Peptides having skin condition-improving activity and uses thereof

Abstract

The present application relates to a peptide having skin condition-improving activity and use thereof, and provides a peptide consisting of the amino acid sequence of SEQ ID NO: 1 and a cosmetic composition for skin condition improvement including the peptide consisting of the amino acid sequence of SEQ ID NO: 1 as an effective ingredient.

Description

TECHNICAL FIELD

[0001] The present application relates to a peptide having skin condition improving activity and use thereof. BACKGROUND

[0002] Human skin is continuously changing, and the most representative of them is the decrease in skin function and visual aesthetic due to aging. Aging causes wrinkles in the skin, and as representative factors of wrinkle formation, exposure to ultraviolet rays and decrease in collagen biosynthesis can be cited. Skin aging is roughly divided into endogenous aging due to genetic factors and exogenous aging due to external environmental factors such as sunlight. Among them, for exogenous aging, it is known that aging can be prevented, treated or delayed by removing active oxygen, proliferating fibroblasts, and promoting collagen biosynthesis, etc.

[0003] Collagen, which is the main component of extracelluar matrix, is the main matrix protein produced in fibroblasts in the skin. Collagen forms most of the organic matter in the skin, tendon, bone and teeth, and the content in the bone and skin (dermis) is particularly high. Such collagen decreases with age and photoaging due to ultraviolet irradiation, which is known to be closely related to the formation of skin wrinkles. Also, collagen plays an important role in wound treatment, and promotes collagen synthesis in damaged epithelium, thereby enabling rapid and complete recovery of wounds. Also, it has been reported that as the biosynthesis of collagen is promoted and the density of the basement layer or the like is increased, the concentration of melanin per unit skin density decreases, and thus a skin whitening effect can be expected.

[0004] In this technical background, various studies for improving skin condition by promoting collagen biosynthesis, proliferating fibroblasts, etc., and increasing activity mechanisms are currently being conducted (Korean Registered Patent No. 10-1813629), but are still not perfect. SUMMARY

[0005] TECHNICAL PROBLEM

[0006] In one aspect, there is provided a peptide consisting of the amino acid sequence of SEQ ID NO: 1.

[0007] In another aspect, there is provided a cosmetic composition for skin condition improvement, including a peptide consisting of the amino acid sequence of SEQ ID NO: 1 as an effective ingredient.

[0008] Other objects and advantages of the present application will become more apparent in light of the following claims and the following detailed description of the application. As long as what is not described in the present specification is sufficiently recognized and inferred by those skilled in the art or those skilled in similar technical fields, the relevant description thereof is omitted.

[0009]

Technical Solution

[0010] Various explanations and embodiments disclosed in the present application can also be applied to various other explanations and embodiments. That is, all combinations of various elements disclosed in the present application belong to the scope of the present application. Also, it cannot be determined that the scope of the present application is limited to the specific recitations described below.

[0011] In one aspect, there is provided a peptide consisting of the amino acid sequence of SEQ ID NO: 1.

[0012] The term "peptide" used in the present specification can mean a linear molecule in which a plurality of amino acid residues are combined with each other by a peptide bond. The peptide can be prepared by a chemical synthesis method well known in the art, particularly, a solid phase synthesis technique or a liquid phase synthesis technique (US Registered Patent No. 5,516,891). As a result of efforts to develop a peptide having a biologically effective activity, the present inventors have identified a peptide consisting of the amino acid sequence of SEQ ID NO: 1. Here, the biologically effective activity can mean any one selected from the group consisting of (a) promoting proliferation of fibroblasts or keratinocytes; (b) increasing expression of collagen type I (Col1a1), fibronectin, or elastin as a constituent factor of extracellular matrix; (c) increasing expression of skin barrier factors, i.e., Sirtuin-1 (SIRT-1) or Aquaporin-3 (AQP3); and (d) inhibiting apoptosis of fibroblasts or keratinocytes and reducing active oxygen level. Accordingly, the peptide can be used for skin state improvement or antioxidant.

[0013] In the peptide, in order to obtain chemical stability, enhanced pharmacological properties (half-life, absorbability, potency, efficacy, etc.), altered specificity (e.g., a wide range of biological activity), reduced antigenicity, a protecting group can be bound at the N- or C-terminus of the terminal end of the peptide. In one embodiment, the N-terminus of the peptide can be bound with any one protecting group selected from the group consisting of an acetyl group, a fluorenylmethoxycarbonyl group, a formyl group, a palmitoyl group, a myristyl group, a stearyl group, a butoxycarbonyl group, an allyloxycarbonyl group, and a polyethylene glycol (PEG); and / or the C-terminus of the peptide can be bound with any one protecting group selected from the group consisting of an amino group (-NH2), a tertiary alkyl group, and a hydrazino group (-NHNH2). Also, the peptide can optionally further include a targeting sequence, a tag, a labeled residue, an amino acid sequence prepared for a specific purpose of increasing half-life or peptide stability.

