A multiple protein synergistic firming and anti-wrinkle soothing and repairing cosmetic composition and a preparation method and application thereof

Abstract

This invention belongs to the field of cosmetic technology, specifically disclosing a multi-protein synergistic firming, anti-wrinkle, soothing, and repairing cosmetic composition, its preparation method, and its application. The cosmetic composition, by weight, is prepared from the following raw materials: 0.001-0.1 parts of Ma Yinglong Babao raw powder, 0.01-5 parts of oat firming protein, 0.01-1 part of silk protein, 0.001-0.1 parts of recombinant collagen, 0.001-0.1 parts of recombinant fibronectin, and 0.0001-0.1 parts of recombinant mussel protein. This invention utilizes the precise combination of oat firming protein, silk protein, recombinant collagen, recombinant fibronectin, recombinant mussel protein, and Ma Yinglong Babao raw powder to construct a four-in-one efficacy system of "firming-anti-wrinkle-soothing-repairing," achieving synergistic effects among the proteins. This solves the problems of existing products having single efficacy, poor synergy, and easy skin irritation, providing a highly effective and gentle comprehensive care solution for aging skin, sensitive skin, and skin with damaged skin barriers.

Description

Technical Field

[0001] This invention belongs to the field of cosmetic technology, specifically relating to a multi-protein synergistic firming, anti-wrinkle, soothing and repairing cosmetic composition, its preparation method and application. Background Technology

[0002] As skincare needs become more refined, consumers' demands for cosmetic efficacy have shifted from a single anti-aging or single repair function to a dual or even multiple demand for "firming and anti-wrinkle + soothing and repairing". This is especially true for people with aging and sensitive skin, whose skin suffers from aging problems such as collagen loss, decreased elasticity, and wrinkle formation, as well as sensitivity issues such as a fragile barrier, redness, and stinging. Conventional cosmetics cannot simultaneously address all four core functions and are prone to the contradiction of "anti-wrinkle ingredients irritating the skin and repairing ingredients weakening anti-aging".

[0003] Currently, most protein-containing cosmetics on the market use a single protein or a simple combination of two proteins, which has obvious technical limitations: First, the effects of a single protein are one-sided. For example, oat firming protein focuses on soothing and moisturizing, silk protein focuses on repairing and nourishing, recombinant collagen focuses on anti-wrinkle and firming, and fibronectin and mussel protein focus on repairing the skin barrier. Using them alone cannot achieve the four effects of firming, anti-wrinkle, soothing, and repairing. Second, existing multi-protein compound products are just simple mixtures of ingredients, without exploring the synergistic effects between the proteins. There are even problems of mutual inhibition of activity, resulting in poor cumulative effects and failing to break through the "1+1<2" dilemma. Third, most products only focus on a single combination of "anti-wrinkle + firming" or "soothing + repairing", failing to achieve the simultaneous achievement of four effects, and ignoring the compatibility of anti-aging ingredients and repairing ingredients. This can easily irritate sensitive skin and cannot meet the needs of multiple skin types.

[0004] In existing technologies, oat firming protein, fibroin, recombinant collagen, fibronectin, and mussel protein are all used individually in the cosmetic field. Oat firming protein can achieve soothing, anti-inflammatory, and barrier repair effects by upregulating the expression of key barrier proteins such as filaggrin and lipofuscin, while also possessing some moisturizing and antioxidant capabilities, but lacking anti-wrinkle and firming effects. Filaggrin can combat fibroblast aging, restore the homeostasis of the skin's extracellular matrix (ECM), promote collagen synthesis, and inhibit collagen degradation, showing anti-wrinkle and repair potential, but its soothing effect is weak. Recombinant collagen, when used alone, can only replenish collagen and improve sagging wrinkles, but cannot solve skin sensitivity issues. Filaggrin can promote cell repair and strengthen the skin barrier, while mussel protein can quickly form a protective film and soothe redness and irritation, but neither of them lacks the core anti-wrinkle and firming effects. Furthermore, existing technologies have not found that the combination of these five proteins can produce a synergistic effect, nor have they achieved a simultaneous enhancement of four effects, failing to meet consumers' demands for highly effective, gentle, and multifunctional cosmetics.

[0005] In view of this, the present invention is proposed. Summary of the Invention

[0006] To address the aforementioned technical problems, the present invention aims to provide a multi-protein synergistic firming, anti-wrinkle, soothing, and repairing cosmetic composition, its preparation method, and its application. The cosmetic composition can achieve multiple effects through synergistic enhancement.

