Peptide having skin condition improving activity and use thereof

Abstract

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

Description

[Technical field]

[0001] The present invention relates to a peptide having skin condition-improving activity and uses thereof. [Background technology]

[0002] Human skin undergoes constant changes, the most typical of which are the decline in skin function and visual beauty due to aging. Aging causes wrinkles in the skin, and typical factors that cause wrinkles include exposure to ultraviolet light and reduced collagen synthesis. Skin aging is broadly divided into intrinsic aging caused by genetic factors and extrinsic aging caused by external environmental factors such as sunlight. Among these, it is known that extrinsic aging can be prevented, treated, or delayed through the removal of active oxygen, the proliferation of fibroblasts, and the promotion of collagen synthesis.

[0003] Collagen, a major component of the extracellular matrix, is the main matrix protein produced by skin fibroblasts. Collagen forms most of the organic matter in skin, tendons, bones, and teeth, but is particularly abundant in bones and skin (dermis). Such collagen is reduced by age and photoaging caused by exposure to ultraviolet light, 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 and allowing wounds to heal quickly and without scars. It has also been reported that if the density of the basal layer etc. is increased by promoting collagen biosynthesis, the melanin pigment concentration per unit skin density will be reduced, and the effect of brightening the skin tone can be expected.

[0004] Against this technical background, multifaceted research is being conducted to improve skin conditions through mechanisms such as promoting collagen biosynthesis and increasing the proliferation and activity of fibroblasts and other cells (Patent Document 1), but the reality is that there are still shortcomings. [Prior art documents] [Patent documents]

[0005] [Patent Document 1] Korean Patent No. 10-1813629 Summary of the Invention [Problem to be solved by the invention]

[0006] One embodiment provides a peptide comprising the amino acid sequence of SEQ ID NO:1.

[0007] Another aspect provides a cosmetic composition for improving skin conditions, which comprises a peptide having the amino acid sequence of SEQ ID NO:1 as an active ingredient.

[0008] Other objects and advantages of the present application will become more apparent from the following detailed description, together with the claims and drawings. Contents not described in this specification can be fully recognized and inferred by those skilled in the art of the present application or a similar art, so the description thereof will be omitted. [Means for solving the problem]

[0009] Each description and embodiment disclosed in this application may be applied to each other description and embodiment. That is, all combinations of various elements disclosed in this application belong to the scope of this application. In addition, the scope of this application cannot be limited by the specific descriptions described below.

[0010] One embodiment provides a peptide consisting of the amino acid sequence of SEQ ID NO:1.

[0011] The term "peptide" as used herein may refer to a linear molecule formed by binding amino acid residues to each other by peptide bonds. The peptide may be prepared by chemical synthesis methods known in the art, particularly solid-phase synthesis or liquid synthesis (U.S. Patent No. 5,516,891). The present inventors have made extensive efforts to develop a peptide having biologically effective activity, and have identified a peptide having the amino acid sequence of SEQ ID NO: 1. Here, the biologically effective activity may be any one or more of the following characteristics: (a) promotion of proliferation of fibroblasts or keratinocytes; (b) enhancement of expression of type I collagen (Col1a1), fibronectin, or elastin, which are components of the extracellular matrix; (c) enhancement of expression of sirtuin 1 (SIRT-1) or aquaporin 3 (AQP3), which are skin barrier factors; and (d) inhibition of death of fibroblasts or keratinocytes, and reduction of reactive oxygen levels. Therefore, the peptide can be used for improving skin conditions or as an antioxidant.

[0012] The peptides may also be conjugated with protecting groups at their N- or C-termini to obtain chemical stability, enhanced pharmacological properties (half-life, absorbency, potency, efficacy, etc.), altered specificity (e.g., broader spectrum of biological activity), and reduced antigenicity. In one embodiment, the N-terminus of the peptide may be bound to any one of the protecting groups selected from the group consisting of acetyl group, fluorenylmethoxycarbonyl group, formyl group, palmitoyl group, myristyl group, stearyl group, butoxycarbonyl group, allyloxycarbonyl group, and polyethylene glycol (PEG), and / or the C-terminus of the peptide may be bound to any one of the protecting groups selected from the group consisting of amino group (-NH2), tertiary alkyl group, and azide (-NHNH2). The peptide may also optionally further comprise a targeting sequence, a tag, a labeled residue, or a specifically engineered amino acid sequence to increase half-life or peptide stability.

