Peptides with skin condition improving activity and their uses

Abstract

The present application relates to a peptide having skin condition-improving activity and its uses, and provides a peptide consisting of the amino acid sequence of SEQ ID NO: 1, and a cosmetic composition for improving skin condition that contains the peptide consisting of the amino acid sequence of SEQ ID NO: 1 as an active ingredient.

Description

[Technical Field]

[0001] This application relates to a peptide having skin condition improving activity and its uses. [Background technology]

[0002] Human skin is constantly undergoing changes, the most representative of which are the decline in skin function and the decrease in visual beauty due to aging. Aging causes wrinkles in the skin, and typical factors contributing to wrinkle formation include ultraviolet radiation exposure and decreased collagen biosynthesis. Skin aging can be broadly divided into intrinsic aging, which is caused by genetic factors, and extrinsic aging, which is caused by external environmental factors such as sunlight. In the case of extrinsic aging, it is known that aging can be prevented, treated, or delayed through the removal of reactive oxygen species, proliferation of fibroblasts, and promotion of collagen biosynthesis.

[0003] On the other hand, collagen, a major component of the extracellular matrix, is the primary matrix protein produced by fibroblasts in the skin. Collagen forms most of the organic matter in the skin, tendons, bones, and teeth, but its content is particularly high in bones and skin (dermis). This collagen decreases with age and photoaging due to UV exposure, which is known to be closely related to the formation of wrinkles in the skin. Collagen also plays an important role in wound healing, promoting collagen synthesis in damaged epithelium, which can lead to rapid and scar-free healing of wounds. In addition, it has been reported that by promoting collagen biosynthesis, the density of the basal layer and other areas increases, which can lead to a lower melanin pigment concentration per unit skin density, resulting in a brighter skin tone.

[0004] Against this technological backdrop, multifaceted research is underway to improve skin conditions through mechanisms such as promoting collagen biosynthesis and increasing the proliferation and activity of fibroblasts (Korean Registered Patent No. 10-1813629), but the situation remains inadequate. [Overview of the project] [Problems that the invention aims to solve]

[0005] One embodiment is to provide a peptide consisting of the amino acid sequence of SEQ ID NO: 1.

[0006] Another embodiment provides a skin condition improving composition containing a peptide consisting of the amino acid sequence of SEQ ID NO: 1 as an active ingredient.

[0007] Other purposes and advantages of this application will be further clarified by the following detailed description, along with the claims and drawings. Any matters not described herein are readily apparent and can be inferred by any skilled person in the art of this application or a similar art, and therefore their description is omitted. [Means for solving the problem]

[0008] Each description and embodiment disclosed in this application may also apply 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 is not considered to be limited by the specific descriptions described below.

[0009] One embodiment provides a peptide consisting of the amino acid sequence of SEQ ID NO: 1.

[0010] As used herein, the term "peptide" may mean a linear molecule formed by the bonding of amino acid residues to one another by peptide bonds. Such peptides may be produced by chemical synthesis methods known to those skilled in the art, particularly by solid-phase or liquid-phase synthesis techniques (US Patent No. 5,516,891). The inventors, through diligent efforts to develop a peptide with biologically effective activity, have identified a peptide consisting of the amino acid sequence of Sequence ID No. 1. Herein, the biologically effective activity also includes exhibiting one or more of the following characteristics selected from: (a) promotion of fibroblast or keratinocyte proliferation; (b) enhancement of the expression of extracellular matrix components such as collagen type 1 (Col1a1), fibronectin, or elastin; (c) enhancement of the expression of skin barrier factors such as sirtuin-1 (SIRT-1) or aquaforin-3 (AQP3); and (d) inhibition of fibroblast or keratinocyte death and reduction of reactive oxygen species levels. Therefore, the peptide can be used for improving skin condition or for antioxidant purposes.

