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, improving skin wrinkles, regenerating skin, anti-inflammation, 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, improving skin wrinkles, or regenerating skin, and a pharmaceutical composition for anti-inflammation, wound healing, or wound healing promotion, which contains the composition. [Background technology]

[0003] As skin aging occurs, skin elasticity decreases and wrinkles increase. It is known that the decrease in skin elasticity and the formation of wrinkles are caused by a decrease in collagen synthesis and an increase in the expression of matrix metalloproteinase (MMP), an enzyme that breaks down collagen.

[0004] In addition, it is known that the synthesis of prostaglandin E2 increases due to the increase in COX-2, an enzyme that produces inflammatory cytokines, in skin cells as a result of aging and exposure to ultraviolet rays, and the production of inflammatory factors increases. Inflammatory reactions increase the synthesis of MMP, which causes collagen decomposition, resulting in a decrease in skin elasticity and the formation of skin wrinkles. In particular, when sunlight or ultraviolet rays are directly irradiated on the skin, a large amount of free radicals are generated, which damage the skin's antioxidant defense system, increasing wrinkles and loosening the skin, accelerating skin aging. Substances known to be effective in improving skin wrinkles include adenosine and retinoic acid, but adenosine has little clinical efficacy, and retinoic acid cannot be used on pregnant women and has side effects such as erythema.

[0005] Inflammation is a defense response of the body in response to pathological conditions caused by physical or chemical trauma, infection with bacteria, fungi or viruses, various allergy-inducing substances, etc. Inflammatory responses are a part of the innate immune response. Various substances and physiological and chemical phenomena are involved in inflammatory responses, and recent studies have revealed that various inflammatory cytokines play an important role in inflammatory responses. The main cytokines involved in inflammatory responses include IL-1β, TNF-α, IL-6, IL-8, IL-12, IFN-β, etc., and the increase in expression and secretion and activation of these cytokines are associated with a series of complex physiological responses such as secretion of inflammatory mediators, infiltration of immune cells, cell migration and tissue destruction, as well as symptoms such as erythema, edema, fever and pain.

[0006] In general, inflammatory reactions are not a big 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 an internal substance causes an excessive or persistent inflammatory reaction, it will lead to acute or chronic inflammatory diseases. To alleviate or treat inflammatory reactions and the resulting inflammatory diseases, immunosuppressants such as nonsteroidal anti-inflammatory drugs, steroidal anti-inflammatory drugs, neuropeptide antagonists, COX inhibitors, antihistamines, and cyclosporine A are used, but they have problems with side effects such as skin atrophy, vasodilation, depigmentation, hypersensitivity reactions, tolerance, and neutropenia. In addition, these drugs are limited in that they only help to regulate symptoms to an appropriate level rather than providing a fundamental treatment.

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

[0008] Cells release various membrane-type endoplasmic reticulum into the extracellular environment, and these released endoplasmic reticulum are usually called extracellular vesicles (EVs). Extracellular vesicles are also called cell membrane-derived endoplasmic reticulum, ectosomes, shedding vesicles, microparticles, exosomes, etc., and in some cases, they are used to distinguish them from exosomes.

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

[0010] In other words, exosomes, which are called avatars of cells, contain bioactive factors such as growth factors just like cells, but act as carriers that carry bioactive factors between cells, i.e., communicate 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 such as plants, bacteria, fungi, and algae. For example, exosomes can be isolated not only from culture fluids of cancer cells, immune cells, mesenchymal stem cells, etc., but also from culture fluids of plant cells or plant stem cells.

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

[0012] Catnip, whose scientific name is Nepeta cataria, is a perennial plant belonging to the Lamiales, Lamiaceae, and Nepeta genus. Nepeta cataria is usually called catnip because cats love it. Catnip is a type of herb, and when dried, it gives off a peppermint scent and is also called catnip or catmint. Catnip is native to Asia and Europe, and is distributed in Korea, Japan, China, Central Asia, Africa, Europe, North America, etc., and is known to have pain relieving and fever reducing effects. However, the technology so far is at the level of using extracts obtained by hot water extraction or solvent extraction of catnip leaves, stems, or the whole plant as food and cosmetic raw materials. However, solvent extracts have the problem of human toxicity due to residual extraction solvents.

