Skin care product containing human-derived fermented polypeptide as well as preparation method and application of skin care product

Abstract

The invention relates to a skin care product containing human-derived fermented polypeptide as well as a preparation method and application of the skin care product, and belongs to the technical field of cosmetics. The skin care product is prepared from the following components in parts by weight: 5 to 8 parts of humanized fermented polypeptide, 10 to 16 parts of purple coneflower extract, 13 to 18 parts of herba portulacae extract and 6 to 10 parts of chamomilla recutita flower extract. Wherein the humanized fermentation polypeptide is prepared from recombinant humanized III-type collagen through composite strain fermentation. All the components in the skin care product have a synergistic effect to form a closed-loop efficacy system of barrier repairing, immunoregulation, relieving and anti-inflammation and moisturizing and water locking, and the skin care product has excellent multiple effects of resisting aging, relieving and repairing, moisturizing and water locking, resisting oxidation, whitening and brightening and the like. Experiments prove that the skin care product has remarkable anti-oxidation, anti-inflammatory and anti-wrinkle properties, is free of irritation and high in biological safety in a human body patch test, and conforms to the modern natural and efficient skin care trend.

Description

Technical Field

[0001] This invention relates to the field of cosmetic technology, specifically to a skin care product containing human-derived fermented polypeptides, its preparation method, and its application. Background Technology

[0002] As consumers' skincare needs become more refined, skincare products with multiple functions such as anti-aging, soothing and repairing, moisturizing and whitening have become the mainstream development direction in the market. Peptide ingredients, due to their high compatibility with human skin and their ability to effectively promote collagen synthesis and improve skin laxity and wrinkles, are widely used in anti-aging skincare products. However, current peptide skincare products on the market generally suffer from two major problems: First, the peptides used are mostly chemically synthesized or directly extracted collagen fragments, with large molecular weights, low skin penetration, and difficulty in fully realizing their biological activity; second, the efficacy of single peptide ingredients is limited, failing to simultaneously meet the comprehensive needs of sensitive skin groups for anti-inflammation, soothing, and barrier repair. Some products also contain irritating additives, easily causing adverse reactions such as skin redness and stinging.

[0003] While recent studies have attempted to enhance the transdermal absorption and bioactivity of active ingredients using microbial fermentation technology, these efforts have generally encountered problems such as arbitrary selection of microbial strains, unclear fermentation substrates, and a wide molecular weight distribution of products, leading to unstable efficacy and significant batch-to-batch variations. Particularly in the fermentation and transformation of human collagen, a key challenge remains: how to precisely control the degree of hydrolysis through specific microbial communities to obtain small-molecule peptides with both high permeability and high bioactivity, and then scientifically combine them with plant ingredients possessing barrier repair, immune regulation, and anti-inflammatory soothing functions to construct a logically closed-loop, synergistic anti-wrinkle system. Therefore, there is an urgent need to develop a novel anti-wrinkle skincare product based on precise fermentation technology, with clearly defined components, a clear mechanism, and high safety and efficacy. Summary of the Invention

[0004] In order to overcome the shortcomings of the existing technology, the present invention aims to provide a skin care product containing human-derived fermented polypeptides, its preparation method and application, wherein the skin care product has excellent anti-wrinkle, soothing and repairing, moisturizing and water-locking, antioxidant and whitening and brightening effects.

[0005] In a first aspect, the present invention provides a skin care product containing human-derived fermented polypeptides, which, by weight, comprises 5-8 parts of human-derived fermented polypeptides, 10-16 parts of purple coneflower extract, 13-18 parts of purslane extract, and 6-10 parts of chamomile extract.

[0006] The human-derived fermented polypeptide is prepared by fermentation of recombinant humanized type III collagen.

[0007] The preparation method of the human fermented polypeptide is as follows: S1: Weigh recombinant humanized type III collagen, add sucrose solution to prepare a liquid fermentation substrate with a collagen concentration of 15-25 g / L; sterilize the fermentation substrate by high pressure steam at 115℃ for 20 min, cool to 37℃±1℃, and set aside.

[0008] S2: Inoculate the sterilized fermentation substrate with a compound microbial strain, wherein the compound microbial strain is prepared by mixing Bifidobacterium lactis BL-99, Lactobacillus rhamnosus GG, and Lactobacillus plantarum LP45 in a live count ratio of 1:2:1.5; the inoculation amount of the compound microbial strain is 7×10⁻⁶. 9 CFU / L-9×10 9 CFU / L; fermentation parameters were: temperature 37℃, initial pH 6.5, dissolved oxygen controlled at 5%-8%, stirring speed 80r / min, fermentation time 48-56h; fermentation broth was obtained after fermentation.

