Anti-aging and antioxidant cosmetic composition containing broccoli exosomes as an active ingredient, and functional cosmetics containing the same

Abstract

The present invention relates to a functional cosmetic composition containing broccoli exosomes as an active ingredient and functional cosmetics containing the same. Broccoli exosomes have a lipid bilayer structure and are absorbed into skin cells. The absorbed exosomes remove reactive oxygen species within skin cells, stimulate various antioxidant signaling mechanisms, exert antioxidant effects, and promote collagen and hyaluronic acid synthesis. Therefore, they are useful for producing functional cosmetics for skin anti-aging, skin whitening, wrinkle reduction, and skin cell regeneration.

Description

Detailed Description of the Invention

[0001] [Technical field] This application claims priority to Korean Patent Application No. 10-2020-0099518, filed on August 7, 2020, the entire contents of which are incorporated herein by reference in their entirety in the specification and drawings.

[0002] The present invention relates to an anti-aging, anti-oxidant and anti-inflammatory cosmetic composition, and a functional cosmetic comprising the same.

[0003] [Background technology] There are various causes of skin aging, but the most important cause is reactive oxygen species generated through ultraviolet rays and breathing. Reactive oxygen species are generated during normal metabolic processes, but are produced in excess when the body is in a diseased state or under high physical stress. Reactive oxygen species are also generated by exposure to ultraviolet rays. Reactive oxygen species lead to damage to skin cells and tissues. Normally, they kill abnormal and aged cells to maintain balance, but when they are generated in excess, they destroy the skin's antioxidant defense network, causing lipid peroxidation, protein oxidation, and destruction of intercellular components, as well as destruction of skin proteins such as collagen and elastin, which results in skin aging such as loss of elasticity, wrinkles, and pigmentation.

[0004] Skin proteins, including the connective tissue in the dermis, play an important role in maintaining skin elasticity and moisture, which is believed to be due to proteins that are essential for cell formation and production. Recent research has demonstrated a positive correlation between protein intake and increased skin moisture, and it can be inferred that nutritionally, the intake of high-quality protein is closely related to skin health. However, the effect of protein intake through food on the skin is only slight.

[0005] More than 500 types of plant extracts with specific pharmacological effects such as antioxidant and anti-aging are known in Korea, but their effects on the skin have hardly been verified, and since many ingredients exist in plant extracts at various ratios, it is difficult to find an effective substance with a certain effect. In addition, in plant ingredients, an effective active substance may act alone as a single ingredient, but may also exert its effect through the interaction of multiple ingredients. Furthermore, it is known that conventional various plant extracts have difficulty passing through the skin barrier, and the effect is limited when they are simply applied to the skin.

[0006] A number of documents are referenced and cited throughout this specification, the disclosures of which are incorporated herein by reference in their entirety in order to more clearly describe the state of the art to which the present invention pertains and the contents of the present invention.

[0007] Summary of the Invention [Problem to be solved by the invention] The present invention relates to the anti-aging, antioxidant, and anti-inflammatory effects of exosomes isolated and purified from broccoli, and aims to provide a cosmetic composition containing broccoli exosomes, which have anti-aging, antioxidant, and anti-inflammatory functions, as an active ingredient.

[0008] Another object of the present invention is to provide a functional cosmetic product for skin protection, wrinkle improvement, skin whitening, etc., which is produced by using the composition as a cosmetic raw material.

[0009] Another object of the present invention is to provide a method for antioxidant treatment of skin, comprising applying a cosmetic composition containing broccoli exosomes to a subject.

[0010] Another object of the present invention is to provide a cosmetic use of a cosmetic composition containing broccoli exosomes for skin antioxidant purposes.

[0011] Another object of the present invention is to provide a cosmetic use of a cosmetic composition containing broccoli exosomes for skin whitening, wrinkle improvement, and skin cell regeneration.

[0012] More specifically, the present invention aims to provide the following embodiments.

[0013] Example 1. A composition or cosmetic composition comprising broccoli exosomes; a method for anti-aging, anti-oxidation, skin whitening, anti-wrinkle, skin cell regeneration, moisturizing, improving skin barrier or anti-inflammation of skin, comprising applying a composition comprising broccoli exosomes or isolated broccoli exosomes to a subject; use or cosmetic use of a composition comprising broccoli exosomes or isolated broccoli exosomes for anti-aging, anti-oxidation, skin whitening, anti-wrinkle, skin cell regeneration, moisturizing, improving skin barrier or anti-inflammation of skin; or use of a composition comprising broccoli exosomes or isolated broccoli exosomes for the manufacture of a cosmetic for anti-aging, anti-oxidation, skin whitening, anti-wrinkle, skin cell regeneration, moisturizing, improving skin barrier or anti-inflammation of skin.

