KR20180089440A - Melanin inhibiting functional peptide and composition comprising same - Google Patents

Abstract

The present invention relates to a novel peptide showing skin whitening activity and a skin whitening composition containing the same, and more particularly to a novel peptide having an effect of inhibiting intracellular melanin production and a skin whitening composition containing the same. The novel peptide according to the present invention and the composition containing the novel peptide exhibit an excellent effect in inhibiting melanin production, and can provide a composition for improving skin color, a composition for skin whitening, and a method for treating skin whitening.

Description

Melanin inhibiting functional peptide and composition comprising same

The present invention relates to a melanin inhibitory functional peptide and a composition containing the melanin inhibitory functional peptide, and more particularly, to a functional peptide composition containing a melanin inhibitory functional peptide and effective for skin color improvement and skin whitening by melanin production.

Skin color is influenced by vascular distribution, hemoglobin, thickness of the stratum corneum, carotene, melanin, etc. It is known that melanin distribution status and amount have a major influence. Therefore, it is important to demonstrate inhibition of melanin formation in order to verify the skin whitening effect.

Melanin is a pigment produced in melanocytes of skin epidermis, which determines skin color and protects skin from damage caused by ultraviolet rays. Melanin is produced by a nonenzymatic oxidation reaction after tyrosine, an amino acid, is converted to DOPA or dopaquinone by an enzyme called tyrosinase. Accordingly, arbutin, which is a kind of tyrosinase inhibitor that inhibits the function of tyrosinase, has been developed and used.

In addition, melanin is produced in specific cells called melanocytes in the basal layer of skin, and the amount of melanin produced is determined by the activity of these cells. One of the most important factors affecting the activity of melanocytes is the α-melanocyte stimulating hormone (α-MSH). α-MSH is a hormone that directly affects melanocyte growth and melanin biosynthesis activity. It has been known that α-MSH is produced in the pituitary gland and affects the skin through blood vessels. However, α-MSH has been known to stimulate skin cells (melanocytes, Site) and has been found to play an important role in the blackening of the skin. It has also been reported that α-MSH is administered to humans in the presence of a large amount of blood vessels, and melanocyte production increases with α-MSH administration to melanocyte cells isolated and cultured from various races. .

In order to achieve skin whitening through inhibition of melanin production, a substance having an inhibitory activity against tyrosinase such as hydroquinone, ascorbic acid, kojic acid, glutathione is ointment, essence When used in combination with cosmetics, such as spots, freckles and skin hypercholesterolemia was improved. However, although hydroquinone exerts a predetermined whitening effect, there is a problem that the skin irritation is so severe that the compounding amount must be limited to a very small amount. Ascorbic acid is easily oxidized and the cosmetic composition containing the hydroquinone is discolored or deteriorated. . Furthermore, thiol compounds such as glutathione and cysteine have not only unpleasant odors but also problems with transdermal absorption, and their glycosides and derivatives are also highly polar, making them difficult to use as a blending ingredient for cosmetics. On the other hand, the placenta extract and the like have no irritation to the skin, but the whitening effect is insufficient.

Therefore, existing whitening substances have problems in terms of stability, such as skin allergy and toxicity. Generally, cosmetics are used daily for healthy skin. Safety should be given the highest priority and side effects should not occur. There is a need for research on melanin inhibitors that are not available.

Peptides are known to have a higher molecular weight, a higher production cost, and a decreased cell permeability, as a function of length. Thus, studies using peptides for skin whitening tend not to be actively studied. However, peptides have little or no tendency to cause toxicity and adverse effects compared with conventional chemical whitening substances, and, as is generally known, peptides exhibit excellent cell permeability and exhibit whitening effects even at a small dose. And a new whitening agent is being developed.

WO 2011-126163 A1 KR 10-2002-0016298 A

Under these circumstances, the inventors of the present invention have developed peptides showing new activity and found that they have an effect such as skin whitening through melanin inhibition, and completed the present invention.

It is an object of the present invention to provide a novel peptide having a skin whitening effect through melanin inhibition and a skin whitening composition containing the novel peptide.

One aspect of the present invention relates to a peptide comprising an amino acid sequence represented by one of SEQ ID NOS: 1 to 28, a peptide which is a fragment of the peptide sequence, a peptide having 80% or more sequence homology with the peptide sequence or a fragment thereof, Or a salt thereof, which exhibits skin whitening activity selected from the group consisting of < RTI ID = 0.0 >

According to one aspect of the present invention, the peptide is selected from the group consisting of SEQ ID NOS: 6, 12, 16, 17, 18, 19, 20, 21, , SEQ ID NO: 25, SEQ ID NO: 26, or the like.

