Peptide complexes and their applications

Abstract

The present invention provides a polypeptide complex and its application, which belongs to the technical field of medical or cosmetic preparations. The present invention synergistically matches three specific polypeptides to obtain a polypeptide complex with the effect of targeted repair of the basement membrane of the skin. The polypeptide complex has the advantages of small molecular weight, high safety, low allergic risk, and strong permeability, and has excellent repair and anti-aging functions. Experiments have shown that the polypeptide complex provided by the present invention can significantly promote the production of endogenous IV, VII, and XVII collagen in the skin as an active peptide raw material, and is also beneficial for repairing the extracellular matrix of the skin, resisting DNA damage, improving skin cyclins, inhibiting the release of inflammatory factors, etc. It can effectively repair the basement membrane and has broad application prospects.

Description

Technical Field

[0001] The present invention relates to the technical field of medical or cosmetic preparations, in particular to a polypeptide complex and applications thereof. Background Art

[0002] One of the core manifestations of skin aging is the structural collapse of the epidermal-dermal junction (DEJ), which is mainly manifested by the degradation of the type IV, type VII, and type XVII collagen networks. As the dynamic interface between the epidermis and dermis, the DEJ not only undertakes the physiological functions of mechanical anchoring and nutrient exchange, but also serves as a core hub for maintaining the integrity of the skin barrier and regulating cell signal transduction. The integrity of its three-dimensional network structure depends on the three-dimensional synergy of the basement membrane skeleton composed of type IV collagen, the anchoring fiber system formed by type VII collagen, and the transmembrane signal transduction network mediated by type XVII collagen. With age, the DEJ layer will become significantly thinner, and the loss of reticular ridges leads to a reduction in its surface area, resulting in a flat appearance, which in turn affects the skin's resistance to shear force and makes the skin more susceptible to damage.

[0003] Existing anti-aging solutions mainly focus on exogenous macromolecular collagen supplementation or single peptide targeting, but these methods have obvious limitations. For example, exogenous macromolecular collagen supplementation has difficulty effectively penetrating the DEJ layer and rebuilding its complex three-dimensional network structure; while single peptide targeting has limited targets and cannot fully activate the key signaling pathways of the DEJ layer. In addition, the complexity of the DEJ layer makes it a major challenge in the field of anti-aging research.

[0004] A Chinese patent (CN118873431A) discloses a polypeptide composition that promotes skin repair, including palmitoyl tripeptide-5, acetyl hexapeptide-37, snow lotus stem cell extract, etc., which can synergistically enhance the skin barrier function and is suitable for sensitive skin repair. However, this solution does not disclose the repair effect on skin DEJ. Summary of the Invention

[0005] The purpose of the present invention is to provide a polypeptide complex and its application, which has the advantages of small molecular weight, high safety, low allergy risk, strong permeability, and excellent repair and anti-aging functions.

[0006] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0007] The present invention provides a polypeptide complex, which contains a polypeptide Pep-IV with an amino acid sequence as shown in SEQ ID NO.1, a polypeptide Pep-VII with an amino acid sequence as shown in SEQ ID NO.2, and a polypeptide Pep-XVII with an amino acid sequence as shown in SEQ ID NO.3.

[0008] Preferably, the weight ratio of polypeptide Pep-IV, polypeptide Pep-VII and polypeptide Pep-XVII in the polypeptide complex is 1:0.8~1.2:0.8~1.2.

[0009] The present invention also provides the use of the polypeptide complex in preparing a product for repairing the basement membrane of the skin.

[0010] The present invention also provides the use of the polypeptide complex in the preparation of anti-aging products.

[0011] The present invention also provides the use of the polypeptide complex in the preparation of a collagen synthesis promoter.

[0012] Preferably, the collagen includes COL4A1, COL7A1, COL17A1, COL6A2, COL8A1, COL14A1 and COL16A1.

[0013] The present invention also provides the use of the polypeptide complex in preparing a product for inhibiting skin inflammation.

[0014] The present invention also provides the use of the polypeptide complex in preparing a product for promoting DNA damage repair or a skin cell cycle protein regulating agent.

[0015] The present invention also provides the use of the above polypeptide complex in preparing a LAMA2 gene expression promoting reagent or an MMP19 gene expression inhibiting reagent.

[0016] The present invention also provides a skin care product containing the polypeptide complex.

