Peptide having skin-whitening activity and uses thereof

Abstract

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

Description

Peptides having skin whitening activity and their uses

[0001] The present application relates to a peptide having skin whitening activity and the use thereof.

[0002] Factors determining skin color, aside from differences based on race, region, gender, and age, primarily include melasma, freckles, and tanning caused by UV exposure. Other factors include overall pigmentation, acne, scars, keratin distribution, blood circulation, stress, and health status. Among these factors, pigmentation is known to be a major determinant of skin color.

[0003] Pigments that influence skin color include melanin, melanoids, carotene, hemoglobin, and carotenoids, and the various colors of the skin, hair, and eyes are determined by these pigments. Among these, melanin is the most important pigment determining skin color; specifically, skin color is determined by the amount and distribution of melanin. Melanin is produced by cells called melanocytes located beneath the epidermis and migrates to the surface of the stratum corneum through skin metabolism before sloughing off. Regardless of skin color, the number of melanocytes remains nearly the same; however, skin color varies simply because the amount, type, and distribution of melanin produced differ.

[0004] In the skin, tyrosine is converted into DOPA by a human enzyme called tyrosinase, and through a series of subsequent oxidation processes, melanin, a dark brown polymer, is finally produced. When melanin is produced in excess of what is necessary, it causes hyperpigmentation such as melasma, freckles, and moles, leading to undesirable cosmetic results. Meanwhile, with the recent increase in the leisure population and the growing number of people enjoying outdoor activities, there is a rising demand to prevent melanin pigmentation caused by ultraviolet rays.

[0005] Against this technological backdrop, multifaceted research is being conducted on the development of whitening agents that prevent excessive melanin production (Korean Published Patent No. 10-2019-0050058), but the current situation is still insufficient.

[0006] One aspect is to provide a peptide composed of the amino acid sequence of SEQ ID NO. 1.

[0007] Another aspect is to provide a skin whitening composition comprising a peptide consisting of the amino acid sequence of SEQ ID NO. 1 as an active ingredient.

[0008] Another aspect is to provide a cosmetic composition comprising the above-mentioned skin whitening composition.

[0009] Another aspect is to provide a pharmaceutical composition for the prevention or treatment of melanin hyperpigmentation disorders comprising the above-mentioned skin whitening composition.

[0010] Another aspect provides a skin whitening method comprising the step of applying a cosmetic composition containing a peptide having the amino acid sequence of SEQ ID NO. 1 as an active ingredient to the skin of an individual.

[0011] Another aspect provides a method for preventing or treating hypermelanosis, comprising the step of administering to an individual a pharmaceutical composition containing a peptide having the amino acid sequence of SEQ ID NO. 1 as an active ingredient.

[0012] Another aspect is to provide a use of a peptide consisting of the amino acid sequence of SEQ ID NO. 1 for the preparation of a cosmetic composition for skin whitening or a pharmaceutical composition for the prevention or treatment of hypermelanin pigmentation disorders.

[0013] Other objects and advantages of this application will become more apparent from the following detailed description, together with the appended claims and drawings. Anything not described in this specification is omitted, as it can be sufficiently recognized and inferred by those skilled in the art of this application or a similar art field.

[0014] Each description and embodiment disclosed in this application may be applied to each other description and embodiment. That is, all combinations of the various elements disclosed in this application fall within the scope of this application. Furthermore, the scope of this application should not be considered limited by the specific descriptions provided below.

[0015] One aspect provides a peptide composed of the amino acid sequence of SEQ ID NO. 1.

[0016] As used herein, the term "peptide" may mean a linear molecule formed by amino acid residues being joined together by peptide bonds. The peptide may be prepared according to chemical synthesis methods known in the art, in particular 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, 111(1984)) or liquid-phase synthesis techniques (US Patent No. 5,516,891).