According to an aspect of the present invention, the peptide may be at least one amino acid sequence selected from the group consisting of SEQ ID NO: 12, SEQ ID NO: 16, SEQ ID NO: 17 and SEQ ID NO:

According to an aspect of the present invention, the peptide may be any one or more amino acid sequences selected from the group consisting of SEQ ID NO: 6, SEQ ID NO: 8 and SEQ ID NO: 10.

According to one aspect of the present invention, the peptide may be characterized by inhibiting melanin production.

Another aspect of the present invention provides an external preparation for skin comprising the peptide or a salt thereof.

Another aspect of the present invention provides a cosmetic composition comprising the peptide or a salt thereof.

According to another aspect of the present invention, the composition comprises the peptide or a salt thereof in an amount of 1 μg / L to 10 g / L, preferably 0.0001 mg / mL to 10 mg / mL, more preferably 0.001 mg / mL 5 mg / mL, more preferably 0.01 mg / mL to 0.5 mg / mL.

According to another aspect of the present invention, the cosmetic composition may be any one selected from the group consisting of a skin lotion, a cream, a foundation, an essence, a gel, a pack, a foam cleansing, a soap and a skin external ointment .

Another aspect of the present invention provides a composition for a health functional food comprising the peptide or a salt thereof.

Another aspect of the present invention provides a pharmaceutical composition for preventing and treating diseases caused by melanin overgrowth, comprising the peptide or a salt thereof.

Another aspect of the present invention provides a skin whitening method comprising administering a composition comprising the peptide or a salt thereof to a subject in need of skin whitening.

Another aspect of the present invention relates to a method of preventing or treating diseases caused by melanin overgrowth, comprising administering to a subject in need thereof a composition comprising the peptide or a salt thereof, Prevention or treatment.

Another aspect of the present invention relates to a composition comprising a peptide or a salt thereof; And a kit for use for skin color improvement and skin whitening treatment effects, including instructions for starting at least one of the usage amount, the use method, and the use frequency of the composition.

Another aspect of the present invention provides the use of the peptide or a salt thereof for the preparation of a composition for skin color improvement and skin whitening.

The peptides having the sequence of SEQ ID NO: 1 according to the present invention have a melanin inhibitory effect and are expected to provide a composition for skin whitening through melanin inhibition and a method for treating skin whitening using the same.

1 is a graph showing the results of immunoprecipitation of a mouse skin cancer cell line (B16F1) cultured cells with a positive control group treated with α-MSH and a conventional whitening substance arbutin, and a peptide CP-1 according to the present invention after treatment with α- CP-2, CP-4, CP-5, CP-6, CP-7, CP-9, CP-11, CP- 18, CP-19, CP-20, CP-21, CP-22, CP-23, CP-24, CP-25, CP- CP-31 and CP-43, respectively.
FIG. 2 is a graph showing the results of treatment of a normal control group (con), a negative control group treated with melanin-producing inducible a-MSH, and a conventional whitening substance arbutin after treatment with? -MSH in a mouse skin cancer cell line (B16F1) A positive control group, and α-MSH treated peptides CP-1 to CP-10, respectively, and the results are shown in the graphs showing relative amounts (%) of melanin production based on a reference control group.
FIG. 3 is a graph showing the results of a comparison between the control group (con) in which no treatment was performed, the negative control treated with melanin-producing inducible hormone a-MSH, the conventional whitening substance arbutin after treatment with alpha-MSH in the mouse skin cancer cell line (B16F1) A positive control group, and α-MSH treated peptides CP-11 to CP-19, respectively, and the results are shown in graphs showing relative amounts (%) of melanin production based on a reference control group.
FIG. 4 is a graph showing the results of a comparison between a reference control (con) in which no treatment was performed, a negative control in which melanin-producing inducible hormone a-MSH was treated, α-MSH in the mouse skin cancer cell line (B16F1) A positive control group, and α-MSH-treated experimental groups treated with peptides CP-20 to CP-31, respectively, and graphs showing relative amounts of melanin production as a reference control group (%).
FIG. 5 is a graph showing the results of a comparison between a reference control (con) treated with nothing, a negative control treated with melanin-producing inducible a-MSH, and a conventional whitening substance arbutin after treatment with? -MSH in mouse skin cancer cell line (B16F1) A positive control group, and an experimental group treated with peptides CP-32 to CP-43, respectively, after the treatment with? -MSH, and the relative amounts (%) of melanin production as a reference control group.
FIG. 6 is a graph showing the results of a comparison between a reference control (con) in which no treatment was performed, a negative control treated with melanin-producing inducible a-MSH, α-MSH and a conventional whitening substance arbutin after treatment in mouse skin cancer cell line (B16F1) (%) On the basis of the reference control group when the test group was divided into experimental groups treated with peptides CP-44 to CP-53 after treatment with a positive control group and α-MSH.

