Novel composition comprising actinidia polygama-derived exosomes as active ingredient

Abstract

The present invention provides a composition for skin elasticity improvement, skin wrinkle reduction, skin regeneration, skin whitening, anti-inflammation, wound healing or wound healing promotion, comprising exosomes derived from Actinidia polygama as an active ingredient. Compared to a conventional Actinidia polygama hot water extract, Actinidia polygama solvent extract and filtrate of the extracts, the composition of the present invention is less likely to contain residual solvents, other impurities and the like, and has the excellent effects of skin elasticity improvement, skin wrinkle reduction, skin regeneration, skin whitening, anti-inflammation, wound healing, or wound healing promotion.

Description

A novel composition containing exosomes derived from ginseng as an active ingredient

[0001] The present invention relates to a novel composition comprising exosomes derived from Actinidia polygama as an active ingredient, and more specifically, to a composition for improving skin elasticity, improving skin wrinkles, regenerating skin, healing wounds, or promoting wound healing, comprising exosomes derived from Actinidia polygama as an active ingredient.

[0002] In addition, the present invention relates to a cosmetic composition for improving skin elasticity, improving skin wrinkles, or regenerating skin, and a pharmaceutical composition for healing or promoting wound healing, comprising the composition.

[0003] As skin aging occurs, skin elasticity decreases and skin wrinkles increase. It is known that the decrease in skin elasticity and the formation of skin wrinkles 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] In addition, it is known that the synthesis of prostaglandin E2 increases due to an increase in COX-2, an enzyme that produces inflammatory cytokines in skin cells, as a result of the progression of aging or ultraviolet rays, and the production of inflammatory factors increases. Inflammatory reactions increase the biosynthesis of MMPs, which causes collagen breakdown, resulting in decreased skin elasticity and the formation of wrinkles. In particular, when sunlight and ultraviolet rays are directly irradiated on the skin, a large amount of free radicals are generated, and these free radicals damage the skin's antioxidant defense system, accelerating skin aging by increasing wrinkles and loosening the skin. Substances known to be effective in improving skin wrinkles include adenosine and retinoic acid, but adenosine has minimal clinical efficacy, and retinoic acid cannot be used by women of childbearing age and has side effects such as erythema.

[0005] Meanwhile, recent studies have reported that cell secretomes contain various bioactive factors that regulate cell behavior, and in particular, cell secretomes contain 'exosomes' that have intercellular signaling functions, and research on their components and functions is actively underway.

[0006] Cells release various membrane-bound vesicles into the extracellular environment, commonly referred to as extracellular vesicles (EVs). EVs are also known as membrane-derived vesicles, ectosomes, shedding vesicles, microparticles, and exosomes, and are sometimes used interchangeably with exosomes.

[0007] Exosomes are vesicles measuring tens to hundreds of nanometers in size with a double phospholipid membrane that has the same structure as the cell membrane, and contain proteins and nucleic acids (mRNA, miRNA, etc.), called exosome cargo, inside. Exosome cargo includes a wide range of signaling factors, and these signaling factors are known to be cell type-specific and regulated differently depending on the environment of the secreting cell. Exosomes are intercellular signaling mediators secreted by cells, and various cell signals transmitted through them are known to regulate cell 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 cell from which they are derived. In the case of proliferating stem cell-derived exosomes, they regulate cell behaviors such as cell migration, proliferation, and differentiation, and reflect the characteristics of stem cells related to tissue regeneration (Nature Review Immunology 2002 (2) 569-579).

[0008] That is, exosomes, which are called avatars of cells, contain bioactive factors such as growth factors similar to cells, and act as carriers that transport bioactive factors between cells, that is, they act as 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 such as plants, bacteria, fungi, and algae. For example, exosomes can be isolated from cultures of cancer cells, immune cells, mesenchymal stem cells, etc., as well as from cultures of plant cells or plant stem cells.

