A multi-ceramide-containing repair composition and a method for preparing the same

Abstract

This invention discloses a repair composition containing multiple ceramides and its preparation method, belonging to the field of cosmetic technology. The repair composition, by mass parts, comprises ceramide AP, ceramide EOP, ceramide NP, phytosphingosine, Centella asiatica extract, Portulaca oleracea extract, Hydrangea macrophylla extract, ectoine, and tripeptide-1. This invention utilizes multiple ceramides and phytosphingosine to biomimetically replenish the lipids of the stratum corneum, combined with the synergistic effects of various plant active ingredients and active peptides, to effectively achieve multiple effects such as skin barrier repair, soothing and anti-inflammatory properties, and anti-allergy. This invention also provides a barrier repair serum containing this repair composition and its preparation method. The serum has stable physicochemical properties and, verified by in vitro cell experiments and human efficacy tests, can rapidly repair damaged skin barriers, improve sensitive symptoms such as redness and stinging, and is gentle and non-irritating, suitable for daily skincare use by people with sensitive skin and damaged skin barriers.

Description

Technical Field

[0001] This invention relates to the field of cosmetic technology, and in particular to a repair composition containing multiple ceramides and its preparation method. Background Technology

[0002] The skin barrier is the first line of defense for the human skin, mainly composed of stratum corneum cells and intercellular lipids. Intercellular lipids, primarily composed of ceramides, cholesterol, and free fatty acids, play a crucial role in locking in skin moisture and resisting the invasion of external irritants. With the fast pace of modern life, environmental pollution, over-cleansing, frequent exfoliation, and improper skincare are causing increasing numbers of people to experience damaged skin barriers. Symptoms include dryness, flaking, redness, stinging, itching, and other sensitive symptoms, severely impacting skin health and quality of life.

[0003] Most barrier repair products on the market currently only use a single ceramide ingredient for lipid replenishment or add only a single plant-based soothing ingredient. These products suffer from limited efficacy and poor synergy, making it difficult to simultaneously achieve multiple effects such as barrier repair, soothing and anti-inflammatory properties, and anti-allergy. Some products, in pursuit of rapid results, add irritating ingredients, which can further burden sensitive skin and fail to meet the skincare needs of people with sensitive skin. Therefore, developing a repair composition that can work synergistically across multiple dimensions to simultaneously achieve barrier repair, soothing and anti-inflammatory properties, and anti-allergy, while also being gentle and stable, has significant market demand and application value. Summary of the Invention

[0004] The purpose of this invention is to address the shortcomings of existing technologies by providing a repair composition. This composition, through the scientific combination of various ceramides, phytosphingosine, and various active ingredients, can significantly stimulate the migration of HaCaT cells, while simultaneously exerting soothing, anti-inflammatory, and anti-allergic effects, thus synergistically achieving multiple skin care benefits.

[0005] To achieve the above objectives, the present invention adopts the following technical solution: In a first aspect, the present invention provides a repair composition comprising, by weight parts: 0.1-0.5 parts of ceramide AP, 0.1-0.5 parts of ceramide EOP, 0.1-0.5 parts of ceramide NP, 0.05-0.3 parts of phytosphingosine, 1-3 parts of Centella asiatica extract, 2-4 parts of Portulaca oleracea extract, 0.5-2 parts of Hydrangea macrophylla extract, 0.5-2 parts of ectoine, and 0.01-0.1 parts of tripeptide-1.

[0006] Preferably, the repair composition comprises, by weight, the following components: 0.3 parts of ceramide AP, 0.2 parts of ceramide EOP, 0.4 parts of ceramide NP, 0.15 parts of phytosphingosine, 2 parts of Centella asiatica extract, 3 parts of Portulaca oleracea extract, 1.2 parts of Hydrangea macrophylla extract, 1.2 parts of ectoine, and 0.05 parts of tripeptide-1.

[0007] Preferably, the repair composition is prepared by heating ceramide AP, ceramide EOP, ceramide NP and phytosphingosine to 70-80°C and stirring until completely melted, cooling to 40-50°C and then adding the remaining components in sequence, stirring evenly and then cooling to room temperature, and storing in a sealed container away from light.

[0008] The design principle of the composition of this invention lies in the following: Ceramides AP, EOP, and NP are key components of intercellular lipids in skin cells. A specific ratio of these compounds more closely resembles the natural lipid composition of the human body, effectively repairing the "brick wall structure." Phytosphingosine is a precursor to ceramides and can promote the skin's own synthesis of ceramides. Centella asiatica extract (rich in asiaticoside) and Portulaca oleracea extract (rich in polysaccharides and alkaloids) exert synergistic anti-inflammatory and soothing effects, reducing skin irritation. Hydrangea macrophylla extract is rich in β-glucan, which can form a protective film on the skin surface and activate skin immunity. Ectoin is a highly effective cell protectant that can stabilize proteins and biomembranes, enhancing the skin's tolerance to environmental stress. Tripeptide-1 (GHK) is a signal peptide that can promote collagen and glycosaminoglycan synthesis, supporting barrier repair from the dermis. The components work synergistically through multiple pathways—"lipid replenishment, soothing and anti-inflammatory, external protection, and internal repair"—to achieve comprehensive barrier repair.

[0009] Secondly, the present invention provides the use of the above-described repair composition in the preparation of skincare products with barrier repair, soothing, and / or anti-allergic effects. The skincare product dosage forms include, but are not limited to, creams, lotions, serums, and masks.

[0010] Thirdly, the present invention provides a barrier repair serum containing 1-5 wt% of the above-mentioned repair composition.