[0017] Beneficial effects of the present invention:

[0018] The present invention synergistically formulates three specific polypeptides to produce a polypeptide complex that has the efficacy of targeted repair of the skin's basement membrane. This polypeptide complex has the advantages of a small molecular weight, high safety, low allergy risk, and strong permeability, and has excellent repair and anti-aging functions. Experiments have shown that the polypeptide complex provided by the present invention, as an active peptide raw material, can significantly promote the production of endogenous IV, VII, and XVII collagen in the skin, and is also beneficial for repairing the skin's extracellular matrix, resisting DNA damage, improving skin cyclins, inhibiting the release of inflammatory factors, etc., and can effectively repair the basement membrane, with broad application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1A The effects of different ratios of Pep-IV, Pep-VII, and Pep-XVII on DEJ COL4A1 the impact of;

[0020] Figure 1BThe effects of different ratios of Pep-IV, Pep-VII, and Pep-XVII on DEJ COL7A1 the impact of;

[0021] Figure 1C The effects of different ratios of Pep-IV, Pep-VII, and Pep-XVII on DEJ COL17A1 the impact of;

[0022] Figure 2A The peptide complex and the same sequence of single collagen in the epidermis-dermis junction COL4A1 the impact of;

[0023] Figure 2B The peptide complex and the same sequence of single collagen in the epidermis-dermis junction COL7A1 the impact of;

[0024] Figure 2C The peptide complex and the same sequence of single collagen in the epidermis-dermis junction COL17A1 the impact of;

[0025] Figure 3 Peptide complex LAMA2 Effects on gene expression;

[0026] Figure 4 Peptide complex IL-11 Effects on gene expression;

[0027] Figure 5 These are the human body Raman permeability test results of the peptide complex. DETAILED DESCRIPTION

[0028] The present invention provides a polypeptide complex comprising a polypeptide Pep-IV having an amino acid sequence as shown in SEQ ID NO. 1, a polypeptide Pep-VII having an amino acid sequence as shown in SEQ ID NO. 2, and a polypeptide Pep-XVII having an amino acid sequence as shown in SEQ ID NO. 3. Preferably, the weight ratio of the polypeptides Pep-IV, Pep-VII, and Pep-XVII in the polypeptide complex is 1:0.8-1.2:0.8-1.2.

[0029] The present invention also provides the use of the aforementioned polypeptide complex in the preparation of a product for repairing the skin's basement membrane. In this invention, the "skin basement membrane repair product" refers to a skin care or medical product that repairs or strengthens the basement membrane structure (skin basement membrane) at the junction of the epidermis and dermis. By restoring the basement membrane's function, it improves skin problems caused by basement membrane damage. In this invention, the dosage form of the product can be selected from serums, lotions, creams, gels, sprays, ointments, patches, lyophilized powders, oils, microneedles, rollerballs, ampoules, mud masks, creams, aerosols, or nano-microemulsions.

[0030] The present invention also provides the use of the above polypeptide complex in the preparation of anti-aging products. In the present invention, the "anti-aging product" refers to a skin care preparation used to delay the signs of skin aging, and the product may also include active compound ingredients, excipients, etc.

[0031] The present invention also provides the use of the above-mentioned polypeptide complex in the preparation of a collagen synthesis promoter. In the present invention, the "collagen synthesis promoter" refers to a preparation that can increase the amount of collagen produced in the skin, and its dosage form can be selected from gels, freeze-dried powders, microneedles, or roller balls.

[0032] The present invention also provides the use of the above-mentioned polypeptide complex in the preparation of a product for inhibiting skin inflammation. In the present invention, the "product for inhibiting skin inflammation" refers to a care preparation for alleviating skin inflammatory reactions, and its dosage form can be selected from creams, patches, sprays, or facial masks.

[0033] The present invention also provides the use of the aforementioned polypeptide complex in the preparation of a product for promoting DNA damage repair or a skin cell cycle protein regulatory agent. In the present invention, the "product for promoting DNA damage repair" refers to a preparation used to repair DNA damage in skin cells or in vitro cultured cells, and the "skin cell cycle protein regulatory agent" refers to a preparation that regulates the expression of cell cycle-related proteins. The dosage form can be selected from lyophilized powder, aerosol, nano-microemulsion, or oil.