[0014] The peptide is artificially synthesized, or non-naturally occurring or engineered, which means a state generated by performing artificial modification, rather than a state of existence as it is produced in a natural state. Here, the artificial modification can include artificial synthesis of an amino acid sequence by mimicking a plurality of amino acid structures, or can include engineering as above to obtain chemical stability, enhanced pharmacological properties, altered specificity, or reduced antigenicity.

[0015] The term "stability" used in the present specification can not only mean the in vivo stability of the peptide protected from the attack of in vivo proteolytic enzymes, but also can mean storage stability (e.g., room temperature storage stability).

[0016] Another aspect provides a cosmetic composition for skin condition improvement, including a peptide of an amino acid sequence of SEQ ID NO: 1 as an effective ingredient.

[0017] Among the terms or elements mentioned in the description of the peptide, the same as that already mentioned is as described above.

[0018] The term "improvement" used in the present specification can mean all actions related to alleviation or treatment of a state, such as alleviation of the degree of symptoms.

[0019] The term "skin state improvement" used in the present specification can broadly mean a process of treating, alleviating, or alleviating skin damage due to an endogenous factor of the skin or an exogenous factor of the skin, or the effect thereof, and can be interpreted as improvement of wrinkles, improvement of skin elasticity, recovery of a wound, strengthening of a skin barrier, or inhibition of skin aging, but is not limited thereto.

[0020] Here, "improvement of wrinkles", "improvement of skin elasticity", and "recovery of a wound" can mean all actions of increasing the total amount of extracellular matrix factors, including promotion of collagen synthesis, and the like. Also, "strengthening of a skin barrier" can mean strengthening of the inherent function of the skin, that is, prevention of leakage of moisture and nutrient components in the skin to the outside and prevention of invasion of harmful substances such as bacteria or viruses into the skin. Also, "inhibition of skin aging" can mean inhibition of wrinkles, skin sagging, reduction of elasticity, and the like, which are reduction of skin functions. At this time, the skin aging can be photoaging, for example, skin aging due to ultraviolet rays.

[0021] The existing functional peptides have a disadvantage in that, even though they have effective biological activities, they cannot be effectively introduced into target tissues or cells due to the size of the peptides themselves, or are rapidly disappeared from the body in a short time due to a short half-life. In contrast, the cosmetic composition according to an embodiment includes a peptide consisting of 10 or less amino acids as an effective ingredient, and thus, the skin permeability or the like of the effective ingredient is very excellent, and for example, when applied topically to the skin, it can effectively improve the skin state.

[0022] According to an embodiment, it is possible to promote the proliferation of fibroblasts and keratinocytes and to enhance the synthesis of extracellular matrix constituting factors and skin barrier factors. Also, it was found that the peptide can restore the reduced activity of fibroblasts and keratinocytes and can enhance their antioxidant efficacy. Thus, the peptide can be used as an effective ingredient of a cosmetic composition for skin state improvement.

[0023] The cosmetic composition can include a cosmetically effective amount of the peptide; and / or a cosmetically acceptable carrier, but is not limited thereto.

[0024] The term "cosmetically effective amount" used in the present specification means an amount sufficient to achieve the skin state improvement efficacy of the cosmetic composition.

[0025] The weight ratio between the peptide and the cosmetically acceptable carrier can be, for example, 500:1 to 1:500, for example, the weight ratio can be 450:1 to 1:450, 400:1 to 1:400, 350:1 to 1:350, 300:1 to 1:300, 250:1 to 1:250, 200:1 to 1:200, 150:1 to 1:150, 100:1 to 1:100, 80:1 to 1:80, 60:1 to 1:60, 40:1 to 1:40, 20:1 to 1:20, 10:1 to 1:10, 8:1 to 1:8, 6:1 to 1:6, 4:1 to 1:4, or 2:1 to 1:2, but is not limited thereto.

[0026] The cosmetic composition can be prepared in any dosage form generally prepared in the art, for example, can be prepared as a solution, a suspension, an emulsion, a dough, a gel, a skin cream, a skin lotion, a powder, a soap, a surfactant-containing cleanser, an oil, a powder foundation, a foundation lotion, a wax foundation, and a spray, etc., but is not limited thereto. For example, the cosmetic composition can be prepared as a softening toner, a nourishing toner, a nourishing cream, a massage cream, an essence, an eye cream, a cleansing cream, a cleansing bubble, a cleansing water, a pack, a spray, or a powder dosage form.

