Peptides having skin-whitening activity and uses thereof

Abstract

The present application relates to a peptide having skin whitening activity and use thereof, and provides: a peptide consisting of the amino acid sequence of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3; a composition for skin whitening comprising the same; a cosmetic composition comprising the composition for skin whitening; and a pharmaceutical composition for preventing or treating hyperpigmentation diseases comprising the composition for skin whitening.

Description

[0001] This application is a divisional application of the same-named invention patent application 201980038477.X, filed on October 10, 2019. TECHNICAL FIELD

[0002] The present application relates to a peptide having skin whitening activity and uses thereof. BACKGROUND

[0003] In addition to the differences according to race, region, gender, and age, the factors determining skin color basically include chloasma, freckles, and tanning by exposure to ultraviolet rays, and also include overall pigmentation, acne, scars, keratin distribution, blood circulation, stress, and health status, etc. Among the factors, pigmentation is known as the main factor determining skin color.

[0004] The pigments affecting skin color include melanin, melanin-like substances, carotene, hemoglobin, and carotenoid, and the various colors of skin, hair, and eyes depend on these pigments. Among them, the most important pigment determining skin color is melanin, and specifically, skin color depends on the amount and distribution of melanin. Melanin is generated in cells called melanocytes under the epidermis, and is moved to the keratin surface by the metabolism of the skin and is exfoliated. Regardless of skin color, the number of melanocytes is almost the same, but skin color is different due to differences in the production, type, and distribution of melanin.

[0005] In the skin, tyrosine is converted into dihydroxyphenylalanine (DOPA) by an enzyme called tyrosinase in the human body, and through a series of oxidation processes, melanin, which is a black-brown polymer, is finally produced. When the production of melanin is excessive, it can cause excessive pigmentation of chloasma, freckles, and mottling, thereby causing an undesirable result in terms of cosmetics. Currently, as the number of people enjoying outdoor activities increases due to the increase in leisure population, the demand for preventing melanin pigmentation caused by ultraviolet rays is also increasing.

[0006] In this technical background, research on whitening agents that prevent excessive melanin production through mechanisms such as inhibition of melanocyte activity, inhibition of tyrosinase activity, etc. is being developed from various angles (Korean Patent Publication No. 10-2019-0109722), but it is still insignificant. SUMMARY

[0007] TECHNICAL PROBLEM

[0008] In one aspect, a peptide consisting of the amino acid sequence of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3 is provided.

[0009] Another aspect provides a composition for skin whitening, including a peptide consisting of the amino acid sequence of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3 as an effective ingredient.

[0010] Another aspect provides a cosmetic composition, including the composition for skin whitening.

[0011] Another aspect provides a pharmaceutical composition for preventing or treating a hypermelanosis disease, including the composition for skin whitening.

[0012] Other objects and advantages of the present application will be understood more clearly from the appended claims and drawings and the following detailed description. For matters not recited in the specification, what is well known and can be inferred by those skilled in the art or in a similar technical field is omitted from the description.

[0013] Technical Solution

[0014] The descriptions and embodiments disclosed in the present application can be applied to other descriptions and embodiments. In other words, all combinations of the various technical features disclosed in the present application belong to the scope of the present application. Also, the scope of the present application is not limited to the specific descriptions recited below.

[0015] One aspect provides a peptide consisting of the amino acid sequence of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3.

[0016] The term "peptide" used in the present specification can mean a linear molecule in which amino acid residues are combined with each other by peptide bonding. The peptide can be prepared according to a chemical synthesis method well known in the art, particularly, according to solid-phase synthesis techniques (Merrifield, J. Amer. Chem. Soc. 85:2149-54 (1963); Stewart, et al., Solid Phase Peptide Synthesis, 2nd Ed., Pierce Chem. Co.: Rockford, Ill (1984)) or liquid-phase synthesis techniques (U.S. Pat. No. 5,516,891). The present inventors have made efforts to develop a peptide having a biologically effective activity, and as a result, a peptide consisting of the amino acid sequence of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3 has been identified. Here, the biologically effective activity can mean any one or more of the properties selected from the group consisting of: (a) inhibiting the production of melanin; (b) inhibiting the activity of tyrosinase; and (c) inhibiting the expression of Rab27a, melanophilin, myosin VA, microphthalmia transcription factor, tyrosinase, or tyrosinase-related protein 1. Accordingly, the peptide is used for skin whitening, prevention or treatment of a hypermelanosis disease.

