosomal yeast fermentation product filtrate and cyanobacterial extract, increases the expression level of heat shock proteins, thereby achieving excellent efficacy in repairing photothermal damage. Furthermore, this composition is mild and non-irritating, and can be widely used in cosmetics.

Description

Technical Field

[0001] This invention relates to the field of cosmetic technology, and more particularly to a composition containing blue copper peptides for repairing photothermal damage and its application. Background Technology

[0002] In the cosmetics industry, photothermal damage to the skin (such as ultraviolet radiation, high-temperature environments, or medical aesthetic phototherapy) can lead to denaturation of skin cell proteins, damage to cell membrane structure, and release of inflammatory factors, thereby causing problems such as impaired skin barrier function, increased moisture loss, and sensitivity. When photothermal damage occurs, skin cells activate the stress expression mechanism of heat shock proteins (HSPs) to repair damaged proteins and maintain cellular homeostasis. However, excessive or prolonged photothermal damage may exceed the cell's own regulatory capacity of HSPs, resulting in insufficient repair efficiency and even triggering chronic inflammation or accelerating skin aging.

[0003] Currently, most repair products on the market focus on single-dimensional soothing or moisturizing, lacking a systematic solution that regulates HSP expression levels to achieve photothermal damage repair. For example, some products rely on single peptide ingredients to promote cell regeneration or passively defend against photothermal damage using mineral sunscreens, but fail to effectively activate the skin's endogenous protective mechanisms. Therefore, there is an urgent need to develop a composition that can actively enhance HSP expression levels to repair photothermal damage. Summary of the Invention

[0004] To address the aforementioned technical problems, the present invention aims to provide a composition containing blue copper peptides for repairing photothermal damage and its application, thereby overcoming the shortcomings of the prior art.

[0005] To achieve the above-mentioned technical effects, the present invention adopts the following technical solution:

[0006] In a first aspect, the present invention provides a composition containing copper peptides for repairing photothermal damage, the composition comprising a complex polypeptide, galactosomal yeast fermentation product filtrate, and cyanobacteria extract.

[0007] Preferably, the composite polypeptide comprises blue copper peptide, yeast polypeptide, and hexapeptide-11 in a mass ratio of 1:0.3-0.5:0.5-0.7.

[0008] Preferably, the mass ratio of the complex polypeptide, galactosomal yeast fermentation product filtrate, and cyanobacterial extract is 1:6-8:4-6.

[0009] Preferably, the cyanobacterial raw materials in the cyanobacterial extract include at least one of Suizenji cyanobacteria, Spirulina platensis, Nostoc, Oscillatoria, and Cyclocarya pallida.

[0010] Preferably, the cyanobacteria raw materials in the cyanobacteria extract include Suizenji cyanobacteria powder, Spirulina platensis powder, and Nostoc commune powder in a mass ratio of 1:0.5-1.5:3-5.

[0011] Preferably, the preparation method of the cyanobacteria extract includes the following steps:

[0012] A1. Mix Suizenji blue algae powder, Spirulina platensis powder, and Nostoc commune powder evenly in a mass ratio of 1:0.5-1.5:3-5 to obtain blue algae raw material powder;

[0013] A2. Mix the cyanobacterial raw material powder with deionized water at a ratio of 1:20 g / mL, add 0.5-1 wt% of mixed enzyme, and enzymatically hydrolyze for 1-2 hours at 40±2℃ and pH 4.0±0.5. After the enzymatic hydrolysis is completed, heat to inactivate the enzyme, homogenize under high pressure 2-3 times to obtain cyanobacterial slurry, filter out insoluble matter, freeze dry to obtain cyanobacterial extract.

[0014] Preferably, the mixed enzymes include cellulase, pectinase, and acidic protease in a mass ratio of 1:1-3:0.5-1.5.

[0015] Preferably, the high-pressure homogenization pressure is 60-80 MPa, the temperature is 30-40℃, and the homogenization time for a single cycle is 10-20 min.

[0016] Secondly, the present invention also provides the use of the composition containing blue copper peptides for repairing photothermal damage described in the first aspect in the preparation of skin care products.