[0009] However, research on the isolation, purification, and characterization of exosomes derived from plant cells or plant stem cells is still in its infancy. Most of these efforts simply market exosomes as extracellular vesicles mixed within plant juice filtrates. Therefore, more in-depth characterization and functional studies on exosomes derived from plant cells are needed.

[0010] Actinidia polygama is a deciduous climbing plant belonging to the Actinidia genus of the Actinidiaceae family. Native to Korea, Siberia, and Japan, Actinidia polygama is known to be effective for gout, arthritis, lumbago, chills, and colds. However, current technology only utilizes extracts obtained through hot water or solvent extraction of the leaves, stems, or fruits of Actinidia polygama, which are used as food or cosmetic ingredients. However, solvent extraction poses the problem of human health hazards due to residual extraction solvent.

[0011] The present inventors have confirmed that exosomes derived from kiwifruit are effective in improving skin elasticity, improving skin wrinkles, skin regeneration, wound healing, or promoting wound healing, and have developed a cosmetic composition for improving skin elasticity, improving skin wrinkles, or skin regeneration, and a pharmaceutical composition for wound healing or promoting wound healing, which contain exosomes derived from kiwifruit as an effective ingredient.

[0012] Meanwhile, it should be understood that the matters described as background art above are only intended to enhance understanding of the background of the present invention, and are not cited as an admission that they can be used as “prior art” of the present invention.

[0013] The purpose of the present invention is to provide a composition for improving skin elasticity, improving skin wrinkles, regenerating skin, healing wounds, or promoting wound healing, which contains exosomes derived from Actinidia polygama as an active ingredient.

[0014] In addition, the purpose of the present invention is to provide a cosmetic composition for improving skin elasticity, improving skin wrinkles, or regenerating skin, and a pharmaceutical composition for healing or promoting wound healing, including the composition.

[0015] However, the aforementioned tasks of the present invention are exemplary and do not limit the scope of the present invention. Furthermore, other objectives and advantages of the present invention will become more apparent through the detailed description, claims, and drawings below.

[0016] The present invention provides a composition for improving skin elasticity, improving skin wrinkles, regenerating skin, whitening skin, anti-inflammation, wound healing or promoting wound healing, which contains exosomes derived from Actinidia polygama as an active ingredient.

[0017] As used herein, the term "Actinidia polygama" is a deciduous climbing plant belonging to the Actinidiaceae family and the genus Actinidia, and does not refer only to the fruit of the Actinidia polygama tree, but is used to encompass the Actinidia tree, its leaves, stems, roots, bark, flowers, and fruits. Meanwhile, the fruit of the Actinidia polygama that has a lump-like growth due to insects attached to it is called a "fruit gall." As used herein, the term "Actinidia polygama" includes galls.

[0018] 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 are also defined as exosome-like vesicles or exosome-like particles.

[0019] The term "skin elasticity" as used herein refers to the property of skin that has been deformed by an external force, easily returning to its original shape when the external force is removed. "Skin wrinkles" refer to fine lines formed when the skin weakens, and can be caused by genetic factors, a decrease in collagen and elastin in the skin's dermis, or external factors. 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.

[0020] The term "wound" as used herein refers to a state in which a living body is damaged, and encompasses a pathological state in which tissues forming the internal or external surface of a living body, such as skin, muscle, nerve tissue, bone, soft tissue, internal organ, or vascular tissue, are divided or destroyed. As non-limiting examples of the present invention, wounds include abrasions, lacerations, puncture wounds, cuts, avulsions, bedsores, ulcers, destruction of tissue by radiation, penetrated wounds, gunshot wounds, burns, frostbite, surgical wounds, suture sites after plastic surgery, chemical wounds, etc., and may include damage to any part of an object.

[0021] The term "iontophoresis" used herein refers to a method of changing the electrical environment of the skin by applying a potential difference to a microcurrent flowing through the skin to which an active ingredient has been applied, thereby allowing the ionized active ingredient to penetrate the skin through electrical repulsion. The iontophoresis used in one specific example 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 to introduce a microcurrent into the skin, a method in which a battery is installed in the electrode patch itself to introduce a microcurrent into the skin, a method in which a microcurrent is introduced into the skin through a patch equipped with a reversed electrodialysis means that generates a current through the difference in ion concentration between a high-concentration electrolyte solution and a low-concentration electrolyte solution, etc. However, the present invention is not limited thereto, and it goes without saying that various methods of iontophoresis may be used.