[0014] Embodiment 2. The composition; method; use; or application of embodiment 1, wherein the broccoli exosomes are derived from one or more selected from the group consisting of broccoli sprouts, broccoli flowers, and broccoli stems.

[0015] In any one of the preceding embodiments, the broccoli exosomes are present in an amount of 10 per mL of the composition. 6 ~10 pieces 13 A composition; method; use; or application comprising the content.

[0016] Embodiment 4. In any one of the preceding embodiments, the composition is in a dosage form selected from the group consisting of an aqueous solution, a suspension, an emulsion, a cream, a frozen solution, a spray-dried powder, and a freeze-dried powder.

[0017] Embodiment 5. Any one of the preceding embodiments, wherein the broccoli exosomes comprise polyphenols. A composition; method; use; or application.

[0018] Embodiment 6. In any one of the preceding embodiments, the composition has anti-aging or anti-oxidation uses; a composition; a method; a use; or an application.

[0019] Embodiment 7. A cosmetic product comprising a composition according to any one of the preceding embodiments.

[0020] Embodiment 8. The cosmetic product of any one of the preceding embodiments, wherein the cosmetic product is in a dosage form selected from the group consisting of mist, serum, nutrient lotion, softener, softener, emulsion, skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisture lotion, nourishing lotion, massage cream, nourishing cream, moisture cream, hand cream, foundation, powder, makeup base, essence, nourishing essence, pack, soap, cleansing foam, cleansing lotion, cleansing cream, body lotion, body cleanser, facial cleanser, treatment, beauty serum, beauty pack, ointment, gel, liniment, liquid, patch, spray, bath additive, sunscreen, sun oil, and hair product.

[0021] Embodiment 9. In any one of the preceding embodiments, the cosmetic is a functional cosmetic having one or more effects selected from the group consisting of anti-skin aging, skin whitening, wrinkle improvement, skin cell regeneration, moisturizing, skin barrier improvement, and anti-inflammation.

[0022] Embodiment 10. A method for producing a composition or cosmetic composition according to any one of the previous embodiments, comprising the steps of crushing broccoli; centrifuging; and performing tangential flow filtration (TFF).

[0023] Other objects and advantages of the present invention will become more apparent from the following detailed description of the invention, the claims and the drawings.

[0024] [Means for solving the problems] One aspect of the present invention provides a cosmetic composition containing broccoli sprout exosomes as an active ingredient.

[0025] The term "broccoli exosome" as used in the present invention generally refers to exosomes extracted from broccoli having a size of about 50 to 300 nm, preferably 50 to 200 nm, or extracellular vesicles containing exosomes. Broccoli exosomes form particles of uniform size of 50 to 200 nm made of lipid bilayer membrane.

[0026] Although various plant extracts have been known to contain anti-aging and antioxidant substances, there are problems with their application as cosmetic raw materials, as they are difficult to pass through the skin barrier in the form of water-soluble extracts, and the active ingredients are easily exposed to the extraction solvent and destroyed during the extraction process. On the other hand, exosomes are nanoparticles made of lipid bilayer membranes that are absorbed into skin cells through receptor-mediated endocytosis and membrane fusion, and the active ingredients contained within are not directly exposed to the outside, so they can maintain their biological activity for a long period of time without being altered, making them very advantageous for commercial use in the cosmetics industry.

[0027] In the present invention, broccoli exosomes have the function of suppressing active oxygen that causes skin aging, pigmentation, and inflammation due to the anti-aging and antioxidant components derived from broccoli, which is the raw material including broccoli sprouts, and therefore serve the purpose of anti-aging and / or antioxidant. Specifically, broccoli exosomes containing polyphenols and antioxidant enzyme promoters suppress active oxygen accumulated in skin cells, and show efficacy in skin regeneration, wrinkle improvement, whitening, and inflammation relief.

