Multi-component composite composition with energy protection function and application thereof

Abstract

The invention discloses a multi-component composite composition with an energy protection function and application of the multi-component composite composition. The multi-component composite composition comprises at least two of the following components: saccharomycetes polypeptide, a polyamino acid polysaccharide condensation compound and galactose yeast-like bacterium fermentation product filtrate. The components in the multi-component composite composition have a synergistic interaction effect, so that oxidation resistance can be generated at the same time; mitochondria is protected; anti-stress performance is realized; light damage or light aging resistance; anti-inflammatory or inflammatory injury; promoting skin barrier generation or enhancing skin barrier defense; therefore, the use requirements of various types of consumer groups are met.

Description

Technical Field

[0001] The present invention relates to the technical field of cosmetics, and particularly relates to a multi-component composite composition with an energy protection function and its application. Background Art

[0002] With the upgrading of consumers' demands for the efficacy and safety of cosmetics, bio-fermented active ingredients have gradually become a hot spot in the development of anti-aging and repair products due to their natural sources, high biocompatibility, and multiple skin care effects.

[0003] In related technologies, yeast fermentation products and their derivatives have been widely used in the fields of moisturizing, barrier repair, and antioxidant. For example, patent CN20XX123456A discloses the application of a single yeast fermentation filtrate in regulating skin microecology. However, traditional fermentation components often face technical bottlenecks such as low transdermal absorption rate due to the too large molecular weight of active substances (such as polysaccharide substances), single functional dimension (such as only focusing on moisturizing or antioxidant), and instability of activity when multi-components are compounded. Especially when active substances such as polypeptides, polysaccharides, and amino acids coexist, it is easy to cause system flocculation or efficacy antagonism due to intermolecular forces, which significantly restricts the synergistic potential of multi-component composite products.

[0004] Based on this, developing a cosmetic composition with a multi-component composite system that can effectively overcome the above problems, achieve the stable coexistence of multiple active ingredients, and can synergistically improve the skin barrier reconstruction and the repair efficiency of photo-damage, and has anti-stress and anti-irritation effects, has become a technical difficulty that needs to be urgently broken through in this field. Summary of the Invention

[0005] The present invention aims to at least solve one of the above technical problems existing in the prior art. For this reason, the purpose of the present invention is to provide a multi-component composite composition with an energy protection function and its application. The multi-component composite composition contains three different types of components: yeast polypeptides, polyamino acid polysaccharide condensates, and filtrate of Saccharomyces cerevisiae fermentate. Through the synergistic effect among the three types of components, the improvement of skin barrier reconstruction and photo-damage repair efficiency is achieved, and it has significant anti-stress and anti-irritation effects, which can meet the usage requirements of various types of consumer groups.

[0006] In the first aspect of the present invention, a composition is provided, and the composition includes at least two of the following components:

[0007] Yeast polypeptides, polyamino acid polysaccharide condensates, and filtrate of Saccharomyces cerevisiae fermentate.

[0008] In some embodiments of the present invention, the composition includes a combination of yeast polypeptides, polyamino acid polysaccharide condensates, and filtrate of Saccharomyces cerevisiae fermentate.

[0009] In some embodiments of the present invention, the composition comprises a combination of yeast polypeptides and a filtrate of the fermentation product of Saccharomycopsis.

[0010] In some embodiments of the present invention, in the composition, the mass ratio of yeast polypeptides to the filtrate of the fermentation product of Saccharomycopsis is 0.00001 - 0.00010:1.

[0011] In some embodiments of the present invention, in the composition, the mass ratio of yeast polypeptides to the filtrate of the fermentation product of Saccharomycopsis is 0.00005 - 0.00010:1.

[0012] In some embodiments of the present invention, in the composition, the mass ratio of yeast polypeptides to the filtrate of the fermentation product of Saccharomycopsis is 0.00005:1.

[0013] In some embodiments of the present invention, in the composition, the mass ratio of yeast polypeptides, polyamino acid polysaccharide condensate, and the filtrate of the fermentation product of Saccharomycopsis is 0.00001 - 0.00010:0.0001 - 0.0002:1.

[0014] In some embodiments of the present invention, in the composition, the mass ratio of yeast polypeptides, polyamino acid polysaccharide condensate, and the filtrate of the fermentation product of Saccharomycopsis is 0.00005 - 0.00010:0.00015 - 0.0002:1.

[0015] In some embodiments of the present invention, in the composition, the mass ratio of yeast polypeptides, polyamino acid polysaccharide condensate, and the filtrate of the fermentation product of Saccharomycopsis is 0.00005:0.00015:1.

[0016] In some embodiments of the present invention, the composition comprises:

[0017] 0.0000005 - 0.00005 parts by mass of yeast polypeptides, 0.000005 - 0.00005 parts by mass of polyamino acid polysaccharide condensate, and 0.01 - 0.5 parts by mass of the filtrate of the fermentation product of Saccharomycopsis.

[0018] In some embodiments of the present invention, the composition comprises:

[0019] 0.0000025 parts by mass of yeast polypeptides (SP), 0.0000075 parts by mass of polyamino acid polysaccharide condensate (SC), and 0.05 parts by mass of the filtrate of the fermentation product of Saccharomycopsis (PI).

[0020] In some embodiments of the present invention, the composition comprises:

[0021] By mass, 0.00001 part of yeast polypeptide (SP), 0.00003 part of polyamino acid polysaccharide condensate (SC), and 0.2 part of filtrate of Saccharomycopsis ferment lysate (PI).

[0022] In some embodiments of the present invention, the composition comprises:

[0023] By mass, 0.0000005 - 0.00005 part of yeast polypeptide and 0.01 - 0.5 part of filtrate of Saccharomycopsis ferment lysate.

[0024] In some embodiments of the present invention, the composition comprises:

[0025] By mass, 0.00005 part of yeast polypeptide (SP) and 0.05 part of filtrate of Saccharomycopsis ferment lysate (PI).

[0026] In some embodiments of the present invention, the composition comprises:

[0027] By mass, 0.00001 part of yeast polypeptide (SP) and 0.2 part of filtrate of Saccharomycopsis ferment lysate (PI).

[0028] In the second aspect of the present invention, there is provided a method for preparing the composition described in the above aspect, comprising the following steps:

[0029] Mix yeast polypeptide, filtrate of Saccharomycopsis ferment lysate, and optionally polyamino acid polysaccharide condensate according to the mass ratio described in the above aspect, and you will get it.

[0030] In the present invention, the addition of the polyamino acid polysaccharide condensate is determined based on the selection of the composition. When the prepared composition does not contain polyamino acid polysaccharide condensate, there is no need to add polyamino acid polysaccharide condensate.

[0031] In the third aspect of the present invention, there is provided a cosmetic, which contains the composition described in the above aspect.

[0032] In some embodiments of the present invention, in the cosmetic, by the total mass of the cosmetic, the proportion of the composition is 0.1 - 5%.

[0033] In some embodiments of the present invention, in the cosmetic, by the total mass of the cosmetic, the proportion of the composition is 0.5 - 5%.

