A new composition containing catnip-derived exosomes as an active ingredient

Abstract

The present invention provides a composition for improving skin elasticity, improving skin wrinkles, skin regeneration, anti-inflammation, wound healing or promoting wound healing, comprising catnip-derived exosomes as an active ingredient. The composition of the present invention is less likely to contain residual solvents and other impurities compared to conventional catnip hot water extracts, catnip solvent extracts and filtrates of such extracts, and has excellent effects of improving skin elasticity, improving skin wrinkles, skin regeneration, anti-inflammation, wound healing or promoting wound healing.

Description

[Technical Field]

[0001] The present invention relates to a new composition comprising exosomes derived from catnip (Nepeta cataria) as an active ingredient, and more particularly to a composition for improving skin elasticity, alleviating skin wrinkles, skin regeneration, anti-inflammatory properties, wound healing, or promoting wound healing, comprising exosomes derived from catnip as an active ingredient.

[0002] The present invention also relates to a cosmetic composition for improving skin elasticity, reducing skin wrinkles, or regenerating skin, and a pharmaceutical composition for anti-inflammation, wound healing, or promoting wound healing, which contains the composition. [Background technology]

[0003] As skin ages, skin elasticity decreases and wrinkles increase. This is known to occur due to a decrease in collagen synthesis and an increase in the expression of matrix metalloproteinase (MMP), an enzyme that breaks down collagen.

[0004] Additionally, aging and UV exposure are known to increase the synthesis of prostaglandin E2 in skin cells due to an increase in COX-2, an enzyme that produces inflammatory cytokines, leading to increased production of pro-inflammatory factors. Inflammatory responses increase MMP biosynthesis, leading to collagen degradation, resulting in decreased skin elasticity and the formation of wrinkles. Direct exposure of the skin to sunlight or UV rays generates a large amount of free radicals, which damage the skin's antioxidant defense system, increasing wrinkles and causing skin laxity, accelerating skin aging. Substances known to be effective in reducing wrinkles include adenosine and retinoic acid. However, adenosine has limited clinical efficacy, and retinoic acid cannot be used in pregnant women and has side effects such as erythema.

[0005] Inflammation is a defense mechanism that occurs in response to pathological conditions caused by physical or chemical trauma, bacterial, fungal, or viral infections, and various allergens. It is a part of the innate immune response. Various substances and physiological and chemical phenomena are involved in the inflammatory response, but recent research has revealed that various inflammatory cytokines play an important role in the inflammatory response. Major cytokines involved in the inflammatory response include IL-1β, TNF-α, IL-6, IL-8, IL-12, and IFN-β. Increased expression and secretion of these cytokines, along with their activation, are associated with a series of complex physiological responses, including the secretion of inflammatory mediators, immune cell infiltration, cell migration, and tissue destruction, as well as symptoms such as erythema, edema, fever, and pain.

[0006] Generally, inflammatory responses are not a major problem if the source of infection is removed and damaged tissue is regenerated, and the affected area will recover normally. However, if the source of infection is not removed or if the inflammatory response is excessive or persistent due to internal substances, it can lead to acute or chronic inflammatory diseases. Nonsteroidal anti-inflammatory drugs, steroidal anti-inflammatory drugs, neuropeptide antagonists, COX inhibitors, antihistamines, and immunosuppressants such as cyclosporin A are used to alleviate or treat inflammatory responses and resulting inflammatory diseases. However, these drugs have the drawback of causing side effects such as skin atrophy, vasodilation, depigmentation, hypersensitivity reactions, tolerance, and neutropenia. Furthermore, these drugs are limited in that they only help to control symptoms to an appropriate level rather than providing a fundamental treatment.

[0007] Meanwhile, recent research has shown that cell secretomes contain various bioactive factors that regulate cell behavior. In particular, cell secretomes contain exosomes, which have intercellular signaling functions, and research into their components and functions is actively underway.

[0008] Cells release various membrane-type vesicles into the extracellular environment; these vesicles are commonly referred to as extracellular vesicles (EVs). EVs are also known as plasma membrane-derived vesicles, ectosomes, shedding vesicles, microparticles, and exosomes, and are sometimes referred to separately from exosomes.