[0028] Broccoli, which is a raw material plant of the present invention, includes Brassica oleracea L. The raw material plant part may be the whole broccoli or a mixture of one or more parts including flowers, stems, sprouts, etc.

[0029] In the present invention, broccoli / broccoli sprout exosomes extracted from broccoli / broccoli sprouts are 1 × 10 per unit volume of 1 mL of the composition. 6 ~1×10 13 pieces, preferably 1×10 7 ~1×10 13 It may be contained in the amount of individual particles.

[0030] Broccoli / broccoli sprout exosomes themselves have anti-aging and antioxidant functions similar to superoxide dismutase (SOD), which promote the decomposition of active oxygen present outside the cells and active oxygen accumulated inside the cells, and show effects such as promoting collagen production, promoting hyaluronic acid production, inhibiting melanin synthesis, and inhibiting the production of inflammatory substances.

[0031] In addition, the supply of high-quality protein is closely related to increased skin moisture and skin health, and the broccoli exosomes of the present invention contain protein at a high concentration of 10 to 1,000 μg / mL, preferably 100 to 1,000 μg / mL, and can effectively deliver plant protein to the skin.

[0032] In one embodiment, the antioxidant cosmetic composition containing the broccoli / broccoli sprout exosomes of the present invention as an active ingredient may be provided in a dosage form selected from the group consisting of an aqueous solution, a suspension, an emulsion, a cream, a frozen solution, a spray-dried powder, and a freeze-dried powder.

[0033] Another aspect of the present invention provides a cosmetic comprising an antioxidant cosmetic composition containing broccoli / broccoli sprout exosomes as an active ingredient.

[0034] The cosmetic product may be formulated with general ingredients typically used in the manufacture of skin cosmetics, such as oils, water, surfactants, moisturizers, lower alcohols, thickeners, chelating agents, pigments, preservatives, fragrances, etc., as necessary. In addition, the cosmetic product may be manufactured in a dosage form selected from the group consisting of mist, serum, nutrient lotion, softening lotion, softening lotion, emulsion, skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisture lotion, nutrient lotion, massage cream, nutrient cream, moisture cream, hand cream, foundation, powder, makeup base, essence, nutrient essence, pack, soap, cleansing foam, cleansing lotion, cleansing cream, body lotion, body cleanser, face wash, treatment, beauty serum, beauty pack, ointment, gel, liniment, liquid, patch, spray, bath additive, sunscreen, sun oil, and hair product, and may contain various conventional carriers and additives well known in the art in accordance with these dosage forms.

[0035] Such a cosmetic product of the present invention may be a functional cosmetic product having one or more efficacies selected from the group consisting of anti-aging, skin whitening, wrinkle improvement, and skin cell regeneration.

[0036] Yet another aspect of the present invention provides a method for producing an anti-aging and antioxidant cosmetic composition containing the broccoli / broccoli sprout exosomes as an active ingredient, the method comprising the steps of crushing broccoli and broccoli sprouts, centrifuging the mixture, and performing tangential flow filtration (TFF).

[0037] The process of crushing the raw plant material may include a process of mechanically crushing a mixture of the raw plant material and a buffer solution in a weight ratio of 1:1 to 1:10, which may be performed by mechanically crushing the plant mixture with a blade rotating at a speed of 10 to 1,000 rpm.

[0038] The centrifugation method may be selected from low-speed centrifugation, high-speed centrifugation, ultracentrifugation, and a combination thereof, and preferably includes ultracentrifugation. Low-speed centrifugation is performed at a speed of 6,000 rpm (6,000×g) or less and is mainly used for centrifuging samples that easily precipitate, such as cells and nuclei, while high-speed centrifugation refers to a centrifugation method with a maximum speed of about 20,000 to 25,000 rpm (60,000×g), and ultracentrifugation refers to a centrifugation method with a maximum speed of about 40,000 to 80,000 rpm (600,000×g).

[0039] The tangential flow filtration (TFF) is a filtration method in which a solution flows perpendicular to a filter membrane to filter out small impurities and separate large exosomes. Compared to conventional filtration methods, it is possible to minimize the adsorption of exosomes to the pores of the filter or the clogging of the membrane pores, and therefore it is easy to apply to process scale-up and cGMP (cosmetic good manufacturing procedure) processes. The tangential flow filtration may be at least one selected from the group consisting of hollow fiber TFF and membrane TFF capable of performing ultrafiltration, and preferably a TFF filter having a molecular weight cutoff (MWCO) of 100,000 Da to 500,000 Da.

