Composition with basement membrane repairing effect as well as preparation method and application of composition

Abstract

The invention discloses a composition with a basilar membrane repairing effect and preparation and application of the composition, and the composition is prepared from the following components in percentage by mass: 0.01%-0.2% of recombinant IV type collagen, 0.4%-4.0% of yeast fermentation product extract, 0.01%-0.2% of acetyl tetrapeptide-9, 0.01%-0.2% of acetyl tetrapeptide-11, 4%-10% of 1, 2-hexanediol, 0.01%-0.5% of ethylhexylglycerin and the balance of water. The composition has the advantages that the skin cell activity can be greatly improved, and cell migration is promoted; and the repair of the basement membrane is effectively protected and promoted through the ways of remarkably promoting the growth of the components of the basement membrane, enhancing the skin barrier, resisting inflammation, relieving and the like.

Description

Technical Field

[0001] This invention relates to the field of cosmetic technology, and more particularly to cosmetics that repair the basement membrane. Background Technology

[0002] The skin basement membrane, located between epithelial cells and stromal cells in animals, is a soft yet resilient extracellular matrix structure with a thickness of 50–100 nm. It separates the basal cells of the epidermis from the underlying connective tissue, provides structural support to the cells, maintains epidermal-dermal intercellular function through regulation by epidermal-dermal cytokines, and also serves as a certain permeability barrier.

[0003] The main components of the skin's basement membrane are: type IV collagen (COLⅣ) and type VII collagen (COLⅦ), laminin (LN), nidogen / entactin, perlecan, etc. COLⅣ and laminin are present in the stratum compactum, forming a network structure that is crucial for the stability of the basement membrane. Laminin, proteoglycans, and heparin sulfate are present in the stratum lucidum. COLⅦ pierces the stratum lucidum and binds to hemidesmosomes in the stratum compactum; COLⅦ is present in the sub-stratum compactum. Nidogen / entactin and perlecan bridge the laminin and COLⅣ network structure, enhancing the stability of the basement membrane and influencing its structural integrity.

[0004] The skin's basement membrane, with type IV collagen at its core and combined with laminin and other components, plays a multi-dimensional role in skincare. It physically blocks harmful external substances through precise pore size, reducing the loss of moisture and nutrients from the dermis and forming a deep barrier; it filters small-molecule nutrients to supply the epidermis and regulates epidermal renewal homeostasis by binding to cell receptors; it strengthens the epidermal-dermal connection through an anchoring system, preventing skin delamination and sagging, and also regulates dermal collagen synthesis for anti-aging; when damaged, it releases signaling peptides to guide cells to rebuild the basement membrane, regulate inflammation, accelerate healing, and reduce pigmented scars.

[0005] Basement membrane damage is caused by a combination of exogenous stimuli and endogenous factors. Among exogenous factors, ultraviolet radiation is the primary cause, activating enzymes to degrade type IV collagen. Mechanical friction, improper micro-invasive procedures, extreme temperatures, and irritating skincare products can also directly or indirectly damage its structure. Endogenous factors are related to natural aging (decreased collagen synthesis and increased degradation), inflammatory immune abnormalities (inflammatory factors accelerate damage), and nutritional metabolic disorders (deficiency of key nutrients or metabolic diseases affecting supply). After damage, the skin often appears sensitive and red, with visible blood vessels, fine lines, and uneven pigmentation. Functionally, it manifests as barrier decline leading to dry skin, decreased repair capacity, and susceptibility to microbial infections. Therefore, protecting the integrity of the basement membrane can fundamentally improve skin problems such as sensitivity, dryness, and aging.

[0006] While there are various skincare and cosmetic products on the market, many irritate the skin due to their ingredients and have limited repair effects on the basement membrane. Therefore, developing a gentle, soothing product that effectively repairs the basement membrane and addresses related skin problems at their root would have enormous potential applications. Summary of the Invention

[0007] The purpose of this invention is to provide a composition for basement membrane repair, its preparation method, and its application, in order to solve the problems in the prior art such as the lack of gentle, soothing, and effective products for repairing the basement membrane and improving skin condition.

