Peptides having skin whitening activity and uses thereof

Abstract

The peptide of the present invention has a skin whitening activity by inhibiting the incorporation of melanosomes into keratinocytes and promoting the decomposition of melanosomes. The peptide of the present invention can be used as an active ingredient of a skin whitening cosmetic material, or as an active ingredient of a medicament for treating or preventing hyperpigmentation disorders due to excessive melanosome deposition.

Description

Technical Field

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims the priority of Korean Patent Application No. 10-2022-0155231 filed in the Korean Intellectual Property Office on November 18, 2022, the disclosure of which is incorporated herein by reference in its entirety. Technical Field

[0003] The present invention relates to a peptide having skin whitening activity and use thereof. Background Art

[0004] Skin melanin is present in the skin, hair follicles, eyes, etc., and performs an important function of protecting the skin from ultraviolet rays. However, excessive melanin formation leads to hyperpigmentation diseases such as chloasma, freckles, age spots, etc., causing mental stress and reduced quality of life. Melanin is mainly synthesized in melanocytes located between the basal layers of the epidermis or below the basal layers and in the hair follicles. It is synthesized in melanosomes, which are organelles within melanocytes. These melanosomes move to adjacent keratinocytes through dendrites, and as keratinocytes enter the outer layer of skin, skin color appears. Although the mechanism of how melanosomes move from melanocytes to keratinocytes has not yet been elucidated, it is known that melanosomes are released from there to the outside of the cell after moving to the dendrites of melanocytes, and participate in membrane fusion between melanosomes and keratinocytes and participate in phagocytosis.

[0005] Regarding the synthesis process of melanin, it is ultimately synthesized from L-tyrosine via DOPA, dopaquinone, DOPA pigment, and 5,6-dihydroxyindole (DHI). Traditionally, inhibition of melanin synthesis has focused on factors that inhibit the activity of tyrosinase, an enzyme that catalyzes the rate-controlling step of the melanin synthesis pathway.

[0006] PCT International Publication WO 2020 / 153819 discloses a polypeptide having an activity of inhibiting tyrosinase activity and its use for skin whitening, wherein tyrosinase is an important enzyme in the melanin synthesis pathway. In addition, Korean Patent No. 10-1869783 discloses a peptide having an activity of inhibiting melanin production and tyrosinase activity and its use for skin whitening.

[0007] To date, the development of skin whitening agents has focused on inhibiting the activity of tyrosinase, an important enzyme in melanin synthesis. However, in order to achieve a synergistic effect of skin whitening through various points of action, it is necessary to develop skin whitening agents with different mechanisms or points of action.

[0008] [Prior art literature]

[0009] [Patent Document]

[0010] Patent document 1. WO 2020 / 153819

[0011] Patent Document 2. Korean Patent No. 10-1869783 Summary of the invention

[0012] Technical issues

[0013] The present inventors have conducted research work to develop peptides with improved activity and skin whitening activity. As a result, they have experimentally demonstrated that the novel peptides they have developed have excellent effects of inhibiting melanosome incorporation into keratinocytes and excellent activity of degrading melanosomes, thereby completing the present invention.

[0014] Therefore, an object of the present invention is to provide novel peptides having skin whitening activity.

[0015] Another object of the present invention is to provide a composition for skin whitening, which comprises the peptide having the above-mentioned activity as an active ingredient.

[0016] Another object of the present invention is to provide a pharmaceutical composition for preventing or treating hyperpigmentation diseases, which comprises the peptide having the above-mentioned activity as an active ingredient.

[0017] Another object of the present invention is to provide a cosmetic composition for skin whitening, which comprises the peptide having the above-mentioned activity as an active ingredient.

[0018] Technical Solution

[0019] In order to achieve the above objectives,

[0020] One aspect of the present invention provides a peptide comprising the amino acid sequence of SEQ ID NO:1.

[0021] Another object of the present invention is to provide a composition for skin whitening, which comprises the peptide as an active ingredient.

[0022] Another object of the present invention is to provide a pharmaceutical composition for preventing or treating hyperpigmentation diseases, which comprises the peptide as an active ingredient.

[0023] Another object of the present invention is to provide a cosmetic composition for skin whitening, which comprises the peptide as an active ingredient.

[0024] The present invention will be described in detail below.

[0025] 1. Peptides and their activities

[0026] According to one aspect of the present invention, there is provided a peptide comprising the amino acid sequence disclosed in SEQ ID NO: 1.

[0027] As used herein, the term "peptide" refers to a linear molecule formed by linking amino acid residues together through peptide bonds.

[0028] The peptide of the present invention comprising the amino acid sequence of SEQ ID NO: 1 can be used without modification, but variants or fragments thereof having different amino acid sequences by deletion, insertion, replacement of amino acid residues or a combination thereof may be used within the range not affecting the original activity of the peptide (e.g., skin whitening activity).

[0029] The peptide of the present invention may be modified by phosphorylation, sulfation, acrylation, glycosylation, methylation, farnesylation, etc., within the range that its activity is not changed.

[0030] The peptides of the present invention include peptides containing an amino acid sequence substantially identical to a peptide comprising an amino acid sequence of SEQ ID NO: 1 and variants or active fragments thereof. Substantially identical amino acid sequences refer to amino acid sequences having at least 75%, such as at least 80%, at least 85%, at least 90%, at least 95% and at least 97% sequence identity with the amino acid sequence of SEQ ID NO: 1, respectively. In addition, the peptide may also include a targeting sequence, a tag, a labeled residue, and an amino acid sequence prepared for the specific purpose of increasing the half-life or stability of the peptide.

[0031] The peptides of the present invention may have N-terminal and / or C-terminal modifications that induce the selection of a portion of the amino acid sequence and increase its activity. Through these N-terminal and / or C-terminal modifications, the stability of the peptides of the present invention can be significantly improved, for example, the half-life of the peptide can be increased when administered in vivo. The term "stability" refers to not only in vivo stability, which protects the peptides of the present invention from attack by protein-cleaving enzymes in vivo, but also storage stability (e.g., storage stability at room temperature).

[0032] The N-terminal modification may be the binding of a protective group to the N-terminus of the peptide, wherein the protective group is selected from the group consisting of an acetyl group, a fluoreonylmethoxycarbonyl group, a formyl group, a palmitoyl group, a myristyl group, a stearoyl group, and polyethylene glycol (PEG). The C-terminal modification may be a hydroxyl group (-OH), an amino group (-NH 2 ), azido (-NHNH 2 ) and the like are combined with the C-terminus of the peptide, but are not limited thereto.