[0017] Preferably, the skincare product is a leave-on skincare product.

[0018] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0019] This invention constructs a multi-dimensional repair system for regulating the expression of heat shock proteins (HSPs) through the synergistic combination of a complex peptide, galactosomal yeast fermentation product filtrate, and cyanobacterial extract. The complex peptide, specifically the copper peptide, acts as a signaling molecule, targeting and binding to heat shock factor-1 (HSF-1) to initiate gene transcription. The yeast peptide enhances HSF-1 stability by phosphorylating key proteins in the MAPK / ERK pathway. Hexapeptide-11 inhibits the ubiquitin-proteasome system to prolong the half-life of HSPs. These three components form a cascade effect of "activation-enhancement-retention," working together to... It promotes the expression of HSFs; on this basis, galactosomal yeast filtrate reduces intracellular ROS levels by providing antioxidant components, alleviating the inhibition of HSF-1 by oxidative stress. Its small molecule active substances and polysaccharide components in cyanobacteria extract synergistically promote transdermal absorption, while cyanobacteria extract promotes post-translational modification of HSFs through the TLR4 / NF-κB pathway, ultimately achieving a dual protection mechanism of "internal repair of HSFs - external physical barrier reinforcement of stratum corneum hydration", forming a whole-chain regulation from signal molecule stress response to tissue barrier reconstruction. Detailed Implementation

[0020] The following embodiments are only used to illustrate the technical solutions of the present invention more clearly, and are therefore only examples and should not be used to limit the scope of protection of the present invention.

[0021] Those skilled in the art will understand that the present invention can be practiced even without certain specific details. In other embodiments, methods, means, apparatus, and steps well-known to those skilled in the art have not been described in detail to highlight the spirit of the invention. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art. Unless otherwise specified, all units used in this specification are International Standard Units (SI), and all numerical values ​​and ranges appearing in this invention should be understood to include systematic errors unavoidable in industrial production.

[0022] Unless otherwise specified, the experimental methods used in the following examples are all conventional methods; the materials, reagents, or instruments used, unless otherwise specified by the manufacturer, are all commercially available; and the conditions not specified in the examples are all performed under conventional conditions or conditions recommended by the manufacturer. Furthermore, this invention does not limit the source of the raw materials used; unless otherwise specified, the raw materials used in this invention are all commercially available products commonly found in this technical field. Unless otherwise specified, the "ratio" referred to in the following examples refers to a ratio of parts by mass.

[0023] The raw materials used in this invention and their sources are as follows:

[0024] Blue copper peptide: purchased from Shenzhen Ruidelin Biotechnology Co., Ltd.

[0025] Yeast polypeptide: purchased from Huibai Biotechnology (Guangzhou) Co., Ltd.

[0026] Hexapeptide-11: Purchased from Shenzhen Ruidelin Biotechnology Co., Ltd.

[0027] Galactosomal yeast fermentation product filtrate: purchased from Huibai Biotechnology (Guangzhou) Co., Ltd.

[0028] Suizenji Blue-Green Algae Powder: Purchased from Guangzhou Huayang Biotechnology Co., Ltd.

[0029] Spirulina platensis powder: purchased from Gansu Yishengxiang Biotechnology Co., Ltd.

[0030] Black moss powder: purchased from Ningxia Xiangcao Biotechnology Co., Ltd.

[0031] Cellulase: Purchased from Xiasheng Enzyme, model: FDG-2225, enzyme activity: 11000U / g.

[0032] Pectinase: Purchased from Xiasheng Enzyme, model: FDG-2259, enzyme activity: 300000U / g.

[0033] Acidic protease: purchased from Xiasheng Enzyme, model: FDG-2237, enzyme activity: 500000U / g.

[0034] Before use, the cellulase, pectinase, and acidic protease mentioned above were diluted with ultrapure water to an enzyme activity of 10,000 U / g.

[0035] All other raw materials and reagents required for the experiment were commercially available products.