[0009] Exosomes are endoplasmic reticulum measuring tens to hundreds of nanometers in size and possessing a double phospholipid membrane similar to the cell membrane, which contains exosomal cargo, including proteins and nucleic acids (e.g., mRNA and miRNA). Exosome cargo contains a wide range of signaling factors, which are known to be cell-type specific and differentially regulated depending on the environment of the secreting cell. Exosomes are intercellular signaling mediators secreted by cells, and the various cellular signals transmitted by them are known to regulate target cell behaviors, including activation, growth, migration, differentiation, dedifferentiation, apoptosis, and necrosis. Exosomes contain specific genetic material and bioactive factors depending on the nature and state of the cells from which they are derived. Exosomes derived from proliferating stem cells regulate cell behaviors, such as cell migration, proliferation, and differentiation, reflecting the stem cell properties related to tissue regeneration (Non-Patent Document 1).

[0010] In other words, exosomes, which are called cellular avatars, contain bioactive factors such as growth factors just like cells, but act as carriers that carry bioactive factors between cells, i.e., as a means of communication between cells. Exosomes are known to be released not only from animal cells such as stem cells, immune cells, fibroblasts, and cancer cells, but also from cells of various organisms, including plants, bacteria, fungi, and algae. For example, exosomes can be isolated from the culture medium of cancer cells, immune cells, mesenchymal stem cells, and the like, as well as from the culture medium of plant cells or plant stem cells.

[0011] However, research into the isolation, purification, and characterization of exosomes derived from plant cells or plant stem cells is still in its infancy, and most of these studies simply refer to extracellular vesicles present in the filtrate of plant juice as exosomes for marketing purposes. Therefore, more detailed characterization and functional studies of plant cell-derived exosomes are needed.

[0012] Catnip, scientifically known as Nepeta cataria, is a perennial herb belonging to the Lamiales, Lamiaceae, and Nepeta genus. Nepeta cataria is commonly called catnip because cats love it. Catnip is a type of herb that, when dried, releases a peppermint scent and is also known as catnip or catmint. Catnip is native to Asia and Europe, and is found in Korea, Japan, China, Central Asia, Africa, Europe, and North America. It is known for its pain-relieving and fever-reducing properties. However, current technology is limited to the use of extracts obtained by hot water extraction or solvent extraction of catnip leaves, stems, or the whole plant as ingredients in food and cosmetics. However, solvent extracts pose a risk of human toxicity due to residual solvent residues.

[0013] The present inventors have confirmed that catnip-derived exosomes are effective in improving skin elasticity, reducing skin wrinkles, skin regeneration, anti-inflammation, wound healing or promoting wound healing, etc., and have developed a cosmetic composition for improving skin elasticity, reducing skin wrinkles or skin regeneration, and a pharmaceutical composition for anti-inflammation, wound healing or promoting wound healing, which contain catnip-derived exosomes as an active ingredient.

[0014] On the other hand, it should be understood that the matters described above as background art are merely intended to facilitate understanding of the background of the present invention, and are not cited as an admission that they can be used as "prior art" for the present invention. [Prior art documents] [Non-patent literature]

[0015] [Non-Patent Document 1] Nature Review Immunology 2002(2)569-579 Summary of the Invention [Problem to be solved by the invention]

[0016] An object of the present invention is to provide a composition for improving skin elasticity, reducing skin wrinkles, regenerating skin, anti-inflammatory, wound healing, or promoting wound healing, which comprises exosomes derived from catnip (Nepeta cataria) as an active ingredient.

[0017] Another object of the present invention is to provide a cosmetic composition for improving skin elasticity, reducing skin wrinkles or regenerating skin, and a pharmaceutical composition for anti-inflammation, wound healing or promoting wound healing, which comprises the composition.

[0018] However, the above-mentioned problems of the present invention are merely examples and are not intended to limit the scope of the present invention. Other objects and advantages of the present invention will become clearer from the following detailed description of the invention, the claims, and the drawings. [Means for solving the problem]

[0019] The present invention provides a composition for improving skin elasticity, reducing skin wrinkles, regenerating skin, anti-inflammation, wound healing, or promoting wound healing, which comprises exosomes derived from catnip (Nepeta cataria) as an active ingredient.

[0020] The term "catnip" used herein refers to a perennial plant belonging to the Lamiales, Lamiaceae family, and Nepeta genus, and its scientific name is Nepeta cataria. Catnip is a type of herb that, when dried, gives off a peppermint scent and is also known as catnip or catmint. Catnip is native to Asia and Europe, and is found in Korea, Japan, China, Central Asia, Africa, Europe, North America, etc.