[0013] The term "stability" as used herein may refer not only to in vivo stability, which protects the peptide from attack by in vivo proteolytic enzymes, but also to storage stability (eg, room temperature storage stability).

[0014] Another aspect provides a cosmetic composition for improving skin conditions, which comprises a peptide having the amino acid sequence of SEQ ID NO:1 as an active ingredient.

[0015] Among the terms or elements referred to in the description of the peptide, those that are the same as those already mentioned are as described above.

[0016] The term "amelioration" as used herein may refer to any action that at least reduces the parameters associated with the alleviation or treatment of a condition, for example, the severity of a symptom.

[0017] The term "improving skin condition" as used herein comprehensively means the process or effect of treating, reducing or alleviating skin damage induced by intrinsic or extrinsic factors, and may be interpreted as meaning, for example, improvement of wrinkles, improvement of skin elasticity, wound recovery, strengthening of the skin barrier or inhibition of skin aging, but is not limited thereto.

[0018] Here, "wrinkle improvement," "skin elasticity improvement," and "wound recovery" may refer to any action of increasing the total amount of extracellular matrix factors, including promoting collagen synthesis. "Strengthening the skin barrier" may refer to strengthening the skin's natural function of preventing moisture and nutrients from leaking out of the skin and preventing harmful substances such as bacteria and viruses from penetrating the skin. "Skin aging inhibition" may refer to inhibition of deterioration of skin functions such as wrinkles, sagging skin, and reduced elasticity. Here, the skin aging may also be photoaging, for example, skin aging caused by ultraviolet rays.

[0019] Conventional functional peptides, despite their effective biological activity, have the disadvantages that they cannot be effectively delivered to target tissues or cells due to the size of the peptide itself, or that they disappear in the body in a short period of time due to their short half-life. In addition, the cosmetic composition according to one embodiment contains a peptide consisting of 10 or less amino acids as an active ingredient, which has excellent skin penetration rate of the active ingredient, and for example, when applied topically to the skin, it can effectively improve the skin condition.

[0020] According to one embodiment, it was shown that the peptide can promote the proliferation of fibroblasts and keratinocytes, and enhance the synthesis of extracellular matrix components and skin barrier factors. It was also 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 a cosmetic composition for improving skin condition.

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

[0022] The term "cosmetically effective amount" as used herein means an amount sufficient to achieve the skin condition improving efficacy of the cosmetic composition.

[0023] The weight ratio of the peptide to the cosmetically acceptable carrier may be, for example, 500:1 to 1:500. By way of example, the weight ratio may 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 may be prepared in any formulation commonly used in the art, such as, but not limited to, a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleanser, oil, powder foundation, emulsion foundation, wax foundation, spray, etc. For example, it may be prepared in the form of a softening lotion, nourishing lotion, nourishing cream, massage cream, essence, eye cream, cleansing cream, cleansing foam, cleansing water, pack, spray, or powder.

[0025] When the cosmetic composition is in the form of a paste, cream or gel, the carrier component may be an animal oil, a vegetable oil, wax, paraffin, starch, tragacanth, a cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide.

[0026] When the cosmetic composition is in the form of a powder or spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, or polyamide powder is used as a carrier component, and when the cosmetic composition is in the form of a spray, for example, a propellant such as chlorofluorohydrocarbon, propane / butane, or dimethyl ether may be additionally included.

[0027] When the cosmetic composition is in the form of a solution or emulsion, a solvent, solubilizer, or emulsifier 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 fatty acid esters, polyethylene glycol, or fatty acid esters of sorbitan.

