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 invention patent application 201980038477.X, filed on October 10, 2019. Technical Field

[0002] This application relates to peptides with skin whitening activity and their uses. Background Technology

[0003] Besides differences in race, region, gender, and age, factors determining skin color generally include age spots, freckles, and tanning through exposure to ultraviolet light, as well as overall pigmentation, acne, scars, the distribution of keratinocytes, blood circulation, stress, and health status. Among these factors, pigmentation is known to be the primary factor in determining skin color.

[0004] Pigments that influence skin color include melanin, melanoidins, carotenoids, hemoglobin, and others. The various colors of skin, hair, and eyes depend on these pigments. Among them, melanin is the most important pigment in determining skin color; specifically, skin color depends on the amount and distribution of melanin. Melanin is produced in cells called melanocytes under the epidermis and is moved to the stratum corneum and shed due to skin metabolism. Regardless of skin color, the number of melanocytes is almost the same; differences in skin color arise solely from variations in the production, type, and distribution of melanin.

[0005] In the skin, tyrosine is converted into dihydroxyphenylalanine (DOPA) by an enzyme called tyrosinase, and through a series of oxidation processes, it ultimately produces melanin, a dark brown polymer. Excessive melanin production can lead to hyperpigmentation, resulting in age spots, freckles, and other blemishes, causing undesirable cosmetic outcomes. Currently, with the increase in the leisure population and the growing number of people enjoying outdoor activities, the demand for preventing melanin deposition caused by ultraviolet radiation is also increasing.

[0006] Against this technological backdrop, research is underway from multiple angles to develop whitening agents that prevent excessive melanin production through mechanisms such as inhibiting the activity of melanocyte-producing cells and inhibiting tyrosinase activity (Korean Patent Publication No. 10-2019-0109722), but it is still insignificant. Summary of the Invention

[0007] Technical issues

[0008] One aspect provides a peptide composed of an amino acid sequence of sequence number 1, sequence number 2, or sequence number 3.

[0009] On the other hand, a composition for skin whitening is provided, comprising a peptide composed of an amino acid sequence consisting of serial numbers 1, 2, and 3 as an active ingredient.

[0010] On the other hand, a cosmetic composition is provided that includes the aforementioned skin-whitening composition.

[0011] On the other hand, a pharmaceutical composition for the prevention or treatment of melanin hyperpigmentation is provided, comprising the composition for skin whitening.

[0012] Other objects and advantages of this application will become more apparent from the appended claims and drawings, as well as the following detailed description. For matters not described herein, which will be readily apparent and analogous to those skilled in the art to which this application pertains or similar fields, description is omitted.

[0013] Technical solution

[0014] The descriptions and embodiments disclosed in this application can be applied to other descriptions and embodiments. In other words, all combinations of the various technical features disclosed in this application fall within the scope of this application. Furthermore, the scope of this application is not limited to the specific descriptions set forth below.

[0015] On the one hand, it provides peptides composed of amino acid sequences of sequence number 1, sequence number 2, or sequence number 3.

[0016] As used in this specification, the term "peptide" can refer to a linear molecule formed by the binding of amino acid residues together via peptide bonds. The peptide can be prepared according to chemical synthesis methods 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, Second Edition, Pierce Chem. Co.: Rockford, 111 (1984)) or liquid-phase synthesis techniques (US Patent No. 5,516,891). The inventors have devoted considerable effort to developing a peptide with biologically effective activity, and as a result, have identified a peptide composed of an amino acid sequence of Serial No. 1, Serial No. 2, or Serial No. 3. Here, the biologically effective activity can refer to exhibiting any one or more of the following properties: (a) inhibition of melanin production; (b) inhibition of tyrosinase activity; and (c) inhibition of the expression of Rab27a, melanavirin, myosin VA, melanocyte-inducible transcription factor, tyrosinase, or tyrosinase-associated protein 1. Therefore, the peptide is used for skin whitening, prevention, or treatment of melanin hyperpigmentation disorders.