[0022] The term "exosome derived from Actinidia polygama" as used in the specification of the present invention is meant to include all exosomes isolated from, for example, Actinidia polygama plant cell culture, Actinidia callus culture, Actinidia plant stem cell culture, Actinidia extract, or biological solutions of Actinidia equivalent thereto, or derived from, for example, secreted and / or released Actinidia polygama plant cells or plant stem cells.

[0023] In a composition of one specific example of the present invention, the exosomes derived from the kiwifruit may be isolated and purified from a callus culture derived from the leaves, stems, roots, flowers, petals, fruits and / or galls of the kiwifruit, or may be isolated and purified from the juice of the leaves, stems, whole plants, bark, fruits and / or galls.

[0024] A composition comprising an exosome derived from Actinidia polygama as an active ingredient according to one specific example of the present invention may exhibit at least one of the effects of improving skin elasticity, improving skin wrinkles, regenerating skin, whitening skin, anti-inflammation, wound healing, or promoting wound healing.

[0025] The composition comprising the exosome derived from Actinidia polygama of the present invention as an active ingredient may be a cosmetic composition or a pharmaceutical composition.

[0026] The present invention provides a cosmetic composition for improving skin elasticity, improving skin wrinkles, whitening skin, or regenerating skin, comprising exosomes derived from Actinidia polygama 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.

[0027] In addition, the present invention provides a pharmaceutical composition for anti-inflammation, wound healing or wound healing promotion, which comprises an exosome derived from Actinidia polygama as an active ingredient.

[0028] As a non-limiting example of the present invention, the pharmaceutical composition of one embodiment of the present invention may be administered or treated by injection, microneedling, iontophoresis, application, or a combination thereof. For example, the pharmaceutical composition may be in the form of an injection, an infusion, a spray, a liquid, or a patch.

[0029] When the composition of one embodiment of the present invention is used as a pharmaceutical composition, it may include a pharmaceutically acceptable carrier, excipient, or diluent, etc. 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, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil. In addition, the effective amount of the pharmaceutical composition of one embodiment of the present invention means the amount required for administration to expect an anti-inflammatory, wound healing, or wound healing promoting effect.

[0030] The mixing 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 components as described above. For example, the pharmaceutical composition of the present invention can be included in an amount of about 0.1 to 99 wt%, preferably about 10 to 90 wt%, of 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 formulation, the formulation method, the patient's age, sex, weight, health condition, diet, excretion rate, administration time, and administration method. For example, when administering the pharmaceutical composition of one embodiment of the present invention 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.

[0031] Meanwhile, when the composition of one specific example of the present invention is manufactured into a cosmetic composition, components commonly used in cosmetic compositions, such as moisturizers, antioxidants, oily components, ultraviolet absorbers, emulsifiers, surfactants, thickeners, alcohols, powder components, coloring agents, aqueous components, water, various skin nutrients, etc., can be appropriately mixed as needed within a range that does not impair the effects of the present invention.

[0032] In addition, the cosmetic composition of one specific example of the present invention may be used by mixing together with a conventionally used skin improvement agent and / or moisturizer, in addition to the exosome derived from the dogwood, as long as the effect (e.g., improvement of skin condition, improvement of skin elasticity, improvement of skin wrinkles, skin regeneration, etc.) is not impaired.

[0033] The cosmetic composition of one specific example of the present invention can be applied in various forms, such as a patch, a mask pack, a mask sheet, 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, and a blotting paper.