[0007] To achieve the above objectives, the present invention adopts the following technical solution: In a first aspect, the present invention provides a multi-protein synergistic firming, anti-wrinkle, soothing and repairing cosmetic composition, which is prepared by weight from the following raw materials: 0.001~0.1 parts of Mayinglong Babao raw powder, 0.01~5 parts of oat firming protein, 0.01~1 parts of silk protein, 0.001~0.1 parts of recombinant collagen, 0.001~0.1 parts of recombinant fibronectin, and 0.0001~0.1 parts of recombinant mussel protein.

[0008] The above technical solution utilizes a precise blend of oat firming protein, silk protein, recombinant collagen, recombinant fibronectin, recombinant mussel protein, and Ma Yinglong Babao powder to construct a four-in-one efficacy system of "firming, anti-wrinkle, soothing, and repairing." This system achieves synergistic effects among the proteins, addressing the issues of existing products having single efficacy, poor synergy, and easy skin irritation. It provides a highly efficient and gentle comprehensive care solution for aging skin, sensitive skin, and skin with a damaged barrier.

[0009] As a preferred embodiment of the present invention, the cosmetic composition is prepared by weight from the following raw materials: 0.005-0.05 parts of Mayinglong Babao raw powder, 0.1-4 parts of oat firming protein, 0.01-0.5 parts of silk protein, 0.005-0.05 parts of recombinant collagen, 0.005-0.05 parts of recombinant fibronectin, and 0.0005-0.05 parts of recombinant mussel protein.

[0010] As a preferred embodiment of the technical solution of the present invention, the cosmetic composition is prepared by weight from the following raw materials: 0.01 parts of Mayinglong Babao raw powder, 3 parts of oat firming protein, 0.1 parts of silk protein, 0.01 parts of recombinant collagen, 0.01 parts of recombinant fibronectin, and 0.001 parts of recombinant mussel protein.

[0011] As a preferred embodiment of the technical solution of the present invention, the cosmetic composition is prepared by weight of the following raw materials: 2.8 parts oat firming protein, 0.2 parts silk protein, 0.02 parts recombinant collagen, 0.005 parts recombinant fibronectin, and 0.002 parts recombinant mussel protein.

[0012] For example, the cosmetic composition is prepared by weight of the following raw materials: 0.015 parts of Mayinglong Babao raw powder, 3.2 parts of oat firming protein, 0.15 parts of silk protein, 0.03 parts of recombinant collagen, 0.004 parts of recombinant fibronectin, and 0.0015 parts of recombinant mussel protein.

[0013] In the above technical solution, preferably, the recombinant collagen is a three-component recombinant collagen, which is prepared by compounding type I, type III and type XVII recombinant collagen.

[0014] In the above technical solution, preferably, the silk core protein is a silk core protein oil sponge loaded with meadowfoam seed oil, and more preferably, silk core protein oil sponge-meadowfoam seed oil 40.

[0015] In the above technical solution, preferably, the Mayinglong Babao raw powder is obtained by mixing zinc carbonate hydroxide, amber powder and pearl powder in a mass ratio of 80-100:0.1-2:0.1-2; more preferably, the mass ratio of zinc carbonate hydroxide, amber powder and pearl powder is 90-100:0.1-1:0.5-2.

[0016] Secondly, this invention provides the application of the above-mentioned cosmetic composition in the preparation of cosmetics. The cosmetics can be lotions, creams, serums, toners, etc. The aforementioned cosmetic composition can be used to prepare firming, anti-wrinkle, and soothing repair skincare products, especially suitable for daily care of aging sensitive skin, dry and dehydrated skin, and skin with damaged skin barriers. It can effectively improve skin laxity, wrinkles, redness, stinging, and dryness, achieving firm, elastic, and gently repairing effects. It can also be used in post-medical aesthetic repair products to alleviate post-operative skin redness and sensitivity, accelerate skin barrier repair, and broaden the application scenarios of protein-based cosmetics.

[0017] Thirdly, the present invention also provides an essence lotion based on the aforementioned cosmetic composition, comprising the aforementioned cosmetic composition and excipients.

[0018] As a preferred embodiment of the present invention, the excipients are selected from one or more of the following: humectants, soothing agents, antioxidants, oils, water, chelating agents, emulsifiers, preservatives, polyols, and thickeners.

