WO2019117577A1 - Anti-aging or anti-inflammatory composition comprising kisspeptin - Google Patents

Abstract

The present invention relates to a composition comprising kisspeptin. Kisspeptin or an active fragment thereof according to an aspect of the present invention has activity to inhibit MMP-1 or inflammatory cytokines, and thus can be favorably used in the prevention, alleviation, or treatment of skin aging, skin damage, skin conditions, or inflammatory diseases.

Description

An anti-aging or anti-inflammatory composition comprising kispeptin

Kispeptin. ≪ / RTI >

Functional cosmetics may include products that delay skin aging and help to improve wrinkles of the skin, products that protect the skin from ultraviolet rays and the external environment, or products that help to whiten skin. For the development of functional cosmetics, it is necessary to develop functional ingredients that can exhibit physiological efficacy in the skin.

The human skin undergoes changes due to various internal and external factors as it gets older. In other words, the secretion of various hormones that regulate metabolism is reduced internally, and the function of immune cells and the activity of cells are lowered, so that the biosynthesis of immune proteins and biocompatible proteins necessary for the living body is reduced. As a result of the destruction of the ozone layer, the amount of ultraviolet rays reaching the surface of the sunlight increases and the environmental pollution is further increased. As a result, free radicals and active noxious oxygen are increased and the thickness of the skin decreases, Not only the elasticity is reduced but also the skin color is gray, the skin trouble frequently occurs, and the spots, the freckles and the black spots are also increased.

Detailed mechanism of skin aging phenomenon is as follows: reduction of keratinocyte and fibroblast cleavage, reduction of collagen synthesis, increase of MMP (Matrix metalloproteinase) generation, increase of signaling of melanin generation, The elasticity of the skin decreases, and the number of spots, freckles, and black spots increases. In particular, the balance of hormones in the forties is broken, and cell regeneration ability, collagen synthesis ability, and skin lipid production ability, which plays an important role in skin moisturization, are greatly reduced. In addition, during the first five years after menopause, collagen is reduced by about 30%, the abnormal tangling phenomenon of these collagen fibers is increased, skin elasticity is significantly lowered, and hyaluronic acid, which is responsible for moisture in the skin, It is known that the amount of production of the can is significantly reduced.

On the other hand, ultraviolet light activates stress hormones repeatedly and damages the skin. This can lead to skin side effects such as the collapse of dermal fibrous tissue, just as overdose of steroids on skin. UV stimulation is a major cause of glucocorticoid activity in human skin known as stress hormones such as cortisol. It also increases the expression of 11? -Hydroxysteroid dehydrogenase type 1 (11? -HSD1), an enzyme in the body that activates cortisol. The increase in the activity and expression of 11? -HSD1 by ultraviolet rays is closely related to skin aging, and thus has been attracting attention as a new target for inhibiting skin aging.

Inflammation is a physiological response that protects the body from harmful environmental conditions, such as the ingress of external foreign matter, such as bacteria, and mechanical damage. Excessive inflammation is also a major cause of skin damage. Inflammation causes an increase in several types of polynuclear leukocytes (PMNs) and immunological substances, and these cells contain various types of proteolytic enzymes, such as inflammatory cell products (such as elastase, hyalurinidase, and lipoxygenase ) And cytokine secretion. These actions may then cause harmful damage to adjacent tissue and non-tissue cell components and may lead to severe tissue damage of chronic inflammation. In addition, the damage of the connective tissues not only causes a decrease in the elasticity of the skin to cause wrinkles but also adversely affects the regeneration and proliferation of the cells, resulting in quick skin aging.

Various methods for improving the skin aging phenomenon and inflammation have been applied. Recently, various proteins have been studied for recovering skin damage as described above. Recently, kisspeptin protein has been actively studied as a regulatory agent for sexual maturation and cancer inhibitor. However, researches and experiments on skin aging and inflammation due to kisspeptin expression are insignificant.

One aspect is to provide a cosmetic composition comprising kisspeptin or an active fragment thereof as an active ingredient.

Another aspect is to provide a use for the use of kispeptin or an active fragment thereof in the preparation of a cosmetic composition.

Another aspect is to provide a composition for external application for skin for preventing, ameliorating or treating a skin condition comprising kisspeptin or an active fragment thereof as an active ingredient, or preventing or ameliorating skin aging.