[0017] To achieve chemical stability, enhanced pharmacological properties (half-life, absorption, titer, potency, etc.), altered specificity (e.g., broad bioactivity profile), and reduced antigenicity, a protecting group may be bound to the N- or C-terminus of the peptide. In one specific embodiment, the N-terminus of the peptide is bound to any one of the following protecting groups: acetyl group, fluorenylmethoxycarbonyl group, formyl group, palmitoyl group, myristyl group, stearyl group, butoxycarbonyl group, allyloxycarbonyl group, and polyethylene glycol (PEG); and / or the C-terminus of the peptide is bound to any one of the following protecting groups: amino group (-NH2), tertiary alkyl group, and azide (-NHNH2). Furthermore, the peptide may optionally include a guide sequence, a tag, labeled residues, a half-life, or an amino acid sequence manufactured for a specific purpose of improving peptide stability.

[0018] The term "stability" as used in this specification can refer not only to the in vivo stability of the peptide against attack by proteases in vivo, but also to storage stability (e.g., room temperature storage stability).

[0019] Other aspects include: a cosmetic composition comprising a peptide comprising an amino acid sequence including sequence number 1, sequence number 2, or sequence number 3 as an active ingredient; a pharmaceutical composition comprising a peptide comprising an amino acid sequence including sequence number 1, sequence number 2, or sequence number 3 as an active ingredient; and use of a peptide comprising an amino acid sequence including sequence number 1, sequence number 2, or sequence number 3 for use in the preparation of a cosmetic or pharmaceutical composition or as a cosmetic or pharmaceutical composition.

[0020] On the other hand, a composition for skin whitening is provided, comprising a peptide consisting of an amino acid sequence of sequence number 1, sequence number 2, or sequence number 3 as an active ingredient.

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

[0022] The term "skin whitening" as used in this specification can be interpreted as not only inhibiting melanin synthesis to brighten skin tone but also improving hyperpigmentation of the skin due to ultraviolet radiation, hormones, or genetics. Hyperpigmentation of the skin may include, but is not limited to, freckles, melasma, age spots, brown spots, or freckles.

[0023] The term “improvement” as used in this specification may refer to all behaviors that alleviate or at least reduce the severity of treatment-related parameters such as symptoms.

[0024] While existing functional peptides possess effective biological activity, their size limits their effective delivery to target tissues or cells, or their short half-life leads to rapid elimination in the body. In contrast, a skin-whitening composition according to one embodiment comprises a peptide consisting of 10 or fewer amino acids as the active ingredient. Therefore, the skin penetration rate of the active ingredient is excellent, and for example, when applied topically to the skin, an effective skin-whitening effect can be achieved.

[0025] According to one embodiment, the peptide can significantly inhibit melanin production and tyrosinase activity. Furthermore, the peptide can inhibit the expression of melanin-related factors, namely 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 active ingredient in cosmetic compositions for skin whitening.

[0026] On the other hand, a cosmetic composition for skin whitening is provided, comprising a peptide consisting of an amino acid sequence of serial number 1, serial number 2, or serial number 3 as an active ingredient.

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

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

[0029] The term "cosmetically effective amount" as used in this specification refers to an amount sufficient to achieve the skin-whitening effect of the cosmetic composition.

[0030] The weight ratio of the peptide to the cosmetically 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.

[0031] The cosmetic composition can be formulated into any dosage form commonly prepared in the art, such as, but not limited to, solutions, suspensions, emulsions, doughs, gels, creams, lotions, powders, soaps, detergents including surfactants, oils, powder-type foundations, emulsion-type foundations, wax-type foundations, and hairsprays. For example, the cosmetic composition can be formulated into softening lotions, nourishing lotions, nourishing creams, massage creams, serums, eye creams, cleansing pastes, facial cleansers, cleansing waters, masks, hairsprays, or foundations.

[0032] When the dosage form of the cosmetic composition is dough, cream, or gel, animal oil, vegetable oil, beeswax, paraffin, starch, tragacanth gum, cellulose derivatives, polyethylene glycol, bentonite, silica, talc, or zinc oxide may be used as carrier ingredients.