[0013] The present inventors have confirmed that catnip-derived exosomes are effective in improving skin elasticity, reducing skin wrinkles, regenerating skin, acting as an anti-inflammator, and healing or promoting wound healing, and have developed a cosmetic composition for improving skin elasticity, reducing skin wrinkles or regenerating skin, and a pharmaceutical composition for anti-inflammatories, 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 the background art are merely intended to facilitate understanding of the background of the present invention, and are not cited as an admission that they may 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, improving skin wrinkles, regenerating skin, anti-inflammation, 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 illustrative and do not limit the scope of the present invention. In addition, 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, improving skin wrinkles, skin regeneration, anti-inflammation, wound healing or promoting wound healing, comprising exosomes derived from catnip (Nepeta cataria) as an active ingredient.

[0020] The term catnip used in this specification is a perennial plant belonging to the order Lamiales, family Lamiaceae, and genus Nepeta, and its scientific name is Nepeta cataria. Catnip is a type of herb, and when dried, it gives off a peppermint scent and is also called catnip or catmint. Catnip is native to Asia and Europe, and is distributed 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 secreted or released from plant cells into the extracellular space, and is also defined as exosome-like vesicles or exosome-like particles.

[0022] The term "skin elasticity" used herein means the property of skin that is easily restored to its original shape when deformed by an external force is removed. "Skin wrinkles" means fine wrinkles that occur when the skin is aged, 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" used herein means suppressing or inhibiting the formation of wrinkles in 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 due to bacterial infection, viral infection, or fungal infection, burns, inflammation due to burns, wounds, inflammation due to wounds, and the like.

[0024] As used herein, the term "wound" refers to a state in which an organism is injured, and includes pathological conditions in which tissues constituting the internal or external surface of an organism, such as skin, muscle, nerve tissue, bone, soft tissue, internal organs, or vascular tissue, are disrupted or destroyed. As non-limiting examples, wounds include abrasions, lacerations, punctures, cuts, avulsions, bed sores, bed sores, tissue destruction by radiation, penetrating wounds, gun shot wounds, burns, frostbite, surgical wounds, suture sites after plastic surgery, chemical wounds, and the like, and may include damage to any part of an individual.

[0025] In the present specification, the term "iontophoresis" refers to a method of passing a microcurrent through the skin to which an active substance is applied, creating a potential difference and changing the electrical environment of the skin, thereby allowing the ionized active ingredient to permeate the skin by electrical repulsion. The 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, a method in which a battery is attached to the electrode patch itself and a microcurrent is introduced into the skin, and a method in which a microcurrent is introduced into the skin through a patch equipped with a reversed electrodialysis means that generates a current due to the ion concentration difference between a high-concentration electrolyte solution and a low-concentration electrolyte solution. However, the present invention is not limited thereto, and it goes without saying that various types of iontophoresis can be used.

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

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

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

[0029] The composition containing the catnip (Nepeta cataria)-derived exosome of the present invention 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, improving skin wrinkles, or regenerating skin, comprising exosomes derived from catnip (Nepeta cataria) as an active ingredient. For example, the cosmetic composition may be 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 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 pharma- ceutically 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. In addition, the effective amount of the pharmaceutical composition of an embodiment of the present invention means the amount required for administration to expect 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, form, etc. of the additional component 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, based on the total amount of the injection. In addition, 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, administration time, and administration method. For example, when the pharmaceutical composition of one embodiment of the present invention is administered to an adult, it can be administered once or several times at a dose of 0.001 mg / kg to 100 mg / kg per day.

[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, UV absorbers, emulsifiers, surfactants, thickeners, alcohols, powder components, colorants, aqueous components, water, various skin nutrients, etc., can be appropriately blended as needed, within the scope that does not impair the effects of the present invention.

