KR102277721B1 - Manufacturing method of tissue cultured Panax ginseng C.A. Meyer adventitious root extract using a novel biological network mimic method and cosmetic composition containing the same - Google Patents

Abstract

Research on useful plant ingredients has been mainly conducted to find one active ingredient under the assumption that one or several active ingredients in plants determine a physiological activity for centuries. It has been recently found that useful ingredients of plants result from complicated actions among fat-soluble substances found in cell membranes as low-molecular substances present in plants, polar substances from water-soluble parts of cells, acidic/basic ions, and stable, oxidized structures, and the like. However, a conventional method for extracting useful ingredients from plants has a problem in that an extract reflecting a network between substances such as in cells is not prepared. The present invention relates to a novel biological network mimic extraction method of plant useful ingredients capable of complexly reflecting plant metabolites such as a network between substances in a cell, and the plant useful ingredients extracted by the extraction method of the present invention are expected to be actively used in medical, bio, cosmetic, and food fields.

Description

A method for preparing a ginseng extract using a novel bionetwork mimic extraction method and a cosmetic composition comprising the same {Manufacturing method of tissue cultured Panax ginseng C.A. Meyer adventitious root extract using a novel biological network mimic method and cosmetic composition containing the same}

The present invention relates to a novel biological network mimic extraction method of plant useful components, and more particularly, to a useful component extraction method that complexly reflects plant metabolites such as a network between substances in a cell.

Efforts have been made to extract useful ingredients from plants and use them in cosmetic compositions, food compositions, or pharmaceutical compositions, but research on plant useful ingredients has been based on the assumption that one or several active ingredients in plants determine the physiological activity for centuries. It was mainly done in the direction of finding one active ingredient under However, according to a recent research report, a meta-analysis study that synthesizes and analyzes the results of clinical trial studies on vitamins and antioxidants known to be effective in the human body reveals that vitamins alone or antioxidants alone cannot produce biological effects. On the other hand, the meta-analysis of clinical trials in which vegetables or fruits were ingested showed efficacy, which is interpreted because vitamins and antioxidants do not act alone in the body, but form a network in the form of a complex. According to other studies, vitamin complexes do not show effectiveness, but it is analyzed that vitamin complexes show positive effects of vitamin complexes only when other nutrients of fruits and vegetables work together. Metabolomics, which emerged as such a concept, is a low molecular weight substance in which useful components of plants exist in plants, fat-soluble substances found in cell membranes, polar substances from water-soluble parts of cells, acidic/basic ions, and stable structures. It is shown that the result of a complex action of hyperoxidized structures, etc. Therefore, in order to obtain the expected effect from the composition containing the plant useful component, the total data of metabolites obtained from the plant should be reflected as the useful component.

On the other hand, the conventional method for extracting useful components from plants is single solvent extraction or single-system mixed solvent extraction in which two or three solvents are mixed, so that the effective components of the raw materials are not extracted integrally, and simple fraction extraction is performed by solubility in the extraction solvent. There was a problem in that an extract reflecting the network between substances as in cells was not prepared. In order to extract useful components as metabolites from plants, a new extraction technology is required to simultaneously extract the water-soluble and oil-soluble effective components of raw materials while creating a membrane structure so that the network between substances can be maintained similar to that in the living body.

Therefore, the inventors of the present invention have completed the present invention after continuous efforts to solve the problems raised above. The present invention relates to a novel biological network imitation extraction method of plant useful components capable of complexly reflecting plant metabolites such as a network between substances in cells, and the plant useful components extracted by the extraction method of the present invention are medical, bio, cosmetic , and is expected to be actively used in the food field.

The present invention relates to a novel biological network mimic extraction method of plant useful components.

To this end, the present invention provides a novel extraction that creates a membrane structure so that a network between substances can be maintained similarly to that in a living body while simultaneously extracting water-soluble and oil-soluble effective components of plant raw materials.

In addition, it provides a cosmetic composition comprising the useful component extracted by the above extraction method.

In addition, it provides a food composition comprising the useful component extracted by the extraction method.

In addition, it provides a medical composition comprising the useful component extracted by the extraction method.

However, the technical task to be achieved by the present invention is not limited to the tasks mentioned above, and other tasks not mentioned will be clearly understood by those of ordinary skill in the art from the following description.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS Various specific details are set forth below, such as specific forms, compositions and processes, and the like, for a thorough understanding of the present invention. However, certain embodiments may be practiced without one or more of these specific details, or in conjunction with other known methods and forms. In other instances, well-known processes and manufacturing techniques have not been described in specific detail in order not to unnecessarily obscure the present invention. Reference throughout this specification to “one embodiment” or “an embodiment” means that a particular feature, form, composition, or characteristic described in connection with the embodiment is included in one or more embodiments of the invention. Thus, references to "in one embodiment" or "an embodiment" in various places throughout this specification do not necessarily refer to the same embodiment of the invention. Additionally, the particular features, forms, compositions, or properties may be combined in any suitable manner in one or more embodiments.

Unless otherwise defined in the present invention, all scientific and technical terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

As used herein, “metabolomics” refers to the systematic and systematic analysis of the profiles of small molecule metabolites generated through specific biological change processes, including genomics or proteomics, which study the presence or absence of changes in genes or proteins. It means a discipline that studies comprehensively. Derived from this, “metabolite” is a concept that complexly encompasses intermediates or final products of metabolic processes as small molecules. More specifically, in general, a metabolite is a quantifiable small molecule that best represents the phenotype of a living body, and the term “metabolite” is a low molecular weight (100 to 1,000 MW) molecule that exists in a cell, tissue, or living body. It refers to an aggregation (metabolic intermediates, hormones, other signaling molecules, and secondary metabolites, etc.) and forms a large network of metabolic reactions.