[0011] Furthermore, the essence also includes 3-8 wt% of a moisturizer, 0.1-1 wt% of a thickener, 0.1-0.5 wt% of a preservative, and 0.1-0.5 wt% of an emulsifier, with the balance being deionized water. The moisturizer may be selected from one or more of glycerin, panthenol, sodium hyaluronate, and β-glucan; the thickener may be selected from one or more of carbomer, xanthan gum, and sclerotium gum; the preservative may be selected from one or more of succinate, potassium sorbate, and sodium benzoate; and the emulsifier may be selected from one or more of PEG-40 hydrogenated castor oil, polysorbate-80, and sucrose fatty acid esters.

[0012] Fourthly, the present invention provides a method for preparing the above-mentioned barrier repair essence, comprising the following steps: S1: Heat deionized water to 75-85℃, add thickener, stir evenly to obtain aqueous phase A; S2: Cool the aqueous phase A to 40-50℃, add the humectant, emulsifier and preservative in sequence, stir evenly to obtain the aqueous phase B; S3: Cool the aqueous phase B to 35-45℃, add the repair composition, and stir until uniform to obtain the barrier repair essence.

[0013] The beneficial effects of this invention are as follows: The repair composition provided by this invention scientifically combines specific types and proportions of ceramides, their biosynthetic precursors, multi-source plant active ingredients, and cell-protective factors to construct a multi-target, multi-pathway synergistic repair system. This system is not a simple superposition of the functions of its components, but rather achieves systemic and comprehensive repair of the skin barrier through the synergistic and enhanced effects of the following intrinsic mechanisms: Biomimetic replenishment and active regeneration work synergistically to efficiently repair the physical barrier: By compounding ceramides AP, EOP, and NP, which have a composition and ratio similar to the intercellular lipids of the stratum corneum in human skin, it can directly and precisely replenish key lipid components in the "brick wall structure," quickly repairing barrier vulnerabilities. Simultaneously, the addition of its biosynthetic precursor, phytosphingosine, promotes the synthesis of ceramides by keratinocytes themselves. This not only achieves "external replenishment" but also initiates an active repair mechanism of "internal synthesis." The synergistic effect of both ensures a long-term, stable supply and repair of barrier lipids.

[0014] The synergistic anti-inflammatory and soothing pathways effectively improve the microenvironment of a damaged skin barrier: Damaged skin barriers are often accompanied by inflammatory responses. In this invention, the combination of Centella asiatica extract and Portulaca oleracea extract can exert a synergistic anti-inflammatory effect by inhibiting the release of key pro-inflammatory factors, thereby reducing the inflammatory cascade response caused by barrier damage at its source. Simultaneously, the active substances contained in both extracts work together to soothe irritation and reduce nerve sensitivity, creating a stable and low-irritation internal skin environment for barrier repair.

[0015] External protection and internal strengthening work synergistically to enhance barrier resistance: The *Hydrangea macrophylla* extract in the composition is rich in β-glucan, which can form a skin-friendly protective film on the skin surface, instantly reducing the invasion of external irritants and the loss of internal moisture, providing physical external protection. Ectoin, as a highly effective cell protectant, can stabilize skin cell membranes and protein structures, improve the cell's tolerance to various environmental stresses, and strengthen barrier function at the cellular level. The signal peptide tripeptide-1 can promote the synthesis of extracellular matrix such as collagen and glycosaminoglycans in the dermis, providing a solid underlying support for the health of the epidermal barrier, achieving "inside-out" strengthening.

[0016] In summary, the composition of this invention repairs the physical structure through "direct biomimetic lipid replenishment" and "promoting self-regeneration of biological precursors," improves the repair microenvironment through "inhibiting the source of inflammation" and "soothing nerve sensations," and enhances barrier resistance and stability through "external film-forming protection," "enhancing cell tolerance," and "strengthening the dermal matrix." The components and functional pathways are interconnected and mutually reinforcing, ultimately achieving a comprehensive, highly effective, and long-lasting multi-functional effect of skin barrier repair, soothing inflammation, and anti-allergy that cannot be achieved with a single ingredient or simple compounding. This effectively overcomes the technical problems of existing products, such as "single efficacy and poor synergy." Detailed Implementation

[0017] To make the objectives, technical solutions, and advantages of the present invention clearer, the embodiments of the present invention will be described in detail below. The following embodiments are used to illustrate the present invention, but are not intended to limit the scope of the invention.

[0018] The sources of some of the raw materials used in this invention are as follows: Ceramide AP is from Guangzhou Xufan Trading Co., Ltd., and the manufacturer is Chongqing Zhihe Biomedical Co., Ltd., with the trade name: Ceramide 6; The ceramide EOP comes from Guangzhou Xufan Trading Co., Ltd., and the manufacturer is Chongqing Zhihe Biomedical Co., Ltd. Ceramide NP was purchased from Guangzhou Zhongran Biotechnology Co., Ltd., product name: Ceramide 3.

[0019] Phytosphingosine comes from Guangzhou Xufan Trading Co., Ltd., and is manufactured by Chongqing Zhihe Biomedical Co., Ltd. Trade name: Phytosphingosine. Centella asiatica extract was purchased from Guangzhou Hengtao Trading Co., Ltd., product name: Centella asiatica extract.

[0020] Purslane extract was purchased from Guangzhou Baihaobo Co., Ltd., product name: Purslane Extract.

[0021] The *Hydrangea macrophylla* extract was purchased from Guangzhou Novogene Biotechnology Co., Ltd., product name: *Hydrangea macrophylla* extract XNY5000.

[0022] Ectocin was purchased from Suzhou Nakang Biotechnology Co., Ltd., product name: Ectocin.

[0023] Tripeptide-1 is from Guangzhou Fuzherui Trading Co., Ltd., and the manufacturer is Guangzhou Yuerui Biotechnology Co., Ltd. The product name is Tripeptide-1 (2000ppm).

[0024] All other raw materials, reagents, and equipment were commercially available.