[0034] In the fourth aspect of the present invention, there is provided the use of the composition described in the above aspect in the preparation of cosmetics or drugs.

[0035] In some embodiments of the present invention, the cosmetic or drug further contains cosmetically acceptable excipients or pharmaceutically acceptable excipients.

[0036] In some embodiments of the present invention, the cosmetically acceptable excipients include, but are not limited to: preservatives such as phenoxyethanol and parabens; antioxidants such as vitamin E and butylated hydroxytoluene; chelating agents such as sodium ethylenediaminetetraacetate; fragrances, colorants / pigments, pearlescent agents; surfactants such as sodium lauryl sulfate and cocamidopropyl betaine; humectants such as glycerol and hyaluronic acid; pH regulators such as citric acid and sodium lactate.

[0037] In some embodiments of the present invention, the pharmaceutically acceptable excipients include, but are not limited to: antiseptic and antibacterial excipients such as methylparaben, benzalkonium chloride, and phenethyl alcohol; solvents and dispersion media such as water, ethanol / propylene glycol; matrices and carrier materials such as mineral oil / petroleum jelly, polysiloxane (silicone oil), talc / titanium dioxide; stabilizers and antioxidants such as vitamin E / butylated hydroxytoluene, sodium ethylenediaminetetraacetate (EDTA); lubricants and fillers such as magnesium stearate, lactose / microcrystalline cellulose; pH regulators and buffers such as citric acid / sodium hydroxide, phosphate buffer solution.

[0038] In some embodiments of the present invention, the cosmetic or drug has at least one of the following functions (1)-(7):

[0039] (1) Antioxidation;

[0040] (2) Protecting mitochondria;

[0041] (3) Anti-stress;

[0042] (4) Anti-photo-damage or photo-aging;

[0043] (5) Anti-inflammatory or anti-inflammatory injury;

[0044] (6) Promoting skin barrier formation or enhancing skin barrier defense;

[0045] (7) Anti-skin irritation.

[0046] In some embodiments of the present invention, the anti-skin irritation includes: anti-skin cold irritation, anti-skin heat irritation, and anti-skin itching irritation.

[0047] In some embodiments of the present invention, the cosmetic or drug is a topical cosmetic or drug.

[0048] In some embodiments of the present invention, the cosmetics include, but are not limited to: liquid types such as lotion / toner, essence / serum, tincture / spirit; semi-solid types such as cream / ointment, gel, paste; solid types such as powder / foundation powder, mask.

[0049] In some embodiments of the present invention, the drug includes but is not limited to: liquid dosage forms, including solutions, lotions (shakes), tinctures / spirits; semi-solid dosage forms, including ointments, creams / gels, pastes; powders and special dosage forms, including powders, gels, sprays

[0050] The beneficial effects of the present invention are:

[0051] The present invention provides a binary or ternary composite composition, which is composed of two or three different types of substances and produces a synergistic effect, and can simultaneously produce antioxidant; protect mitochondria; resist stress; resist photo-damage or photo-aging; anti-inflammatory or inflammatory damage; promote the generation of skin barrier or enhance skin barrier defense; and anti-skin irritation and other multiple effects, thereby meeting the usage needs of various types of consumer groups. Description of the Drawings

[0052] Figure 1 Results of the inhibition rate of mitochondrial superoxide generation in each experimental group.

[0053] Figure 2 Results of the stress resistance ability in each experimental group.

[0054] Figure 3 Results of the photo-damage resistance ability in each experimental group.

[0055] Figure 4 Results of the photo-aging resistance ability in each experimental group.

[0056] Figure 5 Results of the anti-inflammatory ability in each experimental group.

[0057] Figure 6 Results of the anti-inflammatory ability in each experimental group.

[0058] Figure 7 Results of the photo-aging resistance ability in each experimental group.

[0059] Figure 8 Results of the barrier repair and barrier defense ability in each experimental group.

[0060] Figure 9 Results of the barrier repair and barrier defense ability in each experimental group.

[0061] Figure 10 Results of the heat stimulation resistance ability in each experimental group.

[0062] Figure 11 Results of the anti-itching and anti-pain stimulation ability in each experimental group.

[0063] Figure 12 Results of the cold stimulation resistance ability in each experimental group. Detailed implementation mode

[0064] The content of the present invention will be further described in detail through specific embodiments below. The raw materials, reagents or devices used in the examples and comparative examples can be obtained from conventional commercial channels or by existing technical methods without special instructions. Unless otherwise specified, the test or testing methods are conventional methods in the art.

[0065] In the following examples, yeast polypeptides (SP) were purchased from Huibai Biology, and the CAS number of the multi - amino acid polysaccharide condensate (SC) was 120022 - 92 - 6, which was purchased from Huibai Biology.

[0066] The preparation method of the filtrate of galactose yeast - like bacteria fermentation product (PI) is as follows:

[0067] The activated fermentation strain (Geotrichum candidum) was inoculated into a PDA slant medium and cultured statically at 30°C for 48 h. Then, 5 mL of sterile normal saline was added, and the surface colonies were blown or scraped to obtain a spore suspension. The spore suspension was aspirated and added to 200 mL of fermentation medium (50 g of rice and 8 g of soybean were ground and added to 200 mL of water, stirred evenly, then 200 μL of 20,000 U / mL high - temperature amylase was added, heated to about 90°C, and kept warm for 30 min to complete enzymatic hydrolysis. After enzymatic hydrolysis, it was cooled to 60°C, 10 g of seabuckthorn juice was added, the pH was adjusted to 4 with 1 mol / L phosphoric acid, 0.5 g of 100,000 U / g saccharifying enzyme was added, mixed and kept warm for 1 h, and then the pH was adjusted to 7 with 5 mol / L sodium hydroxide. 20 mg of 100,000 U / g papain was added, and water was added to 1000 mL and then sterilized to obtain the fermentation medium), and cultured at 30°C and 200 rpm for 24 h to obtain a fermentation seed liquid.

[0068] The fermentation seed liquid was inoculated into 200 mL of the above - mentioned new fermentation medium at an inoculation amount of 10%, and cultured at 30°C and 200 rpm for 24 h to obtain a fermentation broth. After the fermentation broth was inactivated at 70°C for 15 min, it was filtered successively with gauze, filter cloth, and filter paper, and the filtrate was the filtrate of galactose yeast - like fermentation.

[0069] Alternatively, a commercially available filtrate of galactose yeast - like bacteria fermentation product can be used for substitution, and its composition is the same as that of the filtrate of galactose yeast - like bacteria fermentation product obtained by the above - mentioned preparation method.

[0070] Example 1

[0071] In this example, a multi - component composite cosmetic composition was provided, and its composition was as follows:

[0072] By mass fraction, 0.0000025 parts of yeast polypeptide (SP), 0.0000075 parts of polyamino acid polysaccharide condensate (SC), and 0.05 parts of filtrate of Saccharomycopsis ferment lysate (PI).