As used herein, the term “useful ingredient” refers to a physiologically active ingredient extracted from a plant, preferably a low-molecular substance present in a plant, which is found in the cell membrane and is found in fat-soluble substances, polar substances from water-soluble parts of the cell, acidic/basic It refers to a component (hereinafter referred to as a 'metabolite useful component') including ions, and a stable structure and an oxidized structure.

As used herein, the term "metabolite extraction method" refers to a method of extracting the metabolite useful component from a plant. The conventional method for extracting useful components of plants is a method of separating and extracting the active ingredients of natural products by solubility of water or organic solvents, and in this extraction method, there is no choice but to have limitations in extracting while maintaining the biological network structure of natural products.

Therefore, the metabolite extraction method of the present invention is intended to improve the disadvantages of the conventional single solvent extraction or single-system mixed solvent extraction method in which two or more solvents and a surfactant are simply mixed, and according to the solubility of biocomponents in a single-phase solvent system, It is not a method of fractional extraction, but a solvent system in which a fine dual phase (oil-soluble solvent such as liposome is finely fractionated) is used. This is a method in which oil-soluble water-soluble components are mixed and extracted similarly to a living body, not a fractional extract according to solubility, which is a disadvantage of the conventional extraction method. The advantage of this extraction method is that since a network of oil-soluble and water-soluble active ingredients is formed in the extract, high efficacy can be exhibited similarly to in vivo. More specifically, in the metabolite extraction method of the present invention, a fat-soluble part composed of phospholipids, lipids having 12 or more carbon atoms, a nonionic surfactant, etc. and a water-soluble part composed of alcohols having 2 to 5 carbon atoms, purified water, etc., and a plant after mixing It is characterized in that the emulsion of the liposome or membrane structure is made while the phospholipid and the surfactant are mixed with the biocomponent during the extraction process by stirring, ultrasonication, or fine grinding. This is to allow the biocomponents to be extracted into a network structure similar to a cell. During the extraction process, each solvent, phospholipid and surfactant are self-rearranged by micro-mixing. Accordingly, this is a method of simultaneously forming and extracting the membrane structure. The water-soluble active ingredient is extracted from the aqueous phase of the membrane structure, and the oil-soluble active ingredient is the membrane. By simultaneously extracting the lipid phase of the structure, the network and vesicle shape between the active ingredients can be maintained similar to that of living cells.

The "metabolite extraction method" of the present invention has a technical similarity in that it applies the conventional liposome production technology or nanoemulsion production technology, but the conventional liposome application technology simply collects the extracted single component or complex component into the liposome. Compared to the technique of increasing the usability, the present invention has technological advances in that it is possible to obtain an extract that mimics a biological network at the same time as extraction in the extraction step.

The extraction method includes adding and stirring phospholipids, surfactants, lipids other than phospholipids, water-soluble solvents and water to the extraction raw material.

In the extraction method of the present invention, the step comprises the steps of preparing an oil phase by mixing lipids other than the phospholipids, surfactants and phospholipids; preparing an aqueous phase by mixing the water-soluble solvent and water; and adding the oil phase part and the aqueous phase part to the extraction raw material and stirring.

In the extraction method of the present invention, the surfactant is preferably at least one selected from the group consisting of a span type surfactant, a twin type surfactant and a nonionic surfactant.

In the extraction method of the present invention, the lipid other than the phospholipid is preferably a lipid having 12 or more carbon atoms.

In the extraction method of the present invention, the lipid other than the phospholipid is preferably at least one selected from the group consisting of fatty acid esters and vegetable oils.

In the extraction method of the present invention, the water-soluble solvent is preferably an alcohol having 2 to 6 carbon atoms.

The present invention provides a metabolite extraction method, and a metabolite useful component extracted from a plant by this method.

The above plants are not limited to the species, and may include any one or both of the above-ground part or the underground part. The above-ground part is a part that is exposed above the ground (soil) among the parts constituting the plant. In general, elements belonging to the above-ground part include a stem, leaf, flower, bark, fruit, etc., but is not limited thereto. The underground part means a part below the ground (soil) among parts constituting the plant. In general, an element belonging to the underground part means a root, and the root includes stored roots such as radish, carrot, ginseng, arrowroot, sweet potato, and the like. Even if it exists under the ground or stems such as bamboo shoots, taro, lotus root, etc., it may be included in the underground part.

In the present invention, the plant used for the metabolite useful component is ginseng, and collectively refers to ginseng and wild ginseng harvested or cultivated in nature. Ginseng, wild ginseng, wild ginseng, wild ginseng, wild ginseng, sprout ginseng, dried ginseng, fresh ginseng, white ginseng, red ginseng, direct ginseng, gok ginseng, bangok ginseng, go ginseng, seonhwa ginseng, hwansam, ginseng, ginseng, ginseng, cheonsam, cheonjong ginseng, jijong ginseng, cheonjong ginseng , and wild ginseng, but is not limited thereto.

The present invention also provides a cosmetic composition comprising the metabolite useful component.