[0017] As a result of diligent efforts to develop a peptide having biologically effective activity, the inventors identified a peptide composed of the amino acid sequence of SEQ ID NO. 1. Herein, the biologically effective activity may exhibit one or more selected from characteristics such as (a) inhibition of melanin production; (b) inhibition of CREB (cAMP response element-binding protein) phosphorylation; (c) inhibition of MITF (Microphthalmia-associated transcription factor), TYR (tyrosinase), or TYRP1 (tyrosinase related protein 1) expression; and (d) inhibition of Rab27a, MYO5A (myosin 5a), or MLPH (Melanophilin) ​​expression. Accordingly, the peptide may be utilized for skin whitening, or for the prevention or treatment of melanin hyperpigmentation disorders.

[0018] The above peptide may have a protecting group attached to the N- or C-terminus of the peptide to obtain chemical stability, enhanced pharmacological properties (half-life, absorption, potency, efficacy, etc.), modified specificity (e.g., broad spectrum of biological activity), and reduced antigenicity. The protecting group may be a protecting group that is not an amino acid. In one embodiment, the N-terminus of the peptide may be attached 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 polyethylene glycol (PEG); and / or the C-terminus of the peptide may be bonded to any one of the protecting groups selected from the group consisting of an amino group (-NH2), a tertiary alkyl group, and a hydrazino group (-NHNH2). Additionally, the peptide may optionally further include a targeting sequence, a tag, a labeled residue, an amino acid sequence prepared for a specific purpose to increase half-life or peptide stability.

[0019] The above peptide is artificially synthesized, or non-naturally occurring or engineered, and the term “non-naturally occurring or engineered” means a state created by applying artificial modifications rather than the state in which it exists naturally. Here, the artificial modification may include artificially synthesizing an amino acid sequence by mimicking the structure of multiple amino acids, or being engineered to obtain chemical stability, enhanced pharmacological properties, altered specificity, or reduced antigenicity as described above.

[0020] The term "stability" as used in this specification may mean not only in vivo stability, which protects the peptide from attack by protein-cleaving enzymes in vivo, but also storage stability (e.g., room temperature storage stability).

[0021] Another aspect provides a skin whitening composition comprising a peptide consisting of the amino acid sequence of SEQ ID NO. 1 as an active ingredient.

[0022] Among the terms or elements mentioned in the description of the above peptide, those identical to those already mentioned are as previously stated.

[0023] As used herein, the term "skin whitening" may be interpreted as not only brightening the skin tone by inhibiting the synthesis of melanin pigment, but also improving hyperpigmentation of the skin caused by ultraviolet rays, hormones, or heredity. Here, the hyperpigmentation of the skin may include, but is not limited to, freckles, melasma, senile lentigo, brown spots, or age spots.

[0024] As used in this specification, the term "improvement" may mean any action that at least reduces parameters related to the alleviation or treatment of a condition, such as the degree of symptoms.

[0025] Conventional functional peptides, despite their effective biological activity, have the disadvantage of not being effectively absorbed into target tissues or cells due to the size of the peptide itself, or being eliminated from the body in a short period of time due to their short half-life. On the other hand, a whitening composition according to one embodiment includes a peptide composed of 10 or fewer amino acids as an active ingredient, and accordingly, 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 obtained.

[0026] According to one embodiment, the peptide can inhibit melanin-producing factors including MITF, TYR, or TYRP1, and inhibit melanosome migration-related inhibitory factors including Rab27a, MYO5A, or MLPH. The peptide can exhibit skin whitening efficacy through the inhibition of melanin production and melanosome migration. Therefore, the peptide can be utilized as an active ingredient in a skin whitening composition.

[0027] Another aspect provides a cosmetic composition for skin whitening comprising a peptide consisting of the amino acid sequence of SEQ ID NO. 1 as an active ingredient.

[0028] Among the terms or elements mentioned in the description of the above peptide, those identical to those already mentioned are as previously stated.

[0029] The above cosmetic composition may comprise a cosmetically effective amount of the peptide; and / or a cosmetically acceptable carrier, but is not limited thereto.

[0030] The above carrier may promote local delivery of the above peptide.

[0031] As used in this specification, the term "cosmetic effective amount" means an amount sufficient to achieve skin whitening efficacy of the cosmetic composition.