[0009] S3: Centrifuge the fermentation broth, collect the supernatant, ultrafilter the supernatant through a 2000 Da ultrafiltration membrane, collect the filtrate, filter the filtrate through a 0.22 μm filter membrane to obtain sterilized filtrate, freeze-dry the sterilized filtrate to obtain human fermented peptides.

[0010] Secondly, the present invention provides a method for preparing a skin care product containing human-derived fermented polypeptides as described in the first aspect, comprising the following steps: mixing human-derived fermented polypeptides, purple coneflower extract, purslane extract, and chamomile extract according to the formula amount, and sieving to obtain the product.

[0011] Preferably, the sieve is a 100-160 mesh sieve.

[0012] Thirdly, the present invention provides the application of the skin care product containing human-derived fermented polypeptides described in the first aspect in the preparation of cosmetics.

[0013] Preferably, the cosmetic includes any one of serum, gel, hand cream, lotion, ointment or face cream.

[0014] Preferably, the content of human-derived fermented polypeptides in the cosmetics is 0.5wt%-15wt%; more preferably, the content is 1wt%-10wt%.

[0015] Purple coneflower extract: Contains echinacoside, polyphenols, polysaccharides, and other active ingredients, possessing strong anti-inflammatory, antioxidant, antibacterial, and immunomodulatory functions. It can inhibit hyaluronidase activity, reduce inflammatory cell infiltration, and relieve skin redness and allergies; scavenge free radicals and prevent photoaging; inhibit melanin production, and also has a whitening effect.

[0016] Purslane extract: Rich in flavonoids, polyphenols, vitamin C, and alkaloids, it possesses broad-spectrum antibacterial, anti-inflammatory, antioxidant, moisturizing, and wound-healing properties. It can inhibit acne-causing bacteria such as Staphylococcus aureus, reducing acne occurrence; regulate the skin's oil-water balance, enhancing barrier function; inhibit tyrosinase activity, reducing melanin deposition and improving pigmentation.

[0017] Chamomile extract: Contains flavonoids (such as apigenin), amino acids, vitamin A, etc., and has soothing, calming, anti-inflammatory, antioxidant, moisturizing, and epidermal metabolism regulating functions. It can relieve skin stinging and redness, promote cell regeneration, regulate stratum corneum hydration, maintain water-oil balance, and has antioxidant and anti-wrinkle effects, protecting the skin from environmental damage.

[0018] Beneficial effects This invention provides a skin care product containing human-derived fermented polypeptides, its preparation method, and its application. The skin care product has excellent anti-wrinkle, soothing and repairing, moisturizing and water-locking, antioxidant, and whitening and brightening effects.

[0019] This invention utilizes four components—human-derived fermented peptides, purple coneflower extract, purslane extract, and matricaria extract—to form a closed-loop synergistic efficacy system encompassing "barrier repair, immune regulation, soothing and anti-inflammatory, and moisturizing." The human-derived fermented peptides repair the skin barrier and enhance the skin's absorption efficiency of plant extracts, laying the foundation for the active ingredients to exert their effects. Purple coneflower extract exerts immune regulation and anti-inflammatory effects, while matricaria extract targets and soothes sensitivity and promotes cell regeneration. Together, they regulate the skin's microecology, creating a non-inflammatory environment for the cell repair of the human-derived fermented peptides and strengthening anti-aging effects. Purslane extract forms a moisturizing film, combining with the stratum corneum densification effect of the peptides to achieve double moisturization, solving the problem of dry skin after peptide repair. Simultaneously, the immune-enhancing effect of purple coneflower, the targeted soothing effect of matricaria, and the broad-spectrum anti-allergic effect of purslane form a triple anti-inflammatory mechanism, covering various sensitivity triggers. The four components work synergistically to achieve comprehensive skincare effects including anti-aging, soothing and repair, moisturizing and hydrating, and brightening. Attached Figure Description

[0020] Figure 1 Images of human fermented peptides from Examples 1-5 (average molecular weight determined by HPLC-MS: 925.7 Da). Detailed Implementation

[0021] The technical solutions of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. The described embodiments are only some preferred embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0022] Unless otherwise specified, the experimental methods used in the embodiments are conventional or common methods in the art, and the materials and reagents used are commercially available unless otherwise specified.