The cosmetic composition includes lotion, nutritional lotion, nutritional essence, massage cream, cosmetic bath additive, body lotion, body milk, bath oil, baby oil, baby powder, shower gel, shower cream, sunscreen lotion, sunscreen cream, suntan cream , skin lotion, skin cream, sunscreen cosmetic, cleansing milk, depilatory {cosmetic}, face and body lotion, face and body cream, skin whitening cream, hand lotion, hair lotion, cosmetic cream, jasmine oil, bath soap , water soap, beauty soap, shampoo, hand sanitizer (hand cleaner), medicated soap {non-medical use}, cream soap, facial wash, hair rinse, cosmetic soap, tooth whitening gel, toothpaste, and the like. To this end, the composition of the present invention may further include an appropriate carrier, excipient or diluent commonly used in the preparation of cosmetic compositions. Carriers, excipients or diluents that may be further added to the cosmetic composition of the present invention include purified water, oil, wax, fatty acid, fatty alcohol, fatty acid ester, surfactant, humectant, thickener, antioxidant, viscosity stabilizer, kill rating agents, buffers, lower alcohols, and the like, but are not limited thereto. In addition, if necessary, it may include a whitening agent, a moisturizer, a vitamin, a sunscreen, a perfume, a dye, an antibiotic, an antibacterial agent, an antifungal agent. Hydrogenated vegetable oil, castor oil, cottonseed oil, olive oil, palm oil, jojoba oil, and avocado oil may be used as the oil. As the wax, beeswax, spermaceti, carnauba, candelilla, montan, ceresin, liquid paraffin, and lanolin may be used. can be used As the fatty acid, stearic acid, linoleic acid, linolenic acid, and oleic acid may be used, and as the fatty acid alcohol, cetyl alcohol, octyldodecanol, oleyl alcohol, panthenol, lanolin alcohol, stearyl alcohol, and hexadecanol may be used. And as the fatty acid ester, isopropyl myristate, isopropyl palmitate, butyl stearate may be used. As the surfactant, cationic surfactants, anionic surfactants and nonionic surfactants known in the art can be used, and surfactants derived from natural products are preferred as much as possible. In addition, it may include a desiccant, a thickener, an antioxidant, etc. widely known in the cosmetic field, and the types and amounts thereof are as known in the art.

The present invention also provides a food composition comprising the metabolite useful component.

The food composition may be prepared in the form of various foods, for example, beverages, gum, tea, vitamin complexes, powders, granules, tablets, capsules, confectionery, rice cakes, bread, and the like. Since the food composition of the present invention is improved from the existing food intake with little toxicity and side effects, it can be safely used even when taken for a long period of time for the purpose of prevention. When the composition of the present invention is included in the food composition, the amount may be added in a proportion of 0.1 to 100% of the total weight. Here, when the food composition is prepared in the form of a beverage, there is no particular limitation other than containing the food composition in the indicated ratio, and it may contain various flavoring agents or natural carbohydrates as additional ingredients, as in a conventional beverage. That is, as natural carbohydrates, monosaccharides such as glucose, disaccharides such as fructose, polysaccharides such as sucrose, and common sugars such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol are included. can do. Examples of the flavoring agent include natural flavoring agents (taumatin, stevia extract (eg, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.). Others of the present invention of various nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic and natural flavoring agents, coloring agents, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH regulators, It may contain stabilizer, preservative, glycerin, alcohol, carbonation agent used in carbonated beverage, etc. These components can be used independently or in combination.The proportion of these additives is usually per 100 parts by weight of the composition of the present invention. It is generally selected in the range of 0.1 to 100 parts by weight, but is not limited thereto.

The present invention also provides a pharmaceutical composition comprising the metabolite useful component.

The pharmaceutical composition is administered for the treatment of a disease, including the activity of useful ingredients, age, weight, general health, sex, formula, administration time, administration route, excretion rate, drug formulation and the severity of the specific disease to be prevented or treated It may vary depending on various factors, and the dosage of the pharmaceutical composition may vary depending on the patient's condition, weight, degree of disease, drug form, administration route and period, but may be appropriately selected by those skilled in the art. The pharmaceutical composition according to the present invention may be formulated as pills, dragees, capsules, solutions, gels, syrups, slurries, and suspensions. In addition, the pharmaceutical composition according to the present invention contains the useful component of the present invention as an active ingredient in an amount of 0.1 to 50% by weight based on the total weight of the composition. It may include a compound involved in the treatment of a desired disease and a pharmaceutically acceptable carrier, excipient or diluent. For example, carriers, excipients and diluents that may be included in the composition of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate. , calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil. .

In one embodiment of the present invention, (a) preparing a fat-soluble part by mixing a phospholipid, a surfactant, and a lipid; (b) mixing a water-soluble solvent and purified water to prepare a water-soluble part; And, (c) adding the (a) and (b) to the plant and stirring; provides a method for producing a vesicle in which useful components of plant metabolites are collected, the plant comprising any one of an above-ground part or an underground part It provides a method for producing a vesicle in which a plant metabolite useful component is captured, the metabolite useful component is a low-molecular substance present in a plant, including fat-soluble substances found in cell membranes, and polarity from water-soluble parts of cells It provides a method for producing a vesicle in which a plant metabolite useful component is collected, which is a component comprising a complex of substances, acid/basic ions, and a stable structure and an oxidized structure, the method comprising: a metabolite from a plant Provided is a method for producing a vesicle in which useful components of plant metabolites are captured at the same time as useful components are extracted and collected in the vesicles.

Specifically, the phospholipids in step (a) are hydrogenated from the group consisting of phosphatidylcholine, phosphatidylserine, phosphatidylethanolamine, phosphatidylinositol, lecithin, and their hydrogenated substances. It provides a method for producing a vesicle in which useful components of plant metabolites that are at least one selected type are captured.

In addition, the surfactant in step (a) provides a method for producing a vesicle in which a plant metabolite useful component is captured, which is a span-type surfactant, a tween-type surfactant, or a nonionic surfactant, and the span-type surfactant may be sorbitan laurate, sorbitan palmirate, sorbitan stearate, and sorbitan oleate, and the tween-type surfactant is polysorbate (polysorbate), polysorbate 20 (polysorbate 20), polysorbate 60 (polysorbate 60), and polysorbate 80 (polysorbate 80), wherein the non-ionic surfactant is PEG-8 caprylic/capric glycerin. Ride (PEG-8 caprylic / capric glycerides), poloxamer 188 (poloxamer 188), and poloxamer 407 (poloxamer 407), which may be at least one selected from the group of surfactants, vegetable metabolite useful components are captured Provided is a method for producing a cleavage.