[0034] The present invention also provides the above polypeptide complex in the preparation LAMA2 Gene expression promoting agents or MMP19 In the present invention, the " LAMA2 Gene expression promoting reagents are used to increase LAMA2 Preparation of gene expression levels," MMP19 Gene expression inhibitory agents are used to reduce MMP19 The preparation of gene expression level can be used for gene expression level regulation in related experiments.

[0035] The present invention also provides a skin care product comprising the polypeptide complex. In the present invention, the skin care product can be in the form of an essence, lotion, cream, gel, or freeze-dried powder, and the compounded ingredients can be selected from moisturizers, antioxidants, emollients, soothing agents, film formers, preservatives, emulsifiers, pH adjusters, penetration enhancers, mineral components, amino acids, sugar derivatives, or lipid components.

[0036] The technical solutions provided by the present invention are described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0037] Example 1

[0038] The polypeptide Pep-IV with an amino acid sequence as shown in SEQ ID NO.1, the polypeptide Pep-VII with an amino acid sequence as shown in SEQ ID NO.2, and the polypeptide Pep-XVII with an amino acid sequence as shown in SEQ ID NO.3 were mixed in a weight ratio of 1:1:1 to obtain a polypeptide complex.

[0039] Example 2

[0040] The polypeptide Pep-IV with an amino acid sequence as shown in SEQ ID NO.1, the polypeptide Pep-VII with an amino acid sequence as shown in SEQ ID NO.2, and the polypeptide Pep-XVII with an amino acid sequence as shown in SEQ ID NO.3 were mixed in a weight ratio of 1:0.8:1.2 to obtain a polypeptide complex.

[0041] Example 3

[0042] The polypeptide Pep-IV with an amino acid sequence as shown in SEQ ID NO.1, the polypeptide Pep-VII with an amino acid sequence as shown in SEQ ID NO.2, and the polypeptide Pep-XVII with an amino acid sequence as shown in SEQ ID NO.3 were mixed in a weight ratio of 1:1.2:0.8 to obtain a polypeptide complex.

[0043] Experimental example

[0044] Pep-IV, Pep-VII, and Pep-XVII were obtained through database selection, and their sequences are shown in Table 1 below:

[0045] Table 1 Sequences of Pep-IV, Pep-VII, and Pep-XVII

[0046]

[0047] The researchers investigated the effects of this peptide complex on collagen types IV, VII, and XVII at a mass ratio of (Pep-IV: Pep-VII: Pep-XVII) = (10%-40%): (10%-70%): (15%-50%). Ultimately, they identified a unique ratio (Pep-IV: Pep-VII: Pep-XVII) = 1:1:1 that simultaneously promoted the growth of all three key collagens in DEJ. This resulting peptide complex was named the peptide complex.

[0048] The specific testing process is as follows:

[0049] RT-qPCR (real-time quantitative polymerase chain reaction) COL4A1 、 COL7A1 and COL17A1 Gene expression:

[0050] HaCaT cells in the logarithmic growth phase were cultured at a rate of 1×10 6 Cells were seeded into 6-well plates and cultured in a 37°C 5% CO2 incubator for 24 h. The culture medium was discarded, and the sample group was added with samples (mixed in a set ratio, with a final total peptide concentration of 10 ppm, 10 mg / L, the same below). The control group and the model group were added with serum-free culture medium and cultured in a 37°C 5% CO2 incubator for 24 h. The culture medium was discarded, and UVB (ultraviolet B band, 60 mJ / cm 2 ) were treated with UVB (the blank control group did not require UVB treatment). After treatment, serum-free culture medium was added and the cells were cultured at 37°C in a 5% CO2 incubator for 24 h. The supernatant was discarded, and the cells were washed twice with PBS and collected. RNA was extracted, reverse transcribed into cDNA, and then RT-qPCR was performed.

[0051] The experimental results are as follows Figure 1A to Figure 1C As shown:

[0052] After UVB irradiation, HaCaT cells in the model group (NC) COL4A1, COL7A1, COL17A1 The relative expression of genes was significantly reduced, indicating that the model was effective. When (Pep-IV: Pep-VII: Pep-XVII) = 1:1:1 (the final concentration of total peptide was 10ppm), the composite peptide had an COL4A1, COL7A1, COL17A1 The relative gene expression levels increased by 34.8%, 49.38%, and 75.77%, respectively. This is the only ratio in the test that can significantly promote the expression of three collagen genes in DEJ at the same time. The complex polypeptide under this ratio is named polypeptide complex.

