Collagen xvii cleavage peptide and its use in hair loss products

Abstract

The application provides a type XVII collagen truncated peptide and application thereof in hair loss prevention products, and belongs to the technical field of peptide products. The type XVII collagen truncated peptide provided by the application can significantly improve the antioxidant capacity of hair follicles, effectively enhance the structural stability of hair follicles, strongly strengthen the hair root toughness, and obviously delay the aging process of scalp cells, thereby synergistically improving the hair loss problem from multiple dimensions. The product provided by the application replaces large proteins with small molecule peptides, solves the technical obstacles of traditional collagen, such as difficult transdermal absorption and easy degradation, and provides a new solution for active ingredients for preventing hair loss. The application provides a core ingredient with high activity, easy absorption and strong safety for the field of preventing hair loss, and meets the urgent needs of consumers for high-efficiency hair loss prevention products. The application realizes the integration of the triple functions of "preventing hair loss, fixing hair and resisting aging" by using industrialized peptide technology, and promotes the upgrading of hair loss prevention products to high efficiency, convenience and universality.

Description

Technical Field

[0001] The present invention relates to the technical field of peptide products, in particular to type XVII collagen truncated peptides and applications thereof in anti-hair loss products. Background Art

[0002] Hair follicle stem cells are primarily located around the bulge and dermal papilla of the hair follicle, maintaining cyclical hair regeneration. While typically dormant within the follicle, these cells rapidly activate and divide during the hair growth cycle, generating hair or repairing damaged areas of the follicle. Therefore, the vitality of these stem cells is a crucial indicator of the follicle's ability to regenerate hair. Currently, a core mechanism of hair loss is dysfunction of these stem cells, such as when their activity is suppressed, leading to prolonged dormancy, or when stem cells differentiate directly into epidermal keratinocytes rather than hair follicle cells.

[0003] Studies have shown that with aging, COL17A1 expression decreases, leading to a disruption of the hair follicle stem cell niche and stem cell loss, a key mechanism underlying hair thinning and graying. COL17A1 collagen is a key component of hemidesmosomes, helping anchor stem cells in the basal layer of the epidermis to the basement membrane and maintaining the stability of the microenvironment of hair follicle stem cells (located in the bulge). COL17A1 maintains stem cell self-renewal and inhibits differentiation by stabilizing the Wnt signaling pathway (Fzd4, Nfatc2, Nfatc4, and Tcf7, among others). Therefore, supplementing with type XVII collagen is a viable method for maintaining the differentiation activity of hair follicle stem cells and a viable anti-hair loss strategy.

[0004] However, type XVII collagen is a large protein at 295.2 kDa, limiting its application in absorbable topical cosmetics. Truncated functional peptides are an important form of collagen in cosmetic applications. These peptides have a smaller molecular weight, resulting in improved transdermal absorption and absorption, as well as high stability and formulation compatibility. Therefore, functional type XVII collagen peptides hold great potential for use in daily anti-hair loss products. Summary of the Invention

[0005] The purpose of the present invention is to provide a type XVII collagen truncated peptide and its application in anti-hair loss products, thereby providing an effective means for the development of daily anti-hair loss products.

[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 an application of a type XVII collagen truncated peptide in the preparation of an anti-hair loss product. The amino acid sequence of the type XVII collagen truncated peptide is shown in SEQ ID NO.1.

[0008] The present invention also provides a use of a type XVII collagen truncated peptide in the preparation of a hair follicle stem cell activator. The amino acid sequence of the type XVII collagen truncated peptide is shown in SEQ ID NO.1.

[0009] The present invention also provides a use of a type XVII collagen truncated peptide in the preparation of an antioxidant scalp care product, wherein the amino acid sequence of the type XVII collagen truncated peptide is shown in SEQ ID NO.1.

[0010] The present invention also provides a use of a type XVII collagen truncated peptide in preparing a product for inhibiting scalp cell aging. The amino acid sequence of the type XVII collagen truncated peptide is shown in SEQ ID NO.1.

[0011] The present invention also provides a use of a type XVII collagen truncated peptide in preparing a gene expression regulator, wherein the gene is selected from at least one of COL17A1, ITGβ1, ITGA6, NES, and SOX10, and the amino acid sequence of the type XVII collagen truncated peptide is shown in SEQ ID NO.1.

[0012] The present invention also provides an application of a type XVII collagen truncated peptide in the preparation of a product for promoting TGF-β, ALPL, NOG or CD44 gene expression, wherein the amino acid sequence of the type XVII collagen truncated peptide is shown in SEQ ID NO.1.