[0032] The weight ratio between the above peptide and the cosmetically acceptable carrier may be, for example, 500:1 to 1:500, and as an example, the weight ratio may 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 It may be 2:1 to 1:2, but is not limited thereto.

[0033] The above cosmetic composition may be prepared in any formulation conventionally manufactured in the art, for example, as a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing, oil, powder foundation, emulsion foundation, wax foundation, and spray, but is not limited thereto. For example, it may be prepared in the form of a softening lotion, a nourishing lotion, a nourishing cream, a massage cream, an essence, an eye cream, a cleansing cream, a cleansing foam, a cleansing water, a pack, a spray, or a powder.

[0034] In the case where the formulation of the above cosmetic composition is a paste, cream, or gel, animal oil, vegetable oil, wax, paraffin, starch, tracanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, or zinc oxide may be used as a carrier component.

[0035] When the formulation of the above cosmetic composition is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, or polyamide powder may be used as a carrier component, and, for example, in the case of a spray, it may additionally include a propellant such as chlorofluorohydrocarbon, propane / butane, or dimethyl ether.

[0036] When the formulation of the above cosmetic composition is a solution or an emulsion, a solvent, a solubilizing agent, or an emulsifying agent is used as a carrier component, and may include, for example, water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol, or fatty acid ester of sorbitan.

[0037] In the case where the formulation of the above cosmetic composition is a suspension, liquid diluents such as water, ethanol, or propylene glycol, suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, and polyoxyethylene sorbitan ester, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar, or tracanth may be used as carrier components.

[0038] In the case where the formulation of the above cosmetic composition is a cleansing product containing a surfactant, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyl taurate, sarcosinate, fatty acid amide ether sulfate, alkylamidobetaine, aliphatic alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, lanolin derivative, or ethoxylated glycerol fatty acid ester, etc. may be used as a carrier component.

[0039] The above peptide may be contained in nanosomes or nanoparticles to further improve skin penetration or stability issues. For example, the nanosomes may be manufactured using a microfluidizer with lecithin as a raw material and may be contained within lecithin particles. Any known method may be used to manufacture the nanosomes. The size of the nanosome particles is preferably 30 to 200 nm. If the size of the nanosome particles is less than 30 nm, skin penetration proceeds very quickly, which may cause skin side effects, and if it exceeds 200 nm, skin penetration is not easy, making it difficult to obtain the effect of using the nanosome structure.

[0040] In addition to the peptide and carrier components as active ingredients, the components included in the above cosmetic composition include ingredients commonly used in cosmetic compositions, and may include, for example, conventional auxiliary agents such as antioxidants, stabilizers, solubilizers, vitamins, pigments, and fragrances.

[0041] The content of the peptide as an active ingredient contained in the above cosmetic composition can be appropriately and unrestrictively selected depending on the form of the product, the desired use, etc., and, for example, can be added in an amount of 0.01 to 15 weight% of the total weight of the cosmetic composition. In addition, for example, the cosmetic composition may contain peptide in an amount of 1.0 weight% to 3.0 weight%, preferably 2.0 weight% to 3.0 weight%, based on the total weight, but is not limited thereto.

[0042] Another aspect provides a skin whitening method comprising the step of applying a cosmetic composition containing a peptide having the amino acid sequence of SEQ ID NO. 1 as an active ingredient to the skin of an individual.

[0043] Among the terms or elements mentioned in the description of the above cosmetic composition, those identical to those already mentioned are as previously stated.

[0044] The terms “applying,” “administering,” and “applying” as used herein are used interchangeably and may mean causing at least partial localization of the composition according to one embodiment to a desired site, or dispensing the composition according to one embodiment into an individual by a route of administration.

[0045] Another aspect provides a pharmaceutical composition for preventing or treating hypermelanosis of pigmentation disease comprising a peptide consisting of the amino acid sequence of SEQ ID NO. 1 as an active ingredient.

[0046] Among the terms or elements mentioned in the description of the above peptide, those identical to those already mentioned are as previously stated.