[0021] As used herein, the term "exosomes" refers to nano-sized vesicles having a membrane structure that are secreted or released from plant cells into the extracellular space, and are also defined as exosome-like vesicles or exosome-like particles.

[0022] The term "skin elasticity" as used herein refers to the property of skin that is easily restored to its original shape when deformed by an external force is removed. "Skin wrinkles" refer to fine wrinkles that occur due to skin aging, and can be induced by genetic factors, a decrease in collagen and elastin in the dermis of the skin, the external environment, etc. Therefore, the term "skin wrinkle improvement" as used herein refers to suppressing or inhibiting the formation of wrinkles on the skin or alleviating wrinkles that have already formed.

[0023] As used herein, the term "anti-inflammatory" means preventing, suppressing, alleviating, ameliorating, or treating inflammation, and examples of inflammatory diseases include, but are not limited to, dermatitis, atopic dermatitis, eczema, inflammation caused by bacterial infection, viral infection, or fungal infection, burns, inflammation caused by burns, wounds, and inflammation caused by wounds.

[0024] As used herein, the term "wound" refers to an injury to a living body, and includes pathological conditions in which tissues constituting the internal or external surface of a living body, such as skin, muscle, nerve tissue, bone, soft tissue, internal organs, or vascular tissue, are disrupted or destroyed. Non-limiting examples of wounds include abrasions, lacerations, puncture wounds, cuts, avulsions, bed sores, pressure sores, tissue destruction by radiation, penetrated wounds, gunshot wounds, burns, frostbite, surgical wounds, suture sites after plastic surgery, and chemical wounds, and may include injury to any part of an individual.

[0025] As used herein, the term "iontophoresis" refers to a method of passing a microcurrent through the skin to which an active ingredient has been applied, creating a potential difference and altering the electrical environment of the skin, thereby allowing the ionized active ingredient to permeate the skin through electrical repulsion. Iontophoresis used in one embodiment of the present invention may include a method in which a current from an external power source flows into an electrode patch on the skin, thereby introducing a microcurrent into the skin; a method in which a battery is attached to the electrode patch itself, thereby introducing a microcurrent into the skin; or a method in which a microcurrent is introduced into the skin through a patch equipped with reversed electrodialysis means that generates a current by utilizing the difference in ion concentration between a high-concentration electrolyte solution and a low-concentration electrolyte solution. However, the present invention is not limited thereto, and various types of iontophoresis may be used.

[0026] The term "exosomes derived from catnip (Nepeta cataria)" as used herein is meant to include any exosomes isolated from, or derived from, catnip plant cells or plant stem cells, e.g., secreted and / or released from, e.g., catnip plant cell culture, catnip callus culture, catnip plant stem cell culture, catnip juice, or equivalent catnip biological solutions.

[0027] In one specific example of the composition of the present invention, the catnip-derived exosomes may be those separated and purified from catnip juice.

[0028] In one embodiment of the present invention, a composition comprising catnip (Nepeta cataria)-derived exosomes as an active ingredient may exhibit at least one of the following effects: improving skin elasticity, reducing skin wrinkles, skin regeneration, anti-inflammatory, wound healing, or wound healing promotion.

[0029] The composition of the present invention containing the catnip (Nepeta cataria)-derived exosomes as an active ingredient may be a cosmetic composition or a pharmaceutical composition.

[0030] The present invention provides a cosmetic composition for improving skin elasticity, reducing skin wrinkles, or regenerating skin, comprising exosomes derived from catnip (Nepeta cataria) as an active ingredient. For example, the cosmetic composition may be in the form of a shampoo, soap, rinse, surfactant-containing cleanser, cream, lotion, ointment, tonic, treatment, conditioner, suspension, emulsion, paste, gel, oil, wax, spray, aerosol, mist, or powder, and is preferably a lotion or cream.

[0031] The present invention also provides a pharmaceutical composition for anti-inflammation, wound healing, or wound healing promotion, comprising exosomes derived from catnip (Nepeta cataria) as an active ingredient.

[0032] As a non-limiting example, the pharmaceutical composition of one embodiment of the present invention can be administered or administered by injection, microneedling, iontophoresis, application, or a combination thereof. For example, the pharmaceutical composition may be in the form of an injection, infusion, spray, liquid, or patch.