[0013] The present invention also provides an external product for preventing hair loss, which comprises a truncated peptide of type XVII collagen as an active ingredient, wherein the amino acid sequence of the truncated peptide of type XVII collagen is shown in SEQ ID NO.1;

[0014] The dosage form of the anti-hair loss external product includes shampoo, essence, emulsion or spray.

[0015] The present invention also provides an anti-hair loss composition comprising a truncated peptide of type XVII collagen, wherein the amino acid sequence of the truncated peptide of type XVII collagen is shown in SEQ ID NO.1.

[0016] Preferably, the anti-hair loss composition further comprises minoxidil and / or finasteride.

[0017] Beneficial effects of the present invention:

[0018] The type XVII collagen truncated peptide provided by the present invention can significantly enhance the antioxidant capacity of hair follicles, effectively strengthen the structural stability of hair follicles, strongly strengthen the toughness of hair roots, and significantly delay the aging process of scalp cells, thereby synergistically improving the problem of hair loss from multiple dimensions. The product provided by the present invention replaces large proteins with small molecule peptides, solving the technical obstacles of traditional collagen, which is difficult to be absorbed through the skin and easy to degrade, and provides a new solution for anti-hair loss active ingredients; the active ingredients provided by the present invention can be flexibly developed into daily care products such as shampoo, essence, spray, or integrated into medical-grade devices such as microneedle patches and hair growth devices, providing a highly active, easily absorbed, and safe core ingredient for the anti-hair loss field, meeting consumers' urgent demand for efficient anti-hair loss products; using industrializable peptide technology, the triple efficacy of "anti-hair loss-strengthening-anti-aging" is achieved, promoting the upgrading of anti-hair loss products to be more efficient, convenient, and universal. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 This is the result diagram of the effect of Hair-Col17 on the expression of antioxidant-related genes;

[0020] Figure 2 This is a graph showing the effect of Hair-Col17 on the expression of genes related to hair follicle stabilization;

[0021] Figure 3 This is the result diagram of the effect of Hair-Col17 on the expression of genes related to strong hair roots;

[0022] Figure 4 This figure shows the effect of Hair-Col17 on senescent hair follicle cells. DETAILED DESCRIPTION

[0023] The present invention is based on a truncated peptide of type XVII collagen, the amino acid sequence of which is GRGDNP, as shown in SEQ ID NO. 1. In the present invention, the truncated peptide refers to a functional short peptide obtained by selectively designing and cleaving type XVII collagen, and has the excellent properties of enhanced transdermal absorption, improved stability, and retained hair follicle bioactivity.

[0024] The present invention provides an anti-hair loss composition comprising the type XVII collagen truncated peptide provided herein. Preferably, the anti-hair loss composition further comprises minoxidil and / or finasteride. The anti-hair loss composition may further comprise plant-based anti-hair loss active ingredients (such as Platycladus orientalis extract and ginsenosides), vitamin-based hair growth factors (such as biotin and niacinamide), trace elements (such as zinc compounds), penetration enhancers (such as azone and lecithin), and scalp conditioning agents (such as panthenol and ceramide).

[0025] The present invention also provides the use of the aforementioned type XVII collagen truncated peptide in the preparation of anti-hair loss products. In the present invention, such anti-hair loss products may include daily care products (e.g., anti-hair loss shampoos, hair growth essences), medical devices (e.g., hair growth device accessories, microneedle patches), professional line products (e.g., scalp perfusion solutions, anti-hair loss treatment kits), and household instrument consumables (e.g., hair growth comb infusion solutions).

[0026] The present invention also provides the use of the aforementioned type XVII collagen truncated peptide in the preparation of a hair follicle stem cell activator. The activator may further comprise stem cell proliferation cofactors (such as EGF and FGF), hair follicle microenvironment regulators (such as Wnt pathway activators), cell energy metabolism promoters (such as ATP precursors), and stem cell protective components (such as mitochondrial antioxidants).

[0027] The present invention also provides the use of the aforementioned truncated type XVII collagen peptide in the preparation of an antioxidant scalp care product. The product may also incorporate free radical scavengers (such as vitamin C derivatives and glutathione), photodamage repair ingredients (such as ectoine and melatonin), oxidative stress relievers (such as ergothioneine and astaxanthin), and barrier-repairing lipids (such as squalane and cholesterol).

[0028] The present invention also provides the use of the aforementioned truncated type XVII collagen peptide in the preparation of a product for inhibiting scalp cell aging. These products can be supplemented with cellular aging inhibitors (such as senolytics and NAD+ precursors), telomere-protecting factors (such as telomerase activators), DNA repair agents (such as photolyases), and inflammation-reducing ingredients (such as bisabolol and dipotassium glycyrrhizate).