[0033] The peptides of the present invention can be prepared by various methods widely known in the art to which the present invention belongs. For example, the peptides of the present invention can be prepared using chemical synthesis methods known in the art, particularly solid phase synthesis techniques (Merrifield, J. Amer. Chem. Soc. 85: 2149-54 (1963); Stewart et al., Solid Phase Peptide Synthesis, 2nd edition Pierce Chem. Co.: Rockford, 111 (1984)) or liquid phase synthesis techniques (U.S. Patent No. 5,516,891).

[0034] The peptide of the present invention has skin whitening activity.

[0035] In one embodiment, the peptide of the present invention has an activity of inhibiting the incorporation of melanosomes in keratinocytes or an activity of promoting the degradation of melanosomes.

[0036] Melanosomes produced in melanocytes move from melanocytes to keratinocytes. In particular, the movement of melanosomes to keratinocytes includes a process of releasing melanosomes from melanocytes and a process of incorporating melanosomes into keratinocytes. The incorporation process of melanosomes can be expressed differently as the uptake of melanosomes by keratinocytes or the phagocytosis of melanosomes by keratinocytes.

[0037] Protease-activated receptor 2 (PAR2), Toll-like receptor 3 (TLR3), and keratinocyte growth factor receptor (KGFR) are involved in the uptake of melanosomes released from melanocytes by keratinocyte phagocytosis.

[0038] In one embodiment, the peptide of the present invention has an activity of inhibiting the expression of one or more genes selected from the group consisting of a protease-activated receptor 2 (PAR2) gene, a Toll-like receptor 3 (TLR3) gene, and a keratinocyte growth factor receptor (KGFR) gene in keratinocytes.

[0039] PAR2 is a receptor protein present in the cell membrane and is known to regulate inflammatory responses, obesity, metabolism and cancer, and to act as a receptor for proteases during infection. The PAR2 protein is present in the membrane of keratinocytes and is involved in the uptake (incorporation) of melanosomes by keratinocytes. It is known that inhibition of PAR2 protein activity leads to inhibition of keratinocyte uptake of melanosomes (Exp Cell Res. 2000 Jan 10; 254 (1): 25-32; Pigment Cell Res. 2001 Aug; 14 (4): 236-42; Exp Dermatol. 2003; 12 Suppl 2: 5-12.).

[0040] TLR3 protein is known to be a receptor protein that recognizes pathogens and plays an important role in activating innate immunity. It is known that stimulation of TRR3 protein in keratinocytes induces improved absorption of melanosomes (J Dermatol Sci. 2019Dec;96(3):168-177), and it is also known that in melanocytes, TRR3 protein promotes the migration of melanosomes into keratinocytes (Int. J. Mol. Sci. 2020, 21, 9769).

[0041] It is known that KGFR, which is a tyrosine kinase receptor, is expressed in many types of epithelial cells, activated by four known ligands (i.e., FGF-1, FGF-3, FGF-7, and FGF-10), and is involved in the proliferation and differentiation of epithelial cells and wound recovery. In addition, KGFR is also known to promote the migration of melanosomes into keratinocytes (J Invest Dermatol. 2005 Dec; 125(6): 1190-9., Journal of Investigative Dermatology Vol. 128, No. 3, March 2008, p. 558-567).

[0042] In one embodiment, the peptide of the present invention has an activity of upregulating the amount of Beclin-1 protein or LC3-I / II protein in keratinocytes.

[0043] Autophagy is a natural process that removes unnecessary, dysfunctional or ineffective materials (such as denatured proteins) from cells. For example, organelles that have degraded and become dysfunctional at the end of their lifespan are removed by autophagy. Proteins or organelles that undergo autophagic removal are sequestered in double-membrane vesicles called autophagosomes within the cell, and these vesicles are again combined with lysosomes to form autophagolysosomes, in which they are degraded by lysosomal enzymes.

[0044] It is also known that the degradation of melanosomes is induced by the activation of autophagic flux in keratinocytes (Pigment Cell Melanoma Res. 2020 May;33(3):403-415; Int. J. Mol. Sci 2021, 22,3995).

[0045] Beclin-1 protein and LC3-I / II protein are autophagy marker proteins known to be key factors involved in autophagy, and it is known that Beclin-1 protein and LC3-I / II protein increase when autophagy is activated.

[0046] As described above, the peptide of the present invention can exhibit skin whitening activity and preventive or therapeutic effects on hyperpigmentation diseases by inhibiting the activity of incorporation into melanosomes and the activity of degrading melanosomes.

[0047] 2. Composition for skin whitening and for preventing, treating or improving hyperpigmentation diseases

[0048] In another aspect of the present invention, provided is a composition for skin whitening, comprising a peptide comprising the amino acid sequence of SEQ ID NO: 1 as an active ingredient.

[0049] The peptide comprising the amino acid sequence of SEQ ID NO: 1 of the present invention has skin whitening activity through the mechanism as described above.

[0050] In another aspect of the present invention, the composition of the present invention may be a pharmaceutical composition for preventing or treating hyperpigmentation diseases, comprising a peptide comprising the amino acid sequence of SEQ ID NO: 1 as an active ingredient.

[0051] In one embodiment, the pharmaceutical composition comprising a peptide as an active ingredient inhibits the incorporation of melanosomes into keratinocytes or promotes the degradation of melanosomes.

[0052] In one embodiment, the pharmaceutical composition comprising the peptide inhibits the expression of one or more genes selected from the group consisting of a protease activated receptor 2 (PAR2) gene, a Toll-like receptor 3 (TLR3) gene, and a keratinocyte growth factor receptor (KGFR) gene in keratinocytes.

[0053] In one embodiment, the pharmaceutical composition comprising the peptide as an active ingredient upregulates the amount of Beclin-1 protein or LC3-I / II protein in keratinocytes.

[0054] As used herein, the term "hyperpigmentation disease" refers to a disease caused by an excessive increase in the amount of melanin in the skin. Thus, due to the excessive increase in the amount of melanin in the skin, the skin color becomes darker. The cause of the hyperpigmentation disease may be, for example, exposure to sunlight, skin inflammation such as acne (post-inflammatory hyperpigmentation), injury, hormonal imbalance or medication, but is not limited thereto.