[0008] To achieve the above objectives, the present invention adopts the following technical solution:

[0009] A composition for basement membrane repair, comprising, by weight percentage: 0.01%–0.2% recombinant type IV collagen, 0.4%–4.0% yeast fermentation product extract, 0.01%–0.2% acetyl tetrapeptide-9, 0.01%–0.2% acetyl tetrapeptide-11, 4%–10% 1,2-hexanediol, 0.01%–0.5% ethylhexylglycerin, with the balance being water.

[0010] Furthermore, the recombinant type IV collagen was prepared from the engineered strain Pichia pastoris GS115-pPIC9K-COL4A1, which was obtained from the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 30004 and is disclosed in patent application CN 118852407 A.

[0011] Furthermore, the yeast fermentation product extract was prepared by Pichia pastoris fermentation.

[0012] The present invention also provides a method for preparing the composition for basement membrane repair, the preparation process of which is as follows:

[0013] 1) Weigh out the yeast fermentation product extract, recombinant type IV collagen, acetyl tetrapeptide-9, and acetyl tetrapeptide-11 as needed, add them to water, and stir to dissolve them completely to obtain the aqueous phase;

[0014] 2) Weigh 1,2-hexanediol and ethylhexylglycerol, mix them evenly to obtain the alcohol phase, then mix the alcohol phase with the above aqueous phase, and make up to the specified weight with purified water, stirring and mixing evenly;

[0015] 3) Finally, filter and sterilize the solution obtained in the previous step to obtain the composition.

[0016] The present invention also provides the application of the composition for basement membrane repair in the preparation of cosmetics.

[0017] Furthermore, the dosage form of the cosmetic is any one of aqueous solution, lotion, cream, or serum.

[0018] The advantages of this invention are: the composition is water-soluble, making it suitable for developing cosmetic products in various dosage forms. It not only significantly enhances skin cell vitality and promotes cell migration, but also effectively protects and promotes basement membrane repair through mechanisms such as significantly promoting the production of basement membrane components, strengthening the skin barrier, and providing anti-inflammatory and soothing effects. Specific characteristics and advantages are as follows:

[0019] 1) Synergistic repair, covering the entire basement membrane repair process: Recombinant type IV collagen directly replenishes the basement membrane structural framework, acetyl tetrapeptide-9 enhances adhesion, acetyl tetrapeptide-11 promotes the synthesis and secretion of basement membrane components, and yeast fermentation product extract protects the basement membrane microenvironment; the above components work synergistically to form a closed loop of "structural reconstruction - adhesion enhancement - signal activation - inflammation protection", comprehensively covering all aspects of basement membrane repair.

[0020] 2) Scientific and precise skincare: Acetyl tetrapeptide-9 and acetyl tetrapeptide-11 are targeted active peptides that can precisely promote the synthesis of key structural proteins of the basement membrane such as lumican and COLXVII. Compared with skincare ingredients with broad effects, this combination is more precise and efficient, and can specifically solve skin aging-related problems.

[0021] 3) Gentle and non-irritating, suitable for various skin types: Yeast fermentation product extract is derived from natural fermentation and has low skin irritation; Collagen is an inherent component of human skin and is highly safe for external supplementation; Acetyl tetrapeptide-9 and acetyl tetrapeptide-11 are small molecule peptides, which are stable and gentle and generally do not cause adverse reactions such as allergies, making them suitable for most skin types, including sensitive skin.

[0022] In addition, the composition has a simple formulation, high safety and significant efficacy, and can be directly applied to the development of a variety of cosmetic products. Attached Figure Description

[0023] The accompanying drawings, which are included to provide a further understanding of the invention and form part of this application, do not constitute an undue limitation of the invention. In the drawings:

[0024] Figure 1 This is a statistical chart of the results of cytotoxicity testing experiments;

[0025] Figure 2-3 This is a statistical graph showing the gene level detection results of the composition promoting the production of basement membrane constituent proteins;

[0026] Figure 4-5This is a statistical chart showing the test results of the composition indirectly protecting the basement membrane function by enhancing the skin barrier;

[0027] Figure 6 This is a statistical graph showing the detection and resolution of the composition's enhancement of basement membrane function through cellular behavior.