[0040] As described above, the broccoli / broccoli sprout exosomes according to the present invention are isolated from broccoli and broccoli sprouts through mechanical disruption, stepwise ultracentrifugation and tangential flow filtration, and these exosomes contain large amounts of polyphenols having anti-aging and antioxidant functions.

[0041] [Effects of the Invention] Broccoli exosomes have a lipid bilayer structure and are absorbed into skin cells. The polyphenol components of absorbed broccoli exosomes remove active oxygen in skin cells and stimulate the antioxidant signaling mechanism, providing a variety of functional cosmetic applications, including whitening, wrinkle reduction, anti-aging, skin barrier improvement, inflammation relief, and skin cell activation. In addition, the excellent stability of broccoli exosomes allows them to be used in functional cosmetics in a variety of forms, including aqueous solutions, frozen solutions, and freeze-dried powders.

[0042] BRIEF DESCRIPTION OF THE DRAWINGS [Figure 1] A process diagram showing the plant exosome isolation method.

[0043] [Figure 2] Morphological analysis (TEM) results of broccoli sprout exosomes.

[0044] [Figure 3] Results of characterization of broccoli sprout exosomes (NTA and protein assay).

[0045] [Figure 4] Analysis results of the polyphenol content and copper reducing power of broccoli sprout exosomes.

[0046] [Figure 5] Evaluation results of the melanin production inhibitory ability of broccoli sprout exosomes.

[0047] [Figure 6] Results of evaluating the melanin production inhibitory ability in a cell model in which melanin production was promoted by a-MSH from broccoli sprout exosomes.

[0048] [Figure 7] Evaluation results of the collagen production ability of broccoli sprout exosomes.

[0049] [Figure 8] Evaluation results of the hyaluronic acid production ability of broccoli sprout exosomes.

[0050] [Figure 9] Results of evaluation of the NO production inhibitory ability of broccoli sprout exosomes.

[0051] [Figure 10] Results of evaluation of the cytotoxicity of broccoli sprout exosomes against human fibroblasts (HDF), B16F10 melanoma, and mouse macrophages (RAW 264.7).

[0052] [Mode for carrying out the invention] The present invention will be described in more detail below with reference to examples. It will be obvious to those skilled in the art to which the present invention pertains that these examples are merely for the purpose of explaining the present invention in more detail and are not intended to limit the scope of the present invention.

[0053] <Example> <Example 1> Isolation of plant exosomes Broccoli, broccoli sprouts, aloe vera skin, and spinach were used as raw materials, and plant exosomes were isolated from them using ultracentrifugation and tangential flow filtration (TFF) methods. Specifically, the plant materials were thoroughly washed and crushed, and then put into a blender and crushed with phosphate-buffered saline (PBS) at a weight ratio of 1:1 (w / w) to 1:10 (w / w). The mixture was centrifuged at 1,000 x g for 10 minutes to collect the supernatant, which was then centrifuged at 2,000 x g for 20 minutes, 3,000 x g for 30 minutes, and 10,000 x g for 60 minutes. The supernatant was then ultracentrifuged at 100,000 x g for 70 minutes at 4°C, and vacuoles were removed through this process. Finally, the supernatant was discarded, and the remaining pellet was suspended and exosomes were isolated through the TFF system. Specifically, the exosome-containing solution was concentrated by filtering the exosomes using a filter with a molecular weight cutoff of 100 to 500 kDa in the TFF system to remove other impurity particles smaller than the pores of the filter. The isolated exosomes were frozen and stored at -70°C or below until use (Figure 1).

[0054] <Example 2> Evaluation of exosome protein content The protein content of the isolated broccoli sprout exosomes, aloe exosomes, and spinach exosomes was measured using a Micro BCA® Protein Assay Kit manufactured by Thermo Scientific. The samples used were those immediately after isolation and exosome samples stored at -70°C for one month and then thawed. Specifically, 1.0 mL of each standard and sample was placed in a test tube using a pipette, and 1.0 mL of WR was added to each tube and mixed. The tubes were covered and incubated in a water bath at 37°C for 30 minutes, and then cooled at room temperature. With the spectrophotometer set to 562 nm, the instrument was zeroed in a cuvette filled with distilled water, and the absorbance of the samples was measured within 10 minutes. Protein standards were measured against the total protein concentration, and standard curves were prepared by floating the average blank-corrected 562 nm readings for each standard. The protein content of the samples was measured using the standard curve, and the results are shown in Table 1 below.