[0023] The raw materials used in the examples and comparative examples are described below: Purple coneflower extract: purchased from Baoji Liupanyun Biotechnology Co., Ltd.; Purslane extract: purchased from Shaanxi Junhe Biotechnology Co., Ltd.; Matricaria extract: purchased from Lanzhou Waterles Biotechnology Co., Ltd.; Recombinant humanized type III collagen: Guangdong Saiyuan Biomedical Technology Co., Ltd.; Bifidobacterium lactis BL-99: purchased from China General Microbiological Culture Collection Center, accession number: CGMCC NO.15650; Lactobacillus rhamnosus GG: purchased from China General Microbiological Culture Collection Center, accession number: CGMCC NO.1.3724; Lactobacillus plantarum LP45: purchased from China General Microbiological Culture Collection Center, accession number: CGMCC NO.8072; Lactobacillus rhamnosus: purchased from China General Microbiological Culture Collection Center, accession number: CGMCCNO.19510.

[0024] Unless otherwise specified, all components and raw materials used in the embodiments and comparative examples of this invention are commercially available, and the same type of components and raw materials are used in each parallel experiment.

[0025] The preparation method of human-derived fermented polypeptides (self-made) is as follows: S1: Weigh recombinant humanized type III collagen, add it to a sucrose solution with a concentration of 10 g / L and mix well to obtain a liquid fermentation substrate with a collagen concentration of 20 g / L. The liquid fermentation substrate was transferred to a fermentation vessel, sterilized by high-pressure steam at 115℃ for 20 minutes, and cooled to 37℃±1℃ to obtain sterilized fermentation substrate for later use.

[0026] S2: Inoculate the sterilized fermentation substrate with a compound microbial strain, wherein the compound microbial strain is prepared by mixing Bifidobacterium lactis BL-99 (CGMCC NO.15650), Lactobacillus rhamnosus GG (CGMCC NO.1.3724), and Lactobacillus plantarum LP45 (CGMCC NO.8072) in a live count ratio of 1:2:1.5; the inoculation amount of the compound microbial strain is 8 × 10⁻⁶. 9CFU / L; Fermentation parameters: temperature 37℃, initial pH 6.5, dissolved oxygen controlled at 8% (micro-aerobic environment, suitable for the growth of Bifidobacteria), stirring speed 80r / min, fermentation time 48-56h; fermentation broth obtained after fermentation.

[0027] S3: Centrifuge the fermentation broth at 8000g for 20min, collect the supernatant, ultrafilter the supernatant through a 2000 Da ultrafiltration membrane, collect the filtrate, filter the filtrate through a 0.22μm filter membrane to obtain sterilized filtrate, freeze-dry the sterilized filtrate to obtain human fermented peptide (average molecular weight determined by HPLC-MS: 925.7Da), used in each example and comparative examples 1-4.

[0028] Example 1 A skincare product containing human-derived fermented peptides, by weight, comprises 7 parts human-derived fermented peptides, 13 parts purple coneflower extract, 15 parts purslane extract, and 8 parts matricaria extract, with a total mass of 60g.

[0029] The preparation method of the skin care product containing human-derived fermented polypeptides includes the following steps: mixing human-derived fermented polypeptides, purple coneflower extract, purslane extract, and chamomile extract according to the formula amount, and passing them through a 120-mesh sieve to obtain the product.

[0030] Example 2 A skincare product containing human-derived fermented peptides, by weight, comprises 5 parts human-derived fermented peptides, 13 parts purple coneflower extract, 14 parts purslane extract, and 6 parts matricaria extract, with a total mass of 60g.

[0031] The preparation method of the skin care product containing human-derived fermented polypeptides is the same as that in Example 1.

[0032] Example 3 A skincare product containing human-derived fermented peptides, by weight, comprises 7 parts human-derived fermented peptides, 10 parts purple coneflower extract, 16 parts purslane extract, and 10 parts matricaria extract, with a total mass of 60g.

[0033] The preparation method of the skin care product containing human-derived fermented polypeptides is the same as that in Example 1.

[0034] Example 4 A skincare product containing human-derived fermented peptides, by weight, comprises 6 parts human-derived fermented peptides, 15 parts purple coneflower extract, 18 parts purslane extract, and 9 parts matricaria extract, with a total mass of 60g.

[0035] The preparation method of the skin care product containing human-derived fermented polypeptides is the same as that in Example 1.

[0036] Example 5 A skincare product containing human-derived fermented peptides, by weight, comprises 8 parts human-derived fermented peptides, 15 parts purple coneflower extract, 15 parts purslane extract, and 8 parts matricaria extract, with a total mass of 60g.

[0037] The preparation method of the skin care product containing human-derived fermented polypeptides is the same as that in Example 1.