[0025] Preparation of Repair Composition Composition 1 Composed of the following parts by weight: Ceramide AP: 0.3 parts; Ceramide EOP: 0.2 parts; Ceramide NP: 0.4 parts; Phytosphingosine: 0.15 parts; Centella Asiatica extract: 2 parts; Purslane extract: 3 parts; Hydrangea macrophylla extract: 1.2 parts; Ectocin: 1.2 parts; Tripeptide-1: 0.05 parts; Preparation method: Mix ceramide AP, ceramide EOP, ceramide NP and phytosphingosine evenly and heat to 75°C, stir well, cool to 45°C, then add Centella asiatica extract, Portulaca oleracea extract, Hydrangea macrophylla extract, Ectoin and tripeptide-1 in sequence, stir until even, cool to room temperature, seal and store away from light.

[0026] Composition 2 Composed of the following parts by weight: Ceramide AP: 0.1 parts; Ceramide EOP: 0.1 parts; Ceramide NP: 0.1 parts; Phytosphingosine: 0.05 parts; Centella Asiatica extract: 1 part; Purslane extract: 2 parts; Hydrangea extract: 0.5 parts; Ectocin: 0.5 parts; Tripeptide-1: 0.01 parts; The preparation method is the same as that of composition 1.

[0027] Composition 3 Composed of the following parts by weight: Ceramide AP: 0.5 parts; Ceramide EOP: 0.5 parts; Ceramide NP: 0.5 parts; Phytosphingosine: 0.3 parts; Centella Asiatica extract: 3 parts; Purslane extract: 4 parts; Hydrangea extract: 2 parts; Ectocin: 2 portions; Tripeptide-1: 0.1 parts; The preparation method is the same as that of composition 1.

[0028] Composition A: Unlike Composition 1, it lacks ceramide AP; the missing mass parts are made up by ceramide EOP, ceramide NP and phytosphingosine in a mass ratio of 0.2:0.4:0.15, and the mass parts of the other components are the same as those of Composition 1. Preparation method: Mix ceramide EOP, ceramide NP, and phytosphingosine evenly and heat to 75°C, stirring until homogeneous. Cool to 45°C, then add Centella asiatica extract, Portulaca oleracea extract, Hydrangea macrophylla extract, Ectocin, and tripeptide-1 in sequence, stirring until homogeneous. Cool to room temperature, seal, and store away from light.

[0029] Composition B: Unlike Composition 1, it lacks ceramide EOP; the missing mass parts are made up by ceramide AP, ceramide NP and phytosphingosine in a mass ratio of 0.3:0.4:0.15, and the mass parts of the other components are the same as those of Composition 1; Preparation method: Mix ceramide AP, ceramide NP, and phytosphingosine evenly and heat to 75°C, stirring until homogeneous. Cool to 45°C, then add Centella asiatica extract, Portulaca oleracea extract, Hydrangea macrophylla extract, Ectocin, and tripeptide-1 in sequence, stirring until homogeneous. Cool to room temperature, seal, and store away from light.

[0030] Composition C: Unlike composition 1, it lacks ceramide NP; the missing mass parts are made up by ceramide AP, ceramide EOP and phytosphingosine in a mass ratio of 0.3:0.2:0.15, and the mass parts of the other components are the same as those of composition 1. Preparation method: Mix ceramide AP, ceramide EOP, and phytosphingosine evenly and heat to 75°C, stirring until homogeneous. Cool to 45°C, then add Centella asiatica extract, Portulaca oleracea extract, Hydrangea macrophylla extract, Ectoin, and tripeptide-1 in sequence, stirring until homogeneous. Cool to room temperature, seal, and store away from light.

[0031] Composition D: Unlike Composition 1, it lacks phytosphingosine; the missing mass parts are made up by ceramide AP, ceramide EOP, and ceramide NP in a mass ratio of 0.3:0.2:0.4, and the mass parts of the other components are the same as those of Composition 1; Preparation method: Mix ceramide AP, ceramide EOP, and ceramide NP evenly and heat to 75°C, stirring until homogeneous. Cool to 45°C, then add Centella asiatica extract, Portulaca oleracea extract, Hydrangea macrophylla extract, Ectocin, and tripeptide-1 in sequence, stirring until homogeneous. Cool to room temperature, seal, and store in the dark.

[0032] Composition E Composed of the following components by mass: Ceramide AP: 0.05 parts; Ceramide EOP: 0.4 parts; Ceramide NP: 0.2 parts; Phytosphingosine: 0.4 parts; Centella Asiatica extract: 2 parts; Purslane extract: 3 parts; Hydrangea macrophylla extract: 1.2 parts; Ectocin: 1.2 parts; Tripeptide-1: 0.05 parts; The preparation method is the same as that of composition 1.

[0033] Composition ①: Unlike composition 1, it lacks Centella asiatica extract, and the missing mass parts are made up by Portulaca oleracea extract and Hydrangea spp. extract in a mass ratio of 3:1.2. The mass parts of the other components are the same as those of composition 1. Preparation method: Mix ceramide AP, ceramide EOP, ceramide NP and phytosphingosine evenly and heat to 75°C, stir until uniform, cool to 45°C, add purslane extract, physalis extract, ectoine and tripeptide-1 in sequence, stir until uniform, cool to room temperature, seal and store away from light.

[0034] Composition ②: Unlike composition 1, it lacks purslane extract. The missing mass parts are made up by Centella asiatica extract and Hydrangea macrophylla extract in a mass ratio of 2:1.2. The mass parts of the remaining components are the same as those of composition 1. Preparation method: Mix ceramide AP, ceramide EOP, ceramide NP and phytosphingosine evenly and heat to 75°C, stir until uniform, cool to 45°C, add Centella asiatica extract, Hydrangea macrophylla extract, ectoine and tripeptide-1 in sequence, stir until uniform, cool to room temperature, seal and store away from light.