[0033] When the cosmetic composition is in the form of a powder or hairspray, lactose, talc, silica, aluminum hydroxide, calcium silicate, or polyamine powder may be used as a carrier component. For example, when the cosmetic composition is a hairspray, it may further include a propellant such as chlorofluorocarbon, propane / butane, or dimethyl ether.

[0034] When the dosage form of the cosmetic composition is a solution or emulsion, a solvent, dissolving agent, or emulsifier may be used as a carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butanediol oil, glycerol aliphatic ester, polyethylene glycol, or fatty acid esters of sorbitan.

[0035] When the dosage form of the cosmetic composition is a suspension, liquid phase diluents such as water, ethanol or propylene glycol, suspending agents such as ethoxylated isooctadecyl alcohol, polyoxyethylene sorbitan ester, microcrystalline cellulose, aluminum hydroxide, bentonite, agar or tragacanth gum, etc., can be used as carrier components.

[0036] When the dosage form of the cosmetic composition is a detergent containing surfactants, aliphatic alcohol sulfates, aliphatic alcohol ether sulfates, monosuccinate, ethanesulfonate, imidazoline derivatives, methyl taurate, sarcosinate, aliphatic amide ether sulfates, cocoa polysaccharides, fatty alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives, or ethoxylated propylene glycol fatty acid esters may be used as carrier components.

[0037] In addition to peptides and carrier components that are active ingredients, the ingredients included in the cosmetics include ingredients commonly used in cosmetic compositions, such as conventional adjuvants like antioxidants, stabilizers, solvents, vitamins, pigments, and fragrances.

[0038] On the other hand, a skin whitening method is provided, which includes applying a cosmetic composition comprising a peptide consisting of an amino acid sequence of sequence number 1, sequence number 2, or sequence number 3 as an active ingredient to an individual's skin.

[0039] In the description of the cosmetic composition, the terms or technical features that are the same as those already mentioned are as described above.

[0040] The terms “applied,” “applied,” and “coated” used in this specification are used interchangeably and may refer to localization of at least a portion of a desired site by inducing the application of a composition according to an embodiment to a particular site, or may refer to the configuration of a composition according to an embodiment into an individual according to an application route.

[0041] On the other hand, a pharmaceutical composition for the prevention or treatment of melanin hyperpigmentation is provided, comprising a peptide consisting of an amino acid sequence of sequence number 1, sequence number 2, or sequence number 3 as an active ingredient.

[0042] In the description of the peptide, the same terms or technical features as those already mentioned are as described above.

[0043] In this specification, the term "prevention" refers to all actions that suppress or delay the occurrence of disease by applying the composition.

[0044] In this specification, the term "treatment" refers to any form of treatment that provides an individual with a disease or at risk of developing a disease with effects including improvement of the individual's condition (e.g., more than one symptom), delay of disease progression, delay of symptom onset, or attenuation of symptom progression. Therefore, "treatment" and "prevention" do not refer to a cure or complete removal of symptoms.

[0045] The term "individual" refers to an object requiring treatment for a disease, more specifically to humans or non-human primates, mice, dogs, hairy animals, horses, and other mammals such as cattle.

[0046] As a disease that needs to be prevented or treated by the pharmaceutical composition, "melanosis disease" can be a general term for diseases in which melanin is produced and deposited excessively on the skin, such as melasma, freckles, senile lentigines, or solar lentigines, but is not limited thereto.

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

[0048] The term "effective amount of drug" as used in this specification may refer to an amount sufficient to achieve the preventive or therapeutic efficacy of the pharmaceutical composition against melanin hyperpigmentation.

[0049] The weight ratio of the peptide to 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] Pharmaceutically acceptable carriers are those commonly used in the preparation of dosage forms, including, but not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, starch, gum arabic, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methylcellulose, methylparaben, propylparaben, talc, magnesium stearate, and mineral oil. Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington's Pharmaceutical Sciences (19th edition, 1995).

[0051] In addition to the components mentioned above, the pharmaceutical composition may further include, but is not limited to, lubricants, humectants, sweeteners, flavorings, emulsifiers, suspending agents, preservatives, etc.

