Recombinant XVII type collagen for skin aging resistance as well as preparation method and application of recombinant XVII type collagen

Abstract

The invention relates to an anti-aging oligopeptide for beauty and skin care, the oligopeptide has an amino acid sequence as shown in (X1) (R or K or H) G (D or E) # (X2), X1 represents one amino acid of G, A, V, L, I and P or represents that no amino acid exists, # represents one amino acid selected from A, V, D, E, N and Q, and X2 represents G and P or represents that no amino acid exists. The oligopeptide is a functional fragment of XVII type collagen, can induce in-vivo collagen synthesis or simulate a protein regulation function, has the effects of promoting cell self-cleaning, helping to remove aged skin cells, reconstructing skin rhythm, maintaining a good skin structure, resisting initial aging, promoting skin repair and regeneration, reducing fine wrinkles and the like, and is mild in action, free of side effects and free of toxic and side effects. The product has no obvious adverse reaction on human body, and is a good beautifying and anti-aging product.

Description

[0001] This case is a divisional application of the invention patent application with the application date of December 16, 2022, application number CN202211621908.5, and invention name "Recombinant type XVII collagen for skin anti-aging, its preparation method and application". Technical Field

[0002] The invention belongs to the field of cosmetics, and in particular relates to an oligopeptide with anti-aging effect and a use thereof. Background Art

[0003] In organisms, the failure of defense and repair mechanisms leads to the accumulation of damage, which in turn causes aging. In order to maintain homeostasis, cells effectively repair DNA damage and correct spontaneous replication errors, and even eliminate mutant cells in tissues. In this process, damaged or mutated senescent cells are cleared and replaced by adult stem cell differentiation competition to achieve the goal of cell self-purification. With age, the accumulation of senescent cells and the decline in the differentiation ability of adult stem cells lead to the aging of organs and even organisms.

[0004] The same is true for the skin, the largest organ in the human body. Damaged and aged cells in the young epidermis and dermis are constantly replaced by competition to maintain their function and morphology. The self-purification ability of cells in aged skin weakens, showing accumulation of senescent cells, reduction of functional melanocytes and dermal fibroblasts, and reduction of thickness. Therefore, the skin of the elderly becomes atrophic (thinning), fragile, loose, wrinkled, pigmented, and has delayed wound healing.

[0005] As a large family of proteins, collagen is often recognized as a mechanical support filler in mammals, but its function as a transmembrane signaling molecule is rarely explored. The epidermis is the outermost layer of the skin and can resist the invasion of foreign bodies and physical damage, playing a protective role. The basal cells in the innermost layer of the epidermis are in a state of continuous proliferation, which allows the epidermis to continuously achieve self-purification and regeneration. In this process, the epidermal growth factor receptor (EGFR) is required for signal transduction and cell migration regulation of type XVII collagen. The semi-demosomes formed by type XVII collagen connect epidermal stem cells to the basement membrane, while regulating the actin and keratin networks, coordinating the migration of epidermal stem cells to renew self-purifying epidermal cells. Therefore, type XVII collagen plays a very important regulatory role in regulating skin renewal and self-purification. Studies have shown that type XVII collagen (Col17a1) decreases in the elderly. In addition to the decline over time, the increasing pressure of life and work, environmental pollution, electronic radiation, staying up late, and the intake of high-calorie foods also accelerate the loss of type XVII collagen. Ultraviolet rays, radiation and oxidative stress in the environment can also cause the degradation and loss of type XVII collagen. In addition, Col17a1 human gene defects can also lead to bullous junctional epidermolysis bullosa, which presents with skin atrophy and fragility, pigmentation and hair loss.

[0006] However, although type XVII collagen plays an important regulatory role in the self-purification of skin cells, supplementing type XVII collagen can maintain the self-purification process of young cells and effectively slow down the aging process of the skin, type XVII collagen is a large molecular transmembrane protein, and the preparation of heterologous full-length and transdermal absorption are challenges for its application as an anti-aging product. Summary of the invention

[0007] To solve the above problems, the present application provides an oligopeptide that can induce collagen synthesis in vivo or simulate protein regulatory functions. According to the discovery of the present application, in type XVII collagen, the core region that transmits regulatory signals is near the C-terminus of the protein. By analyzing, retaining and disassembling the key domains, regulatory function domain fragments similar to type XVII collagen can be obtained, i.e., the oligopeptides in the present application. The oligopeptide contains the key functional part of type XVII collagen, and has the function of regulating the self-purification and renewal of aging, damaged and infected cells in the skin.

[0008] Specifically, the present invention relates to the following:

[0009] An oligopeptide or a derivative thereof, wherein the oligopeptide has a structural formula as shown in the following formula (1):

[0010] (X1)(R or K or H)G(D or E)#(X2)(1)

[0011] Wherein, X1 represents an amino acid selected from G, A, V, L, I, and P or represents the absence of any amino acid,

[0012] # represents an amino acid selected from A, V, D, E, N, Q,

[0013] X2 represents G, P or the absence of any amino acid.

[0014] Furthermore, in the structural formula of the oligopeptide, X1 is chemically modified by a water-soluble group or an oil-soluble group. Preferably, the water-soluble group or the oil-soluble group is an acetyl group, a tert-butyryl group, a hexanoyl group, a 2-methylhexanoyl group, an octanoyl group, a decanoyl group, a lauroyl group, a myristoyl group, a palmitoyl group, a stearoyl group, an oleoyl group or a linoleoyl group.

[0015] Furthermore, the present application provides an oligopeptide having an amino acid sequence as shown in RGD.

[0016] Furthermore, the present application provides an oligopeptide having an amino acid sequence as shown in PRGEA (Seq ID No. 1).

[0017] Furthermore, the present application provides an oligopeptide having an amino acid sequence as shown in PRGEQG (Seq ID No. 2).

[0018] The present application also provides the use of the oligopeptide or its derivatives in clearing senescent fibroblasts, inhibiting collagen degradation, increasing collagen precursors, upregulating collagen expression, or regulating the expression of cellular inflammatory factors or repair factors.

[0019] The present application also provides the use of the oligopeptide or its derivatives in the preparation of cosmetics or pharmaceutical compositions or health products for anti-aging, anti-inflammatory, whitening, promoting skin repair and regeneration or improving skin allergies.

[0020] The present application also provides a composition for anti-aging, anti-inflammatory, whitening, promoting skin repair and regeneration or improving skin allergies, which comprises one or more of the oligopeptides or their derivatives.