[0055] In one embodiment, the hyperpigmentation disorder can be melasma, freckles, age spots, solar lentigo, or post-inflammatory hyperpigmentation of the skin. Post-inflammatory hyperpigmentation can be hyperpigmentation that occurs after injury, ultraviolet light, or inflammatory diseases of the skin, such as acne.

[0056] The pharmaceutical composition of the present invention may include a therapeutically effective amount of the peptide of the present invention comprising the amino acid of SEQ ID NO: 1.

[0057] The term "therapeutically effective amount" refers to an amount sufficient for the peptide to achieve its activity or efficacy, for example, an amount sufficient to achieve the efficacy of treating or preventing hyperpigmentation diseases, wherein the peptide is the active ingredient of the pharmaceutical composition of the present invention.

[0058] As used herein, the term "prevent" or "preventing" refers to reducing the risk of developing a disease or condition, and refers to all actions that inhibit or delay the onset of a disease by preventing the progression of the disease or one or more of its clinical symptoms.

[0059] As used herein, the term "treat" refers to ameliorating a disease or condition and includes all actions that ameliorate or beneficially alter the symptoms of a disease by arresting or reducing the progression of the disease or one or more of its clinical symptoms.

[0060] In the present invention, the prevention or treatment of hyperpigmentation diseases may be the removal of the cause of hyperpigmentation in the skin or the inhibition of the development of hyperpigmentation, and specifically, it may be the inhibition of melanosome incorporation into keratinocytes or the promotion of melanosome degradation.

[0061] The pharmaceutical composition of the present invention may include a pharmaceutically acceptable carrier.

[0062] Pharmaceutically acceptable carriers commonly used in formulations include lactose, dextrose, sucrose, sorbitol, mannitol, starch, gum arabic, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinyl pyrrolidone, cellulose, water, syrup, methylcellulose, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate, mineral oil, etc., but the carrier is not limited thereto.

[0063] In addition to the above ingredients, the pharmaceutical composition of the present invention may further include lubricants, wetting agents, sweeteners, flavoring agents, emulsifiers, suspending agents, preservatives, etc., but the ingredients are not limited thereto.

[0064] Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington: The Science and Practice of Pharmacy, (19th ed., 1995, Williams & Wilkins).

[0065] The pharmaceutical composition of the present invention can be administered by any route suitable for treating hyperpigmentation diseases, such as oral or parenteral administration. In the case of parenteral administration, the composition can be administered by intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, topical application, transdermal administration, etc. Since the pharmaceutical composition of the present invention has the activity of preventing or treating hyperpigmentation diseases, it is desirable to apply the composition topically, for example, by applying it to the skin.

[0066] The dosage of the pharmaceutical composition can be 0.0001 μg to 100 mg, 0.001 μg to 100 mg, 0.01 μg to 100 mg, 0.1 μg to 100 mg, or 1.0 μg to 1000 mg per day, but is not limited thereto. The pharmaceutical composition can be prescribed in various ways depending on factors such as formulation method, administration method, patient's age, weight, sex, pathological condition, food, administration time, administration route, excretion rate, and reaction sensitivity.

[0067] The pharmaceutical composition of the present invention can be prepared in unit dosage form by using a pharmaceutically acceptable carrier and / or excipient, according to a method that can be easily implemented by a person skilled in the art, or the pharmaceutical composition can be prepared by placing it in a multi-dose container. In particular, the formulation can be in the form of a solution, suspension or emulsion in an oil or aqueous medium, or can be in the form of an extract, powder, granule, tablet or capsule, and can additionally include a dispersant or stabilizer.

[0068] The pharmaceutical composition of the present invention can be an external skin preparation. An external skin preparation is a preparation that can be used by applying it to the outside of the skin. When the pharmaceutical composition of the present invention is used as an external skin preparation, it can be applied to skin areas where excessive pigmentation occurs. The external skin preparation can be a cream, a gel, an ointment, a skin emulsifier, a skin suspension, a transdermal delivery patch, a bandage including a drug, a lotion, or a combination thereof. The external skin preparation can be appropriately mixed with ingredients commonly used in external skin preparations (such as cosmetics and drugs) as needed, such as aqueous ingredients, oil-based ingredients, powder ingredients, alcohol, moisturizers, thickeners, ultraviolet absorbers, whitening agents, preservatives, antioxidants, surfactants, fragrances, colorants, various skin nutrients, or a combination thereof. The skin external preparation can be appropriately mixed with the following: metal chelators (such as disodium edetate, trisodium edetate, sodium citrate, sodium polyphosphate, sodium metaphosphate, gluconic acid, etc.), caffeine, tannin, licorice extract, glabridin, various herbs, tocopherol acetate, glycyrrhizic acid, drugs (such as tranexamic acid and its derivatives or salts, etc.), vitamin C, magnesium ascorbyl phosphate, ascorbyl glucoside, arbutin, kojic acid and sugars (such as glucose, fructose, trehalose, etc.), etc.

[0069] In another aspect of the present invention, the composition of the present invention provides a cosmetic composition for skin whitening, which comprises a peptide comprising the amino acid sequence of SEQ ID NO: 1 as an active ingredient.

[0070] In one embodiment, the cosmetic composition including the peptide of the present invention as an active ingredient inhibits the incorporation of melanosomes into keratinocytes or promotes the degradation of melanosomes.

[0071] In one embodiment, the cosmetic composition comprising the peptide of the present invention as an active ingredient inhibits the expression of one or more genes selected from the group consisting of protease activated receptor 2 (PAR2) gene, Toll-like receptor 3 (TLR3) gene and keratinocyte growth factor receptor (KGFR) gene.

[0072] In one embodiment, the cosmetic composition comprising the peptide of the present invention as an active ingredient upregulates the amount of Beclin-1 protein or LC3-I / II protein in keratinocytes.

[0073] The cosmetic composition can be prepared in any formulation commonly prepared in the art to which the present invention belongs, and can be an external skin preparation. For example, the cosmetic composition can be formulated as a solution, a suspension, an emulsion, a paste, a gel, a cream, a lotion, a powder, a soap, a surfactant-containing cleanser, an oil, a foundation, an emulsion foundation, a wax foundation, a spray, etc., but is not limited thereto.