[0027] In the case where the dosage form of the cosmetic composition is a dough, a skin cream, or a gel, an animal oil, a vegetable oil, a wax, a paraffin, a starch, a tragacanth gum, a cellulose derivative, a polyethylene glycol, a silicon, a bentonite, a silicon dioxide, a talc, or a zinc oxide, etc. can be used as a carrier ingredient.

[0028] In the case where the dosage form of the cosmetic composition is a foundation or a spray, lactose, talc, silicon dioxide, aluminum hydroxide, calcium silicate, or a polyamide powder can be used as a carrier ingredient, for example, if it is a spray, a propellant such as chlorofluorocarbon, propane / butane, or dimethyl ether can be further included.

[0029] In the case where the dosage form of the cosmetic composition is a solution or an emulsion, a solvent, a solubilizer, or an emulsifier can be used as a carrier ingredient, for example, water, ethanol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol, or sorbitan fatty acid ester can be included.

[0030] In the case where the dosage form of the cosmetic composition is a suspension, a liquid phase diluent such as water, ethanol, or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, and polyoxyethylene sorbitol ester, microcrystalline cellulose, methyl aluminum hydroxide, bentonite, agar, or tragacanth gum, etc. can be used.

[0031] In the case where the dosage form of the cosmetic composition is a surfactant-containing cleanser, a fatty alcohol sulfate, a fatty alcohol ether sulfate, a sulfosuccinic monoester, a isethionate, an imidazoline derivative, a methyltaurinate, a sarcosinate, a fatty acid amide ether sulfate, an alkyl amido betaine, a fatty alcohol, a fatty acid glyceride, a fatty acid diethanolamide, a vegetable oil, a lanolin derivative, or an ethoxylated glycerol fatty acid ester, etc. can be used as the carrier ingredient.

[0032] The peptide can be included in a nanosome or nanoparticle to further improve the skin penetration problem or the stability problem. For example, the nanosome can be prepared using lecithin as a raw material with a microfluidizer, and can be included in a lecithin particle. As a method of preparing the nanosome, any known method can be used. The size of the nanosome particle is preferably 30 to 200 nm. In the case where the size of the nanosome particle is less than 30 nm, the skin penetration can be very fast to cause a skin side effect to occur, and in the case where the size of the nanosome particle is greater than 200 nm, it can be difficult to penetrate into the skin to make it difficult to obtain the use effect of the nanosome.

[0033] In addition to the peptide as the active ingredient and the carrier ingredient, the ingredients included in the cosmetic composition include the conventional ingredients for the cosmetic composition, for example, the conventional adjuvants such as an antioxidant, a stabilizer, a solubilizer, a vitamin, a pigment, and a perfume can be included.

[0034] The content of the peptide as the active ingredient included in the cosmetic composition can be appropriately and non-limitingly selected depending on the product form, the desired use, etc., for example, can be added in an amount of 0.01 to 15% by weight based on the total weight of the cosmetic composition. Also, for example, the cosmetic composition can include 1.0 to 3.0% by weight, preferably 2.0 to 3.0% by weight of the peptide based on the total weight.

[0035] On the other hand, there is provided a method of improving the skin state, including the step of applying a cosmetic composition including a peptide consisting of the amino acid sequence of SEQ ID NO: 1 as an active ingredient to the skin of an individual; a use of the peptide consisting of the amino acid sequence of SEQ ID NO: 1 in the preparation of a composition for improving the skin state.

[0036] Among the terms or elements mentioned in the description regarding the cosmetic composition, the same as that which has been mentioned is as described above.

[0037] The term "individual" used in the present specification means a subject in need of improvement of the skin state, and more specifically, a human or a non-human primate, a mouse, a dog, a cat, a horse, a cow, and the like mammal.

[0038] The terms "application", "administration" and "coating" used in the present specification can be used interchangeably, and can mean to localize at least partially a composition according to an embodiment on a desired site, or to dispose a composition according to an embodiment in a subject through an administration route.

[0039] Another aspect provides an antioxidant composition including a peptide consisting of the amino acid sequence of SEQ ID NO: 1 as an effective ingredient.

[0040] Among the terms or elements mentioned in the description of the peptide, the same as that already mentioned is as described above.

[0041] The antioxidant composition can be in the form of a pharmaceutical composition, a non-pharmaceutical composition, or a cosmetic composition, and as an example, the composition can be used as a cosmetic composition for skin condition improvement, or a pharmaceutical composition for improvement or treatment of a skin damage-related disease condition.