[0038] Comparative Example 1 A skincare product, by weight, contains 13 parts of purple coneflower extract, 15 parts of purslane extract, and 8 parts of matricaria extract, for a total weight of 60g.

[0039] The preparation method of the skin care product is the same as that in Example 1, except that the human-derived fermented polypeptide component is missing.

[0040] Comparative Example 2 A skincare product, by weight, contains 7 parts human-derived fermented polypeptides, 15 parts purslane extract, and 8 parts chamomile extract, with a total mass of 60g.

[0041] The preparation method of the skin care product is the same as that in Example 1, except that the component purple coneflower extract is missing.

[0042] Comparative Example 3 A skincare product, by weight, contains 7 parts of human-derived fermented polypeptide, 13 parts of purple coneflower extract, and 8 parts of chamomile extract, with a total mass of 60g.

[0043] The preparation method of the skin care product is the same as that in Example 1, except that the purslane extract component is missing.

[0044] Comparative Example 4 A skincare product, by weight, contains 7 parts of human-derived fermented polypeptide, 13 parts of purple coneflower extract, and 15 parts of purslane extract, with a total weight of 60g.

[0045] The preparation method of the skin care product is the same as that in Example 1, except that the chamomile extract component is missing.

[0046] Comparative Example 5 A skincare product, by weight, contains 7 parts of recombinant humanized type III collagen, 13 parts of purple coneflower extract, 15 parts of purslane extract, and 8 parts of matricaria extract, for a total weight of 60g.

[0047] The preparation method of the skin care product is the same as that in Example 1, except that the human fermented peptides are replaced with an equal amount of recombinant humanized type III collagen.

[0048] Comparative Example 6 A skincare product, by weight, comprises 7 parts of human-derived fermented polypeptide, 13 parts of purple coneflower extract, 15 parts of purslane extract, and 8 parts of chamomile extract, with a total mass of 60g. The preparation method of the skincare product is the same as that in Example 1.

[0049] The preparation method of the human fermented polypeptide in Comparative Example 6 differs from that in Example 1, except that the "compound strain is obtained by combining *Lactobacillus rhamnosus* GG and *Lactobacillus plantarum* LP45 in a viable ratio of 2:1.5". The preservation numbers for *Lactobacillus rhamnosus* GG and *Lactobacillus plantarum* LP45 are CGMCC NO. 1.3724 and CGMCC NO. 8072, respectively. The average molecular weight of the human fermented polypeptide prepared in Comparative Example 6 was determined to be 1428.3 Da by HPLC-MS.

[0050] Comparative Example 7 A skincare product, by weight, comprises 7 parts of human-derived fermented polypeptide, 13 parts of purple coneflower extract, 15 parts of purslane extract, and 8 parts of chamomile extract, with a total mass of 60g. The preparation method of the skincare product is the same as that in Example 1.

[0051] The preparation method of the human fermented polypeptide in Comparative Example 7 differs from that in Example 1, except that the "compound strain was obtained by combining Bifidobacterium lactis BL-99 and Lactobacillus rhamnosus GG in a live count ratio of 1:2". The preservation numbers for Bifidobacterium lactis BL-99 and Lactobacillus rhamnosus GG are CGMCC NO.15650 and CGMCC NO.1.3724, respectively. The average molecular weight of the human fermented polypeptide prepared in Comparative Example 7 was determined to be 1379.5 Da by HPLC-MS.

[0052] Comparative Example 8 A skincare product, by weight, comprises 7 parts of human-derived fermented polypeptide, 13 parts of purple coneflower extract, 15 parts of purslane extract, and 8 parts of chamomile extract, with a total mass of 60g. The preparation method of the skincare product is the same as that in Example 1.

[0053] The preparation method of the human fermented polypeptide in Comparative Example 8 differs from that in Example 1, except that the complex bacterial strain is prepared by combining Bifidobacterium lactis BL-99 (accession number: CGMCC NO.15650), Lactobacillus rhamnosus (accession number: CGMCC NO.19510), and Lactobacillus plantarum LP45 (accession number: CGMCC NO.8072) in a ratio of 1:2:1.5, that is, Lactobacillus rhamnosus GG (accession number: CGMCC NO.1.3724) is replaced with Lactobacillus rhamnosus (accession number: CGMCC NO.19510); the average molecular weight of the human fermented polypeptide prepared in Comparative Example 8 was determined by HPLC-MS to be 1153.8 Da.