[0017] In order to obtain chemical stability, enhanced pharmacological properties (half-life, absorption, titration, potency, etc.), altered specificity (e.g., broad biological activity spectrum), reduced antigenicity, a protecting group can be bound to the N- or C-terminus of the peptide. In a specific embodiment, the N-terminus of the peptide is bound to any one protecting group selected from the group consisting of an acetyl group, a fluorenylmethoxycarbonyl group, a formyl group, a palmitoyl group, a myristyl group, a stearyl group, a butoxycarbonyl group, an allyloxycarbonyl group, and a polyethylene glycol (PEG); and / or the C-terminus of the peptide is bound to any one protecting group selected from the group consisting of an amino group (-NH2), a tertiary alkyl group, and an azide (-NHNH2). Also, the peptide can optionally further include a leader sequence, a tag, a labeled residue, a half-life, or an amino acid sequence manufactured for the specific purpose of improving peptide stability.

[0018] The term "stability" used in the present specification can mean not only the in vivo stability of the peptide protected from the attack of proteases in vivo, but also storage stability (e.g., room temperature storage stability).

[0019] Other aspects provide a cosmetic composition including a peptide including the amino acid sequence of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3 as an effective ingredient; a pharmaceutical composition including a peptide including the amino acid sequence of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3 as an effective ingredient; and a use of a peptide including the amino acid sequence of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3 in the preparation of a cosmetic or pharmaceutical composition or as a cosmetic or pharmaceutical composition.

[0020] Another aspect provides a composition for skin whitening including a peptide consisting of the amino acid sequence of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3 as an effective ingredient.

[0021] Among the terms or technical features raised in the description of the peptide, the same terms or technical features as those already raised are as described above.

[0022] The term "skin whitening" used in the present specification can be interpreted as not only inhibiting the synthesis of melanin to lighten the skin tone but also improving the excessive pigmentation of the skin due to ultraviolet rays, hormones, or heredity. Here, the excessive pigmentation of the skin can include freckles, chloasma, senile lentigo, brown spots, or age spots, but is not limited thereto.

[0023] The term "improving" used in the present specification can mean all actions of alleviating or at least reducing the degree of a parameter related to treatment such as a symptom of a state.

[0024] Although the existing functional peptides have effective biological activities, they have problems in that they cannot be effectively introduced into target tissues or cells or are eliminated in a short time in vivo due to the size of the peptides themselves. In contrast, the composition for whitening according to an embodiment includes a peptide consisting of 10 or less amino acids as an effective ingredient, and thus, the skin permeation rate or the like of the effective ingredient is very excellent, for example, when applied to the skin, an effective skin whitening effect can be obtained.

[0025] According to an embodiment, the peptide can significantly inhibit the generation of melanin and the activity of tyrosinase. Also, the peptide can inhibit the expression of melanin generation-related factors, i.e., Rab27a, melanophilin (Mlph), myosin VA, melanocyte inducing transcription factor (MITF), tyrosinase, or tyrosinase-related protein 1 (TRP1). Therefore, the peptide can be used as an effective ingredient of a cosmetic composition for skin whitening.

[0026] On the other hand, there is provided a cosmetic composition for skin whitening, which includes a peptide consisting of the amino acid sequence of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3 as an effective ingredient.

[0027] Among the terms or technical features raised in the description of the peptide, the same terms or technical features as those already raised are as described above.

[0028] The cosmetic composition can include a cosmetically effective amount of the peptide; and / or a cosmetically acceptable carrier, but is not limited thereto.

[0029] The term "cosmetically effective amount" used in the present specification means an amount sufficient to achieve the skin whitening efficacy of the cosmetic composition.

[0030] The weight ratio of the peptide and the cosmetically acceptable carrier can be, for example, 500: 1 to 1: 500, and as one example, the weight ratio can be 450: 1 to 1: 450, 400: 1 to 1: 400, 350: 1 to 1: 350, 300: 1 to 1: 300, 250: 1 to 1: 250, 200: 1 to 1: 200, 150: 1 to 1: 150, 100: 1 to 1: 100, 80: 1 to 1: 80, 60: 1 to 1: 60, 40: 1 to 1: 40, 20: 1 to 1: 20, 10: 1 to 1: 10, 8: 1 to 1: 8, 6: 1 to 1: 6, 4: 1 to 1: 4, or 2: 1 to 1: 2, but is not limited thereto.