In addition, the lipid in step (a) provides a method for producing a vesicle in which a plant metabolite useful component is captured, which is a lipid having 12 or more carbon atoms, and the lipid is a fatty acid ester, or a vegetable oil. It provides a method for producing a vesicle in which a useful component of a phosphorus plant metabolite is captured, wherein the fatty acid ester is a group consisting of cetyl caprate, cetyl palmitate, and cetyl ethylhexanoate. It provides a method for producing vesicles in which useful components of plant metabolites are collected, wherein the vegetable oil is argan oil, avocado oil, grape seed oil, macadamia It provides a method for producing a vesicle in which the useful component of a plant metabolite is at least one selected from the group consisting of nut oil (macadamia nut oil), and olive oil.

In addition, the water-soluble solvent in step (b) provides a method for producing a vesicle in which useful components of plant metabolites are collected, which is an alcohol having 2 to 5 carbon atoms, and the water-soluble solvent is ethanol (ethyl alcohol), propanol (propyl alcohol) ), isopropyl alcohol, propylene glycol, isopropylene glycol, dipropylene glycol, butylene glycol, ethoxydiol Glycol (ethoxydiglycol), hexanediol (hexanediol), glycerin (glycerin), and at least one selected from the group consisting of methylpyrrolidone (methylpyrrolidone) to provide a method for producing a vesicle in which useful components of plant metabolites are collected do.

In another embodiment of the present invention, there is provided a vesicle in which the plant metabolite useful component is captured, prepared by the method for producing the vesicle in which the plant metabolite useful component is captured.

In another embodiment of the present invention, there is provided a cosmetic composition comprising a vesicle in which the plant metabolite useful ingredient is captured, prepared by the method for producing a vesicle in which the plant metabolite useful ingredient is captured, and the cosmetic composition is a skin antioxidant , wrinkle improvement, or provides a cosmetic composition that has an antibacterial effect.

In another embodiment of the present invention, there is provided a food composition comprising a vesicle in which the plant metabolite useful ingredient is captured, prepared by the method for producing a vesicle in which the plant metabolite useful ingredient is captured, and the food composition is a skin antioxidant , Wrinkle improvement, or provides a food composition that has an antibacterial effect.

The conventional method for extracting useful components from plants is single solvent extraction or single-system mixed solvent extraction in which two or three solvents are mixed, so that the effective ingredients of the raw materials are not extracted integrally, and only fractional extraction is performed by solubility in the extraction solvent. Therefore, there was a problem that the extract reflecting the network between substances as in the cell was not prepared.

The present invention relates to a novel bio-network mimic extraction method of plant useful components. When the extraction method of the present invention is used, fat-soluble substances found in cell membranes as low-molecular substances present in plants, and polar substances derived from water-soluble parts of cells It is possible to obtain a metabolite useful component including complexes, acid/basic ions, and stable and oxidized structures.

1 is a result confirming the pro-collagen type 1 synthesis promoting effect of Comparative Examples 1 to 5, and Preparation Example 1 according to an embodiment of the present invention.

Hereinafter, the present invention will be described in more detail through examples. These examples are only for illustrating the present invention in more detail, and it will be apparent to those of ordinary skill in the art that the scope of the present invention is not limited by these examples according to the gist of the present invention. .

The metabolite extraction method of the present invention is a method in which extraction of useful components from a plant and formation of a membrane structure proceed simultaneously. Specifically, the water-soluble active ingredient is extracted from the aqueous phase of the membrane structure, and the oil-soluble active ingredient is simultaneously extracted from the lipid phase of the membrane structure. It is a method that allows the network and vesicle shape between the active ingredients to be maintained similar to that of living cells during extraction. That is, by adding and stirring phospholipids, surfactants, lipids other than phospholipids, water-soluble solvents and water to the extraction raw materials, emulsification of the extraction medium is performed and useful components are extracted, so that the water-soluble and fat-soluble components of the extraction raw materials are extracted together. In addition, it is characterized in that, by generating micro-dual-phase particles in the form of liposomes, for example, components present in the membrane of cells or organelles can be efficiently extracted. According to the extraction method of the present invention, it is possible to extract the water-soluble component, the fat-soluble component of the extraction raw material, and the components present in the membrane together, and these components are stably (for example, a composition obtained by extraction) in the extraction composition (composition obtained by extraction). The water-soluble component may be present in the aqueous phase of the extract composition in an emulsion state, the oil-soluble component may be present in the oil phase, and the component present in the membrane may be present between the phospholipid bilayers of the microbiphase particles).

Accordingly, water-soluble components, fat-soluble components, and membrane components related to the exertion of physiological activity (for example, antioxidant effect) can be stably present in the extract composition, so excellent physiological activity similar to that exhibited in vivo through the network of these components is achieved. can be exerted through the extract composition.

To this end, a fat-soluble part and a water-soluble part of the extraction solvent system are separately prepared. More specifically, the fat-soluble part is prepared by mixing 0.1 to 30% phospholipid, 0.1 to 30% surfactant, and 0.1 to 70% lipid, followed by stirring at 20 to 100°C. The phospholipid can be used by selecting one or more from the group consisting of phosphatidylcholine, phosphatidylserine, phosphatidylethanolamine, phosphatidylinositol, lecithin, and hydrogenated substances thereof. . The surfactant is a span-type surfactant such as sorbitan laurate, sorbitan palmirate, sorbitan stearate, sorbitan oleate, polysorbate ( tween surfactants such as polysorbate), polysorbate 20, polysorbate 60, and polysorbate 80, PEG-8 caprylic/capric glyceride (PEG-8) At least one selected from the group consisting of nonionic surfactants such as caprylic/capric glycerides), poloxamer 188, and poloxamer 407 can be used. The lipid is a lipid other than a phospholipid, preferably a lipid having 12 or more carbon atoms. Preferably, at least one selected from the group consisting of fatty acid esters and vegetable oils, fatty acid esters such as cetyl caprate, cetyl palmitate, cetyl ethylhexanoate, etc. ester), argan oil, avocado oil, grape seed oil, macadamia nut oil, vegetable oil such as olive oil, etc. More than one can be selected. The water-soluble part is prepared by mixing purified water with 0.1 to 70% of a water-soluble solvent and stirring at 20 to 100°C. The water-soluble solvent is preferably an alcohol having 2 to 6 carbon atoms, ethanol (ethyl alcohol), propanol (propyl alcohol), isopropanol (isopropyl alcohol), propylene glycol (propylene glycol), isopropylene glycol (isopropylene glycol) ), dipropylene glycol, butylene glycol, ethoxydiglycol, hexanediol, glycerin, methylpyrrolidone It can be used by selecting one or more types from the group consisting of.