[0042] According to an embodiment, it is indicated that the peptide is capable of restoring the reduced activity of fibroblasts or keratinocytes and enhancing the antioxidant effect thereof. Thus, the peptide can be used as an effective ingredient of an antioxidant composition.

[0043] As an example, the antioxidant composition can be provided in the form of a pharmaceutical composition. The pharmaceutical composition can include a pharmaceutically effective amount of the peptide; and / or a pharmaceutically acceptable carrier, but is not limited thereto.

[0044] The term "pharmaceutically effective amount" used in the present specification can mean an amount of the pharmaceutical composition sufficient to achieve a prophylactic or therapeutic effect on a skin damage-related disease.

[0045] The pharmaceutically acceptable carrier is generally used in preparation, and includes lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinyl pyrrolidone, cellulose, water, sugar syrup, methyl cellulose, glycerol, hydroxybenzoate, hydroxypropyl benzoate, talc, magnesium stearate, and mineral oil, etc., but is not limited thereto. Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington's Pharmaceutical Sciences (19th edition, 1995).

[0046] The weight ratio between the peptide and the pharmaceutically acceptable carrier can be, for example, 500:1 to 1:500, and as an example, the weight ratio can be 450:1 to 1:450, 400:1 to 1:400, 350:1 to 1:350, 300:1 to 1:300, 250:1 to 1:250, 200:1 to 1:200, 150:1 to 1:150, 100:1 to 1:100, 80:1 to 1:80, 60:1 to 1:60, 40:1 to 1:40, 20:1 to 1:20, 10:1 to 1:10, 8:1 to 1:8, 6:1 to 1:6, 4:1 to 1:4, or 2:1 to 1:2, but is not limited thereto.

[0047] In addition to the ingredients, the pharmaceutical composition can further include a lubricant, a humectant, a sweetener, a flavoring agent, an emulsifier, a suspending agent, a preservative, etc., but is not limited thereto.

[0048] The pharmaceutical composition can be administered by enteral or parenteral, preferably by parenteral, and for parenteral administration, can be administered by intramuscular injection, intravenous injection, subcutaneous injection, intraperitoneal injection, topical administration, transdermal administration, etc., but is not limited thereto.

[0049] The administration amount of the pharmaceutical composition can be 0.0001 to 1000 μg (microgram), 0.001 to 1000 μg, 0.01 to 1000 μg, 0.1 to 1000 μg, or 1.0 to 1000 μg per day, but is not limited thereto, and can be prescribed differently according to the formulation method, the administration method, the age, the weight, the gender, the condition, the diet, the administration time, the administration route, the excretion rate, and the reaction sensitivity of the patient, etc.

[0050] The pharmaceutical composition can be prepared in a unit capacity form using a pharmaceutically acceptable carrier and / or an excipient according to a method easily performed by one of ordinary skill in the art to which the present application pertains, or can be prepared by being injected into a multi-capacity container.

[0051] The dosage form can be in the form of a solution, a suspension, or an emulsion in an oily or aqueous solvent, can also have the form of an extractant, a powder, a granule, a tablet, or a capsule, and can further include a powder and / or a stabilizer.

[0052] Another aspect provides a food composition for improving the skin condition, including a peptide consisting of the amino acid sequence of SEQ ID NO: 1 as an effective ingredient.

[0053] Among the terms or elements mentioned in the description regarding the peptide, the same as that which has been mentioned is as described above.

[0054] The content of peptides as active ingredients included in the food composition may be appropriately and non-limitingly selected according to the food form, intended use, etc., for example, they may be added at a rate of 0.01 to 15% by weight of the total food. For example, for a health beverage composition, they may be added at a rate of 0.02 to 10g, preferably 0.3 to 1g, based on 100ml.

[0055] [Beneficial Effects]

[0056] By targeting a specific peptide, it is possible to promote the proliferation of fibroblasts and keratinocytes and enhance the synthesis of extracellular matrix constituents and skin barrier factors. Therefore, it can be applied to improve skin condition, including wrinkle improvement, skin elasticity improvement, wound healing, skin barrier strengthening, or skin aging inhibition.

[0057] By targeting a specific peptide, it is possible to restore the reduced activity of fibroblasts and keratinocytes and enhance their antioxidant effects, thereby helping to restore skin damage that may be caused by external environmental factors such as ultraviolet radiation.