[0019] As a preferred embodiment of the present invention, the moisturizer is sodium hyaluronate; The soothing agent is one or two of dipotassium glycyrrhizate and bisabolol; The polyol is selected from one or more of glycerol, butylene glycol, sorbitol, dipropylene glycol, and 1,3-propanediol; The oil is selected from one or more of the following: polydimethylsiloxane, polydimethylsiloxane alcohol, cyclopentadimethylsiloxane, polydimethylsiloxane cross-linked polymer, hydrogenated polyisobutylene, isopropyl myristate, caprylic / capric triglyceride, isononyl isononanoate, neopentyl glycol dicaprylate / dicaprylate, squalane, and shea butter. The chelating agent is selected from one or two of disodium EDTA and octanoyl hydroxamic acid; The emulsifier is selected from one or more of isocetyl palmitate, cetearyl glucoside, cetearyl alcohol, PEG-100 stearate, cetearyl oleate, and sorbitan oleate; The antioxidant is selected from one or two of tocopherol and tocopherol acetate; The preservative is selected from one or more of phenoxyethanol, ethylhexylglycerin, chlorphenesin, methylparaben, p-hydroxyacetophenone, propylparaben, 1,2-hexanediol, and sorbitan octanoate; The thickener is selected from one or more of the following: polyacrylate crosspolymer-6, hydroxyethyl acrylate / sodium acryloyldimethyl taurate copolymer, sorbitan isostearate, sodium polyacryloyldimethyl taurate, ammonium acryloyldimethyl taurate / VP copolymer, sodium polyacrylate, polyacrylamide, acrylate / C10-30 alkanol acrylate crosspolymer, and carbomer.

[0020] Fourthly, the present invention also provides a method for preparing the above-mentioned essence lotion, comprising the following steps: (1) Add water, chelating agent, moisturizer, first part soothing agent and first part polyol to emulsification tank, stir and heat to 80-85℃ to obtain material A; (2) Add the oil, emulsifier, antioxidant, Mayinglong Babao powder, second part of the soothing agent, and first part of the preservative into the pretreatment tank, stir and heat to 80-85℃ to obtain material B; (3) Add material B to material A, and stir homogenously to obtain material C; (4) Add the thickener and the second part of the polyol to material C, stir homogenize and cool to 40~45℃ to obtain material D; (5) Mix oat firming protein, silk core protein, recombinant collagen, recombinant fibronectin and recombinant mussel protein and add to material D. Homogenize and stir evenly to obtain material E. (6) Add the second part of the preservative to material E, stir homogenously until uniform, and cool to room temperature.

[0021] The above technical solution effectively preserves the activity of the core protein through the process design of "gradient cooling + stirring" while ensuring that the five proteins are evenly dispersed and give full play to their synergistic effect. This is different from the problem of protein inactivation and poor synergy caused by "high temperature and protein addition" in the existing preparation process, and further ensures the efficacy and stability of the product.

[0022] Compared with the prior art, the present invention has the following beneficial effects: (1) This invention uses a precise combination of three types of recombinant collagen (type I, type III, and type XVII) to break through the limitations of single collagen in anti-wrinkle treatment and lay the core foundation for firming and anti-wrinkle treatment: Type I collagen can replenish collagen loss in the dermis, enhance skin support, improve skin laxity and sagging, reduce static wrinkle formation, and provide structural support for the skin; Type III collagen can promote fibroblast proliferation, increase skin elasticity, lighten dynamic wrinkles, make the skin softer, and at the same time assist in repairing the skin barrier, making up for the shortcomings of single type I collagen which only has anti-wrinkle effect but does not repair; Type XVII collagen can anchor the connection between the epidermis and dermis, strengthen the stability of skin structure, reduce collagen loss, and at the same time promote skin barrier repair, achieving a double foundation of "anti-wrinkle + repair". After the three are combined, the anti-wrinkle effect is significantly improved, and the skin barrier function is significantly enhanced, solving the pain point of traditional collagen which only has anti-wrinkle effect but does not repair, and laying the core foundation for the realization of four effects.

[0023] (2) This invention reduces the potential irritation of ingredients such as recombinant collagen and recombinant fibronectin through the synergistic nourishment of oat firming protein and silk core protein. The product's pH value is close to the skin's physiological pH value, making it suitable for various skin types such as sensitive skin, aging skin, and dry skin, and it can be used for a long time. At the same time, it can be applied to post-medical aesthetic repair, broadening the application scenarios and solving the problems of strong irritation and poor compatibility of existing protein cosmetics. Through the synergistic effect of multiple proteins and the optimization of the preparation process, the activity of the core protein can be retained for a long time, and a long-lasting sustained release can be achieved.

[0024] (3) This invention achieves a dual breakthrough of "rapid soothing + long-lasting repair" through the synergistic effect of fibronectin and mussel protein, solving the pain point of anti-wrinkle ingredients. As an important adhesion molecule, fibronectin can bind to integrin receptors on the surface of skin cells, promote the migration and proliferation of keratinocytes, accelerate the repair of the skin barrier, reduce the secretion of skin irritants, relieve skin redness and stinging, and at the same time help promote collagen synthesis, enhance the firming and anti-wrinkle effect, like an "engineer" of skin repair, actively mobilizing the skin's own repair potential; mussel protein contains lysine and dopa, which can form a microscopic biological scaffold, adsorb skin cells, and quickly adhere to the wall for repair, while forming a physical isolation layer to reduce external stimulation, like a "guardian" of the skin, quickly relieving discomfort. After the two are combined, they can enhance the collagen synthesis-promoting effect of fibronectin, achieve the synergistic effect of "soothing without weakening anti-wrinkle, and anti-wrinkle without aggravating sensitivity", completely solve the technical problem that traditional anti-wrinkle ingredients are easy to irritate sensitive skin, and fill the technical gap of "synergistic enhancement of anti-wrinkle effect by repair proteins".