[0034] The cosmetic composition of one specific example of the present invention can be used for the purpose of improving skin elasticity, improving skin wrinkles, whitening skin, and regenerating skin, and the cosmetic formulation can be manufactured into any formulation commonly manufactured in the art. For example, the cosmetic composition can be formulated into a patch, a mask pack, a mask sheet, an emollient toner, a nourishing toner, an astringent toner, a nourishing 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 bathing agent, a body lotion, a body cream, a body oil, a body essence, a body cleanser, a hair dye, a hair tonic, and the like, but is not limited thereto.

[0035] A cosmetic composition according to one embodiment of the present invention comprises ingredients commonly used in cosmetic compositions, and may include conventional adjuvants such as antioxidants, stabilizers, solubilizers, vitamins, pigments, and fragrances, as well as carriers. Furthermore, for each formulation of the cosmetic composition, other ingredients can be appropriately selected and blended without difficulty by those skilled in the art, depending on the type or intended use of the cosmetic composition.

[0036] Another embodiment of the present invention provides a cosmetic method for regulating the skin condition of a mammal, excluding therapeutic use, using the cosmetic composition. In the cosmetic method of the present invention, "regulating skin condition" refers to improving and / or preventatively regulating skin condition, and "improving skin condition" refers to a positive change in the appearance and feel of the skin that can be visually and / or tactilely perceived. For example, "improving skin condition" may include improving wrinkles, improving skin elasticity, whitening, or regeneration.

[0037] A cosmetic method of one specific example of the present invention comprises (a) directly applying the cosmetic composition to the skin of a mammal, (b) contacting or attaching a patch, mask pack, or mask sheet on which the cosmetic composition has been applied or deposited, to the skin of a mammal, or sequentially performing (a) and (b). In step (a), a lotion or cream may be used as the cosmetic composition.

[0038] In addition, the cosmetic method of one specific example of the present invention may further include, after step (b), a step (c) of removing the patch, mask pack, or mask sheet from the skin of the mammal and applying the cosmetic composition to the skin of the mammal. In step (c), a lotion or cream may be used as the cosmetic composition.

[0039] As an example and not limiting example of the present invention, in one specific cosmetic method 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 formulation, a liquid formulation, or a patch formulation.

[0040] In one specific example of the cosmetic method of the present invention, the mammal may be a human, a dog, a cat, a rodent, a horse, a cow, a monkey, or a pig.

[0041] The present invention also provides a method for anti-inflammation, wound healing or accelerating wound healing, comprising administering a therapeutically effective amount of the pharmaceutical composition to a mammal or applying the pharmaceutical composition to the skin or a wound site.

[0042] As a non-limiting example of the present invention, in a method for anti-inflammation, wound healing, or wound healing promotion 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, an infusion, a spray, a liquid, or a patch.

[0043] In one embodiment of the method for anti-inflammation, wound treatment or accelerating wound healing of the present invention, the mammal may be a human, a dog, a cat, a rodent, a horse, a cow, a monkey, or a pig.

[0044] The composition of the present invention has a low possibility of containing residual solvents, other impurities, etc. compared to conventional water extracts of perilla, solvent extracts of perilla, and filtrates of these extracts, and has excellent effects of improving skin elasticity, improving skin wrinkles, regenerating skin, healing wounds, or promoting wound healing.

[0045] Meanwhile, the scope of the present invention is not limited by the effects described above.

[0046] Figure 1 is a graph showing the particle size distribution and particle number obtained by performing NTA (nanoparticle tracking analysis) on exosomes derived from the dogwood of the present invention.

[0047] Figure 2 is a cell fluorescence microscope image confirming that fluorescently dyed exosomes derived from dandelion are delivered to human skin fibroblasts (green: exosomes delivered into cells, blue: cell nuclei).

[0048] Figure 3 is a graph showing the relative amount of collagen that increases in a concentration-dependent manner when human skin fibroblasts are treated with exosomes derived from kiwifruit.

[0049] Figure 4 is a graph showing that the migration of human skin fibroblasts increases after treating scratch-wounds with exosomes derived from ginseng.

[0050] The present invention is described in more detail in the following examples. However, the following examples are intended to illustrate the content of the present invention and do not limit or restrict the scope of the invention. Anything readily inferred by those skilled in the art from the detailed description and examples of the present invention is construed as falling within the scope of the present invention. References cited herein are incorporated herein by reference.