In the extraction method of the present invention, the addition of phospholipids, surfactants, lipids other than phospholipids, water-soluble solvents and water to the extraction raw materials is preferably performed within 1 hour. That is, it is preferable that the time it takes to add all of the five extraction medium components (phospholipids, surfactants, lipids other than phospholipids, water-soluble solvents, and water) to the extraction raw material is less than 1 hour. For example, when each of the five extraction medium components is added in order, the time from the start of the first input of the extraction medium component to the last time the input of the fifth extraction medium component ends is 1 It is preferable to do it within an hour. More preferably, this time is set to 30 minutes or less, more preferably 20 minutes, more preferably 15 minutes, still more preferably 10 minutes, still more preferably 5 minutes, still more preferably within 1 minute, further Preferably at the same time.

In the extraction method of the present invention, the amounts of phospholipids, surfactants, lipids other than phospholipids, water-soluble solvents and water are preferably 0.1 to 30 phospholipids based on the combined weight of all five extraction medium components (the amount of total extraction medium components). % by weight, surfactant 0.1 to 30% by weight, lipids other than phospholipids 0.1 to 70% by weight, water-soluble solvent 0.1 to 70% by weight, the remainder being water. More preferably, based on the total weight of the five extraction medium components, 0.5 to 20% by weight of phospholipids, 2 to 30% by weight of surfactant, 0.1 to 20% by weight of lipids other than phospholipids, 15 to 50% by weight of water-soluble solvent , do the rest with water. More preferably, 2 to 6 wt% of phospholipids, 3 to 20 wt% of surfactants, 1 to 5 wt% of lipids other than phospholipids, and 15 to 40 wt% of water-soluble solvents based on the total weight of the five extraction medium components , do the rest with water.

In the extraction method of the present invention, a method in which phospholipids, surfactants, lipids other than phospholipids, water-soluble solvents and water are divided into an oil phase and an aqueous phase and separately prepared, and then added to the extraction raw material may be used. For example, a method of preparing an oil phase by mixing phospholipids, surfactants, and lipids other than phospholipids, preparing an aqueous phase by mixing a water-soluble solvent and water, and then adding the oil phase and aqueous phase to the extraction raw material may be used. According to this, it is possible to increase the expected effect according to the extraction method of the present invention as described above.

At this time, the addition of the oil phase part and the aqueous phase part to the extraction raw material is preferably made within 1 hour. That is, it is preferable that the time it takes to add both the oil phase part and the aqueous phase part to the extraction raw material is within 1 hour. For example, when the oil phase part and the water phase part are added in order, the time from the start of the input of the first receiving phase (or the oil phase) to the end of the input of the next oil phase part (or the water phase) is less than 1 hour it is preferable More preferably, this time is set to 30 minutes or less, more preferably 20 minutes, more preferably 15 minutes, still more preferably 10 minutes, still more preferably 5 minutes, still more preferably within 1 minute, further Preferably at the same time.

In the extraction method of the present invention, the ratio of the extraction raw material to the extraction medium is preferably 1 to 1000 parts by weight of the extraction medium (based on the weight of the sum of the five extraction medium components) based on 1 part by weight of the extraction raw material, more preferably is 2 to 100 parts by weight, more preferably 5 to 50 parts by weight.

In the extraction method of the present invention, stirring may be performed using a stirrer. The type of stirrer is not particularly limited, and for example, a general stirrer, a double helix stirrer, a high-speed emulsifier, a homogenizer, a mixer, or an ultrasonic homogenizer may be used.

The temperature applied during stirring in the extraction method of the present invention is preferably 10 to 80 ° C. based on the central temperature of the mixture, that is, the mixture of the extraction raw material, phospholipids, surfactants, lipids other than phospholipids, water-soluble solvents and water, and more Preferably it is 20-80 degreeC, More preferably, it is set as 20-60 degreeC. The stirring time is preferably 10 minutes to 30 days, and more preferably 30 minutes to 72 hours. In the case of stirring in a rotating manner such as using a stirring body or a shaker, the number of rotations per minute (rpm) of stirring is 10 to 3000 rpm, more preferably 50 to 1500 rpm.

In the extraction method of the present invention, the method may further include removing the extraction residue after the stirring step.

The step of removing the extraction residue may be performed using a method such as centrifugation or filtration after extraction and emulsification are sufficiently performed through the stirring.

The extraction method of the present invention as described above obtains an extract exhibiting at least one specific physiological activity among various physiological activities such as antioxidant, anti-aging, anti-inflammatory, skin whitening, skin wrinkle improvement, anti-cancer, anti-diabetic, anti-bacterial, anti-viral, etc. can be used for In particular, according to the present invention, it is useful to obtain an extract having excellent antioxidant activity from one or more extraction raw materials selected from the group consisting of mushrooms, plants and algae.

The extract extracted by the extraction method of the present invention as described above includes various useful components of biological raw materials, for example, fat-soluble substances found in cell membranes, polar substances from water-soluble parts of cells, acidic/basic ions, stable structures It can contain a complex of peroxidized structures, and these components can be stably contained in a form that exists in an environment similar to in vivo, so that excellent physiological activity can be exhibited through a network between various useful components.