Another aspect is to provide a pharmaceutical composition for preventing or ameliorating skin aging comprising kispeptin or an active fragment thereof as an active ingredient, and for the prophylaxis and treatment of inflammatory diseases.

Another aspect is to provide a method of preventing, ameliorating or treating skin conditions comprising administering kisspeptin or an active fragment thereof to an individual in need thereof.

Another aspect is to provide a use of kisspeptin or an active fragment thereof for use in the inhibition or amelioration of skin aging, in the preparation of a pharmaceutical composition for the prophylaxis and treatment of inflammatory diseases.

Yet another aspect is to provide a health functional food for the prevention or treatment of skin conditions comprising kispeptin or an active fragment thereof, or for preventing or ameliorating skin aging.

Another aspect is the use of kisspeptin or active fragments thereof for use in the manufacture of a composition for preventing, ameliorating or treating skin conditions.

One aspect provides a composition comprising kisspeptin or an active fragment thereof as an active ingredient.

The term " Kisspeptin "is used interchangeably with metastin or KISS-1 peptides and can refer to a protein encoded by the KISS1 gene. Kisspeptin is a G-protein coupled receptor ligand for GPR54 and is known as a human metastasis inhibitory gene with the ability to inhibit melanoma and breast cancer metastasis. GpR54 signaling plays an important role in initiating the secretion of gonadotropin releasing hormone (GnRH) in puberty. Gonadotropin releasing hormone is released from the hypothalamus and acts on the anterior pituitary gland to produce luteinizing hormone (LH) and follicle It causes the release of stimulating hormone (FSH). Because kispeptin can directly stimulate GnRH release, it transmits negative and positive feedback signals of steroid hormones to GnRH neurons, acts as a gatekeeper to the onset of puberty, ) And photoperiodic information. Wherein the kisspeptin is at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% Or may have sequence identity.

The active fragment may be composed of the 27th to 54th, 45th to 49th, or 45th to 54th amino acid sequences from the N-terminal of the amino acid sequence of SEQ ID NO: 1. The C-terminus of the active fragment may be amidated. In one embodiment, the active fragment has at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96% , 97% or more, 98% or more, 99% or more, or 100% sequence identity.

Refers to or includes the reduction, progression or prevention of a disease, disorder, condition or condition, or one or more symptoms thereof, wherein the "active ingredient" or " May refer to any amount of composition used in the practice of the invention provided herein sufficient to alleviate, inhibit or prevent the condition or condition, or one or more symptoms thereof. The effective amount (or dose) may be from 0.0001 mg to 10,000 mg, 0.001 mg to 1000 mg, 1.0 mg to 100 mg, 0.01 mg to 1000 mg, 0.01 mg to 100 mg, 0.01 mg to 10 mg, or 0.01 mg to 1 mg. The kisspeptin or active fragment thereof may be present in an amount of from 0.001% to 80%, such as 0.01% to 60%, 0.01% to 40%, 0.01% to 30%, 0.01% From 0.05 wt% to 40 wt%, from 0.05 wt% to 30 wt%, from 0.05 wt% to 20 wt%, from 0.01 wt% to 10 wt%, from 0.01 wt% to 5 wt% From 0.05% to 10%, from 0.05% to 5%, from 0.1% to 60%, from 0.1% to 40%, from 0.1% to 30%, from 0.1% to 20% By weight, 0.1% by weight to 10% by weight, or 0.1% by weight to 5% by weight.

In one embodiment, the kispeptin or active fragment thereof may be one that has activity to inhibit MMP (Matrix metalloproteinase), ll [beta] -HSDl or an inflammatory factor such as inflammatory cytokine, IL-6, or IL-8 have. Therefore, the kispeptin or an active fragment thereof is effective for prevention, improvement, or treatment of skin aging, skin damage, skin condition or inflammatory disease.

Thus, the composition may be one for the prevention, amelioration or treatment of aging, for example, skin aging, skin damage, skin conditions or inflammatory diseases.