[0033] When the composition of an embodiment of the present invention is used as a pharmaceutical composition, it may contain a pharmaceutically acceptable carrier, excipient, or diluent. Examples of the carrier, excipient, and diluent include, but are not limited to, lactose, dextrose, trehalose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium carbonate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil. Furthermore, an effective amount of the pharmaceutical composition of an embodiment of the present invention refers to the amount required for administration to achieve anti-inflammatory, wound healing, or wound healing promotion effects.

[0034] The compounding ratio of the pharmaceutical composition of one embodiment of the present invention can be appropriately selected depending on the type, amount, and form of the additional ingredients as described above. For example, the pharmaceutical composition of the present invention may be contained in an amount of about 0.1 to 99% by weight, preferably about 10 to 90% by weight, of the total volume of the injection. Furthermore, the appropriate dosage of the pharmaceutical composition of one embodiment of the present invention can be adjusted depending on the severity of the disease, the type of dosage form, the formulation method, the patient's age, sex, weight, health condition, diet, excretion rate, and the administration time and method. For example, when the pharmaceutical composition of one embodiment of the present invention is administered to an adult, it can be administered at a dose of 0.001 mg / kg to 100 mg / kg per day, in one or several divided doses.

[0035] On the other hand, when the composition of one embodiment of the present invention is produced as a cosmetic composition, components that are usually used in cosmetic compositions, such as moisturizers, antioxidants, oily components, ultraviolet absorbers, emulsifiers, surfactants, thickeners, alcohols, powder components, coloring materials, aqueous components, water, various skin nutrients, etc., can be appropriately blended as needed, within a range that does not impair the effects of the present invention.

[0036] Furthermore, in addition to the exosomes derived from catnip (Nepeta cataria), the cosmetic composition of one embodiment of the present invention may be mixed with a conventional skin improving agent and / or moisturizing agent to the extent that the effects of the exosomes (e.g., improvement of skin condition, improvement of skin elasticity, improvement of skin wrinkles, skin regeneration, etc.) are not impaired.

[0037] The cosmetic composition according to one embodiment of the present invention can be applied in various forms, such as a patch, a mask pack, a sheet mask, a cream, a tonic, an ointment, a suspension, an emulsion, a paste, a lotion, a gel, an oil, a pack, a spray, an aerosol, a mist, a foundation, a powder, or oil paper.

[0038] The cosmetic composition according to one embodiment of the present invention can be used for purposes such as improving skin elasticity, reducing skin wrinkles, and skin regeneration, and can be formulated into any cosmetic formulation commonly used in the art, such as, but not limited to, patches, mask packs, sheet masks, softening lotions, nourishing lotions, astringent lotions, nourishing creams, massage creams, eye creams, cleansing creams, essences, eye essences, cleansing lotions, cleansing foams, cleansing waters, sunscreens, lipsticks, soaps, shampoos, surfactant-containing cleansers, bath additives, body lotions, body creams, body oils, body essences, body washes, hair dyes, and hair tonics.

[0039] The cosmetic composition according to one embodiment of the present invention may contain ingredients commonly used in cosmetic compositions, such as common adjuvants and carriers, such as antioxidants, stabilizers, solubilizers, vitamins, pigments, and fragrances. In addition, in each formulation of the cosmetic composition, other ingredients can be easily selected and blended by those skilled in the art depending on the type of cosmetic composition or the intended use.

[0040] Another embodiment of the present invention provides a cosmetic method for regulating the condition of mammalian skin (excluding therapeutic use) using the cosmetic composition. In the cosmetic method of the present invention, regulating skin condition means improving / causing skin condition to improve or preventatively regulating skin condition, and improving skin condition means a visually and / or tactilely perceptible positive change in the appearance and feel of the skin. For example, improving skin condition may be reducing wrinkles, improving skin elasticity, or regenerating skin.

[0041] A cosmetic method according to one embodiment of the present invention includes (a) directly applying the cosmetic composition to the skin of a mammal, (b) contacting or attaching a patch, mask pack, or sheet mask coated or soaked in the cosmetic composition to the skin of a mammal, or sequentially performing steps (a) and (b). In step (a), the cosmetic composition may be a lotion or a cream.

[0042] Furthermore, the cosmetic method according to one embodiment of the present invention may further include, after step (b), the step of (c) removing the patch, mask pack, or sheet mask from the mammalian skin and applying the cosmetic composition to the mammalian skin. In step (c), a lotion or cream may be used as the cosmetic composition.

[0043] As a non-limiting example of the cosmetic method of one embodiment of the present invention, the cosmetic composition may be applied to the skin by microneedling, iontophoresis, direct application, or a combination thereof. For example, the cosmetic composition may be in the form of a spray, liquid, or patch.