[0031] The cosmetic composition can be prepared in any dosage form generally prepared in the art, for example, a solution, a suspension, an emulsion, a paste, a gel, a cream, a lotion, a powder, a soap, a detergent including a surfactant, an oil, a powder type foundation, an emulsion type foundation, a wax type foundation, and a hair spray, etc., but is not limited thereto. For example, the cosmetic composition can be prepared in the dosage form of a softening toner, a nourishing toner, a nourishing cream, a massage cream, an essence, an eye cream, a cleansing paste, a cleansing foam, a cleansing water, a pack, a hair spray, or a foundation.

[0032] When the dosage form of the cosmetic composition is a paste, a cream, or a gel, an animal oil, a vegetable oil, a wax, a paraffin, a starch, a tragacanth gum, a cellulose derivative, a polyethylene glycol, bentonite, silica, talc, or zinc oxide, etc. can be used as a carrier ingredient.

[0033] When the dosage form of the cosmetic composition is a powder or a hair spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, or a polyamine acid powder can be used as a carrier ingredient, and for example, when the cosmetic composition is a hair spray, a propellant such as chlorofluorocarbon, propane / butane, or dimethyl ether can be further included.

[0034] When the dosage form of the cosmetic composition is a solution or an emulsion, a solvent, a dissolving agent, or an emulsifying agent can be used as a carrier ingredient, and for example, water, ethanol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol oil, a fatty ester of glycerol, or a fatty acid ester of sorbitan can be included.

[0035] When the dosage form of the cosmetic composition is a suspension, a liquid diluent such as water, ethanol, or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, a polyoxyethylene sorbitol ester, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar, or tragacanth gum, etc. can be used as a carrier ingredient.

[0036] When the dosage form of the cosmetic composition is a detergent including a surfactant, a fatty alcohol sulfate, a fatty alcohol ether sulfate, a succinic monoester, an isethionate, an imidazoline derivative, a methyl ursodeoxycholate, a sarcosinate, a fatty amide ether sulfate, a chitosan, a fatty alcohol, a fatty acid glyceride, a fatty acid diethanolamide, a vegetable oil, a lanolin derivative, or an ethoxylated propylene glycol fatty acid ester, etc. can be used as the carrier ingredient.

[0037] In addition to the peptide as the effective ingredient and the carrier ingredient, the ingredients included in the cosmetic include the ingredients generally used in cosmetic compositions, for example, the conventional adjuvants such as antioxidants, stabilizers, dissolving agents, vitamins, pigments, and fragrances can be included.

[0038] The other aspect provides a skin whitening method including applying a cosmetic composition including a peptide consisting of the amino acid sequence of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3 as an effective ingredient on the skin of an individual.

[0039] Among the terms or technical features raised in the description regarding the cosmetic composition, the same terms as the terms or technical features already raised are as described above.

[0040] The terms "application", "administration", and "application" used in the present specification can be used interchangeably, and can mean to cause the localization of at least a part of the composition according to an embodiment to a desired site, or can mean to dispose the composition according to an embodiment in an individual according to an administration route.

[0041] The other aspect provides a pharmaceutical composition for preventing or treating a hyperpigmentation disease, including a peptide consisting of the amino acid sequence of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3 as an effective ingredient.

[0042] Among the terms or technical features raised in the description regarding the peptide, the same terms or technical features as the terms or technical features already raised are as described above.

[0043] In the present specification, the term "prevention" means all actions to inhibit or delay the occurrence of a disease by administering the composition.

[0044] In the present specification, the term "treatment" means any form of treatment that provides an individual having a disease or at risk of developing a disease with an effect including improvement of the state (e.g., one or more symptoms) of the individual, delay of disease progression, delay of symptom occurrence, or inactivation of symptom progression, etc. Therefore, the "treatment" and "prevention" do not mean the cure or complete removal of symptoms.

[0045] The "individual" refers to a subject in need of treatment of a disease, more specifically, a human or a non-human primate, a mouse, a dog, a cat, a horse, a cow, and the like mammal.