[0054] Table 1. Raw material formulations (parts by weight) for Examples 1-5 and Comparative Examples 1-8 Human-derived fermented peptides Purple coneflower extract Purslane extract Matricaria extract Recombinant humanized type III collagen Example 1 7 13 15 8 / Example 2 5 13 14 6 / Example 3 7 10 16 10 / Example 4 6 15 18 9 / Example 5 8 15 15 8 / Comparative Example 1 / 13 15 8 / Comparative Example 2 7 / 15 8 / Comparative Example 3 7 13 / 8 / Comparative Example 4 7 13 15 / / Comparative Example 5 / 13 15 8 7 Comparative Example 6 7 13 15 8 / Comparative Example 7 7 13 15 8 / Comparative Example 8 7 13 15 8 / Note: " / " indicates that the component was not added.

[0055] Performance verification tests of the examples and comparative examples.

[0056] 1. DPPH scavenging rate test: DPPH scavenging rate test was conducted on Examples 1-5 and Comparative Examples 1-8 to determine the scavenging ability of the test samples on 1,1-diphenyl-2-trinitrophenylhydrazine (DPPH) free radicals, and to evaluate the antioxidant activity of the skin care products prepared in Examples 1-5 and Comparative Examples 1-8.

[0057] Using deionized water as a solvent, the anti-aging compositions prepared in Examples 1-5 and Comparative Examples 1-8 were formulated into homogeneous sample solutions with a concentration of 0.1 mg / mL, which were then used as test sample solutions for later use.

[0058] Sample group (sample 1): Take 3 mL of sample with a concentration of 2×10 4 Mix 3 mL of the above-mentioned sample solution with anhydrous DPPH ethanol solution at a concentration of mol / L and shake well.

[0059] Blank control group (sample 2): Take 3 mL of sample with a concentration of 2×10 4 Mix 3 mL of deionized water with anhydrous DPPH ethanol solution at mol / L and shake well (to determine the initial absorbance of undeletable DPPH free radicals).

[0060] Sample blank group (sample 3): Take 3 mL of anhydrous ethanol and mix it thoroughly with 3 mL of the above sample solution. Shake well (to eliminate the interference of absorbance of the sample itself and the solvent at the measurement wavelength).

[0061] Samples 1, 2, and 3 were placed in the dark for 30 minutes, and the absorbance of the mixed solution at 517 nm was measured.

[0062] Calculation: DPPH clearance rate = [1 - (A1 - A3) / A2] × 100%.

[0063] 2. Anti-inflammatory performance test: The TNF-α inhibition rate was measured. The test method is as follows: (1) Seeding: Logarithmic growth phase RAW264.7 cells were seeded into culture flasks and 24-well culture plates at a density of 1×10⁻⁶ cells / well. 6 The cells / mL were incubated in DMEM medium at 37°C for 24 hours in a 5% CO2 incubator. (2) Model making: Control group: Remove the supernatant from the 24-well plate, repeat in parallel with 5 wells, and add 0.5 μL of DMEM culture medium and 0.1 μL of PBS solution to each well; LPS model group: Remove the supernatant from the 24-well plate, add 500 μL of DMEM culture medium and 100 μL of LPS solution to each of the 5 parallel wells.

[0064] Experimental group: Remove the supernatant from the 24-well plate, add 500 μL of DMEM culture medium, 100 μL of LPS and a mixed solution of sample (example and comparative sample) to each of the 5 parallel wells.

[0065] (3) Detection: After incubation at 37℃ in a 5% CO2 incubator for 24 hours, the supernatant was collected. The sample was centrifuged in a centrifuge tube, and the supernatant was used for the detection of inflammatory factor concentration. Calculation: Inflammatory factor inhibition rate (%) = [(average content of inflammatory factors in the model group - average content of inflammatory factors in the treatment group) / (average content of inflammatory factors in the model group - average content of inflammatory factors in the blank control group)] × 100%.