[0035] Composition ③: Unlike composition 1, it lacks Hydrangea macrophylla extract. The missing mass is made up by Centella asiatica extract and Portulaca oleracea extract in a mass ratio of 2:3. The mass of the remaining components is the same as that of composition 1. Preparation method: Mix ceramide AP, ceramide EOP, ceramide NP and phytosphingosine evenly and heat to 75°C, stir until uniform, cool to 45°C, then add Centella asiatica extract, Portulaca oleracea extract, ectoine and tripeptide-1 in sequence, stir until uniform, cool to room temperature, seal and store away from light.

[0036] Composition ④: Composed of the following parts by weight: Ceramide AP: 0.3 parts; Ceramide EOP: 0.2 parts; Ceramide NP: 0.4 parts; Phytosphingosine: 0.15 parts; Centella asiatica extract: 0.5 parts; Purslane extract: 4 parts; Hydrangea extract: 1.7 parts; Ectocin: 1.2 parts; Tripeptide-1: 0.05 parts; The preparation method is the same as that of composition 1.

[0037] Composition α: Unlike composition 1, it lacks ectoine, and the missing mass fraction is made up with tripeptide-1, while the mass fractions of the other components are the same as those of composition 1; Preparation method: Mix ceramide AP, ceramide EOP, ceramide NP and phytosphingosine evenly and heat to 75°C, stir until uniform, cool to 45°C, then add Centella asiatica extract, Portulaca oleracea extract, Hydrangea macrophylla extract and tripeptide-1 in sequence, stir until uniform, cool to room temperature, seal and store away from light.

[0038] Composition β: Unlike composition 1, it lacks tripeptide-1, and the missing mass fraction is made up with ectoine, while the remaining mass fractions are the same as those in composition 1; Preparation method: Mix ceramide AP, ceramide EOP, ceramide NP and phytosphingosine evenly and heat to 75°C, stir until uniform, cool to 45°C, add Centella asiatica extract, Portulaca oleracea extract, Hydrangea macrophylla extract and ectoine in sequence, stir until uniform, cool to room temperature, seal and store away from light.

[0039] Composition λ Composed of the following parts by weight: Ceramide AP: 0.3 parts; Ceramide EOP: 0.2 parts; Ceramide NP: 0.4 parts; Phytosphingosine: 0.15 parts; Centella Asiatica extract: 2 parts; Purslane extract: 3 parts; Hydrangea macrophylla extract: 1.2 parts; Ectocin: 0.05 parts; Tripeptide-1: 1.2 parts; The preparation method is the same as that of composition 1.

[0040] Composition I: Unlike Composition 1, it lacks ceramide AP, ceramide EOP, ceramide NP, and phytosphingosine; the missing mass parts are made up by Centella asiatica extract, Portulaca oleracea extract, Hydrangea macrophylla extract, ectoine, and tripeptide-1 in a mass ratio of 2:3:1.2:1.2:0.05, and the mass parts of the other components are the same as those of Composition 1. Preparation method: Add Centella asiatica extract, Portulaca oleracea extract, Hydrangea macrophylla extract, Ectocin, and Tripeptide-1 in sequence, stir until homogeneous, seal and store away from light.

[0041] Composition II: Unlike Composition 1, it lacks Centella asiatica extract, Portulaca oleracea extract, and Hydrangea macrophylla extract; the missing mass parts are made up by ceramide AP, ceramide EOP, ceramide NP, phytosphingosine, ectoine, and tripeptide-1 in a mass ratio of 0.3:0.2:0.4:0.15:1.2:0.05, and the mass parts of the other components are the same as those of Composition 1; Preparation method: Mix ceramide AP, ceramide EOP, ceramide NP and phytosphingosine evenly and heat to 75°C, stir until uniform, cool to 45°C, add ectoine and tripeptide-1 in sequence, stir until uniform, cool to room temperature, seal and store away from light.

[0042] Composition III: Unlike Composition 1, it lacks ectoine and tripeptide-1; the missing mass parts are made up by ceramide AP, ceramide EOP, ceramide NP, phytosphingosine, centella asiatica extract, purslane extract, and physalis extract in a mass ratio of 0.3:0.2:0.4:0.15:2:3:1.2, and the mass parts of the other components are the same as those of Composition 1; Preparation method: Mix ceramide AP, ceramide EOP, ceramide NP and phytosphingosine evenly and heat to 75°C, stir until uniform, cool to 45°C, add Centella asiatica extract, Portulaca oleracea extract and Hydrangea macrophylla extract in sequence, stir until uniform, cool to room temperature, seal and store away from light.

[0043] Preparation of Barrier Repair Serum Serum 1 Composed of the following components by mass percentage: Composition 1: 2wt% Moisturizer: 4wt% Thickener: 0.3 wt%; Preservative: 0.5 wt%; Emulsifier: 0.3 wt%; Deionized water to 100 wt%; S1: Heat deionized water to 80°C, add thickener, stir well to obtain aqueous phase A; S2: Cool aqueous phase A to 45°C, add humectant, emulsifier and preservative in sequence, stir evenly to obtain aqueous phase B; S3: Cool the aqueous phase B to 40°C, add the repair composition, and stir until uniform to obtain the barrier repair essence; The humectant is a mixture of glycerin and sodium hyaluronate in a mass ratio of 1:0.4; the thickener is carbomer; the preservative is acetone; and the emulsifier is polysorbate-80.