[0047] In this specification, the term "prevention" refers to any act of suppressing or delaying the onset of a disease through the administration of the above composition.

[0048] In this specification, the term “treatment” means any form of treatment that provides effects to an individual suffering from a disease or at risk of developing a disease, including improvement of the individual’s condition (e.g., one or more symptoms), delay of disease progression, delay of symptom onset, or slowing of symptom progression. Accordingly, “treatment” and “prevention” are not intended to mean the cure or complete elimination of symptoms.

[0049] The above "individual" refers to a subject requiring treatment for a disease, and more specifically, to mammals such as human or non-human primates, mice, dogs, cats, horses, and cattle.

[0050] The "melanin hyperpigmentation disease," which is a disease to be prevented or treated by the above pharmaceutical composition, may be a general term for a disease in which melanin pigment is excessively produced and deposited in the skin, and may be, for example, melasma, freckles, senile pigment spots, or solar lentigines, but is not limited thereto.

[0051] The above pharmaceutical composition may comprise a pharmaceutically effective amount of the peptide; and / or a pharmaceutically acceptable carrier, but is not limited thereto.

[0052] The above carrier may promote local delivery of the above peptide.

[0053] As used in this specification, the term "pharmaceutical effective amount" may mean an amount sufficient to achieve the efficacy of preventing or treating melanin hyperpigmentation disease of the pharmaceutical composition.

[0054] The above pharmaceutically acceptable carriers are those commonly used in formulations and include, but are not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methylcellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil. Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington's Pharmaceutical Sciences (19th ed., 1995).

[0055] The weight ratio between the above peptide and the pharmaceutically acceptable carrier may be, for example, 500:1 to 1:500, and as an example, said weight ratio may 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 It may be 2:1 to 1:2, but is not limited thereto.

[0056] The above pharmaceutical composition may additionally include, but is not limited to, lubricants, wetting agents, sweeteners, flavoring agents, emulsifiers, suspending agents, preservatives, etc. in addition to the above components.

[0057] The above pharmaceutical composition may be administered orally or parenterally, preferably parenterally. In the case of parenteral administration, it may be administered by intramuscular injection, intravenous injection, subcutaneous injection, intraperitoneal injection, local administration, transdermal administration, etc., but is not limited thereto.

[0058] The dosage of the above pharmaceutical composition may be 0.0001 to 1000 µg (micrograms, 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 in various ways depending on factors such as the formulation method, method of administration, age, weight, sex, pathological condition of the patient, food, time of administration, route of administration, excretion rate and response responsiveness.

[0059] The above pharmaceutical composition may be manufactured in a unit volume form or contained in a multi-volume container by formulation using a pharmaceutically acceptable carrier and / or excipient, according to a method that can be easily carried out by a person skilled in the art to which the invention belongs.

[0060] The above formulation may be in the form of a solution, suspension, or emulsion in an oil or aqueous medium, or may be in the form of an extract, powder, granule, tablet, or capsule, and may additionally include a dispersant and / or stabilizer.

[0061] Another aspect provides a method for preventing or treating hypermelanosis, comprising the step of administering to an individual a pharmaceutical composition containing, as an active ingredient, a therapeutically effective amount of a peptide consisting of the amino acid sequence of SEQ ID NO. 1.

[0062] Among the terms or elements mentioned in the description of the above pharmaceutical composition, those identical to those already mentioned are as previously stated.

[0063] Another aspect is to provide a use of a peptide consisting of the amino acid sequence of SEQ ID NO. 1 for the preparation of a cosmetic composition for skin whitening or a pharmaceutical composition for the prevention or treatment of hypermelanin pigmentation disorders.

[0064] Among the terms or elements mentioned in the description of the above use, those identical to those already mentioned are as previously stated.

[0065] Another aspect provides a food composition for skin whitening comprising a peptide consisting of the amino acid sequence of SEQ ID NO. 1 as an active ingredient.

[0066] Among the terms or elements mentioned in the description of the above peptide, those identical to those already mentioned are as previously stated.