[0029] The present invention also provides the use of the aforementioned type XVII collagen truncated peptide in the preparation of a gene expression regulator, wherein the gene is selected from at least one of COL17A1, ITGβ1, ITGA6, NES, and SOX10. The regulator may also include gene transcription enhancing cofactors (such as histone deacetylase inhibitors), signaling pathway co-activators (such as Notch pathway regulators), epigenetic modification components (such as methyl donors), and gene delivery vectors (such as lipid nanoparticles).

[0030] The present invention also provides the use of the aforementioned type XVII collagen truncated peptide in the preparation of a product for promoting TGF-β, ALPL, NOG, or CD44 gene expression. Such a product can be combined with a hair follicle growth cycle regulator (such as a prostaglandin analogue), an extracellular matrix synthesis promoter (such as a silicon compound), a dermal papilla cell activator (such as adenosine), and an ingredient that improves hair follicle blood supply (such as a minoxidil derivative).

[0031] The present invention also provides a topical anti-hair loss product containing the type XVII collagen truncated peptide provided herein as an active ingredient. The dosage forms of the topical anti-hair loss product include shampoo, serum, lotion, or spray. The dosage forms can also be expanded to include gels, foams, roll-ons, ampoule concentrates, ointments, scalp scrubs, conditioners, hair masks, hair growth pens, and dissolvable microneedle arrays. Product excipients may include thickeners (such as carbomer), emulsifiers (such as PEG-40 hydrogenated castor oil), preservatives (such as phenoxyethanol), pH adjusters (such as lactic acid), and sensory enhancers (such as menthol).

[0032] 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.

[0033] Example

[0034] The sequence of the type XVII collagen shedding-preventing peptide provided by the present invention is GRGDNP, as shown in SEQ ID NO.1.

[0035] The short peptide was synthesized using conventional methods and used as a test sample to conduct the following experiments:

[0036] Type XVII collagen peptide (Hair-Col17) antioxidant-related gene expression test

[0037] The dermal papilla cells in the logarithmic growth phase were divided into 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 UVB (50 mJ / cm 2 ) (the blank control group did not require UVB treatment). Experimental groups were treated with samples at various concentrations, while the blank control and model groups were treated with serum-free culture medium and cultured in a 37°C, 5% CO2 incubator for 24 hours. The supernatant was discarded, and cells were washed twice with PBS before being harvested. RNA was extracted, reverse-transcribed into cDNA, and then subjected to RT-qPCR. Data are expressed as mean ± standard error. Differences between the control and negative control groups were analyzed using a T-test. Differences between the sample groups and the negative control group were analyzed using a one-way ANOVA. P < 0.05 (* and #) was considered significant, P < 0.01 (** and ##) was considered extremely significant, and P < 0.001 (*** and ###) was considered strongly significant.

[0038] Blank control (BC) group: cultured in serum-free medium;

[0039] Model (NC) group: 50 mJ / cm 2 Cultured in serum-free medium after UVB irradiation;

[0040] Experimental group: 50 mJ / cm 2 After UVB irradiation, the corresponding concentration of Hair-Col17 solution was added for culture;

[0041] Positive control group (PC): 50 mJ / cm 2 After UVB irradiation, 50 μg / mL finasteride solution was added to the cells for culture

[0042] like Figure 1 As shown, compared with the model group, Hair-Col17 significantly promoted the expression of antioxidant genes such as SOD2 and GPX3. Hair-Col17 at 5 ppm, 10 ppm, and 50 ppm increased SOD2 gene expression by 60.36% (P<0.001), 80.07% (P<0.001), and 99.70% (P<0.001), respectively. Hair-Col17 at 10 and 50 ppm significantly increased GPX3 gene expression by 143.75% (P<0.001) and 208.41% (P<0.001), respectively.

[0043] The results showed that 5, 10, and 50 ppm of Hair-Col17 had the effect of scavenging free radicals.

[0044] Type XVII collagen peptide (Hair-Col17) stabilizes the expression of hair follicle-related genes

[0045] The experimental procedure and experimental groups were the same as above.

[0046] like Figure 2 As shown in the figure, compared with the blank control group, the relative expression levels of COL17A1, ITGβ1, ITGA6, NES and SOX10 genes in the model group after UVB irradiation were significantly downregulated, indicating that the model was successfully established and the model group was effective.