[0025] (4) In this invention, oat firming protein and silk core protein work synergistically to achieve “gentle nourishment + enhanced efficacy”, improving product compatibility and efficacy sustainability: oat firming protein can be quickly absorbed by the skin, and achieves soothing and anti-inflammatory effects by inhibiting the NF-κB inflammatory pathway and reducing the release of pro-inflammatory factors. At the same time, it promotes the production of transforming growth factor β, thereby promoting fibroblast proliferation and collagen synthesis, and assists in enhancing firming and anti-wrinkle effects. Its low molecular weight peptides and characteristic amino acids can also enhance the skin’s moisturizing ability and reduce the potential irritation of other protein components. Silk core protein can combat fibroblast aging by improving mitochondrial function, inhibiting oxidative stress, blocking the MAPK pathway, inhibiting collagen degradation, and self-assembling to form an ECM-like scaffold, promoting fibroblast proliferation, promoting collagen secretion, enhancing anti-wrinkle and firming effects, and inhibiting inflammatory factors to repair the damaged skin barrier. When combined, the two ingredients significantly improve the gentleness of the entire protein system, while enhancing the synergistic effect of firming, anti-wrinkle, and soothing repair. This avoids the irritation of single protein ingredients, making it suitable for various skin types such as sensitive and aging skin. It also extends the duration of efficacy, addressing the issue of weak efficacy persistence in existing products.

[0026] (5) This invention achieves a synergistic effect of "firming, anti-wrinkle, soothing, and repairing" through the precise compounding of oat firming protein, silk protein, recombinant collagen, recombinant fibronectin, recombinant mussel protein, and Mayinglong Babao powder. Among them, oat firming protein can tighten the skin matrix immediately, silk protein and recombinant collagen work together to fill and support the dermal structure, effectively reducing wrinkles; recombinant fibronectin promotes cell adhesion and migration, accelerates epidermal barrier repair, and recombinant mussel protein forms a protective film with strong adhesion properties, stabilizing the local microenvironment; zinc carbonate hydroxide in Mayinglong Babao powder provides astringent, anti-inflammatory, and soothing effects, while amber powder and pearl powder promote blood circulation and tissue regeneration. Each component targets four dimensions: skin firmness, wrinkle depth, inflammatory response, and barrier damage, forming a cascade complementary network of "mechanical support - cell activation - inflammation regulation - barrier reconstruction", thereby comprehensively improving skin condition. Detailed Implementation

[0027] The technical solutions of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of protection of the present invention.

[0028] Unless otherwise specified, all raw and auxiliary materials used in this invention are readily available and their specifications comply with national pharmaceutical standards.

[0029] As partly exemplary, information on some of the substances used in the embodiments of the present invention is as follows: The Mayinglong Babao raw powder is produced by Mayinglong Health Industry Co., Ltd.; the Mayinglong Babao raw powder used in this embodiment of the invention is obtained by mixing zinc carbonate hydroxide, amber powder and pearl powder in a mass ratio of 98.5:0.5:1. The recombinant collagen is a triple-complex recombinant collagen TTA1BC-01, purchased from Guangzhou Huiju Biotechnology Co., Ltd. The oat-based skin-firming protein was purchased from Beijing Dongfang Miaosen Biotechnology Co., Ltd. The silk protein is silk protein oil sponge-meadowfoam seed oil 40, purchased from Shanghai Xianke Biotechnology Co., Ltd. The recombinant mussel protein is commercially known as MAP-Reno M5E05C and was purchased from Shenzhen Baiyin Biotechnology Co., Ltd. The recombinant fibronectin is marketed as HythermFN-01 and was purchased from Shenzhen Baiyin Biotechnology Co., Ltd.

[0030] Example 1 A cosmetic composition comprising 0.01 parts of Mayinglong Babao raw powder, 3 parts of oat firming protein, 0.1 parts of silk protein, 0.01 parts of recombinant collagen, 0.01 parts of recombinant fibronectin, and 0.001 parts of recombinant mussel protein.

[0031] Example 2 A cosmetic composition comprising 0.005 parts of Mayinglong Babao raw powder, 2.8 parts of oat firming protein, 0.2 parts of silk protein, 0.02 parts of recombinant collagen, 0.005 parts of recombinant fibronectin, and 0.002 parts of recombinant mussel protein.