The present invention can be variously modified and can have various embodiments, and the present invention will be described in more detail as follows. It is to be understood, however, that the invention is not to be limited to the specific embodiments, but includes all modifications, equivalents, and alternatives falling within the spirit and scope of the invention. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS Hereinafter, the present invention will be described in detail with reference to the accompanying drawings.

In one aspect of the invention, there is disclosed a novel peptide consisting of a sequence of SEQ ID NO: 1 to SEQ ID NO: 28.

The peptides disclosed herein may comprise at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% of the novel peptides of SEQ ID NOS: And may include peptides having sequence homology. The peptides disclosed in the present specification can also be produced by reacting peptides represented by one of SEQ ID NOS: 1 to 28 with one or more amino acids, two or more amino acids, three or more amino acids, four or more amino acids, five or more amino acids, And may include peptides in which 7 or more amino acids have been changed.

In one aspect of the invention, the amino acid change is of a property that causes the physicochemical properties of the peptide to change. For example, amino acid changes such as improving the thermal stability of the peptide, altering the substrate specificity, changing the optimum pH, etc. can be performed.

The peptides of SEQ ID NOS: 1 to 28 according to one aspect of the present invention have an advantage of low intracellular toxicity and high in vivo stability. The peptides shown in SEQ ID NOS: 1 to 28 are shown in Table 1 below. The "name" in Table 1 below is what the peptides are named for.

SEQ ID NO. name order Length One CP-1 PALLTSRLR 9 2 CP-2 LLTSRLR 7 3 CP-4 RPALLTSR 8 4 CP-5 PALLTSR 7 5 CP-6 ALLTSR 6 6 CP-7 ALLTS 5 7 CP-9 RPALLTS 7 8 CP-11 RFI 3 9 CP-13 LLTSRLRF 8 10 CP-14 LRF 3 11 CP-15 RLRFIPK 7 12 CP-16 TSRLRFIP 8 13 CP-17 ARPALLTSR 9 14 CP-18 SRLRFIP 7 15 CP-19 TSRLRFI 7 16 CP-20 ALSSRLRA 8 17 CP-21 ALSSRLRG 8 18 CP-22 ALSSRLRF 8 19 CP-23 ALSTRLRA 8 20 CP-24 ALSTRLRG 8 21 CP-25 ALSTRLRF 8 22 CP-26 ALTSRVRA 8 23 CP-27 ALTSRVRG 8 24 CP-28 ALTSRVRF 8 25 CP-29 ALTSCLA 8 26 CP-30 ALTSKLRG 8 27 CP-31 ALTSKLRF 8 28 CP-43 RPALLTSR 8

In one aspect of the present invention, there is provided a composition comprising at least one peptide comprising the amino acid sequence of any one of SEQ ID NOS: 1 to 28 as an active ingredient.

The whitening composition according to one aspect of the present invention comprises, in one aspect, 0.0001 mg / mL to 10 mg / mL, more specifically 0.001 mg / mL to 5 mg / mL of each peptide of one of SEQ ID NOs: May be contained in an amount of 0.01 mg / mL to 0.5 mg / mL, but it is possible to appropriately control the difference in the effect depending on the dose. When it is contained in the above-mentioned range or below, it is not only suitable for exhibiting the intended effect of the present invention but also can satisfy both the stability and safety of the composition and may be suitably included in the above range in terms of cost effectiveness .

The composition according to one aspect of the present invention can be applied to all animals including humans, dogs, chickens, pigs, cattle, sheep, guinea pigs or monkeys.

The composition for external application for skin according to one aspect of the present invention is not particularly limited in its formulation and may be applied to various cosmetic compositions such as a softening agent, a convergent lotion, a nutritional lotion, an eye cream, a nutritional cream, a massage cream, a cleansing cream, a cleansing foam, a cleansing water, Essence, pack, and the like.