[0021] Furthermore, the composition also includes a cosmetically or pharmaceutically acceptable carrier or excipient or a second active ingredient.

[0022] Furthermore, the second active ingredient in the composition is selected from active ingredients having anti-aging, moisturizing or whitening effects.

[0023] The present application also provides a cosmetic or pharmaceutical composition or health product, which comprises one or more of the oligopeptides or their derivatives.

[0024] Effects of the Invention

[0025] The oligopeptides provided in this application have the following beneficial effects:

[0026] In cell experiments, it can effectively remove senescent fibroblasts, inhibit the expression of collagen degrading enzymes, increase collagen precursors, and act as a signal molecule to upregulate the expression of collagen, achieving the initial anti-aging effect.

[0027] It can regulate the expression of cellular inflammatory factors and / or repair factors to achieve the two-in-one effect of anti-inflammatory and repair;

[0028] The molecular weight is only 500-600Da, and it can be absorbed through the skin. It can promote the self-cleaning and renewal of epidermal cells just by applying it.

[0029] It can even out skin tone, reduce fine lines, increase dermis thickness, and improve skin redness and allergies;

[0030] Can accelerate wound healing and strengthen skin barrier;

[0031] Oligopeptides at working concentrations are mild and low-irritating, with no obvious adverse reactions in human experiments, and are a safe and effective anti-aging ingredient. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] Figure 1 Shown is the expression gel map in Example 1

[0033] Figure 2 Shown are the experimental results of the toxic effects of the oligopeptide samples on HaCaT cells in Example 2.2.

[0034] Figure 3 Shown are the results of the antagonistic test of the oligopeptide samples in Example 2.3 on the cell aging model.

[0035] Figure 4a —4e shows the experimental results of the oligopeptide-induced synthesis effect on various types of collagen in Example 3.

[0036] Figure 5 Shown are the results of the qPCR experiment of anti-inflammatory factors in Example 4.1.

[0037] Figure 6 Shown are the expression results of inflammation-related genes in the RNA-Seq experiment in Example 4.2.

[0038] Figure 7 , Figure 8 , Fig. 9 Shown are the results of the cell scratch experiment in Example 5.

[0039] Fig.10 Shown is the H&E staining image of the cells in Example 6.

[0040] Fig.11Shown is a trend chart of cell tissue activity in Example 6.

[0041] Fig.12 Shown are the results of the in vitro tyrosinase inhibition experiment in Example 7. DETAILED DESCRIPTION

[0042] The present application provides oligopeptides with anti-aging skin care effects, which can promote cell self-purification, help remove aging skin cells, reconstruct skin rhythms, maintain good skin structure, resist early aging, promote skin repair and regeneration, and reduce fine lines. The oligopeptides can be prepared by existing chemical and biological methods and applied to daily chemicals, food and health care fields. The present application is further described in detail below in conjunction with specific embodiments. The embodiments given are to enable a more thorough understanding of the present application and to enable the scope of the present application to be fully conveyed to those skilled in the art, but they cannot be understood as limiting the scope of protection of the present invention.

[0043] It should be noted that certain words are used in the specification and claims to refer to specific components. Those skilled in the art should understand that technicians may use different nouns to refer to the same component. This specification and claims do not use the difference in nouns as a way to distinguish components, but use the functional differences of components as the criterion for distinction. As mentioned throughout the specification and claims, "including" or "comprising" are open-ended terms and should be interpreted as "including but not limited to". The subsequent description of the specification is a preferred embodiment of the present application, but the description is based on the general principles of the specification and is not intended to limit the scope of the present application. The scope of protection of the present application shall be determined by the attached claims.

[0044] The present application provides an oligopeptide, the structural formula of which is shown in the following formula (1):

[0045] (X1)(R or K or H)G(D or E)#(X2)(1)

[0046] Wherein, X1 represents an aliphatic amino acid, such as one of the amino acid sequences G, A, V, L, I, and P,

[0047] # represents an acidic amino acid or its amide form, such as one of the amino acid sequences A, V, D, E, N, Q,

[0048] X2 represents the amino acid sequence G, P, or the absence of an amino acid.

[0049] In the aforementioned oligopeptide sequences, capital letters such as G, A, V, L, I, D, E, N, Q, and P each represent an amino acid or its amino acid residue, and the correspondence between the capital letters and the amino acids is shown in Table 1.

[0050] Table 1 Amino acids and their corresponding capital letters

[0051] Chinese name Alphabet abbreviation Chinese name Alphabet abbreviation Glycine G Serine S Alanine A Threonine T Valine V Cysteine C Leucine L Asparagine N Isoleucine I Glutamine Q Proline P Aspartic acid D Phenylalanine F Glutamate E Tryptophan W Lysine K Methionine M Arginine R Tyrosine Y Histidine H

[0052] In a specific embodiment of the present application, the amino acid sequence of the oligopeptide is RGD.

[0053] In a specific embodiment of the present application, the amino acid sequence of the oligopeptide is PRGEA (Seq ID No. 1).

[0054] In a specific embodiment of the present application, the amino acid sequence of the oligopeptide is PRGEQG (Seq ID No. 2).

[0055] The present application also provides derivatives of the oligopeptide as shown in structural formula (1).

[0056] In an embodiment of the present application, the derivative of the oligopeptide includes a chemically modified product, a polymer or a cosmetic or pharmaceutically acceptable salt of the oligopeptide.

[0057] In some embodiments, the oligopeptide derivative is a compound obtained by chemically modifying X1 in the oligopeptide structural formula (1). In a specific embodiment, X1 can be optionally chemically modified with a water-soluble or oil-soluble group. For example, X1 can be modified with an acetyl group, a tert-butyryl group, a hexanoyl group, a 2-methylhexanoyl group, an octanoyl group, a decanoyl group, a lauroyl group, a myristoyl group, a palmitoyl group, a stearoyl group, an oleoyl group, or a linoleoyl group.