[0074] The cosmetic composition can be prepared in various forms, such as solutions (e.g., softening lotions, nourishing lotions, nourishing creams, massage creams, essences, eye creams, cleansing creams, cleansing foams, cleansing waters, packs, sprays, powders, hair tonics, hair creams, hair lotions, hair shampoos, hair rinses, hair conditioners, hair sprays, hair aerosols, pomades, gels, etc.), sol-gels, emulsions, oils, waxes, aerosols, etc., but are not limited thereto.

[0075] The cosmetic composition of the present invention may include other additives such as excipients and carriers, and may apply and mix common ingredients mixed in general skin cosmetics as needed.

[0076] When the formulation of the cosmetic composition is a paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, tragacanth, cellulose derivatives, polyethylene glycol, silicone, bentonite, silicon dioxide, talc, zinc oxide and the like may be used as carrier components.

[0077] When the formulation of the cosmetic composition is a powder or a spray, lactose, talc, silicon dioxide, aluminum hydroxide, calcium silicate or polyamide powder can be used as a carrier component, and in particular, when the cosmetic composition is a spray, a propellant such as chlorofluorocarbon, propane / butane or dimethyl ether can be further included, but is not limited thereto.

[0078] When the cosmetic composition is in the form of a solution or emulsion, a solvent, solubilizer or emulsifier may be used as a carrier component, such as water, ethanol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol oil, aliphatic glycerol esters, polyethylene glycol, sorbitan fatty acid esters, etc.

[0079] When the preparation of the cosmetic composition is a suspension, liquid diluents (such as water, ethanol and propylene glycol), suspending agents (such as ethoxylated isostearyl alcohol, polyoxyethyl sorbitan esters, polyoxyethylene sorbitan esters, etc.), microcrystalline cellulose, aluminum metahydroxide, bentonite, agar, tragacanth gum, etc. can be used as carrier components.

[0080] When the formulation of the cosmetic composition is a surfactant-containing cleanser, fatty alcohol sulfates, fatty alcohol ether sulfates, sulfosuccinic acid monoesters, isethionates, imidazoline derivatives, methyl taurine, sarcosinates, fatty acid amide ether sulfates, alkylamide betaines, fatty alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives, ethoxylated glyceryl fatty acid esters, etc. can be used as carrier components.

[0081] When the formulation of the cosmetic composition is a shampoo, the peptide of the present invention can be mixed with basic ingredients for forming shampoo, such as thickeners, surfactants, viscosity regulators, moisturizers, pH regulators, preservatives, and essential oils. CDE can be used as a thickener; LES (anionic surfactant) and coco betaine (amphoteric surfactant) can be used as surfactants; polyquaternium salts (polyquarter) can be used as viscosity regulators; glycerin can be used as a moisturizer; and citric acid and sodium hydroxide can be used as pH regulators. Grapefruit extracts and the like can be used as preservatives; in addition, essential oils of cedar, mint, and rosemary, as well as silk amino acid, amyl alcohol, or vitamin E can be added.

[0082] In addition to the peptide of the present invention and the carrier component, the components included in the cosmetic composition as active ingredients may include auxiliary components commonly used in cosmetic compositions, such as antioxidants, stabilizers, solubilizers, vitamins, pigments and fragrances, but are not limited thereto.

[0083] The peptide of the present invention can be included in the above-mentioned composition, pharmaceutical composition or cosmetic composition at a concentration of 0.01 μM to 1000 μM, and specifically, the peptide of the present invention can be included in the above-mentioned composition, pharmaceutical composition or cosmetic composition at a concentration of 0.01 μM to 1000 μM; 0.05 μM to 800 μM, 0.05 μM to 700 μM, 0.05 μM to 600 μM, 0.05 μM to 500 μM, 0.05 μM to 300 μM or 0.05 μM to 200 μM; 0.1 μM to 800 μM, 0.1 μM to 700 μM, 0.1 μM to 600 μM, Concentrations of 0.1 μM to 500 μM, 0.1 μM to 300 μM or 0.1 μM to 200 μM; 1 μM to 800 μM, 1 μM to 700 μM, 1 μM to 600 μM, 1 μM to 500 μM, 1 μM to 300 μM or 1 μM to 200 μM; or 5 μM to 800 μM, 5 μM to 700 μM, 5 μM to 600 μM, 5 μM to 500 μM, 5 μM to 300 μM or 5 μM to 200 μM are included in the above-mentioned composition, pharmaceutical composition or cosmetic composition, but are not limited thereto.

[0084] 3. Uses of the peptides of the present invention

[0085] In another aspect of the present invention, a peptide comprising the amino acid sequence of SEQ ID NO: 1 is provided for use in skin whitening or for preventing, treating or ameliorating hyperpigmentation diseases.

[0086] In another aspect of the present invention, a method for skin whitening is provided, which comprises administering a peptide comprising the amino acid sequence of SEQ ID NO: 1 or a composition comprising the peptide to a subject in need of skin whitening.

[0087] In another aspect of the present invention, provided is a method for preventing or treating a hyperpigmentation disease, comprising administering a peptide comprising the amino acid sequence of SEQ ID NO: 1 or a composition comprising the peptide to a subject in need of prevention or treatment of a hyperpigmentation disease.

[0088] In another aspect of the present invention, provided is use of a peptide comprising the amino acid sequence of SEQ ID NO: 1 in the preparation of a medicament for preventing or treating hyperpigmentation diseases.

[0089] In another aspect of the present invention, provided is use of a peptide comprising the amino acid sequence of SEQ ID NO: 1 in preparing a cosmetic for skin whitening.

[0090] Beneficial Effects

[0091] The peptide of the present invention exhibits skin whitening activity by inhibiting the incorporation of melanosomes into keratinocytes and promoting the degradation of melanosomes. Therefore, the peptide of the present invention can be used as an active ingredient in a drug for treating or preventing hyperpigmentation diseases caused by the deposition of excessive melanosomes, or as an active ingredient in cosmetics for skin whitening.

[0092] However, the effects of the present invention are not limited to the above-mentioned effects, and other effects not mentioned will be clearly understood by those skilled in the art from the following description. BRIEF DESCRIPTION OF THE DRAWINGS

[0093] Figure 1 The results of melanin content analysis are shown, demonstrating that the peptides of the invention inhibit melanosome incorporation (phagocytosis) in HaCaT cells.

[0094] Figure 2a The results of PCR analysis are shown, indicating that the peptide of the present invention reduces the mRNA level of PAR2 gene, which is a receptor involved in melanosome phagocytosis, in HaCaT cells compared with the control group.