[0066] Table 2. Record of DPPH clearance rate (%) and TNF-α inhibition rate (%) Group DPPH free radical scavenging rate (%) TNF-α inhibition rate (%) Example 1 95.3% 91.1% Example 2 92.6% 89.2% Example 3 94.1% 87.3% Example 4 89.2% 89.7% Example 5 91.7% 88.5% Comparative Example 1 76.3% 81.3% Comparative Example 2 65.7% 61.1% Comparative Example 3 74.6% 67.9% Comparative Example 4 68.0% 71.2% Comparative Example 5 65.7% 69.3% Comparative Example 6 77.2% 76.4% Comparative Example 7 76.6% 78.5% Comparative Example 8 81.5% 79.4% According to the data in Table 2: 1) The DPPH free radical scavenging rate of Examples 1-5 ranged from 89.2% to 95.3%, and the TNF-α inhibition rate ranged from 87.3% to 91.1%. Both indicators were significantly better than all comparative examples, indicating that Examples 1-5 have good antioxidant and anti-inflammatory effects. 2) Compared with Example 1, Comparative Examples 1-4 lacked one component from human fermented peptides, purple coneflower extract, purslane extract, and magnolia extract, respectively, resulting in a significant decrease in DPPH free radical scavenging rate (65.7%-76.3%) and TNF-α inhibition rate (61.1%-81.3%). This result proves that the combined formulation of human fermented peptides, purple coneflower extract, purslane extract, and magnolia extract can produce a synergistic effect, achieving highly efficient antioxidant and anti-inflammatory effects. 3) Compared with Example 1, Comparative Example 5 replaced the human fermented peptides with an equal amount of recombinant humanized type III collagen. The DPPH free radical scavenging rate and TNF-α inhibition rate decreased to 65.7% and 69.3%, respectively. This indicates that the fermentation process can significantly enhance the activity of collagen, and the fermented peptides are more easily utilized by cells. 4) Compared with Example 1, the composition of the compound strains in the preparation of human fermented peptides in Comparative Examples 6-8 was changed. Comparative Example 6 lacked Bifidobacterium lactis BL-99 (accession number: CGMCC NO.15650), Comparative Example 7 lacked Lactobacillus plantarum LP45 (accession number: CGMCC NO.8072), and Comparative Example 8 replaced Lactobacillus rhamnosus GG (accession number: CGMCC NO.1.3724) with Lactobacillus rhamnosus (accession number: CGMCC NO.19510). This resulted in a significant decrease in the DPPH free radical scavenging rate and TNF-α inhibition rate of Comparative Examples 6-8. This indicates that Bifidobacterium lactis BL-99, Lactobacillus rhamnosus GG, and Lactobacillus plantarum LP45 can synergistically ferment recombinant humanized type III collagen and prepare human fermented peptides with higher activity when combined in a specific ratio.

[0067] Application examples Application Examples 1-5 and Comparative Application Examples 1-8 of this invention provide a whitening and anti-wrinkle skin care lotion. The ingredients (wt%) of the whitening and anti-wrinkle skin care lotion are shown in Table 3. In application examples 1-5 and comparative application examples 1-8, the skin care products prepared in examples 1-5 and comparative examples 1-8 are used respectively.

[0068] Table 3. Ingredients of Whitening and Anti-Wrinkle Skin Lotion Raw material name Percentage by weight (wt%) The skincare products described in Examples 1-5 or Comparative Examples 1-8 3.50 Carbomer 1.00 glycerin 1.60 Cetearyl alcohol polyether-20 1.50 Polyglycerol-3-diisostearate 2.00 PEG800 0.20 Panthenol 0.80 Trehalose 1.50 Shea butter 0.50 Arginine 0.03 Phenoxyethanol 0.25 Ethylhexylglycerin 0.20 sterile deionized water margin The preparation method of the whitening and anti-wrinkle skin care lotion includes the following steps: Step 1: Mix glycerin, cetearyl alcohol polyether-20, polyglycerol-3-diisostearate, and shea butter, and heat at 55°C until completely dissolved to obtain the oil phase.

[0069] Step 2: Heat sterile deionized water to 55°C, add carbomer and trehalose, and stir until completely dissolved (the amount of sterile deionized water is 10 times the total mass of carbomer and trehalose); then add PEG800, panthenol, arginine and the skin care product provided in the examples or comparative examples, and stir thoroughly until completely dissolved to obtain the aqueous phase.

[0070] Step 3: Under 55℃ water bath conditions, add the oil phase to the aqueous phase and shear and stir at 3000 rpm for 5 minutes using a high-speed homogenizer to obtain the primary emulsion. Add phenoxyethanol and ethylhexylglycerin to the remaining water and stir evenly. Then add the primary emulsion to the mixture to obtain a mixture. Homogenize the mixture under high pressure at 70 MPa for 3 cycles to obtain the final product.

[0071] The whitening and anti-wrinkle skin lotions prepared in Application Examples 1-5 and Comparative Application Examples 1-8 were subjected to the following performance tests.

[0072] 1. Human closed patch test: The test is conducted using human skin patch test. The specific test method is in accordance with the "Cosmetic Safety Technical Specifications" (2015 edition), see Table 4 for details.