[0073] The preparation method is as follows:

[0074] According to the above mass fraction ratio, mix yeast polypeptide, polyamino acid polysaccharide condensate and filtrate of Saccharomycopsis ferment lysate evenly to obtain the multi-component composite cosmetic composition.

[0075] Example 2

[0076] A multi-component composite cosmetic composition is provided in this example, and its composition is as follows:

[0077] By mass fraction, 0.00001 parts of yeast polypeptide (SP), 0.00003 parts of polyamino acid polysaccharide condensate (SC), and 0.2 parts of filtrate of Saccharomycopsis ferment lysate (PI).

[0078] The preparation method is the same as that of Example 1.

[0079] Example 3

[0080] A multi-component composite cosmetic composition is provided in this example, and its composition is as follows:

[0081] By mass fraction, 0.00005 parts of yeast polypeptide (SP) and 0.05 parts of filtrate of Saccharomycopsis ferment lysate (PI).

[0082] The preparation method is as follows:

[0083] According to the above mass fraction ratio, mix yeast polypeptide and filtrate of Saccharomycopsis ferment lysate evenly to obtain the multi-component composite cosmetic composition.

[0084] Example 4

[0085] A multi-component composite cosmetic composition is provided in this example, and its composition is as follows:

[0086] By mass fraction, 0.00001 parts of yeast polypeptide (SP) and 0.2 parts of filtrate of Saccharomycopsis ferment lysate (PI).

[0087] The preparation method is as follows:

[0088] According to the above mass fraction ratio, mix yeast polypeptide and filtrate of Saccharomycopsis ferment lysate evenly to obtain the multi-component composite cosmetic composition.

[0089] Test Example 1

[0090] In this test example, the antioxidant effects of Example 1 and the corresponding single-component substances were tested. The specific experimental steps were as follows:

[0091] After resuscitating human immortalized epidermal cells (HaCaT), they were cultured routinely. When the cell plating rate reached about 60%, the cells were inoculated into 96-well plates and incubated overnight at 37°C and 5% CO2.

[0092] When the cell plating rate of the 96-well plates reached about 60%, grouping and drug administration were carried out. The experimental groups were divided as follows: the plated cells were divided into a normal control group (NC), a model control group (MC), and a sample group. Among them, the normal control group (NC) and the model control group (MC) were not treated with anything and were continued to be cultured with fresh medium. The sample group was respectively added with a medium containing the multi-component composite cosmetic composition in Example 1, the same content of yeast polypeptide, the same content of polyamino acid polysaccharide condensate, or the same content of filtrate of Saccharomyces galactomyces fermentation product, and continued to be incubated for 2 h at 37°C and 5% CO2.

[0093] Among them, the addition amount of each substance was: calculated by the mass of the medium, the proportion of the corresponding substance was:

[0094] SP+SC+PI group (i.e., the example group): 0.0000025% yeast polypeptide, 0.0000075% polyamino acid polysaccharide condensate, and 0.05% filtrate of Saccharomyces galactomyces fermentation product;

[0095] SP group: 0.0000025% yeast polypeptide; SC group: 0.0000075% polyamino acid polysaccharide condensate; PI group: 0.05% filtrate of Saccharomyces galactomyces fermentation product.

[0096] After the incubation ended, the normal control group (NC) was replaced with fresh medium, and the model control group (MC) was replaced with fresh medium containing 450 μmol / L H2O2, and continued to be incubated for 1 h at 37°C and 5% CO2. For the sample group, fresh medium containing 450 μmol / L H2O2 and the corresponding substance was respectively added, and continued to be incubated for 1 h at 37°C and 5% CO2. For example, for the SP+SC+PI group, fresh medium containing 450 μmol / L H2O2, 0.0000025% yeast polypeptide, 0.0000075% polyamino acid polysaccharide condensate, and 0.05% filtrate of Saccharomyces galactomyces fermentation product was added.

[0097] After the incubation was completed, the culture medium was discarded, and the cells were gently rinsed with 200 μL of PBS. Then, the cells were stained with MitoSOX Red mitochondrial superoxide indicator (purchased from Thermo Fisher) according to the instructions (incubated for 30 min). The mitochondrial superoxide fluorescence intensity of each group was detected using a fluorescence microscope, and the inhibition rate of superoxide generation was calculated.

[0098] Among them, the calculation formula for the inhibition rate of superoxide generation is:

[0099]

[0100] The results are shown in Table 1 and Figure 1 as follows.

[0101] Table 1 Results of the inhibition rate of superoxide generation in each group

[0102] Sample Group Test Result SD P-Value Percentage Normal Control (NC) 47.4 3.14 / / Model Control (MC) 71.8 0.72 <![CDATA[0.0082 ## > 51.5％ SP 61.0 3.00 0.0636 -15.0％ SC 63.2 4.73 0.1526 -12.3％ PI 66.1 2.09 0.4346 -7.9％ SP + SC + PI 52.5 8.91 0.0021** -26.9％

[0103] Among them, compared with NC, #p < 0.05, ##p < 0.01, p < 0.001, #p < 0.0001; compared with MC, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001; four biological replicates (N = 4).

[0104] It can be seen that compared with the model control group (MC), the composition in Example 1 (SP + SC + PI group) can significantly inhibit the mitochondrial superoxide level induced by H2O2, and has good mitochondrial protection and antioxidant effects.

[0105] After testing Example 2, it was found that it had an equivalent effect to Example 1.

[0106] Test Example 2

[0107] In this test example, the anti-stress effects of Example 2 and the corresponding single-component substances were tested. The specific experimental steps were as follows:

[0108] After resuscitating human immortalized epidermal cells (HaCaT), they were cultured routinely. When the cell plating rate reached about 60%, the cells were seeded in 6-well plates and incubated overnight at 37 °C and 5% CO2.

[0109] When the cell seeding rate in the 6-well plate reaches about 60%, group administration is carried out. Experimental grouping: The seeded cells are divided into a normal control group (NC), a model control group (MC), and a sample group. Among them, the normal control group (NC) and the model control group (MC) are not subjected to any treatment and are continuously cultured with fresh medium. The sample group is respectively added with a medium containing the multi-component composite cosmetic composition in Example 2, an equal content of yeast polypeptide, an equal content of polyamino acid polysaccharide condensate, or an equal content of filtrate of galactomyces ferment lysate, and continues to incubate at 37 °C and 5% CO2 for 24 h.

[0110] Among them, the addition amount of each substance is: based on the mass of the medium, the proportion of the corresponding substance is:

[0111] SP+SC+PI group (i.e., the example group): 0.00001% yeast polypeptide, 0.00003% polyamino acid polysaccharide condensate, and 0.2% filtrate of galactomyces ferment lysate;

[0112] SP group: 0.00001% yeast polypeptide; SC group: 0.00003% polyamino acid polysaccharide condensate; PI group: 0.2% filtrate of galactomyces ferment lysate.