[0044] Serum 2 Composed of the following components by mass percentage: Composition 1: 1 wt% Moisturizer: 4wt% Thickener: 0.3 wt%; Preservative: 0.5 wt%; Emulsifier: 0.3 wt%; Deionized water to 100 wt%; S1: Heat deionized water to 80°C, add thickener, stir well to obtain aqueous phase A; S2: Cool aqueous phase A to 45°C, add humectant, emulsifier and preservative in sequence, stir evenly to obtain aqueous phase B; S3: Cool the aqueous phase B to 40°C, add the repair composition, and stir until uniform to obtain the barrier repair essence; The humectant is a mixture of glycerin and sodium hyaluronate in a mass ratio of 1:0.4; the thickener is carbomer; the preservative is acetone; and the emulsifier is polysorbate-80.

[0045] Serum 3 Composed of the following components by mass percentage: Composition 1: 5wt% Moisturizer: 4wt% Thickener: 0.3 wt%; Preservative: 0.5 wt%; Emulsifier: 0.3 wt%; Deionized water to 100 wt%; S1: Heat deionized water to 80°C, add thickener, stir well to obtain aqueous phase A; S2: Cool aqueous phase A to 45°C, add humectant, emulsifier and preservative in sequence, stir evenly to obtain aqueous phase B; S3: Cool the aqueous phase B to 40°C, add the repair composition, and stir until uniform to obtain the barrier repair essence; The humectant is a mixture of glycerin and sodium hyaluronate in a mass ratio of 1:0.4; the thickener is carbomer; the preservative is acetone; and the emulsifier is polysorbate-80.

[0046] Blank Essence Composed of the following components by mass percentage: Moisturizer: 4wt% Thickener: 0.3 wt%; Preservative: 0.5 wt%; Emulsifier: 0.3 wt%; Deionized water to 100 wt%; S1: Heat deionized water to 80°C, add thickener, stir well to obtain aqueous phase A; S2: Cool aqueous phase A to 45°C, add humectant, emulsifier and preservative in sequence, stir evenly to obtain aqueous phase B; S3: Cool the aqueous phase B to 40°C and stir until homogeneous to obtain the blank essence solution; The humectant is a mixture of glycerin and sodium hyaluronate in a mass ratio of 1:0.4; the thickener is carbomer; the preservative is acetone; and the emulsifier is polysorbate-80.

[0047] Test Example 1: Effect of the Composition on Repair Efficacy The effects of the various repair compositions of the present invention on the migration ability of HaCaT cells (human immortalized keratinocytes) were evaluated in vitro to simulate their efficacy in promoting skin barrier repair.

[0048] Cell line: HaCaT cells.

[0049] Main reagents: DMEM high glucose medium, fetal bovine serum, penicillin-streptomycin antibiotics, and trypsin.

[0050] Test substances: Compositions of the present invention 1, 2, 3, A, B, C, D, E, ①, ②, ③, ④, α, β, λ, I, II, III. Prepared as a 100 mg / mL stock solution with DMSO, and diluted to the working concentration with serum-free culture medium during the test.

[0051] Positive control: Recombinant human epidermal growth factor solution.

[0052] Main instruments: CO2 incubator, inverted microscope, cell culture plate, cell scratching tool.

[0053] Cell Culture and Plating: HaCaT cells were cultured in DMEM medium containing 10% v / v fetal bovine serum at 37°C in a 5% CO2 incubator. Logarithmic growth phase cells were harvested, digested, counted, and plated at 5 × 10⁻⁶ cells / cells. 4Cells were seeded at a density of 90% per well in 24-well plates and cultured overnight until the cell monolayer confluence reached over 90%.

[0054] Cell scratching: Create a straight scratch in the center of each well using a 200 μL sterile pipette tip. Gently wash twice with PBS to remove detached cells.

[0055] Treatment: The culture medium was replaced with serum-free medium containing different test substances. Grouping was as follows: Blank control group: serum-free culture medium without the composition.

[0056] Positive control group: serum-free culture medium containing 10 ng / mLEGF.

[0057] Experimental group: serum-free culture medium containing each composition (to achieve a final concentration of 10 μg / mL in the culture system).

[0058] Observation and photography: At 0h and 24h after the scratch, the same scratch area was photographed under an inverted microscope (100×).

[0059] Cell migration rate (%) = [(0h scratch area - 24h scratch area) / 0h scratch area] × 100%.

[0060] Table 1. Effect of the composition on cell migration rate (mean ± SD, n=3) Group Cell migration rate (%) Blank control group 18.5±3.2 Positive control group 63.7±4.1 Composition 1 57.3±3.8 Composition 2 53.6±3.5 Composition 3 55.1±3.9 Composition A 40.2±2.9# Composition B 41.8±3.1# Composition C 39.7±2.7# Composition D 43.5±3.0# Composition E 36.4±2.8# Composition ① 46.8±3.3# Composition ② 47.1±3.0# Composition ③ 52.6±3.5 Composition ④ 44.9±3.1# Composition α 55.1±3.7 Composition β 53.9±3.6 Composition λ 54.1±3.7 Composition I 32.8±2.5# Composition II 38.9±2.8# Composition III 54.8±3.4 Note: Compared with composition 1, "#" P<0.05.

[0061] The results showed that the preferred compositions 1, 2, and 3 of this invention significantly promoted the migration of HaCaT cells, indicating their excellent potential for promoting epidermal repair. When any one of the ceramides (A, B, C) or the precursor phytosphingosine (D) was missing, the migration-promoting effect decreased statistically significantly, confirming the synergistic necessity of specific types and ratios of ceramides and precursors for mimicking intercellular lipids and promoting barrier repair. Composition E also showed a decreased effect due to an imbalance in the ratio of ceramides to phytosphingosine. The migration-promoting effects of compositions ③ (lacking *Hydrangea macrophylla* extract), α (lacking ectoine), β (lacking tripeptide-1), and III (lacking both ectoine and tripeptide-1) were not significantly different from those of composition 1, indicating that the contributions of *Hydrangea macrophylla* extract, ectoine, and tripeptide-1 were relatively minor in the in vitro cell migration model. The effects of compositions ① and ② (lacking *Centella asiatica* or *Portulaca oleracea* extract) were significantly lower than those of composition 1, indicating that these two extracts made a positive contribution to promoting repair. Compositions I and II (lacking ceramide and phytosphingosine components and plant extract components, respectively) showed the weakest effects, demonstrating that the absence of core lipid-replenishing or anti-inflammatory soothing components significantly reduces the overall repair effect. In summary, this invention, through the scientific formulation of multiple components, demonstrates a significant synergistic effect in promoting cell migration and mimicking barrier repair.