[0036] Preparation of cyanobacteria extract

[0037] Blue algae extract 1

[0038] Its preparation method includes the following steps:

[0039] A1. Mix Suizenji blue algae powder, Spirulina platensis powder, and Nostoc commune powder in a mass ratio of 1:1:4 to obtain blue algae raw material powder;

[0040] A2. Mix the cyanobacterial raw material powder with deionized water at a ratio of 1:20 g / mL, add 0.8 wt% mixed enzyme, and enzymatically hydrolyze for 1.5 h at 40 ℃ and pH 4.0. After the enzymatic hydrolysis is completed, heat up to inactivate the enzyme, homogenize under high pressure 3 times to obtain cyanobacterial slurry, filter out insoluble matter, freeze dry to obtain cyanobacterial extract 1. The mixed enzyme consists of cellulase, pectinase and acidic protease in a mass ratio of 1:2:1. The pressure of high pressure homogenization is 80 MPa, the temperature is 35 ℃, and the homogenization time is 10 min per cycle.

[0041] Blue algae extract 2

[0042] Its preparation method includes the following steps:

[0043] A1. Mix Suizenji blue algae powder, Spirulina platensis powder, and Nostoc commune powder in a mass ratio of 1:1.5:5 to obtain blue algae raw material powder;

[0044] A2. Mix the cyanobacterial raw material powder with deionized water at a ratio of 1:20 g / mL, add 1 wt% mixed enzyme, and enzymatically hydrolyze for 2 hours at 42℃ and pH 3.5. After enzymatic hydrolysis, heat to inactivate the enzyme, homogenize twice under high pressure to obtain cyanobacterial slurry, filter out insoluble matter, freeze dry to obtain cyanobacterial extract 2. The mixed enzyme consists of cellulase, pectinase and acidic protease in a mass ratio of 1:3:1.5. The pressure of high pressure homogenization is 70 MPa, the temperature is 40℃, and the single homogenization time is 15 min.

[0045] Blue algae extract 3

[0046] Its preparation method includes the following steps:

[0047] A1. Mix Suizenji blue algae powder, Spirulina platensis powder, and Nostoc commune powder in a mass ratio of 1:0.5:3 to obtain blue algae raw material powder;

[0048] A2. Mix the cyanobacterial raw material powder with deionized water at a ratio of 1:20 g / mL, add 0.5 wt% mixed enzyme, and enzymatically hydrolyze for 1 h at 38℃ and pH 4.5. After the enzymatic hydrolysis is completed, raise the temperature to inactivate the enzyme, homogenize twice under high pressure to obtain cyanobacterial slurry, filter out insoluble matter, freeze dry to obtain cyanobacterial extract 3. The mixed enzyme consists of cellulase, pectinase and acidic protease in a mass ratio of 1:1:0.5. The pressure of high pressure homogenization is 60 MPa, the temperature is 30℃, and the single homogenization time is 20 min.

[0049] Blue-green algae extract ①: Unlike blue-green algae extract 1, the blue-green algae raw material powder is made by mixing Spirulina platensis powder and Nostoc moss powder in a mass ratio of 1:4. The remaining steps are the same as those for blue-green algae extract 1.

[0050] Blue-green algae extract ②: Unlike blue-green algae extract 1, the blue-green algae raw material powder is made by mixing Suizenji blue-green algae powder and nori powder in a mass ratio of 1:4. The remaining steps are the same as those for blue-green algae extract 1.

[0051] Blue-green algae extract ③: Unlike blue-green algae extract 1, the blue-green algae raw material powder is made by mixing Suizenji blue-green algae powder and Spirulina platensis powder in a mass ratio of 1:1. The remaining steps are the same as those for blue-green algae extract 1.

[0052] Blue-green algae extract ④: Unlike blue-green algae extract 1, the blue-green algae raw material powder is made by mixing Suizenji blue-green algae powder, Spirulina platensis powder and Nostoc commune powder in a mass ratio of 1:4:1. The remaining steps are the same as those for blue-green algae extract 1.

[0053] Preparation of a composition containing copper peptides for repairing photothermal damage

[0054] Composition 1: Composed of a complex polypeptide, galactosomal yeast fermentation product filtrate, and cyanobacterial extract 1 in a mass ratio of 1:7:5, wherein the complex polypeptide is composed of blue copper peptide, yeast polypeptide, and hexapeptide-11 in a mass ratio of 1:0.4:0.6.

