Use of an active peptide in the manufacture of a product to improve the skin barrier and promote collagen expression and / or increase skin elasticity and suppleness and / or reduce wrinkles

Abstract

Use of an active peptide in the manufacture of a product to improve the skin barrier and promote collagen expression and / or increase skin elasticity and suppleness and / or reduce wrinkles, characterized in that the active peptide has the structural formula HA-OH, wherein A comprises at least two amino acid residues, and the amino acid residues comprise at least one from Hyp, Gly, Pro and Ala.

Description

Technical field

[0001] The present invention belongs to the field of general food or functional food or pet food or pharmaceutical technology and specifically relates to the use of an active peptide in the manufacture of a product to improve the skin barrier and promote collagen expression and / or increase skin elasticity and suppleness and / or reduce wrinkles. State of the art

[0002] The skin is the body's largest organ, and the most obvious manifestation of aging is skin aging. The extracellular matrix (ECM) is a complex structure produced by cells and secreted to the cell surface or into the intercellular space. It consists of biological macromolecules and plays a crucial role in maintaining cell function and tissue integrity. The ECM is composed of various proteins such as collagen, glycoproteins, and proteoglycans, provides mechanical support, stores growth factors, and regulates cellular homeostasis.

[0003] Collagen is not only responsible for maintaining the morphological structure of tissues, but also plays a crucial role in various physiological and pathological processes. Its absence or alteration can have serious consequences for the body. Over 1,300 human diseases are known to be caused by collagen mutations. Currently, there is a growing body of research highlighting the relationship between collagen and the immune system.

[0004] Many natural active ingredients exert their anti-aging effects by improving the growth and metabolism of human skin cells, promoting increased cell activity, and enhancing the expression of the skin matrix. Due to their significant anti-aging effects, peptide substances have become key components in anti-aging products. However, current technology typically involves extracting collagen peptides from animal sources such as fish skin, pig skin, bovine hide, and tendons, and then using them to manufacture related products for improving skin aging or overall skin condition. This prior art does not specify the actual active component within the small collagen peptide molecules, nor does it explain the effects following the intervention from the perspective of natural aging. The invention and its advantages

[0005] To overcome the problems and shortcomings of the prior art, the present invention offers the use of an active peptide in the manufacture of a product for improving the skin barrier and promoting collagen expression and / or increasing skin elasticity and suppleness and / or reducing wrinkles. The present invention demonstrates that supplementation with exogenous precursors such as active peptides can improve collagen expression and the condition of the skin, while simultaneously increasing skin elasticity and suppleness and reducing wrinkles, and therefore has broad applications in areas such as dietary supplements, pet food, and others.When using the active peptide according to the present invention in the manufacture of a product for improving the skin barrier and promoting collagen expression, the products in question comprise at least one from general foods, functional foods, pet food and pharmaceuticals, thereby meeting the demand in various market segments.

[0006] According to a first aspect of the present invention, the use of an active peptide in the manufacture of a product for improving the skin barrier and promoting collagen expression and / or increasing skin elasticity and suppleness and / or reducing wrinkles is provided, wherein the active peptide has the structural formula HA-OH, A comprises at least two amino acid residues and the amino acid residues comprise at least one of Hyp (hydroxyproline), Gly (glycine), Pro (proline) and Ala (alanine).

[0007] The skin, as the largest organ of the human body, is the first to experience the effects of aging. Skin aging leads to a thinning of the epidermis and dermis, damaging the skin barrier (physical barrier, immune barrier, etc.). The basement membrane is a key connecting structure between the dermis and epidermis, and its health is crucial for normal metabolism and the skin's barrier function. During the skin aging process, for example, the expression of collagen XVII in the basement membrane decreases. Supplementing with components of the extracellular matrix, such as active peptides, can enhance the regeneration, differentiation, and migration of basement membrane cells, which is essential for repairing the skin barrier.

[0008] Furthermore, during the skin aging process, oxidative stress can activate matrix metalloproteinases, accelerate collagen degradation, and inhibit its regeneration, manifesting as skin photoaging, weakened barrier function, and pigmentary abnormalities. Oxidative stress and AGEs have a mutually reinforcing effect, forming a vicious cycle that accelerates aging. Simultaneously, cellular clearance capacity decreases during cell aging, leading to the persistent accumulation of oxidative metabolic end products such as lipofuscin within cells. Lipofuscin is difficult to degrade, can impair the function of lysosomes and mitochondria, further increase oxidative stress levels, and induce chronic inflammatory responses. It is closely associated with dull skin tone, blemishes, and reduced tissue repair capacity, and is a key feature of structural and functional skin aging.

[0009] Active peptide components are precursors of collagen in the human body. The present invention demonstrates that supplementation with exogenous precursors can improve collagen expression and the condition of the skin. In particular, it has been confirmed that the aforementioned active peptides effectively promote the expression of collagens such as COL12 and COL19 and can also reduce the degree of skin barrier damage, specifically manifested in a reduced expression of NLP-29. Therefore, the use of these active peptides can effectively improve the condition of the skin during natural aging, resulting in smoother, more translucent, and healthier skin.Furthermore, the present invention demonstrates that active peptides can strengthen the antioxidant capacity of the organism, reduce oxidative damage, lower the glycation level and the lipofuscin level in the organism, optimize the antioxidant defense system and thus delay the aging process by improving the enzyme activities of SOD and CAT in the organism.

[0010] Furthermore, confirmatory experiments of the present invention were conducted using the model organism Caenorhabditis elegans to verify the efficacy of the aforementioned active peptides. Collagen is the main component of the extracellular matrix (ECM). The extracellular matrix of nematodes is primarily distributed in the cuticle and basal lamina. Caenorhabditis elegans is a classic model organism for aging research. Due to its short life cycle and well-defined genetic background, the use of C. elegans as a model organism to investigate the effects of natural aging on the skin barrier represents a significant innovation. Caenorhabditis elegans offers advantages such as a well-defined genetic background, simple individual structure, short life cycle, a fully sequenced genome, and no ethical concerns. It is widely used in the fields of genetics and developmental biology, aging, and longevity.The skin barrier is considered an important indicator of aging, and the epidermis of Caenorhabditis elegans exhibits a high degree of homology to human skin. Therefore, Caenorhabditis elegans was chosen as a model organism for our functional validation.

[0011] Furthermore, the present invention uses naturally aging Caenorhabditis elegans as a model organism. It utilizes, for the first time, a natural skin barrier model of aging nematodes to demonstrate that the aforementioned active peptides can improve the skin barrier and skin conditions such as epidermal wrinkling under natural aging conditions. Moreover, it has been confirmed that the active peptides exhibit significant antioxidant and antiglycative effects, as well as the ability to reduce lipofuscin. This can offer new strategies for preventing damaged skin barriers in clinical skin aging, increasing skin elasticity and suppleness in aging skin, and improving wrinkles, and shows promising application prospects.At the same time, the present invention is also the first to use the aforementioned active peptides to improve damaged skin barriers under natural aging conditions, thereby expanding the application of products with the corresponding components in naturally aging populations. Furthermore, the present invention is also the first to use the natural aging of nematodes for skin-related intervention studies, thus expanding the application of nematodes in the field of skin aesthetics.

[0012] Furthermore, the present invention has also confirmed that the aforementioned active peptide molecules can increase skin density, enlarge the specific surface area of ​​the skin's basal membrane, increase the water content of the stratum corneum, improve skin elasticity, and reduce crow's feet. Therefore, these active peptide molecules can efficiently promote the production of the extracellular matrix in dermal cells, increase the dermis's support and water retention capacity, and boost the skin's metabolism and repair capacity, ultimately improving skin hydration, elasticity, and reducing wrinkles.