[0011] The peptide may also have a protecting group attached to its N or C terminus to acquire chemical stability, enhanced pharmacological properties (half-life, water absorption, potency, efficacy, etc.), altered specificity (e.g., broad biological activity spectrum), or reduced antigenicity. In one specific example, the N-terminus of the peptide is bonded to one protecting group selected from the group consisting of acetyl, fluorenylmethoxycarbonyl, formyl, palmitoyl, myristyl, stearyl, butoxycarbonyl, allyloxycarbonyl, and polyethylene glycol (PEG); and / or, the C-terminus of the peptide may be bonded to one protecting group selected from the group consisting of amino, -NH2, tertiary alkyl, and azide (-NHNH2). The peptide may also selectively further include targeted sequences, tags, labeled residues, and amino acid sequences manufactured for specific purposes to increase half-life or peptide stability.

[0012] The peptides are artificially synthesized, non-naturally occurring, or engineered, where “non-naturally occurring or engineered” means a state produced by artificial modification rather than the state of existence that occurs naturally. Here, the artificial modification may include artificially synthesizing an amino acid sequence by mimicking multiple amino acid structures, or being engineered to acquire chemical stability, enhanced pharmacological properties, altered specificity, or reduced antigenicity, as described above.

[0013] As used herein, the term "stability" may refer not only to in vivo stability, which protects the peptide from attack by endogenous protein-cleaving enzymes, but also to storage stability (e.g., room temperature storage stability).

[0014] Another embodiment provides a cosmetic composition for improving skin condition that contains a peptide consisting of the amino acid sequence of SEQ ID NO: 1 as an active ingredient.

[0015] As stated above, any terms or elements mentioned in the description of the peptides that are the same as those already mentioned are as described above.

[0016] As used herein, the term "improvement" may mean any action that alleviates or treats a condition, such as any action that at least reduces the severity of the symptoms.

[0017] As used herein, the term "improvement of skin condition" comprehensively refers to the process or effect of treating, reducing, or mitigating skin damage induced by intrinsic or extrinsic factors of the skin, and may, but is not limited to, wrinkle reduction, improved skin elasticity, wound healing, strengthening of the skin barrier, or inhibition of skin aging.

[0018] Here, "wrinkle improvement," "skin elasticity improvement," and "wound healing" can refer to all effects that increase the total amount of extracellular matrix factors, including the promotion of collagen synthesis. Furthermore, "strengthening the skin barrier" can refer to enhancing the skin's natural functions, such as preventing leakage of moisture and nutrients from the skin to the outside and preventing the penetration of harmful substances such as bacteria and viruses into the skin. Additionally, "suppression of skin aging" can refer to suppressing the decline of skin function, such as wrinkles, sagging, and loss of elasticity. In this context, the aforementioned skin aging also includes photoaging, such as skin aging caused by ultraviolet radiation.

[0019] Conventional functional peptides, despite their effective biological activity, suffer from disadvantages such as not being effectively absorbed into target tissues or cells due to their size, or being eliminated from the body quickly due to their short half-life. On the other hand, the cosmetic composition according to one embodiment contains a peptide consisting of 10 or fewer amino acids as an active ingredient, resulting in excellent skin penetration of the active ingredient. For example, when applied topically to the skin, it can effectively improve skin condition.

[0020] According to one embodiment, the peptide was able to promote the proliferation of fibroblasts or keratinocytes and enhance the synthesis of extracellular matrix components and skin barrier factors. Furthermore, it was shown that the peptide can restore the inhibited activity of fibroblasts and keratinocytes and enhance their antioxidant activity. Therefore, the peptide can be used as an active ingredient in cosmetic compositions for improving skin condition.

[0021] The cosmetic composition may, but is not limited to, contain a cosmetically effective amount of the peptide and / or a cosmetically acceptable carrier.

[0022] As used herein, the term "cosmetically effective amount" means an amount sufficient to achieve the skin condition improvement efficacy of the cosmetic composition.