[0053] The results of a comparative study on the promoting effects of the peptide complex and single collagen with the same sequence on the three types of collagen in DEJ showed that the peptide complex had a better promoting effect on type IV, type VII, and type XVII collagen than Pep-IV, Pep-VII, and Pep-XVII, and had a synergistic effect.

[0054] The specific testing process is as follows:

[0055] RT-qPCR assay COL4A1 、 COL7A1 and COL17A1 Gene expression:

[0056] HSF cells in logarithmic growth phase were cultured at a rate of 1×10 6 Cells were seeded into 6-well plates and cultured in a 37°C 5% CO2 incubator for 24 h. The culture medium was discarded, and samples were added to the sample group (the final concentration of the single sample and the mixed sample was 10 ppm, respectively; the final concentration of the Yangshen sample was 100 ppm VC + 7 ppm VE). Serum-free culture medium was added to the control group and the model group, and cultured in a 37°C 5% CO2 incubator for 24 h. The culture medium was discarded, and UVB (60 mJ / cm 2 ) were treated with UVB (the blank control group did not require UVB treatment). After treatment, serum-free culture medium was added and the cells were cultured at 37°C in a 5% CO2 incubator for 24 h. The supernatant was discarded, and the cells were washed twice with PBS and collected. RNA was extracted, reverse transcribed into cDNA, and then RT-qPCR was performed.

[0057] The experimental results are as follows Figure 2A to Figure 2C As shown (1:1:1-DEJ in the figure is the polypeptide complex of the present invention, and subsequent experimental results Figure 3 and Figure 4 1:1:1-DEJ in (same as above):

[0058] After UVB irradiation, HSF cells in the model group (NC) COL4A1 、 COL7A1 、 COL17A1 The relative expression of genes was significantly reduced, indicating that the model was effective. COL4A1 、 COL7A1 、 COL17A1 The relative gene expression levels increased by 27.01%, 49.48%, and 84.70%, respectively. The promotion effects were significantly better than those of Pep-IV, Pep-VII, and Pep-XVII, indicating that the polypeptide complex is beneficial for targeting and strengthening DEJ and promoting the expression of three collagen proteins in DEJ.

[0059] Peptide complex LAMA2 Effects of gene expression

[0060] Using a co-culture system of keratinocytes after sample incubation and fibroblasts stimulated by UVA (ultraviolet A band) + UVB, based on transcriptome sequencing analysis, the effects of the peptide complex on ECM (extracellular matrix) protein-related genes in skin cell signal transduction were studied. LAMA2 is a key gene in the laminin family and one of the main components of the extracellular matrix (ECM). It plays an important role in maintaining basement membrane structure, cell attachment and signal transduction, promoting cell migration and tissue repair, regulating immune response and inflammation, etc. The results showed that the peptide complex can significantly promote LAMA2 The gene expression was promoted by 102.26%, and the effect was better than that of a single collagen with the same sequence.

[0061] The specific testing process is as follows:

[0062] (1) Cell seeding: After the cells were revived, when the plating rate reached about 60%, keratinocytes (HaCaT) were seeded into Transwell cell culture plates, and fibroblasts were seeded into 6-well plates. The cells were incubated overnight in a CO2 incubator (37°C, 5% CO2). After the incubation, the fibroblasts were incubated for 7.5 h and then irradiated with UVA and UVB. The irradiation dose was UVA (30 J / cm 2 ) and UVB (50 mJ / cm 2 ), irradiation time 0.5 h. Keratinocytes (HaCaT) were subjected to subsequent drug administration procedures;

[0063] (2) Solution preparation: Prepare the working solution of the test substance according to the test group;

[0064] (3) Drug administration: According to the test group, when the cell plating rate in the Transwell cell culture plate reaches 40% to 60%, the group drug administration is carried out, and each group has 3 replicates. 1 mL of culture medium is added to each well of the blank control group and the negative control group, 1 mL of culture medium containing EGF (1 ppb, 1 μg / L) is added to each well of the positive control group 1, and 1 mL of culture medium containing retinol (10 μM) is added to each well of the positive control group 2. 1 mL of culture medium containing the corresponding concentration of the test sample (10 ppm) is added to each well of the sample group. After the drug administration is completed, the Transwell cell culture plate is placed in a CO2 incubator (37°C, 5% CO2) and cultured for 8 hours;