[0055] Preparation method: Mix the above components evenly according to the formula amount, and store in a sealed container away from light.

[0056] Composition 2: Composed of a complex polypeptide, galactosomal yeast fermentation product filtrate, and cyanobacterial extract 2 in a mass ratio of 1:6:4, wherein the complex polypeptide is composed of blue copper peptide, yeast polypeptide, and hexapeptide-11 in a mass ratio of 1:0.3:0.5, and the preparation method is the same as that of composition 1.

[0057] Composition 3: Composed of a complex polypeptide, galactosomal yeast fermentation product filtrate, and cyanobacterial extract 3 in a mass ratio of 1:8:6, wherein the complex polypeptide is composed of blue copper peptide, yeast polypeptide, and hexapeptide-11 in a mass ratio of 1:0.5:0.7, and the preparation method is the same as that of composition 1.

[0058] Composition ①: Unlike composition 1, cyanobacterial extract ① is used to replace cyanobacterial extract 1 in an equal amount, while the remaining components and preparation methods are the same as those of composition 1.

[0059] Composition ②: Unlike composition 1, cyanobacterial extract ② is used to replace cyanobacterial extract 1 in an equal amount, while the remaining components and preparation methods are the same as those of composition 1.

[0060] Composition ③: Unlike composition 1, cyanobacterial extract ③ is used to replace cyanobacterial extract 1 in an equal amount, while the remaining components and preparation methods are the same as those of composition 1.

[0061] Composition ④: Unlike Composition 1, cyanobacterial extract ④ is used to replace cyanobacterial extract 1 in an equal amount, while the remaining components and preparation methods are the same as those of Composition 1.

[0062] Composition A: Unlike Composition 1, Composition A does not contain complex polypeptides, but is composed of galactosomal yeast fermentation product filtrate and cyanobacterial extract 1 in a mass ratio of 7:5; the preparation method is the same as that of Composition 1.

[0063] Composition B: Unlike Composition 1, Composition B does not contain galactosomal yeast fermentation product filtrate, but is composed of a complex polypeptide and cyanobacterial extract 1 in a mass ratio of 1:5. The complex polypeptide is composed of blue copper peptide, yeast polypeptide and hexapeptide-11 in a mass ratio of 1:0.4:0.6. The preparation method is the same as that of Composition 1.

[0064] Composition C differs from Composition 1 in that it does not contain cyanobacterial extract 1, but is composed of a complex polypeptide and galactosomal yeast fermentation product filtrate in a mass ratio of 1:7. The complex polypeptide is composed of blue copper peptide, yeast polypeptide, and hexapeptide-11 in a mass ratio of 1:0.4:0.6. The preparation method is the same as that of Composition 1.

[0065] Composition D differs from Composition 1 in that it consists of a complex polypeptide, galactosomal yeast fermentation product filtrate, and cyanobacterial extract 1 in a mass ratio of 1:5:7. The complex polypeptide consists of blue copper peptide, yeast polypeptide, and hexapeptide-11 in a mass ratio of 1:0.4:0.6. The preparation method is the same as that of Composition 1.

[0066] Composition E: Unlike Composition 1, the complex polypeptide of Composition E does not contain blue copper peptide, but is composed of yeast polypeptide and hexapeptide-11 in a mass ratio of 0.4:0.6; the preparation method is the same as that of Composition 1.

[0067] Composition F differs from Composition 1 in that the complex polypeptide in Composition F does not contain yeast polypeptide, but is composed of blue copper peptide and hexapeptide-11 in a mass ratio of 1:0.6; the preparation method is the same as that of Composition 1.

[0068] Composition G differs from Composition 1 in that the complex polypeptide in Composition G does not contain hexapeptide-11, but is composed of blue copper peptide and yeast polypeptide in a mass ratio of 1:0.4; the preparation method is the same as that of Composition 1.

[0069] Composition H: The complex polypeptide of composition H is composed of blue copper peptide, yeast polypeptide and hexapeptide-11 in a mass ratio of 1:0.6:0.4. The other components and preparation methods are the same as those of composition 1.