[0058] In other embodiments, the oligopeptide derivative is a cosmetically acceptable salt of the oligopeptide of formula (1), or a cosmetically acceptable salt thereof, or a pharmaceutically acceptable salt thereof. For example, it includes a metal salt of the oligopeptide shown in formula (1), wherein the metal includes, but is not limited to, lithium, sodium, potassium, calcium, magnesium, manganese, copper, zinc or aluminum, etc.; includes a salt formed by the oligopeptide shown in formula (1) and an organic base, wherein the organic base includes, but is not limited to, ethylamine, diethylamine, ethylenediamine, ethanolamine, diethanolamine, arginine, lysine, histidine or piperazine, etc.; includes a salt formed by the oligopeptide shown in formula (1) and an inorganic acid or an organic acid, wherein the organic acid includes, but is not limited to, acetic acid, citric acid, lactic acid, malonic acid, maleic acid, tartaric acid, fumaric acid, benzoic acid, aspartic acid, glutamic acid, succinic acid, oleic acid, trifluoroacetic acid, oxalic acid, pamoate or gluconic acid, etc.; the inorganic acid includes, but is not limited to, hydrochloric acid, sulfuric acid, boric acid or carbonic acid.

[0059] In other embodiments, the derivative of the oligopeptide is a conjugate of the oligopeptide and a carrier, or a mixture comprising the oligopeptide, or a polypeptide comprising the oligopeptide sequence, or a polymer, a cyclic polypeptide, etc. of the oligopeptide.

[0060] The oligopeptides or derivatives thereof provided in the present application can be prepared by chemical and biological methods in the prior art. In some specific embodiments, they can be prepared by chemical solid phase synthesis, liquid phase synthesis or a combination of solid phase and liquid phase, or by sequence-targeted biotechnology methods, cell factory recombinant expression or initial separation of type XVII collagen or protein fragments by hydrolysis.

[0061] In a specific embodiment, the oligopeptide or its derivative can be synthesized by a cell factory. The specific steps include:

[0062] The oligopeptide is fused to assist in the expression of a purified protein tag as a heterologous synthetic target protein, which is cultured and expressed using genetically engineered bacteria. The expressed target protein is preliminarily purified by affinity chromatography, the purified protein is cleaved by protease to remove its tag, and ultrafiltration / nanofiltration is used to separate and purify the oligopeptide.

[0063] In some specific embodiments, the cell factory can be a prokaryotic expression host such as Escherichia coli, a eukaryotic expression host such as yeast cells, or a plant reactor, etc.

[0064] The present application also provides the use of an oligopeptide having a composition as shown in formula (1) or its derivatives in removing senescent fibroblasts, inhibiting collagen degradation, increasing collagen precursors, upregulating collagen expression, regulating the expression of cellular inflammatory factors and repair factors, or promoting microwound repair.

[0065] In a specific embodiment, the oligopeptides or their derivatives provided by the present application can be applied to the fields of daily chemicals, food and health care.

[0066] Specifically, the present application provides the use of an oligopeptide having a composition as shown in formula (1) or a derivative thereof in the preparation of cosmetics for anti-aging, anti-inflammatory, promoting skin repair and regeneration, or improving skin allergies.

[0067] The present application also provides the use of an oligopeptide having a composition as shown in formula (1) or a derivative thereof in the preparation of a pharmaceutical composition for anti-aging, anti-inflammatory, promoting skin repair and regeneration or improving skin allergies.

[0068] In a specific embodiment, the oligopeptide or its derivative can effectively remove aging skin cells, reconstruct skin rhythm, maintain good skin structure, resist early aging, promote skin repair and regeneration, reduce skin fine lines, and improve skin allergies by promoting cell self-purification.

[0069] The present application also provides the use of an oligopeptide having a composition as shown in formula (1) or a derivative thereof in the preparation of a health product for anti-aging, anti-inflammatory, promoting skin repair and regeneration or improving skin allergies.

[0070] The present application also provides a composition for anti-aging, anti-inflammatory, promoting skin repair and regeneration or improving skin allergies, which comprises one or more oligopeptides or their derivatives whose composition is shown in formula (1). Preferably, the sequence of the oligopeptide is RGD, PRGEA (Seq ID No. 1) or PRGEQG (Seq ID No. 2).

[0071] In a specific embodiment, the composition further comprises a cosmetically or pharmaceutically acceptable carrier or excipient or a second active ingredient.

[0072] In some specific embodiments, the second active ingredient is selected from active ingredients having anti-aging, moisturizing or whitening effects.

[0073] In a specific embodiment of the present application, the oligopeptide or its derivative, or a composition comprising the oligopeptide or its derivative, can be delivered in an organism via a delivery system. Specifically, the delivery system can be selected from: liposomes, oleosomes, nonionic surfactant liposome vesicles, ethosomes, millicapsules, microcapsules, nanocapsules, nanostructured lipid carriers, sponges, cyclodextrins, lipid vesicles, micelles, millispheres, microspheres, nanospheres, lipid spheres, microemulsions, nanoemulsions, milliparticles, microparticles or nanoparticles; in some preferred embodiments, it can be selected from liposomes or microemulsions; in other preferred embodiments, it can be selected from water-in-oil microemulsions with an internal structure of reverse micelles.

[0074] In a specific embodiment, the oligopeptide or its derivative can be used to prepare a purified component solution for cosmetics or to prepare a mixed stock solution.

[0075] The present application also provides cosmetics or body care products comprising the oligopeptide or its derivatives. In the embodiments of the present application, there is no limitation on the form of the cosmetics or body care products, which may be facial cleansers, shower gels, lotions, skin care lotions, skin care creams, essences, eye creams, facial masks, aerosols / sprays, gels, freeze-dried powders, soaps, shower gels, shampoos, shaving creams, hand sanitizers, or any other forms conventionally applicable in the field of cosmetics or body care.

[0076] Furthermore, the cosmetic or body care product also includes cosmetically acceptable excipients, such as one or a combination of two or more of moisturizers, oils, emulsifiers, thickeners, preservatives, antioxidants, skin feel regulators, fragrances and solvents.

[0077] In a specific embodiment, the moisturizer can be a commonly used moisturizer in the art, for example, can be selected from: one or more of glycerin, allantoin, hexylene glycol, glyceryl polyether-26, panthenol, propylene glycol, hyaluronic acid and its salts (such as sodium hyaluronate).

[0078] In a specific embodiment, the oil can be a commonly used oil in the art, for example, it can be selected from animal and vegetable oils and fats, higher fatty acids, hydrocarbons, esters, waxes, higher alcohols, shrimp goby oils, silicone substances, sterols and resins, as well as the above substances treated by enzymes or chemicals.