[0065] (4) Co-culture: Place the Transwell cell culture plate on a 6-well plate and culture in a CO2 incubator (37°C, 5% CO2) for 24 h;

[0066] (5) Transcriptome sampling: After incubation, discard the culture medium and collect the cells;

[0067] (6) After washing twice with PBS, RNA extraction and library construction were performed;

[0068] (7) Total RNA was extracted using TRIzol reagent according to the manufacturer's instructions. RNA purity and quantification were determined using a NanoDrop 2000 spectrophotometer (Thermo Scientific, USA), and RNA integrity was assessed using an Agilent 2100 Bioanalyzer (Agilent Technologies, USA). Transcriptome libraries were constructed using the VAHTS Universal V5 RNA-seq Library Prep Kit according to the manufacturer's instructions.

[0069] (8) The library was sequenced and data processed using the Illumina Novaseq 6000 sequencing platform. HISAT2 software was used for reference genome alignment and gene expression (FPKM) calculation.

[0070] (9) Data screening criteria: Pairwise comparisons were performed between groups, and genes with a p value < 0.05 were selected as candidate differentially expressed genes, and their gene functions were further investigated.

[0071] The experimental results are as follows Figure 3 As shown:

[0072] Compared with the blank group (BC), the ECM protein-related genes in the model group (NC) LAMA2 Compared with the model group (NC), the peptide complex significantly promoted the expression of LAMA2 gene by 102.26%, and the effect was better than that of a single collagen with the same sequence.

[0073] Effects of polypeptide complexes on the expression of multiple collagen genes

[0074] The effects of the peptide complex on the expression of multiple collagen genes were studied based on transcriptome sequencing analysis using a co-culture system of keratinocytes after sample incubation and fibroblasts after combined UVA+UVB stimulation. COL6A2, COL7A1, COL8A1, COL14A1, COL16A1 The protein encoded by the gene is collagen, one of the main components of the skin, which gives the skin its strength, elasticity and firmness. MMP19 The protein encoded by this gene is an enzyme belonging to the matrix metalloproteinase (MMP) family. The catalytic domain of MMP-19 is capable of hydrolyzing basement membrane components including type IV collagen, laminin, and collagen, as well as the large tenascin C isoform, fibronectin, and type I gelatin. This indicates that MMP-19 is a potent basement membrane degrader capable of hydrolyzing multiple extracellular matrix components. The specific testing procedure is the same as above.

[0075] The experimental results are shown in Table 2:

[0076] Compared with the BC group, the NC group COL6A2, COL8A1, COL14A1, COL16A1 The relative gene expression levels decreased by 16.28%, 162.84%, 171.19%, and 42.64%, respectively. MMP19 The relative gene expression increased by 34.78%, indicating that the model is effective. Compared with the NC group, the polypeptide complex significantly promoted COL6A2, COL8A1, COL14A1, COL16A1 The relative gene expression levels increased by 18.86%, 64.71%, 96.29%, and 27.33%, respectively. MMP19 The relative expression of the gene decreased by 29.08%. The peptide complex promoted collagen expression and inhibited collagen degradation better than single collagen of the same sequence.

[0077] Table 2 Effects of the peptide complex on various collagen-related genes

[0078]

[0079] *The expression ratio is the ratio of expression levels between two treatment groups. An expression ratio > 1 indicates an increase in expression level, and an expression ratio < 1 indicates a decrease in expression level. COL4, COL7, and COL17 refer to Pep-IV, Pep-VII, and Pep-XVII screened in Example 1, respectively.

[0080] Effects of polypeptide complexes on DNA damage repair genes

[0081] The effects of the peptide complex on DNA damage repair genes were studied based on transcriptome sequencing analysis using a co-culture system of keratinocytes after sample incubation and fibroblasts after UVA+UVB combined stimulation. NEIL3 The gene encodes DNA glycosylase, which initiates the first step of base excision repair by cleaving bases damaged by reactive oxygen species and introducing DNA chain breaks through related cleavage enzyme reactions. LIG1 The gene encodes a member of the ATP-dependent DNA ligase protein family, which plays a role in DNA replication, recombination, and base excision repair. NEIL3 Genes and LIG1 All genes belong to the base excision repair signaling pathway. MLH1 Gene, MSH6 Gene, MSH2 Gene, RFC1 Gene, RFC2 Gene, RFC3 Gene, RFC4 Gene, RFC5 Gene, EXO1 Gene, RPA2 All genes belong to the mismatch repair signaling pathway. The specific testing process is the same as above.