[0070] Experimental Example 1

[0071] Heat shock proteins are a class of proteins synthesized in large quantities under cellular stress and are widely present in various organisms. When the skin is exposed to external stimuli such as high temperature, infection, ultraviolet radiation, or chemical stimulation, epidermal cells rapidly initiate the synthesis mechanism of these proteins to help repair damaged proteins, maintain cellular homeostasis, and maintain the skin barrier. Therefore, increasing the secretion of HaCaT heat shock proteins helps to combat protein denaturation caused by ultraviolet / heat, promote the repair and clearance of damaged proteins, and thus promote the self-repair of HaCaT cells.

[0072] Reagents: 0.05% (EDTA-containing) trypsin was manufactured by Thermo Fisher Scientific (China) Co., Ltd., catalog number: 25300120; DMEM medium was manufactured by Thermo Fisher Scientific (China) Co., Ltd., catalog number: 12430054; penicillin antibody was manufactured by Thermo Fisher Scientific (China) Co., Ltd., catalog number: 15140148; CCK-8 kit was manufactured by Beyotime Biotechnology Co., Ltd., catalog number: C0037; fetal bovine serum was manufactured by Beyotime Biotechnology Co., Ltd., catalog number: C0226S; phosphate buffer was manufactured by Beyotime Biotechnology Co., Ltd., catalog number: C0221A; cell lysis buffer was manufactured by Beyotime Biotechnology Co., Ltd., catalog number: P0037-100ml.

[0073] Test sample solution: Dilute compositions 1-3, 1-4, and AH with DMSO to a mass fraction of 10%, and then prepare a test sample solution with a volume fraction of 0.05% using serum-free DMEM medium. Filter to sterilize and set aside for use.

[0074] Test Method: HaCaT cells were cultured in DMEM medium containing 10% v / v fetal bovine serum and 1% v / v antibiotics. Cells were grown in an incubator at 37°C and 5% CO2 saturated humidity. When cell confluence reached 85% or higher, the logarithmic growth phase cells were digested with 0.05% trypsin, and the digestion reaction was terminated with DMEM medium containing 10% v / v fetal bovine serum. Cells were resuspended in serum-free DMEM medium and the cell concentration was adjusted to 2 × 10⁶ cells / mL. 5 Cells were seeded at a density of 2 mL / mL in each well of a six-well plate. Two control and sample groups were established. The sample group received 200 μL of the test solution, while the control group received an equal volume of serum-free DMEM medium. Each control and sample group was in triplicate. Both groups were incubated at 37°C with 5% CO2 for 48 h. The medium was then discarded, and the cells were washed three times with phosphate buffer. Cell lysis buffer was added to each well, and the cells were incubated on ice for 30 min. Cells were gently pipetted to ensure complete lysis, and the liquid in the wells was collected. The liquid was centrifuged at 12000 rpm for 25 min at 4°C, and the supernatant was collected. The human heat shock protein (HSP) content in the cell supernatant was detected using a human heat shock protein (HSP) ELISA kit (Shanghai Enzyme Linked Laboratory, ml063638). The average upregulation rate of HSP was calculated, and the results are shown in Table 1.

[0075] The formula for calculating the average upregulation rate of human heat shock protein levels is as follows:

[0076] Upregulation rate of human heat shock protein content (%) = (heat shock protein content of sample group - heat shock protein content of blank group) / heat shock protein content of blank group × 100%.