[0095] Figure 2b Shows the description Figure 2a A graph of the measured values ​​of the electrophoretic band density.

[0096] Figure 3a The results of PCR analysis are shown, indicating that the peptide of the present invention reduces the mRNA level of TLR3 gene, which is a receptor associated with melanosome phagocytosis, in HaCaT cells.

[0097] Figure 3b Shows the description Figure 3a A graph of the measured values ​​of the electrophoretic band density.

[0098] Figure 4a The results of PCR analysis are shown, demonstrating that the peptide of the present invention reduces the mRNA level of the KGFR gene, which is a receptor associated with melanophagocytosis, in HaCaT cells.

[0099] Figure 4b Shows the description Figure 4a A graph of the band density measured by electrophoresis.

[0100] Figure 5 The results of melanosome content analysis are shown, demonstrating that the peptides of the present invention induce melanosome degradation in HaCaT cells.

[0101] Figure 6aThe results of Western blot analysis are shown, illustrating that the amounts of Beclin-1 and LC3-I / II proteins, ie, autophagy-related marker proteins, were increased in HaCaT cells by treatment with the peptides of the present invention compared to the negative control (CON).

[0102] Figure 6b and Figure 6c Shows Figure 6a Figure 2. Graph of the measured values ​​of the band density shown in the PAGE gel. DETAILED DESCRIPTION

[0103] Hereinafter, the present invention will be described in detail. However, the following examples specifically illustrate the present invention, and the contents of the present invention are not limited to the following examples.

[0104] Preparation Example 1: Preparation of peptide

[0105] Peptides having the amino acid sequence of SEQ ID NO: 1 shown in Table 1 below were synthesized using an automatic peptide synthesizer (Milligen 9050, Millipore, USA), and then purified using C18 reverse phase high performance liquid chromatography (HPLC) (Waters Associates, USA). The column used was ACQUITY UPLC BEH300 C18 (2.1 mm×100 mm, 1.7 μm, Waters Co, USA).

[0106] [Table 1]

[0107]

[0108] The efficacy of the prepared peptide of SEQ ID NO: 1 was evaluated by the following experiment.

[0109] Experimental Example 1: Incorporation analysis via phagocytosis of melanosomes

[0110] It was confirmed whether the peptide of SEQ ID NO: 1 prepared in Preparation Example 1 inhibits the incorporation of melanosomes by phagocytosis in keratinocytes.

[0111] Human keratinocyte cell line (HaCaT cells) was cultured at 3 × 10 5The cells were seeded at a density of 100 μg / well in a 6-well cell culture plate and cultured in Dulbecco's modified Eagle's medium (DMEM) containing 10% FBS for 24 hours. Then, the cells were washed once with serum-free DMEM. A solution of the peptide of SEQ ID NO: 1 of Preparation Example 1 was added to 3 mL of serum-free DMEM medium to prepare peptide solutions at concentrations of 50 μM, 100 μM, and 200 μM, and the solutions were each added to the cultured cells. In particular, the negative control group (CON, (+)) was an untreated group, and the positive control group was treated with 500 nM of 6-amino-1-[4-(3-methyl-1-oxobutyl)-1-piperazinyl]-1-hexanone hydrochloride (ENMD-1068, Sigma-Aldrich). Then, the cells were incubated at 37°C in CO 2 After culturing them in an incubator for 24 hours, melanosomes (10 μg / mL) isolated from human epidermal melanocyte-darkly pigmented donors (HEM-DP) were additionally added to the cultured cells excluding the negative control (CON) group. Thereafter, the cells were incubated in a CO 2 The cells were cultured in an incubator at 37°C for 48 hours, washed three times with PBS, treated with 1X TE for 10 minutes, and the HaCaT cells were collected. The HaCaT cells were recovered using a centrifuge, dissolved in 1M NaOH, and distributed into a 96-well plate. The melanin content was measured by absorbance at 490 nm using an ELISA reader.

[0112] As a result of the experiment, Figure 1 As can be seen from the results of the melanin content analysis shown in , it was confirmed that the incorporation of melanosomes into HaCaT cells by melanosome phagocytosis was inhibited by the peptide of SEQ ID NO: 1 of the present invention.

[0113] Experimental Example 2: Analysis of the expression of PAR2 gene associated with melanosome phagocytosis

[0114] An experiment was conducted on the effect of the peptide of SEQ ID NO: 1 prepared in Preparation Example 1 on the expression of protease-activated receptor 2 (PAR2), a gene associated with melanosome phagocytosis, in keratinocytes. PAR2 is a gene associated with melanosome phagocytosis.

[0115] HaCaT cells were cultured at 3 × 10 5The cells were seeded at a density of 100 μM / well in a 6-well plate and cultured in a DMEM medium containing 10% FBS for 24 hours. Then, the cells were washed once with a serum-free DMEM medium. A solution of the peptide of SEQ ID NO.1 of Preparation Example 1 was added to 3 mL of a serum-free DMEM medium to prepare peptide solutions having concentrations of 10 μM, 50 μM, 100 μM and 200 μM, respectively, and the solutions were each distributed into the cells. In particular, the negative control group (CON, (+)) was an untreated group, and the positive control group was treated with 0.02X and 0.05X protease inhibitors (PI) (cOmplete TM Then, the cells were incubated at 37°C in CO 2 After 24 hours of incubation in the incubator, 4 units of trypsin were added to the cultured cells excluding the negative control (CON) group. Thereafter, the cells were incubated in CO 2 Incubate at 37°C for 16 hours in an incubator, wash twice with PBS, and separate RNA using easy blue (iNtRON, Catalog No.: 17061, South Korea). After quantifying the amount of isolated RNA, 2000 ng RNA was dispensed per tube, and cDNA was synthesized using a cDNA synthesis kit (Enzynomics, Catalog No.: RT200, South Korea). PCR was performed using primers targeting the PAR2 gene shown in Table 2 and a PCR kit (Enzynomics, Catalog No.: P581T, South Korea). Next, the PCR product was electrophoresed on a 1.2% agarose gel, and the bands were detected and analyzed using a Bio-Rad gel image system.

[0116] [Table 2]

[0117]

[0118] As a result of the experiment, Figure 2a and 2b As shown, it was confirmed that the peptide of SEQ ID NO: 1 of the present invention reduced the mRNA level of PAR2 gene, which is a receptor involved in melanosome phagocytosis (incorporation), in HaCaT cells compared with the control group. These results confirm that the peptide of SEQ ID NO: 1 inhibits the incorporation of melanosomes.