[0046] As a disease to be prevented or treated by the pharmaceutical composition, the "melanin hyperpigmentation disease" can be a disease in which melanin is excessively produced and deposited in the skin, for example, can be chloasma, freckle, senile lentigo, or solar lentigo, but is not limited thereto.

[0047] The pharmaceutical composition can include a pharmaceutically effective amount of the peptide; and / or a pharmaceutically acceptable carrier, but is not limited thereto.

[0048] The term "pharmaceutically effective amount" used in the present specification can refer to an amount sufficient to achieve the prophylactic or therapeutic efficacy of the pharmaceutical composition on the melanin hyperpigmentation disease.

[0049] The weight ratio of the peptide and the pharmaceutically acceptable carrier can be, for example, 500: 1 to 1: 500, as an example, the weight ratio can be 450: 1 to 1: 450, 400: 1 to 1: 400, 350: 1 to 1: 350, 300: 1 to 1: 300, 250: 1 to 1: 250, 200: 1 to 1: 200, 150: 1 to 1: 150, 100: 1 to 1: 100, 80: 1 to 1: 80, 60: 1 to 1: 60, 40: 1 to 1: 40, 20: 1 to 1: 20, 10: 1 to 1: 10, 8: 1 to 1: 8, 6: 1 to 1: 6, 4: 1 to 1: 4, or 2: 1 to 1: 2, but is not limited thereto.

[0050] The pharmaceutically acceptable carrier is generally used in the preparation of a dosage form, including lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, sugar syrup, methyl cellulose, hydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil, but is not limited thereto. Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington's Pharmaceutical Sciences (19th edition, 1995).

[0051] The pharmaceutical composition can further include a lubricant, a humectant, a sweetener, a flavoring agent, an emulsifier, a suspending agent, a preservative, and the like, in addition to the components, but is not limited thereto.

[0052] The pharmaceutical composition can be administered orally or parenterally, preferably can be administered parenterally, and when administered parenterally, can be administered by injection into muscle, intravenous injection, subcutaneous injection, intraperitoneal injection, topical application, transdermal application, etc., but is not limited thereto.

[0053] The administration amount of the pharmaceutical composition can be 0.0001 to 1000 μg, 0.001 to 1000 μg, 0.01 to 1000 μg, 0.1 to 1000 μg, 1.0 to 1000 μg per day, but is not limited thereto, and can be prescribed differently according to factors such as the formulation method, the administration method, the patient's age, weight, sex, condition, diet, administration time, administration route, excretion rate, and response sensitivity, etc.

[0054] The pharmaceutical composition is prepared by formulating using pharmaceutically acceptable carriers and / or excipients in a unit capacity form or in a large capacity container according to a method that can be easily performed by a person of ordinary skill in the art to which the present application pertains.

[0055] The dosage form 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 further include a dispersant and / or stabilizer.

[0056] Another aspect includes a method of preventing or treating a hypermelanosis disease, which includes administering a therapeutically effective amount of a pharmaceutical composition including a peptide consisting of the amino acid sequence of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3 as an effective ingredient to an individual.

[0057] Among the terms or technical features raised in the description regarding the pharmaceutical composition, the same terms or technical features as those already raised are as described above.

[0058] Another aspect provides a food composition for skin whitening, which includes a peptide consisting of the amino acid sequence of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3 as an effective ingredient.

[0059] Among the terms or technical features raised in the description regarding the peptide, the same terms or technical features as those already raised are as described above.

[0060] The content of the peptide included in the food composition is appropriately and non- limitingly selected according to the form of the food, the intended use, etc., and for example, can be added at 0.01 to 15% by weight based on the weight of the entire food. Also, the health beverage composition can be added at a ratio of 0.02 to 10 g, preferably 0.3 to 1 g per 100 ml.

[0061] Advantageous Effects

[0062] The peptide according to an aspect not only significantly inhibits melanin production and tyrosinase activity, but also inhibits the expression of factors related to melanin production, thereby showing excellent skin whitening effects.

[0063] The peptide according to an aspect not only significantly inhibits melanin production and tyrosinase activity, but also inhibits the expression of melanin production-related factors, and thus can be applied to alleviate or improve a hyperpigmentation disease of the skin.