[0032] Example 3 A cosmetic composition comprising 0.015 parts of Mayinglong Babao raw powder, 3.2 parts of oat firming protein, 0.15 parts of silk protein, 0.015 parts of recombinant collagen, 0.004 parts of recombinant fibronectin, and 0.003 parts of recombinant mussel protein.

[0033] Example 4 An essence lotion, comprising the following components by weight percentage: Table 1. Composition of the essence lotion in Example 4 In this embodiment, a method for preparing the above-mentioned essence lotion is also provided, comprising the following steps: (1) Add purified water, disodium EDTA, sodium hyaluronate, dipotassium glycyrrhizate, 1,3-propanediol and glycerol from phase A into an emulsification tank, stir and heat to 85°C to obtain material A; (2) Add polydimethylsiloxane, Olivem 1000, PEG-100 stearate, OLIWAX® LC, POLYSYNLANE LITE, tocopheryl acetate, shea butter, Mayinglong Babao powder, bisabolol, and sorbitan caprylate from phase B into a pretreatment tank, stir and heat to 85°C to obtain material B; (3) Add material B to material A, and stir homogenously to obtain material C; (4) Add SEPINOV WEO and butanediol from phase C to material C, stir homogenize, and then cool to 45°C to obtain material D; (5) Mix the oat skin firming protein, silk core protein, recombinant collagen, recombinant fibronectin and recombinant mussel protein in phase D and add them to material D. Mix them evenly to obtain material E. (6) Add 1,2-hexanediol and ZEASTAT from phase E to material E, stir homogenously until uniform, and cool to room temperature.

[0034] Example 5 An essence lotion, comprising the following components by weight percentage: Table 2 Composition of the essence lotion in Example 5 In this embodiment, a method for preparing the above-mentioned essence lotion is also provided, comprising the following steps: (1) Add purified water, disodium EDTA, sodium hyaluronate, dipotassium glycyrrhizate, 1,3-propanediol and glycerol from phase A into an emulsification tank, stir and heat to 85°C to obtain material A; (2) Add polydimethylsiloxane, Olivem 1000, PEG-100 stearate, OLIWAX® LC, POLYSYNLANE LITE, tocopheryl acetate, shea butter, Mayinglong Babao powder, bisabolol, and sorbitan caprylate from phase B into a pretreatment tank, stir and heat to 85°C to obtain material B; (3) Add material B to material A, and stir homogenously to obtain material C; (4) Add SEPINOV WEO and butanediol from phase C to material C, stir homogenize, and then cool to 45°C to obtain material D; (5) Mix the oat skin firming protein, silk core protein, recombinant collagen, recombinant fibronectin and recombinant mussel protein in phase D and add them to material D. Mix them evenly to obtain material E. (6) Add 1,2-hexanediol and ZEASTAT from phase E to material E, stir homogenously until uniform, and cool to room temperature.

[0035] Example 6 An essence lotion, comprising the following components by weight percentage: Table 3 Composition of the essence lotion in Example 6 In this embodiment, a method for preparing the above-mentioned essence lotion is also provided, comprising the following steps: (1) Add purified water, disodium EDTA, sodium hyaluronate, dipotassium glycyrrhizate, 1,3-propanediol and glycerol from phase A into an emulsification tank, stir and heat to 85°C to obtain material A; (2) Add polydimethylsiloxane, Olivem 1000, PEG-100 stearate, OLIWAX® LC, POLYSYNLANE LITE, tocopheryl acetate, shea butter, Mayinglong Babao powder, bisabolol, and sorbitan caprylate from phase B into a pretreatment tank, stir and heat to 85°C to obtain material B; (3) Add material B to material A, and stir homogenously to obtain material C; (4) Add SEPINOV WEO and butanediol from phase C to material C, stir homogenize, and then cool to 45°C to obtain material D; (5) Mix the oat skin firming protein, silk core protein, recombinant collagen, recombinant fibronectin and recombinant mussel protein in phase D and add them to material D. Mix them evenly to obtain material E. (6) Add 1,2-hexanediol and ZEASTAT from phase E to material E, stir homogenously until uniform, and cool to room temperature.

[0036] Comparative Example 1 An essence lotion, differing from Example 4, comprises a cosmetic composition consisting of 0.01 parts of Mayinglong Babao raw powder, 3 parts of oat firming protein, 0.1 parts of silk protein, 0.01 parts of recombinant collagen, and 0.001 parts of recombinant mussel protein. All other components are the same, with corresponding adjustments made to the preparation method.

[0037] Comparative Example 2 An essence lotion, differing from Example 4, comprises a cosmetic composition consisting of 0.01 parts of Mayinglong Babao raw powder, 3 parts of oat firming protein, 0.1 parts of silk protein, 0.01 parts of recombinant collagen, and 0.01 parts of recombinant fibronectin. All other components are the same, with corresponding adjustments made to the preparation method.