[0028] Figure 7 This is a statistical chart of the results of the cell migration assay;

[0029] Figure 8-9 This is a statistical chart of the results of a cell proliferation experiment. Detailed Implementation

[0030] The present invention will now be described in detail with reference to the accompanying drawings and specific embodiments. The illustrative embodiments and descriptions of the present invention are used to explain the present invention, but are not intended to limit the present invention.

[0031] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. Where specific experimental steps or conditions are not specified in the embodiments, they can be performed according to the conventional experimental procedures or conditions described in the literature in this field. Reagents or instruments whose manufacturers are not specified are all commercially available conventional reagent products.

[0032] 1. Examples 1-3, Comparative Examples

[0033] This specification provides Examples 1-3 and comparative compositions. The components of each composition, by mass percentage, are shown in Table 1.

[0034] Table 1: Examples and Comparative Compositions (mass percentage %)

[0035] The compositions of Examples 1-3 were prepared according to the following steps:

[0036] 1) Weigh out the yeast fermentation product extract, recombinant type IV collagen, acetyl tetrapeptide-9 and acetyl tetrapeptide-11 as needed and add them to purified water. Stir to dissolve completely to obtain the aqueous phase.

[0037] 2) Weigh 1,2-hexanediol and ethylhexylglycerol, mix them evenly to obtain the alcohol phase, then mix the alcohol phase with the above aqueous phase, and make up to the specified weight with purified water, stirring and mixing evenly;

[0038] 3) Finally, filter and sterilize the solution obtained in the previous step to obtain the composition.

[0039] The comparative compositions were prepared according to the preparation process of Examples 1-3, and the corresponding steps were omitted for components that were not present.

[0040] 2. Cytotoxicity testing of Examples 1-3 and comparative compositions

[0041] Cytotoxicity testing can assess whether the ingredients in a product composition will cause damage to cells, such as inhibiting proliferation or inducing apoptosis, thus avoiding skin irritation, allergies, and other problems caused by the ingredients. It can screen for safe raw materials, providing a basis for cosmetic research and development, while also complying with regulatory requirements, controlling product safety from the source, and ultimately protecting consumers' skin health and reducing the occurrence of adverse events.

[0042] Experimental method: Human fibroblasts (HFB) were used at 1× Cells were seeded per well in a 96-well plate. After 24 h of culture, the compositions of Examples 1-3 and the comparative examples were diluted with culture medium at two gradients of 2.5% and 1% and added to the cells in each group. A control group (SC, cells + culture medium) and a blank group (BC, culture medium) were also established. After 24 h of culture, 200 μL of 0.5 mg / mL MTT solution was added to each well. After 4 h, the supernatant was removed, and 150 μL of DMSO was added. The mixture was shaken thoroughly, and after complete color development, the absorbance of each well was measured at 490 nm using a microplate reader. The relative cell viability was calculated using the following formula:

[0043]

[0044] Relative cell viability (%) = (Average absorbance at each concentration ÷ Average absorbance of the control group) × 100%

[0045] The results are as follows Figure 1 As shown in the cytotoxicity test, the relative cell viability of the compositions of Examples 1-3 was ≥80% when diluted to the test concentrations of 2.5% and 1%, indicating that the compositions of Examples 1-3 had no significant cytotoxicity to HFB cells and had reliable safety.

[0046] 3. Detection of gene levels that promote the production of basement membrane constituent proteins by the composition.

[0047] The structural stability and normal function of the basement membrane depend on the synergistic action of multiple proteins. These proteins can be divided into two main categories: collagen family and non-collagenous glycoproteins, which respectively undertake core functions such as structural support, cell anchoring, and signal regulation, and together maintain the integrity of the epidermal-dermal junction (DEJ) of the skin.