[0079] In a specific embodiment, the emulsifier can be an emulsifier commonly used in the art, for example, it can be selected from: sorbitan olivate, hydrogenated phospholipids, PEG-100 stearate, cetearyl alcohol, glyceryl stearate, alkyl sulfates, dodecylbenzene sulfonates, lactates, sulfosuccinates, monoglyceride sulfonates, phosphates, silicones, and taurates. One or more of the following.

[0080] In a specific embodiment, the thickener can be a commonly used thickener in the art, for example, can be selected from one or more of gum arabic, pectin, agar, gelatin, carbomer, soluble starch, polysaccharide derivatives, carboxymethyl cellulose, propylene glycol alginate, methyl cellulose, sodium starch phosphate, sodium carboxymethyl cellulose, sodium alginate, casein, sodium polyacrylate, polyoxyethylene, polyvinyl pyrrolidone, etc.

[0081] In a specific embodiment, the preservative may be a preservative commonly used in the art, for example, one or more selected from sorbic acid, caprylhydroxamic acid, benzyl alcohol, benzoic acid, glyceryl caprylate, ethylhexylglycerin, and the like.

[0082] In a specific embodiment, the antioxidant can be a commonly used antioxidant in the art, for example, it can be selected from one or more of astaxanthin, vitamin C, vitamin E, glutathione, lipoic acid, uric acid, ectoine, butylated hydroxymethyl (BHT), fullerene, butylated hydroxyanisole (BHA), carotene or panthenol (coenzyme Q).

[0083] In a specific embodiment, the skin feel regulator can be a commonly used skin feel regulator in the art, for example, can be selected from: one or more of sesame oil, sunflower seed oil, dendrobium polysaccharide, etc.

[0084] In a specific embodiment, the fragrance can be a commonly used fragrance in the art, for example, can be selected from: plant fragrances (floral fragrance, fruity fragrance, green fragrance), animal fragrances (musk, ambergris), synthetic fragrances (menthol, camphor), etc.

[0085] The present application also provides a pharmaceutical composition comprising the oligopeptide or its derivative. Specifically, the pharmaceutical composition can be prepared in the following forms: oral preparations, such as tablets (including sugar-coated tablets, film-coated tablets, sublingual tablets, orally disintegrating tablets), capsules (including soft capsules, microcapsules), granules, powders, lozenges, syrups, emulsions, suspensions, films (e.g., films of oral disintegrating properties), etc., parenteral preparations such as injections (e.g., subcutaneous injections, intravenous injections, intramuscular injections, intraperitoneal injections, drops), topical preparations (e.g., skin preparations, ointments), suppositories (e.g., rectal suppositories, vaginal suppositories), pills, nasal drops, respiratory preparations (inhalants), eye drops, etc. In addition, these preparations can be used as controlled release preparations (e.g., sustained release microcapsules), such as immediate release preparations, sustained release preparations, etc. Such preparations can be obtained by the preparation methods conventionally used in the art.

[0086] Furthermore, the pharmaceutical composition also includes a pharmaceutically acceptable carrier, such as: an excipient (e.g., starch, lactose, sucrose, calcium carbonate, calcium phosphate, etc.), a binder (e.g., starch, gum arabic, carboxymethyl cellulose, hydroxypropyl cellulose, crystalline cellulose, alginic acid, gelatin, polyvinyl pyrrolidone, etc.), a lubricant (e.g., magnesium stearate, calcium stearate, talc, etc.), a disintegrant (e.g., carboxymethyl cellulose calcium, talc, etc.), a diluent (e.g., water for injection, saline, etc.), an additive (e.g., a stabilizer, a preservative, a colorant, a flavoring agent, a dissolution aid, an emulsifier, a buffer, an isotonic agent, etc.), and the like.

[0087] Example

[0088] Unless otherwise specified, the materials, reagents, etc. used in this application can be obtained from commercial sources.

[0089] Example 1 Preparation and determination of anti-aging oligopeptides

[0090] The oligopeptides in this application can be synthesized by fusing tags that assist expression and purification. The specific method is:

[0091] According to their amino acid sequences, the nucleotide sequences of oligopeptides RGD, PRGEA (Seq ID No. 1), and PRGEQG (Seq ID No. 2) were connected to the pGEX-4t-1 vector by overlap PCR for expression. The constructed oligopeptide expression vectors were transformed into an Escherichia coli host and the proteins were expressed.

[0092] E. coli containing the expression vector was cultured in LB medium at 37°C and 220 rpm. 600When a certain value is reached, an inducer is added to the culture medium to induce protein expression. The expressed bacteria are collected by centrifugation, broken, and centrifuged. The cell fragments are purified by affinity chromatography, cut, and oligopeptides are obtained and identified. The oligopeptide samples obtained are TYPE1 (RGD), TYPE2 (PRGEA), and TYPE4 (PRGEQG). Figure 1 Shown is an expression gel image.

[0093] Unless otherwise specified, TYPE1, TYPE2, and TYPE4 used in the following examples are the samples obtained in this example.

[0094] Example 2 Anti-aging effect test of oligopeptides

[0095] 2.1 Establishment of H2O2-induced HaCaT cell senescence model

[0096] Experimental principle: Oxidative stress (OS) is a pathological state in which excessive production of reactive oxygen species (ROS) in the body leads to an imbalance in the oxidation and antioxidant systems, causing oxidative damage to cells or tissues. ROS causes oxidative damage to DNA, proteins, lipids, etc., destroys the integrity of cell structure and function, and causes cell apoptosis or necrosis. As one of the reactive oxygen species, H2O2 is not only extremely easy to penetrate the cell membrane and react with intracellular iron ions to generate highly active free radicals, but also easy to obtain and stable in nature. It has become an important tool for studying oxidative damage to various types of cells at home and abroad. The HaCaT cell aging model was established by inducing HaCaT oxidative stress with H2O2, which was subsequently used to test and evaluate the anti-aging effects of oligopeptide samples.

[0097] Experimental method: HaCaT cells in the logarithmic growth phase were taken and cultured at 5-8×10 3 Each well was inoculated with 100 μL of H2O2 and incubated for 24 hours in a 5% CO2 incubator. The culture medium was removed after 24 hours, and the cells were washed twice with PBS (pH=7.4 buffer). Serum-free medium (DMEM) was added and cultured overnight. The culture medium was discarded after 24 hours, and the cells were washed twice with PBS. Different concentrations of H2O2 were added to each well and cultured for 24 hours. The culture medium was discarded, and the cells were washed twice with PBS. The MTT method (100 μL MTT (1.0 g·L -1 ) solution, incubate at 37℃, 5% CO2 for 4h, discard the solution, add 100μL DMSO, and place at 37℃ for 30min) Measure the absorbance of each well with a microplate reader at a wavelength of 490nm, calculate the cell survival rate, and observe the cell state. Select the H2O2 concentration with a cell survival rate of about 50% to establish a cell senescence model in preparation for subsequent experiments.