[0058] Therefore, peptides can be used as active ingredients in compositions for improving skin condition or in antioxidant compositions. [Attached Image Description]

[0059] Figure 1 The results show the cell proliferation level confirmed by changes in cell viability after a peptide consisting of the amino acid sequence of SEQ ID NO: 1 was added to NIH3T3 cells.

[0060] Figure 2 The results show that adding a peptide consisting of the amino acid sequence of SEQ ID NO: 1 to NIH3T3 cells confirmed an increase in the expression of extracellular matrix components, namely type I collagen (Col1a1), fibronectin, and elastin.

[0061] Figure 3 This demonstrates the quantitative assessment of the expression levels of extracellular matrix components after adding a peptide consisting of the amino acid sequence of SEQ ID NO: 1 to NIH3T3 cells. Figure 3 A shows the results confirming the expression level of Col1a1. Figure 3 B shows the results confirming fibronectin expression levels. Figure 3 C indicates the results confirming the level of elastin expression.

[0062] Figure 4 The results show that adding the peptide consisting of the amino acid sequence of SEQ ID NO: 1 to HaCaT cells confirmed an increase in the expression of skin barrier factors, namely silencing regulator protein 1 (SIRT-1) and aquaporin 3 (AQP3).

[0063] Figure 5 This demonstrates the quantitative assessment of skin barrier factor expression levels after the peptide consisting of the amino acid sequence of SEQ ID NO: 1 was added to HaCaT cells. Figure 5 A shows the results confirming SIRT-1 expression levels. Figure 5 B shows the results confirming the AQP3 expression level.

[0064] Figure 6 The results show that adding a peptide consisting of the amino acid sequence of SEQ ID NO: 1 to NIH3T3 cells confirmed the inhibitory effect on skin cell apoptosis induced by ultraviolet irradiation by survival assessment.

[0065] Figure 7 The results show that the addition of a peptide consisting of the amino acid sequence of SEQ ID NO: 1 to HaCaT cells confirmed the inhibitory effect on skin cell apoptosis induced by ultraviolet irradiation by survival assessment.

[0066] Figure 8 The results show the quantitative confirmation of changes in reactive oxygen species levels due to ultraviolet radiation after the addition of a peptide consisting of the amino acid sequence of SEQ ID NO: 1 to NIH3T3 cells.

[0067] Figure 9 This demonstrates the results of quantitative confirmation of changes in reactive oxygen species levels due to ultraviolet radiation after the addition of a peptide consisting of the amino acid sequence of SEQ ID NO: 1 to HaCaT cells.

Detailed Implementation Methods

[0068] The present invention will now be described in more detail through embodiments. However, these embodiments are merely illustrative of the invention, and the scope of the invention is not limited to these embodiments.

[0069] Example 1: Peptide Synthesis

[0070] Peptides with the amino acid sequence SEQ ID NO: 1, as described in Table 1 below, were synthesized using an automated peptide synthesizer (Milligen 9050, Millipore, USA). The synthesized peptides were purified using a C18 reversed-phase high-performance liquid chromatography (HPLC) system (Waters Associates, USA). An ACQUITY UPLC BEH300 C18 column (2.1 mm × 100 mm, 1.7 μm, Waters Co., USA) was used.

[0071] Table 1

[0072] SEQ ID NO: Sequence (N-terminal → C-terminal) 1 EVVDI

[0073] Example 2: Confirmation of the proliferation promotion effect on skin cells

[0074] This example aims to confirm the effect of the peptide according to an embodiment on the improvement of endogenous skin aging including wrinkle improvement and elasticity enhancement by evaluating the change in survival rate of mouse fibroblast cells, i.e., NIH3T3 cells.

[0075] Specifically, NIH3T3 cells were seeded at a density of 1 x 10 4 cells / well in a 96-well plate and cultured for 24 hours. Then, the cells were washed once with serum-free DMEM media, and a peptide consisting of the amino acid sequence of SEQ ID NO: 1 was aliquoted at 50 μM or 100 μM in 200 μL of the serum-free DMEM media and cultured in a CO2 incubator at 37°C for 72 hours. Then, after washing the culture twice with PBS, an MTT solution was aliquoted at 0.5 mg / ml in each well. Then, after shielding from light, it was cultured in a CO2 incubator at 37°C for 4 hours, and the absorbance at 540 nm was measured using a microplate reader (Molecular Devices, USA).

[0076] As a result, it was confirmed that the proliferation of fibroblasts was promoted by the peptide consisting of the amino acid sequence of SEQ ID NO: 1 as shown in Table 1. Figure 1

[0077] Example 3: Confirmation of wrinkle improvement and elasticity enhancement effect

[0078] This example aims to confirm the effect of the peptide according to an embodiment on the improvement of endogenous skin aging including wrinkle improvement and elasticity enhancement by evaluating the expression level of collagen type I (Col1a1), fibronectin, or elastin known as a component of the dermis.