[0082] The experimental results are shown in Table 3:

[0083] Compared with the BC group, the NC group NEIL3 and LIG1 The relative expression of genes in the NC group and the control group decreased by 133.10% and 98.42%, respectively, indicating that the model is effective. NEIL3 and LIG1 The relative expression levels of genes in the NC group increased by 62.90% and 72.38% respectively. MLH1 Gene, MSH6 Gene, MSH2 Gene, RFC1 Gene, RFC2 Gene, RFC3 Gene, RFC4 Gene, RFC5 Gene, EXO1 Genes and RPA2 The relative gene expression levels decreased by 23.98%, 71.86%, 96.97%, 34.07%, 86.70%, 123.66% and 104.43%, 82.89%, 95.78% and 70.89% respectively, indicating that the model is effective. MLH1 Gene, MSH6 Gene, MSH2 Gene, RFC1 Gene, RFC2 Gene, RFC3 Gene, RFC4 Gene, RFC5 Gene, EXO1 Genes and RPA2 The relative expression levels of the genes increased by 14.24%, 52.29%, 52.69%, 22.20%, 56.29%, 66.44%, 51.82%, 47.68%, 67.08% and 40.58%, respectively. The polypeptide complex has a better effect on promoting DNA damage repair genes than single collagen with the same sequence.

[0084] Table 3 Effects of polypeptide complexes on DNA damage repair genes

[0085]

[0086] *The expression ratio is the ratio of expression levels between two treatment groups. An expression ratio > 1 indicates an increase in expression level, and an expression ratio < 1 indicates a decrease in expression level. COL4, COL7, and COL17 refer to Pep-IV, Pep-VII, and Pep-XVII screened in Example 1, respectively.

[0087] Effects of peptide complexes on cyclin-related genes

[0088] The effects of the peptide complex on cyclin-related genes were studied based on transcriptome sequencing analysis using a co-culture system of keratinocytes after sample incubation and fibroblasts after UVA+UVB combined stimulation. CDK4 and CDK2 Gene encoding a cell cycle-dependent kinase, belonging to CDK The CCNE2 family is a key factor in cell cycle regulation, particularly in the G1 / S phase transition. The protein encoded by the CCNE2 gene belongs to a highly conserved family of cyclins, whose members are characterized by a significant periodicity in protein abundance throughout the cell cycle. CCNA2 The gene encodes A-type cyclin A2, a member of the cyclin family. The specific testing process is the same as above.

[0089] The experimental results are shown in Table 4:

[0090] Compared with the BC group, the NC group CDK4, CCNE2, CDK2, CCNA2 The relative expression levels of genes were reduced by 30.56%, 158.97%, 124.72% and 142.98% respectively, indicating that the model is effective. CDK4, CCNE2, CDK2, CCNA2 The relative gene expression levels increased by 26.30%, 115.53%, 71.81% and 54.38%, respectively.

[0091] Table 4 Effects of the peptide complex on cyclin-related genes

[0092]

[0093] *The expression ratio is the ratio of expression levels between two treatment groups. An expression ratio > 1 indicates an increase in expression level, and an expression ratio < 1 indicates a decrease in expression level. COL4, COL7, and COL17 refer to Pep-IV, Pep-VII, and Pep-XVII screened in Example 1, respectively.

[0094] Effects of peptide complex on IL-11 gene

[0095] The effects of peptide complexes on the expression of keratinocytes after sample incubation and fibroblasts after UVA+UVB combined stimulation were studied based on transcriptome sequencing analysis. IL-11 Genetic influence. IL-11 is a proinflammatory cytokine in the IL-6 family. Inhibiting the signaling of the proinflammatory protein IL-11 can significantly delay aging in mammals. In certain inflammatory environments, IL-11 can stimulate fibroblasts to secrete inflammatory mediators such as IL-8 and VEGF, promoting angiogenesis and the recruitment of inflammatory cells, thereby exacerbating the inflammatory response. The specific testing process is the same as above.