[0013] Preferably, the product for improving the skin barrier and promoting collagen expression includes at least one of H-Hyp-Gly-OH (OG), H-Gly-Pro-Ala-OH (GPA), H-Gly-Pro-Hyp-OH (GPO), and H-Gly-Pro-Hyp-Gly-OH (GPOG). The individual or combined effect of the aforementioned collagen-active peptides has been confirmed by relevant experiments, demonstrating their ability to effectively improve the skin barrier and promote collagen expression. This means that under natural aging conditions, they effectively improve skin condition, delay skin aging, and offer promising applications for interventions in natural aging.

[0014] Preferably, the product for increasing skin elasticity and suppleness and / or reducing wrinkles comprises at least one of H-Gly-Pro-Ala-OH (GPA) and H-Gly-Pro-Hyp-Gly-OH (GPOG). The two aforementioned active peptide molecules, used either individually, together, or in combination with other active peptide molecules, can effectively increase skin elasticity and suppleness and improve wrinkles, particularly through their antioxidant, antiglycation, and lipofuscin-reducing mechanisms, which increase skin hydration and elasticity and reduce wrinkles.Preferably, the promotion of collagen expression includes at least one of the following: (1) promotion of the expression of collagen COL-12; (2) promotion of the expression of collagen COL-19; (3) promotion of the expression of EMB-9 collagen; (4) improvement of epidermal wrinkling; the improvement of the skin barrier includes: reduction of the expression of the antimicrobial peptide NLP-29; the increase in skin elasticity and suppleness and / or reduction of wrinkles includes at least one of the following: (1) increase in SOD activity; (2) increase in CAT activity; (3) reduction in MDA content; (4) reduction in AGEs content; (5) reduction in lipofuscin content; (6) increase in skin density; (7) increase in the specific surface area of ​​the skin's basement membrane; (8) increase in the water content of the stratum corneum; (9) increase in skin elasticity; (10) improvement of skin and / or perianal wrinkles.Relevant experiments have verified that the present invention confirms that the aforementioned multiple active peptides can promote the expression of collagens COL-12, COL-19, and EMB-9, as well as decrease the expression of the antimicrobial peptide NLP-29. Therefore, they can effectively improve the skin barrier damaged by natural aging and reduce wrinkles, thus enhancing skin condition. Simultaneously, by increasing antioxidant and antiglycation levels and reducing lipofuscin, these multiple active peptide molecules can increase skin hydration and elasticity, thereby reducing wrinkles.

[0015] Preferably, the application goals of the product include improving the skin barrier and promoting collagen expression and / or increasing skin elasticity and suppleness and / or reducing wrinkles in animals and humans.

[0016] The product is preferably used to improve the skin barrier and promote collagen expression and / or increase skin elasticity and suppleness and / or reduce wrinkles, to improve the skin barrier damaged by natural aging in animals or humans and to promote collagen expression in humans or animals undergoing natural aging; and / or to increase skin elasticity and suppleness in animals or humans undergoing natural aging; and / or to improve wrinkles that develop in animals or humans undergoing natural aging.

[0017] Preferably includes the improvement of the skin barrier and promotion of collagen expression: H-Hyp-Gly-OH and / or H-Gly-Pro-Ala-OH and / or H-Gly-Pro-Hyp-OH and / or H-Gly-Pro-Hyp-Gly-OH to increase the expression of collagen COL-12 in middle-aged animals or humans.

[0018] Preferably, the product for increasing skin elasticity and suppleness and / or reducing wrinkles comprises at least one of H-Gly-Pro-Ala-OH and H-Gly-Pro-Hyp-Gly-OH.

[0019] Preferably, the improvement of the skin barrier and promotion of collagen expression includes: H-Gly-Pro-Hyp-OH and / or H-Gly-Pro-Hyp-Gly-OH to increase the expression of collagen COL-12 in animals or elderly humans.

[0020] Preferably includes the improvement of the skin barrier and promotion of collagen expression: H-Hyp-Gly-OH and / or H-Gly-Pro-Ala-OH and / or H-Gly-Pro-Hyp-OH and / or H-Gly-Pro-Hyp-Gly-OH to increase the expression of collagen COL-19 in young animals or humans.

[0021] Preferably includes the improvement of the skin barrier and promotion of collagen expression: H-Hyp-Gly-OH and / or H-Gly-Pro-Ala-OH and / or H-Gly-Pro-Hyp-OH and / or H-Gly-Pro-Hyp-Gly-OH to increase the expression of collagen COL-19 in middle-aged animals or humans.

[0022] Preferably, the improvement of the skin barrier and promotion of collagen expression includes: H-Gly-Pro-Hyp-OH and / or H-Gly-Pro-Hyp-Gly-OH to increase the expression of collagen COL-19 in elderly animals or humans.

[0023] Preferably includes the improvement of the skin barrier and promotion of collagen expression: H-Hyp-Gly-OH and / or H-Gly-Pro-Ala-OH and / or H-Gly-Pro-Hyp-OH and / or H-Gly-Pro-Hyp-Gly-OH to reduce the expression of the antimicrobial peptide NLP-29 in middle-aged animals or humans.

[0024] Preferably includes the improvement of the skin barrier and promotion of collagen expression: H-Hyp-Gly-OH and / or H-Gly-Pro-Ala-OH and / or H-Gly-Pro-Hyp-OH and / or H-Gly-Pro-Hyp-Gly-OH to improve epidermal wrinkling in middle-aged animals or humans.

[0025] Preferably includes the improvement of the skin barrier and promotion of collagen expression: H-Hyp-Gly-OH and / or H-Gly-Pro-Hyp-OH and / or H-Gly-Pro-Hyp-Gly-OH to improve epidermal wrinkling in animals or elderly humans.

[0026] Preferably includes the improvement of the skin barrier and promotion of collagen expression: H-Gly-Pro-Hyp-Gly-OH to increase the expression of EMB-9 collagen of the basement membrane in young animals or humans.

[0027] Preferably includes the improvement of the skin barrier and promotion of collagen expression: H-Gly-Pro-Ala-OH and / or H-Gly-Pro-Hyp-Gly-OH to increase the expression of EMB-9 basement membrane collagen in middle-aged animals or humans.

[0028] Preferably, the improvement of the skin barrier and promotion of collagen expression includes: H-Hyp-Gly-OH and / or H-Gly-Pro-Hyp-OH to increase the expression of EMB-9 collagen of the basement membrane in animals or elderly humans.

[0029] Preferably, the product for improving the skin barrier and promoting collagen expression comprises at least two of H-Hyp-Gly-OH, H-Gly-Pro-Ala-OH, H-Gly-Pro-Hyp-OH and H-Gly-Pro-Hyp-Gly-OH; or the product comprises at least three of H-Hyp-Gly-OH, H-Gly-Pro-Ala-OH, H-Gly-Pro-Hyp-OH and H-Gly-Pro-Hyp-Gly-OH; or the product comprises H-Hyp-Gly-OH, H-Gly-Pro-Ala-OH, H-Gly-Pro-Hyp-OH and H-Gly-Pro-Hyp-Gly-OH.

[0030] Preferably, the product for increasing skin elasticity and suppleness and / or reducing wrinkles comprises H-Gly-Pro-Ala-OH; or the product comprises H-Gly-Pro-Hyp-Gly-OH; or the product comprises H-Gly-Pro-Ala-OH and H-Gly-Pro-Hyp-Gly-OH; or the product comprises H-Gly-Pro-Ala-OH, H-Gly-Pro-Hyp-Gly-OH, H-Hyp-Gly-OH, H-Hyp-Pro-OH (OP) and H-Gly-Pro-Hyp-OH.