[0062] Test Example 2: Effect of the Composition on TNF-α Secretion The anti-inflammatory efficacy of each composition was evaluated by detecting its effect on the secretion of tumor necrosis factor-α by lipopolysaccharide-induced macrophages.

[0063] Cell line: mouse monocyte-macrophage RAW264.7.

[0064] Main reagents: DMEM culture medium, LPS, TNF-α ELISA detection kit.

[0065] Test substances: Compositions 1, 2, 3, A, B, C, D, E, ①, ②, ③, ④, α, β, λ, I, II, III of the present invention. Prepared as a 100 mg / mL stock solution with DMSO, and diluted to the working concentration with serum-free culture medium during the experiment; the working concentration was determined to be non-cytotoxic through preliminary experiments.

[0066] Main instruments: CO2 incubator, ELISA reader.

[0067] Cell culture and seeding: RAW264.7 cells were cultured in DMEM containing 10% FBS. The cell density was adjusted to 2 × 10⁻⁶ cells / year. 5 Cells / mL were seeded into 96-well plates, 100 μL per well, and incubated overnight.

[0068] Drug administration and induction: Discard the old culture medium and then replenish it with the same volume of test sample as the old culture medium. The test samples are grouped as follows: Normal control group: serum-free culture medium without LPS and test substance.

[0069] Model control group: serum-free culture medium containing 1 μg / mL LPS.

[0070] Experimental group: serum-free culture medium containing 1 μg / mL LPS and each test substance (final concentration 20 μg / mL).

[0071] Incubation and detection: After culturing for another 24 hours, the cell supernatant was collected and the concentration of TNF-α in the supernatant was determined according to the instructions of the TNF-α ELISA kit.

[0072] Inhibition rate (%) = [1 - (TNF-α concentration in experimental group - TNF-α concentration in normal control group) / (TNF-α concentration in model control group - TNF-α concentration in normal control group)] × 100% The data for each group are shown in the table below.

[0073] Table 2. Effect of the composition on the inhibition rate of TNF-α secretion. Group TNF-α inhibition rate (%) Composition 1 68.6 Composition 2 63.2 Composition 3 65.9 Composition A 56.7 Composition B 57.6 Composition C 53.1 Composition D 59.1 Composition E 49.4 Composition ① 58.1 Composition ② 57.7 Composition ③ 55.0 Composition ④ 55.3 Composition α 66.2 Composition β 65.8 Composition λ 64.1 Composition I 33.9 Composition II 42.5 Composition III 55.3 Data show that compositions 1, 2, and 3 within the scope defined in this invention all exhibit an inhibition rate of over 60% on TNF-α secretion, demonstrating significant anti-inflammatory activity. Among them, composition 1, with the preferred formulation, shows the highest inhibition rate, reaching 68.6%.

[0074] Compared to composition 1, composition II (lacking Centella asiatica extract, Portulaca oleracea extract, and Hydrangea macrophylla extract) showed a sharp decrease in anti-inflammatory inhibition rate, strongly confirming that these three plant extracts are the core components for the anti-inflammatory effect of this invention, and their synergistic effect is crucial for inhibiting the key pro-inflammatory factor TNF-α. The inhibition rate of composition I (lacking all ceramides and phytosphingosine) was also significantly reduced to 33.9%, indicating that a complete skin lipid replenishment system plays a positive role in maintaining cell stability and indirectly assisting in anti-inflammation. The inhibition rate of composition III (lacking ectoine and tripeptide-1) was also significantly lower than that of composition 1, indicating that these two components have a clear enhancing effect on the anti-inflammatory effect of this invention. The effects of compositions A, B, C, D, and E were all lower than those of composition 1 to varying degrees, indicating that the combination of specific types and proportions of ceramides and phytosphingosine is a necessary condition for achieving the best synergistic anti-inflammatory effect. The effects of compositions α, β, λ, and III were similar to or slightly lower than those of composition 1, indicating that in this anti-inflammatory model, minor adjustments to the ratio of these two components did not significantly affect the final effect. The effects of compositions ①-④ were also lower than those of composition 1, demonstrating that the specific ratio of the three extracts has a synergistic effect.

[0075] In summary, this invention achieves a significant synergistic effect in inhibiting the key pro-inflammatory factor TNF-α by combining specific types and proportions of ceramides, phytosphingosine, and extracts of Centella asiatica, Portulaca oleracea, and Hydrangea macrophylla. The absence of any core component or an imbalance in its proportion will lead to a decrease in anti-inflammatory effect.

[0076] Test Example 3: Inhibition of Hyaluronidase Activity by the Composition The anti-allergy potential of each composition was evaluated by measuring its inhibitory effect on hyaluronidase activity.

[0077] Solvent preparation: Aqueous solution of polysorbate-80, wherein the concentration of polysorbate-80 is 0.3 wt%.

[0078] Test samples: Compositions 1, 2, 3, A, B, C, D, E, ①, ②, ③, ④, α, β, λ, I, II, III of this invention. Prepare a 100 mg / mL stock solution with a solvent, and dilute with distilled water to the working concentration during the test; sonicate for 10 minutes until completely dissolved during the preparation of the stock solution, and the working concentration was determined to be non-cytotoxic through preliminary experiments.