[0079] Specifically, NIH3T3 cells were seeded at a density of 3 x 10 5 ​The cells were seeded at a density of 1 x 105 cells / well in a 6-well plate and cultured for 24 hours. Then, after washing the cells once with serum-free DMEM media, a peptide consisting of the amino acid sequence of SEQ ID NO: 1 was added to 1 mL of the culture medium at 10 μM or 50 μM, and was cultured in a CO2 incubator at 37°C for 24 hours. After 24 hours, after washing the culture twice with PBS, easy blue (iNtRON, Cat. No: 17061, Korea) was used to isolate ribonucleic acid (RNA) from the culture product. Then, reverse transcription was performed on the isolated RNA using an RT kit (Enzynomics, Cat. NO: RT200, Korea) to synthesize complementary deoxyribonucleic acid (cDNA). Then, a polymerase chain reaction (PCR) was performed using 1 μg of the synthesized cDNA and primers for Col1a1, fibronectin, or elastin and a PCR kit (enzynomics, Cat. No: P581T, Korea). In this example, the control group was an untreated group, and a group to which transforming growth factor-β1 (TGF-β1) was added was used as a positive control group, and the nucleotide sequences of the primers used in this example are as follows in Table 2.

[0080] [Table 2]

[0081]

[0082]

[0083] As a result, it was confirmed that, as shown in FIGS. 1 to 4, due to the peptide consisting of the amino acid sequence of SEQ ID NO: 1, the expression of extracellular matrix components, i.e., Col1a1, fibronectin, and elastin, was increased. Figure 2 3

[0084] [Example 4: Confirmation of skin barrier strengthening effect]

[0085] This example aims to confirm the effect of the peptide according to an embodiment on skin barrier strengthening by evaluating the expression level of sirtuin 1 (SIRT-1) or aquaporin 3 (AQP3).

[0086] Specifically, HaCaT cells were seeded at a density of 3 x 105 cells / well in a 6-well plate and cultured for 24 hours. Then, after washing the cells once with serum-free DMEM media, a peptide consisting of the amino acid sequence of SEQ ID NO: 1 was added to 1 mL of the culture medium at 10 μM or 50 μM, and was cultured in a CO2 incubator at 37°C for 24 hours. After 24 hours, after washing the culture twice with PBS, easy blue (iNtRON, Cat. No: 17061, Korea) was used to isolate ribonucleic acid (RNA) from the culture product. Then, reverse transcription was performed on the isolated RNA using an RT kit (Enzynomics, Cat. NO: RT200, Korea) to synthesize complementary deoxyribonucleic acid (cDNA). Then, a polymerase chain reaction (PCR) was performed using 1 μg of the synthesized cDNA and primers for Col1a1, fibronectin, or elastin and a PCR kit (enzynomics, Cat. No: P581T, Korea). In this example, the control group was an untreated group, and a group to which transforming growth factor-β1 (TGF-β1) was added was used as a positive control group, and the nucleotide sequences of the primers used in this example are as follows in Table 2. 5 ​​The cells were seeded at a density of 1 x 105cells / well in a 6-well plate and cultured for 24 hours. Then, after washing the cells once with serum-free DMEM media, a peptide consisting of the amino acid sequence of SEQ ID NO: 1 was subcultured at 50 μM or 100 μM in 1 mL of the culture medium and cultured in a CO2incubator at 37°C for 24 hours. Then, after washing the culture twice with PBS, easy blue (iNtRON, Cat. No: 17061, Korea) was used to isolate ribonucleic acid (RNA) from the culture product. Then, reverse transcription was performed on the isolated RNA using an RT kit (Enzynomics, Cat. NO: RT200, Korea) to synthesize complementary deoxyribonucleic acid (cDNA). Then, a polymerase chain reaction (PCR) was performed using 1 μg of the synthesized cDNA and primers for SIRT-1 or AQP3 and a PCR kit (enzynomics, Cat. No.: P581T, Korea). In this example, the control group was an untreated group, and a group to which EGF was added was used as a positive control group. The nucleotide sequences of the primers used in this example are shown in Table 3 below.

[0087] [Table 3]

[0088]

[0089] As a result, it was confirmed that, as shown in Figure 4 and Figure 5 , the expression of SIRT-1 and AQP3, which are skin barrier factors, was increased due to the peptide consisting of the amino acid sequence of SEQ ID NO: 1. From the results, it can be seen that the peptide according to an embodiment contributes to skin barrier strengthening and skin anti-aging by increasing skin barrier factors.