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

[0029] When the cosmetic composition is in the form of a surfactant-containing cleanser, fatty alcohol sulfates, fatty alcohol ether sulfates, sulfosuccinic acid monoesters, isethionates, imidazolinium derivatives, methyl taurate, sarcosinates, fatty acid amide ether sulfates, alkylamido betaines, fatty alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives, or ethoxylated glycerol fatty acid esters may be used as carrier components.

[0030] The peptide may be contained in nanosomes or nanoparticles to further improve skin permeation or stability problems. For example, the nanosomes may be prepared by a microfluidizer using lecithin as a raw material, and may be contained in lecithin particles. Any known method may be used to prepare the nanosomes. The nanosome particles preferably have a size of 30 to 200 nm. If the nanosome particles have a size of less than 30 nm, they may penetrate the skin too quickly and cause skin side effects, and if the size exceeds 200 nm, they may not penetrate the skin easily, making it difficult to obtain the effect of using the nanosome structure.

[0031] The components contained in the cosmetic composition include, in addition to the peptide as an active ingredient and the carrier component, components generally used in cosmetic compositions, and may also include general adjuvants such as antioxidants, stabilizers, solubilizers, vitamins, pigments, and fragrances.

[0032] The content of the peptide as an active ingredient contained in the cosmetic composition is appropriately selected without limitation depending on the product form, desired use, etc., and may be added in an amount of, for example, 0.01 to 15 wt % of the total cosmetic composition weight. Also, for example, the cosmetic composition may contain the peptide in an amount of 1.0 to 3.0 wt %, preferably 2.0 to 3.0 wt %, based on the total weight.

[0033] Yet another aspect provides a method for improving skin condition, comprising the step of applying to the skin of an individual a cosmetic composition comprising as an active ingredient a peptide having the amino acid sequence of SEQ ID NO:1, and use of the peptide having the amino acid sequence of SEQ ID NO:1 for the manufacture of a composition for improving skin condition.

[0034] Among the terms and elements referred to in the description of the cosmetic composition, those that are the same as those already mentioned are as described above.

[0035] As used herein, the term "individual" refers to a subject in need of improved skin condition, and more specifically refers to a mammal, such as a human or non-human primate, mouse, dog, cat, horse, or cow.

[0036] As used herein, the terms "apply," "administer," and "apply" are used interchangeably and may refer to causing at least partial localization of a composition according to an embodiment to a desired site, or disposing a composition according to an embodiment within an individual by a route of administration.

[0037] Yet another embodiment provides an antioxidant composition comprising a peptide having the amino acid sequence of SEQ ID NO:1 as an active ingredient.

[0038] Among the terms or elements referred to in the description of the peptide, those that are the same as those already mentioned are as described above.

[0039] The antioxidant composition may be in the form of a pharmaceutical composition, a quasi-drug composition, or a cosmetic composition. For example, the composition may be used in a cosmetic composition for improving skin conditions, or a pharmaceutical composition for improving or treating the condition of a disease associated with skin damage.

[0040] According to one embodiment, the peptide was shown to be capable of restoring the inhibited activity of fibroblasts and keratinocytes and enhancing their antioxidant activity, and therefore, the peptide can be used as an active ingredient in an antioxidant composition.

[0041] The antioxidant composition may be provided in the form of a pharmaceutical composition, for example, which may include, but is not limited to, a pharma- ceutical effective amount of the peptide and / or a pharma- ceutical acceptable carrier.

[0042] The term "pharmaceutical effective amount" as used herein may refer to an amount sufficient to achieve preventive or therapeutic efficacy of the pharmaceutical composition against a disease associated with skin damage.

[0043] The pharma- ceutically acceptable carriers are those commonly used in formulations, and include, but are not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, starch, gum acacia, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methylcellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil, etc. Suitable pharma-ceutically acceptable carriers and formulations are described in detail in Remington's Pharmaceutical Sciences (19th ed., 1995).

[0044] The weight ratio of said peptide to said pharma- ceutically acceptable carrier may be, for example, 500:1 to 1:500. By way of example, and not by way of limitation, 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 2:1 to 1:2.

[0045] The pharmaceutical composition may further contain, in addition to the above components, lubricants, wetting agents, sweeteners, flavoring agents, emulsifiers, suspending agents, preservatives, etc., but is not limited thereto.