[0055] [Table 1]

[0056] As a result of the experiment, it was confirmed that broccoli sprout exosomes contained more than 100μg / mL of protein per 1mL unit volume even after thawing.Further characterization was performed using broccoli sprout exosomes that were confirmed to have a high protein content.

[0057] <Example 3> Characterization of broccoli sprout exosomes The broccoli sprout exosomes isolated in Example 1 were subjected to further exosome characterization by the following methods. The pattern of the exosomes was confirmed to be a spherical microstructure of 200 nm or less using a scanning electron microscope (SEM) (scale bar 50 nm) (Figure 2). The size of the microparticles was analyzed using nanoparticle tracking analysis (NTA) and confirmed to be 50 to 300 nm (Figure 3). Nanoparticle tracking analysis (NTA) confirmed that the number of exosomes was 1 x 10 per unit volume of 1 mL. 8 ~1×10 13The exosome concentration was confirmed (Figure 3). The protein content in the exosomes was additionally measured several times using the Thermo Scientific Micro BCA® Protein Assay Kit, and it was confirmed that the exosomes contained 10 to 1,000 μg / mL of protein per 1 mL unit volume (Figure 3).

[0058] <Example 4> Evaluation of polyphenol content in broccoli sprout exosomes The polyphenol content in the exosomes of the broccoli sprout exosomes isolated in Example 1 was evaluated by the following method. The polyphenol content in the broccoli exosomes was measured using a phenolic compound assay kit manufactured by BioVision. The measurement method followed the protocol of the kit manufacturer. 8 It was confirmed that each broccoli exosome contained 50 to 500 pmol of polyphenols (Figure 4).

[0059] <Example 5> Evaluation of antioxidant activity of broccoli sprout exosomes The polyphenol content in the exosomes of the broccoli sprout exosomes isolated in Example 1 was evaluated by the following method. The antioxidant capacity of the broccoli exosomes was evaluated using the OxiSelect (registered trademark) Total Antioxidant Capacity (TAC) Assay Kit manufactured by Cell Bio Labs. The measurement method followed the protocol of the kit manufacturer. As a result of the evaluation, it was confirmed that the broccoli sprout exosomes have the ability to reduce 10 to 100 μM of Cu(II) ions to Cu(I) ions per unit volume of 1 mL (Figure 4).

[0060] Example 6: Evaluation of the in vitro whitening efficacy of broccoli sprout exosomes In order to evaluate the antioxidant effect of the broccoli sprout exosomes isolated in Example 1, the skin whitening effect of the exosomes was evaluated as a representative example. 3 ~1×10 4The broccoli exosome-containing cell culture medium was treated in 16 skin melanoma (B16F10) cells and cultured for 3 days, after which the amount of melanin accumulated inside and outside the cells was measured. The melanin accumulated inside the cells was measured by destroying the cells with a lysis buffer and measuring the absorbance of the solution at 475 nm using a UV-Vis spectrophotometer. The melanin accumulated outside the cells was measured by collecting the cell culture medium after culturing for 3 days and measuring the absorbance of the solution at 490 nm using a UV-Vis spectrophotometer. The negative control group was a cell culture medium (GM) that had not been subjected to any treatment, and the positive control group was 0.5 mM arbutin, which is widely used to evaluate skin whitening efficacy.

[0061] Broccoli sprout exosomes are 5×10 7 At doses of 5×10 or more, melanin production was suppressed both extracellular and intracellularly. 8 It was confirmed that treatment with 100 mg of ...