[0113] After incubation, discard the medium, and gently rinse the cells 1-2 times with D-Hanks balanced salt solution (D-HBSS). Then expose the model control group (MC) and the sample group to ultraviolet light for 50 s (UVB, 80 mJ / cm 2 ). The normal control group (NC) and the model control group (MC) are added with fresh medium and continue to incubate at 37 °C and 5% CO2 for 24 h. For the sample group, fresh medium containing the corresponding substance is respectively added and continues to incubate at 37 °C and 5% CO2 for 24 h.

[0114] After incubation, change the medium again to completely remove D-HBSS. The normal control group (NC) and the model control group (MC) are not subjected to any treatment and continue to be cultured with fresh medium for 24 h. The sample group is respectively added with the above-mentioned medium containing the multi-component composite cosmetic composition in Example 2, an equal content of yeast polypeptide, an equal content of polyamino acid polysaccharide condensate, or an equal content of filtrate of galactomyces ferment lysate, and continues to incubate at 37 °C and 5% CO2 for 24 h.

[0115] After incubation, use a commercially available kit to extract total RNA, reverse transcription, and detect the expression levels of β-actin (internal reference) and FOXO1 (Forkhead box protein O1) based on q-PCR.

[0116] FOXO1 is an important member of the FoxO transcription factor family and plays a central role in cellular stress response, metabolic regulation, antioxidant defense, and lifespan regulation. Under oxidative stress conditions, FOXO1 is activated through pathways such as AMPK and JNK, translocates to the nucleus to initiate the expression of protective genes, and simultaneously inhibits pro-apoptotic signals (such as Bim) to maintain cell survival. Therefore, the anti-stress ability of a subject can be determined by detecting the expression level of FOXO1.

[0117] The results are shown in Table 2 and Figure 2 as follows.

[0118] Table 2 Results of anti-stress ability of each group

[0119] Sample Group Test Result SD P-Value Percentage Normal Control (NC) 1.3 0.09 / / Model Control (MC) 1.0 0.05 0.0080## -23.1％ SP 1.2 0.14 0.1336 20.3％ SC 1.1 0.16 0.7733 8.9％ PI 1.1 0.10 0.7022 9.3％ SP + SC + PI 1.3 0.04 0.0175* 30.6％

[0120] Among them, compared with NC, #p < 0.05, ##p < 0.01, p < 0.001, #p < 0.0001; compared with MC, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001; four biological replicates (N = 4).

[0121] It can be seen that compared with the control group (MC), the relative expression level of the FOXO1 gene is more significantly increased by the composition (SP + SC + PI) than the single components of SP, SC, and PI, indicating that the combination of SP + SC + PI produces a synergistic effect that is not possessed by the single use of SP, SC, and PI.

[0122] After testing Example 1, it was found that it had an effect comparable to that of Example 2.

[0123] Test Example 3

[0124] In this test example, the anti-photoaging effects of Example 2 and the single-component substances with corresponding contents were tested. The specific experimental steps were as follows:

[0125] After resuscitating human immortalized epidermal cells (HaCaT), they were cultured routinely. When the cell plating rate reached about 60%, the cells were seeded in 6-well plates and incubated overnight at 37°C and 5% CO2.

[0126] When the cell seeding rate in the 6-well plate reaches about 60%, group administration is carried out. Experimental grouping: The seeded cells are divided into a normal control group (NC), a model control group (MC), and a sample group. Among them, the normal control group (NC) and the model control group (MC) are not treated with anything and are continuously cultured with fresh medium. The sample group is respectively added with a medium containing the multi-component composite cosmetic composition in Example 2, an equal content of yeast polypeptide, an equal content of polyamino acid polysaccharide condensate, or an equal content of filtrate of Saccharomyces galactomyces fermentation product, and continue to incubate at 37°C and 5% CO2 for 24 h.

[0127] Among them, the addition amount of each substance is: based on the mass of the medium, the proportion of the corresponding substance is:

[0128] SP+SC+PI group (i.e., the example group): 0.00001% yeast polypeptide, 0.00003% polyamino acid polysaccharide condensate, and 0.2% filtrate of Saccharomyces galactomyces fermentation product;

[0129] SP group: 0.00001% yeast polypeptide; SC group: 0.00003% polyamino acid polysaccharide condensate; PI group: 0.2% filtrate of Saccharomyces galactomyces fermentation product.

[0130] After incubation, discard the medium and gently rinse the cells 1-2 times with D-Hanks balanced salt solution (D-HBSS). Then expose the model control group (MC) and the sample group to ultraviolet light for 50 s (UVB, 80 mJ / cm 2 ). The normal control group (NC) and the model control group (MC) are added with fresh medium and continue to incubate at 37°C and 5% CO2 for 24 h. For the sample group, fresh medium containing the corresponding substance is respectively added and continue to incubate at 37°C and 5% CO2 for 24 h. For example, for the SP+SC+PI group, add fresh medium containing 0.00001% yeast polypeptide, 0.00003% polyamino acid polysaccharide condensate, and 0.2% filtrate of Saccharomyces galactomyces fermentation product.

[0131] After incubation, discard the medium and rinse the cells 1-2 times with D-HBSS. Use a commercially available kit to extract total RNA, reverse transcription, and detect the expression levels of β-actin (internal reference) and PGC-1α (peroxisome proliferator-activated receptor γ coactivator 1α) based on q-PCR.

[0132] Existing studies have shown that PGC-1α constitutes a multi-dimensional protection network against photo-damage by integrating mitochondrial function optimization, antioxidant defense, DNA repair, and anti-inflammatory signals. Its expression is precisely regulated by energy status, epigenetics, and hormonal signals, making it a key target for photoaging intervention. Therefore, the photo-damage resistance ability of the test subjects can be determined by detecting the expression level of PGC-1α.

[0133] The results are shown in Table 3 and Figure 3 as follows.

[0134] Table 3 Results of the photo-damage resistance ability of each group

[0135] Sample Group Test Result SD P-Value Percentage Normal Control (NC) 1.5 0.11 / / Model Control (MC) 1.0 0.13 0.0070## -34.2％ SP 1.3 0.09 0.0449* 26.9％ SC 1.1 0.11 0.4905 12.5％ PI 1.1 0.11 0.6311 10.2％ SP + SC + PI 1.4 0.10 0.0018** 44.0％

[0136] Among them, compared with NC, #p < 0.05, ##p < 0.01, p < 0.001, #p < 0.0001; compared with MC, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001; four biological replicates (N = 4).

[0137] It can be seen that compared with the model control group (MC), the composition (SP + SC + PI) can significantly increase the relative expression level of the PGC-1α gene, and the expression level is more significant than that of the single components of SP, SC, and PI, indicating that the combination of SP + SC + PI produces a synergistic effect that is not possessed by the single use of SP, SC, and PI.

[0138] After testing Example 1, it was found that it had an effect equivalent to that of Example 2.

[0139] Test Example 4

[0140] In this test example, the photoaging resistance effects of Example 1 and the single-component substances with corresponding contents were tested. The specific experimental steps were as follows:

[0141] After resuscitating human skin fibroblasts (HFF-1), they were cultured routinely. When the cell plating rate reached about 60%, the cells were inoculated into 6-well plates and incubated overnight at 37°C and 5% CO2.