[0046] The 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, peritoneal injection, topical administration, transdermal administration, etc., but is not limited thereto.

[0047] The dosage of the pharmaceutical composition may be, but is not limited to, 0.0001 to 1,000 μg, 0.001 to 1,000 μg, 0.01 to 1,000 μg, 0.1 to 1,000 μg, or 1.0 to 1,000 μg per day, and may be variously prescribed depending on factors such as formulation method, administration method, age, weight, sex, pathological condition of the patient, food, administration time, administration route, excretion rate, and reaction sensitivity.

[0048] The pharmaceutical composition may be prepared in a unit dose form or in a multi-dose container by formulating the composition using pharma- ceutically acceptable carriers and / or excipients in a manner that can be easily carried out by a person having ordinary skill in the art to which the invention pertains.

[0049] The dosage forms may be in the form of a solution, suspension or emulsion in an oily or aqueous medium, or in the form of an extract, powder, granules, tablets or capsules, which may additionally contain dispersing agents and / or stabilizers.

[0050] Yet another embodiment provides a food composition for improving skin conditions, comprising a peptide having the amino acid sequence of SEQ ID NO:1 as an active ingredient.

[0051] Among the terms or elements referred to in the description of the peptide, those 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 is appropriately selected without limitation depending on the type of food, the desired use, etc., and may be added at, for example, 0.01 to 15% by weight of the total food weight. Also, for example, the peptide may be added at a ratio of 0.02 to 10 g, preferably 0.3 to 1 g, based on 100 ml of the health drink composition. Effect of the Invention

[0053] According to one embodiment of the peptide, the proliferation of fibroblasts and keratinocytes is promoted, and synthesis of components of the extracellular matrix and skin barrier factors is increased, whereby the peptide can be applied to improve skin conditions, including wrinkle improvement, improvement of skin elasticity, wound recovery, strengthening of the skin barrier, and inhibition of skin aging.

[0054] According to one embodiment, the peptide can contribute to repairing skin damage that may be caused by external environment such as ultraviolet rays by restoring the inhibited activity of fibroblasts and keratinocytes and enhancing their antioxidant efficacy.

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

[0056] [Figure 1] The peptide having the amino acid sequence of SEQ ID NO: 1 was added to skin cells, and then the cell proliferation level was confirmed. (A) shows the change in viability of NIH3T3 cells, and (B) shows the change in viability of HaCaT cells. [Diagram 2] This is the result of confirming an increase in the expression of Col1a1, fibronectin and elastin, which are components of the extracellular matrix, after adding a peptide having the amino acid sequence of SEQ ID NO: 1 to NIH3T3 cells. [Diagram 3] A peptide having the amino acid sequence of SEQ ID NO: 1 was added to NIH3T3 cells, and the expression levels of extracellular matrix components were quantitatively evaluated. A is the result of confirming the expression level of Col1a1, B is the result of confirming the expression level of fibronectin, and C is the result of confirming the expression level of elastin. [Figure 4] This is a result of confirming increased expression of skin barrier factors SIRT-1 and AQP3 after adding a peptide having the amino acid sequence of SEQ ID NO: 1 to HaCaT cells. [Diagram 5] A peptide having the amino acid sequence of SEQ ID NO: 1 was added to HaCaT cells, and then the expression level of skin barrier factors was quantitatively evaluated. A shows the results of confirming the expression level of SIRT-1, and B shows the results of confirming the expression level of AQP3. [Figure 6] After adding a peptide having the amino acid sequence of SEQ ID NO: 1 to skin cells, the inhibitory effect on the death of skin cells induced by ultraviolet light irradiation was confirmed through viability evaluation. A shows the results showing the change in viability of NIH3T3 cells, and B shows the results showing the change in viability of HaCaT cells. [Figure 7] After adding a peptide having the amino acid sequence of SEQ ID NO: 1 to skin cells, the antioxidant effect on the skin cells induced by ultraviolet light irradiation was confirmed through an evaluation of the reactive oxygen level. A shows the change in the reactive oxygen level of NIH3T3 cells, and B shows the change in the reactive oxygen level of HaCaT cells. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