[0098] The cell survival rate calculation formula is:

[0099] V(%)=(OD 样品 -OD 空白对照 ) / (OD 细胞对照 -OD 空白对照 )×100%

[0100] Where: V (%) - cell viability, %; OD 样品 ——Absorbance of the reaction system containing the sample to be tested; OD 空白对照 ——Absorbance of an empty plate without any substance; OD 细胞对照 ——The absorbance of the reaction system without the sample to be tested.

[0101] Experimental results: An aging model was established with a H2O2 concentration of (50μM / L) as the model group in experiment 2.3.

[0102] 2.2 Experiment on toxicity of oligopeptide samples on HaCaT cells

[0103] Experimental principle: Succinate dehydrogenase in the mitochondria of living cells can reduce exogenous MTT to water-insoluble blue-purple crystalline formazan and deposit it in the cells, while dead cells do not have this function. Dimethyl sulfoxide (DMSO) can dissolve the formazan in the cells, and its light absorption value is measured at a wavelength of 490nm using an enzyme-linked immunosorbent assay, which can indirectly reflect the number of living cells. By detecting the survival rate of cells under different concentration conditions, the optimal sample concentration for subsequent experiments can be screened.

[0104] Experimental method: HaCaT cells in the logarithmic growth phase were taken and (3-5)×10 3 Each well was inoculated with 100 μL of DMEM medium and cultured for 24 h. The cells were washed twice with PBS and the MTT method (100 μL MTT (1.0 g·L) was used to detect the presence of 100 μL of TYPE1, TYPE2 and TYPE4 oligopeptide samples) was used. -1 ) solution, incubate at 37°C, 5% CO2 for 4 hours, discard the solution, add 100 μL DMSO, and place at 37°C for 30 minutes) Measure the absorbance of each well with a microplate reader at a wavelength of 490 nm to calculate the cell survival rate. The sample concentration with a cell survival rate ≥ 100% was selected as the subsequent test concentration.

[0105] The cell survival rate calculation formula is:

[0106] V(%)=(OD 样品 -OD 空白对照 ) / (OD细胞对照 -OD 空白对照 )×100%

[0107] Where: V (%) - cell viability, %; OD 样品 ——Absorbance of the reaction system containing the sample to be tested; OD 空白对照 ——Absorbance of an empty plate without any substance; OD 细胞对照 ——The absorbance of the reaction system without the sample to be tested.

[0108] Experimental results: Figure 2 As shown, the cell viability of each oligopeptide sample at three concentrations (10 / 50 / 100 ppm) was ≥100%, which can be used as the action concentration for subsequent experiments.

[0109] 2.3 Anti-aging test of oligopeptide samples on cell aging model

[0110] A sample group (5 replicates), a model group and a control group were set up for comparative experiments to evaluate the protective effect of each oligopeptide sample on the HaCaT cell aging model.

[0111] Experimental method: Take cells in logarithmic growth phase and calculate (8-10)×10 3 Each well was inoculated with 100 μL of reagent 1 and incubated for 24 h in a 5% CO2 incubator. The cells were cultured overnight at 37°C and 5% CO2. The supernatant was discarded and the cells were washed twice with PBS. Each group was added with 100 μL of reagent 2 and incubated for 24 h. The supernatant was discarded and the cells were washed twice with PBS. The cells were then cultured using the MTT method (100 μL of MTT (1.0 g·L -1 ) solution, placed in a 37°C, 5% CO2 environment for 4 hours, discarded, added 100 μL DMSO, placed at 37°C for 30 minutes) and measured the absorbance of each well with an ELISA reader at a wavelength of 490 nm, calculated the cell survival rate, and observed the cell status. The composition of reagent 1 and reagent 2 in each group is shown in Table 2:

[0112] Table 2 Composition of reagents in each group in the confrontation test

[0113]

[0114] Experimental results: Figure 3 As shown, the cell survival rates of the oligopeptide samples at concentrations of 50 and 100 ppm were greater than those of the model group, indicating that they all had effective protective effects on the aging model. Among them, the cell survival rate of TYPE4 oligopeptide at 50 ppm was greater than that of the control sample acetyl hexapeptide-8, and the effect was the best.

[0115] Example 3 Experiment on the effect of oligopeptides in inducing collagen synthesis

[0116] Experimental method: HaCaT cells with good growth status were cultured at a rate of about 5×10 5 The cells were inoculated in a culture plate at a density of 1×10 cells / mL and cultured overnight in a DMEM+10% serum+1% double antibody medium at 37°C and 5% CO2. The culture medium was replaced and different concentrations of TYPE2 and TYPE4 samples (10ppm, 50ppm) were added. Five parallels were set for each concentration. DMEM culture medium without samples was added to the blank control group. HaCaT cells were cultured in a six-well plate until 1×10 cells / mL were cultured. 6 After digestion with trypsin, the cells were washed with PBS and 500 μL of Trizol was added and pipetted several times until the cells were lysed. The total RNA was extracted from the lysed cells using chloroform and ethanol. The extracted total RNA was reverse transcribed and qPCR was performed using a reverse transcription kit (AT311-03) and a real-time fluorescence quantitative PCR kit to measure the transcription level of the protein. The experimental results are shown in Figure 4a —4e (in the figure, 0 represents the control group, 2-10 represents the TYPE2 sample group with a concentration of 10 ppm, 2-50 represents the TYPE2 sample group with a concentration of 50 ppm, 4-10 represents the TYPE4 sample group with a concentration of 10 ppm, and 4-50 represents the TYPE4 sample group with a concentration of 50 ppm).

[0117] Analysis of experimental results: Compared with the blank control group, the transcription levels of both chains of type I collagen were significantly increased in the experimental groups with the addition of 10ppm and 50ppm oligopeptide samples.