[0047] Compared with the model group, the addition of Hair-Col17 after UVB stimulation can promote the expression of COL17A1, ITGβ1, ITGA6, NES and SOX10 genes related to hair follicle stability. 5, 50 ppm Hair-Col17 increased the expression of COL17A1 gene by 19.23% (P<0.01), 42.04% (P<0.001), respectively, while 50 μg / mL finasteride inhibited the expression of COL17A1 gene, and the expression was down-regulated by 30.99% (P<0.001). 5 ppm, 50 ppm Hair-Col17 increased the expression of ITGβ1 gene by 42.89% (P<0.01), 63.52% (P<0.001), respectively, while finasteride had no significant effect on the expression of ITGβ1 gene. 50 ppm Hair-Col17 and 50 μg / mL finasteride can promote the expression of ITGA6 gene, and the expression is increased by 60.17% (P<0.01), 66.71% (P<0.01), respectively. 5 ppm, 50 ppm Hair-Col17 increased the expression of NES gene by 68.99% (P<0.05), 253.55% (P<0.001), respectively, while finasteride had no significant effect on the expression of NES gene. 50 ppm Hair-Col17 increased the expression of SOX10 gene by 65.69% (P<0.05), while finasteride had no significant effect on the expression of SOX10 gene.

[0048] Expression test of Hair-Col17 on genes related to strong hair growth

[0049] The experimental process and experimental groups are the same as above.

[0050] As shown in Figure 3 Compared with the blank control group, the relative expression of TGF-β, ALPL, NOG and CD44 genes in the model group after UVB irradiation was significantly down-regulated, indicating that the modeling was successful, and the model group was effective.

[0051] Compared with the model group, the addition of Hair-Col17 after UVB stimulation promoted the expression of genes such as TGF-β, ALPL, NOG, and CD44, which are essential for hair root strength. Hair-Col17 at 10 and 50 ppm and finasteride at 50 μg / mL increased TGF-β gene expression by 333.30% (P < 0.001), 394.44% (P < 0.001), and 149.58% (P < 0.05), respectively. Hair-Col17 at 10 and 50 ppm increased ALPL gene expression by 52.66% (P < 0.05) and 93.66% (P < 0.001), respectively. Finasteride had no significant effect on ALPL gene expression. Hair-Col17 at 10 and 50 ppm, and finasteride at 50 μg / mL, increased NOG gene expression by 312.92% (P<0.001), 378.79% (P<0.001), and 130.32% (P<0.001), respectively. Hair-Col17 at 10 and 50 ppm, and finasteride at 50 μg / mL, increased CD44 gene expression by 94.29% (P<0.001), 145.22% (P<0.001), and 44.66% (P<0.01), respectively.

[0052] Senescent cell staining test

[0053] The dermal papilla cells in the logarithmic growth phase were divided into 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 UVB (50 mJ / cm 2 ) treated cells (the blank control group did not need UVB treatment); the experimental group was treated with samples of various concentrations, and the blank control group and model group were treated with serum-free culture medium, and cultured in a 37°C, 5% CO2 incubator for 24 hours; the supernatant was discarded, and the cells were washed twice with PBS, and 1 mL of β-galactosidase staining fixative was added and fixed at room temperature for 15 minutes; the cell fixative was aspirated, and the cells were washed three times with PBS for 3 minutes each time; the PBS was aspirated, and 1 mL of staining working solution was added to each well; photos were taken under a microscope, and the percentage of β-galactosidase-positive cells in the images was analyzed using software;

[0054] The relative content of β-galactosidase positive cells and senescent cells is calculated as follows:

[0055]

[0056] like Figure 4As shown in the figure, compared with the blank control group, the proportion of senescent cells in the model group increased after UVB irradiation, indicating that the model is effective. Compared with the model group, the addition of 5, 10, and 50 ppm Hair-Col17 and 50 μg / mL finasteride after UVB stimulation reduced the proportion of senescent cells, with cell senescence inhibition rates of 58.39% (P < 0.05), 63.59% (P < 0.01), 74.35% (P < 0.01), and 61.71% (P < 0.01), respectively.

[0057] As shown in the above examples, the expression of SOD2 and GPX3 genes in dermal papilla cells was analyzed by RT-qPCR to evaluate the free radical scavenging effect of Hair-Col17. Compared with the blank control group, the relative expression of SOD2 and GPX3 genes in the model group was significantly downregulated, indicating that the model was successfully established and the model group was effective. Compared with the model group, 5, 10, and 50 ppm Hair-Col17 and 50 μg / mL finasteride significantly increased SOD2 gene expression by 60.36% (P<0.001), 80.07% (P<0.001), 99.70% (P<0.001), and 41.44% (P<0.01), respectively. 10 and 50 ppm Hair-Col17 and 50 μg / mL finasteride significantly increased GPX3 gene expression by 143.75% (P<0.001), 208.41% (P<0.001), and 71.12% (P<0.05), respectively. These experimental results indicate that 5, 10, and 50 ppm Hair-Col17 has a free radical scavenging effect.