[0031] Preferably, the product for improving the skin barrier and promoting collagen expression comprises H-Hyp-Gly-OH and H-Gly-Pro-Ala-OH; or the product comprises H-Hyp-Gly-OH, H-Gly-Pro-Ala-OH and H-Gly-Pro-Hyp-OH; or the product comprises H-Hyp-Gly-OH, H-Gly-Pro-Hyp-OH and H-Gly-Pro-Hyp-OH; or the product comprises H-Hyp-Gly-OH, H-Gly-Pro-Ala-OH, H-Gly-Pro-Hyp-OH and H-Gly-Pro-Hyp-OH; or the product comprises H-Hyp-Gly-OH, H-Gly-Pro-Ala-OH, H-Gly-Pro-Hyp-OH, H-Gly-Pro-Hyp-OH and N-acetylglucosamine.

[0032] Preferably, if the product for improving the skin barrier and promoting collagen expression comprises H-Hyp-Gly-OH and H-Gly-Pro-Ala-OH, the mass ratio of H-Hyp-Gly-OH to H-Gly-Pro-Ala-OH is 4 to 6:4 to 6; or, if the product for improving the skin barrier and promoting collagen expression comprises H-Hyp-Gly-OH, H-Gly-Pro-Ala-OH and H-Gly-Pro-Hyp-OH, the mass ratio of H-Hyp-Gly-OH to H-Gly-Pro-Ala-OH to H-Gly-Pro-Hyp-OH is 0.1 to 3:4 to 9.8:0.1 to 3; or, if the product for improving the skin barrier and promoting collagen expression includes H-Hyp-Gly-OH, H-Gly-Pro-Hyp-OH and H-Gly-Pro-Hyp-Gly-OH, the mass ratio of H-Hyp-Gly-OH to H-Gly-Pro-Hyp-OH to H-Gly-Pro-Hyp-Gly-OH is 0.1 to 3: 4 to 9.8: 0.1 to 3;or, if the product for improving the skin barrier and promoting collagen expression comprises H-Hyp-Gly-OH, H-Gly-Pro-Ala-OH, H-Gly-Pro-Hyp-OH and H-Gly-Pro-Hyp-Gly-OH, the mass ratio of H-Hyp-Gly-OH to H-Gly-Pro-Ala-OH to H-Gly-Pro-Hyp-OH to H-Gly-Pro-Hyp-Gly-OH is 0.1 to 2: 2 to 3.9: 4 to 5.9: 0.1 to 2.;

[0033] According to a second aspect of the present invention, the use of an active peptide in the manufacture of a general food or functional food or pet food or drug is provided for improving the skin barrier and promoting collagen expression and / or increasing skin elasticity and suppleness and / or reducing wrinkles.

[0034] Preferably, the product for improving the skin barrier and promoting collagen expression and / or increasing skin elasticity and suppleness and / or reducing wrinkles comprises an oral preparation, wherein the oral preparation comprises at least one of the following: tablets, capsules, granules, pastes, liquid preparations, pills and suspensions.

[0035] According to a third aspect of the present invention, the use of an active peptide in the manufacture of a powder or gumball or beverage is provided to improve the skin barrier and promote collagen expression and / or increase skin elasticity and suppleness and / or reduce wrinkles.

[0036] Preferably, the active peptide in the form of a protein peptide is added to the product to improve the skin barrier and promote collagen expression and / or increase skin elasticity and suppleness and / or reduce wrinkles, wherein the mass fraction of the active peptide in the protein peptide is ≥5% and the amount of protein peptide added is 1 mg to 5000 mg per bottle, unit, or dose. Preferably, the number-mean molecular weight of the protein peptide is 250 to 500 Da. More preferably, the number-mean molecular weight of the protein peptide is 300 Da.

[0037] According to a fourth aspect of the present invention, the use of an active peptide and an amino sugar in the manufacture of a product for improving the skin barrier and promoting collagen expression and / or increasing skin elasticity and suppleness and / or reducing wrinkles is provided.

[0038] Preferably, the active peptide has the structural formula HA-OH, where A comprises at least two amino acid residues, and the amino acid residues include at least one from Hyp, Gly, Pro, and Ala; the amino sugar comprises N-acetylglucosamine. The combined use of the active peptide and the amino sugar can also promote the expression of collagens COL-12, COL-19, and EMB-9, as well as decrease the expression of the antimicrobial peptide NLP-29, while effectively improving the skin barrier damaged by natural aging and reducing wrinkles.

[0039] Preferably, when using the active peptide and the amino sugar in the manufacture of a product to improve the skin barrier and promote collagen expression and / or increase skin elasticity and suppleness and / or reduce wrinkles, the product comprises H-Hyp-Gly-OH, H-Gly-Pro-Ala-OH, H-Gly-Pro-Hyp-OH, H-Gly-Pro-Hyp-Gly-OH and N-acetylglucosamine (NAG).

[0040] Preferably, the mass ratio of H-Hyp-Gly-OH, H-Gly-Pro-Ala-OH, H-Gly-Pro-Hyp-OH, H-Gly-Pro-Hyp-Gly-OH and N-acetylglucosamine is 0.2 to 0.7: 3 to 3.5: 4.6 to 5.2: 0.2 to 0.7: 0.3 to 2.

[0041] In an optional embodiment, N-acetylglucosamine is used to manufacture a product for improving the skin barrier and promoting collagen expression, wherein the improvement of the skin barrier and promotion of collagen expression comprises: N-acetylglucosamine to increase the expression of collagen COL-12 in middle-aged animals or humans.

[0042] In an optional embodiment, N-acetylglucosamine is used to manufacture a product for improving the skin barrier and promoting collagen expression, wherein the improvement of the skin barrier and promotion of collagen expression comprises: N-acetylglucosamine to increase the expression of collagen COL-19 in young animals or humans.

[0043] In an optional embodiment, N-acetylglucosamine is used to manufacture a product for improving the skin barrier and promoting collagen expression, wherein the improvement of the skin barrier and promotion of collagen expression comprises: N-acetylglucosamine to reduce the expression of the antimicrobial peptide NLP-29 in young animals or humans.

[0044] In an optional embodiment, N-acetylglucosamine is used to manufacture a product for improving the skin barrier and promoting collagen expression, wherein the improvement of the skin barrier and promotion of collagen expression comprises: N-acetylglucosamine for improving epidermal wrinkling in middle-aged animals or humans.

[0045] In an optional embodiment, N-acetylglucosamine is used to manufacture a product for improving the skin barrier and promoting collagen expression, wherein the improvement of the skin barrier and promotion of collagen expression comprises: N-acetylglucosamine for improving epidermal wrinkling in animals or elderly humans.

[0046] According to a fifth aspect of the present invention, the use of an active peptide and an amino sugar in the manufacture of a general food or functional food or pet food or drug is provided for improving the skin barrier and promoting collagen expression and / or increasing skin elasticity and suppleness and / or reducing wrinkles.

[0047] According to a sixth aspect of the present invention, the use of an active peptide and an amino sugar in the manufacture of a powder or gumball or beverage is provided for improving the skin barrier and promoting collagen expression and / or increasing skin elasticity and suppleness and / or reducing wrinkles. Brief description of the drawings Fig. 1 and Fig. Figures 2 show the expression of epidermal collagen type I COL-12 in nematodes (at day 7) and nematodes (at day 15) in Example 1. Fig. 3, Fig. 4 and Fig. Figures 5 show the expression of epidermal collagen type II COL-19 in nematodes (at day 2), nematodes (at day 7) and nematodes (at day 15) in Example 2. Fig. 6 and Fig.Figures 7 show the expression of NLP-29 in nematodes (at day 2) and nematodes (at day 7) in Example 3. Fig. 8 and Fig. Figure 9 shows the REM images of the epidermis of nematodes (at day 7) and nematodes (at day 15) in Example 4. Fig. 10, Fig. 11 and Fig. Figures 12 show the expression of EMB-9 in nematodes (at day 2) and nematodes (at day 15) in Example 5. Fig. Figure 13 shows the influence of the active peptide combination on the rate of change of skin density. Fig. Figure 14 shows the influence of the active peptide combination on the rate of change of the specific surface area of ​​the basement membrane. Fig. Figure 15 shows the influence of the active peptide combination on the rate of change of the water content of the stratum corneum of the skin. Fig.Figure 16 shows the influence of the active peptide combination on the rate of change of the overall elasticity of the skin. Fig. Figure 17 shows the influence of the active peptide combination on the rate of change of the volume of the canthus wrinkles. Example(s) of implementation

[0048] To facilitate the understanding of the concept of the present invention by those skilled in the art, the technical solutions in the embodiments of the present invention are clearly and completely described below. Obviously, the described embodiments represent only a subset of the embodiments of the present invention and not all of them.