[0077] Average upregulation rate of human heat shock protein = Sum of upregulation rates of human heat shock protein in parallel wells of each group / Number of parallel wells

[0078] Table 1. Effect of the composition on the upregulation rate of heat shock protein content

[0079] Group Average upregulation rate of human heat shock protein content / % Blank group - Composition 1 27.33 Composition 2 24.26 Composition 3 25.12 Composition ① 19.17 Composition ② 18.54 Composition ③ 19.31 Composition ④ 21.92 Composition A 12.22 Composition B 14.37 Composition C 15.04 Composition D 19.23 Composition E 15.18 Composition F 16.82 Composition G 17.36 Composition H 20.62

[0080] As shown in Table 1, the composition can significantly increase the heat shock protein content in HaCaT cells. Comparison of compositions 1-3 with compositions ①-④ and composition AH reveals that the raw material composition of the cyanobacterial extract, the composition of the complex polypeptide, and the composition of the entire composition all affect the heat shock protein content in HaCaT cells. Specifically, comparison of composition 1 with compositions ①-④ shows that the cyanobacterial extract prepared using the cyanobacterial raw materials specified in this invention, along with the complex polypeptide and the galactosomal yeast fermentation product filtrate, exhibits a significant synergistic effect, significantly increasing the heat shock protein content in HaCaT cells. Comparison of composition 1 with composition AD shows that the cyanobacterial extract, galactosomal yeast fermentation product filtrate, and complex polypeptide, when used in a specific dosage ratio, have a significant synergistic effect, significantly increasing the heat shock protein content in HaCaT cells. Comparison of composition 1 with composition EH shows that when the complex polypeptide in the composition is composed of tripeptide-1 copper, yeast polypeptide, and hexapeptide-11 in the specified mass ratio specified in this invention, it significantly increases the heat shock protein content in HaCaT cells.

[0081] Experimental Example 2

[0082] This experimental example aims to quantitatively verify the promoting effect of the composition prepared in this invention on keratinocyte proliferation through cell experiments.

[0083] Keratinocytes are cells that have a decisive influence on the skin barrier. Ultraviolet radiation easily generates free radicals and damages keratinocytes. Therefore, keratinocyte proliferation has a certain indicative effect on the repair of photothermal damage.

[0084] Reagents: 0.05% (EDTA-containing) trypsin was manufactured by Thermo Fisher Scientific (China) Co., Ltd., catalog number: 25300120; DMEM medium was manufactured by Thermo Fisher Scientific (China) Co., Ltd., catalog number: 12430054; penicillin antibody was manufactured by Thermo Fisher Scientific (China) Co., Ltd., catalog number: 15140148; CCK-8 kit was manufactured by Beyotime Biotechnology Co., Ltd., catalog number: C0037; fetal bovine serum was manufactured by Beyotime Biotechnology Co., Ltd., catalog number: C0226S; phosphate buffer was manufactured by Beyotime Biotechnology Co., Ltd., catalog number: C0221A.

[0085] Test sample solution: Dilute compositions 1-3, ①-④, and AH with DMSO to a mass fraction of 10%, then prepare a test sample solution with a volume percentage of 0.05% using serum-free DMEM medium. Filter to sterilize and set aside for use.

[0086] HaCaT cells were cultured in DMEM medium containing 10% v / v fetal bovine serum and 1% v / v antibiotics. Cells were grown in an incubator at 37°C and 5% CO2 saturated humidity. When cell confluence reached 85% or higher, the logarithmic growth phase cells were digested with 0.05% trypsin, and the digestion was terminated with DMEM medium containing 10% v / v fetal bovine serum. The cells were resuspended in serum-free DMEM medium, and the cell suspension concentration was adjusted to 7 × 10⁶ cells / mL. 4 Cells were seeded at a density of 100 μL / well in 96-well plates and incubated at 37°C with 5% CO2 for 12 h. After incubation, the old culture medium was removed, and the cells were washed twice with phosphate-buffered saline (PBSS). Then, 100 μL of PBSS was added. Both the model and experimental groups were irradiated with UVB at a dose of 15 mJ / cm². 2 The negative control group was not irradiated. Phosphate buffer was discarded. Serum-free DMEM medium was added to the model group and negative control group. 100 μL of the filtered and sterilized test sample solution prepared above was added to each well in the experimental group, with six parallel wells for each test sample. The blank control group was cell-free and contained 100 μL of serum-free DMEM medium. The experimental group, model group, negative control group, and blank control group were incubated at 37℃ and 5% CO2 for 24 h. Then, 10 μL of CCK-8 solution was added to each well, and the cells were incubated for another 3 h. The absorbance was measured at 450 nm, and the mean cell proliferation rate of each group was calculated. The results are shown in Table 2.