[0036] In addition, in the cosmetic composition according to one embodiment of the present invention, in addition to the exosomes derived from catnip (Nepeta cataria), a conventionally used skin improving agent and / or moisturizing agent may be mixed and used to the extent that the functions (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 an embodiment of the present invention can be used for the purpose of improving skin elasticity, improving skin wrinkles, regenerating skin, etc., and the cosmetic formulation can be prepared into any formulation commonly prepared in the art. For example, the cosmetic composition can be prepared into a patch, a mask pack, a sheet mask, a softening lotion, a nutritious lotion, an astringent lotion, a nutritious cream, a massage cream, an eye cream, a cleansing cream, an essence, an eye essence, a cleansing lotion, a cleansing foam, a cleansing water, a sunscreen, a lipstick, a soap, a shampoo, a surfactant-containing cleanser, a bath additive, a body lotion, a body cream, a body oil, a body essence, a body cleanser, a hair dye, a hair tonic, etc., but is not limited thereto.

[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 may be appropriately selected and blended by those skilled in the art without difficulty depending on the type or intended use of the cosmetic composition.

[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 the condition of the skin means improving / causing the condition of the skin or preventively regulating the condition of the skin, and improving the condition of the skin means a visually and / or tactilely perceptible positive change in the appearance and feel of the skin. For example, improving the condition of the skin may be improving wrinkles, improving elasticity of the skin, or regenerating the 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, a mask pack, or a sheet mask to which the cosmetic composition is applied or soaked, to the skin of a mammal, or sequentially carrying out (a) and (b). In the step (a), a lotion or cream can be used as the cosmetic composition.

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

[0043] As a non-limiting example, in the cosmetic method of one embodiment of the present invention, the cosmetic composition can 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 feline, human, canine, rodent, equine, bovine, simian, or porcine.

[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 area of ​​inflammation or a wound area.

[0046] As a non-limiting example, in the method for treating inflammation, treating wounds, 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 feline, human, canine, rodent, equine, bovine, simian, or porcine. Effect 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, improving skin wrinkles, regenerating skin, anti-inflammation, and wound healing or promoting wound healing.

[0049] On the other hand, the scope of the present invention is not limited by the above-mentioned effects. [Brief description 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 the catnip-derived exosomes of the present invention. [Diagram 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). [Diagram 3] 1 is a graph showing the relative collagen amount that increases in an exosome concentration-dependent manner 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. [Diagram 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] 1 is a graph showing that treatment of RAW264.7 cells with catnip-derived exosomes reduces LPS-induced IL-6. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[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 do not limit or restrict the scope of the present invention. Anything that can be easily inferred by a person skilled in the art to which the present invention belongs 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 the present invention are incorporated herein by reference.

[0052] Throughout the specification, when a part is described as "comprising" certain elements, this does not mean to exclude other elements, but rather that it may further include other elements, unless specifically stated to the contrary. EXAMPLES

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

[0054] Example 2: Purification of exosomes derived from catnip Catnip juice obtained in the same manner as in Example 1 was centrifuged and ultracentrifuged in stages to remove many impurities, and catnip-derived exosomes were isolated. The thawed catnip juice was centrifuged at 2,000×g for 10 minutes, and then filtered through a filter membrane containing 0.22 μm pores to obtain a filtrate. After that, it was subjected to a first centrifugation at 10,000×g and a second centrifugation at 20,000×g, and the supernatant was collected, and the supernatant was ultracentrifuged at 100,000×g for 70 minutes at 4° C. to obtain a pellet from the bottom. The obtained pellet was suspended in PBS (Phosphate Buffered Saline) to finally obtain catnip-derived exosomes. The size and concentration of catnip-derived exosomes isolated by the above method were confirmed by nanoparticle tracking analysis (NTA) using NS300 (purchased from Malvern Panalytical) (FIG. 1).

[0055] <Example 3: Confirmation of the transduction ability of catnip-derived exosomes to skin fibroblasts> In order to confirm whether catnip-derived exosomes are delivered into human skin fibroblasts (purchased from ATCC), the following analysis was performed. The membrane of the catnip-derived exosomes prepared in Example 2 was reacted with PKH67 (purchased from Sigma-Aldrich) fluorescent dye to fluorescently stain it. After the reaction, the reaction solution was fractionated with a MiniTrap-25 (purchased from Cytiva) column to remove free PKH67 fluorescent dye that was not stained in the exosome membrane. For the negative control group, PKH67 fluorescent dye was reacted with a buffer solution and then fractionated with a MiniTrap-25 column. The exosomes stained with PKH67 were cultured with human skin fibroblasts that had been previously cultured, and then it was observed using a fluorescent microscope whether the exosomes were delivered into the cells over time. Hoechst fluorescent dye (purchased from Thermo Fisher) was used to stain the cell nuclei, and CellMask Orange fluorescent dye (purchased from Thermo Fisher) was used to stain the cytoplasm. As a result of confirming whether exosomes were delivered into cells, it was confirmed that fluorescently stained exosomes were delivered into cells and green fluorescence accumulated inside the cells over time (Figure 2).