Therefore, the extract extracted by the extraction method of the present invention can be used as a cosmetic or food, particularly a functional cosmetic or functional food.

The extract of the present invention may also be provided in a composition admixed with other substances. For example, it may be provided in the form of a cosmetic composition or a food composition formulated in a form mixed with cosmetically or food-acceptable substances.

At this time, there is no particular limitation on the content of the extract in the composition, for example, it may be 0.01 to 99% of the total weight of the composition.

Hereinafter, the present invention will be described in detail step by step.

ready yes. plant raw material preparation

In the present invention, the plant used for the metabolite useful component is ginseng, specifically wild ginseng cultured root (provided by Wellgreen). The ginseng (蔘) was cultured in MS medium (Murashige and Skoog medium) containing indole-3-butyric acid and sucrose, dried and crushed before use.

Example 1. Preparation of extracts and vesicles of useful plant ingredients

Comparative Example 1. Preparation of water extract of plant useful ingredients

100 ml of primary purified water was added to 5 g of ginseng of Preparation Example, and useful components were extracted while stirring at 140 rpm at room temperature for 30 minutes to 72 hours (preferably 24 hours), followed by 4000 rpm centrifugation and 0.2 um pores The filtrate obtained through a membrane filter was concentrated under reduced pressure to obtain a water extract of hemp oil.

Comparative Example 2. Preparation of ethanol extract of plant useful ingredients

After adding 100 ml of a mixture of ethanol:water = 7:3 to 5 g of the preparation example, and extracting useful components while stirring at 140 rpm at room temperature for 30 minutes to 72 hours (preferably 24 hours), centrifugation at 4000 rpm The filtrate obtained through separation and a 0.2 um pore membrane filter was concentrated under reduced pressure to obtain a water extract of hemp oil.

Comparative Example 3. Preparation of simple collection vesicles of Comparative Example 1

0.5 to 20% by weight of phospholipid (lecithin) (more specifically 2 to 6% by weight, preferably 5% by weight), 2 to 30% by weight of surfactant (more specifically PEG-8 caprylic/capric glyceride) 3 to 15% by weight, preferably 5% by weight and 0.5 to 5% by weight, preferably 1% by weight of poloxamer 407), and 0.1 to 20% by weight of lipid (more specifically, 1 to 5% by weight of cetyl ethylhexanoate) % by weight, preferably 3% by weight) was mixed and stirred at 40 to 90° C. (more specifically 60 to 90° C., preferably 80° C.) to prepare a fat-soluble part. The water-soluble part is prepared by mixing 15 to 50% by weight of a water-soluble solvent (more specifically 15 to 40% by weight of butylene glycol, preferably 30% by weight), and purified water at 20 to 90° C. (more specifically, 20 to 90° C.) It was prepared by stirring at 50 °C, preferably 30 °C). 5 g of the water-soluble part and the fat-soluble part prepared above in Comparative Example 1 were stirred while sequentially mixed and simply collected at room temperature for 30 minutes to 72 hours (preferably 24 hours). Then, Comparative Example 3 was prepared through 4000 rpm centrifugation and a 0.2 um pore membrane filter.

Comparative Example 4. Preparation of simple collection vesicles of Comparative Example 2

0.5 to 20% by weight of phospholipid (lecithin) (more specifically 2 to 6% by weight, preferably 5% by weight), 2 to 30% by weight of surfactant (more specifically PEG-8 caprylic/capric glyceride) 3 to 15% by weight, preferably 5% by weight and 0.5 to 5% by weight, preferably 1% by weight of poloxamer 407), and 0.1 to 20% by weight of lipid (more specifically, 1 to 5% by weight of cetyl ethylhexanoate) % by weight, preferably 3% by weight) was mixed and stirred at 40 to 90° C. (more specifically 60 to 90° C., preferably 80° C.) to prepare a fat-soluble part. The water-soluble part is prepared by mixing 15 to 50% by weight of a water-soluble solvent (more specifically 15 to 40% by weight of butylene glycol, preferably 30% by weight), and purified water at 20 to 90° C. (more specifically, 20 to 90° C.) It was prepared by stirring at 50 °C, preferably 30 °C). 5 g of the water-soluble part and the fat-soluble part prepared above in Comparative Example 2 were stirred while sequentially mixed and simply collected at room temperature for 30 minutes to 72 hours (preferably 24 hours). Then, Comparative Example 4 was prepared through 4000 rpm centrifugation and a 0.2 um pore membrane filter.

Comparative Example 5. Preparation of simple collection vesicles of Comparative Examples 1 and 2

0.5 to 20% by weight of phospholipid (lecithin) (more specifically 2 to 6% by weight, preferably 5% by weight), 2 to 30% by weight of surfactant (more specifically PEG-8 caprylic/capric glyceride) 3 to 15% by weight, preferably 5% by weight and 0.5 to 5% by weight, preferably 1% by weight of poloxamer 407), and 0.1 to 20% by weight of lipid (more specifically, 1 to 5% by weight of cetyl ethylhexanoate) % by weight, preferably 3% by weight) was mixed and stirred at 40 to 90° C. (more specifically 60 to 90° C., preferably 80° C.) to prepare a fat-soluble part. The water-soluble part is prepared by mixing 15 to 50% by weight of a water-soluble solvent (more specifically 15 to 40% by weight of butylene glycol, preferably 30% by weight), and purified water at 20 to 90° C. (more specifically, 20 to 90° C.) It was prepared by stirring at 50 °C, preferably 30 °C). 2.5 g of Comparative Example 1 and 2.5 g of Comparative Example 2 were mixed, stirred while sequentially mixing the water-soluble part and the fat-soluble part prepared above, and simple for 30 minutes to 72 hours (preferably 24 hours) at room temperature. captured. Then, Comparative Example 5 was prepared through 4000 rpm centrifugation and a 0.2 um pore membrane filter.