As used herein, the term "skin aging" refers to the type and intangible changes that appear on the skin with age, such as skin thinning, the number of dermal cells or blood vessels, DNA damage recovery, Refers to a reduction in wound healing, skin barrier function, epidermal water retention, sweat secretion, sebum secretion, vitamin D production, physical damage protection, chemical abilities, immune response, sensory function, and body temperature control. The kisspeptin or active fragment thereof may be for improving skin aging induced by an exogenous factor or endogenous factor. The extrinsic factor refers to various external factors such as ultraviolet (light), and the endogenous factor is also referred to as a chronological factor, which is mainly caused by the passage of time. Specifically, the skin aging specifically includes not only early symptoms of aging induced by external stimuli such as ultraviolet rays, pollution, smoke, chemicals, etc., but also natural aging phenomena caused by aging of skin cells And is a concept that includes both wrinkles, elasticity reduction, skin burning and drying phenomenon. Also, wrinkles include those in which irritation caused by changes in internal and external factors changes the constituents of the skin tissue to cause wrinkles.

Examples of such skin conditions or conditions associated with the skin include skin aging, skin photoaging, scarring, dermatitis, atopic dermatitis, pruritus, eczematous skin disease, dry eczema, erythema, urticaria, psoriasis, drug rash or acne, , Insect and parasitic mediated diseases, superficial dermatophytosis, bacterial infectious diseases, viral diseases, adult diseases, autoimmune vesicular diseases, connective tissue diseases, dyspepsia, pigmented sclerosis and the like.

The skin damage is caused by external physical damage, penetration of chemicals, bacteria, fungi, viruses, exposure to ultraviolet rays, water loss of skin, wrinkles due to aging, oxidation by free radicals (active oxygen) May include damage from a clinical and cosmetic point of view of the skin tissue or cells including skin inflammation, seborrheic dermatitis, redness (redness, erythema), edema, burning, eczema, pruritus (itching), and atopic dermatitis .

The skin includes all parts of the body including the face, hands, arms, legs, feet, chest, belly, back, buttocks, and scalp.

Examples of conditions associated with the inflammatory disease or inflammation include dermatitis, allergy, atopy, conjunctivitis, periodontitis, rhinitis, otitis, sore throat, tonsillitis, pneumonia, gastric ulcer, gastritis, Crohn's disease, colitis, gout, ankylosing spondylitis, rheumatic fever, lupus, Inflammatory bowel disease, fibromyalgia, psoriatic arthritis, osteoarthritis, rheumatoid arthritis, periarthritis, tendonitis, hay fever, tendinitis, myositis, hepatitis, cystitis, nephritis, sjogren's syndrome, multiple sclerosis and acute and chronic inflammatory diseases have.

The composition may be a cosmetic composition.

In one embodiment, the cosmetic composition may be one for prevention or improvement of skin aging, for skin moisturizing, for strengthening skin barrier, for preventing or improving skin wrinkles, for preventing or improving skin damage, or for preventing or improving skin inflammation .

The cosmetic composition may be, for example, a cosmetic formulation of softening agent, nutritional lotion, massage cream, nutritional cream, essence, pack, gel, ampoule or skin sticking type.

The ingredients contained in the cosmetic composition may contain, in addition to the above-mentioned composition, the components conventionally used in cosmetic compositions, for example, conventional auxiliary agents such as stabilizers, solubilizers, vitamins, . ≪ / RTI >

In addition, the composition may be a composition for external application to the skin.

In the present specification, the external preparation for skin may be a cream, a gel, an ointment, a skin emulsifier, a skin suspension, a transdermal patch, a drug-containing bandage, lotion, or a combination thereof. The external preparation for skin may be used in the form of an ointment, a lotion, a spray, a patch, a cream, a powder, a suspension, a gel or a gel. The external preparation for skin is usually used as a component used in external skin preparations such as cosmetics or medicines such as an aqueous component, an oily component, a powder component, an alcohol, a moisturizer, a thickener, an ultraviolet absorber, a whitening agent, an antiseptic, , Coloring agents, various skin nutrients, or a combination thereof, and may be suitably blended as necessary. The external preparation for skin may be a metal blocker such as sodium edetate, sodium edetate, sodium citrate, sodium polyphosphate, sodium metaphosphate or gluconic acid, caffeine, tannin, bellapamil, licorice extract, glabridine, Vitamin C, ascorbic acid magnesium phosphate, ascorbic acid glucoside, arbutin, kojic acid, glucose, fructose, fructose, fructose and other herbal medicines, various herbal medicines, tocopherol acetate, glycyrrhizic acid, Sugars such as trehalose and the like can also be appropriately compounded.