[0056] <Example 4: Confirmation of collagen production promotion effect> Human dermal fibroblasts (purchased from ATCC) dispersed in DMEM medium containing fetal bovine serum were dispensed into a multi-well plate, cultured for 24 hours, and then cultured in serum-free medium for another 48 hours. Then, the catnip-derived exosomes prepared in Example 2 were diluted in serum-free medium, and then applied to human dermal fibroblasts, which were then cultured. To confirm the efficacy of collagen production using human dermal fibroblasts, the experimental groups were classified 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 FIG. 3): an experimental group in which the 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 FIG. 3): an experimental group in which the catnip-derived exosomes prepared in Example 2 were diluted in serum-free medium and treated (treatment concentration: 4.0 × 10 10 particles / mL).

[0057] In each experimental group, human dermal fibroblasts were treated and cultured for 24 hours, and the culture medium was collected and centrifuged to prepare a 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 divided by the total cell number measured by an MTT assay kit (purchased from Sigma-Aldrich) to normalize and determine the relative collagen amount.

[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 ( FIG. 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 improving efficacy, wrinkle improvement efficacy, and / or skin regeneration efficacy.

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

[0061] <Example 5: Confirmation of skin regeneration efficacy using skin fibroblasts> To evaluate whether the catnip-derived exosomes prepared as in Example 2 promote wound healing in human dermal fibroblasts (Human Dermal Fibroblast; purchased from ATCC), a scratch-wound assay was performed. Human dermal fibroblasts dispersed in DMEM medium containing fetal bovine serum were dispensed into a wound induction culture plate (ImageLock Plate; purchased from EssenBio) at a density of 5,000 cells / well, cultured for 24 hours under conditions of 5% CO2 and 37°C to confirm a confluency of 90% or more, and then a scratch was induced using a WoundMaker (purchased from EssenBio). To confirm the efficacy of skin regeneration using human dermal fibroblasts, the experimental groups were classified as follows: (1) Negative control (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 the 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 the 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 of the experimental groups was treated with Scratch-Wound and human dermal fibroblasts were cultured at 5% CO2 and 37℃ for 12 hours, and the wound healing ability was measured using Incucyte (purchased from Sartorius).

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

[0064] From the above experimental results, it can be seen that the catnip-derived exosomes of the present invention have excellent efficacy in promoting the migration of human skin fibroblasts, i.e., excellent wound repair 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> In order to confirm whether catnip-derived exosomes are delivered into mouse macrophages (RAW264.7; purchased from ATCC), the following analysis was performed. The membrane of the catnip-derived exosomes prepared in Example 2 was reacted with PKH67 (purchased from Sigma Aldrich) fluorescent dye to fluorescently stain it. After the reaction, the reaction solution was fractionated with a MiniTrap-25 (purchased from Cytiva) column to remove free PKH67 fluorescent dye that was not stained in the exosome membrane. As a negative control group, PKH67 fluorescent dye was reacted with a buffer solution and then fractionated with a MiniTrap-25 column. The exosomes stained with PKH67 were cultured with mouse macrophages that had been previously cultured and prepared, and then it was observed using a fluorescent microscope whether the exosomes were delivered into the cells over time. Hoechst fluorescent dye (purchased from Thermo Fisher) was used to stain the cell nuclei, and CellMask Orange fluorescent dye (purchased from Thermo Fisher) was used to stain the cytoplasm. As a result of confirming whether exosomes were delivered into cells, it was confirmed that fluorescently stained exosomes were delivered into cells, and green fluorescence accumulated inside the cells over time (Figure 5).