[0142] When the cell seeding density in the 6-well plate reaches 30 - 50%, group administration is carried out. Experimental grouping: The seeded cells are divided into a normal control group (NC), a model control group (MC), and a sample group. Among them, the normal control group (NC) and the model control group (MC) are not treated with anything and are continued to be cultured with fresh medium (2 mL / well). The sample group is respectively added with a medium containing the multi-component composite cosmetic composition in Example 1, an equal content of yeast polypeptide, an equal content of polyamino acid polysaccharide condensate, or an equal content of filtrate of Saccharomyces galactomyces fermentation product, and is continued to be incubated at 37 °C and 5% CO₂ for 24 h.

[0143] Among them, the addition amount of each substance is the same as that in Test Example 1.

[0144] After incubation, the model control group (MC) and the sample group are exposed to UVA for irradiation, and the irradiation dose is 30 J / cm 2 . After irradiation, it is continued to be incubated at 37 °C and 5% CO₂ for 24 h.

[0145] After incubation, the medium is discarded, and the cells are rinsed 2 times with PBS. 1 mL of AG RNAex Pro RNA extraction reagent is added to each well, and after blowing and lysing the cells, the samples are collected. Total RNA is extracted using a commercially available kit, reverse transcription is carried out, and the expression levels of β-actin (internal reference) and SIRT3 (silent information regulator 3) are detected based on q-PCR.

[0146] SIRT3 is a deacetylase mainly located in mitochondria in the sirtuin family. Existing studies have shown that SIRT3 plays a key role in anti-photoaging (especially ultraviolet-induced skin damage) by regulating mitochondrial energy metabolism, antioxidant defense, and cellular stress response, and is a key molecular target for anti-photoaging. Therefore, the anti-photoaging ability of the test subject can be determined by detecting the expression level of SIRT3.

[0147] The results are shown in Table 4 and Figure 4 as follows.

[0148] Table 4 Results of anti-photoaging ability of each group

[0149] Sample Group Test Result SD P-Value Percentage Normal Control (NC) 1.0 0.09 / / Model Control (MC) 0.5 0.03 0.0062## -48.0％ SP 0.6 0.07 0.0459* 22.9％ SC 0.6 0.02 0.8492 6.3％ PI 0.6 0.02 0.0701 20.4％ SP + SC + PI 0.7 0.07 0.0062** 30.7％

[0150] Among them, compared with NC, #p < 0.05, ##p < 0.01, p < 0.001, #p < 0.0001; compared with MC, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001; four biological replicates (N = 4).

[0151] It can be seen that, compared with the model control group (MC), the composition (SP + SC + PI) can significantly up-regulate the relative expression level of SIRT3, and the expression level is more significant than that of the single components of SP, SC, and PI, indicating that the combination of SP + SC + PI produces a synergistic effect that is not possessed by the single use of SP, SC, and PI.

[0152] After testing Example 2, it was found that it had an effect comparable to that of Example 1.

[0153] Test Example 5

[0154] In this test example, the anti-inflammatory (inflammatory injury) effects of Example 1 and the single-component substances with corresponding contents were tested. The specific experimental steps were as follows:

[0155] After resuscitating mouse mononuclear macrophages (RAW264), they were cultured routinely. When the cell plating rate reached about 60%, the cells were inoculated into 6-well plates and incubated overnight at 37°C and 5% CO2.

[0156] When the cell plating rate of the 6-well plates reached about 50%, grouping and drug administration were carried out. The experimental groups were divided as follows: the plated cells were divided into a normal control group (NC), a model control group (MC), and a sample group. Among them, the normal control group (NC) and the model control group (MC) were not treated with anything and continued to be cultured with fresh medium. The sample group was respectively added with a medium containing the multi-component composite cosmetic composition in Example 1, the same content of yeast polypeptide, the same content of polyamino acid polysaccharide condensate, or the same content of filtrate of Saccharomyces cerevisiae-like fermented products, and continued to be incubated at 37°C and 5% CO2 for 2 h. Among them, the addition amount of each substance was the same as that in Test Example 1.

[0157] After the incubation was completed, the medium was discarded, and the cells were rinsed 2 times with 200 μL of PBS. The normal control group (NC) was added with fresh medium again and continued to be incubated at 37°C and 5% CO2 for 24 h. For the model control group (MC), fresh medium containing LPS (1 μg / mL) was added, and for the sample group, fresh medium containing LPS (1 μg / mL) and the corresponding substance was added, and they were continued to be incubated at 37°C and 5% CO2 for 24 h. For example, for the SP + SC + PI group, fresh medium containing 1 μg / mL LPS, 0.0000025% yeast polypeptide, 0.0000075% polyamino acid polysaccharide condensate, and 0.05% filtrate of Saccharomyces cerevisiae-like fermented products was added.

[0158] After the incubation was completed, the supernatant was collected by centrifugation. The content was detected using an IL-1β ELISA kit (purchased from Thermo Fisher), and the IL-1β inhibition rate was calculated according to the following formula:

[0159]

[0160] IL-1β is one of the most powerful pro-inflammatory cytokines in the human body. Together with IL-1α, it binds to IL-1R1, thereby initiating an inflammatory response through signal transduction. Therefore, the anti-inflammatory ability of the subject can be determined by the inhibition rate of IL-1β.

[0161] The results are shown in Table 5 and Figure 5 as follows.

[0162] Anti-inflammatory ability results of each group in Table 5

[0163] Sample Group Test Result SD P-Value Percentage Normal Control (NC) 11.2 0.36 / / Model Control (MC) 116.8 4.41 0.0005### 942.9％ SP 100.4 11.81 0.1762 -14.0％ SC 108.6 8.94 0.6865 -7.0％ PI 107.5 14.30 0.5907 -8.0％ SP + SC + PI 91.6 3.27 0.0281* -21.6％

[0164] Among them, compared with NC, #p < 0.05, ##p < 0.01, p < 0.001, #p < 0.0001; compared with MC, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001; four biological replicates (N = 4).

[0165] It can be seen that compared with the model control group (MC), the composition (SP + SC + PI) can significantly inhibit IL-1β, thereby producing an anti-inflammatory effect. In addition, its inhibition rate is more significant than that of the single components of SP, SC, and PI, indicating that the combination of SP + SC + PI produces a synergistic effect that is not possessed by the single use of SP, SC, and PI.

[0166] After testing Example 2, it was found that it had an effect comparable to that of Example 1.

[0167] Test Example 6

[0168] In this test example, the anti-inflammatory (inflammatory injury) effects of Example 1 and the corresponding single-component substances were tested. The specific experimental steps were the same as those in Example 5, except that in this test example, a TNF-α ELISA kit (purchased from ThermoFisher) was used to detect the TNF-α content, and the TNF-α inhibition rate was calculated (the calculation method was the same as that in Example 5).

[0169] TNF-α is one of the recognized markers of the inflammatory response. Therefore, the anti-inflammatory ability of the subject can be determined by the inhibition rate of TNF-α.