[0087] The formula for calculating the mean cell proliferation rate is as follows:

[0088] Cell proliferation rate (%) = (A experimental group or model group - A blank control group) / (A negative control group - A blank control group) × 100%.

[0089] In the formula: A_experimental group or model group is the absorbance value of the experimental group or model group; A_blank control group is the absorbance value of the blank control group; A_negative control group is the absorbance value of the negative control group.

[0090] Mean cell proliferation rate = Sum of cell proliferation rates in parallel wells of each group / Number of parallel wells in each group

[0091] Table 2. Effect of the composition on HaCaT proliferation rate under UVB irradiation.

[0092] Group Mean cell proliferation rate (%) Model group 42.57 Composition 1 83.44 Composition 2 81.46 Composition 3 80.37 Composition ① 72.24 Composition ② 69.31 Composition ③ 74.56 Composition ④ 77.39 Composition A 49.71 Composition B 52.33 Composition C 56.84 Composition D 73.29 Composition E 57.51 Composition F 62.42 Composition G 64.59 Composition H 75.34

[0093] As shown in Table 2, the composition can significantly increase the proliferation rate of HaCaT cells after UVB exposure. Comparison of compositions 1-3 with compositions ①-④ and AH reveals that the raw material composition of the cyanobacterial extract, the composition of the complex polypeptide, and the composition of the entire composition all affect the proliferation rate of HaCaT cells after UVB exposure. Specifically, comparison of composition 1 with compositions ①-④ shows that the cyanobacterial extract prepared using the cyanobacterial raw materials specified in this invention, along with the complex polypeptide and the galactosomal yeast fermentation product filtrate, exhibits a significant synergistic effect, significantly increasing the proliferation rate of HaCaT cells after UVB exposure. Comparison of composition 1 with composition AD shows that the cyanobacterial extract, galactosomal yeast fermentation product filtrate, and complex polypeptide, when used in a specific dosage ratio, have a significant synergistic effect, significantly increasing the proliferation rate of HaCaT cells after UVB exposure. Comparison of composition 1 with composition EH shows that when the complex polypeptide in the composition is composed of copper peptide, yeast polypeptide, and hexapeptide-11 in the specified mass ratio specified in this invention, it significantly increases the proliferation rate of HaCaT cells after UVB exposure.

[0094] Experimental Example 3

[0095] This experimental example aims to quantitatively verify the safety (Experiment A) and barrier repair efficacy (Experiment B) of the composition prepared in this invention in actual use through subjective human evaluation, as detailed below:

[0096] Experiment A:

[0097] The irritation of cosmetics was evaluated using the 2015 Cosmetic Safety Technical Specifications as a reference standard. The test method was a skin patch test, and the test was conducted on volunteers aged 16-65 years who were randomly distributed. The volunteers were divided into groups according to the number of test substances, with each group corresponding to 1 test substance and 10 people in each group.

[0098] Test substances: Dilute compositions 1-3, compositions ①-④, and composition AH with deionized water to a mass fraction of 5% and prepare as needed.

[0099] Test method: Place the test substance into a patch applicator (0.025g). Cover the patch applicator with non-irritating adhesive tape and apply it to the flexor side of the subject's forearm. Gently press with the palm to ensure even adhesion to the skin surface. Leave on for 24 hours. Remove the patch applicator and observe the skin reaction 30 minutes after the pressure mark disappears. If the result is negative, observe again at 24 and 48 hours after the patch test.

[0100] Evaluation criteria:

[0101] Grade 0: Negative reaction;

[0102] Grade 1: Suspicious reaction, with only slight erythema;

[0103] Grade 2: Weak positive reaction, erythema, infiltration, edema, and possible papules;

[0104] Grade 3: Strong positive reaction, erythema, infiltration, edema, papules may be present, and the reaction may extend beyond the test area;

[0105] Grade 4: Extremely positive reaction, with obvious erythema, severe infiltration, edema, confluent herpes, and reaction extending beyond the test area.

[0106] Test results: All subjects had negative skin reactions.