[0048] Type IV collagen (COLⅣ) self-assembles into a three-dimensional network framework, providing the structural basis for the basement membrane. Type VII collagen (COLⅦ) forms anchor fibrils, connecting the basement membrane to the dermis; Type XVII collagen (COLⅩⅦ), as a transmembrane collagen, anchors the basal cells of the epidermis to the basement membrane.

[0049] Laminin helps cells bind to the basement membrane and guides epidermal cell differentiation; Nidogen acts as a molecular glue, connecting laminin and type IV collagen to strengthen structural integrity; Perlecan adsorbs moisture and growth factors to maintain microenvironment stability.

[0050] The expression levels of genes such as COLⅣ, COLⅦ, COLⅩⅦ, Laminin, Nidogen, and Perlecan in cells can reflect the basement membrane repair efficacy.

[0051] Experimental methods: Real-time quantitative PCR was used to ionize human fibroblasts (HFB) in the logarithmic growth phase at a concentration of 6× Inoculate 6-well plates at a seeding density of 10 cells / well and incubate at 37 ℃ (5%). Incubate overnight in the solution. When the cell deposition rate in the 6-well plates reaches 40%–50%, divide the cells into the following groups: blank group (BC), control group, and sample group (Examples 1-3 and comparative examples). The drug administration for each group is as follows:

[0052] Add 2 mL of culture medium to the blank group;

[0053] The control group was treated with 2 mL of TGF-β1, with a final concentration of 100 ng / mL, and served as a positive control.

[0054] For the sample group, add 2 mL of the corresponding sample at a concentration of 1% to each well.

[0055] After each group has completed drug administration, the 6-well plates are placed in an incubator (37 ℃, 5%). Cells were incubated in PBS for 24 h. After incubation, the cells were washed three times with 2 mL / well PBS, and 1 mL RNAiso Plus was added to each well. After cell lysis by pipetting, the cell lysates were collected for RNA extraction, reverse transcription, and quantitative real-time PCR to detect the gene levels of COLⅣ, COLⅦ, COLXVII, Laminin, Nidogen, and Perlecan. The method is used to calculate the results.

[0056] Experimental results are as follows Figure 2 , Figure 3 As shown: At a 1% effective concentration, the gene expression levels of COLⅣ, COLⅦ, COLXVII, Laminin, Nidogen, and Perlecan in the sample groups of Examples 1-3 all showed varying degrees of increase compared with the comparative sample group and the blank control group, indicating that the composition can promote the production of type IV collagen, type VII collagen, type XVII collagen, laminin, nestin, and taenin, and has a good repair effect on the basement membrane.

[0057] 4. Detection of the composition's indirect protection of the basement membrane function by enhancing the skin barrier.

[0058] Aquaporin 3 (AQP3) is a key carrier of epidermal water circulation, directly affecting the water content of the stratum corneum and serving as an upstream guarantee for the barrier's water-locking ability. Filamentin (FLG) is a core component of the keratinized capsule (CE, the outer armor of keratinocytes) and the sole source of natural moisturizing factor (NMF), possessing both structural reinforcement and active moisturizing functions. Lorheological protein (LOR) is a key component of CE, building a permeability barrier in the stratum corneum to reduce the penetration of harmful substances (such as bacteria and chemical irritants) into the deeper layers of the epidermis, preventing direct damage to the basement membrane structure. Keratin 1 (K1) is the core of the cytoskeleton of the stratum spinosum-granular layer of the epidermis. It forms a heterodimer with keratin 10 (K10), providing mechanical support for upper epidermal cells and forming the basis for the barrier's resistance to friction and damage. Tight junction protein 4 (CLDN4) is a core component of epidermal tight junctions (TJs, located between cells in the granular layer), serving as the upstream defense line of the barrier and directly controlling the transcellular transport of water and harmful substances. Inner lamina protein (IVL) is a key functional protein in the differentiation of epidermal keratinocytes. It does not directly participate in the construction of the basement membrane, but it strengthens the mechanical stability, impermeability, and moisture retention of the stratum corneum and helps maintain the order of epidermal differentiation. It provides key support for the basement membrane to isolate external damage, stabilize the physiological microenvironment, and ensure anchoring function by strengthening the mechanical stability, impermeability, and moisture retention of the stratum corneum and helping to maintain the order of epidermal differentiation.