The composition may also be a quasi-drug composition.

The term "quasi-quasi-drugs" means fibers, rubber products or the like, which are used for the purpose of treating, alleviating, treating or preventing diseases of humans or animals, weak acting on the human body or acting directly on the human body, Products similar to or similar to those used in the manufacture of a medicinal product for use in the prevention or treatment of an infectious disease, for use in the preparation of a medicament for the prevention of infection, for use in the diagnosis, treatment, alleviation, treatment or prevention of a human or animal condition or disease, Machinery, or apparatus, and that is not an apparatus, machine, or apparatus of an article used for the purpose of imparting pharmacological effects to the structure or function of a person or animal. .

When the kisspeptin or active fragment thereof is added to the quasi-drug composition, the geraniol may be added as it is or may be used in combination with other quasi-drugs, and may be suitably used according to a conventional method. The amount of the active ingredient to be mixed can be suitably determined according to the intended use (prevention, health or therapeutic treatment).

The quasi-drug composition includes, but is not limited to, personal hygiene products, external preparation for skin, disinfectant cleaner, shower foam, gagrin, wet tissue, detergent soap, hand wash, humidifier filler, mask, ointment or filter filler. The personal care product may be a soap, a wet tissue, a tissue paper, a shampoo, a toothpaste, a hair care product, an air freshner gel or a cleaning gel.

The composition may be a pharmaceutical composition.

The pharmaceutical composition may additionally comprise a pharmaceutically acceptable diluent or carrier. The diluent may be lactose, corn starch, soybean oil, microcrystalline cellulose, or mannitol, the lubricant may be magnesium stearate, talc, or combinations thereof. The carrier may be an excipient, a disintegrant, a binder, a lubricant, or a combination thereof. The excipient may be microcrystalline cellulose, lactose, low substituted hydroxy cellulose, or a combination thereof. The disintegrant may be carboxymethylcellulose calcium, starch glycolate sodium, calcium monohydrogen phosphate anhydrate, or a combination thereof. The binder may be polyvinylpyrrolidone, low-substituted hydroxypropylcellulose, hydroxypropylcellulose, or combinations thereof. The lubricant may be magnesium stearate, silicon dioxide, talc, or a combination thereof.

The pharmaceutical composition may be formulated into an oral or parenteral dosage form. Oral administration formulations may be granules, powders, solutions, tablets, capsules, dry syrups, or combinations thereof. The parenteral dosage form may be an injectable.

The composition may be a health functional food composition.

The health functional food composition may be used together with kispeptin or an active fragment thereof alone or another food or food ingredient, and may be suitably used according to a conventional method. The amount of the active ingredient to be mixed can be suitably determined according to the intended use (prevention, health or therapeutic treatment). Generally, the composition of the present specification may be added in an amount of not more than 15 parts by weight based on the raw material in the production of food or beverage. There is no particular limitation on the kind of the health functional food. Among the kinds of health functional foods, the beverage composition may contain various flavors or natural carbohydrates as additional components such as ordinary beverages. The natural carbohydrates are sugar saccharides such as monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, xylitol, sorbitol and erythritol. Examples of sweeteners include natural sweeteners such as tau martin and stevia extract, synthetic sweeteners such as saccharin and aspartame, and the like. The health food composition may further comprise at least one selected from the group consisting of nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and salts thereof, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, The carbonating agent used, or a combination thereof. The health functional food composition may also contain natural fruit juice, fruit juice beverage, flesh for the manufacture of vegetable drinks, or combinations thereof.

In addition, another aspect provides a method of preventing, ameliorating, or treating the condition of an individual comprising treating or administering an effective amount of the above composition to a subject in need thereof.

The state or composition of the individual is the same as described above. The subject may be a mammal, such as a person, a cow, a horse, a pig, a dog, a sheep, a goat, or a cat.

According to one aspect, the kisspeptin or active fragment thereof has an inhibitory activity of MMP-1 or an inflammatory cytokine and is useful for preventing, improving or treating skin aging, skin damage, skin condition, or inflammatory disease It is effective.

1 is a graph showing the results of HPLC analysis of active fragments of kisspeptin according to one embodiment.