[0049] COL-12 is a collagen in the middle layer of the nematode cuticle. Its levels decline sharply in early adulthood, and it primarily serves a mechanical support function. COL-19 is a collagen found in the rings and lateral regions of the nematode cuticle, expressed in the outermost epidermis. Its mRNA expression declines sharply in early adulthood, but its protein level remains constant. The epidermal cell layer, as a physical barrier, is frequently exposed to various physical damages that compromise its structure. The immune system of the epidermal cells recognizes this structural damage as a danger signal, triggering an immune response that leads to the expression and release of the antimicrobial peptide NLP-29. Increased expression of NLP-29 indicates damage to the physical skin barrier.The age of the nematodes on day 2 corresponds to approximately 25 to 30 years in humans (young adults), day 7 corresponds to approximately 40 to 50 years (middle age) and day 15 corresponds to approximately 60 to 70 years (older age) in humans.

[0050] Therefore, in the specific implementation of the present invention, the expression of COL-12, COL-19 and NLP-29 in different life stages of the nematodes is investigated under the influence of the active peptides (OG, GPA, GPO, GPOG) and N-acetylglucosamine (NAG) in order to verify the effect of the said active peptides and of N-acetylglucosamine (NAG) on improving the skin barrier and promoting collagen expression during the natural aging process of animals or humans. Example 1

[0051] The influence of OG, GPA, GPO, GPOG and NAG on the expression of COL-12 was investigated in each case.

[0052] OG, GPA, GPO, GPOG, and NAG were each dissolved in water to prepare 1 M solutions. These were diluted with OP50 (a nutrient-deficient E. coli strain) to obtain 100 mM sample solutions. The nematode lines COL-12::DsRed and COL::DsRed, in which the collagen of the middle layer is labeled with a red fluorescent marker (DsRed), were used. The collagen content in the middle layer was assessed by the intensity of the red fluorescence (the higher the content, the stronger the fluorescence). The nematodes were synchronized. After reaching the L4 stage (48 to 60 h), they were transferred to FUdR medium (to inhibit egg laying), which was defined as day 0. Culture was maintained at a constant temperature of 20°C. Sample solutions were fed on day 0, day 5, and day 13. Fluorescence imaging and evaluation of the nematodes were performed under a reflected light fluorescence microscope on day 2, day 7 and day 15.For each statistical analysis, no fewer than 15 nematodes were recorded.

[0053] The results of the fluorescence imaging and intensity statistics are in Fig. 1 and Fig. 2 shown. This shows Fig. 1 or Fig. 2. The expression of epidermal collagen type I COL-12 in nematodes at day 7 and day 15, respectively. The mock group served as an untreated control group to which none of the active substances OG, GPA, GPO, GPOG or NAG were added.

[0054] As from Fig. As can be seen in Figure 1, compared to the mock group, the groups OG, GPA, GPO, GPOG and NAG were able to promote the expression of collagen COL-12 in middle-aged nematodes and showed a very significant increase effect.

[0055] Fig.2 showed that, compared to the mock group, the GPO and GPOG groups promoted the expression of collagen COL-12 in old age nematodes and exhibited a significant increase effect. Example 2

[0056] The influence of OG, GPA, GPO, GPOG and NAG on the expression of COL-19 was investigated in each case.

[0057] OG, GPA, GPO, GPOG, and NAG were each dissolved in water to prepare 1 M solutions. These were diluted with OP50 (a nutrient-deficient E. coli strain) to obtain 100 mM sample solutions. The nematode line COL-19::GFP, in which the collagen of the middle layer is labeled with a green fluorescent marker (GFP), was used. The collagen content in the middle layer was assessed based on the intensity of the green fluorescence (the higher the content, the stronger the fluorescence). The nematodes were synchronized. After reaching the L4 stage (48 to 60 h), they were transferred to FUdR medium (to inhibit egg laying), which was defined as day 0. Culture was maintained at a constant temperature of 20°C. Sample solutions were fed on day 0, day 5, and day 13. Fluorescence imaging and evaluation of the nematodes were performed under a reflected light fluorescence microscope on day 2, day 7 and day 15.For each statistical analysis, no fewer than 15 nematodes were recorded.

[0058] The results of the fluorescence imaging and intensity statistics are in Fig. 3, Fig. 4 and Fig. 5 shown. Fig. 3, Fig. 4 or Fig. 5. The expression of epidermal collagen type II COL-19 in nematodes at day 2, day 7, and day 15. The mock group served as an untreated control group to which none of the active substances OG, GPA, GPO, GPOG, or NAG were added.

[0059] As from Fig. As can be seen in Figure 3, compared to the mock group, the groups OG, GPA, GPO, GPOG and NAG were able to promote the expression of collagen COL-19 in young nematodes and showed a very significant increase effect.

[0060] Fig.4 showed that, compared to the mock group, the groups OG, GPA, GPO and GPOG promoted the expression of collagen COL-19 in middle-aged nematodes and exhibited a significant increase effect.

[0061] Fig. 5 showed that, compared to the mock group, the GPO and GPOG groups promoted the expression of collagen COL-19 in old age nematodes and exhibited a significant increase effect. Example 3

[0062] The influence of OG, GPA, GPO, GPOG and NAG on the expression of NLP-19 was investigated in each case.

[0063] The antimicrobial peptide NLP-29 is an immune response product expressed in nematodes following epidermal damage. The more severe the epidermal damage, the higher the level of expression. Therefore, it can be used to assess the extent of epidermal damage.

[0064] OG, GPA, GPO, GPOG, and NAG were each dissolved in water to prepare 1 M solutions. These were diluted with OP50 (a nutrient-deficient E. coli strain) to obtain 100 mM sample solutions. The nematode line NLP-19::GFP, in which the collagen of the middle layer is labeled with a green fluorescent marker (GFP), was used. The extent of damage to the collagen in the middle layer was assessed by the intensity of the green fluorescence (the less damage, the lower the fluorescence intensity). The nematodes were synchronized. After reaching the L4 stage (48 to 60 h), they were transferred to FUdR medium (to inhibit egg laying), which was defined as day 0. Culture was maintained at a constant temperature of 20°C. The sample solutions were fed on day 0 and day 5. On day 2 and day 7, fluorescence imaging and evaluation of the nematodes were performed under a reflected light fluorescence microscope.For each statistical analysis, no fewer than 15 nematodes were recorded.

[0065] The results of the fluorescence imaging and intensity statistics are in Fig. 6, Fig. 7 and Fig. 8 is shown. This shows Fig. 6. The expression of NLP-29 in nematodes at day 2, and Fig. 6 only shows the Mock Group and the NAG Group. Fig. Figure 7 shows the expression of NLP-29 in nematodes at day 7. Fig. Figure 8 shows the expression of NLP-29 in nematodes at day 15. The mock group served as an untreated control group to which none of the active substances OG, GPA, GPO, GPOG or NAG were added.

[0066] As from Fig. As can be seen in Figure 6, the NAG group showed a clear protective effect on the epidermis of young nematodes compared to the mock group.

[0067] Fig.7 showed that, compared to the mock group, the groups OG, GPA, GPO and GPOG all exhibited a significant protective effect on the epidermis of middle-aged nematodes. Example 4

[0068] The influence of OG, GPA, GPO, GPOG and NAG on the epidermal condition was investigated in each case.