[0023] The weight ratio between the peptide and the cosmetically acceptable carrier may be, for example, 500:1 to 1:500, and may also be, but is not limited to, 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.

[0024] The cosmetic composition can be manufactured into any dosage form that is normally manufactured in the industry. For example, it can be formulated into solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, surfactant-containing cleansers, oils, powder foundations, emulsion foundations, wax foundations, and sprays, etc., but is not limited thereto. For example, it can also be manufactured into dosage forms such as softening lotions, nutritive lotions, nutritive creams, massage creams, essences, eye creams, cleansing creams, cleansing foams, cleansing waters, packs, sprays, or powders.

[0025] When the dosage form of the cosmetic composition is a paste, cream, or gel, animal oils, vegetable oils, waxes, paraffins, starches, tragacanths, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicas, talcs, or zinc oxides, etc. can be used as the carrier component.

[0026] When the dosage form of the cosmetic composition is a powder or spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, or polyamide powder can be used as the carrier component. For example, in the case of a spray, it can further contain a propellant such as chlorofluorohydrocarbon, propane / butane, or dimethyl ether.

[0027] When the dosage form of the cosmetic composition is a solution or emulsion, a solvent, solubilizer, or emulsifier can be used as the carrier component. For example, it can contain water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyl glycol oil, glycerol fatty acid ester, polyethylene glycol, or sorbitan fatty acid ester.

[0028] When the dosage form of the cosmetic composition is a suspension, the carrier component may be a liquid diluent such as water, ethanol, or propylene glycol, a suspension agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, or polyoxyethylene sorbitan ester, or microcrystalline cellulose, aluminum methhydroxyl, bentonite, aga, or tracant.

[0029] When the dosage form of the cosmetic composition is a surfactant-containing cleanser, the carrier component may be an aliphatic alcohol sulfate, an aliphatic alcohol ether sulfate, a sulfosuccinate monoester, isethionate, imidazolinium derivative, methyl taurate, sarcosinate, fatty acid amide ether sulfate, alkylamide betaine, aliphatic alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, lanolin derivative, or ethoxylated glycerol fatty acid ester.

[0030] The peptide may be contained in nanosomes or nanoparticles to further improve skin penetration or stability issues. For example, the nanosomes may be produced by a microfluidizer using lecithin as a raw material and contained within lecithin particles. Any known method for producing the nanosomes may be used. 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, causing skin side effects, and if it exceeds 200 nm, skin penetration is not easy, making it difficult to obtain the benefits of using the nanosome structure.

[0031] In addition to the peptide and carrier components as active ingredients, the components contained in the cosmetic composition may include ingredients commonly used in cosmetic compositions, such as antioxidants, stabilizers, solubilizers, vitamins, pigments, and fragrances.

[0032] The content of peptides as active ingredients in the cosmetic composition can be appropriately and non-restrictively selected depending on the product form, desired use, etc., and can be added in amounts of, for example, 0.01 to 15% by weight of the total weight of the cosmetic composition. Alternatively, for example, the cosmetic composition may contain peptides in an amount of 1.0% to 3.0% by weight, preferably 2.0% to 3.0% by weight, based on the total weight.

[0033] Further embodiments include a method for improving skin condition, comprising the step of applying a cosmetic composition containing a peptide comprising the amino acid sequence of SEQ ID NO: 1 as an active ingredient to the skin of an individual; and providing uses for the peptide comprising the amino acid sequence of SEQ ID NO: 1 for the production of compositions for improving skin condition.

[0034] As stated above, any terms or elements mentioned in the description relating to the cosmetic composition that are the same as those already mentioned are as described above.

[0035] As used herein, the term “individual” means an individual subject requiring improvement of skin condition, and more specifically, a human or a mammal such as a non-human primate, mouse, dog, cat, horse, or cow.

[0036] As used herein, the terms “apply,” “administer,” and “coat” are interchangeable and may mean to bring about at least partial localization of the composition according to one embodiment to a desired site, or to place the composition according to one embodiment within an individual via an administration route.