[0119] Experimental Example 3: Expression analysis of TLR3 gene associated with melanosome phagocytosis

[0120] An experiment was conducted on the effect of the peptide of SEQ ID NO: 1 prepared in Preparation Example 1 on the expression of Toll-like receptor 3 (TLR3), a gene associated with melanosome phagocytosis, in keratinocytes.

[0121] HaCaT cells were cultured at 3 × 10 5 The cells were seeded at a density of 100 cells / well in a 6-well plate and cultured in DMEM medium containing 10% FBS for 24 hours. Then, the cells were washed once with serum-free DMEM medium, and then 2 mL of serum-free DMEM medium was added to the cells. The cells were then incubated in CO 2 The cells were incubated at 37°C in an incubator for 24 hours. The peptide of SEQ ID NO: 1 of Preparation Example 1 was added to 3 mL of a DMEM medium containing 2% serum to prepare peptide solutions at concentrations of 10 μM, 50 μM, 100 μM, and 200 μM, and each of these solutions was added to the cells. After 30 minutes, 30 μg / mL of polyinosinic acid (poly(I:C)) was additionally added. Subsequently, the cells were incubated at 37°C for 24 hours. 2 Incubate at 37°C for 24 hours in an incubator, wash twice with PBS, and separate RNA using easy blue (iNtRON, Catalog No.: 17061, South Korea). After quantifying the amount of isolated RNA, 2000 ng RNA was allocated to each tube, and cDNA was synthesized using a cDNA synthesis kit (Enzynomics, Catalog No.: RT200, South Korea). PCR was performed using primers targeting the TLR3 gene in Table 3 and a PCR kit (Enzynomics, Catalog No.: P581T, South Korea). Then, the PCR product was electrophoresed on a 1.2% agarose gel, and the bands were detected and analyzed using a Bio-Rad gel image system.

[0122] [Table 3]

[0123]

[0124] As a result of the experiment, Figure 3a and 3b As shown, it was confirmed that the peptide of SEQ ID NO: 1 reduced the mRNA level of the gene of TLR3, which is a receptor involved in melanosome phagocytosis (incorporation), in HaCaT cells compared with the control group. These results confirmed that the peptide of SEQ ID NO: 1 inhibited the incorporation of melanosomes.

[0125] Experimental Example 4: Expression analysis of KGFR gene associated with melanosome phagocytosis

[0126] An experiment was conducted on the effect of the peptide of SEQ ID NO: 1 prepared in Preparation Example 1 on the expression of keratinocyte growth factor receptor (KGFR), which is a melanosome phagocytosis-related gene, in keratinocytes.

[0127] HaCaT cells were cultured at 3 × 10 5 The cells were seeded at a density of 100 cells / well in a 6-well plate and cultured in DMEM medium containing 10% FBS for 24 hours. Then, the cells were washed once with serum-free DMEM medium, and then 2 mL of serum-free DMEM medium was added to the cells. The cells were then incubated in CO 2 The cells were cultured in an incubator at 37° C. for 24 hours. The peptide of SEQ ID NO: 1 of Preparation Example 1 was added to 3 mL of DMEM medium containing 2% serum to prepare peptide solutions with concentrations of 10 μM, 50 μM, 100 μM, and 200 μM, and each of these solutions was added to the cells and the cells were incubated in CO 2 Incubator at 37 ° C for 6 hours. Subsequently, the cells were washed twice with PBS, and RNA was isolated using easy blue (iNtRON, Catalog No.: 17061, South Korea). After quantifying the amount of isolated RNA, RNA was distributed in an amount of 2000 ng per tube, and cDNA was synthesized using a cDNA synthesis kit (Enzynomics, Catalog No.: RT200, South Korea). PCR was performed using primers targeting the KGFR gene in Table 4 and a PCR kit (Enzynomics, Catalog No.: P581T, South Korea). Then, the PCR product was electrophoresed on a 1.2% agarose gel, and the bands were detected and analyzed using a Bio-Rad gel image system.

[0128] [Table 4]

[0129]

[0130] As a result of the experiment, Figure 4a and 4b As shown, it was confirmed that the peptide of SEQ ID NO: 1 reduced the mRNA level of the gene of KGFR, which is a receptor involved in melanosome phagocytosis (incorporation), in HaCaT cells compared with the control group. These results confirmed that the peptide of SEQ ID NO: 1 inhibited the incorporation of melanosomes.

[0131] Experimental Example 5: Melanosome degradation analysis

[0132] An analysis was performed as to whether the peptide of SEQ ID NO: 1 prepared in Preparation Example 1 degrades melanosomes in keratinocytes.

[0133] HaCaT cells were cultured at 3 × 105 The cells were seeded at a density of 100 μg / mL in a 6-well plate and cultured in a DMEM medium containing 10% FBS for 24 hours. Then, melanosomes isolated from human epidermal melanocytes-dark pigment donors (HEM-DP) were added to 2 mL of a DMEM medium containing 2% serum to prepare a melanosome solution with a concentration of 10 μg / mL, which was dispensed into the cells and the cells were incubated at 37°C under CO 2 Incubate in an incubator for 48 hours.

[0134] Subsequently, the melanosomes separated from the HEM-DP were added to 3 mL of a DMEM medium including 2% serum to prepare a melanosome solution at a concentration of 10 μg / mL, and the peptide of SEQ ID NO: 1 of Preparation Example 1 was added to 3 mL of a DMEM medium including 2% serum to prepare peptide solutions at concentrations of 10 μM, 50 μM, 100 μM, and 200 μM, and the melanosome solution and the peptide solution were additionally dispensed into the cells. In particular, the negative control group (CON) was an untreated group, wherein the negative control group (+) was treated with the melanosome solution alone, and the positive control group included a group (Rapa) treated with the melanosome solution and 100 nM and 200 nM rapamycin, and a group (RSV) treated with the melanosome solution and 200 μM and 500 μM resveratrol. Then, the cells were incubated in CO 2 The cells were cultured in an incubator at 37°C for 72 hours, washed three times with PBS, and then treated with 1X TE for 10 minutes, and the HaCaT cells were collected. The HaCaT cells were recovered using a centrifuge, dissolved in 1M NaOH, and distributed into a 96-well plate. The melanin content was measured by absorbance at 490 nm using an ELISA reader.