[0069] OG, GPA, GPO, GPOG, and NAG were each dissolved in water to prepare 1 M solutions. These were diluted with OP50 (a nutrient-deficient E. coli strain) to obtain 100 mM sample solutions. The wild-type nematode strain N2 was synchronized. After the nematodes reached the L4 stage (48 to 60 h), they were transferred to FUdR medium (to inhibit egg laying), which was defined as day 0. Culture was maintained at a constant temperature of 20°C. The sample solutions were fed continuously. Scanning electron microscope (SEM) images were taken on days 7 and 15 for observation. No fewer than 5 nematodes were captured for each image series. The scanning electron microscope allowed direct visualization of the nematodes' epidermal condition.

[0070] The SEM image results are in Fig. 8 and Fig. 9 is shown. This shows Fig. 8 or Fig.9 SEM images of the epidermis of nematodes at day 7 and day 15. The mock group served as an untreated control group to which none of the active substances OG, GPA, GPO, GPOG or NAG were added.

[0071] As from Fig. As can be seen in Figure 8, the groups OG, GPA, GPO, GPOG and NAG showed a very significant improvement effect on the epidermis of middle-aged nematodes.

[0072] Fig. 9 showed that the groups OG, GPO, GPOG and NAG had a very significant improvement effect on the epidermis of nematodes in old age. Example 5

[0073] The influence of OG, GPA, GPO, GPOG and NAG on the expression of basement membrane collagen EMB-9 was investigated in each case.

[0074] EMB-9 collagen is a structural component of the extracellular matrix with tensile strength. It is involved in several processes, including the positive regulation of axon regeneration, the positive regulation of protein expression, and the serotonin response. It is localized in the basement membrane and expressed in various structures, including peritoneal cells, the ovulatory system, the gonad, the nerve ring, and rectal muscles. It is a homolog of human COL4A1 (collagen type IV alpha-1 chain). To observe this collagen type, a nematode strain was engineered with a green fluorescent protein-collagen fusion: emb-9::NLG. The stronger the green fluorescence, the more of this collagen is present.

[0075] First, OG, GPA, GPO, GPOG, and NAG were each dissolved in water to prepare 1 M solutions. These were diluted with OP50 (a nutrient-deficient E. coli strain) to obtain 100 mM sample solutions. The emb-9::NLG nematode strain was synchronized. After the nematodes reached the L4 stage (48 to 60 h), they were transferred to FUdR medium, which was defined as day 0. Culture was maintained at a constant temperature of 20°C. On days 0, 5, and 13, the sample solutions were fed, and the nematodes were cultured for two days. On days 2, 7, and 15, images and analyses of the nematodes were performed using a reflected-light fluorescence microscope. At least 15 nematodes were recorded for each image series.

[0076] The results of the fluorescence imaging and intensity statistics are in Fig. 10, Fig. 11 and Fig. 12 is shown. This shows Fig.10 the expression of EMB-9 in nematodes at day 2, and Fig. 10 only includes the Mock Group, the GPOG Group and the NAG Group. Fig. Figure 11 shows the expression of EMB-9 in nematodes at day 7, and Fig. Item 11 only shows the mock, GPA, and GPOG groups. Fig. Figure 12 shows the expression of EMB-9 in nematodes at day 15, and Fig. Sample 12 contains only the mock, OG, GPO, and NAG groups. The mock group served as an untreated control group to which none of the active substances OG, GPA, GPO, GPOG, or NAG were added.

[0077] As from Fig. As can be seen in section 10, the GPOG group and the NAG group showed a significant protective effect on the epidermis of young nematodes compared to the mock group.

[0078] Fig.11 showed that, compared to the mock group, the GPA group and the GPOG group both exhibited a significant protective effect on the epidermis of middle-aged nematodes.

[0079] Fig. 12 showed that, compared to the mock group, the OG group, the GPO group and the NAG group exhibited a significant protective effect on the epidermis of nematodes in old age. Example 6

[0080] The expression of COL-12, NLP-29 and EMB-9 was investigated for the combinations OG+GPOG, OG+GPA+GPOG, OG+GPO+GPOG, OG+GPA+GPO+GPOG and OG+GPA+GPO+GPOG+NAG.

[0081] Samples of the combinations OG+GPOG, OG+GPA+GPOG, OG+GPO+GPOG, OG+GPA+GPO+GPOG, and OG+GPA+GPO+GPOG+NAG were dissolved in water according to the mass ratio combinations specified in Table 1 to prepare 1 M solutions. These were diluted with OP50 (a nutrient-deficient E. coli strain) to obtain 100 mM sample solutions. The wild-type nematode strain N2 was synchronized. After the nematodes reached the L4 stage (48 to 60 h), they were transferred to FUdR medium (to inhibit egg laying), which was defined as day 0. Culture was maintained at a constant temperature of 20°C. Sample solutions were fed on day 0, day 5, and day 13. On days 2, 7, and 15, the parameters COL-12 and NLP-29 were detected using a fluorescence microscope. The relevant results are also shown in Table 1.The data in the columns for COL-12 (Day 7), COL-12 (Day 15), NLP-29 (Day 2), and NLP-29 (Day 7) are based on calculations relative to the mock group. The mock group served as an untreated control group to which none of the active substances OG, GPA, GPO, GPOG, or NAG were added. The corresponding symbol “ / ” indicates that there was no significant effect compared to the mock group; that is, it was similar to or even worse than the mock group.

[0082] Table 1 Expression of collagen for the different compositions in Example 6 project Ratio (mass ratio) COL-12 (Day 7) COL-12 (Day 15) NLP-29 (Day 2) NLP-29 (Day 7) EMB-9 (Day 2) EMB-9 (Day 7) EMB-9 (Day 15) OG:GPOG 6: 4 +33,4% +12,1% / 35,2% / +10,3% +12,3% 5:5 +40,2% +15,2% / - 41,3 % / +13,4% +12,2% 4: 6 +34,3% +13,1% / 32,3% / +12,1% +12,3% OG: GPA:GPOG 3: 4: 3 +25,2% +11,2% / 32,1 / +50,3% +12,5% % 0,5: 9:0,5 +36,2% +16,8% / 37,6% / +53,2% +13,4% 0,1: 9,8:0,1 +26,2% +10,1% / 30,2% / +52,1 % +14,4% OG: GPO:GPOG 3:4:3 +28,3% +17,7% / 18,4% / +10,2% +24,1 % 0,3: 9,4:0,3 +32,1% +25,2% / 25,4% / +12,1% +23,2% 0,1: 9,8:0,1 +26,3% +16,53% / 19,5% / +11,2% +21,5% OG: GPA:GPO:GPOG 2: 2: 4:2 +32,3% +19,4% / 18,3% +12,4% +34,5% +23,2% 0,2: 3,8:5,8: 0,2 +43,2% +26,3% / - 21,6 % +15,2% +35,2% +23,5% 0,1: 3,9:5,9: 0,1 +34,1% +19,7% / 17,5% +13,3% +33,1% +22,4% OG: GPA:GPO:GPOG:NAG 0,7: 3:4,6: 0,7:1 +32,3% / - 38,2 % - 23,5 % +19,8% +32,2% +40,1 % 0,2: 3:4,6: 0,2:2 +46,3% / - 40,5 % - 30,6 % +22,2% +33,2% +41,2% 0,2: 3,5:5,2: 0,2:0,9 +45,7% / - 42,3 % - 28,7 % +20,1% +34,3% +39,5% 0,5: 3,5:5,2: 0,5:0,3 +33,0% / - 37,5 % - 24,3 % +18,3% +34,2% +98,9%

[0083] Table 1 shows that OG+GPOG, OG+GPA+GPOG, OG+GPO+GPOG, OG+GPA+GPO+GPOG and OG+GPA+GPO+GPOG+NAG significantly improve collagen expression and epidermal barrier function in nematodes under natural aging conditions in specific ratio ranges. Example 7

[0084] The influence of GPA and GPOG on SOD activity, CAT activity, MDA content, AGEs levels and lipofuscin levels was investigated in each case.