[0067] The content of the peptide as an active ingredient contained in the above food composition can be appropriately and unrestrictively selected depending on the form of the food, the desired use, etc., and, for example, can be added in an amount of 0.01 to 15 weight% of the total food weight. In addition, for example, the health drink composition can be added in a ratio of 0.02 to 10 g, preferably 0.3 to 1 g, based on 100 ml, but is not limited thereto.

[0068] According to a peptide according to one aspect, by not only inhibiting the expression of factors related to melanin production but also inhibiting factors related to melanosome migration, it exhibits an excellent skin whitening effect and can be applied to alleviate or improve skin hypermelanin pigmentation disorders.

[0069] Accordingly, a peptide according to one aspect can be utilized as an active ingredient in a skin whitening composition or a pharmaceutical composition for the prevention or treatment of melanin hyperpigmentation disorders.

[0070] Figure 1 is the result of confirming the reduction in melanin production after adding a peptide consisting of the amino acid sequence of SEQ ID NO. 1 to B16F10 cells.

[0071] Figure 2 shows the results of confirming the phosphorylation level of CREB by Western blot after adding a peptide consisting of the amino acid sequence of SEQ ID NO. 1 to B16F10 cells.

[0072] Figure 3 shows the results of confirming the expression levels of MITF, TYR, and TYRP1, which are melanin-producing genes, through RT-PCR after adding a peptide consisting of the amino acid sequence of SEQ ID NO. 1 to B16F10 cells.

[0073] Figure 4 shows the results of confirming the expression levels of MITF, TYR, and TYRP1, which are melanin-producing proteins, by Western blot after adding a peptide consisting of the amino acid sequence of SEQ ID NO. 1 to B16F10 cells.

[0074] Figure 5 shows the results of confirming the expression levels of melanosome migration-related genes Rab27a, MYO5A, and MLPH by RT-PCR after adding a peptide consisting of the amino acid sequence of SEQ ID NO. 1 to B16F10 cells.

[0075] Figure 6 shows the results of confirming the expression levels of Rab27a and MLPH, which are melanosome migration-related genes, through Western blot after adding a peptide consisting of the amino acid sequence of SEQ ID NO. 1 to B16F10 cells.

[0076] The present invention will be explained in more detail below through examples. However, these examples are intended to illustrate the invention and the scope of the invention is not limited to these examples.

[0077] Example 1. Synthesis of Peptides

[0078] Peptides having the amino acid sequence of SEQ ID NO. 1 listed in [Table 1] below were synthesized using an automated peptide synthesizer (Milligen 9050, Millipore, USA), and these synthesized peptides were purified using C18 reverse-phase high-performance liquid chromatography (HPLC) (Waters Associates, USA). An ACQUITY UPLC BEH300 C18 column (2.1 mm x 100 mm, 1.7 µm, Waters Co, USA) was used.

[0079] [Table 1]

[0080]

[0081] Example 2. Confirmation of melanin production inhibitory effect

[0082] 7×10 mouse melanoma cells were placed in a 6-well plate 4After seeding at a density of cells / well, the cells were cultured for 24 hours. Subsequently, the culture medium was replaced with a medium containing 2% FBS, and 200 ng / ml of α-MSH (α-melanocyte-stimulating hormones) was added to promote melanin production. Along with this, each peptide consisting of the amino acid sequence of SEQ ID NO. 1 was added at concentrations of 50 μM, 100 μM, or 200 μM, and the cells were cultured in a CO2 incubator at 37°C for 72 hours. Afterward, the cultured cells were lysed with 1N NaOH, and the absorbance at 450 nm was measured. Meanwhile, an untreated group (Con) was used as a control, a group with only α-MSH added was used as a negative control, and a group with Arbutin (500 μM) and α-MSH added was used as a positive control.

[0083] As a result, as shown in Figure 1, it was confirmed that the production of melanin was reduced by the addition of a peptide consisting of the amino acid sequence of SEQ ID NO. 1.