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

[0058] Example 1. Synthesis of peptides Using an automated peptide synthesis group (Milligen 9050 (Millipore, USA)), peptides having the amino acid sequence of SEQ ID NO: 1 shown in Table 1 below were synthesized, and the synthesized peptides were purified and separated using C18 reversed-phase high performance liquid chromatography (HPLC) (Waters Associates, USA). The column used was an ACQUITY UPLC BEH300 C18 (2.1 mmX100 mm, 1.7 μm (Waters Co., USA).

[0059] [Table 1]

[0060] Example 2. Confirmation of the effect of promoting proliferation of skin cells In this example, the effect of a peptide according to one embodiment on the proliferation of skin cells is confirmed by evaluating changes in the viability of NIH3T3 cells, which are mouse fibroblasts, or HaCaT cells, which are human keratinocytes.

[0061] Specifically, NIH3T3 cells or HaCaT cells were cultured in a 96-well plate at 5 × 10 3After seeding at a density of cells / well, the cells were cultured for 24 hours. The cells were then washed once with serum-free DMEM media, and the culture medium was replaced with the serum-free medium. Then, 50 μM or 100 μM of peptides consisting of the amino acid sequence of SEQ ID NO: 1 were dispensed into the medium, and the medium was cultured in a CO2 incubator at 37°C for 72 hours. The culture was then washed twice with PBS, and treated with 4% paraformaldehyde (PFA) solution for 15 minutes to fix the cultured cells. Then, the culture was washed twice with distilled water, treated with SRB solution for 1 hour, cell stained, washed with 1% acetic acid solution, and dried. The dried culture was then dissolved in 20 mM Tris buffer, and the absorbance at 560 nm was measured using a microplate reader (Molecular Devices, USA). In this example, a non-treated group was used as a control group, and a group to which insulin-like growth factor (IGF) was added was used as a positive control group.

[0062] As a result, as shown in FIG. 1, it was found that the peptide consisting of the amino acid sequence of SEQ ID NO: 1 promoted the proliferation of fibroblasts and keratinocytes.

[0063] Example 3. Confirmation of wrinkle improvement and elasticity improvement effects In this example, the expression levels of type I collagen (Col1a1), fibronectin, and elastin, which are known to be components of the dermis, are evaluated to confirm the effect of a peptide according to one embodiment on improving intrinsic skin aging, including improving wrinkles and increasing elasticity.

[0064] Specifically, NIH3T3 cells were cultured in a 6-well plate at 3 × 10 5After seeding at a density of cells / well, the cells were cultured for 24 hours. The cells were then washed once with serum-free DMEM media, and the culture medium was replaced with the serum-free medium. Then, 50 μM or 100 μM of a peptide having the amino acid sequence of SEQ ID NO: 1 was dispensed into the medium, and the medium was cultured for 24 hours in a CO2 incubator at 37°C. The culture was then washed twice with PBS, and RNA was isolated from the culture using easy BLUE (iNtRON, Korea). 1 μg of the isolated RNA was reverse transcribed using an RT Kit (Enzynomics, Korea) to synthesize each cDNA. Then, a polymerase chain reaction (PCR) was performed using the cDNA and primers for Col1a1, fibronectin, and elastin. The control group was an untreated group, and the positive control group was a group to which TGF-β1 was added (TGF-β1). The nucleotide sequences of the primers used in this example are as shown in Table 2 below.

[0065] [Table 2]

[0066] As a result, as shown in Figures 2 and 3, it was found that the expression of Col1a1, fibronectin and elastin, which are components of the extracellular matrix, was increased by the peptide having the amino acid sequence of SEQ ID NO: 1. From the above results, it was found that the peptide according to one embodiment contributes to the improvement of intrinsic skin aging, including wrinkle improvement and elasticity improvement, by increasing the components of the extracellular matrix.