[0085] SOD is superoxide dismutase, which specifically eliminates the most common and initial harmful free radical, the superoxide anion radical, produced during energy metabolism in the body. SOD can convert it into the less toxic hydrogen peroxide. Increased SOD activity means that the body's ability to eliminate initial free radicals is significantly enhanced. This is a key indicator that the antioxidant defense system has been activated or protected.

[0086] CAT is catalase, which specifically breaks down the hydrogen peroxide produced by the SOD-catalyzed reaction, converting it into completely harmless water and oxygen. When hydrogen peroxide accumulates, more destructive free radicals can be formed under the catalysis of metal ions. Increased CAT activity indicates that the organism's ability to efficiently process hydrogen peroxide and prevent secondary damage is significantly enhanced. Together with SOD, it forms a complete enzymatic antioxidant defense line.

[0087] MDA is malondialdehyde, the end product of lipid peroxidation and the "debris" or "rust" left behind after oxidative damage to cells. When free radicals attack the unsaturated fatty acids in the cell membrane, they trigger a chain reaction that leads to lipid peroxidation. MDA is the stable end metabolite of this process. Its level directly reflects the severity of oxidative damage to the cell membrane.

[0088] Advanced glycation end products (AGEs) are a class of irreversible, cross-linking, and fluorescent harmful compounds formed by non-enzymatic reactions of proteins or fats with sugar molecules in the body. With increasing age and rising blood sugar levels, AGEs accumulate in tissues. AGE accumulation disrupts normal protein function, promotes inflammation and oxidative stress, and is a key factor in skin aging (wrinkles, loss of elasticity), atherosclerosis, diabetes complications, and neurodegenerative diseases. It is the core molecule that links "glycation" and "aging."

[0089] Lipofuscin, colloquially known as "age pigment," is an inert fluorescent substance formed by the accumulation of incompletely degraded protein and lipid residues (including oxidatively damaged mitochondria, cell organelles, etc.) in lysosomes following autophagy. Lipofuscin accumulation is a marker for cellular senescence and loss of function, particularly in nerve cells and cardiac muscle cells. It occupies cellular space, disrupts normal function, and its concentration directly reflects the efficiency of the intracellular waste disposal system and the extent of long-term oxidative damage accumulation.

[0090] The specific experimental methodology for investigating the influence of GPA and GPOG on the above-mentioned parameters was as follows: (1) Nematode culture: The cultured E. coli OP50 was dripped onto the prepared NGM medium and evenly spread with a streaking stick, with the bacterial coating approximately 5 mm from the edge of the culture dish. The coated NGM plates were dried and incubated for 24 h at 37°C in an incubator until the colonies completely covered the plate and were ready for use. In the sterile workbench, one-quarter of an NGM plate containing L4 or adult nematodes was cut out and transferred to a new NGM plate. The culture was carried out in a nematode incubator at a constant 20°C and 50% relative humidity. Passage was performed every 2 to 3 days, also using the plate cutout method, to maintain nematode viability and prevent contamination. (2) Nematode synchronization: Under the microscope, it was observed whether the nematodes had reached the L4 or adult stage for synchronization. The surface of the NGM medium was rinsed with M9 buffer to collect the nematodes. They were collected in a 1.5 mL centrifuge tube and centrifuged for 3 min at 1500 rpm. The supernatant was discarded. 1 mL of M9 buffer was added to wash the nematodes. Then, the mixture was centrifuged for 1 min at 1500 rpm, and the supernatant was discarded. This step was repeated three times. First, 0.5 mL of M9 buffer was added, followed by 0.5 mL of lysis solution, mixed, and shaken for 3 min. Then, the mixture was centrifuged for 3 min at 1500 rpm, and the supernatant was discarded. 1 mL of M9 buffer was added to wash the nematodes. The mixture was then centrifuged for 1 minute at 1500 rpm and the supernatant discarded. This step was repeated three times.An appropriate amount of M9 buffer was added, and the suspension was dropwise onto NGM medium coated with E. coli OP50. Culture was performed in an incubator. (3) Testing the oxidative, glycation and lipofuscin levels of the nematodes: a. Oxidation: The synchronized nematodes were transferred to the control group (without additive) and the sample groups. The concentrations of the two sample groups were GPA 300 µM and GPOG 500 µM, respectively. After 5 days of culture in the nematode incubator, the nematodes were removed, collected, and washed with M9 buffer. After centrifugation, the pellet was retained. CAT and SOD activity, as well as MDA content, were determined according to the instructions of the respective test kits. The nematode tissue pellet and the corresponding extraction solution were completely homogenized under cooling in a high-throughput cryomill. The homogenate was used for the measurements. b. Glycation: Glucose solution was added to the medium, resulting in a glucose concentration of 40 mmol / L in the agar. At this concentration, the glucose concentration in the total extract of C. elegans was 14 mmol / L. The glucose was applied to the plates one day prior to incubation. Synchronized nematodes were transferred to the control and test groups, respectively. The concentration of active peptides was 300 µM for the GPA group and 500 µM for the GPOG group. After 96 h of culture in the nematode incubator, the nematodes were removed, collected, and washed with M9 buffer. After centrifugation, the pellet was retained. The AGEs content was determined according to the instructions of the respective test kit. The nematode tissue pellet and the corresponding extraction solution were thoroughly homogenized under cooling in a high-throughput cryomill. The homogenate was used for measurement. c. Cellular senescence (assessed via lipofuscin levels in nematodes): Synchronized L4-stage C. elegans were transferred to NGM medium with the corresponding treatment groups (control group and two sample groups). The concentrations for GPA and GPOG were 300 µM each (sample groups). Transfer was performed daily for 10 days. On day 10, 10 worms aged 10 days were randomly selected from each group, anesthetized with a 1% tetramisohydrochloride solution, and placed on an agarose pad. Observation and imaging were performed using an inverted fluorescence microscope with excitation and emission wavelengths of 380 nm and 430 nm, respectively. Fluorescence intensity was quantitatively analyzed using ImageJ software.

[0091] The relevant test results are shown in Table 2, Table 3 and Table 4. Table 2 Test results on the antioxidant capacity of nematodes group Model group GPA Group GPOG Group SOD(U / mg.protein) 7,05 8,74 13,06 CAT(U / mg.prot) 12,25 30,61 19,48 MDA (nmol / mg) 19,52 15,93 5,77 Table 3 Test results for AGEs content in nematodes group Model group GPOG Group Improvement rate AGEs (ng / mL) 24,7 16,9 31,7% Table 4 Test results for lipofuscin in nematodes group Model group GPA Group GPOG Group Lipofuscin 82,00 59,18 63,01

[0092] Table 2 showed that the active peptides GPA and GPOG increased SOD activity in nematodes by 24% and 85%, respectively; increased CAT activity by 150% and 59%, respectively; and decreased MDA by 18% and 70%, respectively. This demonstrates that the active peptides GPA and GPOG enhance the organism's antioxidant capacity and reduce oxidative damage by improving the enzyme activities of SOD and CAT.

[0093] Table 3 showed that the active peptide GPOG reduced glycation end products (AGEs) levels in nematodes by 31.7%. This demonstrates that GPOG can lower glycation levels in the organism. Combined with the results of the antioxidant experiment, it is assumed that by increasing the activity of antioxidant enzymes, the organism reduces oxidative levels, thus achieving an antiglycative effect.