[0044] In one embodiment of the cosmetic method of the present invention, the mammal may be a cat, human, dog, rodent, horse, cow, monkey, or pig.

[0045] The present invention also provides a method for treating inflammation, treating wounds, or accelerating wound healing, comprising the step of administering a therapeutically effective amount of the pharmaceutical composition to a mammal, or applying the pharmaceutical composition to the skin, an inflammation site, or a wound site.

[0046] As a non-limiting example, in a method for treating inflammation, treating a wound, or promoting wound healing according to one embodiment of the present invention, the pharmaceutical composition may be administered by injection, microneedling, iontophoresis, or a combination thereof. For example, the pharmaceutical composition may be in the form of an injection, infusion, spray, liquid, or patch.

[0047] In one embodiment of the method for treating inflammation, treating wounds or promoting wound healing of the present invention, the mammal may be a cat, a human, a dog, a rodent, a horse, a cow, a monkey, or a pig. [Effects of the Invention]

[0048] The composition of the present invention is less likely to contain residual solvents and other impurities than conventional catnip hot water extracts, catnip solvent extracts, and filtrates of such extracts, and has excellent effects of improving skin elasticity, reducing skin wrinkles, skin regeneration, anti-inflammatory properties, and wound healing or wound healing promotion.

[0049] On the other hand, the scope of the present invention is not limited by the effects described above. [Brief explanation of the drawings]

[0050] [Figure 1] 1 is a graph showing particle size distribution and particle number obtained by performing NTA (nanoparticle tracking analysis) on catnip-derived exosomes of the present invention. [Figure 2] These are cell fluorescence microscope images confirming that fluorescently stained catnip-derived exosomes were delivered to human skin fibroblasts (green: exosomes delivered into cells, blue: cell nuclei). [Figure 3] 1 is a graph showing the exosome concentration-dependent increase in the relative collagen amount when human skin fibroblasts are treated with catnip-derived exosomes. [Figure 4] 1 is a graph showing increased migration of human dermal fibroblasts after treatment of Scratch-Wound with catnip-derived exosomes. [Figure 5] This is a cell fluorescence microscope image confirming that fluorescently stained catnip-derived exosomes were delivered to RAW264.7 cells (green: exosomes delivered into cells, blue: cell nuclei). [Figure 6] 10 is a graph showing that treatment of RAW264.7 cells with catnip-derived exosomes reduces LPS-induced IL-6. DETAILED DESCRIPTION OF THE INVENTION

[0051] The present invention will be described in more detail with reference to the following examples. However, the following examples are merely illustrative of the contents of the present invention and are not intended to limit or restrict the scope of the present invention. Anything that can be easily inferred by a person skilled in the art from the detailed description and examples of the present invention is considered to be within the scope of the present invention. The references cited in this specification are incorporated herein by reference.

[0052] Throughout the specification, when a part is said to "comprise" certain elements, this does not mean that it excludes other elements, but that it may further include other elements, unless specifically stated to the contrary. [Example]

[0053] Example 1: Preparation of catnip juice Catnip (Nepeta cataria) leaves and stems were first washed with running water to remove pesticides. Next, the catnip leaves and stems were cut into lengths of 5 cm or less and squeezed using a slow-speed screw at 40 rpm to obtain a liquid juice, which was then filtered through a 400-mesh mesh to remove large floating particles. The filtered juice was centrifuged at 10,000 × g for 10 minutes at 4°C to remove proteins and cell walls. The supernatant obtained from this process was filtered through a 0.22 μm pore filter membrane to obtain a filtrate, which was stored at -80°C until further purification.

[0054] Example 2: Purification of exosomes derived from catnip Catnip juice obtained as in Example 1 was subjected to centrifugation and ultracentrifugation in stages to remove numerous impurities and isolate catnip-derived exosomes. The thawed catnip juice was centrifuged at 2,000 × g for 10 minutes and then filtered through a 0.22 μm pore membrane to obtain a filtrate. The supernatant was then subjected to a primary centrifugation at 10,000 × g and a secondary centrifugation at 20,000 × g. The supernatant was then collected and ultracentrifuged at 100,000 × g for 70 minutes at 4°C to obtain a pellet. The resulting pellet was suspended in phosphate buffered saline (PBS) to obtain the final catnip-derived exosomes. The size and concentration of the catnip-derived exosomes isolated by this method were confirmed by nanoparticle tracking analysis (NTA) using an NS300 (purchased from Malvern Panalytical) (Figure 1).