[0038] Comparative Example 3 An essence lotion, differing from Example 4, comprises a cosmetic composition consisting of 0.01 parts of Mayinglong Babao raw powder, 3 parts of oat firming protein, 0.01 parts of recombinant collagen, 0.01 parts of recombinant fibronectin, and 0.001 parts of recombinant mussel protein. All other components are the same, with corresponding adjustments made to the preparation method.

[0039] Comparative Example 4 An essence lotion, differing from Example 4, comprises a cosmetic composition consisting of 3 parts oat firming protein, 0.1 parts silk protein, and 0.01 parts recombinant collagen. All other components are the same, with the preparation method adjusted accordingly.

[0040] Comparative Example 5 An essence lotion, differing from Example 4, comprises 0.01 parts of Ma Yinglong Babao raw powder, 0.01 parts of recombinant fibronectin, and 0.001 parts of recombinant mussel protein. All other components are the same, with corresponding adjustments made to the preparation method.

[0041] Experimental Example 1 This test example refers to the standard "T / SHRH 034-2021 Cosmetic Soothing Efficacy Test - In Vitro TNF-α Inflammatory Factor Content Determination Lipopolysaccharide Induced Macrophage RAW264.7 Test Method" to determine the soothing efficacy of the products prepared in the examples and comparative examples.

[0042] 1.1 Cytotoxicity Detection Press 1×10 4 Cells were seeded at a density of cells / well into 96-well plates and incubated overnight. The experiment included a zeroing group, a control group, a positive control group, and a sample group. The sample group used the product from Example 4, with eight concentration gradients (1%, 0.5%, 0.25%, 0.125%, 0.0625%, 0.0313%, 0.0156%, 0.0078%), with three replicates for each concentration gradient. The control group used culture medium containing 10% PBS; the positive control group used culture medium containing 10% DMSO; the sample group used culture medium containing the corresponding concentration of the sample; the zeroing group had no cells seeded, only cell culture medium. The OD values ​​of different samples were measured, and cell viability was calculated using the formula: Cell viability = (OD of sample wells - OD of zeroing wells) / (OD of solvent control wells - OD of zeroing wells). 100%.

[0043] Based on the cytotoxicity test results, the average cell viability of the 1% concentration serum was 91.66% (cell survival rate ≥80%), and the 1% (m / V) concentration serum was selected as the safe dose.

[0044] 1.2 TNF-α content detection Press 1×10 5 Cells were seeded at a density of cells / well into 24-well plates and incubated overnight. The experiment included a blank control group (no modeling), a negative control group, a positive control group (100 μg / mL dexamethasone), and a sample group (1% m / V of the example and comparative products). When the cell deposition rate in the 24-well plates reached 40%–60%, 100 μL of 10×LPS working solution was added to each well according to the experimental design. The plates were shaken to mix the drug, and the sample was added simultaneously. The final LPS concentration was 1 μg / mL, with three replicates per group. After loading, the plates were incubated for another 24 h. After incubation, the cell culture supernatant was collected, and the TNF-α inhibition rate was measured according to the Mouse TNF-α ELISA kit instructions. The TNF-α inhibition rate was calculated as follows: TNF-α inhibition rate (%) = (1 - average TNF-α content of the test substance / average TNF-α content of the negative control) × 100%. The test results are shown in Table 4.

[0045] Table 4. TNF-α data of the serums prepared in the comparative examples and embodiments. (Compared with the blank control group, #p<0.05, ##p<0.01; compared with the negative control group, p < 0.05 p < 0.01 As shown in the table, compared with the blank control group, the secretion of the inflammatory factor TNF-α in RAW264.7 macrophages in the negative control group was significantly increased (p<0.01), indicating that LPS stimulation modeling was successful in this experiment. Compared with the negative control group, the secretion of TNF-α in the positive control group was significantly decreased (p<0.01), indicating that the positive control test was effective. Compared with the negative control group, the secretion of TNF-α by the product of the example was significantly reduced (p<0.01), with an inhibition rate of 26.37%, and the inhibition rate of TNF-α by the product of the example was significantly better than that of the comparative product, indicating that the product of the example of this invention can better inhibit the secretion of the inflammatory factor TNF-α in RAW264.7 macrophages, and the soothing effect of the product of the example is better than that of the comparative product.

[0046] Experimental Example 2 This test example refers to the standard "T / SHRH 031-2020 Test of Firming and Anti-wrinkle Efficacy of Cosmetics - Determination of Type I Collagen Content in In Vitro Fibroblasts" to determine the firming and anti-wrinkle efficacy of the products prepared in the examples and comparative examples.