[0090] [Example 5: Confirmation of the inhibitory effect on skin cell apoptosis induced by ultraviolet rays]

[0091] This example aims to confirm the effect of the peptide according to an embodiment on the inhibition or recovery of skin cell apoptosis induced by ultraviolet rays by evaluating the change in the survival rate of skin cells in which apoptosis was induced by irradiation of ultraviolet rays.

[0092] Specifically, NIH3T3 cells or HaCaT cells were seeded at a density of 1 x 105cells / well in a 6-well plate and cultured for 24 hours. Then, after washing the cells once with serum-free DMEM media, a peptide consisting of the amino acid sequence of SEQ ID NO: 1 was subcultured at 50 μM or 100 μM in 1 mL of the culture medium and cultured in a CO2incubator at 37°C for 24 hours. Then, after washing the culture twice with PBS, easy blue (iNtRON, Cat. No: 17061, Korea) was used to isolate ribonucleic acid (RNA) from the culture product. Then, reverse transcription was performed on the isolated RNA using an RT kit (Enzynomics, Cat. NO: RT200, Korea) to synthesize complementary deoxyribonucleic acid (cDNA). Then, a polymerase chain reaction (PCR) was performed using 1 μg of the synthesized cDNA and primers for SIRT-1 or AQP3 and a PCR kit (enzynomics, Cat. No.: P581T, Korea). In this example, the control group was an untreated group, and a group to which EGF was added was used as a positive control group. The nucleotide sequences of the primers used in this example are shown in Table 3 below. 4Cells were seeded at a density of 1:1 in 96-well plates and cultured for 24 hours. The cells were then washed once with serum-free DMEM media, and the peptide consisting of the amino acid sequence of SEQ ID NO: 1 was divided into 50 μM or 100 μM solutions.

[0093] Prepare 200 μL of culture medium. Incubate the culture medium containing the peptide isolates in a CO2 incubator at 37°C for 1 hour. Transfer the medium to an e-tube and mix with 100 μL of PBS to isolate the isolates into wells of a plate. Then, irradiate NIH3T3 cells with a UV irradiator (VILBER LOURMAT, France) at 6 J / cm². 2 Ultraviolet light was used to irradiate HaCaT cells at 15 mJ / cm². 2 The culture was then exposed to ultraviolet light. The PBS was removed from the wells, and MTT (Sigma, Cat. No.: M2003, USA) solution was aliquoted at 0.5 mg / ml into each well. The culture was then incubated at 37°C in a CO2 incubator for 72 hours. The absorbance of the culture 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 group treated with Trolox (Trolox) served as the positive control group.

[0094] As a result, it was confirmed that... Figure 6 and 7 As shown, a peptide consisting of the amino acid sequence of SEQ ID NO: 1 restored the cell viability of fibroblasts and keratinocytes reduced by ultraviolet radiation. The results indicate that the peptide according to one embodiment helps to inhibit ultraviolet-induced apoptosis of skin cells.

[0095] [Example 6: Confirmation of the reduction effect of increased reactive oxygen species due to ultraviolet radiation]

[0096] This embodiment aims to confirm the effect of a peptide according to one embodiment on the antioxidant effect in damaged skin cells by evaluating changes in the level of reactive oxygen species in skin cells due to increased exposure to ultraviolet radiation.

[0097] Specifically, NIH3T3 cells or HaCaT cells were administered at a rate of 5 × 10⁻⁶. 5The cells were seeded at a density of 1 cell per well in a 6-well plate and then cultured for 24 hours. Then, after washing the cells once with serum-free DMEM media, a peptide consisting of the amino acid sequence of SEQ ID NO: 1 was added to 1 mL of the culture medium at a concentration of 50 μM or 100 μM, and the mixture was cultured in a CO2 incubator at 37°C for 1 hour. Then, the culture was moved to an e-tube and mixed with 1 mL of PBS to be dispensed into the well plate. Then, the NIH3T3 cells were irradiated with UV light at 6 J / cm2and the HaCaT cells were irradiated with UV light at 15 mJ / cm2using a UV irradiator (VILBER LOURMAT, France). Then, the PBS in the well plate was removed, and 900 μL of the culture medium to which the peptide was added was added, and the mixture was cultured in a CO2 incubator at 37°C for 24 hours. Then, the culture product was treated with 10 μM of 2',7'-dichlorofluorescin diacetate (DCFG-DA), the culture product was wrapped with foil, and the culture product was cultured in a CO2 incubator at 37°C for 30 minutes. Then, the culture product was washed twice with PBS and treated with 500 μL of 1x trypsin / ethylenediaminetetraacetic acid (EDTA) to obtain cells, and then centrifuged. The centrifuged cells were washed with PBS and the FL1 fluorescence value was measured using flow cytometry (FACS, BD, USA). In this example, the control group was the untreated group, the comparison group was the group irradiated with UV light only, and the group to which Trolox was added (Trolox) was used as a positive control group. 2 2 As a result, it was confirmed that the level of reactive oxygen species in fibroblasts and keratinocytes increased by irradiation with UV light was reduced by the peptide consisting of the amino acid sequence of SEQ ID NO: 1, as shown in