[0051] Throughout the specification, whenever a part is said to "include" a component, this does not mean that it excludes other components, but rather that it may include other components, unless otherwise stated.

[0052]

[0053] Example

[0054] Example 1: Preparation of persimmon juice

[0055] The leaves and / or stems of Actinidia polygama were first washed with running water, then with 70% ethanol, and then washed again with running water to remove any contaminants attached to the leaves and / or stems. The leaves and / or stems of Actinidia polygama were mixed with DBPS (Dulbecco's Phosphate Buffered Saline) and then juiced with a low-speed screw to obtain liquid juice, which was then filtered through a mesh screen to remove large floating substances.

[0056]

[0057] Example 2: Purification of exosomes derived from kiwifruit

[0058] The extract of the persimmon obtained in the same manner as in Example 1 was subjected to the following step-by-step centrifugation and ultracentrifugation methods to remove numerous impurities and to isolate exosomes derived from the persimmon.

[0059] The extract of the wild chives was centrifuged at 2,000×g for 10 minutes and then filtered through a membrane containing 0.22 μm pores to obtain a filtrate. Thereafter, the supernatant was recovered after a primary centrifugation at 10,000×g for 10 minutes at 25°C and a secondary centrifugation at 20,000×g for 30 minutes at 4°C. The supernatant filtered through a membrane containing 0.22 μm pores was subjected to ultracentrifugation at 100,000×g for 70 minutes at 4°C to obtain a pellet from the bottom of the centrifuge tube. The obtained pellet was suspended in PBS (Phosphate Buffered Saline) to finally obtain wild chive-derived exosomes. The size and concentration of exosomes derived from the perilla separated using the above method were confirmed by nanoparticle tracking analysis (NTA) using NS300 (purchased from Malvern Panalytical) (Fig. 1).

[0060]

[0061] Example 3: Confirmation of the transduction ability of exosomes derived from kiwifruit to skin fibroblasts.

[0062] To determine whether exosomes derived from kiwifruit were delivered into human dermal fibroblasts (purchased from ATCC), the following assay was performed. The membrane of the kiwifruit-derived exosomes prepared in Example 2 was reacted with the fluorescent dye PKH67 (purchased from Sigma-Aldrich) to fluorescently stain it. After the reaction, the reaction solution was fractionated using a MiniTrap-25 (purchased from Cytiva) column to remove free PHK67 (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 exosomes stained with PKH67 were cultured with pre-cultured human dermal fibroblasts, and the intracellular delivery of the exosomes over time was observed using a fluorescence microscope. Hoechst fluorescent dye (purchased from Thermo Fisher) was used to stain the nucleus of cells, and CellMask Orange fluorescent dye (purchased from Thermo Fisher) was used to stain the cytoplasm. As a result of examining whether exosomes were delivered into cells, it was confirmed that fluorescently dyed exosomes were delivered into cells, and green fluorescence accumulated inside the cells over time (Fig. 2).

[0063]

[0064] Example 4: Confirmation of collagen production promotion effect

[0065] Human dermal fibroblasts (purchased from ATCC) dispersed in DMEM medium containing fetal bovine serum were seeded in a multiwell plate at a density of 5 × 10 4 After seeding at a density of 10 cells / well, cells were cultured for 24 hours and then cultured overnight in serum-free medium. To confirm the efficacy of collagen production using human skin fibroblasts, the experimental groups were divided as follows:

[0066] (1) Negative control group (indicated as “NC” in Fig. 3): an experimental group in which only serum-free medium was treated to human skin fibroblasts;

[0067] (2) Positive control group (indicated as “PC” in Figure 3): An experimental group in which human skin fibroblasts were treated with culture medium containing 10% fetal bovine serum;

[0068] (3) High-concentration exosome treatment group derived from kiwifruit (indicated as “high-concentration exosome” in Fig. 3): Experimental group treated by diluting the kiwifruit-derived exosomes prepared in Example 2 in serum-free medium (treatment concentration: 3.0×10 9 particles / mL);