FIG. 2 is a diagram showing the chemical structure and the three-dimensional predicted structural model for active fragments of kispeptin according to one embodiment; FIG. (A) chemical structure of kisspeptin-1 (C ₆₃ H ₈₃ N ₁₇ O ₁₄ ); (b) Three-dimensional predictive structural model; (C) chemical structure of kisspeptin-E (C ₃₁ H ₃₉ N ₉ O ₉ ); (d) Three-dimensional expected structure.

FIG. 3 is a graph showing that kisspeptin-1 and kispeptin-E according to one embodiment inhibit the expression of MMP-1.

FIG. 4 is a graph showing that kisspeptin-1 and kispeptin-E according to one embodiment inhibit the expression of IL-6.

FIG. 5 is a graph showing that kisspeptin-1 and kispeptin-E according to one embodiment inhibit the expression of IL-8.

FIG. 6 is a graph showing that kisspeptin-1 and kispeptin-E according to one embodiment inhibit the expression of 11? -HSD1.

Hereinafter, the present invention will be described in more detail with reference to Examples. However, these embodiments are intended to illustrate one or more embodiments, and the scope of the present invention is not limited to these embodiments.

Example 1. Preparation of kisspeptin active fragments

Kisspeptin-1 (the 45th to 54th amino acid sequence (SEQ ID NO: 3) from the N-terminus of the amino acid sequence of the entire kispeptin) and SEQ ID NOs: 2 and 4 of the kisspeptin active fragment from the kisspeptin represented by SEQ ID NO: .

[Synthetic Scheme]

First, 1) obtaining NH2-protected amino acid residue-resin by Solid-Phase Peptide Synthesis (SPPS); 2) obtaining a peptide from which a protecting group has been removed using a cleavage solution; 3) purifying the obtained non-purified peptide using HPLC to produce an active fragment of the kisspepty. Specifically, SEQ ID NO: 2 was synthesized by a conventional solid phase peptide synthesis method using 9-fluorenylmethoxycarbonyl (Fmoc) as an amino acid protecting group, and the resin was Fmoc-conjugated with phenyl- Wang resin was used. The extension of the amino acid residues was carried out by using N-hydroxybenzotriazole (HOBt) and N, N'-Diisopropyl carbodiimide (DIC) as active agents. Amino acids, HOBt, and DIC used in each step were reacted for 5 hours at room temperature for 2 hours. After completion of the reaction, the resultant was washed six times with dimethylformamide (DMF), and then washed six times with DCM (dichloromethane), followed by drying of the resin. The dried peptide of SEQ ID NO: 1-Resin was reacted with a mixed solution of trifluoroacetic acid: triisopropylsilane: water (90: 5: 5 (v / v)) at room temperature for 2 hours to isolate the peptide from the resin, The peptide was precipitated with ether and crude peptide was obtained using a centrifuge. The obtained peptide of SEQ ID NO: 1 was purified by reversed phase high performance liquid chromatography (Kromasil, C18, 5 mu, 110 Å) using acetonitrile and water containing 0.1% trifluoroacetic acid as a solvent. 250 * 21.2 mm) (yield: 34.8%). Sequence Nos. 3, 4 and 5 were also synthesized in the same manner as above, and the yields were 32.4%, 38.0%, and 35%, respectively.

Experimental Example 1. Evaluation of MMP-1 Inhibition of Kiss Peptin

In order to confirm that the kisspeptin mimetic peptide has skin aging and skin damaging inhibitory activity, the inhibitory effect of kisspeptin on MMP-1 was evaluated.

Specifically, the kisspeptin is Kisspeptin 1 (SEQ ID NO: 3) capped with NH2 in the 45th to 54th amino acid sequence from the N-terminus of the entire kisspeptin amino acid sequence (SEQ ID NO: 1) and Kisspeptin E (SEQ ID NO: 5) capped with NH2 in the 45th to 49th amino acid sequence from the N-terminus of the amino acid sequence of the peptin (SEQ ID NO: 1)) was used to confirm the inhibition of MMP-1.