[0064] Accordingly, the peptide according to an aspect can be used as an effective ingredient of a composition for skin whitening or a pharmaceutical composition for preventing or treating a hyperpigmentation disease. BRIEF DESCRIPTION OF DRAWINGS

[0065] Figure 1 Results of confirming a reduction in melanin production after adding a peptide consisting of the amino acid sequence of SEQ ID NO: 1 to B16F10 cells are shown.

[0066] Figure 2 Results of confirming a reduction in melanin production after adding a peptide consisting of the amino acid sequence of SEQ ID NO: 2 to B16F10 cells are shown.

[0067] Figure 3 Results of confirming a reduction in melanin production after adding a peptide consisting of the amino acid sequence of SEQ ID NO: 3 to B16F10 cells are shown.

[0068] Figure 4 Results of confirming a reduction in tyrosinase activity after adding a peptide consisting of the amino acid sequence of SEQ ID NO: 1 to B16F10 cells are shown.

[0069] Figure 5 Results of confirming a reduction in tyrosinase activity after adding a peptide consisting of the amino acid sequence of SEQ ID NO: 2 to B16F10 cells are shown.

[0070] Figure 6 Results of confirming a reduction in tyrosinase activity after adding a peptide consisting of the amino acid sequence of SEQ ID NO: 3 to B16F10 cells are shown.

[0071] Figure 7 Results of confirming the expression of melanin production-related genes, i.e., Rab27a, myosin VA, melanocyte-inducing transcription factor, tyrosinase-related protein 1, and tyrosinase, after adding a peptide consisting of the amino acid sequence of SEQ ID NO: 1 to B16F10 cells are shown.

[0072] Figure 8Results of confirming the inhibition of the expression of melanogenesis-related genes, i.e., melanin affinity, melanocyte-inducing transcription factor, and tyrosinase, after adding the peptide consisting of the amino acid sequence of SEQ ID NO: 2 to B16F10 cells are shown.

[0073] Figure 9 Results of confirming the inhibition of the expression of melanogenesis-related genes, i.e., Rab27a, myosin VA, melanin affinity, melanocyte-inducing transcription factor, tyrosinase, and blue aldehyde enzyme-related protein 1, after adding the peptide consisting of the amino acid sequence of SEQ ID NO: 3 to B16F10 cells are shown.

[0074] Figure 10 Results of confirming the inhibition of the expression of melanogenesis-related proteins, i.e., melanin affinity, tyrosinase, and melanocyte-inducing transcription factor, after adding the peptide consisting of the amino acid sequence of SEQ ID NO: 1 to B16F10 cells are shown.

[0075] Figure 11 Results of confirming the inhibition of the expression of melanogenesis-related proteins, i.e., tyrosinase, tyrosinase-related protein-1, and melanocyte-inducing transcription factor, after adding the peptide consisting of the amino acid sequence of SEQ ID NO: 2 to B16F10 cells are shown.

[0076] Figure 12 Results of confirming the inhibition of the expression of melanogenesis-related proteins, i.e., melanin affinity, Rab27a, tyrosinase, and melanocyte-inducing transcription factor, after adding the peptide consisting of the amino acid sequence of SEQ ID NO: 3 to B16F10 cells are shown. DETAILED DESCRIPTION

[0077] Hereinafter, the present application will be described in more detail by examples. However, these examples are only for illustratively describing the present application, and the scope of the present application is not limited to these examples.

[0078] Example 1, Synthesis of Peptide

[0079] The peptide having the amino acid sequence of SEQ ID NO: 1 recited in [Table 1] below was synthesized by using an automatic peptide synthesizer (Milligen 9050, Millipore, USA), and the synthesized peptide was purely isolated using a C18 reverse phase high performance liquid chromatography (High Performance Liquid Chromatography; HPLC) (Waters Associates, USA). ACQUITY UPLC BEH300 C18 (2.1 mm x 100 mm, 1.7 μm, Waters Co, USA) was used as a column.