[0118] In the TYPE4 sample group, the transcription levels of COL1A1 in the 10ppm and 50ppm samples were 1.5 times and 3.2 times that of the control group, and the transcription levels of COL1A2 were 2.3 times and 1.6 times that of the control group. At the same time, the addition of oligopeptide samples can effectively promote the synthesis of type III and type XVII collagen. Among them, the transcription levels of COL3A1 in the 10ppm and 50ppm samples were 3.7 times and 5.4 times that of the control group, and the transcription levels of COL17A1 were 0.84 times and 1.6 times that of the control group.

[0119] At the same time, in the 10ppm and 50ppm experimental groups, the collagen degrading enzyme decreased to 0.73 and 0.79 times of the control group.

[0120] In the TYPE2 sample group, the transcription levels of COL1A1 in the 10ppm and 50ppm samples were 1.1 and 1.3 times of the control group, the transcription levels of COL3A1 were 2.4 and 1.3 times of the control group, and the transcription levels of COL17A1 were 1.5 and 0.97 times of the control group. At the same time, the collagen degrading enzyme in the 10ppm and 50ppm experimental groups decreased to 0.53 and 0.75 times of the control group.

[0121] From the above cell experiments, it can be seen that the oligopeptides in this application can effectively promote the synthesis of type I, type III and type XVII collagen, while reducing the expression of collagenase. Therefore, it can effectively increase the supporting structure collagen type I and type III collagen, and the skin regeneration signal type XVII collagen, thereby achieving an anti-aging effect.

[0122] Example 4 Anti-inflammatory effect experiment of oligopeptides

[0123] 4.1 Anti-inflammatory factor qPCR experiment

[0124] Experimental method: HaCaT human immortalized keratinocytes in good growth condition were cultured at 5×10 5 Cells / well were inoculated in a 6-well plate and cultured in a 37°C 5% CO2 incubator; the culture medium was discarded, and TYPE2 oligopeptide samples of different concentrations (10 / 50 ppm) were added and cultured for 48 hours; the culture medium was discarded, and the cell pellets were collected after washing with PBS twice; RNA was extracted, reverse transcribed into cDNA, and then qPCR was performed.

[0125] like Figure 5 As shown, the qPCR experimental results showed that the expression level of inflammatory factor IL-6 was significantly decreased.

[0126] 4.2 Anti-inflammatory factor RNA-Seq experiment

[0127] Experimental method: HFF-1 human fibroblasts in good growth condition were cultured at 1.5×10 6 Cells were inoculated into 6-well plates and cultured in a 37°C 5% CO2 incubator. The culture medium was discarded, and samples of different concentrations were added and cultured for 48 hours. The culture medium was discarded, and the cells were washed twice with PBS, and then the cell clusters were collected for subsequent transcriptomic analysis.

[0128] like Figure 6 As shown, the results of RNA-Seq experiments showed that the expression levels of inflammation-related genes were all reduced.

[0129] Example 5 Experiment on the wound repair effect of oligopeptides

[0130] The wound healing effect of oligopeptides was tested by cell scratch test. The cell scratch (wound healing) method is a simple method to measure cell migration and repair ability, similar to the in vitro wound healing model. On the monolayer adherent cells cultured in vitro culture dishes or plates, a line is drawn in the central area of ​​cell growth with a micropipette tip or other hard objects, the cells in the central part are removed, and then the cells are cultured to the time set in the experiment. The cell culture plate is taken out and the growth and migration ability of the surrounding cells is observed. The experiment sets a normal control group and an experimental group. The experimental group is a group with some treatment factors or drugs, exogenous genes, etc. The migration and repair ability of each group of cells can be judged by the difference in the repair ability of the scratch area between the cells in different groups.

[0131] Experimental method: Immortalized human keratinocytes in good growth condition were cultured at 1×10 6 The cells were seeded in a 6-well plate and cultured in an incubator at 37°C and 5% CO2. On the second day (when the cells were observed to be fully grown), a 10μL pipette tip was used to measure the ruler and draw two vertical cross lines (+ shape), and the cells were washed 3 times with PBS to remove the scratched cells. Subsequently, 2mL of TYPE1 oligopeptide samples of different concentrations were added to each well as the experimental group, and serum-free DMEM culture medium was used as the control group. After the cells in the experimental and control groups were loaded, they were continued to be cultured in an incubator at 37°C and 5% CO2, and observed and photographed under an inverted microscope at 0, 6, 12, and 24 hours after loading. ImageJ software was used for data analysis to calculate the scratch area value and cell migration rate. The results are shown in Table 3-1 and Figure 7 The same experimental method was used to test TYPE2 and TYPE4 oligopeptide samples of different concentrations to obtain the cell migration rate values ​​of each sample group. The results are shown in Table 3-2 and Figure 8 , 9 shown.

[0132] Table 3-1 Repair effect of TYPE1 oligopeptide on HaCaT cell scratches

[0133]

[0134] Table 3-2 Repair effect of TYPE2 and 4 oligopeptides on HaCaT cell scratches

[0135]

[0136] Analysis of experimental results: The photographic results were analyzed using Image J software to calculate the scratch area value and cell migration rate in each group to evaluate the wound repair ability of each experimental group. The experimental results showed that in the experimental group with the addition of oligopeptides, the scratch area recovery and cell migration rate were faster. Therefore, it can be proved that the oligopeptide has an effective effect in promoting microwound healing.

[0137] Example 6 Full-thickness skin model experiment with UVR irradiation

[0138] The sample group, positive control group, negative control group and blank control group were set up to conduct full-thickness skin model under UVR irradiation. Test. The skin model and the corresponding culture medium were provided by Microspectro Testing Company. The test indicators were skin tissue morphology and tissue vitality to evaluate the resistance and repair ability of oligopeptide samples to UVA and UVB damage. The experimental conditions of each group are shown in Table 4.

[0139] Table 4 UVR irradiation experiment settings for each group

[0140]

[0141]

[0142] Experimental method: The model was transferred to a 6-well plate (2.0 mL of the model culture medium of the corresponding group was added in advance), and the test group number was marked on the 6 wells. Incubate in a CO2 incubator (37°C, 5% CO2) for 24 h. UVR irradiation and drug administration were performed according to the treatment conditions of each group in Table 4: the UVR irradiation dose was UVA (30 J / cm 2 ) and UVB (50mJ / cm 2 ), irradiated continuously for 4 days, replaced with fresh culture medium after each irradiation, and drug treatment was performed. The samples to be tested were administered by surface drug administration. After 4 days of continuous irradiation, the whole layer of skin tissue was collected for testing.