[0170] The results are shown in Table 6 and Figure 6 as follows.

[0171] Anti-inflammatory ability results of each group in Table 6

[0172] Sample Group Test Result SD P-Value Percentage Normal Control (NC) 11.2 0.36 / / Model Control (MC) 116.8 4.41 0.0005### 942.9％ SP 100.4 11.81 0.1762 -14.0％ SC 108.6 8.94 0.6865 -7.0％ PI 107.5 14.30 0.5907 -8.0％ SP + SC + PI 91.6 3.27 0.0281* -21.6％

[0173] Among them, compared with the NC group, #p < 0.05, ##p < 0.01, p < 0.001, #p < 0.0001; compared with the MC group, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001; four biological replicates (N = 4).

[0174] It can be seen that compared with the model control group (MC), the composition (SP + SC + PI) can significantly inhibit IL-1β, thus producing an anti-inflammatory effect. In addition, its inhibition rate is more significant than that of the single components of SP, SC, and PI, indicating that the combination of SP + SC + PI produces a synergistic effect that is not possessed by the single use of SP, SC, and PI.

[0175] After testing Example 2, it was found that it had an effect comparable to that of Example 1.

[0176] Test Example 7

[0177] In this test example, the anti-photoaging effects of Example 3 and the single-component substances with corresponding contents were tested. The specific experimental steps were as follows:

[0178] After resuscitating HaCaT cells, they were cultured routinely. When the cell confluence reached about 60%, the cells were seeded in 6-well plates and incubated overnight at 37°C and 5% CO2.

[0179] When the cell confluence in the 6-well plates reached about 50%, grouping and drug administration were carried out. The experimental groups were divided as follows: the cells after seeding were divided into a normal control group (NC), a model control group (MC), and a sample group. Among them, the normal control group (NC) and the model control group (MC) were not treated with anything and were continued to be cultured with fresh medium. The sample group was respectively added with a medium containing the multi-component composite cosmetic composition in Example 3, an equal content of yeast polypeptide, or an equal content of filtrate of Saccharomyces galactomyces ferment, and continued to be incubated at 37°C and 5% CO2 for 24 h.

[0180] Among them, the addition amounts of each substance were as follows:

[0181] SP + PI group (i.e., the example group): 0.0000025% yeast polypeptide and 0.05% filtrate of Saccharomyces galactomyces ferment;

[0182] SP group: 0.0000025% yeast polypeptide; PI group: 0.05% filtrate of Saccharomyces galactomyces ferment.

[0183] After the incubation was completed, the medium was discarded, and the cells were rinsed 2 times with D-HBSS. Then the model control group (MC) and the sample group were exposed to ultraviolet light for 50 s (UVB, 80 mJ / cm 2)。The normal control group (NC) and the model control group (MC) were added with fresh medium and incubated continuously for 24 h under the conditions of 37 °C and 5% CO2. For the sample groups, fresh medium containing the corresponding substances was added respectively and incubated continuously for 24 h under the conditions of 37 °C and 5% CO2.

[0184] After incubation, the medium was discarded, and the cells were rinsed twice with PBS. Total RNA was extracted using a commercially available kit, reverse transcribed, and the expression levels of β-actin (internal reference) and Nrf2 (nuclear factor E2-related factor 2) were detected based on q-PCR.

[0185] Previous studies have pointed out that Nrf2 is the core regulatory factor of the cellular antioxidant defense system and can play a key role in resisting ultraviolet (UV)-induced photoaging by activating multiple antioxidant and detoxifying enzyme genes. Therefore, the photoaging resistance ability of the subjects can be determined by the expression level of Nrf2.

[0186] The results are shown in Table 7 and Figure 7 as follows.

[0187] Table 7 Results of photoaging resistance ability of each group

[0188] Sample Group Test Result SD P-Value Percentage Normal Control (NC) 1.3 0.08 / / Model Control (MC) 1.0 0.07 0.0040## -24.6％ SP 1.3 0.12 0.0253* 26.7％ PI 1.2 0.02 0.1747 16.9％ SP + PI 1.4 0.13 0.0038** 37.8％

[0189] Among them, compared with NC, #p < 0.05, ##p < 0.01, p < 0.001, #p < 0.0001; compared with MC, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001; four biological replicates (N = 4).

[0190] It can be seen that compared with the model control group (MC), the composition (SP + PI) can significantly up-regulate the relative expression level of Nrf2, thereby producing a photoaging resistance effect. In addition, the up-regulation amplitude is more significant than that of the single components of SP and PI, indicating that the combination of SP + PI produces a synergistic effect that is not possessed by the single components when used alone.

[0191] Test Example 8

[0192] In this test example, the effects of promoting skin barrier formation and enhancing skin barrier defense of Example 3 and the single-component substances with corresponding contents were tested. The specific experimental steps were as follows:

[0193] After resuscitating HaCaT, it was cultured routinely. When the cell plating rate reached about 60%, the cells were seeded in a 6-well plate and incubated overnight under the conditions of 37 °C and 5% CO2.

[0194] When the cell seeding rate in the 6-well plate reaches about 50%, group administration is carried out. Experimental grouping: The seeded cells are divided into a normal control group (NC) and a sample group. Among them, the normal control group (NC) is not treated with anything and is continuously cultured with fresh medium. The sample group is respectively added with a medium containing the multi-component composite cosmetic composition in Example 3, an equal content of yeast polypeptide, or an equal content of the filtrate of Saccharomyces galactomyces fermentation product, and continues to incubate at 37°C and 5% CO2 for 24 h.

[0195] Among them, the addition amount of each substance is the same as that in Test Example 7.

[0196] After incubation, discard the medium, wash the cells 2 times with PBS, and use a commercially available kit to extract total RNA, reverse transcription, and detect the expression levels of β-actin (internal reference) and ZO-1 (Zonula Occludens-1) based on q-PCR.

[0197] Previous studies have pointed out that ZO-1 is the core scaffold protein of tight junctions (TJ), which is connected to the cytoskeleton by anchoring transmembrane proteins (such as occludin and claudins), and plays a key role in maintaining the integrity of the epidermal barrier, regulating paracellular permeability, and resisting external stimuli. Therefore, ZO-1 is an important target for barrier repair products (such as sensitive skin care and adjuvant treatment of atopic dermatitis), and the barrier repair and barrier defense capabilities of the test subjects can be determined by the expression of ZO-1.

[0198] The results are shown in Table 8 and Figure 8 as follows.

[0199] Table 8 Results of barrier repair and barrier defense capabilities of each group

[0200] Sample Group Test Result SD P-Value Percentage Normal Control (NC) 1.0 0.11 / / SP 1.3 0.04 0.0123* 28.3％ PI 1.2 0.14 0.0594 20.1％ SP + PI 1.4 0.03 0.0029** 36.7％

[0201] Among them, compared with NC, #p < 0.05, ##p < 0.01, p < 0.001, #p < 0.0001; compared with MC, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001; four biological replicates (N = 4).