[0059] Although AQP3, FLG, LOR, K1, CLDN4, and IVL are not direct structural components of the basement membrane, they all indirectly safeguard the three core functions of the basement membrane—physical barrier, cell anchoring, and signal regulation—by maintaining the stability of the epidermal layer structure, strengthening the epidermal barrier, and reducing factors that damage the basement membrane. They work synergistically with the basement membrane through pathways that enhance the epidermal barrier, reduce damage, and optimize the microenvironment; abnormalities in any of these proteins can indirectly lead to basement membrane dysfunction.

[0060] By detecting the expression of the genes for these proteins in cells, changes in the function of the skin's basement membrane can be reflected.

[0061] Experimental methods: Real-time quantitative PCR was used to analyze human keratinocytes (HaCaT) in the logarithmic growth phase at a concentration of 6 × 10⁻⁶ cells / year. 5 Inoculate 6-well plates at a seeding density of 10 cells / well and incubate at 37 ℃ (5%). Incubate overnight in the solution. When the cell deposition rate in the 6-well plate reaches 40%–50%, divide the cells into the following groups: blank group, control group, and sample group (Examples 1-3 and comparative examples). The drug administration for each group is as follows:

[0062] Add 2 mL of culture medium to the blank group;

[0063] The control group was treated with 2 mL of WY14643 at a final concentration of 50 μM, serving as a positive control.

[0064] Sample group, 2 mL per well, 1% concentration.

[0065] After drug administration, place the 6-well plate in an incubator (37 ℃, 5%). Cells were incubated in PBS for 24 h. After incubation, the cells were washed three times with 2 mL / well PBS, and 1 mL RNAiso Plus was added to each well. After cell lysis by pipetting, the cell lysates were collected for RNA extraction, reverse transcription, and quantitative real-time PCR to detect the gene levels of AQP3, FLG, LOR, K1, CLDN4, and IVL. The method is used to calculate the results.

[0066] Experimental results are as follows Figure 4 , Figure 5 As shown: at a 1% effective concentration, the expression levels of AQP3, FLG, LOR, K1, CLDN4, and IVL genes in the sample groups of Examples 1-3 all showed varying degrees of increase compared with the blank group and the control group, indicating that the composition can protect the normal physiological function of the basement membrane from three aspects: maintaining the stability of the epidermal structure, strengthening the epidermal barrier, and reducing basement membrane damage.

[0067] 5. Testing of the composition's anti-inflammatory, soothing, and protective effects on the basement membrane.

[0068] Interleukin-1α (IL-1α), interleukin-1β (IL-1β), and tumor necrosis factor-α (TNF-α) are early initiators and core amplifiers of skin inflammatory responses. They damage the three core functions of the basement membrane—physical barrier, cell anchoring, and signal regulation—from different dimensions by degrading the basement membrane structure, inhibiting repair synthesis, and disrupting cell adhesion. They often form a synergistic effect that exacerbates the damage.

[0069] By detecting the expression of the above-mentioned pro-inflammatory factors in cells, the impact of inflammation on the structure and function of the skin's basement membrane can be reflected.