[0080] [Table 1]

[0081] SEQ ID NO Sequence (N-terminal to C-terminal) 1 LRKRKRRFWVL 2 FHFIPNW 3 WPFLRP

[0082] Example 2, Confirmation of inhibitory effect on melanogenesis

[0083] B16F10 cells, which are mouse melanoma cells, were seeded at a density of 5 x 10 4 cells / well in a 6-well plate and cultured for 16 hours. Next, after replacing the culture medium with a culture medium including 2% serum, 200 ng / ml of α-melanocyte-stimulating hormones (α-MSH) was added to promote melanogenesis, and 10 μM, 50 μM, 100 μM, or 200 μM of a peptide composed of the amino acid sequence of SEQ ID NO: 1, 2, or 3 was added, respectively, and cultured for 72 hours. Next, the cultured cells were dissolved using 1 N NaOH, and the absorbance at 450 nm was measured. The untreated group (Con) was used as a control group, the group to which only α-melanocyte-stimulating hormones were added was used as a negative control group, and the group to which Arbutin (200 μM, 500 μM), which is known as a skin whitening agent, and α-melanocyte-stimulating hormones were added was used as a positive control group.

[0084] As a result, as shown in FIG. 1, the concentration of melanin was reduced in a concentration-dependent manner due to the addition of the peptide composed of the amino acid sequence of SEQ ID NO: 1, 2, or 3. Figures 1 to 3

[0085] Example 3, Confirmation of inhibitory effect on tyrosinase activity

[0086] B16F10 cells, which are mouse melanoma cells, were seeded at a density of 5 x 10 4 ​Cells were seeded at a density of 1 / well in 6-well plates and cultured for 16 hours. Then, the medium was replaced with a medium containing 2% serum, and 200 ng / ml of α-MSH was added, along with 1 μM, 10 μM, 20 μM, 40 μM, 60 μM, 80 μM, or 160 μM of a peptide consisting of amino acid sequences 1, 2, or 3, respectively, and the plates were cultured for 72 hours. The 6-well plates were placed on ice and washed with chilled PBS, and the cells were lysed by adding 0.1 M sodium phosphate buffer (pH 6.8, dissolution buffer) containing 1% Triton X-100. The cells present in the wells were then scraped and aggregated in a 1.5 ml tubing, vortexed, and centrifuged at 15,000 rpm for 10 minutes to obtain the supernatant. Next, the protein in the supernatant was quantified and the supernatant was adjusted to contain a certain amount of protein. Buffer was added, and a total of 90 μL of sample was aliquoted into a 96-well plate. Furthermore, the experimental sample, Blank sample, and positive control sample were prepared in the plate as shown in Table 2 below.

[0087] [Table 2]

[0088] Test sample Blank sample Positive control sample Sample 90 μl - - Buffer - 90 μl 90 μl Mushroom tyrosinase (0.1 mg / ml) - - 10 μl

[0089] Next, 20 μL of 10 mM L-DOPA was added to each sample, and after incubation at 37°C for one hour, the absorbance was measured at 475 nm. The untreated group (Con) was used as the control group, the group with only α-melanocyte-stimulating hormone (α-MSH) added was used as the negative control group, and the group with arbutin (200 μM or 500 μM) and α-MSH added was used as the positive control group.

[0090] As a result, it was confirmed that... Figures 4 to 6 As shown, the activity of tyrosinase is inhibited in a concentration-dependent manner by the addition of a peptide consisting of amino acid sequences of sequence numbers 1, 2, or 3.

[0091] Example 4: Confirmation of the inhibitory effect on the expression of melanin production-related genes.

[0092] Mouse melanoma cells, specifically B16F10 cells, were used at a rate of 5 × 10⁻⁶. 4Cells were seeded at a density of 1 / well in 6-well plates and cultured for 16 hours. Next, the medium was replaced with a medium containing 2% serum, and 200 ng / ml of α-melanocyte-stimulating hormone was added, along with 10 μM, 50 μM, 100 μM, or 200 μM of a peptide consisting of amino acid sequences 1, 2, or 3, respectively, and the plates were cultured for 72 hours. Messenger ribonucleic acid (RNA; mRNA) was extracted from the cultured cells, and the extracted mRNA was then reverse transcribed to synthesize complementary deoxyribonucleic acid (DNA; cDNA) using a cDNA synthesis kit & PCR pre-mix (Intron, Korea). Then, polymerase chain reaction (PCR) was performed using the complementary deoxyribonucleic acid and primers for Rab27a, melanophilin (Mlph), myosin VA, melanocyte-inducible transcription factor, tyrosinase, and tyrosinase-associated protein 1. The same groups used in Example 2 were used as the control, negative, and positive control groups, and the nucleotide sequences of the primers used in this example are shown in Table 3 below.