[0037] Another embodiment provides an antioxidant composition containing a peptide consisting of the amino acid sequence of SEQ ID NO: 1 as an active ingredient.

[0038] As stated above, any terms or elements mentioned in the description of the peptides that are the same as those already mentioned are as described above.

[0039] The antioxidant composition may also be in the form of a pharmaceutical composition, a quasi-drug composition, or a cosmetic composition. For example, the composition may be used as a cosmetic composition for improving skin condition, or as a pharmaceutical composition for improving or treating the condition of diseases related to skin damage.

[0040] According to one embodiment, the peptide can restore the inhibited activity of fibroblasts and keratinocytes and enhance their antioxidant activity. Therefore, the peptide can be used as an active ingredient in antioxidant compositions.

[0041] The antioxidant composition may, for example, be provided in the form of a pharmaceutically effective composition. The pharmaceutically effective composition may also contain, but is not limited to, a pharmaceutically effective amount of the peptide; and / or a pharmaceutically acceptable carrier.

[0042] As used herein, the term "pharmaceutical effective amount" may mean an amount sufficient to achieve the preventive or therapeutic efficacy of the pharmaceutical composition for diseases related to skin damage.

[0043] The pharmaceutically acceptable carriers mentioned above are those commonly used in formulation and include, but are not limited to, lactose, dextrose, saccharose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methylcellulose, methylhydroxybenzoic acid, propylhydroxybenzoic acid, talc, magnesium stearate, and mineral oil. Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington's Pharmaceutical Sciences (19th ed., 1995).

[0044] The weight ratio between the peptide and the pharmaceutically acceptable carrier may be, for example, 500:1 to 1:500, and may also be, but is not limited to, 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.

[0045] The aforementioned pharmaceutical composition may further contain, but is not limited to, lubricants, wetting agents, sweeteners, flavoring agents, emulsifiers, suspension agents, preservatives, and the like, in addition to the aforementioned components.

[0046] The aforementioned pharmaceutical composition may be administered orally or parenterally, preferably parenterally, and 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 to these.

[0047] The dosage of the aforementioned 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 to these values ​​and can be prescribed in various ways depending on factors such as the formulation method, administration method, patient's age, weight, sex, medical condition, diet, administration time, route of administration, excretion rate, and response sensitivity.

[0048] The pharmaceutical composition may be manufactured in unit dose form or encapsulated in a multi-dose container by formulating it with pharmaceutically acceptable carriers and / or excipients using a method readily available to a person with ordinary skill in the art to which the invention pertains.

[0049] The dosage form may be in the form of a solution, suspension or emulsion in an oil or aqueous medium, or in the form of an extract, powder, granules, tablet or capsule, and may further contain a dispersant and / or stabilizer.

[0050] Another embodiment provides a food composition for improving skin condition that contains a peptide consisting of the amino acid sequence of SEQ ID NO: 1 as an active ingredient.

[0051] As stated above, any terms or elements mentioned in the description of the peptides that are the same as those already mentioned are as described above.

[0052] The content of the peptide as an active ingredient contained in the food composition can be appropriately and non-restrictively selected depending on the form of the food, the desired use, etc., and can be added in an amount of 0.01 to 15% by weight of the total food weight. Also, for example, in a health beverage composition, it can be added in a ratio of 0.02 to 10 g, preferably 0.3 to 1 g, based on 100 ml. [Effects of the Invention]

[0053] According to one embodiment of the peptide, it can be applied to improve skin conditions, including wrinkle reduction, improved skin elasticity, wound healing, strengthening of the skin barrier, or suppression of skin aging, by promoting the proliferation of fibroblasts and keratinocytes and increasing the synthesis of extracellular matrix components and skin barrier factors.