In order to compare the inhibitory effect of kisspeptin on collagenase (MMP-1), which is increased by ultraviolet light, the amount of MMP-1 expression was measured using human fibroblast cell line Hs68. Fibroblast cell line Hs68 after a busy as a 6-well plate the number of 4x10 ^5, and incubated for 24 hours at 37 ℃, an incubator of 5% CO ₂ condition. After the medium was removed, DPBS was added and UVB of 12 mJ / cm ² was irradiated to the remaining cells except the non-UVB group. Next, ascorbic acid (AA) as a positive control group, and kisspeptin mimetic peptide as an experimental group were added at different concentrations and further cultured for 24 hours. RNA was isolated from the cells treated with each sample using triazole (RNA iso, DAKARA, Japan), and RNA was quantitated at 260 nm using nanodrop, and 2 μg of RNA (C1000 Thermal Cycler, Bio-Rad, USA). Real-time PCR is performed in a real-time PCR machine using a mixture of a target protein, MMP-1 primer, and a cyanine dye, SYBR Green supermis (Applied Biosystems, USA) And the degree of expression of the MMP-1 gene was finally evaluated. The sequences of the primers for PCR and the reaction conditions are shown in Table 1 below. The expression level of the gene was finally analyzed by correction for the β-actin gene, and the results are shown in FIG.

Primer	order	Reaction conditions
MMP-1 forward primer	SEQ ID NO: 8	After activation of the polymerase at 94 ° C for 5 minutes, polymerization was carried out at 40 ° C for 30 seconds at 95 ° C for 30 seconds, at 55 ° C for 30 seconds, and at 72 ° C for 30 seconds
MMP-1 reverse primer	SEQ ID NO: 9
β-actin forward primer	SEQ ID NO: 10
β-actin reverse primer	SEQ ID NO: 11

1 is a graph showing that kisspeptin according to one embodiment inhibits the expression of MMP-1.

As a result, as shown in Fig. 1, the amount of expression of MMP-1 was rapidly increased in the UVB irradiation group, whereas the expression level of the MMP-1 expression decreased in the kisspeptin-treated group. The above results indicate that kisspeptin inhibits skin aging and skin damage by inhibiting the expression amount of MMP-1.

Experimental Example 2. Evaluation of proinflammatory factors for kispeptin

In order to confirm that kispeptin has anti-inflammatory activity, the inhibition of IL-6 and IL-8, the pro-inflammatory factors of kispeptin, was evaluated.

Specifically, the fibroblast cell line Hs68 the cells were cultured in 6 well plates and then dispensed as the number of 4x10 ^5, in 37 ℃, an incubator of 5% CO ₂ condition for 24 hours. Subsequently, the medium was removed and DPBS was added. After UVB irradiation of 15 mJ / cm ² to the remaining cells except for the UVB non-control group, kisspeptin was added by concentration (1 nM, 10 nM, 100 nM, and 1000 nM) Lt; / RTI > for 24 hours. Then, the expression levels of IL-6 and IL-8 overexpressed by UVB were evaluated in the same manner as in Experimental Example 1, except that the primers shown in Table 2 were used. The results are shown in FIGS. 2 and Respectively.

Primer	order	Reaction conditions
IL-6 forward primer	SEQ ID NO: 12	After activation of the polymerase at 94 ° C for 5 minutes, polymerization was carried out at 40 ° C for 30 seconds at 95 ° C for 30 seconds, at 55 ° C for 30 seconds, and at 72 ° C for 30 seconds
IL-6 reverse primer	SEQ ID NO: 13
IL-8 forward primer	SEQ ID NO: 14
IL-8 reverse primer	SEQ ID NO: 15

Figure 2 is a graph showing that kisspeptin according to one embodiment inhibits the expression of IL-6.

Figure 3 is a graph showing that kisspeptin according to one embodiment inhibits the expression of IL-8.

As shown in FIG. 2 and FIG. 3, the expression of LPS-induced IL-6 and IL-8 in the kispeptin-treated group was significantly reduced as compared with the untreated control group. In particular, the expression level of IL-6 was decreased to a level similar to that of the normal control. The above results indicate that kisspeptin according to one embodiment has an anti-inflammatory activity and can be usefully used as a composition for preventing, ameliorating, or treating inflammation, for example, skin inflammation or skin damage.

EXPERIMENTAL EXAMPLE 3 Evaluation of Inhibitory Effect of Kispeptin on Stress Factor

The increase in the activity and expression of 11? -HSD1 by ultraviolet rays is closely related to skin aging, and thus has been attracting attention as a new target for inhibiting skin aging.