[0107] Experiment B:

[0108] Test samples: Dilute compositions 1-3, compositions ①-④, and composition AH with deionized water to a mass fraction of 3% and prepare as needed.

[0109] Experimental Methods: Each experimental group had one test sample, with 10 volunteers in each group. A 5cm x 5cm area was marked on the same location on the back of each volunteer as the test area. The volunteers were then exposed to sunlight until a noticeable burning sensation and redness appeared on the test area before the test sample was applied. The test area was applied twice daily for 15 consecutive days (no further sun exposure was allowed during the test sample application period). Transdermal water loss (TEWL) was measured after sun exposure and 15 days after application. The average TEWL value reduction rate was calculated using the following formula. The test results are shown in Table 3 below.

[0110] TEWL decrease rate = (D0 - D15) / D0 × 100%

[0111] Where D0 is the TEWL value of the skin in the test area after sun exposure; D15 is the TEWL value of the skin in the test area after 15 days of use.

[0112] Average TEWL decline rate = Sum of TEWL decline rates of all groups / Number of volunteers in each group

[0113] Table 3 Human efficacy test of the composition

[0114] Test sample Number of volunteers / person TEWL average decline rate / % Composition 1 10 40.26 Composition 2 10 39.38 Composition 3 10 39.52 Composition ① 10 30.19 Composition ② 10 29.72 Composition ③ 10 32.33 Composition ④ 10 33.93 Composition A 10 17.39 Composition B 10 19.22 Composition C 10 20.87 Composition D 10 31.46 Composition E 10 22.26 Composition F 10 25.41 Composition G 10 28.53 Composition H 10 32.91

[0115] As shown in Table 3, compositions 1-3, 1-4, and AH all have the effect of reducing the TEWL value of sunburned skin. Comparing the results of composition 1 with those of compositions 1-4 and AH, it can be seen that the raw materials defined in this invention have a significant synergistic effect, and their combined use can significantly reduce transepidermal water loss.

[0116] The above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit it. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the spirit and scope of the present invention, and all such modifications and substitutions should be covered within the scope of the claims of the present invention. Technical aspects, shapes, and structures not described in detail in this invention are all well-known technologies.

Claims

1. A composition containing a copper peptide for repairing photothermal damage, characterized in that, The composition comprises a complex polypeptide, galactosomal yeast fermentation product filtrate, and cyanobacterial extract. The complex polypeptide comprises copper peptide, yeast polypeptide, and hexapeptide-11 in a mass ratio of 1:0.3-0.5:0.5-0.

7. The mass ratio of the complex polypeptide, galactosomal yeast fermentation product filtrate, and cyanobacterial extract is 1:6-8:4-6. The cyanobacterial raw materials in the cyanobacterial extract include Suizenji cyanobacterial powder, Spirulina platensis powder, and Nostoc moss powder in a mass ratio of 1:0.5-1.5:3-5.

2. The composition according to claim 1, characterized in that, The preparation method of the cyanobacteria extract includes the following steps: A1. Mix Suizenji blue algae powder, Spirulina platensis powder, and Nostoc commune powder evenly according to the formula to obtain blue algae raw material powder; A2. Mix the cyanobacterial raw material powder with deionized water at a ratio of 1:20 g / mL, add 0.5-1 wt% of mixed enzyme, and enzymatically hydrolyze for 1-2 hours at 40±2℃ and pH 4.0±0.

5. After the enzymatic hydrolysis is completed, heat to inactivate the enzyme, homogenize under high pressure 2-3 times to obtain cyanobacterial slurry, filter out insoluble matter, freeze dry to obtain cyanobacterial extract.

3. The composition according to claim 2, characterized in that, The mixed enzymes include cellulase, pectinase, and acidic protease in a mass ratio of 1:1-3:0.5-1.

5.

4. The composition according to claim 2, characterized in that, The high-pressure homogenization is carried out at a pressure of 60-80 MPa, a temperature of 30-40℃, and a single homogenization time of 10-20 min.

5. The use of the composition according to any one of claims 1-4 in the preparation of skin care products.

6. The application as described in claim 5, characterized in that, The skincare product in question is a leave-on skincare product.