[0070] Real-time quantitative PCR was used. RAW264.7 cells in logarithmic growth phase were divided into groups of 6.4 × 10⁻⁶ cells. 5 Seeds were inoculated into 6-well plates at a density of 100 cells / well and incubated in an incubator (37 ℃, 5%). The cells were incubated overnight in PBS. Experimental groups included: blank group (BC), model group (NC), positive control group (dexamethasone 20 μM), and sample group (Examples 1-3 and comparative examples). The blank group was added with fresh culture medium, the model group (NC) with culture medium containing 0.2 μg / mL LPS, and the sample group with culture medium containing 0.2 μg / mL LPS and the sample (1% concentration). After 24 h of culture, the cells were washed twice with 2 mL / well PBS, and 1 mL of RNAiso Plus was added to each well. After cell lysis by pipetting, the cell lysate was collected for RNA extraction, reverse transcription, and quantitative real-time PCR to detect the expression levels of soothing-related genes. The method is used to calculate the results.

[0071] Experimental results are as follows Figure 6 As shown, at a 1% effective concentration, the gene expression levels of IL-1α, IL-1β, and TNF-α in the sample groups of Examples 1-3 were all reduced to varying degrees compared with the model group and the control group, indicating that the composition has an anti-inflammatory and soothing effect, thereby reducing the damage of inflammation to the basement membrane.

[0072] 6. Detection of the composition's enhancement of basement membrane function through cellular behavior

[0073] High-fiber stem cells (HFFBs) are the primary producers of structural components of the basement membrane. Their proliferation and migration directly determine the structural integrity and repair capacity of the basement membrane, thus affecting all its core functions. In normal skin, HFFBs maintain a low rate of proliferation, continuously synthesizing basement membrane components to replenish natural metabolic losses, ensuring stable basement membrane thickness and a dense structure. HFB migration mainly occurs in tissue damage or remodeling scenarios, directly impacting the scope and efficiency of basement membrane repair.

[0074] HaCaT cells are epidermal cells in direct contact with the basement membrane. Their proliferation and migration not only depend on the adhesion and support of the basement membrane, but also conversely regulate the renewal and repair of the basement membrane. HaCaT proliferation mainly occurs in the basal layer of the epidermis (the area in direct contact with the basement membrane), and its proliferation rate is adapted to the adhesion capacity of the basement membrane. HaCaT migration is a core step in the repair of epidermal defects. Its migration process depends entirely on the scaffolding effect of the basement membrane, and it also promotes the functional recovery of the basement membrane.

[0075] The effects of the compositions from Examples 1-3 and the comparative examples on the proliferation and migration of HaCaT and HFB cells can reflect changes in basement membrane function. The following experiments were performed on HaCaT and HFB cells respectively:

[0076] Cell migration assay: Cells in the logarithmic growth phase were divided into groups of 3 × 10⁻⁶ cells. 5 Inoculate 6-well plates at a density of 10 cells / well and incubate at 37°C (5%). Incubate overnight in a medium. When the cell coat rate reaches 70% or higher, perform a scratch assay. Use a 10 µL pipette tip to make two vertical scratches in each well of a six-well plate, using the pipette tip perpendicular to the edge of the ruler, maintaining a 2 cm spacing between the scratches. After making the vertical scratches, make horizontal scratches near the center axis of the six-well plate, perpendicular to the baseline. Wash cells three times with PBS after scratching. Experimental groups are as follows: blank group (BC), positive control group, and sample group (Examples 1-3 and comparative examples). The blank group received 2 mL of serum-free culture medium, the positive control group received 50 μM WY14643 (HaCaT) and 100 ng / mL TGF-β1 (HFB), and the sample group received 1.0% drug concentration, with 2 mL per well. Incubate at 37 ℃ (5%). Continue culturing for 24 hours. Take photos after the scratch and during the culturing process: six photos were taken consecutively at 0 h and 24 h under a 4x microscope. Statistical results are as follows: Figure 7 As shown.

[0077] Cell proliferation assay: MTT assay was used to detect cell proliferation at 5× [a specific concentration]. Cells were seeded at a concentration of 100 μL / well in 96-well plates and cultured for 24 h to detect cell viability. Experimental groups were: blank group (BC, culture medium), control group (SC, cells + culture medium), and sample groups (Examples 1-3 and comparative examples). The concentration of the sample group was 1.0%, with three replicate wells. Other drug administration and culture methods were the same as in the cytotoxicity experiment. Cell viability was measured, and proliferation curves were plotted based on cell viability. Results are as follows: Figure 8 , Figure 9 As shown.