The cosmetic composition according to one aspect of the present invention can be manufactured in various forms according to a conventional method for producing a cosmetic. For example, the cosmetic composition may be prepared in the form of a perfumed product containing the peptide, a lotion, a cream, a lotion or the like, and may be diluted with a conventional cleansing solution, a converging solution and a moisturizing solution. The cosmetic composition may also contain conventional adjuvants such as stabilizers, solubilizers, vitamins, pigments, and flavorings commonly used in the cosmetic composition arts.

In addition, the peptide having skin whitening activity according to one aspect of the present invention antagonizes the action of? -MSH (? -Milanocyte stimulating hormone), and thus, for example, a pharmaceutically acceptable carrier, excipient, -MSH-related diseases, as well as to be used in cosmetic preparations such as skin, lotion, cream, foundation, essence, gel, pack, foam cleansing, Addition to the composition for skin whitening can exhibit a strong skin whitening effect without any adverse side effects.

Formulations for oral administration of a composition according to one aspect of the present invention may be, but are not limited to, tablets, pills, soft or hard capsules, granules, powders, solutions or emulsions.

The composition according to one aspect of the present invention may contain additives such as diluents, excipients, lubricants, binders, disintegrants, buffers, dispersants, surfactants, colorants, fragrances or sweeteners as needed. The composition according to one aspect of the present invention can be prepared by a conventional method in the art.

The composition according to one aspect of the present invention can inhibit the formation of intracellular melanin to improve skin pigmentation phenomena such as dullness, freckles, spots, etc., and can alleviate skin yellowing and redness, thereby improving skin brightness and uniformity.

The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. The abbreviated term in front of a noun is not intended to limit the quantity but to indicate that there is more than one noun item mentioned. The terms " comprising ", " having ", and " containing " are to be construed as open (i.e., meaning including but not limited to).

To refer to a range of values is an easy way to substitute for referring individually to each distinct value falling within that range and each separate value that is not explicitly stated is referred to individually in the specification Are incorporated herein by reference. All range end values are contained within that range and can be combined independently.

All methods mentioned herein may be performed in any suitable order unless otherwise indicated or clearly contradicted by context. It is to be understood that the use of any embodiment and all of the embodiments or example language (e.g., " such as ") is for the purpose of describing the present invention only, It is not intended to be limiting. No language in the specification should be construed as obliging any non-claimed components to practice the present invention. Unless defined otherwise, the technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

Preferred embodiments of the present invention include the most optimal mode known to the inventors for carrying out the present invention. Variations of the preferred embodiments may become apparent to those skilled in the art upon reading the foregoing description. The inventors expect those skilled in the art to appropriately utilize such variations, and the inventors expect the invention to be practiced otherwise than as described herein. Accordingly, the present invention includes equivalents and all modifications of the subject matter of the invention as recited in the appended claims, as permitted by the patent law. Moreover, any combination of the above-mentioned components within all possible variations is included in the present invention unless otherwise specified or contradicted by context. While the present invention has been particularly shown and described with reference to exemplary embodiments, those skilled in the art will readily appreciate that many changes can be made in form and detail without departing from the spirit and scope of the invention as defined by the following claims .

Hereinafter, the constitution and effects of the present invention will be described in more detail with reference to Examples and Experimental Examples. However, the following examples and experimental examples are provided for illustrative purposes only in order to facilitate understanding of the present invention, and the scope and scope of the present invention are not limited thereto.

Example 1: Synthesis of novel peptides

A novel peptide (hereinafter referred to as " CP-1 to CP-53 ") was prepared according to a conventionally known solid phase peptide synthesis method. Specifically, the peptides were synthesized by coupling one amino acid from the C-terminal through Fmoc solid phase peptide synthesis (SPPS) using ASP48S (Peptron, Inc., Daejeon, Korea). The first amino acid at the C-terminus of the peptides attached to the resin was used as follows. For example:

NH ₂ -Lys (Boc) -2-chloro-Trityl Resin

NH ₂ -Ala-2-chloro-Trityl Resin

NH ₂ -Arg (Pbf) -2-chloro-Trityl Resin

Boc, t-Bu (t-butylester), Pbf (2, 2, 4, 6, 6), which are all amino acid sources used for peptide synthesis, are protected by N-term and Fmoc, 7-pentamethyl dihydro-benzofuran-5-sulfonyl). For example:

Fmoc-Ala-OH, Fmoc-Arg (Pbf) -OH, Fmoc-Glu (OtBu) -OH, Fmoc-Pro-OH, Fmoc-Leu-OH

As the coupling reagent, HBTU [2- (1H-Benzotriazole-1-yl) -1,1,3,3-tetramethylaminium hexafluorophosphate] / HOBt [N-Hydroxxybenzotriazole] / NMM [4-Methylmorpholine] Respectively. Fmoc removal was performed using piperidine in DMF in 20% DMF. Cleavage Cocktail [TFA (trifluoroacetic acid) / TIS (triisopropylsilane) / EDT (ethanedithiol) / H ₂ O = 92.5 / 2.5 / 2.5 / 2.5] was used to remove the synthesized peptide from Resin and to remove the protecting group of the residue. Respectively.

Each of the peptides was synthesized by repeating the steps of reacting corresponding amino acids with a starting amino acid having an amino acid protecting group bonded thereto on a solid support, washing with a solvent, followed by deprotection. The synthesized peptide was cleaved from the resin and purified by HPLC. The success of the synthesis was confirmed by LC / MS and lyophilized.

As a result of high performance liquid chromatography on the peptides used in this example, the purity of all the peptides was 95% or more.

For example, a specific procedure for preparing peptide CP-16 will be described as follows.

1) Coupling

NH _2-protected amino acid to the -A-2-chloro-Trityl Resin (8 equivalents) and the coupling reagent HBTU (8 eq.) / HOBt (8 eq.) / NMM (16 equivalents) was added dissolved in DMF, at room temperature The reaction was carried out for 2 hours and washed sequentially with DMF, MeOH and DMF.

2) Fmoc deprotection

Piperidine in DMF in 20% DMF was added, and the reaction was carried out at room temperature for 5 minutes twice, followed by washing with DMF, MeOH and DMF.

3) The reaction of 1 and 2 was repeated to make the peptide basic backbone NH ₂ -T (tBu) -S (tBu) -R (Pbf) -LR (Pbf) -FIP-2-chloro-Trityl Resin.

4) Cleavage: Cleavage cocktail was added to the synthesized peptide Resin to separate the peptide from Resin.

5) Add Cooling diethyl ether to the obtained mixture and centrifuge to precipitate the obtained peptide.

6) The crude peptides obtained in the above procedure were separated and purified using Prep HPLC. The column was a Vydac Everest C18 column (250 mm × 22 mm, 10 μm). Eluent was a water-acetonitrile linear gradient (10-75% (v / v) of acetonitrile) containing 0.1% (v / v) trifluoroacetic acid.

7) Whether the separated peptides were synthesized according to the desired sequence was confirmed by LC / MS (Agilent HP1100 series).

8) The peptides with molecular weight confirmed were purified by analytical HPLC at 95% purity or higher, and lyophilized to prepare white powder.

52 peptides of peptides CP-1 to CP15 and CP-17 to CP-53 were prepared by substituting only the basic skeleton of the peptide to the corresponding sequence of each peptide based on the same method as that of CP-16.

Example 2: Intracellular melanin formation inhibitory activity of novel peptides

In order to examine the inhibitory activity on the intracellular melanin production of the novel peptide, the mouse skin cancer cell line (B16F1) was cultured and treated with the same concentration of each of the peptides (CP-1 to CP-53) Experiments were carried out to measure the amount through absorbance.

Experimental cell culture

A mouse skin cancer cell line (B16F1, KCLB number: 80007) was used for the experiment. The cells were seeded in a 100 cm ² culture dish with DMEM supplemented with 10% FBS and penicillin-streptomycin antibiotics, and incubated in an incubator (37 ° C, 5% CO ₂ ) Lt; / RTI > Cells were maintained in good condition by sub-culture at 2 to 5 days intervals while observing the growth rate and condition of each cell.

The peptides CP-1 to CP-53 were synthesized according to the synthesis method in Example 1 using Peptron (Peptron, Daejeon, Korea).

Experimental Method

24 hours before, cancer cells were prepared and cultured in a 6-well plate at a level of 30-40% as mentioned in the culture method. The experiment was carried out using a negative control (α-MSH (0.4 μM) treated with only melanogenesis-inducing hormone α-MSH (0.4 μM), a reference control (no treatment other than the medium PBS, denoted as con in FIGS. 2 to 6) After treatment, a positive control group treated with arbutin (0.4 μM), known as a conventional whitening agent, treated with α-MSH (0.4 μM) and treated with peptides CP-1 to CP-53 at a concentration of 100 μM Respectively.