[0084] Example 3. Confirmation of melanin expression inhibitory effect

[0085] 3×10⁶ mouse melanoma cells were placed in a 6-well plate 5After seeding at a density of cells / well, the cells were cultured for 24 hours. Subsequently, the culture medium was replaced with serum-free media and cultured for 24 hours. To promote melanin production, 200 ng / ml of α-MSH was added, along with the addition of each peptide consisting of the amino acid sequence of SEQ ID NO. 1 at concentrations of 50 μM, 100 μM, or 200 μM, and the cells were cultured for 10 minutes. Subsequently, after lysing the cultured cells, Western blot was performed using phospho-CREB antibody (cell signaling (Danvers, MA, USA), #9198) and actin antibody (santa cruz (Dallas, TX, USA), sc-47778) as primary antibodies, and Peroxidase-AffiniPure Goat Anti-Mouse IgG (H+L) antibody (Jackson ImmunoResearch (West Grove, PA, USA), 115-035-003) as secondary antibodies. In addition, the relative expression levels were quantified by calculating the p-CREB band densitometry results relative to the actin band densitometry results and quantifying each result based on the control group results. Meanwhile, an untreated group was used as the control, a group with only α-MSH added was used as the negative control, and a group with Arbutin (500 μM) and α-MSH added was used as the positive control.

[0086] As a result, as shown in Figure 2, it was confirmed that the phosphorylation level of CREB increased by α-MSH was inhibited by the addition of a peptide consisting of the amino acid sequence of SEQ ID NO. 1. Since CREB is a regulator of MITF expression, a major transcription factor in melanin production, these experimental results indicate that treatment with the peptide according to one embodiment can inhibit the expression of melanin.

[0087] Example 4. Confirmation of inhibitory effect on melanin production-related factors

[0088] 4-1. Evaluation of Expression of Melanin Production-Related Genes

[0089] 5×10 mouse melanoma cells were placed in a 6-well plate. 4 After seeding at a density of cells / well, the cells were cultured for 24 hours. Subsequently, the culture medium was replaced with a medium containing 2% FBS, and 200 ng / ml of α-MSH was added to promote melanin production. Along with this, each peptide consisting of the amino acid sequence of SEQ ID NO. 1 was added at concentrations of 50 μM, 100 μM, or 200 μM, and the cells were cultured for 72 hours at 37°C in a CO2 incubator. After extracting mRNA from the cultured cells, each cDNA was synthesized by reverse transcribing the extracted mRNA using a cDNA synthesis kit & PCR pre-mix (Intron, Korea). Subsequently, a polymerase chain reaction was performed using the cDNA and MITF, TYR, and TYRP1 primers. In addition, the band densitometry results of each marker were calculated relative to the GAPDH band densitometry results, and the relative expression levels were quantified by quantifying each result based on the control group results. Meanwhile, an untreated group was used as the control group, a group with only α-MSH added was used as the negative control group, and a group with Arbutin (500 μM) and α-MSH added was used as the positive control group.

[0090] [Table 2]

[0091]

[0092] As a result, as shown in FIG. 3, it was confirmed that the expression levels of MITF, TYR, and TYRP1, which were increased by α-MSH, were inhibited by the addition of a peptide consisting of the amino acid sequence of SEQ ID NO. 1. Since the MITF, TYR, and TYRP1 genes are factors related to melanin production, these experimental results indicate that treatment with the peptide according to one embodiment can inhibit the production of melanin.