[0067] Example 4. Confirmation of the effect of strengthening the skin barrier In this example, the effect of a peptide according to one embodiment on strengthening the skin barrier is confirmed by evaluating the expression levels of sirtuin 1 (SIRT-1) or aquaporin 3 (AQP3).

[0068] Specifically, HaCaT cells were cultured in a 6-well plate at 3 × 10 5 After seeding at a density of cells / well, the cells were cultured for 24 hours. The cells were then washed once with serum-free DMEM media, and the culture medium was replaced with the serum-free medium. Then, 50 μM or 100 μM of a peptide having the amino acid sequence of SEQ ID NO: 1 was dispensed into the medium, and the medium was cultured in a CO2 incubator at 37°C for 24 hours. The culture was then washed twice with PBS, and RNA was isolated from the culture using easy BLUE (iNtRON, Korea). 1 μg of the isolated RNA was reverse transcribed using an RT Kit (Enzynomics, Korea) to synthesize each cDNA. Then, polymerase chain reaction was performed using the cDNA and primers for sirtuin 1 and aquaporin 3. The control group was a non-treated group, and the positive control group was a group to which EGF was added (EGF). The nucleotide sequences of the primers used in this example are shown in Table 3 below.

[0069] [Table 3]

[0070] As a result, as shown in Figures 4 and 5, it was found that the expression of skin barrier factors SIRT-1 and AQP3 was increased by the peptide having the amino acid sequence of SEQ ID NO: 1. From the above results, it was found that the peptide according to one embodiment contributes to strengthening the skin barrier and anti-aging of the skin by increasing the skin barrier factor.

[0071] Example 5. Confirmation of the inhibitory effect on UV-induced skin cell death In this example, the effect of a peptide according to one embodiment on the inhibition of UV-induced death of skin cells or cell recovery is confirmed by evaluating the change in viability of skin cells whose death was induced by UV irradiation.

[0072] Specifically, NIH3T3 cells or HaCaT cells were cultured in a 96-well plate at 1 × 10 4 After seeding at a density of cells / well, the cells were cultured for 24 hours. The cells were then washed once with serum-free DMEM media, and the culture medium was replaced with the serum-free medium, to which the peptide having the amino acid sequence of SEQ ID NO: 1 was dispensed at 50 μM or 100 μM. The culture medium to which the peptide was dispensed was cultured in a CO2 incubator at 37°C for 1 hour, transferred to an E-TUBE, mixed with 100 μl of PBS, and dispensed into a well plate. Then, NIH3T3 cells were exposed to 6 J / cm irradiated using an ultraviolet irradiator (Vilber Lourmat, France). 2 HaCaT cells were exposed to 15 mJ / cm of UV light. 2 The well plate was then irradiated with ultraviolet light of 1000 nm. The PBS in the well plate was then removed, and 900 μL of the culture medium containing the peptide was added, followed by culturing for 72 hours in a CO2 incubator at 37°C. The culture was then washed twice with PBS, and treated with 4% paraformaldehyde (PFA) solution for 15 minutes to fix the cultured cells. The culture was then washed twice with distilled water, treated with SRB solution for 1 hour, stained, washed with 1% acetic acid solution, and dried. The dried culture was then dissolved in 20 mM Tris buffer, and the absorbance at 560 nm was measured using a microplate reader (Molecular Devices, USA). In this example, a non-treated group was used as the control group, and a group containing Trolox (Trolox) was used as the positive control group.

[0073] As a result, as shown in Figure 6, it was found that the cell viability of fibroblasts and keratinocytes, which was reduced by UV irradiation, was restored by the peptide having the amino acid sequence of SEQ ID NO: 1. From the above results, it was found that the peptide according to one embodiment contributes to the inhibition of UV-induced death of skin cells.

[0074] Example 6. Confirmation of antioxidant effect on skin cells induced by ultraviolet light In this example, the effect of a peptide according to one embodiment on the antioxidant effect in skin cells induced by ultraviolet light is confirmed by evaluating the level of reactive oxygen species in skin cells that is increased by ultraviolet light irradiation.