[0094] Table 4 showed that the active peptides GPA and GPOG reduced lipofuscin levels in nematodes by 28% and 23%, respectively. This indicates that the active peptides GPA and GPOG strengthen the antioxidant defense system of nematodes and slow down the aging process. Example 8

[0095] The influence of collagen peptides containing GPA+GPOG+OG+PO+GPO on increasing skin elasticity and suppleness, as well as improving wrinkles, was investigated. The proportion of GPO+GPA was ≥4.5% and the proportion of GPO+GPA+OG+PO+GPOG was ≥5%.

[0096] Nematode culture and synchronization were performed as in Example 7. Subsequently, the following steps were carried out to test the skin condition: Under normal conditions, adult subjects consumed the samples to be evaluated for 84 days according to the instructions in order to assess the effect of the two samples on the improvement of fine facial wrinkles, elasticity and firmness, hydration and regeneration, content and morphology of collagen and elastin fibers in the dermis, morphology of dermal cells, and dermal density, thickness, radiance, and smoothness of the skin.

[0097] High-dose group: A total of 35 individuals (all female, healthy, and of Chinese origin) successfully completed the assessment. Their ages ranged from 31 to 45 years, with a mean age of 39.06 ± 4.08 years. They met the inclusion and exclusion criteria for study volunteers.

[0098] Low-dose group: A total of 34 individuals (all female, healthy, and of Chinese origin) successfully completed the assessment. Their ages ranged from 31 to 45 years, with a mean age of 40.12 ± 3.91 years. They met the inclusion and exclusion criteria for study volunteers.

[0099] Control group: A total of 32 individuals (all female, healthy, and of Chinese origin) successfully completed the assessment. Their ages ranged from 31 to 45 years, with a mean age of 38.31 ± 4.45 years. They met the inclusion and exclusion criteria for study volunteers.

[0100] Sample information: The beverages were pasteurized acidic drinks. The high-dose group received 2.5 g of protein peptide / day, the low-dose group 1 g of protein peptide / day, and the control group 0 g of protein peptide / day.

[0101] Method of administration: Once daily, one bottle per dose, before bedtime.

[0102] Evaluation period: Before sample administration (D0), 14 days after sample administration (D14), 28 days after sample administration (D28), 56 days after sample administration (D56), 84 days after sample administration (D84).

[0103] Experimental material: The protein peptide was produced from collagen-rich fish scales, fish skin, bovine hide, or bovine bone as raw materials. Targeted enzymatic hydrolysis with specific proteases yielded a protein peptide with a number-mean molecular weight of approximately 300 Da, where the content of active peptides (OG+PO+GPO+GPA+GPOG) was ≥5% (by mass).

[0104] Upon arrival at the clinical trial center, participants cleansed their faces with a cleansing product and then sat for 30 minutes in a laboratory room at a temperature of 21 ± 2°C and 50 ± 10% relative humidity. Laboratory personnel acquired Primos CR images of the participants' faces, which were used to capture periareolar images and analyze fine wrinkles. The Corneometer® CM 825 skin measurement device was used to measure and analyze the water content of the stratum corneum. The Cutometer® dual MPA580 was used to measure and analyze skin elasticity and firmness. The Dermalab® Combo was used to measure and analyze facial skin density. A two-photon microscopy system was used to image and analyze the specific surface area of ​​the basal membrane.

[0105] The corresponding test results are in Fig. 13, Fig. 14, Fig. 15, Fig. 16 and Fig. 17 shown.

[0106] Fig. Study 13 showed that the rate of change in skin density compared to baseline gradually increased with increasing duration of administration, reaching 44.9% in the high-dose group after 12 weeks. Collagen, elastin, and hyaluronic acid in the dermis decrease with age, leading to a reduction in skin density and consequently a diminished support and water retention capacity of the dermis. Oral administration of a collagen peptide containing the active peptides GPA, GPOG, OG, PO, and GPO at a dosage of 2.5 g / day increased skin density by 44.9%. This demonstrates that this collagen peptide, with its antioxidant, antiglycative, and lipofuscin-reducing properties, can efficiently promote the production of extracellular matrix in the dermis, contributing to skin support and water retention, thus improving skin hydration, elasticity, and wrinkles.

[0107] Fig.Study 14 showed that, compared to the baseline, the specific surface area of ​​the skin's basement membrane was larger after 12 weeks of supplementation, with an improvement of 16.5% in the low-dose group and 17.2% in the high-dose group. The basement membrane, the connecting layer between the dermis and epidermis of the skin, consists of components such as collagen and glycoproteins. It is responsible for the connection, nourishment, and communication between the dermis and epidermis and plays an important role in maintaining skin health, promoting repair and regeneration, and protecting against external irritants. With increasing age, the basement membrane becomes thinner and its specific surface area decreases. Oral administration of collagen peptides containing the active peptides GPA, GPOG, OG, PO, and GPO can significantly increase the specific surface area of ​​the basement membrane.This indicates that the “bridge” connecting the deep and superficial layers of skin is widening and communication between them is running more smoothly, which supports the metabolism and repair capacity of the skin.

[0108] Fig. 15 showed that the rate of change of the water content of the stratum corneum of the skin compared to the state before ingestion gradually increased with increasing duration of ingestion and reached 34.42% after 12 weeks in the high-dose group.

[0109] Fig. 16 showed that the rate of change of total skin elasticity R2 compared to the pre-intake state gradually increased with increasing duration of intake and reached 20.8% after 12 weeks in the high-dose group.

[0110] Fig.17 showed that the rate of change in the volume of the angle of the eye folds compared to the state before administration gradually increased with increasing duration of administration and reached 30.6% after 12 weeks in the high-dose group.

[0111] It should be further explained that orally ingested collagen peptides, if they only contain active peptides that promote the production of extracellular matrix such as collagen, elastin, and hyaluronic acid, do not address the problems of oxidative damage, glycation, and lipofuscin accumulation associated with skin aging. Oxidative stress and the formation of AGEs lead to the activation of matrix metalloproteinases, accelerating collagen degradation and inhibiting its regeneration, thus partially negating the effect of collagen supplementation.

[0112] Collagen peptides containing the active peptides GPA and GPOG, on the other hand, can activate the body's antioxidant system, reduce the formation of AGEs, decrease lipofuscin accumulation, and thus delay cellular aging. This allows the active peptides OG, PO, and GPO to produce collagen, elastin, and hyaluronic acid more efficiently. Simultaneously, the activation of matrix metalloproteinases is reduced, and the degradation of collagen and elastin is decreased. This promotes the production and maintenance of collagen and elastin in the dermis, increases skin density, and consequently boosts skin hydration and elasticity while reducing wrinkles.

[0113] Due to the antioxidant and antiglycation effects mediated by GPA and GPOG, as well as the delay of cellular aging through lipofuscin reduction, the extracellular matrix in dermal cells is more efficiently increased and accumulated. As a result, skin density increases by almost half (44.9%), contributing to skin support and water retention. The increase in the specific surface area of ​​the basal membrane indicates a smoother connection between the dermis and epidermis, which facilitates the supply of moisture and nutrients to the epidermis and the transport and elimination of metabolic waste products. Ultimately, this leads to a significant improvement in skin hydration, elasticity, and wrinkle reduction.

[0114] The examples above serve only to illustrate the technical solution of the present invention and not to limit the scope of protection of the invention. Although the invention has been described in detail with reference to the examples above, those skilled in the art should understand that the technical solution of the present invention can be modified or replaced with an equivalent solution. However, all such modifications or replacements fall within the scope of protection of the present invention.