[0098] Figure 8 9 According to the results, it was confirmed that the level of reactive oxygen species in fibroblasts and keratinocytes increased by irradiation with UV light was reduced by the peptide consisting of the amino acid sequence of SEQ ID NO: 1, as shown in

[0099]

Dosage Form Example 1: Preparation of Peptide Nanosomes

[0100] The peptide of Example 1, 50 mg, was dissolved by stirring in 500 mL of distilled water. The complex solution was mixed with lecithin 5 g, sodium oleate 0.3 mL, ethanol 50 mL, and a small amount of an oil phase, and then distilled water was added to adjust the amount to a total of 1 L, and then emulsification was performed using a high-pressure microfluidizer at high pressure to prepare peptide nanosomes having a size of about 100 nm.​​​

[0101] Example 2: Softening lotion

[0102] A softening lotion including a peptide according to an embodiment and having the following composition was prepared in a manner known in the art.

[0103] Table 4

[0104]

[0105]

[0106] Example 3: Nutrient cream

[0107] A nutrient cream including a peptide according to an embodiment and having the following composition was prepared in a manner known in the art.

[0108] Table 5

[0109] Ingredient Content (wt%) Peptide 2.5 Rapeseed oil 3 Cetostearyl alcohol 1.5 Stearic acid 1.5 Glycerol stearate 1.5 Liquid paraffin 10 Beeswax 2 Polysorbate 60 0.6 Sorbitan sesquioleate 2.5 Squalene 3 1,3-Butylene glycol 3 Glycerol 5 Triethanolamine 0.5 Tocopheryl acetate 0.5 Preservatives, pigments q.s. Fragrance q.s. Purified water q.s. Total 100

[0110] Example 4: Nutrient lotion

[0111] A nutrient lotion including a peptide according to an embodiment and having the following composition was prepared in a manner known in the art.

[0112] Table 6

[0113] Ingredient Content (wt%) Peptide 3.0 1,3-Butylene glycol 4 Glycerol 4 Cetostearyl alcohol 0.8 Glycerol stearate 1 Triethanolamine 0.13 Tocopheryl acetate 0.3 Liquid paraffin 5 Squalene 3 Oil 2 Polysorbate 60 1.5 Sorbitan sesquioleate 0.5 Carboxyvinyl polymer 1 Preservatives, pigments q.s. Fragrance q.s. Purified water q.s. Total 100

[0114] Example 5: Serum

[0115] A serum including a peptide according to an embodiment and having the following composition was prepared in a manner known in the art.

[0116] Table 7

[0117]

[0118]

[0119] The above description is merely illustrative of the application and not restrictive. Since the application can be modified in various ways and we therefore intend to be bound only by the application covered by the scope of the patent claims, it should be understood that the application can be practiced otherwise than as specifically described.

Claims

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

1.

2. The peptide according to claim 1, wherein, The N-terminus of the peptide is bound to any one of the protecting groups selected from the group consisting of acetyl, palmitoyl, myristoyl, stearoyl, and polyethylene glycol.

3. The peptide according to claim 1, wherein, The C-terminus of the peptide is bound to an amino group.

4. A cosmetic composition for improving skin condition, comprising a peptide as an active ingredient according to any one of claims 1 to 3.

5. The cosmetic composition according to claim 4, wherein, The peptide was prepared in the form of nanobody.

6. Use of the peptide according to any one of claims 1 to 3 in the preparation of a cosmetic composition for improving skin condition.

7. The use according to claim 6, wherein, The improvement in skin condition is described as wound healing, enhanced skin barrier, or inhibition of skin aging.

8. The use according to claim 6, wherein, The improvement in skin condition refers to the reduction of wrinkles.

9. The use according to claim 6, wherein, The improvement in skin condition refers to improved skin elasticity.

10. The use according to claim 7, wherein, The skin aging mentioned refers to skin aging caused by ultraviolet radiation.

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