[0055] Example 3: Confirmation of the transduction ability of catnip-derived exosomes to skin fibroblasts To confirm whether catnip-derived exosomes were delivered into human dermal fibroblasts (purchased from ATCC), the following analysis was performed. The membranes of the catnip-derived exosomes prepared in Example 2 were fluorescently stained by reacting them with PKH67 fluorescent dye (purchased from Sigma-Aldrich). After the reaction, the reaction solution was fractionated on a MiniTrap-25 column (purchased from Cytiva) to remove free PKH67 fluorescent dye that did not stain the exosome membrane. For the negative control group, PKH67 fluorescent dye was reacted with a buffer solution and then fractionated on a MiniTrap-25 column. The PKH67-stained exosomes were then cultured with pre-prepared human dermal fibroblasts, and the time course of exosome delivery into the cells was observed using a fluorescence microscope. Hoechst fluorescent dye (purchased from Thermo Fisher Scientific) was used to stain the cell nuclei, and CellMask Orange fluorescent dye (purchased from Thermo Fisher Scientific) was used to stain the cytoplasm. To confirm whether exosomes were delivered into cells, we confirmed that fluorescently stained exosomes were delivered into cells, and that green fluorescence accumulated inside the cells over time (Figure 2).

[0056] Example 4: Confirmation of collagen production promoting effect Human dermal fibroblasts (purchased from ATCC) dispersed in DMEM medium containing fetal bovine serum were dispensed into multiwell plates and cultured for 24 hours, followed by an additional 48 hours of culture in serum-free medium. The catnip-derived exosomes prepared in Example 2 were then diluted in serum-free medium and applied to the human dermal fibroblasts, which were then cultured. To confirm the efficacy of collagen production using human dermal fibroblasts, experimental groups were divided as follows: (1) Negative control group (Control): Experimental group treated with serum-free medium only; (2) Catnip-derived exosome low concentration treatment group (shown as "low concentration" in Figure 3): An experimental group in which catnip-derived exosomes prepared in Example 2 were diluted in serum-free medium and treated (treatment concentration: 1.0 × 10 10 particles / mL); (3) Catnip-derived exosome high concentration treatment group (shown as "high concentration" in Figure 3): an experimental group in which catnip-derived exosomes prepared in Example 2 were diluted in serum-free medium and treated (treatment concentration: 4.0 × 10 10 particles / mL).

[0057] Human dermal fibroblasts in each experimental group were cultured for 24 hours after treatment, and the culture medium was collected and centrifuged to prepare the centrifuged culture medium. The amount of collagen synthesized by human dermal fibroblasts and accumulated in the culture medium was measured using an EIA kit for procollagen type IC peptide (PIP) (purchased from Takara). The measured collagen amount was normalized by dividing it by the total cell number measured using an MTT assay kit (purchased from Sigma-Aldrich) to determine the relative collagen content.

[0058] As a result, it was confirmed that the catnip-derived exosomes of the present invention significantly increased collagen synthesis in human skin fibroblasts in a concentration-dependent manner compared to the negative control group (Figure 3).

[0059] From the above experimental results, it can be seen that the catnip-derived exosomes of the present invention have excellent collagen synthesis increasing efficacy, i.e., skin elasticity improvement, wrinkle reduction and / or skin regeneration efficacy.

[0060] These results demonstrate that the catnip-derived exosomes of the present invention have functional activity, i.e., the activity of increasing collagen synthesis, which is useful for cosmetics for improving skin elasticity, reducing wrinkles, and / or regenerating skin. Therefore, the catnip-derived exosomes of the present invention can be usefully used as an active ingredient in cosmetic compositions for improving skin elasticity, reducing wrinkles, and / or regenerating skin.