[0135] As a result of the experiment, Figure 5 As can be seen from the changes in the melanosome content in the cells, it was confirmed that treatment with the peptide of SEQ ID NO: 1 induced the degradation of melanosomes in HaCaT cells.

[0136] Experimental Example 6: Activation analysis of autophagic flux

[0137] The effect of the peptide of SEQ ID NO: 1 prepared in Preparation Example 1 on the activation of autophagic flux in human keratinocytes in terms of autophagic markers (ie, Beclin-1, LC3-I, and LC3-II proteins) was confirmed by Western blot analysis.

[0138] HaCaT cells were cultured at 3 × 10 5The cells were seeded at a density of 100 μM / well in a 6-well cell culture plate and cultured in a DMEM medium containing 10% FBS for 24 hours. Then, the cells were washed once with a serum-free DMEM medium, and then the peptide of SEQ ID NO: 1 of Preparation Example 1 was added to 1 mL of a serum-free DMEM medium to prepare peptide solutions having concentrations of 50 μM, 100 μM, and 200 μM, respectively, and dispensed into the cells. In particular, the negative control group (CON) was an untreated group, and the positive control group included a group treated with 100 nM and 200 nM rapamycin (Rapa) and a group treated with 50 μM and 100 μM resveratrol (RSV). Subsequently, the cells were incubated in CO 2 Incubate at 37°C for 3 hours in an incubator, wash twice with PBS, and lyse the cells by adding cell lysis buffer (Whole Cell Extraction Kit, Millipore, USA). Prepare the sample by treating with 5X sample buffer, and perform SDS-PAGE using 8% SDS-PAGE gel. Proteins separated by SDS-PAGE were transferred to PVDF membrane. Blocking was performed using 5% skim milk at room temperature for 30 minutes. The following primary antibodies were diluted 1:1000 in 5% BSA and reacted with the membrane for 2 hours. Primary antibodies: anti-Beclin-1 antibody (Cell signaling, cat. no.: #3495, USA) and anti-LC3-I / II antibody (Cell signaling, cat. no.: #4108, USA). Subsequently, the antibody was washed three times with 0.1% PBS-T (PBS with 0.1% Tween-20) for 10 minutes each time, and goat anti-rabbit IgG (Jackson Immune Research, Catalog No.: 111-035-033, USA) was diluted 1:3000 in 5% skim milk and reacted with the membrane for 1 hour. Subsequently, the resultant was washed three times with 0.1% PBS-T (PBS with 0.1% Tween-20) for 10 minutes each time, and detected on the membrane using ECL solution (GE Healthcare, Catalog No.: RPN2232, USA).

[0139] As a result of the experiment, Figures 6a to 6c As can be seen in the figure, by treating with the peptide of SEQ ID NO: 1, it was confirmed that the amount of Beclin-1 protein and LC3-I / II protein in HaCaT cells was increased compared with the negative control (CON). These results indicate that the peptide of SEQ ID NO: 1 promotes the degradation of melanosomes by activating the autophagic flux in HaCaT cells.

[0140] Preparation Example 2: Preparation of pharmaceutical composition

[0141] 2-1. Preparation of ointment

[0142] Peptide 5g of the present invention

[0143] Cetyl palmitate 20g

[0144] Cetyl alcohol 40g

[0145] Octadecanol 40g

[0146] Myristane isopropyl 80g

[0147] Polysorbate 60g

[0148] Propyl paraoxybenzoate 1g

[0149] Methyl paraoxybenzoate 1g

[0150] Phosphoric acid and purified water in sufficient quantities

[0151] The ointment is prepared by incorporating the above ingredients in the specified contents according to a conventional ointment preparation method.

[0152] 2-2. Preparation of powder

[0153] Peptide 2g of the present invention

[0154] Lactose 1g

[0155] The powder is prepared by mixing the above ingredients and then filling the mixture into airtight bags.

[0156] 2-3. Tablet preparation

[0157] Peptide of the present invention 100 mg

[0158] Corn starch 100mg

[0159] Lactose 100mg

[0160] Magnesium stearate 2mg

[0161] Tablets are prepared by mixing the above ingredients and then tableting the mixture according to a conventional tablet preparation method.

[0162] 2-4. Capsule preparation

[0163] Peptide of the present invention 100 mg

[0164] Corn starch 100mg

[0165] Lactose 100mg

[0166] Magnesium stearate 2mg

[0167] Capsules are prepared by mixing the above ingredients and then filling the mixture into gelatin capsules according to a conventional capsule preparation method.

[0168] 2-5. Preparation of pills

[0169] Peptide 1g of the present invention

[0170] Lactose 1.5g

[0171] Glycerin 1g

[0172] Xylitol 0.5g

[0173] After mixing the above ingredients, pellets were prepared in an amount of 4 g per pellet according to a conventional method.