After 72 hours from the treatment of each group, the culture medium was removed, and the cells were recovered by washing with PBS and trypsin-EDTA enzyme treatment. The recovered cells were counted using a hemacytometer, centrifuged (7,500 rpm for 10 minutes), and the supernatant was removed to obtain a cell pellet. The cell pellet was dried at 60 ° C., and 100 μL of a 1 M sodium hydroxide solution containing 10% DMSO was added thereto to extract intracellular melanin in a 60 ° C. thermostatic chamber. The extracted solution was measured by absorbance at 490 nm with a microplate reader to calculate the amount of melanin in a given cell number.

Statistical processing

The amount of melanin produced in the control group (denoted by con), which had not been treated with the cells, was set to 100%, and the amount of melanin produced in the other control and experimental groups was relatively calculated. All data were expressed as mean ± standard error (mean ± SD) and statistical analysis was performed by t-test. Comparisons between the groups were made by the Tukey test method and it was judged that the p value was less than 0.05 in most peptides and statistically significant.

Experimental Results and Analysis

The results of measurement of inhibitory effect on intracellular melanin production derived from the above experimental method are as follows.

In the negative control group treated with melanin-inducing hormone a-MSH, the intracellular melanin production was increased about 2 times as compared to the reference control group (see FIGS. 2 to 6). In addition, melanin production is generally decreased in a positive control group treated with arbutin, which is a conventional whitening agent, in cells in which intracellular melanin is increased by? -MSH (see FIGS. 1 to 6).

In addition, the amount of melanin production in the experimental group treated with the peptides CP-1 to CP-53 after α-MSH treatment was decreased compared to the negative control (see FIGS. 2 to 6).

The inhibition rate (inhibition rate = 100% - (the amount of melanin production in the positive control group) was calculated by comparing the melanin production amount of the negative control group (? -MSH) and the melanin production amount of the positive control group treated with the conventional whitening agent arbutin Value / negative control melanin production amount) X100)% was 14.9%.

CP-1, CP-2, CP-4, CP-5, CP-6, CP-7, CP-9, CP-11 and CP- 13, CP-14, CP-15, CP-16, CP-17, CP-18, CP-19, CP-20, CP- (Inhibition rate = 100% - (the amount of melanin production in the experimental group) was calculated by comparing the production amount of the experimental group treated with CP-26, CP-27, CP-28, CP-29, CP-30, CP- Measurement value / measurement value of melanin production amount of negative control) X100)% is shown in the following Table 2 (see Fig. 1).

Peptides CP-3, CP-8, CP-10, CP-12, CP-32, CP-33, CP-34, CP-35, CP- , CP-41, CP-42, CP-44, CP-45, CP-46, CP-47, CP-48, The experimental group showed lower melanin production inhibitory effect than the arbutin-treated control group (14.9%), and was excluded from Table 2.

SEQ ID NO. name order Length % Inhibition One CP-1 PALLTSRLR 9 15.5 2 CP-2 LLTSRLR 7 14.9 3 CP-4 RPALLTSR 8 15.2 4 CP-5 PALLTSR 7 17.9 5 CP-6 ALLTSR 6 16.9 6 CP-7 ALLTS 5 24.5 7 CP-9 RPALLTS 7 16.3 8 CP-11 RFI 3 16.7 9 CP-13 LLTSRLRF 8 17.6 10 CP-14 LRF 3 19.2 11 CP-15 RLRFIPK 7 19.3 12 CP-16 TSRLRFIP 8 30.8 13 CP-17 ARPALLTSR 9 15.3 14 CP-18 SRLRFIP 7 18.3 15 CP-19 TSRLRFI 7 17.4 16 CP-20 ALSSRLRA 8 26.2 17 CP-21 ALSSRLRG 8 25.4 18 CP-22 ALSSRLRF 8 20.3 19 CP-23 ALSTRLRA 8 21.2 20 CP-24 ALSTRLRG 8 22 21 CP-25 ALSTRLRF 8 23.5 22 CP-26 ALTSRVRA 8 23.1 23 CP-27 ALTSRVRG 8 26.4 24 CP-28 ALTSRVRF 8 15 25 CP-29 ALTSCLA 8 22 26 CP-30 ALTSKLRG 8 23.7 27 CP-31 ALTSKLRF 8 17.4 28 CP-43 RPALLTSR 8 15.2

As shown in Table 2, peptides CP-1, CP-2, CP-4, CP-5, CP-6, CP-7, CP- 15, CP-16, CP-17, CP-18, CP-19, CP-20, CP-21, CP-22, CP- CP-28, CP-29, CP-30, CP-31 and CP-43 were shown to reduce melanin by more than 14.9% inhibition of melanogenesis in a positive control group treated with arbutin, a conventional whitening substance, In particular, in the case of CP-07, CP-16, CP-20, CP-21, CP-22, CP-23, CP-24, CP-25, CP-26, CP-27, (14.9% in the case of arbutin, a conventional whitening substance).