[0061] <Example 5: Confirmation of skin regeneration efficacy using skin fibroblasts> A scratch-wound assay was performed to evaluate whether catnip-derived exosomes prepared as in Example 2 promoted wound healing in human dermal fibroblasts (Human Dermal Fibroblasts; purchased from ATCC). Human dermal fibroblasts dispersed in DMEM medium containing fetal bovine serum were seeded into a wound-inducing culture plate (ImageLock Plate; purchased from EssenBio) at a density of 5,000 cells / well and cultured at 5% CO2 and 37°C for 24 hours until they reached 90% or greater confluency. Scratches were then induced using a WoundMaker (EssenBio). To assess the efficacy of skin regeneration using human dermal fibroblasts, experimental groups were divided as follows: (1) Negative control group (NC): Experimental group treated with serum-free medium only; (2) Positive control group (PC): an experimental group treated with culture medium containing 10% fetal bovine serum; (3) Catnip-derived exosome low concentration treatment group (shown as "low concentration" in FIG. 4): an experimental group in which catnip-derived exosomes prepared in Example 2 were diluted in serum-free medium and treated (treatment concentration: 2.0 × 10 9 particles / mL); (4) Catnip-derived exosome high concentration treatment group (shown as "high concentration" in FIG. 4): an experimental group in which catnip-derived exosomes prepared in Example 2 were diluted in serum-free medium and treated (treatment concentration: 1.2 × 10 10 particles / mL).

[0062] Then, each experimental group was treated with a scratch wound and cultured with human dermal fibroblasts at 37°C under 5% CO2 for 12 hours, and the wound healing ability was measured using Incucyte (purchased from Sartorius).

[0063] As a result of measuring wound healing ability, it was confirmed that the catnip-derived exosomes of the present invention increased the migration of human skin fibroblasts compared to the negative control group (Figure 4).

[0064] The above experimental results demonstrate that the catnip-derived exosomes of the present invention have excellent efficacy in promoting the migration of human skin fibroblasts, i.e., excellent wound healing or skin regeneration efficacy.

[0065] Therefore, the catnip-derived exosomes of the present invention can be usefully used as an active ingredient in a cosmetic composition for improving skin elasticity, reducing skin wrinkles and / or regenerating skin, and a pharmaceutical composition for treating wounds or promoting wound healing.

[0066] Example 6: Confirmation of macrophage transduction ability of catnip-derived exosomes To confirm whether catnip-derived exosomes were delivered into mouse macrophages (RAW264.7; purchased from ATCC), the following analysis was performed. The membranes of catnip-derived exosomes prepared in Example 2 were fluorescently stained by reacting them with PKH67 fluorescent dye (purchased from Sigma-Aldrich). After the reaction, the reaction solution was fractionated using a MiniTrap-25 column (purchased from Cytiva) to remove free PKH67 fluorescent dye that did not stain the exosome membrane. As a negative control, PKH67 fluorescent dye was reacted with a buffer solution and then fractionated using a MiniTrap-25 column. The PKH67-stained exosomes were cultured with pre-cultured mouse macrophages, and then intracellular delivery of the exosomes was observed over time using a fluorescence microscope. Hoechst fluorescent dye (purchased from Thermo Fisher Scientific) was used to stain the cell nuclei, and CellMask Orange fluorescent dye (purchased from Thermo Fisher Scientific) was used to stain the cytoplasm. The results confirmed that the fluorescently stained exosomes were delivered into the cells, and that green fluorescence accumulated within the cells over time (Figure 5).

[0067] Example 7: Evaluation of the anti-inflammatory efficacy of catnip-derived exosomes To confirm whether the catnip-derived exosomes prepared as in Example 2 exhibit anti-inflammatory effects, we examined the effect of the catnip-derived exosomes on IL-6 production in RAW264.7 cells, a mouse macrophage. RAW264.7 cells were suspended in DMEM medium containing 10% FBS and dispensed into each well of a multiwell plate to achieve 80-90% confluency. The next day, catnip-derived exosomes diluted in fresh LPS-containing medium (DMEM medium containing 1% FBS and 200 nM LPS (lipopolysaccharide)) were treated with RAW264.7 cells, and the RAW264.7 cells were then cultured for 24 hours. Experimental groups for evaluating anti-inflammatory effects were divided as follows: (1) Negative control group (NC): RAW264.7 cells were treated with LPS medium alone; (2) Positive control (PC): RAW264.7 cells were treated with LPS medium containing dexamethasone (final concentration: 200 μM); (3) Catnip-derived exosome low concentration treatment group (shown as "low concentration" in FIG. 6): An experimental group in which the catnip-derived exosomes prepared in Example 2 were diluted in LPS medium and treated with RAW264.7 cells (treatment concentration: 6.0 × 10 9 particles / mL); (4) Catnip-derived exosome high concentration treatment group (shown as "high concentration" in FIG. 6): An experimental group in which the catnip-derived exosomes prepared in Example 2 were diluted in LPS medium and treated with RAW264.7 cells (treatment concentration: 2.0 × 10 10 particles / mL).