Thus, it was confirmed in this experimental example that the kisspeptin mimetic peptide inhibits mRNA expression of 11? -HSD1 induced by UVB.

Specifically, human fibroblasts were inoculated in a 6-well cell culture dish at a density of 4 × 10 ⁵ , and cultured in a 5% CO ₂ incubator at 37 ° C. for 24 hours. UVB was irradiated at 12 mJ, followed by ascorbic acid as a positive control group and kisspeptin mimetic peptide as an experimental group, followed by culturing for 24 hours. The cells were then recovered and RNA was isolated by adding 1 ml of trizol (RNA iso, DAKARA, Japan). Then, the expression level of 11? -HSD1 overexpressed by UVB was evaluated in the same manner as in Experimental Example 1, except that the primer shown in Table 3 below was used. The results are shown in FIG.

primer	order	Reaction conditions
11? -HSD1 forward primer	SEQ ID NO: 16	Polymerase activation at 94 ° C for 5 minutes, polymerization at 95 ° C for 30 seconds, 50 ° C for 1 minute, and 72 ° C for 1 minute 40 cycles
11 [beta] -HSD1 reverse primer	SEQ ID NO: 17

FIG. 4 is a graph showing the inhibition of 11? -HSD1 expression of kisspeptin according to one embodiment. As shown in FIG. 4, the expression of 11β-HSD1, a stress hormone converting enzyme, in the kispeptin-1 and kispeptin-E treated groups was significantly reduced compared to the untreated control group. Especially, in the case of kispeptin-E, the expression of 11? -HSD1 could be remarkably suppressed compared with that of ascorbic acid, which is a positive control.

The above results indicate that kisspeptin according to one embodiment can effectively inhibit skin aging which can be caused by external stimuli by inhibiting the anti-inflammatory activity as well as skin stress hormone-inducing enzyme expression.

Claims (14)

1. A cosmetic composition comprising kisspeptin or an active fragment thereof as an active ingredient.
2. The cosmetic composition according to claim 1, wherein the kispeptin is an amino acid sequence of SEQ ID NO: 1.
3. The cosmetic composition according to claim 1, wherein the active fragment of kispeptin comprises any one of amino acid sequences selected from the group consisting of SEQ ID NOS: 2-7.
4. The cosmetic composition according to claim 1, which is for prevention or improvement of skin aging, for skin moisturizing, for strengthening skin barrier, for preventing or improving skin wrinkles, for preventing or improving skin damage, or for preventing or improving skin inflammation.
5. The cosmetic composition according to claim 1, which is for preventing or improving any one selected from the group consisting of skin wounds, dermatitis, atopic dermatitis, pruritus, eczematous skin disease, dry eczema, erythema, urticaria, psoriasis, .
6. The cosmetic composition according to claim 1, wherein the kisspeptin or an active fragment thereof inhibits MMP (matrix metalloproteinase) or an inflammatory factor.
7. A composition for external application for skin for preventing, improving or treating a skin condition comprising kispeptin or an active fragment thereof as an active ingredient.
8. The skin external preparation according to claim 6, wherein the skin condition is any one or more selected from the group consisting of skin wounds, dermatitis, atopic dermatitis, pruritus, eczematous skin disease, dry eczema, erythema, urticaria, psoriasis, Composition.
9. A composition for external application for skin for preventing or ameliorating skin aging comprising kispeptin or an active fragment thereof as an active ingredient.
10. A pharmaceutical composition for the prevention and treatment of inflammatory diseases comprising kispeptin or an active fragment thereof as an active ingredient.
11. The method of claim 10, wherein the inflammatory disease is selected from the group consisting of dermatitis, allergies, atopy, conjunctivitis, periodontitis, rhinitis, otitis, sore throat, tonsillitis, pneumonia, gastric ulcer, gastritis, Crohn's disease, colitis, arthritis, ankylosing spondylitis, Myositis, hepatitis, cystitis, and nephritis. ≪ RTI ID = 0.0 > 18. < / RTI >
12. A health functional food for improving skin condition comprising kispeptin or an active fragment thereof.
13. A method of preventing, ameliorating or treating a skin condition comprising administering kisspeptin or an active fragment thereof to a subject in need thereof.
14. Use of kispeptin or active fragments thereof for use in the manufacture of a composition for the prevention, amelioration or treatment of a skin condition.

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