[0202] It can be seen that compared with NC, the composition (SP + PI) can significantly up-regulate the relative expression level of ZO-1, thereby producing barrier repair and barrier defense effects. In addition, the up-regulation amplitude is more significant than that of the single components of SP and PI, indicating that the SP + PI combination produces a synergistic effect that is not possessed by the single components.

[0203] Test Example 9

[0204] In this test example, the effects of Example 3 and the single-component substances with corresponding contents on promoting skin barrier formation and enhancing skin barrier defense were tested. The specific experimental steps were the same as those in Test Example 7. The difference was that in this test example, a commercially available endothelial tight junction protein Claudin-1 (CLDN1) detection kit was used to detect the relative expression level of CLDN1.

[0205] CLDN1 is an anchoring transmembrane protein and is also one of the currently recognized markers for determining the barrier repair and barrier defense capabilities of a test subject.

[0206] The results are shown in Table 9 and Figure 9 as follows.

[0207] Table 9 Results of barrier repair and barrier defense capabilities of each group

[0208] Sample Group Test Result SD P-Value Percentage Normal Control (NC) 1.0 0.09 / / SP 1.3 0.13 0.0234* 25.2％ PI 1.2 0.07 0.0951 18.4％ SP + PI 1.3 0.07 0.0159* 26.9％

[0209] Among them, compared with NC, #p < 0.05, ##p < 0.01, p < 0.001, #p < 0.0001; compared with MC, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001; four biological replicates (N = 4).

[0210] It can be seen that compared with NC, the composition (SP + PI) can significantly up-regulate the relative expression level of CLDN1, thereby producing barrier repair and barrier defense effects. In addition, the up-regulation amplitude is more significant than that of the single components of SP and PI, indicating that the combination of SP + PI produces a synergistic effect that is not possessed by the single components when used alone.

[0211] Test Example 10

[0212] In this test example, the anti-thermal stimulation effects of Example 3 and the single-component substances with corresponding contents were tested. The specific experimental steps were as follows:

[0213] After resuscitating HaCaT, it was cultured routinely. When the cell plating rate reached about 60%, the cells were seeded in a 6-well plate and incubated overnight at 37°C and 5% CO2.

[0214] When the cell plating rate of the 6-well plate reached about 50%, grouping and drug administration were carried out. The experimental groups were divided as follows: the plated cells were divided into a normal control group (NC), a model control (MC) group, and a sample group. Among them, the normal control group (NC) and the model control (MC) group were not treated with anything and continued to be cultured with fresh medium. The sample group was respectively added with a medium containing the multi-component composite cosmetic composition in Example 3, an equal content of yeast polypeptide, or an equal content of filtrate of Saccharomyces galactomyces ferment, and continued to be incubated at 37°C and 5% CO2 for 24 h.

[0215] Among them, the addition amounts of each substance are the same as those in Test Example 7.

[0216] After the incubation was completed, the culture medium was discarded, and the cells were rinsed twice with D-HBSS. After replacing with a new culture medium, the normal control group (NC) was continuously incubated at 37 °C and 5% CO2 for 24 h. For the model control (MC) group and the sample group, they were first incubated at a high temperature (44 °C) for 30 min, and then the culture medium containing the corresponding substance was replaced (the model control (MC) group was replaced with the same new culture medium as the normal control group (NC)), and then continuously incubated at 37 °C and 5% CO2 for 24 h.

[0217] After the incubation was completed, the culture medium was discarded, the cells were collected, and a commercially available kit was used to extract total RNA, reverse transcribe, and detect the expression levels of β-actin (internal reference) and HSP70 (heat shock protein 70) based on q-PCR.

[0218] Previous studies have pointed out that HSP70 is the core molecular chaperone for cells to respond to heat stress. It plays a key role in cell protection and damage repair triggered by heat stimulation by maintaining protein homeostasis, inhibiting apoptosis, and regulating inflammatory responses. Therefore, the ability of the test subject to resist heat stimulation can be determined by the expression level of HSP70.

[0219] The results are shown in Table 10 and Figure 10 as follows.

[0220] Table 10 Results of the ability of each group to resist heat stimulation

[0221] Sample Group Test Result SD P-Value Percentage Normal Control (NC) 0.7 0.19 / / Model Control (MC) 1.0 0.09 0.0431# 36.1％ SP 0.8 0.09 0.0963 -18.2％ PI 0.9 0.10 0.1520 -16.1％ SP + PI 0.8 0.09 0.0447* -22.3％

[0222] Among them, compared with NC, #p < 0.05, ##p < 0.01, p < 0.001, #p < 0.0001; compared with MC, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001; four biological replicates (N = 4).

[0223] It can be seen that compared with the model control group (MC), the composition (SP + PI) can significantly reduce the relative expression level of HSP70, thereby producing an anti-heat stimulation effect. In addition, the reduction amplitude is more significant than that of the single components of SP and PI, indicating that the combination of SP + PI produces a synergistic effect that is not possessed by the single components when used alone.

[0224] Test Example 11

[0225] In this test example, the effects of Example 3 and the single-component substances with corresponding contents on resisting pruritus stimulation were tested. The specific experimental steps were as follows:

[0226] After resuscitating HaCaT cells, conduct routine culture. When the cell plating rate reaches about 60%, inoculate the cells into a 6-well plate and incubate overnight at 37°C and 5% CO2.

[0227] When the cell plating rate in the 6-well plate reaches about 50%, perform grouped drug administration. Conduct experimental grouping: Divide the plated cells into a normal control group (NC), a model control (MC) group, and a sample group. Among them, the normal control group (NC) and the model control (MC) group are not treated with anything and continue to be cultured with fresh medium. The sample group is respectively added with a medium containing the multi-component composite cosmetic composition in Example 3, an equal content of yeast polypeptide, or an equal content of the filtrate of Saccharomyces cerevisiae-like bacteria fermentation product, and continue to incubate for 24 h at 37°C and 5% CO2.

[0228] Among them, the addition amount of each substance is the same as that in Test Example 7.

[0229] After incubation, discard the medium and rinse the cells 2 times with D-HBSS. After replacing with a new medium, the normal control group (NC) continues to incubate for 24 h at 37°C and 5% CO2. While the model control (MC) group and the sample group are replaced with a medium containing 15 μM capsaicin and the corresponding substance (the medium of the model control (MC) group only includes capsaicin), and then continue to incubate for 24 h at 37°C and 5% CO2.

[0230] After incubation, discard the medium, collect the cells, and use a commercially available kit to extract total RNA, reverse transcribe, and detect the expression levels of β-actin (internal reference) and TRPV1 (transient receptor potential vanilloid subtype 1) based on q-PCR.

[0231] TRPV1 is a non-selective cation channel protein widely distributed in skin sensory nerve endings, keratinocytes and other parts, and is the core molecular receptor for perceiving stimuli such as pain, itching, and burning. Existing studies have pointed out that the overreaction of sensitive skin to external stimuli (such as stinging, burning, itching) is closely related to TRPV1. Therefore, TRPV1 can be used as an index to evaluate the anti-itching and anti-pain stimuli of the test subjects.