[0054] According to one embodiment of the peptide, it may help restore the inhibited activity of fibroblasts and keratinocytes and enhance their antioxidant activity, thereby contributing to the recovery of skin damage caused by external environmental factors such as ultraviolet radiation.

[0055] Therefore, a peptide according to one embodiment can be used as an active ingredient in a composition for improving skin condition or an antioxidant composition. [Brief explanation of the drawing]

[0056] [Figure 1]This is the result of adding a peptide consisting of the amino acid sequence of SEQ ID NO: 1 to NIH3T3 cells and confirming the level of cell proliferation by measuring changes in cell viability. [Figure 2] This study shows the results of adding a peptide consisting of the amino acid sequence of Sequence ID No. 1 to NIH3T3 cells, after which increased expression of the extracellular matrix components Col1a1, fibronectin, and elastin was confirmed. [Figure 3] The expression levels of extracellular matrix components were quantitatively evaluated after adding a peptide consisting of the amino acid sequence of SEQ ID NO: 1 to NIH3T3 cells. Figure 3A shows the results of confirming the expression level of Col1a1, Figure 3B shows the results of confirming the expression level of fibronectin, and Figure 3C shows the results of confirming the expression level of elastin. [Figure 4] This result shows that after adding a peptide consisting of the amino acid sequence of SEQ ID NO: 1 to HaCaT cells, an increase in the expression of SIRT-1 and AQP3, which are skin barrier factors, was confirmed. [Figure 5] The expression levels of skin barrier factors were quantitatively evaluated after adding a peptide consisting of the amino acid sequence of SEQ ID NO: 1 to HaCaT cells. Figure 5A shows the results of confirming the expression level of SIRT-1, and Figure 5B shows the results of confirming the expression level of AQP3. [Figure 6] This study confirmed the inhibitory effect of a peptide consisting of the amino acid sequence of SEQ ID NO: 1 on UV-induced skin cell death after adding it to NIH3T3 cells, through survival rate evaluation. [Figure 7] This study confirmed the inhibitory effect of adding a peptide consisting of the amino acid sequence of SEQ ID NO: 1 to HaCaT cells, and then evaluating the survival rate to suppress the death of skin cells induced by UV irradiation. [Figure 8] This is the result of quantitatively confirming the change in reactive oxygen species levels, which were increased by ultraviolet light, after adding a peptide consisting of the amino acid sequence of SEQ ID NO: 1 to NIH3T3 cells. [Figure 9]This is the result of quantitatively confirming the change in reactive oxygen species levels, which were increased by ultraviolet light, after adding a peptide consisting of the amino acid sequence of SEQ ID NO: 1 to HaCaT cells. [Modes for carrying out the invention]

[0057] The present invention will be described in more detail below based on examples. However, these examples are for illustrative purposes only, and the scope of the present invention is not limited to these examples.

[0058] Example 1. Synthesis of peptides Peptides having the amino acid sequence of SEQ ID NO: 1, as shown in [Table 1] below, were synthesized using an automated peptide synthesizer (Milligen 9050, Millipore, USA), and these synthesized peptides were separated into pure molecules using C18 reversed-phase high-performance liquid chromatography (HPLC) (Waters Associates, USA). The column used was ACQUITY UPLC BEH300 C18 (2.1 mm x 100 mm, 1.7 μm, Waters Co, USA).

[0059] [Table 1]

[0060] Example 2. Confirmation of the proliferation-promoting effect on skin cells In this example, the effect of the peptide from this example on skin cell proliferation was confirmed by evaluating the change in the viability of NIH3T3 cells, which are mouse fibroblasts.