[0093] 4-2. Evaluation of Melanin Production-Related Protein Expression

[0094] 7×10 mouse melanoma cells were placed in a 6-well plate 4After seeding at a density of cells / well, the cells were cultured for 24 hours. Subsequently, the culture medium was replaced with a medium containing 2% FBS, and 200 ng / ml of α-MSH was added to promote melanin production. Along with this, each peptide consisting of the amino acid sequence of SEQ ID NO. 1 was added at 50 μM, 100 μM, or 200 μM, and the cells were cultured for 72 hours. Subsequently, after lysing the cultured cells, Western blot was performed using MITF antibody (santa cruz (Dallas, TX, USA), sc-56433), TYR antibody (santa cruz (Dallas, TX, USA), sc-20035), TYRP1 antibody (santa cruz (Dallas, TX, USA), sc-166857), and actin antibody (santa cruz (Dallas, TX, USA), sc-47778) as primary antibodies, and Peroxidase-AffiniPure Goat Anti-Mouse IgG (H+L) antibody (Jackson ImmunoResearch (West Grove, PA, USA), 115-035-003) as secondary antibodies. In addition, the band densitometry results of each marker were calculated relative to the actin band densitometry results, and the relative expression levels were quantified by quantifying each result based on the control group results. Meanwhile, an untreated group was used as the control group, a group with only α-MSH added as the negative control group, and a group with Arbutin (500 μM) and α-MSH added as the positive control group.

[0095] As a result, as shown in FIG. 4, it was confirmed that the expression levels of MITF, TYR, and TYRP1, which were increased by α-MSH, were inhibited by the addition of a peptide consisting of the amino acid sequence of SEQ ID NO. 1. Since MITF, TYR, and TYRP1 are factors related to melanin production, these experimental results indicate that treatment with the peptide according to one embodiment can inhibit the production of melanin.

[0096] Example 5. Confirmation of inhibitory effect on factors related to melanosome migration

[0097] 5-1. Evaluation of Expression of Genes Related to Melanosomal Migration

[0098] 5×10 mouse melanoma cells were placed in a 6-well plate. 4After seeding at a density of cells / well, the cells were cultured for 24 hours. Subsequently, the culture medium was replaced with a medium containing 2% FBS, and 200 ng / ml of α-MSH was added to promote melanin production. Along with this, each peptide consisting of the amino acid sequence of SEQ ID NO. 1 was added at concentrations of 50 μM, 100 μM, or 200 μM, and the cells were cultured for 72 hours at 37°C in a CO2 incubator. After extracting mRNA from the cultured cells, each cDNA was synthesized by reverse transcribing the extracted mRNA using a cDNA synthesis kit & PCR pre-mix (Intron, Korea). Subsequently, a polymerase chain reaction was performed using the cDNA and Rab27a, MYO5A, and MLPH primers. In addition, the band densitometry results of each marker were calculated relative to the GAPDH band densitometry results, and the relative expression levels were quantified by quantifying each result based on the control group results. Meanwhile, an untreated group was used as the control group, a group with only α-MSH added was used as the negative control group, and a group with Arbutin (500 μM) and α-MSH added was used as the positive control group.

[0099] [Table 3]

[0100]

[0101] As a result, as shown in Figure 5, it was confirmed that the expression levels of Rab27a, MYO5A, and MLPH, which were increased by α-MSH, were inhibited by the addition of a peptide consisting of the amino acid sequence of SEQ ID NO. 1. Since the Rab27a, MYO5A, and MLPH genes are factors related to melanosome migration, these experimental results indicate that treatment with the peptide according to one embodiment can inhibit melanosome migration.

[0102] 5-2. Evaluation of Expression of Melanosome Migration-Related Proteins

[0103] 7×10 mouse melanoma cells were placed in a 6-well plate 4 After seeding at a density of cells / well, the cells were cultured for 24 hours. Subsequently, the culture medium was replaced with a medium containing 2% FBS, and 200 ng / ml of α-MSH was added to promote melanin production. Along with this, each peptide consisting of the amino acid sequence of SEQ ID NO. 1 was added at 50 μM, 100 μM, or 200 μM, and the cells were cultured for 72 hours. Subsequently, after lysing the cultured cells, Western blot was performed using Melanophilin antibody (abcam (Cambridge, Cambridgeshire, UK), ab2716), RAB27A antibody (abcam (Cambridge, Cambridgeshire, UK), ab55667), and actin antibody (santa cruz (Dallas, TX, USA), sc-47778) as primary antibodies, and Peroxidase-AffiniPure Goat Anti-Mouse IgG (H+L) antibody (Jackson ImmunoResearch (West Grove, PA, USA), 115-035-003) and Peroxidase-AffiniPure Rabbit Anti-Goat IgG (H+L) antibody (Jackson ImmunoResearch (West Grove, PA, USA), 305-035-003) as secondary antibodies. In addition, the band densitometry results of each marker were calculated relative to the actin band densitometry results, and the relative expression levels were quantified by quantifying each result based on the control group results. Meanwhile, an untreated group was used as the control group, a group with only α-MSH added was used as the negative control group, and a group with Arbutin (500 μM) and α-MSH added was used as the positive control group.