Claims

[1] Use of an active peptide in the manufacture of a product to improve the skin barrier and promote collagen expression and / or increase skin elasticity and suppleness and / or reduce wrinkles, characterized by , that the active peptide has the structural formula HA-OH, wherein A comprises at least two amino acid residues, and the amino acid residues comprise at least one from Hyp, Gly, Pro and Ala. [2] Use of the active peptide according to claim 1 in the manufacture of a product for improving the skin barrier and promoting collagen expression and / or increasing skin elasticity and suppleness and / or reducing wrinkles, characterized by , that, If the product is used to improve the skin barrier and promote collagen expression, the product must contain at least one of H-Hyp-Gly-OH, H-Gly-Pro-Ala-OH, H-Gly-Pro-Hyp-OH, and H-Gly-Pro-Hyp-Gly-OH; and If the product is used to increase skin elasticity and suppleness and / or reduce wrinkles, the product must contain at least one of H-Gly-Pro-Ala-OH and H-Gly-Pro-Hyp-Gly-OH. [3] Use of the active peptide according to claim 1 in the manufacture of a product for improving the skin barrier and promoting collagen expression and / or increasing skin elasticity and suppleness and / or reducing wrinkles, characterized by , that The promotion of collagen expression includes at least one of the following: (1) promotion of the expression of collagen COL-12; (2) promotion of the expression of collagen COL-19; (3) promotion of the expression of EMB-9 collagen; (4) improvement of epidermal wrinkling; Improving the skin barrier includes: reducing the expression of the antimicrobial peptide NLP-29; The increase in skin elasticity and suppleness and / or reduction of wrinkles includes at least one of the following: (1) increase in SOD activity; (2) increase in CAT activity; (3) decrease in MDA content; (4) decrease in AGEs content; (5) decrease in lipofuscin content; (6) increase in skin density; (7) increase in the specific surface area of ​​the skin's basement membrane; (8) increase in the water content of the stratum corneum; (9) increase in skin elasticity; (10) improvement of skin and / or perianal wrinkles. [4] Use of the active peptide according to claim 1 in the manufacture of a product for improving the skin barrier and promoting collagen expression and / or increasing skin elasticity and suppleness and / or reducing wrinkles, characterized by that the application targets include animals and humans. [5] Use of the active peptide according to claim 1 in the manufacture of a product for improving the skin barrier and promoting collagen expression and / or increasing skin elasticity and suppleness and / or reducing wrinkles, characterized by , that it is used to improve the skin barrier in animals or humans undergoing natural aging and to promote collagen expression in humans or animals undergoing natural aging; and / or to increase skin elasticity and suppleness in animals or humans undergoing natural aging; and / or for improving wrinkles that develop in animals or humans through natural aging. [6] Use of the active peptide according to claim 3 in the manufacture of a product to improve the skin barrier and promote collagen expression and / or increase skin elasticity and suppleness and / or reduce wrinkles, characterized by, that the improvement of the skin barrier and promotion of collagen expression and / or increase in skin elasticity and suppleness and / or reduction of wrinkles includes at least one of the following: (1)H-Hyp-Gly-OH and / or H-Gly-Pro-Ala-OH and / or H-Gly-Pro-Hyp-OH and / or H-Gly-Pro-Hyp-Gly-OH is used to increase the expression of collagen COL-12 in middle-aged animals or humans; or H-Gly-Pro-Hyp-OH and / or H-Gly-Pro-Hyp-Gly-OH is used to increase the expression of collagen COL-12 in elderly animals or humans; (2)H-Hyp-Gly-OH and / or H-Gly-Pro-Ala-OH and / or H-Gly-Pro-Hyp-OH and / or H-Gly-Pro-Hyp-Gly-OH is used to increase the expression of collagen COL-19 in young animals or humans; or H-Hyp-Gly-OH and / or H-Gly-Pro-Ala-OH and / or H-Gly-Pro-Hyp-OH and / or H-Gly-Pro-Hyp-Gly-OH is used to increase the expression of collagen COL-19 in middle-aged animals or humans; or H-Gly-Pro-Hyp-OH and / or H-Gly-Pro-Hyp-Gly-OH is used to increase the expression of collagen COL-19 in elderly animals or humans; (3)H-Hyp-Gly-OH and / or H-Gly-Pro-Ala-OH and / or H-Gly-Pro-Hyp-OH and / or H-Gly-Pro-Hyp-Gly-OH is used to reduce the expression of the antimicrobial peptide NLP-29 in middle-aged animals or humans; (4)H-Hyp-Gly-OH and / or H-Gly-Pro-Ala-OH and / or H-Gly-Pro-Hyp-OH and / or H-Gly-Pro-Hyp-Gly-OH is used to improve epidermal wrinkling in middle-aged animals or humans; or H-Hyp-Gly-OH and / or H-Gly-Pro-Hyp-OH and / or H-Gly-Pro-Hyp-Gly-OH is used to improve epidermal wrinkling in elderly animals or humans; (5)H-Gly-Pro-Hyp-Gly-OH is used to increase the expression of EMB-9 collagen in young animals or humans; or H-Gly-Pro-Ala-OH and / or H-Gly-Pro-Hyp-Gly-OH is used to increase the expression of EMB-9 collagen in middle-aged animals or humans; or H-Hyp-Gly-OH and / or H-Gly-Pro-Hyp-OH is used to increase the expression of EMB-9 collagen in elderly animals or humans. [7] Use of the active peptide according to claim 1 in the manufacture of a product for improving the skin barrier and promoting collagen expression and / or increasing skin elasticity and suppleness and / or reducing wrinkles, characterized by , that the product comprises at least two of H-Hyp-Gly-OH, H-Gly-Pro-Ala-OH, H-Gly-Pro-Hyp-OH and H-Gly-Pro-Hyp-Gly-OH; or the product comprises at least three of H-Hyp-Gly-OH, H-Gly-Pro-Ala-OH, H-Gly-Pro-Hyp-OH and H-Gly-Pro-Hyp-Gly-OH; or the product includes H-Hyp-Gly-OH, H-Gly-Pro-Ala-OH, H-Gly-Pro-Hyp-OH and H-Gly-Pro-Hyp-Gly-OH. [8] Use of the active peptide according to claim 1 in the manufacture of a product for improving the skin barrier and promoting collagen expression and / or increasing skin elasticity and suppleness and / or reducing wrinkles, characterized by , that, if the product is used to improve the skin barrier and promote collagen expression, the product contains H-Hyp-Gly-OH and H-Gly-Pro-Ala-OH; or the product contains H-Hyp-Gly-OH, H-Gly-Pro-Ala-OH and H-Gly-Pro-Hyp-OH; or the product contains H-Hyp-Gly-OH, H-Gly-Pro-Hyp-OH and H-Gly-Pro-Hyp-OH; or the product contains H-Hyp-Gly-OH, H-Gly-Pro-Ala-OH, H-Gly-Pro-Hyp-OH and H-Gly-Pro-Hyp-OH; and if the product is used to increase skin elasticity and suppleness and / or reduce wrinkles, the product contains H-Gly-Pro-Ala-OH; or the product contains H-Gly-Pro-Hyp-Gly-OH; or the product contains H-Gly-Pro-Ala-OH and H-Gly-Pro-Hyp-Gly-OH; or the product contains H-Gly-Pro-Ala-OH, H-Gly-Pro-Hyp-Gly-OH, H-Hyp-Gly-OH, H-Hyp-Pro-OH (OP) and H-Gly-Pro-Hyp-OH. [9] Use of an active peptide and an amino sugar in the manufacture of a product to improve the skin barrier and promote collagen expression and / or increase skin elasticity and suppleness and / or reduce wrinkles, characterized by , that the active peptide has the structural formula HA-OH, wherein A comprises at least two amino acid residues, and the amino acid residues comprise at least one from Hyp, Gly, Pro and Ala; and The amino sugar N-acetylglucosamine is included. [10] Use of the active peptide according to any one of claims 1 to 8 in the manufacture of a product for improving the skin barrier and promoting collagen expression and / or increasing skin elasticity and suppleness and / or reducing wrinkles, or use of the active peptide and the amino sugar according to claim 9 in the manufacture of a product for improving the skin barrier and promoting collagen expression and / or increasing skin elasticity and suppleness and / or reducing wrinkles, characterized by that the product includes at least one of the following: general foodstuffs, functional foods, pet food and pharmaceuticals.

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