[0052] The pharmaceutical composition can be administered orally or parenterally, preferably parenterally. When administered parenterally, it can be administered by injection into an intramuscular, intravenous, subcutaneous, or peritoneal cavity, local application, or percutaneous application, but is not limited thereto.

[0053] The dosage of the pharmaceutical composition may be 0.0001 to 1000 μg, 0.001 to 1000 μg, 0.01 to 1000 μg, 0.1 to 1000 μg, or 1.0 to 1000 μg per day, but is not limited thereto, and may be prescribed differently depending on factors such as formulation method, administration method, patient age, weight, gender, symptoms, diet, administration time, administration route, excretion rate, and reaction sensitivity.

[0054] The pharmaceutical composition is prepared by means of a method readily practiced by a person skilled in the art to which this invention pertains, by formulation using pharmaceutically acceptable carriers and / or excipients to produce a unit volume form or to be injected into a large volume container.

[0055] The dosage form may be a solution, suspension or emulsion in an oil or aqueous medium, or it may be an extract, powder, granule, tablet or capsule, and may further include dispersants and / or stabilizers.

[0056] On the other hand, methods for preventing or treating melanin hyperpigmentation include administering a therapeutically effective amount of a pharmaceutical composition to an individual, comprising a peptide consisting of an amino acid sequence of sequence number 1, sequence number 2, or sequence number 3 as an active ingredient.

[0057] In the description of the pharmaceutical composition, the same terms or technical features as those already mentioned are as described above.

[0058] On the other hand, a food composition for skin whitening is provided, comprising a peptide composed of an amino acid sequence of serial number 1, serial number 2, or serial number 3 as an active ingredient.

[0059] In the description of the peptide, the same terms or technical features as those already mentioned are as described above.

[0060] The amount of peptides included in the food composition can be appropriately and non-limitingly selected according to the form of the food, the intended use, etc., for example, it can be added at a rate of 0.01 to 15% by weight of the total food. Furthermore, based on 100 ml, it can be added to the health beverage composition at a rate of 0.02 to 10 g, preferably 0.3 to 1 g.

[0061] Beneficial effects

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

[0063] Based on the fact that one type of peptide not only significantly inhibits melanin production and tyrosinase activity, but also inhibits the expression of melanin production-related factors, it can be applied to alleviate or improve melanin hyperpigmentation disorders of the skin.

[0064] Therefore, peptides can be used as active ingredients in compositions for skin whitening or in pharmaceutical compositions for the prevention or treatment of hypermelanosis. Attached Figure Description

[0065] Figure 1 The results show a reduction in melanin production after adding a peptide consisting of the amino acid sequence of sequence number 1 to B16F10 cells.

[0066] Figure 2 The results show a reduction in melanin production after adding a peptide consisting of the amino acid sequence of sequence number 2 to B16F10 cells.

[0067] Figure 3 The results show a reduction in melanin production after adding a peptide consisting of the amino acid sequence of sequence number 3 to B16F10 cells.

[0068] Figure 4 The results show a reduction in tyrosinase activity after adding a peptide consisting of the amino acid sequence of sequence number 1 to B16F10 cells.

[0069] Figure 5 The results show a reduction in tyrosinase activity after adding a peptide consisting of the amino acid sequence of sequence number 2 to B16F10 cells.

[0070] Figure 6 The results show a reduction in tyrosinase activity after adding a peptide consisting of the amino acid sequence of sequence number 3 to B16F10 cells.

[0071] Figure 7 The results show the expression of melanin production-related genes, namely Rab27a, myosin VA, melanocyte-inducible transcription factor, tyrosinase-associated protein 1, and tyrosinase, after adding the peptide consisting of the amino acid sequence of sequence number 1 to B16F10 cells.

[0072] Figure 8The results show that adding the peptide consisting of the amino acid sequence of sequence number 2 to B16F10 cells confirmed the inhibition of the expression of melanin-related genes, namely melanavidin, melanocyte-inducible transcription factor, and tyrosinase.

[0073] Figure 9 The results show the inhibition of the expression of melanin-related genes Rab27a, myosin VA, melanavitin, melanocyte-inducible transcription factor, tyrosinase, and tyrosinase-associated protein 1 after adding the peptide consisting of the amino acid sequence of sequence number 3 to B16F10 cells.