[0047] 2.1 Cytotoxicity Detection Press 2×10 4 HSF cells were seeded into 96-well plates at a seeding density of [number] cells / well and incubated overnight. Five concentration gradients were set up (50, 100, 200, 400, 800, 1600 μg / mL), with three replicates for each gradient. Cytotoxicity was then assessed according to the CCK-8 assay instructions, and cell viability was calculated. Cell viability (%) = (mean absorbance of test sample / mean absorbance of solvent control) × 100%.

[0048] Based on the cytotoxicity test results, the cell survival rate of the 800 μg / mL serum was 92.18% (cell survival rate ≥80%), and 800 μg / mL serum was selected as the safe dose.

[0049] 2.2 Detection of Type I Collagen Content HSF cells were seeded into 96-well plates at a seeding density of 2 × 10⁴ cells / well and incubated overnight. The experiment included a blank control group (cell culture medium), a positive control group (TGF-β1 solution), and a sample group (800 μg / mL of the example and comparative products). After incubation, cell supernatant was collected from each well, and then analyzed according to the corresponding ELISA kit instructions. The upregulation rate of type I collagen content was calculated using the formula: Upregulation rate (%) = ((T-SC) / SC) × 100%; where T is the average type I collagen content of the test samples, and SC is the average type I collagen content of the blank control. The results are shown in Table 5.

[0050] Table 5. Type I collagen content data of products in comparative examples and case studies. (Compared with the blank control group, p < 0.05 p < 0.01 As shown in the table, compared with the blank control group, the type I collagen content in the positive control group was significantly increased (p<0.01), indicating that the positive control test was effective. Compared with the blank control group, the type I collagen content of the product in the example was significantly increased (p<0.01), with an increase rate of 48.37%, and the increase rate of the product in the example was significantly better than that of the comparative product, indicating that the product in the example of the present invention has a better ability to promote the synthesis of type I collagen, and the firming and anti-wrinkle effects of the product in the example are better than those of the comparative product.

[0051] Experimental Example 3 This experimental example uses a cell scratch assay to scratch a cell monolayer and captures images using a time-lapse microscope to determine the repair efficacy of the products prepared in the examples and comparative examples.

[0052] Using a ruler as a guide, draw horizontal lines evenly at the bottom of each 12-well plate with a marker, spaced 0.5 cm apart. The lines should extend through the wells, with at least five lines per well. Harvested HaCaT cells in good growth condition were trypsin-digested to prepare a single-cell suspension, which was then seeded into 12-well plates. DMEM medium containing 10% fetal bovine serum (FBS) was added, and the plates were incubated at 37 °C and 5% CO2 for 24 h until the cells reached full confluence at the bottom of the wells. Using a 200 μL pipette tip attached to a ruler, the wells were marked perpendicularly to the lines. The cells were washed three times with PBS buffer to remove any detached cells. Serum-free DMEM medium was added, and the images were photographed under a microscope as the initial images at 0 h. The experiment included a blank control group, a positive control group (10 ng / mL EGF), and a sample group (0.1% w / v of the example and comparative products). All groups were incubated under the same conditions for another 24 h before being photographed under a microscope.

[0053] Cell migration rate was calculated using the formula: Cell migration rate (%) = (Initial scratch area - 24h scratch area) / Initial scratch area × 100%; the scratch area was measured using ImageJ software. A higher migration rate indicates a better repair effect of the composition. The test results are shown in Table 6.

[0054] Table 6. Cell migration rate data of the serums prepared in the comparative examples and embodiments. As shown in the table, compared with the blank control group, the content of type I collagen in the positive control group was significantly increased, indicating that the positive control test was effective. Compared with the blank control group, the cell migration rate of the product in the example was significantly increased, and the cell migration rate of the product in the example was significantly better than that of the comparative product, indicating that the product in the example of the present invention has better repair efficacy.

[0055] The applicant declares that the present invention is illustrated by the above embodiments, but the present invention is not limited to the above embodiments, that is, it does not mean that the present invention must rely on the above embodiments to be implemented. Those skilled in the art should understand that any improvements to the present invention, equivalent substitutions of individual raw materials in the product of the present invention, addition of auxiliary components, selection of specific methods, etc., all fall within the protection scope and disclosure scope of the present invention.

Claims

1. A multi-protein synergistic firming, anti-wrinkle, soothing, and repairing cosmetic composition, characterized in that it comprises: The product is prepared by weight from the following raw materials: 0.001~0.1 parts of Mayinglong Babao raw powder, 0.01~5 parts of oat skin-firming protein, 0.01~1 parts of silk protein, 0.001~0.1 parts of recombinant collagen, 0.001~0.1 parts of recombinant fibronectin, and 0.0001~0.1 parts of recombinant mussel protein.