[0062] The whitening efficacy of the broccoli sprout exosomes isolated in Example 1 was further evaluated using a model in which skin melanoma (B16F10) cells were treated with 100 nM α-MSH to promote melanin synthesis and secretion. After treating the cell culture medium containing α-MSH and broccoli exosomes, the cells were cultured for 3 days, and the total content of melanin accumulated inside the cells and melanin secreted outside the cells was measured using an ultraviolet-visible spectrophotometer. Protein content was quantified using the Bradford method, and the amount of melanin was converted to the amount of melanin per protein. Cell culture medium not treated with exosomes was used as a negative control group, and 0.36 mM arbutin, which is widely used to evaluate whitening efficacy, was used as a positive control group. Broccoli sprout exosomes were cultured for 3 days after treatment with α-MSH and broccoli exosomes, and the total content of melanin accumulated inside the cells and melanin secreted outside the cells was measured using an ultraviolet-visible spectrophotometer. Protein content was quantified using the Bradford method, and the amount of melanin was converted to the amount of melanin per protein. Cell culture medium not treated with exosomes was used as a negative control group, and 0.36 mM arbutin, which is widely used to evaluate whitening efficacy, was used as a positive control group. Broccoli sprout exosomes were cultured at 2 × 10 7 The above doses inhibited total melanin production (p<0.05) (Figure 6).

[0063] Example 7: Evaluation of in vitro collagen production from broccoli sprout exosomes To evaluate the efficacy of the broccoli sprout exosomes isolated in Example 1 on collagen production, human dermal fibroblasts (HDFs) were treated with exosomes to confirm the amount of collagen produced within the cells. HDFs cells were treated with cell culture medium containing broccoli exosomes and cultured for 2 days, after which they were centrifuged and the amount of procollagen was measured in the supernatant. The amount of procollagen was measured by measuring the absorbance at 450 nm using a procollagen type I C-peptide (PIP) ELISA kit. The cell culture medium (GM) that had not been subjected to any treatment was used as the negative control group, and TGF-β1, which is known to induce collagen production, was used as the positive control group. Broccoli sprout exosomes increased collagen production by 1.3 × 10 compared to the negative control group. 6 ~1×10 7 It was confirmed that the concentration of 100 mg / kg / day had a significant effect on increasing intracellular collagen production (p<0.05) (Figure 7), which is similar to the efficacy of the positive control TGF-β1.

[0064] Example 8: Evaluation of in vitro hyaluronic acid production from broccoli sprout exosomes To evaluate the efficacy of the broccoli sprout exosomes isolated in Example 1 in producing hyaluronic acid, human dermal fibroblasts (HDFs) were treated with the exosomes to confirm the amount of hyaluronic acid produced within the cells. HDFs cells were treated with cell culture fluid containing broccoli exosomes and cultured for 2 days, after which the cells were centrifuged and the amount of hyaluronic acid produced in the supernatant was measured. The amount of hyaluronic acid produced was measured by measuring the absorbance at 450 nm using a human hyaluronic acid (HA) ELISA kit. The negative control group was cell culture fluid (GM) that had not been subjected to any treatment, and the positive control group was retinoic acid, which induces the production of hyaluronic acid. Broccoli sprout exosomes increased the amount of hyaluronic acid produced within the cells by 2.5 × 10 compared to the negative control group. 6 ~1×10 7 It was confirmed that the concentration of 100 mg / mL ...

[0065] Example 9: Evaluation of broccoli sprout exosomes in vitro for inhibition of nitric oxide (NO) production To evaluate the effect of the broccoli sprout exosomes isolated in Example 1 in suppressing nitric oxide (NO) production, mouse macrophages (RAW 264.7 cells) were treated with the exosomes to confirm whether NO production was suppressed. 5 RAW 264.7 cells were cultured for 20 hours, then replaced with new cell culture medium containing 1 μg / mL lipopolysaccharide (LPS) and broccoli exosomes, and cultured for 24 hours. After 24 hours, the culture medium was reacted with the NO detection reagent for 10 minutes, and the absorbance of the solution was measured at 540 nm. The amount of NO produced was evaluated by correcting for the total protein amount, and dexamethasone 7.8 μg / mL, which has an anti-inflammatory effect, was used as a positive control.

[0066] Broccoli sprout exosomes were 2×10 7 The above confirmed the efficacy of suppressing NO production (p<0.05) (Figure 9). 7 These exosomes effectively suppressed cellular NO production, with efficacy similar to that of the positive control, dexamethasone.

[0067] <Example 10> Evaluation of cytotoxicity of broccoli sprout exosomes To confirm the cytotoxicity of the broccoli sprout exosomes isolated in Example 1, the cell viability of two types of human cells and mouse macrophages after exosome treatment was evaluated using the MTT assay.