[0061] Specifically, 1 x 10⁶ NIH3T3 cells in a 96-well plate 4After seeding at a density of cells / well, the cells were cultured for 24 hours. Subsequently, the cells were washed once with serum-free DMEM media, and 50 μM or 100 μM of the peptide consisting of the amino acid sequence of SEQ ID NO: 1 was dispensed into 200 μL of the serum-free media. This was then cultured in a CO2 incubator at 37°C for 72 hours. Subsequently, the culture was washed twice with PBS, and then MTT solution was dispensed into each well at a concentration of 0.5 mg / ml. After shielding from light, the culture was cultured in a CO2 incubator at 37°C for 4 hours, and the absorbance was measured at 540 nm using a microplate reader (Molecular Devices, USA).

[0062] As a result, as shown in Figure 1, we confirmed that the peptide consisting of the amino acid sequence of Sequence ID No. 1 promotes the proliferation of fibroblasts.

[0063] Example 3. Confirmation of wrinkle improvement and elasticity enhancement effects. In this example, the effects of the peptide from this example on improving intrinsic skin aging, including wrinkle reduction and increased elasticity, were confirmed by evaluating the expression levels of collagen type 1 (Col1a1), fibronectin, and elastin, which are known components of the dermis.

[0064] Specifically, NIH3T3 cells were placed in a 6-well plate in a 3 x 10⁶ arrangement. 5After seeding at a density of cells / well, the cells were cultured for 24 hours. Subsequently, the cells were washed once with serum-free DMEM media, and then 10 μM or 50 μM of the peptide consisting of the amino acid sequence of SEQ ID NO: 1 was dispensed into 1 mL of the culture medium and cultured in a CO2 incubator at 37°C for 24 hours. Subsequently, the culture was washed twice with PBS, and RNA was isolated from the culture using easy blue (iNtRON, Cat. No.: 17061, Korea). Subsequently, the isolated RNA was reverse transcribed using an RT kit (Enzynomics, Cat. No.: RT200, Korea) to synthesize cDNA. Subsequently, polymerase chain reaction (PCR) was performed using the synthesized 1 μg cDNA, primers for Col1a1, fibronectin, or elastin, and a PCR kit (enzynomics, Cat. No.: P581T, Korea). On the other hand, in this example, the control group was the untreated group, and the positive control group was the group to which TGF-β1 was added. The nucleotide sequences of the primers used in this example are shown in Table 2 below.

[0065] [Table 2]

[0066] As a result, as shown in Figures 2 and 3, we confirmed that the peptide consisting of the amino acid sequence of Sequence ID No. 1 increased the expression of the extracellular matrix components Col1a1, fibronectin, and elastin.

[0067] Example 4. Confirmation of skin barrier strengthening effect In this example, we attempted to confirm the effect of the peptide described in this example on strengthening the skin barrier by evaluating the expression levels of sirtuin-1 (SIRT-1) or aquaforin-3 (AQP3).

[0068] Specifically, HaCaT cells were placed in a 6-well plate in a 3 x 10⁶ arrangement. 5 After seeding at a density of cells / well, the cells were cultured for 24 hours. Subsequently, the cells were washed once with serum-free DMEM media, and then 50 μM or 100 μM of the peptide consisting of the amino acid sequence of SEQ ID NO: 1 was dispensed into 1 mL of the culture medium and cultured in a CO2 incubator at 37°C for 24 hours. Subsequently, the culture was washed twice with PBS, and RNA was isolated from the culture using easy blue (iNtRON, Cat. No.: 17061, Korea). Subsequently, the isolated RNA was reverse transcribed using an RT kit (Enzynomics, Cat. No.: RT200, Korea) to synthesize cDNA. Subsequently, a polymerase chain reaction (PCR) was performed using the synthesized 1 μg cDNA, primers for SIRT-1 or AQP3, and a PCR kit (enzynomics, Cat. No.: P581T, Korea). On the other hand, in this embodiment, the control group was the untreated group, and the positive control group was the group to which EGF was added. The nucleotide sequences of the primers used in this embodiment are shown in Table 3 below.