[0067] Example 7: Evaluation of anti-inflammatory efficacy of catnip-derived exosomes In order to confirm whether the catnip-derived exosomes prepared as in Example 2 exhibit anti-inflammatory efficacy, the effect of the catnip-derived exosomes on the amount of IL-6 produced in mouse macrophages, RAW264.7 cells, was examined. RAW264.7 cells were suspended in DMEM medium containing 10% FBS, and the suspension was dispensed into each well of a multiwell plate to have a confluency of 80-90%. On the next day, catnip-derived exosomes diluted in a new medium containing LPS [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. The experimental groups for evaluating anti-inflammatory efficacy were classified as follows: (1) Negative control (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. The amount of IL-6 (inflammatory cytokine) produced in the group treated with LPS alone and the amount of IL-6 produced in the experimental groups treated with dexamethasone and catnip-derived exosomes (low and high concentrations) mixed with LPS medium were confirmed according to the ELSIA kit (purchased from R&D systems) manufacturer's manual (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 ( FIG. 6 ).

[0070] From the above experimental results, it was found 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 effectively used as an active ingredient of a pharmaceutical composition for anti-inflammatory, wound treatment and / or wound treatment promotion.

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

Claims

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

2. 2. The cosmetic composition for improving skin elasticity, improving skin wrinkles and regenerating skin according to claim 1, which is 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. The cosmetic composition for improving skin elasticity, improving skin wrinkles, and regenerating skin according to claim 1 or 2, wherein the catnip-derived exosomes are isolated from a catnip plant cell culture medium, a catnip callus culture medium, a catnip plant stem cell culture medium, a catnip juice, or an equivalent biological solution of catnip, or are derived from catnip plant cells or plant stem cells.

4. The cosmetic composition for improving skin elasticity, improving skin wrinkles, and regenerating skin according to claim 3, wherein the catnip-derived exosomes are separated and purified from catnip juice.

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

6. The pharmaceutical composition for anti-inflammation, wound healing or wound healing promotion according to claim 5, which is administered or administered by injection, microneedling, iontophoresis, application, or a combination thereof.

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

8. The pharmaceutical composition for anti-inflammatory, wound healing or wound healing promotion according to any one of claims 5 to 7, wherein the catnip-derived exosomes are isolated from catnip plant cell culture fluid, catnip callus culture fluid, catnip plant stem cell culture fluid, catnip juice, or equivalent biological solutions of catnip, or are derived from catnip plant cells or plant stem cells.

9. The pharmaceutical composition for anti-inflammation, wound healing or wound healing promotion according to claim 8 , wherein the catnip-derived exosomes are separated and purified from catnip juice.

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

11. The cosmetic method according to claim 10, 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 which has been soaked in the cosmetic composition, to the skin of a mammal; or sequentially carrying out (a) and (b).

12. The cosmetic method according to claim 11, wherein a lotion or cream is used as the cosmetic composition in step (a).

13. The cosmetic method according to claim 11 or 12, further comprising the step of: (c) removing the patch, mask pack or sheet mask from the skin of the mammal after the step (b), and applying the cosmetic composition to the skin of the mammal.

14. The cosmetic method according to claim 13, wherein in step (c), a lotion or cream is used as the cosmetic composition.

15. The cosmetic method according to any one of claims 10 to 12, wherein the mammal is a cat, a human, a dog, a rodent, a horse, a cow, a monkey, or a pig.

16. 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 an exosome derived from catnip (Nepeta cartaria) as an active ingredient to a mammal other than humans, or applying a pharmaceutical composition comprising an exosome derived from catnip (Nepeta cartaria) as an active ingredient to the skin, an inflammation site or a wound site of a mammal other than humans.

17. 17. The method for treating inflammation, treating wounds or promoting wound healing according to claim 16, wherein the pharmaceutical composition is administered by injection, microneedling, iontophoresis, or a combination thereof.

18. The method for treating inflammation, treating wounds or promoting wound healing according to claim 16, wherein the pharmaceutical composition is in the form of an injection, an infusion, a spray, a liquid or a patch.

19. The method for treating inflammation, treating wounds or promoting wound healing according to any one of claims 16 to 18, wherein the mammal is a cat, a dog, a rodent, a horse, a cow, a monkey or a pig.

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