[0074] Figure 10 The results show that adding the peptide consisting of the amino acid sequence of sequence number 1 to B16F10 cells confirmed the inhibition of the expression of melanin-related proteins, namely melanavidin, tyrosinase, and melanocyte-induced transcription factors.

[0075] Figure 11 The results show the inhibition of the expression of melanin-related proteins, namely tyrosinase, tyrosinase-associated protein-1, and melanocyte-induced transcription factors, after adding the peptide consisting of the amino acid sequence of sequence number 2 to B16F10 cells.

[0076] Figure 12 The results show the inhibition of the expression of melanin-related proteins, namely melanavidin, Rab27a, tyrosinase, and melanocyte-induced transcription factors, after adding the peptide consisting of the amino acid sequence of sequence number 3 to B16F10 cells. Detailed Implementation

[0077] The invention is described in more detail below by way of examples. However, these examples are for illustrative purposes only, and the scope of the invention is not limited to these examples.

[0078] Example 1: Peptide Synthesis

[0079] Peptides with the amino acid sequences listed in Table 1 below, containing sequence number 1, were synthesized using an automated peptide synthesizer (Milligen 9050, Millipore, USA) and purified using C18 reversed-phase high-performance liquid chromatography (HPLC) (Waters Associates, USA). An ACQUITY UPLC BEH300 C18 column (2.1 mm × 100 mm, 1.7 μm, Waters Co., USA) was used.

[0080] [Table 1]

[0081] Serial Number Sequence (N-end → C-end) 1 LRKRKRRFWVL 2 FHFIPNW 3 WPFLRP

[0082] Example 2: Confirmation of the inhibitory effect on melanin production

[0083] Mouse melanoma cells, specifically B16F10 cells, were used at a rate of 5 × 10⁻⁶. 4 Cells were seeded at a density of 2% per well in 6-well plates and cultured for 16 hours. Then, after replacing the medium with a medium containing 2% serum, 200 ng / ml of α-melanocyte-stimulating hormones (α-MSH) was added to promote melanin production. Simultaneously, 10 μM, 50 μM, 100 μM, or 200 μM of a peptide consisting of amino acid sequences 1, 2, or 3 were added, and the plates were cultured for 72 hours. The cultured cells were then dissolved in 1N NaOH, and absorbance was measured at 450 nm. An untreated group (Con) served as the control, a group receiving only α-MSH served as the negative control, and groups receiving arbutin (200 μM, 500 μM), a known skin-brightening agent, and α-MSH served as the positive control.

[0084] As a result, it was confirmed that... Figures 1 to 3 As shown, the concentration of melanin production is reduced in a concentration-dependent manner by the addition of a peptide consisting of amino acid sequences of sequence numbers 1, 2, or 3.

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

[0086] 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. 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 samples Blank sample Positive control group samples Sample 90μl - - buffer solution - 90μl 90μl Mushroom tyrosinase (0.1 mg / ml) - - 10μl

[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 Serial No.

3.

2. A modified peptide, which is formed by binding a peptide consisting of the amino acid sequence shown in Serial No. 3 to an N-terminus with a protecting group selected from any of the following: acetyl, palmitoyl, myristyl, stearyl, and polyethylene glycol.

3. A modified peptide, which is formed by the amino acid sequence shown in Serial No. 3 being bound to an amino group at the C-terminus.

4. A composition for skin whitening, comprising: The peptide according to claim 1 or the modified peptide according to claim 2 or 3 as the active ingredient, and A carrier that is acceptable in cosmetics or pharmaceuticals.

5. Use of the peptide consisting of the amino acid sequence shown in Serial No. 3 in the preparation of cosmetic compositions for skin whitening.

6. Use of a peptide consisting of the amino acid sequence shown in Serial No. 3 in the preparation of a pharmaceutical composition for the prevention or treatment of melanin hyperpigmentation.

7. The use according to claim 6, wherein the melanin hyperpigmentation disease is melasma, freckles, senile lentigines, or solar lentigines.

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