[0058] Using dermal papilla cells, the expression of genes associated with hair follicle stabilization, COL17A1, ITGβ1, ITGA6, NES, and SOX10, was measured by RT-qPCR to evaluate the effectiveness of Hair-Col17 in stabilizing hair follicles. Compared with the blank control group, the relative expression levels of COL17A1, ITGβ1, ITGA6, NES, and SOX10 were significantly downregulated in the model group, indicating that the model was successfully established and the model group was effective. Compared with the model group, 5 and 50 ppm Hair-Col17 increased the expression of COL17A1 gene by 19.23% (P<0.01) and 42.04% (P<0.001), respectively, while 50 μg / mL finasteride downregulated the expression of COL17A1 gene by 30.99% (P<0.001). 5 and 50 ppm Hair-Col17 increased the expression of ITGβ1 gene by 42.89% (P<0.001) and 63.52% (P<0.001), respectively, while finasteride had no significant effect on the expression of ITGβ1 gene. 50 ppm Hair-Col17 and 50 μg / mL finasteride increased the expression of ITGA6 gene by 60.17% (P<0.01) and 66.71% (P<0.01), respectively. Hair-Col17 increased NES gene expression by 68.99% (P<0.05) and 253.55% (P<0.001), respectively, while finasteride had no significant effect on NES gene expression. Hair-Col17 at 50 ppm increased SOX10 gene expression by 65.69% (P<0.05), while finasteride had no significant effect on SOX10 gene expression. These experimental results indicate that Hair-Col17 has a stabilizing effect on hair follicles.

[0059] The effect of Hair-Col17 on strengthening hair roots was evaluated by RT-qPCR analysis of the expression of genes associated with strong hair roots, including TGF-β, ALPL, CD44, and NOG, using dermal papilla cells. Compared to the blank control group, the relative expression of TGF-β, ALPL, NOG, and CD44 genes in the model group was significantly downregulated, indicating that the model was successfully established and effective. Compared with the model group, 10 and 50 ppm Hair-Col17 and 50 μg / mL finasteride increased the expression of TGF-β gene by 333.30% (P<0.001), 394.44% (P<0.001), and 149.58% (P<0.05), respectively. 10 and 50 ppm Hair-Col17 increased the expression of ALPL gene by 52.66% (P<0.05) and 93.66% (P<0.001), respectively, while finasteride had no significant effect on ALPL gene expression. 10 and 50 ppm Hair-Col17 and 50 μg / mL finasteride increased the expression of NOG gene by 312.92% (P<0.001), 378.79% (P<0.001), and 130.32% (P<0.001), respectively. Hair-Col17 and 50 μg / mL finasteride increased CD44 gene expression by 94.29% (P < 0.001), 145.22% (P < 0.001), and 44.66% (P < 0.01), respectively. These experimental results demonstrate that Hair-Col17 strengthens hair roots.

[0060] The anti-aging effect of Hair-Col17 was evaluated by measuring the proportion of senescent cells in dermal papilla cells using β-galactosidase staining. Compared with the blank control group, the proportion of senescent cells in the model group increased after UVB irradiation, indicating the effectiveness of the model. Compared with the model group, the inhibition rates of cell senescence at 5, 10, and 50 ppm Hair-Col17 and 50 μg / mL finasteride were 58.39% (P < 0.05), 63.59% (P < 0.01), 74.35% (P < 0.01), and 61.71% (P < 0.01), respectively. These experimental results demonstrate that Hair-Col17 has anti-aging effects.

[0061] 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 the scope of protection of the present invention.

Claims

1. A use of a type XVII collagen truncated peptide in the preparation of an anti-hair loss product, characterized in that: The amino acid sequence of the type XVII collagen truncated peptide is shown in SEQ ID NO.

1.

2. A use of a type XVII collagen truncated peptide in the preparation of an antioxidant scalp care product, characterized in that: The amino acid sequence of the type XVII collagen truncated peptide is shown in SEQ ID NO.

1.

3. An anti-hair loss composition comprising a truncated peptide of type XVII collagen, characterized in that: The amino acid sequence of the type XVII collagen truncated peptide is shown in SEQ ID NO.1; The anti-hair loss composition further comprises minoxidil and / or finasteride.

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