[0093] [Table 3]

[0094]

[0095]

[0096] The result is confirmed, such as Figure 7 As shown in Figure 8, the addition of a peptide consisting of the amino acid sequence of sequence number 1 inhibited the expression of melanin production-related genes, namely Rab27, myosin VA, melanocyte-inducible transcription factor, tyrosinase-related protein 1, and tyrosinase. Furthermore, as shown in Figure 8, the addition of a peptide consisting of the amino acid sequence of sequence number 2 inhibited the expression of melanin production-related genes, namely melanavidin, melanocyte-inducible transcription factor, and tyrosinase. Furthermore, as shown in Figure 8, the addition of a peptide consisting of the amino acid sequence of sequence number 2 inhibited the expression of melanin production-related genes, namely melanavidin, melanocyte-inducible transcription factor, and tyrosinase. Figure 9 As shown, the expression of melanin production-related genes, namely Rab27a, myosin VA, melanavitin, melanocyte-inducible transcription factor, tyrosinase, and tyrosinase-associated protein 1, was suppressed by adding a peptide consisting of the amino acid sequence of sequence number 3.

[0097] Example 5: Confirmation of the inhibitory effect on the expression of melanin production-related proteins.

[0098] Mouse melanoma cells, specifically B16F10 cells, were used at a rate of 5 × 10⁻⁶.4 Cells were seeded at a density of 100 / well in 6-well plates and cultured for 16 hours. Next, the medium was replaced with a medium containing 2% serum, to which 200 ng / ml of α-melanocyte-stimulating hormone was added, along with 10 μM, 50 μM, 100 μM, or 200 μM of a peptide consisting of amino acid sequences 1, 2, and 3, and the plates were cultured for 72 hours. Following lysis of the cultured cells, Western blotting was performed using antibodies against melanavirin, Rab27a, tyrosinase, melanocyte-inducing transcription factor, and tyrosinase-associated protein-1 (Santa Cruz Biotechnology, USA). The same control, negative control, and positive control groups used in Example 2 were employed.

[0099] The result is confirmed, such as Figure 10 As shown, the addition of a peptide consisting of the amino acid sequence of sequence number 1 inhibited the expression of melanocyte-related proteins melanocyte-binding protein, Rab27a, tyrosinase, and melanocyte-induced transcription factors. Furthermore, it was confirmed that, as... Figure 11 As shown, the addition of a peptide consisting of the amino acid sequence of sequence number 2 inhibited the expression of tyrosinase, tyrosinase-related protein-1, and melanocyte-induced transcription factors associated with melanin production. Furthermore, it was confirmed that... Figure 12 As shown, the expression of melanin-related proteins, namely melanavidin, Rab27a, tyrosinase, and melanocyte-induced transcription factors, was inhibited by adding a peptide composed of the amino acid sequence of sequence number 3.

[0100] Based on the experimental results, it can be concluded that the peptide with amino acid sequence number 1, 2, or 3 according to one embodiment has skin whitening effect.

[0101] The foregoing description of the present invention is merely exemplary, and those skilled in the art should understand that the invention can be modified into other specific forms without altering its technical concept or essential features. Therefore, the embodiments described above should be understood in all respects as exemplary, not restrictive.

Claims

1. A peptide consisting of the amino acid sequence shown in SEQ ID NO:

2.

2. A modified peptide formed by binding a protecting group selected from any one of acetyl, palmitoyl, myristyl, stearyl and polyethylene glycol to the N-terminal end of the peptide consisting of the amino acid sequence shown in SEQ ID NO:

2.

3. A modified peptide formed by binding an amino group to the C-terminal end of the peptide consisting of the amino acid sequence shown in SEQ ID NO:

2.

4. A composition for skin whitening, comprising: as an effective ingredient, the peptide according to claim 1 or the modified peptide according to claim 2 or 3, and a cosmetically acceptable carrier or a pharmaceutically acceptable carrier.

5. Use of the peptide consisting of the amino acid sequence shown in SEQ ID NO: 2 in the manufacture of a cosmetic composition for skin whitening.

6. Use of the peptide consisting of the amino acid sequence shown in SEQ ID NO: 2 in the manufacture of a pharmaceutical composition for the prophylaxis or treatment of a hypermelanosis disease.

7. The use according to claim 6, wherein the hypermelanosis disease is chloasma, freckle, senile lentigo or solar lentigo. ​ ​

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