[0174] Preparation Example 3: Preparation of cosmetic composition

[0175] 3-1. Preparation of cream

[0176] 4.6 parts by weight of the peptide of the present invention

[0177] Cetostearyl alcohol 2.8 parts by weight

[0178] Beeswax 2.6 parts by weight

[0179] 1.4 parts by weight of stearic acid

[0180] 2 parts by weight of lipophilic glyceryl monostearate

[0181] PEG-100 stearate 1 part by weight

[0182] Sorbitan sesquioleate 1.4 parts by weight

[0183] 4 parts by weight of jojoba oil

[0184] Squalane 3.8 parts by weight

[0185] Polysorbate 60 1.1 parts by weight

[0186] Macadamia oil 2 parts by weight

[0187] Tocopherol acetate 0.2 parts by weight

[0188] Methyl polysiloxane 0.4 parts by weight

[0189] Ethyl p-hydroxybenzoate 0.1 parts by weight

[0190] Propyl p-hydroxybenzoate 0.1 parts by weight

[0191] Euxyl K-400 0.1 parts by weight

[0192] 1.3-Butene diol 7 parts by weight

[0193] Methyl p-hydroxybenzoate 0.05 parts by weight

[0194] Glycerol 6 parts by weight

[0195] D-Panthenol 0.2 parts by weight

[0196] Triethanolamine 0.2 parts by weight

[0197] pt 41891 0.2 parts by weight

[0198] pH 2 O 46.05 parts by weight

[0199] 3-2. Preparation of lotion

[0200] 3.5 parts by weight of the peptide of the present invention

[0201] Cetostearyl alcohol 1.6 parts by weight

[0202] 1.4 parts by weight of stearic acid

[0203] 1.8 parts by weight of lipophilic glyceryl monostearate

[0204] PEG-100 stearate 2.6 parts by weight

[0205] Sorbitan sesquioleate 0.6 parts by weight

[0206] Squalene 4.8 parts by weight

[0207] Macadamia oil 2 parts by weight

[0208] 2 parts by weight of jojoba oil

[0209] Tocopherol acetate 0.4 parts by weight

[0210] Methyl polysiloxane 0.2 parts by weight

[0211] Ethyl p-hydroxybenzoate 0.1 parts by weight

[0212] Propyl p-hydroxybenzoate 0.1 parts by weight

[0213] 1. 4 parts by weight of 3-butene diol

[0214] Methyl p-hydroxybenzoate 0.1 parts by weight

[0215] Xanthan gum 0.1 parts by weight

[0216] 4 parts by weight of glycerol

[0217] D-Panthenol 0.15 parts by weight

[0218] Allantoin 0.1 parts by weight

[0219] Calcium carbonate (2% aqueous solution) 4 parts by weight

[0220] Triethanolamine 0.15 parts by weight

[0221] 3 parts by weight of ethanol

[0222] pt 41891 0.1 parts by weight

[0223] pH 2 O 48.3 parts by weight

[0224] 3-3. Preparation of softening lotion

[0225] Peptide of the present invention 0.2 wt%

[0226] Ethanol 10.0 wt%

[0227] Polyoxyethylene sorbitan polylaurate 1.0 wt%

[0228] Methyl paraoxybenzoate 0.2 wt%

[0229] Glycerol 5.0 wt%

[0230] 1. 3-Butanediol 6.0wt%

[0231] Enough fragrance

[0232] Sufficient pigment

[0233] Sufficient amount of purified water

[0234] Total 100

[0235] 3-4. Preparation of nourishing lotion

[0236] Peptide of the present invention 0.1 wt%

[0237] Vaseline 2.0 wt%

[0238] Sorbitan sesquioleate 0.8 wt%

[0239] Polyoxyethylene oleyl ethyl 1.2 wt%

[0240] Methylparaben Adequate

[0241] Propylene glycol 5.0 wt%

[0242] Ethanol 3.2 wt%

[0243] Carboxyvinyl polymer 18.0 wt%

[0244] Sufficient pigment

[0245] Enough fragrance

[0246] Sufficient amount of purified water

[0247] Total 100

[0248] 3-5. Preparation of Essence

[0249] Peptide of the present invention 5.0 wt%

[0250] Propylene glycol 10.0 wt%

[0251] Glycerol 10.0 wt%

[0252] Sodium hyaluronate (1%) aqueous solution 5.0 wt%

[0253] Ethanol 3.2 wt%

[0254] Polyoxyethylene hydrogenated castor oil 1.0 wt%

[0255] Methyl paraben 0.1 wt%

[0256] Enough fragrance

[0257] Sufficient amount of purified water

[0258] Total 100

[0259] 3-6. Preparation of membrane (pack)

[0260] Peptide of the present invention 0.5 wt%

[0261] Glycerol 5.0 wt%

[0262] Propylene glycol 4.0 wt%

[0263] Polyvinyl alcohol 15.0 wt%

[0264] Ethanol 8.0 wt%

[0265] Polyoxyethylene oleyl ethyl 1.0 wt%

[0266] Methyl paraoxybenzoate 0.2 wt%

[0267] Enough fragrance

[0268] Sufficient pigment

[0269] Sufficient amount of purified water

[0270] Total 100

[0271] The above-mentioned composition ratio is a mixture of suitable ingredients in a preferred embodiment, but the ingredients or the mixing ratio thereof may be arbitrarily modified according to regional or ethnic preferences (eg, demand segment, demand country, and intended use).

[0272] Hereinabove, representative embodiments of the present application have been described through exemplary embodiments, but the scope of the present application is not limited to the above-mentioned specific embodiments, and those skilled in the art will be able to make appropriate modifications within the scope described in the claims of the present application.

Claims

1. A peptide comprising the amino acid sequence of SEQ ID NO:

1.

2. A composition for skin whitening, comprising the peptide according to claim 1 as an active ingredient.

3. A pharmaceutical composition for preventing or treating hyperpigmentation diseases, comprising the peptide according to claim 1 as an active ingredient.

4. The pharmaceutical composition of claim 3, wherein in keratinocytes, the peptide: (i) inhibiting melanosome incorporation; or (ii) Promote the degradation of melanosomes.

5. The pharmaceutical composition of claim 4, wherein in keratinocytes, the peptide: (i) inhibiting the expression of one or more genes selected from the group consisting of the protease activated receptor 2 (PAR2) gene, the Toll-like receptor 3 (TLR3) gene and the keratinocyte growth factor receptor (KGFR) gene; or (ii) Upregulation of Beclin-1 protein or LC3-I / II protein.

6. The pharmaceutical composition of claim 3, wherein the hyperpigmentation disease is a disease that occurs when the amount of melanin in the skin excessively increases.

7. The pharmaceutical composition of claim 3, wherein the hyperpigmentation disease is melasma, freckles, age spots, solar lentigo, or hyperpigmentation of the skin after inflammation.

8. The pharmaceutical composition of claim 3, wherein the pharmaceutical composition is a skin preparation for external use.

9. A cosmetic composition for skin whitening, comprising the peptide of claim 1 as an active ingredient.

10. The cosmetic composition of claim 9, wherein in keratinocytes, the peptide: (i) inhibiting melanosome incorporation; or (ii) Promote the degradation of melanosomes.

11. The cosmetic composition of claim 10, wherein in keratinocytes, the peptide: (i) inhibiting the expression of one or more genes selected from the group consisting of the protease activated receptor 2 (PAR2) gene, the Toll-like receptor 3 (TLR3) gene and the keratinocyte growth factor receptor (KGFR) gene; or (ii) Upregulation of Beclin-1 protein or LC3-I / II protein.

12. The cosmetic composition of claim 9, wherein the cosmetic composition is a skin preparation for external use.

13. The cosmetic composition of claim 12, wherein the cosmetic composition is at least one formulation selected from the group consisting of solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, surfactant-containing cleansers, oils, foundations, emulsion foundations, wax foundations, and sprays.

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