[0104] As a result, as shown in Fig. 6, it was confirmed that the expression levels of Rab27a and MLPH, which are increased by α-MSH, were inhibited by the addition of a peptide consisting of the amino acid sequence of SEQ ID NO. 1. Since Rab27a and MLPH are factors related to melanosome migration, these experimental results indicate that treatment with the peptide according to one embodiment can inhibit the migration of melanosomes.

[0105] Based on the above experimental results, it can be seen that the peptide composed of the amino acid sequence of SEQ ID NO. 1 according to one embodiment exhibits skin whitening efficacy by inhibiting melanin production and inhibiting melanosome migration.

[0106] Formulation Example 1. Preparation of peptide nanosomes

[0107] 50 mg of the peptide from Example 1 was dissolved by stirring thoroughly in 500 ml of distilled water. The mixture solution was mixed with 5 g of lecithin, 0.3 ml of sodium oleate, 50 ml of ethanol, and a small amount of oil, and then the volume was adjusted with distilled water to make the total volume 1 L. The mixture was then emulsified using a microfluidizer under high pressure to produce peptide nanosomes with a size of approximately 100 nm.

[0108] Formulation Example 2. Softening lotion

[0109] A softening lotion containing a peptide according to one embodiment and composed of the following composition was prepared by employing a method known in the art.

[0110] [Table 4]

[0111]

[0112] Formulation Example 3. Nourishing Cream

[0113] A nourishing cream comprising a peptide according to one embodiment and having the following composition was prepared by employing a method known in the art.

[0114] [Table 5]

[0115]

[0116] Formulation Example 4. Nourishing lotion

[0117] A nourishing cosmetic water comprising a peptide according to one embodiment and having the following composition was prepared by employing a method known in the art.

[0118] [Table 6]

[0119]

[0120] Formulation Example 5. Essence

[0121] An essence comprising a peptide according to one embodiment and having the following composition was prepared by employing a method known in the art.

[0122] [Table 7]

[0123]

[0124] The foregoing description of the present invention is for illustrative purposes only, and those skilled in the art will understand that other specific forms can be easily modified without altering the technical spirit or essential features of the present invention. Therefore, the embodiments described above should be understood as illustrative in all respects and not restrictive.

Claims

1. A peptide consisting of the amino acid sequence of sequence number 1.

2. The peptide of claim 1, wherein the N-terminus of the peptide is bonded to any one of the protecting groups 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 polyethylene glycol (PEG).

3. The peptide of claim 1, wherein the C-terminus of the peptide is bonded to any one of the protecting groups selected from the group consisting of an amino group (-NH2), a tertiary alkyl group, and a hydrazino group (-NHNH2).

4. The peptide of claim 1, wherein the peptide exhibits one or more characteristics selected from the following: (a) Inhibition of melanin production; (b) Inhibition of CREB (cAMP response element-binding protein) phosphorylation; (c) inhibition of MITF (Microphthalmia-associated transcription factor), TYR (tyrosinase), or TYRP1 (tyrosinase-related protein 1) expression; and (d) Inhibition of Rab27a, MYO5A (myosin 5a), or MLPH (Melanophilin) ​​expression.

5. A skin whitening composition comprising the peptide of any one of claims 1 to 4 as an active ingredient.

6. A cosmetic composition comprising the skin whitening composition of Claim 5.

7. A pharmaceutical composition for the prevention or treatment of hypermelanin pigmentation disorders comprising the skin whitening composition of Claim 5.

8. A pharmaceutical composition according to claim 7, wherein the melanin hyperpigmentation disorder is melasma, freckles, senile pigment spots, or solar lentigines.

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