[0078] The experimental results above show that, at a 1% effective concentration, the cell proliferation rate of the sample groups in Examples 1-3 was significantly higher than that of the control group (SC) and the comparative group; similarly, the cell migration rate of the sample groups in Examples 1-3 was significantly higher than that of the blank group (BC) and the comparative group. These results indicate that the compositions of Examples 1-3 promote cell proliferation and migration, and can promote the structural repair and functional recovery of the basement membrane.

[0079] 7. Human safety testing of the composition

[0080] The procedure was performed with reference to the 2015 edition of the "Cosmetic Safety Technical Specifications": the closed skin patch test method in human skin patch testing.

[0081] Select an area not exceeding 50 mm 2A qualified spot treatment device with a depth of approximately 1 mm was used. The four compositions from Examples 1-3 and the comparative example were placed in the small chamber of the spot treatment device, with a volume of 0.020 mL-0.025 mL (liquid). The spot treatment device containing the composition was applied to the flexor side of the forearm of the subjects (33 subjects corresponding to each composition) using hypoallergenic adhesive tape. The device was gently pressed with the palm of the hand to ensure even application to the skin, and left for 24 hours. Skin reactions were observed 30 min (after the indentation disappeared), 24 h, and 48 h after removing the spot treatment device, and the results were recorded.

[0082] The results are shown in Table 2:

[0083] Table 2. Results of closed skin patch tests of the compositions of Examples 1-3 and Comparative Examples.

[0084] The results of the closed skin patch test in Table 1 show that the compositions prepared in Examples 1-3 and the comparative examples of the present invention have good safety and no adverse reactions in humans.

[0085] The technical solutions provided by the embodiments of the present invention have been described in detail above. Specific examples have been used to illustrate the principles and implementation methods of the embodiments of the present invention. The descriptions of the embodiments above are only for helping to understand the principles of the embodiments of the present invention. At the same time, for those skilled in the art, there will be changes in the specific implementation methods and application scope based on the embodiments of the present invention. Therefore, the content of this specification should not be construed as a limitation of the present invention.

Claims

1. A composition for basement membrane repair, characterized in that, By weight percentage, the components include: 0.01%–0.2% recombinant type IV collagen, 0.4%–4.0% yeast fermentation product extract, 0.01%–0.2% acetyl tetrapeptide-9, 0.01%–0.2% acetyl tetrapeptide-11, 4%–10% 1,2-hexanediol, 0.01%–0.5% ethylhexylglycerin, and the balance being water.

2. The composition for basement membrane repair according to claim 1, characterized in that, The recombinant type IV collagen was prepared from Pichia pastoris engineered strain GS115-pPIC9K-COL4A1.

3. The composition for basement membrane repair according to claim 1, characterized in that, The yeast fermentation product extract was prepared by fermentation of Pichia pastoris.

4. A method for preparing a composition for basement membrane repair as described in any one of claims 1-3, characterized in that, The preparation process is as follows: 1) Weigh out the yeast fermentation product extract, recombinant type IV collagen, acetyl tetrapeptide-9, and acetyl tetrapeptide-11 as needed, add them to water, and stir to dissolve them completely to obtain the aqueous phase; 2) Weigh 1,2-hexanediol and ethylhexylglycerol, mix them evenly to obtain the alcohol phase, then mix the alcohol phase with the above aqueous phase, and make up to the specified weight with purified water, stirring and mixing evenly; 3) Finally, filter and sterilize the solution obtained in the previous step to obtain the composition.

5. The use of a composition with the basement membrane repair function as described in any one of claims 1-3 in the preparation of cosmetics.

6. The application according to claim 5, characterized in that, The cosmetic product can be any one of the following formulations: aqueous solution, lotion, cream, or serum.

Images