[0069] (4) Medium concentration exosome treatment group derived from kiwifruit (indicated as “medium concentration exosome” in Figure 3): Experimental group treated by diluting the kiwifruit-derived exosomes prepared in Example 2 in serum-free medium (treatment concentration: 9.0×10 8 particles / mL); and

[0070] (5) Low-concentration exosome treatment group derived from kiwifruit (indicated as “low-concentration exosome” in Figure 3); Experimental group in which the kiwifruit-derived exosomes prepared in Example 2 were diluted in serum-free medium and treated on human skin fibroblasts (treatment concentration: 3.0×10 8 particles / mL).

[0071] According to each of the above experimental groups, the corresponding substances were treated to human skin fibroblasts, cultured for 24 hours, and the culture medium was collected and centrifuged, and the centrifuged culture medium was prepared. The amount of collagen synthesized from human skin fibroblasts and accumulated in the culture medium was measured using an EIA kit for procollagen type I C-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 amount.

[0072] As a result, it was confirmed that exosomes derived from ginseng significantly increased collagen synthesis in human skin fibroblasts in a concentration-dependent manner compared to the negative control group (Fig. 3).

[0073] From the above experimental results, it can be seen that exosomes derived from perilla have excellent effects in increasing collagen synthesis, i.e., improving skin elasticity, improving wrinkles, and / or regenerating skin.

[0074] According to the above results, it can be seen that exosomes derived from kiwifruit have a useful functional activity, namely, the activity of increasing collagen synthesis, as a cosmetic product for improving skin elasticity, improving wrinkles, and / or regenerating skin. Therefore, exosomes derived from kiwifruit can be usefully utilized as an effective ingredient in cosmetic compositions for improving skin elasticity, improving wrinkles, and / or regenerating skin.

[0075]

[0076] Example 5: Confirmation of skin regeneration efficacy using skin fibroblasts.

[0077] To evaluate whether the exosomes derived from the quince prepared as in Example 2 promote wound healing in human dermal fibroblasts (purchased from ATCC), a scratch-wound assay was performed. Human dermal fibroblasts dispersed in DMEM medium containing fetal bovine serum were seeded in a wound-inducing culture plate (ImageLock Plate; purchased from EssenBio) at a density of 4 × 10 4 Cells were seeded at a density of 10 cells / well and cultured for 24 hours under 5% CO2 and 37°C. After confirming a confluence of 90% or more, scratches were induced using a WoundMaker (purchased from EssenBio). To confirm the skin regeneration efficacy using human skin fibroblasts, the experimental groups were divided as follows:

[0078] (1) Negative control group (indicated as “NC” in Figure 4): experimental group treated only with serum-free medium;

[0079] (2) Vehicle (indicated as “VC” in Figure 4): Experimental group treated by mixing DBPS (Dulbecco's Phosphate Buffered Saline) into serum-free medium;

[0080] (3) Positive control group (indicated as “PC” in Fig. 4): Experimental group treated with culture medium containing 10% fetal bovine serum;

[0081] (4) Group treated with extract of persimmon juice (indicated as “extract” in Fig. 4): The extract of persimmon juice prepared in Example 1 was diluted in serum-free medium and treated as an experimental group (treatment concentration: 2×10 9 particles / mL); and

[0082] (5) Exosome-treated group derived from kiwifruit (indicated as “exosome” in Figure 4); Experimental group treated by diluting exosomes derived from kiwifruit prepared in Example 2 in serum-free medium (treatment concentration: 3.0×10 9 particles / mL).

[0083] Afterwards, the corresponding substance was treated on the scratch-wound according to each experimental group above, and human skin fibroblasts were cultured for 24 hours under 5% CO2, 37℃ conditions, and the wound healing ability was measured using Incucyte (purchased from Sartorius).