On the other hand, in order to confirm the stability as a cosmetic peptide, the stability of the peptide was confirmed by measuring the change in absorbance of the peptide after one to three days in a typical cosmetic solvent, which is typically used for each peptide. As a cosmetic solvent, 10% of 1,3-butyleneglycol (BG), 5.0% of ethanol (EtOH), 5.0% of Glycerin and 0.3% of Emalex-hc60 (surfactant) -11 is shown in Table 3 below.

Time 1,3 BG EtOH Glycerin Emalex60 0 h 7910899 7492353 8057268 8976881 24 h 7754934 7366026 7992905 8650562 72 h 7744398 7328666 8063024 8634648

As shown in Table 3, the change in absorbance of CP-11 in the four solvents was within ± 5%, indicating that CP-11 remained very stable as a cosmetic peptide.

In addition, in the additional stability test and skin toxicity test, the new peptides of Table 2 were found to be stable and non-toxic at room temperature. In particular, CP-7, CP-11 and CP-14 showed excellent melanin production inhibitory effect As well as safety and skin toxicity, thus being suitable for the preparation of compositions for skin whitening.

From the above experimental results, it can be seen that the novel peptides according to the present invention and compositions containing them show excellent melanin production inhibitory activity. It can be seen that the peptides according to the present invention and the compositions containing them can have an effect of inhibiting melanin production, which is a fundamental treatment of skin whitening.

The novel peptides according to the present invention have an effect of inhibiting intracellular melanin production and have no toxicity to the skin and are relatively stable at room temperature. As a result, it is possible to improve the skin surface, And cosmetics for skin whitening using the same.

<110> GemVax & KAEL., Co., LTD.          KIM, Sangjae <120> Peptides having melanin inhibition activity and the Composition          Comprising the Same <130> OF16P262 / PCT <150> KR 10-2015-0187946 <151> 2015-12-28 <160> 28 <170> KoPatentin 3.0 <210> 1 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> htrt derived peptide fragment <400> 1 Pro Ala Leu Leu Thr Ser Arg Leu Arg   1 5 <210> 2 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> htrt derived peptide fragment <400> 2 Leu Leu Thr Ser Arg Leu Arg   1 5 <210> 3 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> htrt derived peptide fragment <400> 3 Arg Pro Ala Leu Leu Thr Ser Arg   1 5 <210> 4 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> htrt derived peptide fragment <400> 4 Pro Ala Leu Leu Thr Ser Arg   1 5 <210> 5 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> htrt derived peptide fragment <400> 5 Ala Leu Leu Thr Ser Arg   1 5 <210> 6 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> htrt derived peptide fragment <400> 6 Ala Leu Leu Thr Ser   1 5 <210> 7 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> htrt derived peptide fragment <400> 7 Arg Pro Ala Leu Leu Thr Ser   1 5 <210> 8 <211> 3 <212> PRT <213> Artificial Sequence <220> <223> htrt derived peptide fragment <400> 8 Arg Phe Ile   One <210> 9 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> htrt derived peptide fragment <400> 9 Leu Leu Thr Ser Arg Leu Arg Phe   1 5 <210> 10 <211> 3 <212> PRT <213> Artificial Sequence <220> <223> htrt derived peptide fragment <400> 10 Leu Arg Phe   One <210> 11 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> htrt derived peptide fragment <400> 11 Arg Leu Arg Phe Ile Pro Lys   1 5 <210> 12 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> htrt derived peptide fragment <400> 12 Thr Ser Arg Leu Arg Phe Ile Pro   1 5 <210> 13 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> htrt derived peptide fragment <400> 13 Ala Arg Pro Ala Leu Leu Thr Ser Arg   1 5 <210> 14 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> htrt derived peptide fragment <400> 14 Ser Arg Leu Arg Phe Ile Pro   1 5 <210> 15 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> htrt derived peptide fragment <400> 15 Thr Ser Arg Leu Arg Phe Ile   1 5 <210> 16 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> htrt derived peptide fragment <400> 16 Ala Leu Ser Ser Arg Leu Arg Ala   1 5 <210> 17 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> htrt derived peptide fragment <400> 17 Ala Leu Ser Ser Arg Leu Arg Gly   1 5 <210> 18 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> htrt derived peptide fragment <400> 18 Ala Leu Ser Ser Arg Leu Arg Phe   1 5 <210> 19 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> htrt derived peptide fragment <400> 19 Ala Leu Ser Thr Arg Leu Arg Ala   1 5 <210> 20 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> htrt derived peptide fragment <400> 20 Ala Leu Ser Thr Arg Leu Arg Gly   1 5 <210> 21 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> htrt derived peptide fragment <400> 21 Ala Leu Ser Thr Arg Leu Arg Phe   1 5 <210> 22 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> htrt derived peptide fragment <400> 22 Ala Leu Thr Ser Arg Val Ala   1 5 <210> 23 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> htrt derived peptide fragment <400> 23 Ala Leu Thr Ser Arg Val Gly   1 5 <210> 24 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> htrt derived peptide fragment <400> 24 Ala Leu Thr Ser Arg Val Phe   1 5 <210> 25 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> htrt derived peptide fragment <400> 25 Ala Leu Thr Ser Lys Leu Arg Ala   1 5 <210> 26 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> htrt derived peptide fragment <400> 26 Ala Leu Thr Ser Lys Leu Arg Gly   1 5 <210> 27 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> htrt derived peptide fragment <400> 27 Ala Leu Thr Ser Lys Leu Arg Phe   1 5 <210> 28 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> htrt derived peptide fragment <400> 28 Arg Pro Ala Leu Leu Thr Ser Arg   1 5