Preparation Example 1. Preparation of fusion vesicle extract of plant useful ingredients

0.5 to 20% by weight of phospholipid (lecithin) (more specifically 2 to 6% by weight, preferably 5% by weight), 2 to 30% by weight of surfactant (more specifically PEG-8 caprylic/capric glyceride) 3 to 15% by weight, preferably 5% by weight and 0.5 to 5% by weight, preferably 1% by weight of poloxamer 407), and 0.1 to 20% by weight of lipid (more specifically, 1 to 5% by weight of cetyl ethylhexanoate) % by weight, preferably 3% by weight) was mixed and stirred at 40 to 90° C. (more specifically 60 to 90° C., preferably 80° C.) to prepare a fat-soluble part. The water-soluble part is prepared by mixing 15 to 50% by weight of a water-soluble solvent (more specifically 15 to 40% by weight of butylene glycol, preferably 30% by weight), and purified water at 20 to 90° C. (more specifically, 20 to 90° C.) It was prepared by stirring at 50 °C, preferably 30 °C). After sequentially mixing and stirring the water-soluble part and the fat-soluble part prepared above to 5 g of 3 g of Preparation Example and stirring at 140 rpm at room temperature for 30 minutes to 72 hours (preferably 24 hours), the useful components are extracted, A fusion vesicle extract (Preparation Example 1) in which useful components of the metabolites of hemp was extracted and collected at the same time through 4000 rpm centrifugation and a 0.2 um pore membrane filter was prepared.

Example 2. plant Check the effect of extracts of useful ingredients and vesicles

Example 2-1. Cytotoxicity check

MTT tetrazolium is a yellow aqueous solution. When cells are treated, the metabolic process is reduced to blue-violet insoluble MTT formazan crystals as the ring structure of tetrazolium is broken by dehydrogenase in the mitochondria of intact cells. It is an experimental method mainly used to evaluate the viability of cells for a sample. Cell viability measurement according to the present invention was evaluated by measuring the absorbance of MTT formazan.

In order to confirm the cytotoxicity of Comparative Examples 1 to 5 and Preparation Example 1, human keratinocytes (HaCaT) were treated with 10% fetal bovine serum (FBS, Gibco, USA) and 1% penicillin-streptomycin (penicillin-streptomycin). , Gibco, USA) was added to DMEM medium (Dulbecco Modified Eagle Medium) and _{cultured at 37° C., 5% CO 2} conditions and dispensed at 1 ^{×10 4} cells/well in a 96-well plate. After 24 hours, the samples of Comparative Examples 1 to 5, or Preparation Example 1 were diluted in the medium and treated to a concentration of 0.25 to 4%, incubated for an additional 48 hours, and treated with an MTT solution having a concentration of 0.5 mg/ml. After 4 hours, the formed MTT formazan was dissolved in 200 μl of DMSO (dimethyl sulfoxide) and absorbance at 420 nm (SpectraMax i3, Molecular devices, CA, USA) was measured. Cell viability was calculated by Equation 1 below, and the results are shown in Table 1 below.

[Equation 1]

Cell activity (%) = (absorbance in the sample treated group / absorbance in the untreated group) * 100

As a result of the experiment, it was confirmed that Comparative Examples 1 to 5, and Preparation Example 1 did not have any significant cytotoxicity.

Example 2-2. Check the antioxidant effect

DPPH (2,2-diphenyl-1-picrylhydrazyl) is a chemically stabilized, water-soluble free radical, which is reduced by aromatic compounds and aromatic amines, and the radicals are removed, and the existing purple to yellow color is discolored. That is, the greater the reducing power, the greater the antioxidant ability is evaluated, and there is an advantage in that the effect can be confirmed in a short time by the absorbance at 517 nm. The antioxidant effect of the extract according to the present invention was evaluated by measuring the DPPH radical scavenging ability by absorbance.

To this end, each of Comparative Examples 1 to 5 or Preparation Example 1 having a 0.25 to 4% concentration of 200 μl was added to 200 μl of the 0.2 mM DPPH solution and reacted at room temperature for 20 minutes, and then absorbance at 517 nm (SpectraMax i3, Molecular devices, CA, USA) were measured. Antioxidant activity (scavenging activity) was calculated by Equation 2 below, and the results are shown in Table 2 below.

[Equation 2]

Antioxidant activity (%) = (1-absorbance in the sample treated group / absorbance in the untreated group) * 100

As a result of the experiment, it was confirmed that the antioxidant ability of Preparation Example 1 was very excellent compared to Comparative Examples 1 to 5 at all concentrations tested.

Example 2-3. Check the antibacterial effect

The minimum concentration at which a drug, such as an antibacterial agent, inhibits the growth of bacteria, usually expressed as MIC (Minimal Inhibitory Concentration). According to the properties of the medium, it is divided into agar dilution method and liquid medium dilution method. When a certain amount of strain is inoculated after diluting the test antimicrobial agent two-fold in the basal medium, the minimum inhibitory concentration at which the growth of microorganisms does not occur is called MIC. The minimum concentration is determined as MIC.

After each 50 μl of TSB (Tryptic Soy broth) and PDB (Potato Dextrose broth) was dispensed in a 96-well plate, 100 μl of each of Comparative Examples 1 to 5, or Preparation Example 1 diluted two-fold, was added, and 4 types of the test strain was inoculated. After culturing them for 1 day in a general incubator at 35±2° C., the minimum concentration of antibiotics that inhibited proliferation was determined as the MIC by visual observation, and the results are shown in Table 3.

As a result of the experiment, it was confirmed that the antibacterial efficacy of Preparation Example 1 was the best compared to Comparative Examples 1 to 5 in all the tested strains.