[0073] Table 4 Evaluation Table of Human Skin Stimulation degree of reaction Rating levels Skin reaction － 0 negative reaction ± 1 Suspicious reaction, only slight erythema ＋ 2 Weak positive reaction (erythema): erythema, infiltration, edema, and papules may be present. ＋＋ 3 Strong positive reaction (herpes): erythema, infiltration, edema, papules, vesicles; the reaction may extend beyond the test area. ＋＋＋ 4 Extremely strong positive reaction: obvious erythema, severe infiltration, edema, confluent herpes, reaction extending beyond the test area. 130 participants were recruited, underwent ethical review, and signed informed consent forms. The average age of the participants was 24.3 years. The participants were randomly divided into 13 groups, and 25 μL of samples from Examples 1-5 and Comparative Examples 1-8 were used in the experiment. The experimental results are recorded in Table 5.

[0074] Table 5. Results of patch test Number of people in the project negative reaction Suspicious reaction Weak positive reaction Strong positive reaction Extremely strong positive reaction Application Example 1 10 0 0 0 0 Application Example 2 10 0 0 0 0 Application Example 3 10 0 0 0 0 Application Example 4 10 0 0 0 0 Application Example 5 10 0 0 0 0 Comparative Application Example 1 10 0 0 0 0 Comparative Application Example 2 10 0 0 0 0 Comparative Application Example 3 10 0 0 0 0 Comparative Application Example 4 10 0 0 0 0 Comparative Application Example 5 10 0 0 0 0 Comparative Application Example 6 10 0 0 0 0 Comparative Application Example 7 10 0 0 0 0 Comparative Application Example 8 10 0 0 0 0 According to the data in Table 5, the subjects in Application Examples 1-5 and Comparative Application Examples 1-8 all had negative skin reactions, and no adverse skin irritation reactions such as erythema or herpes occurred. This indicates that the whitening and anti-wrinkle skin lotion prepared using Application Examples 1-5 (using the skin care products of Examples 1-5) has a soothing and gentle effect and high biosafety, providing effective experimental data support for the application of the skin care products described in Examples 1-5 of this invention in the preparation of cosmetics.

[0075] 11. Anti-wrinkle test This study recruits 130 female participants aged 28-53. Inclusion criteria include: visible wrinkles around the eyes and forehead (wrinkle depth not less than 0.1 mm); no active skin lesions (including but not limited to eczema, contact dermatitis, acne, etc.); no known allergies or history of drug / cosmetic allergies; no topical or systemic use of skincare products containing anti-wrinkle active ingredients such as retinol and its derivatives, bioactive peptides, or high concentrations of ascorbic acid within the past three months; and no prior medical aesthetic wrinkle removal interventions (including but not limited to botulinum toxin type A injections, intense pulsed light therapy, radiofrequency skin tightening, etc.). All participants must sign a written informed consent form, confirming their voluntary participation and commitment to fully cooperate with the study process.

[0076] Subjects were randomly assigned to 13 experimental units using a random number table, with 10 subjects in each unit, corresponding to the test samples prepared in Application Examples 1-5 and Control Examples 1-8, respectively. Subjects applied the designated sample to the target wrinkle areas (around the eyes and forehead) daily after facial cleansing in the morning and evening, for a total intervention period of 28 days.

[0077] At the baseline period (D0) and the end of the intervention (D... 28 The Antera 3D CS three-dimensional skin imaging analysis system was used to perform non-invasive skin morphology detection on a designated area around the eyes of the subjects (scanning area 8mm × 8mm) to obtain quantitative data on wrinkle depth. The degree of wrinkle improvement was calculated using the following formula: Wrinkle improvement rate (%) = (D0 - D) 28 ) / D0×100%, where D0 represents the baseline wrinkle depth, D 28 This represents the depth of wrinkles after 28 days of intervention. The detection sites for each subject were measured three times, and the arithmetic mean was included in the statistical analysis.

[0078] Table 6. Wrinkle depth reduction rate (%) Group Wrinkle depth reduction rate (%) Application Example 1 33.7% Application Example 2 31.8% Application Example 3 30.6% Application Example 4 29.4% Application Example 5 31.2% Comparative Application Example 1 18.3% Comparative Application Example 2 15.3% Comparative Application Example 3 17.9% Comparative Application Example 4 16.5% Comparative Application Example 5 14.2% Comparative Application Example 6 22.1% Comparative Application Example 7 23.4% Comparative Application Example 8 24.3% According to the data in Table 6, the wrinkle depth reduction rate of Application Examples 1-5 ranged from 29.4% to 33.7%. In contrast, the reduction rates of Application Examples 1-8 were all below 25%, with Application Example 1, lacking the human-derived fermented peptide, showing a reduction rate of only 18.3%, and Application Example 5, which replaced the human-derived fermented peptide, showing the lowest reduction rate at only 14.2%. Furthermore, Application Examples 2-4, lacking Echinacea purpurea, Portulaca oleracea, and Matricaria champaca extracts respectively, showed a wrinkle depth reduction rate of only 15.3%-17.9%, significantly lower than Application Example 1. This indicates that the synergistic effect of the human-derived fermented peptide and the three plant extracts (Echinacea purpurea, Portulaca oleracea, and Matricaria champaca extracts) in this invention can effectively promote collagen synthesis in the skin and improve wrinkles and sagging.