[0068] 5 × 10 human dermal fibroblasts 4 After inoculation at a concentration of 1.5 × 10 cells / well and incubation for 24 hours, broccoli exosomes were treated at different concentrations and then incubated for another 24 hours. 4 After inoculation at a concentration of 1.8 × 10 cells / well and incubation for 24 hours, the cells were treated with broccoli exosomes at different concentrations and incubated for 72 hours. Mouse macrophages (RAW 264.7) were inoculated at a concentration of 1.8 × 10 5After inoculation at a concentration of 1000 cells / well and incubation for 20 hours, broccoli exosomes were treated at different concentrations and incubated for 24 hours. Each well was treated with MTT reagent at a concentration of 0.05% in cell culture medium and incubated for 4 hours. After removing the culture medium, DMSO was inoculated at 1 mL / well and stirred for 10 minutes, and the DMSO solution was collected and the absorbance was measured at 540 nm. The cell viability was shown as a percentage compared to the group not treated with exosomes (Figure 10). The experimental results confirmed that broccoli sprout exosomes had no cytotoxicity at all concentrations. [Brief description of the drawings]

[0069] [Figure 1] FIG. 1 is a process diagram showing a method for isolating plant exosomes. [Diagram 2] These are the results of morphological analysis (TEM) of broccoli sprout exosomes. [Diagram 3] These are the results of characterization (NTA and protein assay) of broccoli sprout exosomes. [Figure 4] 1 shows the results of an analysis of the polyphenol content and copper reducing power of broccoli sprout exosomes. [Diagram 5] These are the results of evaluating the ability of broccoli sprout exosomes to inhibit melanin production. [Figure 6] These are the results of evaluating the melanin production inhibitory ability in a cell model in which melanin production was promoted by a-MSH from broccoli sprout exosomes. [Figure 7] These are the results of evaluating the collagen production ability of broccoli sprout exosomes. [Figure 8] These are the results of evaluating the hyaluronic acid production ability of broccoli sprout exosomes. [Figure 9] These are the results of evaluating the ability of broccoli sprout exosomes to inhibit NO production. [Figure 10] These are the results of evaluating the cytotoxicity of broccoli sprout exosomes against human fibroblasts (HDF), B16F10 melanoma, and mouse macrophages (RAW 264.7).

Claims

1. A skin whitening agent or hyaluronic acid generating agent containing, as an active ingredient, broccoli exosomes isolated from broccoli raw material including broccoli sprouts.

2. The skin whitening agent or hyaluronic acid generating agent according to claim 1 , wherein the broccoli raw material further comprises at least one selected from the group consisting of broccoli flowers and broccoli stems.

3. The broccoli exosomes are added at a concentration of 10 per 1 mL of the skin whitening agent or hyaluronic acid generating agent. ６ pieces ~ 10 １３ The skin whitening agent or hyaluronic acid generating agent according to claim 1, which is contained in an amount of 100 mg / kg or more.

4. The skin whitening agent or hyaluronic acid generator according to claim 1, wherein the skin whitening agent or hyaluronic acid generator is in a dosage form selected from the group consisting of an aqueous solution, a suspension, an emulsion, a cream, a frozen solution, a spray-dried powder, and a freeze-dried powder.

5. The skin whitening agent or hyaluronic acid generating agent according to claim 1 , wherein the broccoli exosomes contain polyphenols.

6. The skin whitening agent or hyaluronic acid generating agent according to claim 1 , further having a collagen generating application, an anti-aging application or an antioxidant application.

7. Use of a skin whitening agent or hyaluronic acid generator described in any one of claims 1 to 6 in the manufacture of a cosmetic product.

8. The use of a skin whitening agent or a hyaluronic acid generator according to claim 7, wherein the cosmetic product is in a dosage form selected from the group consisting of mist, serum, nutrient lotion, softening lotion, softening water, emulsion, skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisture lotion, nutrient lotion, massage cream, nutrient cream, moisture cream, hand cream, foundation, powder, makeup base, essence, nutrient essence, pack, soap, cleansing foam, cleansing lotion, cleansing cream, body lotion, body cleanser, facial wash, treatment, beauty serum, beauty pack, ointment, gel, liniment, liquid, patch, spray, bath additive, sunscreen, sun oil and hair product.

9. The use of a skin whitening agent or a hyaluronic acid generating agent according to claim 7, wherein the cosmetic is a functional cosmetic having one or more efficacies selected from the group consisting of skin whitening and hyaluronic acid generation.

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