2. The multi-protein synergistic firming, anti-wrinkle, soothing, and repairing cosmetic composition according to claim 1, characterized in that, The product is prepared by weight from the following raw materials: 0.005~0.05 parts of Mayinglong Babao raw powder, 0.1~4 parts of oat skin-firming protein, 0.01~0.5 parts of silk protein, 0.005~0.05 parts of recombinant collagen, 0.005~0.05 parts of recombinant fibronectin, and 0.0005~0.05 parts of recombinant mussel protein.

3. The multi-protein synergistic firming, anti-wrinkle, soothing, and repairing cosmetic composition according to claim 1, characterized in that, The product is prepared by weight from the following raw materials: 0.01 parts of Mayinglong Babao raw powder, 3 parts of oat skin-firming protein, 0.1 parts of silk protein, 0.01 parts of recombinant collagen, 0.01 parts of recombinant fibronectin, and 0.001 parts of recombinant mussel protein.

4. The multi-protein synergistic firming, anti-wrinkle, soothing, and repairing cosmetic composition according to claim 1, characterized in that, The product is prepared by weight from the following raw materials: 0.005 parts of Mayinglong Babao raw powder, 2.8 parts of oat skin-firming protein, 0.2 parts of silk protein, 0.02 parts of recombinant collagen, 0.005 parts of recombinant fibronectin, and 0.002 parts of recombinant mussel protein.

5. The multi-protein synergistic firming, anti-wrinkle, soothing, and repairing cosmetic composition according to claim 1, characterized in that, The product was prepared by weight from the following raw materials: 0.015 parts of Mayinglong Babao raw powder, 3.2 parts of oat skin-firming protein, 0.15 parts of silk protein, 0.03 parts of recombinant collagen, 0.004 parts of recombinant fibronectin, and 0.0015 parts of recombinant mussel protein.

6. The use of the cosmetic composition according to any one of claims 1 to 5 in the preparation of cosmetics.

7. An essence comprising, It includes the cosmetic composition and excipients described in any one of claims 1 to 5.

8. The serum of claim 7, wherein the serum is characterized by, The excipients are selected from one or more of the following: humectants, soothing agents, antioxidants, oils, water, chelating agents, emulsifiers, preservatives, polyols, and thickeners.

9. The essence lotion according to claim 8, characterized in that, The moisturizer is sodium hyaluronate; The soothing agent is one or two of dipotassium glycyrrhizate and bisabolol; The polyol is selected from one or more of glycerol, butylene glycol, sorbitol, dipropylene glycol, and 1,3-propanediol; The oil is selected from one or more of the following: polydimethylsiloxane, polydimethylsiloxane alcohol, cyclopentadimethylsiloxane, polydimethylsiloxane cross-linked polymer, hydrogenated polyisobutylene, isopropyl myristate, caprylic / capric triglyceride, isononyl isononanoate, neopentyl glycol dicaprylate / dicaprylate, squalane, and shea butter. The chelating agent is selected from one or two of disodium EDTA and octanoyl hydroxamic acid; The emulsifier is selected from one or more of isocetyl palmitate, cetearyl glucoside, cetearyl alcohol, PEG-100 stearate, cetearyl oleate, and sorbitan oleate; The antioxidant is selected from one or two of tocopherol and tocopherol acetate; The preservative is selected from one or more of phenoxyethanol, ethylhexylglycerin, chlorphenesin, methylparaben, p-hydroxyacetophenone, propylparaben, 1,2-hexanediol, and sorbitan octanoate; The thickener is selected from one or more of the following: polyacrylate crosspolymer-6, hydroxyethyl acrylate / sodium acryloyldimethyl taurate copolymer, sorbitan isostearate, sodium polyacryloyldimethyl taurate, ammonium acryloyldimethyl taurate / VP copolymer, sodium polyacrylate, polyacrylamide, acrylate / C10-30 alkanol acrylate crosspolymer, and carbomer.

10. A method for preparing the essence lotion according to any one of claims 1 to 7, characterized in that, Includes the following steps: (1) Add water, chelating agent, moisturizer, first part soothing agent and first part polyol to emulsification tank, stir and heat to 80-85℃ to obtain material A; (2) Add the oil, emulsifier, antioxidant, Mayinglong Babao powder, second part of the soothing agent, and first part of the preservative into the pretreatment tank, stir and heat to 80-85℃ to obtain material B; (3) Add material B to material A, and stir homogenously to obtain material C; (4) Add the thickener and the second part of the polyol to material C, stir homogenize and cool to 40~45℃ to obtain material D; (5) Mix oat firming protein, silk core protein, recombinant collagen, recombinant fibronectin and recombinant mussel protein and add to material D. Homogenize and stir evenly to obtain material E. (6) Add the second part of the preservative to material E, stir homogenously until uniform, and cool to room temperature.

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