[0143] Experimental results and analysis:

[0144] (1) Tissue morphology detection: The model was fixed with 4% paraformaldehyde. After fixation for 24 hours, the model was removed by circumcision and H&E staining was performed. The model was observed under a microscope and pictures were collected. Fig.10 shown.

[0145] Compared with the NC group, the damaged condition of the model in the PC (VC+VE) group was significantly improved, and the number of damaged cells was significantly reduced, indicating that this positive control test was effective.

[0146] Compared with the NC group, the damage of the model in the sample TYPE4 group was improved to a certain extent, and the number of damaged cells was significantly reduced.

[0147] Therefore, based on the 3D skin model Compared with the NC group, at 100% administration concentration, the damage of the model in the sample TYPE4 group was improved to a certain extent, and the damaged cells were significantly reduced, indicating that the sample TYPE4 has a certain effect in resisting UVA+UVB damage.

[0148] (2) Tissue Viability Detection: After the model is cleaned, prepare a 6-well plate according to the number of models and mark it accordingly. Add 0.3 mL of MTT working solution (1.0 g L -1 ). Place the 24-well plate containing the model in an incubator (37±1℃, 5±1% CO2) and incubate for 3h±5min. Wash the outer surface of the model with PBS, wipe it dry with a sterile cotton swab, transfer it to a new 6-well plate, and mark it. Add 2mL of isopropanol to the inner surface of the model, seal the 6-well plate with a sealing film, and shake it on a well plate shaker for 2h. Pierce the model with the tip of a pipette so that the isopropanol extract flows out of the model into the 6-well plate. Discard the pierced model and blow the isopropanol extract in each well 3 times to mix it thoroughly. After mixing, take 2 portions of 200μL of isopropanol extract from each well, add them to the corresponding wells of the 96-well plate, and mark them. Read the absorbance (OD) value at a wavelength of 570nm using an enzyme reader. Calculate the relative tissue viability of each group according to the following formula:

[0149]

[0150] The specific test results are shown in Table 5, and the trend chart of tissue vitality results is shown in Fig.11 shown.

[0151] Table 5 Tissue vitality results

[0152] Group Average tissue vitality (%) SD P-value BC 100.00 11.73 / NC 75.52 12.01 0.007## PC(VC+VE) 97.68 11.41 0.013* 001TYPE4 111.63 10.66 0.005**

[0153] Note: When the t-test method was used for statistical analysis, the significance of the NC group compared with the BC group was indicated by #, P-value < 0.05 was indicated by #, and P-value < 0.01 was indicated by ##; the significance of the sample group compared with the NC group was indicated by *, P-value < 0.05 was indicated by *, and P-value < 0.01 was indicated by **.

[0154] Compared with the blank control group (BC), the tissue activity of the NC group decreased significantly (p<0.01), indicating that the stimulation conditions of this experiment were effective.

[0155] Compared with the NC group, the tissue vitality of the PC group was significantly improved (p<0.01), indicating that the positive control test was effective.

[0156] Compared with the NC group, the tissue viability of the sample TYPE4 group was significantly improved and higher than that of the PC group.

[0157] Example 7 In vitro tyrosinase inhibition effect experiment

[0158] Set up a sample group (different concentrations of oligopeptide sample TYPE4), a control group and a positive control group, prepare each group of reaction systems according to the reagent addition amount in Table 6, mix well, set up 3 replicate wells for each concentration and a background control well, and determine the tyrosinase inhibition rate of each group.

[0159] Table 6 Composition of reaction systems in each group of in vitro tyrosinase inhibition experiment

[0160]

[0161]

[0162] Among them, L-dopa is a substrate and kojic acid is a tyrosinase inhibitor. The sample volume unit of each reagent in Table 6 is μL. The inhibition rate calculation formula is:

[0163] The experimental results are shown in Table 7 and Fig.12 shown.

[0164] Table 7 Analysis of in vitro tyrosinase inhibition test results (Mean±SD)

[0165]

[0166] The results showed that the inhibition rate of oligopeptide sample TYPE4 on tyrosinase at a concentration of 6.25%-100% was statistically different from that of the control group (p<0.05), and the sample had the effect of inhibiting tyrosinase activity.

[0167] Example 8 Human efficacy evaluation experiment

[0168] Eight healthy Chinese female subjects with obvious visible wrinkles at the corners of the eyes, poor skin elasticity, spots and redness on the face were selected, with an age range of 33-53 years old. The before-after control method was used to measure the elasticity and firmness of the subjects' skin before using the test sample, and after using the test sample for 14 days and 28 days respectively, and take full-face photos and local images. The effect of use was judged by calculating the change rate before and after use of the sample. The test sample was TYPE4 oligopeptide. The experiment was commissioned to Shanghai Microspectrum Chemical Technology Service Co., Ltd.

[0169] Experimental method: On the 0th day, the subjects cleaned their faces with a cleansing product and wiped their skin with dry facial tissue. After sitting in a laboratory at a temperature of 21±1℃ and a humidity of 50±10% for 20 minutes, the laboratory technicians operated the visia, Cutometer, and UC22 instruments to measure the relevant basic value indicators of the subjects' test parts. Then the subjects used the product according to the instructions (once in the morning and once in the evening, take an appropriate amount of sample and apply it evenly on the face after each face cleaning, and massage until absorbed). On the 14th and 28th days of use, the subjects were revisited and the relevant indicators were measured (conditions and methods are the same as above), and the subjects completed the self-assessment questionnaire.

[0170] Statistical methods:

[0171] The rate of change is the rate of change relative to before use, and the calculation formula is as follows;

[0172] Δ (difference) on the 14th day of using the product = D14 - D0

[0173] Δ (difference) on the 28th day of using the product = D28-D0

[0174]

[0175] In the above formula, D0 is the basic value of skin parameters before using the product on the affected area; D14 is the value of skin parameters on the 14th day after using the product on the affected area; D28 is the value of skin parameters on the 28th day after using the product on the affected area; N is the number of subjects.

[0176] 8.1 Skin elasticity improvement test

[0177] The skin elasticity tester Cutometer dual MPA580 was used to test the cheek area of ​​the subjects on days 0, 14, and 28, and the elasticity of the skin was determined by analyzing the data of the instrument. The test results of skin elasticity R2 and skin firmness F4 are shown in Table 8.