[0079] (1) Take four clean test tubes and label them A, B, C and D respectively. Add 50 μL of the sample to be tested with a concentration of 10 mg / L to each of test tubes A and B. Add 50 μL of distilled water to each of test tubes C and D. Add 50 μL of 500 U / mL hyaluronidase solution to each of test tubes A and C. Add 50 μL of acetate buffer solution with pH = 5.6 to each of test tubes B and D. Place the test tubes in a 37℃ incubator for 20 min.

[0080] (2) Add 10 μL of 2.5 mol / L CaCl2 solution to each of the four test tubes, and then place them in a 37℃ incubator for 20 min.

[0081] (3) Add 50 μL of 0.5 mg / mL sodium hyaluronate solution to test tubes A and C, and add 50 μL of pH 5.6 acetate buffer to test tubes B and D. Place the test tubes in a 37℃ incubator for 40 min, and then take them out and let them stand at room temperature for 10 min.

[0082] (4) Add 50 μL of distilled water, 10 μL of 5 mol / L NaOH solution and 50 μL of acetylacetone solution to the four test tubes respectively. Place the test tubes in a water bath for 30 min of boiling water, then in an ice bath for 10 min, and finally in a room temperature bath for 10 min.

[0083] (5) Add 100 μL of P-DAB reagent to each test tube, and then measure the absorbance at 530 nm using an ELISA reader.

[0084] Hyaluronidase inhibition rate (%) = [1 - (AB) / (CD)] × 100% In the formula: A is the OD value of the sample solution; B is the OD value of the blank sample; C is the OD value of the control sample solution; and D is the OD value of the blank control sample.

[0085] Table 3. Effect of the composition on hyaluronidase inhibition rate Group Hyaluronidase inhibition rate (%) Composition 1 78.6±4.1 Composition 2 74.4±3.8 Composition 3 75.9±3.9 Composition A 65.3±3.2# Composition B 61.8±3.0# Composition C 63.7±3.1# Composition D 68.2±3.4# Composition E 58.9±2.9# Composition ① 65.3±3.2# Composition ② 61.8±3.0# Composition ③ 70.5±3.5 Composition ④ 66.4±3.3# Composition α 76.1±3.9 Composition β 77.0±4.1 Composition λ 75.3±3.8 Composition I 40.5±2.9# Composition II 35.2±2.5# Composition III 69.8±3.6# Note: Compared with composition 1, "#" P<0.05.

[0086] The data clearly show that compositions 1, 2, and 3 all exhibit strong inhibitory activity against hyaluronidase (inhibition rate >72%), confirming that the compositions of the present invention have direct anti-allergic potential.

[0087] The inhibition rate of Composition II (lacking all plant extracts) decreased significantly to 35.2%, and the inhibition rate of Composition I (lacking all ceramides and phytosphingosine) also decreased significantly to 40.5%, both far lower than that of Composition 1. This demonstrates that Centella asiatica extract, Portulaca oleracea extract, and Hydrangea macrophylla extract are the core active components that exert direct anti-allergic (hyaluronidase inhibition) effects, while the complete ceramide repair system also plays an important role in maintaining skin barrier integrity and indirectly assisting in anti-allergy; both are indispensable. When any of the three ceramides (AP, EOP, NP) or their precursor phytosphingosine is missing (Compositions A, B, C, D) or their ratio is imbalanced (Composition E), the inhibition rate shows a statistically significant decrease. Similarly, an imbalance in the ratio of the three plant extracts (Composition ④) also leads to a decrease in efficacy. The inhibition rates of Compositions α, β, λ (adjusting the ratio of ectoine to tripeptide-1) and Composition III (lacking both) are not significantly different from those of Composition 1, indicating that in this hyaluronidase inhibition experiment, ectoine and tripeptide-1 are not the key components that exert direct anti-allergic effects.

[0088] Test Example 4: Security Test The biocompatibility of the composition was verified by experiments in this invention.

[0089] The irritation of cosmetics was evaluated using the 2015 Cosmetic Safety Technical Specifications as a reference standard. The test method was a skin patch test. Subjects aged 18-60 years were tested and randomly assigned to all test products, with each subject corresponding to all test products, for a total of 30 subjects.

[0090] Test substances: Compositions 1, 2, 3, A, B, C, D, E, ①, ②, ③, ④, α, β, λ, I, II, III of the present invention. Prepare a 10wt% test substance with distilled water, shake vigorously before use, and use after thorough mixing.

[0091] Test Method: Eighteen test areas were divided on the subject's back. The test substance was placed in a patch applicator at a dosage of 0.020-0.025g. The patch applicator containing the test substance was then covered with non-irritating adhesive tape and applied to the test area on the subject's back. Gently press with the palm of the hand to ensure even adhesion to the skin surface. Each test area corresponded to one test substance. The treatment lasted for 24 hours. Thirty minutes after removing the patch applicator, and after the pressure marks disappeared, the skin reaction was observed. If the result was negative, observations were repeated at 24 and 48 hours post-pattern testing.

[0092] Evaluation criteria: Grade 0: Negative reaction; Grade 1: Suspicious reaction, with only slight erythema; Grade 2: Weak positive reaction, erythema, infiltration, edema, and possible papules; Grade 3: Strong positive reaction, erythema, infiltration, edema, papules may be present, and the reaction may extend beyond the test area; Grade 4: Extremely positive reaction, with obvious erythema, severe infiltration, edema, confluent herpes, and reaction extending beyond the test area.

[0093] Test results: All subjects showed negative skin reactions, indicating that the test substance provided by this invention is safe and non-irritating.

[0094] Test Example 5: Effect of Composition on TEWL Value Human trials were conducted to evaluate the efficacy of serums containing the compositions of this invention in improving skin barrier function (indicated by TEWL values).