[0232] The results are shown in Table 11 and Figure 11 as follows.

[0233] Table 11 Results of the anti-itching and anti-pain stimuli ability of each group

[0234] Sample Group Test Result SD P-Value Percentage Normal Control (NC) 0.7 0.07 / / Model Control (MC) 1.0 0.11 0.0341# 47.8％ SP 0.8 0.08 0.0484* -20.0％ PI 0.9 0.09 0.1416 -15.0％ SP + PI 0.7 0.06 0.0092** -28.0％

[0235] Among them, compared with NC, #p < 0.05, ##p < 0.01, p < 0.001, #p < 0.0001; compared with MC, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001; four biological replicates (N = 4).

[0236] It can be seen that compared with the model control group (MC), the composition (SP + PI) can significantly reduce the relative expression level of TRPV1, thereby producing an anti-itch and anti-pain stimulation effect. In addition, the reduction amplitude is more significant than that of the single components of SP and PI, indicating that the combination of SP + PI produces a synergistic effect that is not possessed by the single components when used alone.

[0237] Test Example 12

[0238] In this test example, the anti-cold stimulation effects of Example 3 and the single-component substances with corresponding contents were tested. The specific experimental steps are as follows:

[0239] After resuscitating HaCaT, it was cultured routinely. When the cell plating rate reached about 60%, the cells were inoculated into 6-well plates and incubated overnight at 37°C and 5% CO2.

[0240] When the cell plating rate of the 6-well plates reached about 60%, grouping and drug administration were carried out. The experimental groups were divided as follows: the plated cells were divided into a normal control group (NC), a model control (MC) group, and a sample group. Among them, the normal control group (NC) and the model control (MC) group were not treated with anything and were continued to be cultured with fresh medium. The sample group was respectively added with a medium containing the multi-component composite cosmetic composition in Example 3, the same content of yeast polypeptide, or the same content of filtrate of Saccharomyces galactomyces ferment, and continued to be incubated at 37°C and 5% CO2 for 24 h.

[0241] Among them, the addition amount of each substance was the same as that in Test Example 7.

[0242] After the incubation was completed, the medium was discarded, and the cells were rinsed 2 times with D-HBSS. After replacing the new medium, the normal control group (NC) was continued to be incubated at 37°C and 5% CO2 for 24 h. While the model control (MC) group and the sample group were first incubated in a low-temperature environment (37°C) for 30 min, and then the medium containing the corresponding substance was replaced (the model control (MC) group was replaced with the same new medium as the normal control group (NC)), and continued to be incubated at 37°C and 5% CO2 for 24 h.

[0243] After the incubation was completed, the culture medium was discarded, and the cells were collected. Total RNA was extracted using a commercially available kit, reverse transcribed, and the expression levels of β-actin (internal reference) and CIRBP (Cold-Inducible RNA-Binding Protein) were detected based on q-PCR.

[0244] CIRBP is a cold-inducible RNA-binding protein that participates in the regulation of cellular response to cold stress and thus is involved in the adaptation and pathological processes of the skin to cold stimuli. Therefore, CIRBP can be used as an indicator to evaluate the cold-stimulus resistance of the test subjects.

[0245] The results are shown in Table 12 and Figure 12 as follows.

[0246] Table 12 Results of cold-stimulus resistance ability of each group

[0247] Sample Group Test Result SD P-Value Percentage Normal Control (NC) 0.6 0.05 / / Model Control (MC) 1.0 0.05 0.0049## 60.0％ SP 0.9 0.04 0.0170* -14.9％ PI 0.9 0.04 0.0342* -13.2％ SP + PI 0.8 0.07 0.0019** -22.1％

[0248] Among them, compared with NC, #p < 0.05, ##p < 0.01, p < 0.001, #p < 0.0001; compared with MC, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001; four biological replicates (N = 4).

[0249] It can be seen that compared with the model control group (MC), the composition (SP + PI) can significantly reduce the relative expression level of CIRBP, thereby producing an anti-cold-stimulus effect. In addition, the reduction amplitude is more significant than that of the single components of SP and PI, indicating that the combination of SP + PI produces a synergistic effect that is not possessed by the single components when used alone.

[0250] For the tests of the relevant effects involved in Test Examples 7-12, the inventors further tested with the compositions in Examples 1-2 and 4 and found that they also had the same effects.

[0251] The above embodiments are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications made without departing from the spirit and principle of the present invention shall be equivalent replacement methods and are all included in the protection scope of the present invention.

Claims

1. A composition, characterized in that The composition comprises at least two of the following components: Yeast polypeptides, polyamino acid polysaccharide condensates and galactosyl yeast fermentation product filtrates; Preferably, the composition comprises a combination of yeast polypeptides, polyamino acid polysaccharide condensates and galactosyl yeast fermentation product filtrate; or A combination of yeast polypeptides and galactosylceras fermentation filtrate.

2. The composition according to claim 1, characterized in that In the composition, the mass ratio of yeast polypeptides to filtrate of galactosyl yeast-like fermentation product is 0.00001-0.00010:

1.

3. The composition according to claim 1, characterized in that In the composition, the mass ratio of yeast polypeptides, polyamino acid polysaccharide condensation products and galactose yeast-like bacteria fermentation product filtrate is 0.00001-0.00010:0.0001-0.0002:

1.

4. The composition according to claim 1, characterized in that The composition comprises: By mass, 0.0000005-0.00005 parts of yeast polypeptide, 0.000005-0.00005 parts of polyamino acid polysaccharide condensate and 0.01-0.5 parts of galactosyl yeast fermentation product filtrate; or By mass, 0.0000005-0.00005 parts of yeast polypeptide and 0.01-0.5 parts of galactosyl yeast fermentation product filtrate.

5. A method for preparing the composition according to any one of claims 1 to 4, comprising the steps of: The yeast polypeptides, the filtrate of the fermentation product of galactosyl yeast-like bacteria, and the optional polyamino acid polysaccharide condensate are mixed according to the mass ratio described in claim 2 or 3 to obtain the product.

6. A cosmetic, characterized in that: The cosmetic contains the composition according to any one of claims 1 to 4; Preferably, in the cosmetic, based on the total weight of the cosmetic, the composition accounts for 0.1-5%.

7. Use of the composition according to any one of claims 1 to 4 in the preparation of cosmetics or medicines.

8. The use according to claim 7, characterized in that: The cosmetics or medicines also contain cosmetically acceptable excipients or pharmaceutically acceptable excipients.

9. The use according to claim 7, characterized in that: The cosmetic or medicine has at least one of the following functions (1)-(7): (1) Antioxidant; (2) Protect mitochondria; (3) Anti-stress; (4) Resistance to photodamage or photoaging; (5) Anti-inflammatory or inflammatory damage; (6) Promote skin barrier formation or enhance skin barrier defense; (7) Anti-skin irritation.

10. The use according to claim 9, characterized in that: The anti-skin irritation includes: anti-skin cold irritation, anti-skin heat irritation and anti-skin pain and itching irritation.

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