[0068] After the culture was completed, the culture supernatant was collected and the amount of IL-6, an inflammatory cytokine present in the culture supernatant, was measured using an IL-6 ELISA kit. Following the manufacturer's instructions, the amount of IL-6 (an inflammatory cytokine) produced in the group treated with LPS alone and in the experimental groups treated with dexamethasone and catnip-derived exosomes (low and high concentrations) mixed with LPS medium was confirmed (Figure 6).

[0069] As a result, it was confirmed that the production of IL-6 was suppressed in the experimental groups in which RAW264.7 cells were treated with LPS and the catnip-derived exosomes (low and high concentrations) prepared in Example 2, compared to the negative control group (Figure 6).

[0070] The above experimental results demonstrate that the catnip-derived exosomes of the present invention have excellent anti-inflammatory effects. Therefore, the catnip-derived exosomes of the present invention can be usefully used as an active ingredient in pharmaceutical compositions for anti-inflammatory, wound healing, and / or wound healing promotion.

[0071] Although the present invention has been described with reference to the above examples, the present invention is not limited thereto. Those skilled in the art will recognize that modifications and variations can be made without departing from the spirit and scope of the present invention, and that such modifications and variations are also included within the scope of the present invention.

Claims

1. A cosmetic composition for improving skin elasticity, improving skin wrinkles, or skin regeneration, comprising exosomes derived from catnip (Nepeta cataria) as an active ingredient.

2. 2. The cosmetic composition for improving skin elasticity, improving skin wrinkles or regenerating skin according to claim 1, which is in the form of a shampoo, soap, rinse, surfactant-containing cleanser, cream, lotion, ointment, tonic, treatment, conditioner, suspension, emulsion, paste, gel, oil, wax, spray, aerosol, mist or powder.

3. A pharmaceutical composition for anti-inflammation, wound healing or wound healing promotion, comprising exosomes derived from catnip (Nepeta cataria) as an active ingredient.

4. The pharmaceutical composition for anti-inflammatory, wound healing or wound healing promotion according to claim 3, which is administered or treated by injection, microneedling, iontophoresis, application or a combination thereof.

5. 4. The pharmaceutical composition for anti-inflammation, wound healing or wound healing promotion according to claim 3, which is in the form of an injection, infusion, spray, liquid or patch.

6. A cosmetic method for improving the elasticity of mammalian skin, alleviating skin wrinkles, or regenerating skin, excluding therapeutic use, using a cosmetic composition containing exosomes derived from catnip (Nepeta cataria) as an active ingredient.

7. 7. The cosmetic method according to claim 6, comprising: (a) directly applying the cosmetic composition to the skin of a mammal; (b) contacting or attaching a patch, mask pack, or sheet mask to which the cosmetic composition has been applied or soaked, to the skin of a mammal; or sequentially carrying out (a) and (b).

8. The cosmetic method according to claim 7, wherein a lotion or cream is used as the cosmetic composition in step (a).

9. The cosmetic method according to claim 7 or 8, further comprising the step of: (c) after step (b), removing the patch, mask pack, or sheet mask from the mammalian skin, and applying the cosmetic composition to the mammalian skin.

10. The cosmetic method according to claim 9, wherein a lotion or cream is used as the cosmetic composition in step (c).

11. The cosmetic method according to any one of claims 6 to 8, wherein the mammal is a cat, a human, a dog, a rodent, a horse, a cow, a monkey, or a pig.

12. A method for treating inflammation, treating wounds or promoting wound healing, comprising the step of administering a therapeutically effective amount of a pharmaceutical composition comprising exosomes derived from catnip (Nepeta cataria) as an active ingredient to a mammal other than humans, or applying a pharmaceutical composition comprising exosomes derived from catnip (Nepeta cataria) as an active ingredient to the skin, an inflammation site or a wound site of a mammal other than humans.

13. 13. The method for treating inflammation, treating wounds or promoting wound healing according to claim 12, wherein the pharmaceutical composition is administered by injection, microneedling, iontophoresis, or a combination thereof.

14. The method for treating inflammation, treating wounds or promoting wound healing according to claim 12, wherein the pharmaceutical composition is in the form of an injection, infusion, spray, liquid or patch.

15. The method for treating inflammation, treating wounds or promoting wound healing according to any one of claims 12 to 14, wherein the mammal is a cat, dog, rodent, horse, cow, monkey or pig.

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