[0095] Subject selection: Female volunteers who meet the inclusion criteria (age 25-45 years, dry or sensitive skin, healthy facial skin) are recruited. Subjects are required to be grouped according to the test sample, with each group corresponding to one test sample, and each group has 5 people.

[0096] Test area: face.

[0097] Test samples: Compositions of the present invention 1, 2, 3, A, B, C, D, E, ①, ②, ③, ④, α, β, λ, I, II, III. Prepare test samples with a concentration of 2 wt% using deionized water; the blank test sample is deionized water, which should be vigorously shaken and mixed before use.

[0098] Product Application: Apply the test sample to the test area. Dosage: 1 mg / cm² per area. 2 Once a day.

[0099] Measurement indicators: The TEWL (g / h·m³) values ​​of each area were measured using a Tewameter®™ 300 transdermal moisture loss meter before use (0h) and after 28 days of continuous use. 2 ).

[0100] Data analysis: Calculate the rate of change of TEWL values ​​and perform statistical analysis.

[0101] TEWL reduction rate (%) = (TEWL value before use (0h) - TEWL value after 28 days of use) / TEWL value before use (0h) × 100% Table 4. Effect of the composition on the reduction rate of TEWL Group TEWL reduction rate (%) Composition 1 28.5±1.2 Composition 2 25.1±2.9 Composition 3 27.8±3.0 Composition A 20.3±2.5# Composition B 21.0±2.6# Composition C 19.8±2.4# Composition D 22.5±2.7# Composition E 18.2±2.3# Composition ① 23.6±2.8# Composition ② 22.1±2.7# Composition ③ 23.1±2.9# Composition ④ 21.8±2.8# Composition α 23.3±3.1# Composition β 21.8±3.0# Composition λ 24.1±2.8# Composition I 12.1±2.0# Composition II 15.8±1.9# Composition III 23.8±2.7# Blank sample 2.5±1.5# Note: Compared with composition 1, "#" P<0.05.

[0102] Data shows that compositions 1, 2, and 3 within the scope of this invention can significantly reduce the skin's TEWL value after 28 days of continuous use. Among them, composition 1, with its preferred formulation, shows the best effect. The significant reduction in TEWL value directly proves that the skin barrier function is effectively enhanced and moisture loss is reduced.

[0103] Compositions A, B, C, and D (each lacking one ceramide or phytosphingosine) and composition E all showed significantly lower barrier repair effects than composition 1. Composition I had the weakest effect, which fully demonstrates that the combination of specific types and proportions of ceramides and phytosphingosine is the foundation and core for achieving physical barrier repair. The barrier repair effect of composition II was also significantly reduced. This indicates that the anti-inflammatory and soothing effects of Centella asiatica and Portulaca oleracea extracts are crucial for alleviating the inflammatory state associated with barrier damage and creating a skin microenvironment conducive to barrier repair. Composition ③, lacking Hydrangea macrophylla extract, showed no significant difference in effect from composition 1, possibly because it primarily provides immediate external protection, while its contribution to improving TEWL within 28 days is relatively indirect. The effects of compositions α, β, and λ were significantly lower than those of composition 1, indicating that ectoine and tripeptide-1 make a positive contribution to the consolidation of barrier function by promoting cell tolerance and matrix synthesis.

[0104] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit the scope of protection of the present invention. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the essence and scope of the technical solutions of the present invention.

Claims

1. A repair composition, characterized in that, The composition comprises the following components in parts by weight: Ceramide AP: 0.1-0.5 parts; Ceramide EOP: 0.1-0.5 parts; Ceramide NP: 0.1-0.5 parts; Phytosphingosine: 0.05-0.3 parts; Centella Asiatica extract: 1-3 parts; Purslane extract: 2-4 parts; Hydrangea extract: 0.5-2 parts; Ectocin: 0.5-2 parts; Tripeptide-1: 0.01-0.1 parts.

2. The repair composition according to claim 1, characterized in that, The composition comprises the following components in parts by weight: Ceramide AP: 0.3 parts; Ceramide EOP: 0.2 parts; Ceramide NP: 0.4 parts; Phytosphingosine: 0.15 parts; Centella Asiatica extract: 2 parts; Purslane extract: 3 parts; Hydrangea macrophylla extract: 1.2 parts; Ectocin: 1.2 parts; Tripeptide-1: 0.05 parts.

3. The use of a repair composition according to claim 1 or 2 in the preparation of skin care products having barrier repair, soothing and / or anti-allergic effects.

4. The application according to claim 3, characterized in that, The skincare product formulation is any one of cream, lotion, serum, or mask.

5. A barrier repair essence, characterized in that, Contains 1-5 wt% of the repair composition as described in claim 1 or 2.

6. The barrier repair essence according to claim 5, characterized in that, It also includes 3-8 wt% humectant, 0.1-1 wt% thickener, 0.1-0.5 wt% preservative, 0.1-0.5 wt% emulsifier, and the balance is deionized water.

7. The barrier repair essence according to claim 6, characterized in that, The humectant is one or more of glycerin, panthenol, sodium hyaluronate, and β-glucan; the thickener is one or more of carbomer, xanthan gum, and sclerotium gum; the preservative is one or more of succinate, potassium sorbate, and sodium benzoate; and the emulsifier is one or more of PEG-40 hydrogenated castor oil, polysorbate-80, and sucrose fatty acid ester.

8. A method for preparing a barrier repair essence according to claim 6 or 7, characterized in that, Includes the following steps: S1: Heat deionized water to 75-85℃, add thickener, stir evenly to obtain aqueous phase A; S2: Cool the aqueous phase A to 40-50℃, add the humectant, emulsifier and preservative in sequence, stir evenly to obtain the aqueous phase B; S3: Cool the aqueous phase B to 35-45℃, add the repair composition, and stir until uniform to obtain the barrier repair essence.