[0084] As a result of measuring the wound healing ability, it was confirmed that the exosomes derived from the kiwifruit increased the migration of human skin fibroblasts compared to the negative control group and vehicle (Fig. 4). In addition, the wound area in the kiwifruit-derived exosome-treated group was reduced by approximately 59% compared to the vehicle, and the wound area in the kiwifruit juice-treated group was reduced by approximately 47% compared to the vehicle. Therefore, the exosomes derived from the kiwifruit are superior to the kiwifruit juice in wound healing ability and skin regeneration efficacy.

[0085] From the above experimental results, it can be seen that exosomes derived from perilla have excellent effects on promoting the migration of human skin fibroblasts, i.e., wound healing and skin regeneration effects.

[0086] Therefore, exosomes derived from perilla can be usefully utilized as an effective ingredient in a cosmetic composition for improving skin elasticity, improving wrinkles, and / or regenerating skin, and a pharmaceutical composition for treating or promoting wound healing.

[0087] While the present invention has been described with reference to the above-described embodiments, the present invention is not limited thereto. Those skilled in the art will appreciate that modifications and variations can be made without departing from the spirit and scope of the present invention, and that such modifications and variations also fall within the scope of the present invention.

Claims

1. A cosmetic composition comprising exosomes derived from Actinidia polygama as an active ingredient.

2. In paragraph 1, A cosmetic composition that exhibits at least one of the effects of improving skin elasticity, improving wrinkles, or regenerating skin.

3. In paragraph 1 or 2, A cosmetic composition comprising a shampoo, soap, rinse, surfactant-containing cleansing, cream, lotion, ointment, tonic, treatment, conditioner, suspension, emulsion, paste, gel, oil, wax, spray, aerosol, mist, or powder.

4. A pharmaceutical composition for wound healing or promoting wound healing, comprising exosomes derived from Actinidia polygama as an active ingredient.

5. In paragraph 4, A pharmaceutical composition for wound healing or promoting wound healing, administered or treated by injection, microneedling, iontophoresis, application, or a combination thereof.

6. In paragraph 4, A pharmaceutical composition for wound healing or promoting wound healing, in the form of an injection, infusion, spray, liquid or patch.

7. A cosmetic method for improving skin elasticity, improving skin wrinkles, or regenerating skin in a mammal, excluding therapeutic purposes, using a cosmetic composition containing exosomes derived from Actinidia polygama as an active ingredient.

8. In paragraph 7, A cosmetic method comprising: (a) directly applying the cosmetic composition to the skin of a mammal, (b) contacting or attaching a patch, mask pack or mask sheet on which the cosmetic composition has been applied or deposited, to the skin of a mammal, or performing (a) and (b) sequentially.

9. In paragraph 8, A cosmetic method in which a lotion or cream is used as a cosmetic composition in the step (a).

10. In clause 8 or 9, (c) A cosmetic method further comprising the step of removing the patch, mask pack or mask sheet from the skin of the mammal after step (b) and applying the cosmetic composition to the skin of the mammal.

11. In paragraph 10, A cosmetic method in which a lotion or cream is used as a cosmetic composition in the step (c) above.

12. In any one of paragraphs 7 to 9, The above mammal is a human, dog, cat, rodent, horse, cow, monkey, or pig, a cosmetic method.

13. A method for treating a wound or promoting wound healing, comprising a step of administering a therapeutically effective amount of a pharmaceutical composition comprising an exosome derived from Actinidia polygama as an active ingredient to a mammal other than a human, or a step of applying a pharmaceutical composition comprising an exosome derived from Actinidia polygama as an active ingredient to the skin or a wound site.

14. In paragraph 13, A method for wound healing or promoting wound healing, wherein the pharmaceutical composition is administered by injection, microneedling, iontophoresis or a combination thereof.

15. In paragraph 13, A method for treating a wound or promoting wound healing, wherein the pharmaceutical composition is in the form of an injection, infusion, spray, liquid or patch.

16. In any one of paragraphs 13 to 15, A method for treating a wound or promoting wound healing, wherein the mammal is a dog, a cat, a rodent, a horse, a cow, a monkey, or a pig.

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