[0079] Compared to Application Examples 6-8, which had altered fermentation strain formulations, the prepared peptides had larger molecular weights and lower activity, resulting in wrinkle depth reduction rates of 22.1%-24.3%, lower than Application Example 1. This demonstrates that the combined fermentation of Bifidobacterium lactis BL-99, Lactobacillus rhamnosus GG, and Lactobacillus plantarum LP45 in a 1:2:1.5 ratio (live count ratio) in this invention is necessary to prepare peptides with smaller molecular weights and higher activity, ensuring the anti-wrinkle effect.

[0080] The preferred embodiments of the present invention disclosed above are merely illustrative of the invention. These preferred embodiments do not exhaustively describe all details, nor do they limit the invention to the specific implementations described. Clearly, many modifications and variations can be made based on the content of this specification. This specification selects and specifically describes these embodiments to better explain the principles and practical applications of the invention, thereby enabling those skilled in the art to better understand and utilize the invention. The invention is limited only by the claims and their full scope and equivalents.

Claims

1. A skincare product containing human-derived fermented polypeptides, characterized in that, By weight, it contains 5-8 parts of human-derived fermented polypeptide, 10-16 parts of purple coneflower extract, 13-18 parts of purslane extract, and 6-10 parts of matricaria extract.

2. The skincare product containing human-derived fermented polypeptides according to claim 1, characterized in that, The human fermented polypeptide is prepared by fermentation of recombinant humanized type III collagen.

3. The skincare product containing human-derived fermented polypeptides according to claim 2, characterized in that, The preparation method of the human fermented polypeptide is as follows: S1: Weigh recombinant humanized type III collagen, add sucrose solution to prepare a liquid fermentation substrate with a collagen concentration of 15-25 g / L, and sterilize it for later use; S2: Inoculate the sterilized fermentation substrate with a compound microbial strain, wherein the inoculation amount of the compound microbial strain is 7 × 10⁻⁶. 9 CFU / L-9×10 9 CFU / L; fermentation conditions were: temperature 37℃, initial pH 6.5, dissolved oxygen controlled at 5%-8%, stirring speed 80r / min, fermentation time 48-56h; fermentation broth was obtained after fermentation. S3: Centrifuge the fermentation broth, collect the supernatant, ultrafilter the supernatant through a 2000 Da ultrafiltration membrane, collect the filtrate, filter the filtrate through a 0.22 μm filter membrane to obtain sterilized filtrate, freeze-dry the sterilized filtrate to obtain human fermented peptides.

4. The skincare product containing human-derived fermented polypeptides according to claim 3, characterized in that, The compound bacterial strain was prepared by mixing Bifidobacterium lactis BL-99, Lactobacillus rhamnosus GG, and Lactobacillus plantarum LP45 in a live count ratio of 1:2:1.5; the preservation number of Bifidobacterium lactis BL-99 is CGMCC NO.15650, the preservation number of Lactobacillus rhamnosus GG is CGMCC NO.1.3724, and the preservation number of Lactobacillus plantarum LP45 is CGMCC NO.8072.

5. The method for preparing a skincare product containing human-derived fermented polypeptides according to any one of claims 1-4, characterized in that, The process includes the following steps: mixing human-derived fermented peptides, purple coneflower extract, purslane extract, and magnolia extract according to the formula, and then sieving to obtain the final product.

6. The preparation method according to claim 5, characterized in that, The sieve is a 100-160 mesh sieve.

7. The use of the skin care product containing human-derived fermented polypeptides according to any one of claims 1-4 in the preparation of cosmetics.

8. The application according to claim 7, characterized in that, The cosmetic product is selected from any one of the following: serum, gel, hand cream, lotion, ointment, and face cream.

9. The application according to claim 7, characterized in that, The cosmetic product containing human-derived fermented polypeptides has a content of 0.5wt%-15wt%.

10. The application according to claim 9, characterized in that, The content is 1wt%-10wt%.

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