[0075] Specifically, NIH3T3 cells or HaCaT cells were cultured in a 6-well plate at 5 × 10 5 After seeding at a density of cells / well, the cells were cultured for 24 hours. The cells were then washed once with serum-free DMEM media, and the culture medium was replaced with the serum-free medium, to which the peptide having the amino acid sequence of SEQ ID NO: 1 was dispensed at 50 μM or 100 μM. The culture medium to which the peptide was dispensed was cultured in a CO2 incubator at 37°C for 1 hour, transferred to an E-TUBE, mixed with 100 μl of PBS, and dispensed into a well plate. Then, NIH3T3 cells were exposed to 6 J / cm irradiated using an ultraviolet irradiator (Vilber Lourmat, France). 2 HaCaT cells were exposed to 15 mJ / cm of UV light. 2 The well plate was then irradiated with ultraviolet light of 1000 nm. The PBS in the well plate was then removed, and 900 μL of the culture medium containing the peptide was added, followed by culturing for 24 hours in a CO2 incubator at 37°C. The culture was then treated with 2',7'-dichlorofluorescein diacetate (DCFG-DA), wrapped in foil, and then cultured for 30 minutes in a CO2 incubator at 37°C. The culture was then washed twice with PBS, and 500 μL of 1× trypsin / EDTA was added to obtain cells, followed by centrifugation. The centrifuged cells were washed with PBS, and the FL1 fluorescence value was measured via flow cytometry (FACS (BD Biosciences, USA)). In this example, a non-treated group was used as the control group, and a group containing Trolox (Trolox) was used as the positive control group.

[0076] As a result, as shown in Figure 7, it was found that the peptide having the amino acid sequence of SEQ ID NO: 1 reduces the reactive oxygen levels in fibroblasts and keratinocytes that are increased by UV irradiation. From the above results, it was found that the peptide according to one embodiment contributes to the antioxidant effect on skin cells induced by UV rays.

[0077] Formulation example 1. Softening lotion A softening lotion containing a peptide according to one embodiment and having the following composition was prepared by a method known in the art.

[0078] [Table 4]

[0079] Dosage form example 2: Nutritious cream A nutritional cream containing a peptide according to one embodiment and having the following composition was prepared by a method known in the art.

[0080] [Table 5]

[0081] Formulation example 3: Nutritious lotion A nutritious lotion containing the peptide according to one embodiment and having the following composition was prepared by a method known in the art.

[0082] [Table 6]

[0083] Dosage form example 4. Essence An essence containing a peptide according to one embodiment and having the following composition was prepared by a method known in the art.

[0084] [Table 7]

[0085] The above description of the present invention is for illustrative purposes only, and a person having ordinary skill in the art to which the present invention pertains will understand that the present invention can be easily modified into other specific forms without changing the technical concept or essential features of the present invention. Therefore, the above-described embodiments are to be understood as illustrative in all respects and not restrictive.

Claims

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

1.

2. 2. The peptide according to claim 1, wherein the N-terminus of the peptide is bound to any one of 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 C-terminus of the peptide is an amino group (-NH ２ ), tertiary alkyl groups and azides (-NHNH ２ 2. The peptide of claim 1, wherein the peptide is linked to any one of the protecting groups selected from the group consisting of:

4. The peptide according to claim 1, which exhibits one or more of the following characteristics: (a) promoting the proliferation of fibroblasts or keratinocytes; (b) enhanced expression of type I collagen (Col1a1), fibronectin, or elastin, which are components of the extracellular matrix; (c) enhanced expression of skin barrier factors sirtuin 1 (SIRT-1) or aquaporin 3 (AQP3); and (d) Inhibition of fibroblast or keratinocyte death, and reduction of reactive oxygen levels.

5. A cosmetic composition for improving skin conditions, comprising the peptide according to any one of claims 1 to 4 as an active ingredient.

6. The cosmetic composition according to claim 5 , wherein the improvement of skin condition is improvement of wrinkles, improvement of skin elasticity, wound recovery, strengthening of the skin barrier, or inhibition of skin aging.

7. The cosmetic composition according to claim 6 , wherein the skin aging is caused by ultraviolet rays.

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