[0178] Table 8 Skin elasticity and firmness test results

[0179]

[0180] Note: The data in the table are mean ± standard error

[0181] The above experimental results show that the skin elasticity R2 (the higher the value, the more elastic the skin) of the sample application area of ​​the test subjects increased compared with the baseline value at 14 days and 28 days of use, with an increase rate of 5.55% and 20.43% respectively; the skin firmness F4 (the lower the value, the tighter the skin) decreased compared with the baseline value at 14 days and 28 days of use, with a decrease rate of 6.04% and 18.34% respectively. Therefore, it can be proved that the oligopeptide sample can effectively restore skin elasticity and firmness, and has anti-aging effects.

[0182] 8.2 Facial wrinkle improvement test

[0183] The facial image analyzer VISIA was used to take full-face photos of the subjects on days 0, 14, and 28, and the pores, wrinkles, and skin texture of the epidermis were measured and analyzed in real time. The area ratio of wrinkles at the corners of the eyes and under the eyes was detected and analyzed. The test results are shown in Table 9.

[0184] Table 9 Wrinkle area ratio test results

[0185]

[0186] Note: The data in the table are mean ± standard error

[0187] The above experimental results show that the area of ​​wrinkles at the corners of the eyes and under the eyes of the subjects who used the samples decreased compared with the baseline values ​​after 14 and 28 days of use. The area of ​​wrinkles at the corners of the eyes decreased by 16.46% and 28.86% respectively, and the area of ​​wrinkles under the eyes decreased by 23.83% and 49.6% respectively.

[0188] 8.3 Skin pigmentation improvement test

[0189] The face image analyzer VISIA was used to take full-face photos of the subjects on days 0, 14, and 28 to analyze the location of facial spots and the red area on the cheeks, and to measure the changes in the area of ​​ultraviolet spots, brown spots, and red areas. The test results are shown in Table 10.

[0190] Table 10 Detection results of facial spot area ratio

[0191]

[0192] Note: The data in the table are mean ± standard error

[0193] The above experimental results show that the area proportions of brown spots, ultraviolet spots and red zones in the sample use areas of the subjects decreased compared with the baseline values ​​after 14 and 28 days of use. The brown spot area decreased by 6.82% and 14.75% respectively, the ultraviolet spot area decreased by 4.30% and 6.76% respectively, and the red zone area decreased by 6.58% and 15.08% respectively.

[0194] 8.4 Collagen density detection

[0195] The skin ultrasound diagnostic instrument Ultrascan UC22 was used to detect the cheeks of the subjects on days 0, 14, and 28 to obtain skin ultrasound images with high resolution and good contrast, and to analyze the changes in collagen density. The test results are shown in Table 11.

[0196] Table 11 Collagen density test results

[0197]

[0198] Note: The data in the table are mean ± standard error

[0199] The above experimental results show that the collagen density in the area where the samples were used by the subjects increased compared with the baseline value at 14 and 28 days of use. The increase rate was 10.22% at 14 days and 17.70% at 28 days.

[0200] 8.5 Dermis thickness test

[0201] The skin ultrasound diagnostic instrument Ultrascan UC22 was used to detect the cheeks of the subjects on days 0, 14, and 28 to obtain skin ultrasound images with high resolution and good contrast, measure the thickness of the dermis, and analyze its changes. The test results are shown in Table 12.

[0202] Table 12 Dermis thickness test results

[0203]

[0204] Note: The data in the table are mean ± standard error

[0205] The above experimental results show that the thickness of the dermis layer in the area where the samples were used by the subjects increased compared with the baseline value at 14 and 28 days of use. The increase rate was 6.61% at 14 days and 14.88% at 28 days.

[0206] 8.6 Subject Self-Assessment

[0207] The subjects evaluated the samples after 28 days of use. The evaluation indicators and results are shown in Table 13. The scoring criteria are: 1 for "strongly disagree", 2 for "disagree", 3 for "average", 4 for "agree", and 5 for "strongly agree". The "degree of agreement" in Table 7 is the percentage of people who scored ≥ 4 points.

[0208] Table 13 Evaluation of the test subjects on the samples used

[0209] Evaluation indicators Degree of agreement Feel that the product can improve skin firmness after use 75％ I feel that the product can enhance skin elasticity after use 87.5％ I feel that the product can moisturize the skin 87.5％ Feel that the product can improve skin smoothness after use 100％ Feel that the product can reduce fine lines on the skin after use 75％ I feel that the product can achieve anti-wrinkle effects after use 87.5％ Feel that the product can improve skin radiance after use 87.5％ I feel that the product can achieve a firming effect after use 87.5％ This product is gentle and non-irritating 100％ Overall satisfied with this product 87.5％

[0210] Experimental results: After using the test sample for 28 consecutive days, 100.00% of the subjects felt that the product could improve skin firmness after use; 100.00% of the subjects felt that the product could enhance skin elasticity after use. 100.00% of the subjects felt that the product could moisturize the skin. 100.00% of the subjects felt that the product could improve skin smoothness after use. 87.50% of the subjects felt that the product could reduce skin fine lines after use. 87.50% of the subjects felt that the product could achieve anti-wrinkle effect after use. 100.00% of the subjects felt that the product could improve skin radiance after use. 100.00% of the subjects felt that the product could achieve a firming effect after use. 100.00% of the subjects felt that this product was gentle and non-irritating. 100.00% of the subjects were generally satisfied with this product.

[0211] The above is only a preferred embodiment of the present application and does not limit the present application in any form. Any technician familiar with the profession may use the above disclosed technical content to change or modify it into an equivalent embodiment with equivalent changes. However, any simple modification, equivalent change and modification made to the above embodiment based on the technical essence of the present application without departing from the content of the technical solution of the present application still falls within the protection scope of the technical solution of the present application.

Claims

1. An oligopeptide, wherein the amino acid sequence of the oligopeptide is PRGEA (Seq ID No. 1) or RGD.

2. Use of the oligopeptide according to claim 1 in the preparation of cosmetics for anti-aging, whitening, promoting skin repair and regeneration or improving skin allergies.

3. A composition for anti-aging, whitening, promoting skin repair and regeneration or improving skin allergies, comprising the oligopeptide according to claim 1.

4. The composition according to claim 3, characterized in that The composition also includes a cosmetically acceptable carrier or excipient or a second active ingredient.

5. The composition according to claim 4, characterized in that The second active ingredient is selected from active ingredients having anti-aging, moisturizing or whitening effects.

6. A cosmetic or pharmaceutical composition comprising the oligopeptide according to claim 1.

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