[0096] The experimental results are as follows Figure 4 As shown:

[0097] Compared with the BC group, the NC group IL-11 The relative expression of genes increased significantly by 74.83%, indicating that the model is effective. IL-11 The relative expression of the gene decreased significantly by 30.05%, indicating that the polypeptide complex is beneficial in inhibiting skin inflammation and its effect is better than that of a single peptide with the same sequence.

[0098] Human Raman permeability test results of peptide complexes

[0099] The front end of the human forearm was selected for human Raman testing: the sample used for the test was DEJ Booter, and the application amount was 5 mg / cm 2 , select 1×1 cm 2 The test skin area was 1 × 1 cm. The LabRAM Odyssey high-speed, high-resolution confocal Raman microscope (HORIBA) was used to perform the test at 0 h, 0.5 h, 2 h, 4 h, and 6 h. The test was conducted on one person and the test area was 3 1 × 1 cm 2 The skin area tested was sized and 5 points were selected within each test area for 3 parallel tests. The relative permeability of the peptide complex was calculated.

[0100] Raman spectroscopy imaging data processing includes spectral preprocessing and data analysis. Spectral preprocessing includes cosmic ray removal, spectral smoothing, background noise removal, baseline calibration, and spectral normalization. Univariate data analysis primarily analyzes the Raman spectral data for biochemical substances corresponding to specific peak positions, revealing the distribution of these substances within human skin.

[0101] Data analysis: Labspec software was used to perform baseline calibration of the Raman spectra and confirm the positions of characteristic peaks. The obtained Raman spectra were calculated, including peak intensity, peak shift, peak area, half-peak width, etc. Labspec software was also used to perform numerical analysis of the peak intensities corresponding to different depths and to plot their spatial distribution.

[0102] The obtained Raman spectrum is first pre-processed and the initial position of the Raman signal is used as the 0 μm position on the skin surface. After feature selection, the keratin / lipid related characteristic information is selected to determine the junction of the stratum corneum and the active epidermis and the junction of the active epidermis and the dermis with the change of the depth in the skin and the change of the water content in the skin, so as to confirm the information related to the skin depth. The characteristic Raman signal of the product is then used to confirm its distribution in different skin depth spaces, which is different from the intrinsic signal of the skin, so as to describe the permeability behavior of the product. The data normal distribution test (Shapiro-Wilk Test) is performed using software. If it is normally distributed, paired t Otherwise, the two-sample rank sum test is used.

[0103] Relative penetration rate (%) = Raman spectrum intensity of active substance entering the skin / Raman spectrum intensity of active substance applied to the skin * 100%

[0104] The experimental results are as follows Figure 5 As shown:

[0105] After using the peptide complex on human skin, the relative permeability of the peptide complex was 13.57%, 30.88%, 52.35% and 60.45% within 0.5 h, 2 h, 4 h and 6 h, respectively. It can reach the DEJ layer within 2 h of use, effectively strengthening the skin basement membrane.

[0106] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as within the scope of protection of the present invention.

Claims

1. A polypeptide complex, characterized in that The polypeptide complex contains a polypeptide Pep-IV with an amino acid sequence as shown in SEQ ID NO.1, a polypeptide Pep-VII with an amino acid sequence as shown in SEQ ID NO.2, and a polypeptide Pep-XVII with an amino acid sequence as shown in SEQ ID NO.3; The weight ratio of polypeptide Pep-IV, polypeptide Pep-VII and polypeptide Pep-XVII in the polypeptide complex is 1:1:

1.

2. Use of the polypeptide complex according to claim 1 in preparing a product for repairing the basement membrane of the skin.

3. Use of the polypeptide complex according to claim 1 in the preparation of anti-aging products.

4. Use of the polypeptide complex according to claim 1 in the preparation of a collagen synthesis promoter.

5. The use according to claim 4, characterized in that The collagen synthesis promoter promotes collagen synthesis by increasing the expression of collagen synthesis genes, including COL4A1, COL7A1, COL17A1, COL6A2, COL8A1, COL14A1 and COL16A1.

6. Use of the polypeptide complex according to claim 1 in preparing a product for inhibiting skin inflammation.

7. A skin care product, characterized in that: Contains the polypeptide complex according to claim 1.

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