[0069] [Table 3]

[0070] As a result, as shown in Figures 4 and 5, it was confirmed that the peptide consisting of the amino acid sequence of Sequence ID No. 1 increased the expression of SIRT-1 and AQP3, which are skin barrier factors. Through these results, it was found that the peptide according to one example contributes to strengthening the skin barrier and anti-aging of the skin by increasing skin barrier factors.

[0071] Example 5. Confirmation of the inhibitory effect on UV-induced skin cell death. In this example, we evaluated the change in the survival rate of skin cells whose death was induced by ultraviolet irradiation, thereby confirming the effect of the peptide according to this example on suppressing the death of skin cells induced by ultraviolet radiation or on cell recovery.

[0072] Specifically, 1 × 10⁶ NIH3T3 cells or HaCaT cells are placed in a 96-well plate. 4 After seeding at a density of cells / well, the cells were cultured for 24 hours. Subsequently, the cells were washed once with serum-free DMEM media, and 50 μM or 100 μM of the peptide consisting of the amino acid sequence of SEQ ID NO: 1 was dispensed into 200 μL of the culture medium. The culture medium containing the peptide was incubated in a CO2 incubator at 37°C for 1 hour, then transferred to an e-tube, mixed with 100 μL of PBS, and dispensed into well plates. Subsequently, UV irradiation (VILBER LOURMAT, Cat No.: 3102-BSU, France) was used to target NIH3T3 cells at 6 J / cm². 2 Ultraviolet light was irradiated, and HaCaT cells were targeted with 15 mJ / cm² of ultraviolet light. 2 The cultures were irradiated with ultraviolet light. Subsequently, the PBS in the well plate was removed, and MTT (Sigma, Cat. No.: M2003, USA) solution was dispensed into each well at a concentration of 0.5 mg / ml. The cultures were then incubated in a CO2 incubator at 37°C for 72 hours. The absorbance of the cultures at 540 nm was then measured using a microplate reader (Molecular Devices, USA). In this example, the control group was the untreated group, and the positive control group was the group treated with Trolox (Trolox).

[0073] As a result, as shown in Figures 6 and 7, it was confirmed that the peptide consisting of the amino acid sequence of Sequence ID No. 1 restored the cell viability of fibroblasts and keratinocytes that had decreased due to UV irradiation. Through these results, it was found that the peptide according to one example contributes to suppressing UV-induced skin cell death.

[0074] Example 6. Confirmation of the effect of reducing reactive oxygen species increased by ultraviolet rays In this example, by evaluating the change in the level of reactive oxygen species in skin cells increased by ultraviolet irradiation, the effect of the peptide according to one example on the antioxidant effect in damaged skin cells was confirmed.

[0075] Specifically, NIH3T3 cells or HaCaT cells were seeded in a 6-well plate at a density of 5×10 5 cells / well and then cultured for 24 hours. Thereafter, the cells were washed once with serum-free DMEM media, and then the peptide consisting of the amino acid sequence of SEQ ID NO: 1 was dispensed at 50 μM or 100 μM per 1 mL of the medium, and this was cultured in a CO2 incubator at 37 °C for 1 hour. Thereafter, the culture was transferred to an e-tube, 1 ml of PBS was mixed and dispensed into the well plate. Thereafter, using a UV irradiation machine (VILBER LOURMAT, Cat No.: 3102-BSU, France), NIH3T3 cells were irradiated with ultraviolet rays of 6 J / cm 2 and HaCaT cells were irradiated with ultraviolet rays of 15 J / cm 2 . Thereafter, after removing the PBS in the well plate, 900 μL of the culture medium into which the peptide was dispensed was added, and this was cultured in a CO2 incubator at 37 °C for 24 hours. Thereafter, 10 μM of DCFG-DA (2’,7’-dichlorofluorescin diacetate) was treated to the culture, wrapped with foil, and then this was cultured in a CO2 incubator at 37 °C for 30 minutes. Thereafter, this was washed twice with PBS, 500 μL of 1× trypsin / EDTA was treated here to obtain cells, and then centrifugation was performed. After washing the centrifuged cells with PBS, the FL1 fluorescence value was measured through flow cytometry (FACS, BD, USA). In this example, the control group was the non-treated group, the comparison group was the group irradiated only with ultraviolet rays, and as the positive control group, the group to which Trolox was added (Trolox) was used.