Example 2-4. Confirmation of the effect of inhibiting collagenase activity

In the skin composed of the epidermis, dermis, and subcutaneous tissue, collagen is a major structural protein occupying 80 to 90% of the dermal layer. On the other hand, collagenase and elastase, which are expressed in ultraviolet rays and free radicals, etc., decompose collagen and elastin to induce a decrease in skin elasticity and generation of wrinkles. It is known that by lowering the activity of collagenase that specifically acts on collagen, it is possible to maintain the mechanical properties of the skin tissue to maintain elasticity and prevent the skin from sagging. Therefore, the anti-wrinkle effect was evaluated by confirming the collagenase activity inhibitory ability of the extract according to the present invention.

_{To this end, 4 mM CaCl 2} is added to 0.1 M Tris-HCl buffer (pH7.5), and 0.3 mg/ml of 4-phenyl azobenzyloxycarbonyl-Pro-Leu-Gly-Pro-D-Arg is dissolved to prepare a substrate solution. did. Prepare the Comparative Examples 1 to 5 or Preparation Example 1 at a concentration of 0.25 to 4%, and add 75 µl of 0.2 mg/ml collagenase to a mixture of 125 µl of the substrate solution and 50 µl of the extract sample, and then at room temperature for 20 minutes. reacted. Then, 250 μl of 6% citric acid was added to stop the reaction, and 1.5 ml of ethyl acetate was added to measure absorbance at 320 nm (SpectraMax i3, Molecular devices, CA, USA). The ability to inhibit collagenase activity was calculated by Equation 3 below, and the results are shown in Table 4 below.

[Equation 3]

Collagenase activity inhibition ability (%) = (1-absorbance in the sample treated group / absorbance in the untreated group) * 100

As a result of the experiment, it was confirmed that the collagenase activity inhibitory ability of Preparation Example 1 was the best compared to Comparative Examples 1 to 5 at all concentrations tested.

Example 2-5. Pro-collagen type 1 synthesis promoting effect

Collagen is a representative fibrous protein present throughout the body, and there are 14 types, of which collagen type 1 is mainly present in the skin. When oxidative stress is applied to skin cells by external harmful factors such as ultraviolet rays, cigarette smoke, and pollutants, the production of collagen type 1 decreases, which causes wrinkles. Therefore, by evaluating the expression level of pro-collagen type 1 stimulated with _{H 2} O ₂ of the extract according to the present invention, the anti-wrinkle efficacy was confirmed.

To this end, human keratinocytes were each dispensed at 5X10 ⁴ cells/well in a 24-well plate and cultured at 37° C. and 5% CO ₂ conditions for 24 hours. Thereafter, Comparative Examples 1 to 5, or Preparation Example 1 was diluted in DMEM medium to a concentration of 2%, and the cells were pretreated for 1 hour, and then further treated with 20 μM H ₂ O _{2 and} cultured for 24 hours. After that, the amount of collagen synthesized by obtaining the supernatant of each well was evaluated using a PIP EIA kit (Procollagen Type 1 C-PeptideEnzyme Immuno Assay KIT), and the results are shown in FIG. 1 .

As a result of the experiment, it was confirmed that Preparation Example 1 promotes collagen production by protecting the collagen production inhibition induced by _{H 2} O _{2 compared to Comparative Examples 1 to 5 .}

Example 3. Preparation of a cosmetic composition comprising a fusion vesicle extract (Preparation Example 1)

The fusion vesicle extract (Preparation Example 1), in which the metabolite useful component of the present invention is extracted and collected at the same time, is well compatible with other ingredients constituting the cosmetic composition, so that cosmetics such as essence, softening lotion, nutrient emulsion, and nutrient cream are stably applied. In order to confirm whether it is made, cosmetic compositions were prepared with the compositions shown in Tables 5 to 8 below.

Table 5 shows the essence, Table 6 shows the softening lotion, Table 7 shows the nutrient emulsion, and Table 8 shows the composition of the nutrient cream.

As a result of the experiment, it was confirmed that the fusion vesicle extract of the present invention was well compatible with conventional cosmetic ingredients, so that cosmetic formulations such as essence, softening lotion, nutrient emulsion, and nutrient cream were well prepared.

As described above in detail a specific part of the present invention, for those of ordinary skill in the art, this specific description is only a preferred embodiment, and it is clear that the scope of the present invention is not limited thereto. Accordingly, the substantial scope of the present invention will be defined by the appended claims and their equivalents.

Claims (13)

(a) mixing a phospholipid, a surfactant, and a lipid to prepare a fat-soluble part;
(b) preparing a water-soluble part by mixing a water-soluble solvent and purified water; and;
(c) simultaneously adding (a) and (b) to hemp and stirring; as a method for producing a vesicle in which useful ingredients of hemp (蔘) metabolites are collected,
The method is that the extraction of metabolite useful components from hemp and the formation of the vesicles in which the extract is collected are carried out at the same time, the method.
The method of claim 1,
The surfactant in step (a) is a span-type surfactant, a tween-type surfactant, or a nonionic surfactant.
The method of claim 1,
The lipid in step (a) is a lipid having 12 or more carbon atoms, the production method.
4. The method of claim 3,
The lipid is a fatty acid ester (fatty acid ester), or a vegetable oil, the production method.
The method of claim 1,
The water-soluble solvent in step (b) is an alcohol having 2 to 5 carbon atoms, the manufacturing method.
The method of claim 1,
The metabolite useful component is a low-molecular substance present in hemp, which is a component comprising a fat-soluble substance found in the cell membrane, a polar substance from water-soluble parts of the cell, an acidic ion, and a basic ion in combination. Way.
delete A vesicle prepared by the method of claim 1, in which hemp metabolite useful ingredients are collected.
A cosmetic composition comprising the vesicle of claim 8.
10. The method of claim 9,
The cosmetic composition is skin antioxidant, wrinkle improvement, or will have an antibacterial effect, the cosmetic composition.
A food composition comprising the vesicle of claim 8.
12. The method of claim 11,
The food composition is a skin antioxidant, wrinkle improvement, or will have an antibacterial effect, the food composition. delete

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