Claims (15)

A peptide consisting of an amino acid sequence represented by any one of SEQ ID NOS: 1 to 28, a peptide which is a fragment of the peptide sequence, a peptide having a sequence homology of 80% or more with the peptide sequence or a fragment thereof A peptide or a salt thereof exhibiting skin whitening activity. The method of claim 1, wherein the peptide is selected from the group consisting of SEQ ID NO: 6, SEQ ID NO: 12, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: SEQ ID NO: 25, and SEQ ID NO: 26, or a salt thereof, wherein the peptide has at least one amino acid sequence selected from the group consisting of SEQ ID NO: The peptide or a salt thereof according to claim 1, wherein the peptide is at least one amino acid sequence selected from the group consisting of SEQ ID NO: 12, SEQ ID NO: 16, SEQ ID NO: 17 and SEQ ID NO: The peptide or a salt thereof according to claim 1, wherein the peptide is at least one amino acid sequence selected from the group consisting of SEQ ID NO: 6, SEQ ID NO: 8 and SEQ ID NO: 10. The peptide or its salt according to claim 1, wherein the peptide inhibits melanin production. An external preparation for skin comprising a peptide according to any one of claims 1 to 5, or a salt thereof. A cosmetic composition comprising a peptide according to any one of claims 1 to 5, or a salt thereof. 8. The composition of claim 7, wherein the composition comprises the peptide or a salt thereof in an amount of 0.0001 mg / mL to 10 mg / mL, preferably 0.001 mg / mL to 5 mg / mL, more preferably 0.01 mg / mL to 0.5 mg / mL By weight of the cosmetic composition. The cosmetic composition according to claim 8, wherein the cosmetic composition comprises any one of a skin, a lotion, a cream, a foundation, an essence, a gel, a pack, a foam cleansing, a soap and a skin external ointment. A composition for a health functional food comprising a peptide according to any one of claims 1 to 5, or a salt thereof. A pharmaceutical composition for preventing and treating diseases caused by melanin overgrowth, comprising a peptide according to any one of claims 1 to 5, or a salt thereof. A skin whitening method comprising administering to a subject in need of skin whitening a composition comprising a peptide according to any one of claims 1 to 5, or a salt thereof. 8. A method for preventing or treating melanoma, comprising administering to a subject in need thereof a composition comprising a peptide according to any one of claims 1 to 5, or a salt thereof, A method of preventing or treating a disease. A composition comprising a peptide according to any one of claims 1 to 5, or a salt thereof; And a kit for use for improving the skin color and skin whitening treatment, comprising an instruction that discloses at least one of the usage amount, the use method, and the use frequency of the composition. Use of a peptide according to any one of claims 1 to 5 or a salt thereof for the preparation of a composition for skin color improvement and skin whitening.

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