[0076] As a result, as shown in Figures 8 and 9, it was confirmed that the peptide consisting of the amino acid sequence of Sequence ID No. 1 reduced the levels of reactive oxygen species in fibroblasts and keratinocytes that were increased by UV irradiation. Through these results, it was found that the peptide according to one example contributes to the antioxidant effect on UV-induced skin cells.

[0077] Dosage Form Example 1: Production of Peptide Nanosomes 50 mg of the peptide from Example 1 was dissolved in 500 ml of distilled water by thorough stirring. The mixture was then mixed with 5 g of lecithin, 0.3 ml of sodium oleate, 50 ml of ethanol, and a small amount of oil. After adjusting the volume with distilled water to a total of 1 L, the mixture was emulsified using a microfluidizer under high pressure to produce peptide nanosomes with a size of approximately 100 nm.

[0078] Dosage form example 2. Softening lotion A softening lotion containing a peptide according to one embodiment and having the following composition was manufactured using a method known in the industry.

[0079] [Table 4]

[0080] Dosage form example 3: Nutritional cream A nutritional cream containing a peptide according to one embodiment and having the following composition was manufactured using a method known in the industry.

[0081] [Table 5]

[0082] Dosage form example 4. Nourishing lotion A nourishing lotion containing a peptide according to one embodiment and having the following composition was manufactured using a method known in the industry.

[0083] [Table 6]

[0084] Dosage Form Example 5: Essence An essence containing a peptide according to one embodiment and having the following composition was prepared using a method known in the industry.

[0085] [Table 7]

[0086] The above description of the present invention is illustrative, and a person with ordinary skill in the art to which the invention pertains will understand that it can be easily modified into other specific forms without altering the technical idea or essential features of the invention. Therefore, the embodiments described above should be understood to be illustrative in all respects and not limiting.

Claims

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

1.

2. The N-terminus of the aforementioned peptide is an acetyl group, fluorenyl methoxycarbon Fluoroyl group (fluoreonylmethoxycarbonyl group), formyl group, palmitoyl group ( palmitoyl group, myristyl group, stearyl group, b Toxycarbonyl group (butoxycarbonyl group), allyloxycarbonyl group (allyloxycarbonyl group) Selected from the group consisting of nyl group and polyethylene glycol (PEG). The peptide according to claim 1, which is bonded to any one of the protecting groups.

3. The C-terminus of the peptide is an amino group (-NH₂). 2 ), tertiary alkyl group (tertiary a lkyl group) and azide (-NHNH 2 One protection selected from the group consisting of ) The peptide according to claim 1, which is bonded to a group.

4. Claim 1, the peptide exhibits one or more of the following characteristics. Peptides listed: (a) Promoting the proliferation of fibroblasts or keratinocytes; (b) Extracellular matrix components such as collagen type 1 (Col1a1), fibronectin, and This involves increased elastin expression; (c) Sirtuin-1 (SIRT-1) or aquaforin-3 (AQP), which are skin barrier factors. 3) Increased expression; and (d) Inhibition of fibroblast or keratinocyte death and reduction of reactive oxygen species levels.

5. A skin-like substance comprising the peptide described in any one of claims 1 to 4 as an active ingredient. A cosmetic composition for improving appearance.

6. The peptide is formulated into nanosomes, according to claim 5. A compositional material.

7. The aforementioned improvement in skin condition includes wrinkle reduction, improved skin elasticity, wound healing, strengthening of the skin barrier, or skin The cosmetic composition according to claim 5, which inhibits aging.

8. The cosmetic composition according to claim 7, wherein the skin aging is skin aging caused by ultraviolet light.

Images