KR20220071711A - composition containing ginseng fruit - Google Patents

Abstract

The present invention relates to a health functional food composition, a cosmetic composition, and a pharmaceutical composition containing a green berry ginseng fruit extract or an enzyme treated extract thereof as an active ingredient, and can be used for developing a whitening agent and an anti-inflammation agent.

Description

Composition containing ginseng fruit

The present invention relates to a composition comprising ginseng fruit, and more particularly, to the whitening and anti-inflammatory use of a novel greenberry ginseng fruit extract or an enzyme-treated extract thereof for skin cells.

Ginseng is a perennial semi-shady plant, belonging to the family Ogapiaceae, and its efficacy is widely known in various Chinese and Korean medical books, and has been used as a herbal medicine in many fields since ancient times. In modern times, ginseng has been widely studied, and ginseng is known to contain a large amount of ginsenosides (saponin). Ginseng usually takes more than 4 years of cultivation, and in order to obtain the highest quality ginseng, it must be cultivated for 6 years. There are disadvantages in that it is less expensive, and it takes a lot of time to obtain ginseng. Ginseng has roots and brain heads under the ground, and consists of stems, leaves, and fruits above the ground. More specifically, it is a part connecting the roots of the fine root, slow root, and main root under the ground and the brain head, the stem connected from the brain head, the leaf extending from the stem, the fruit (flower, fruit) at the end of the stem, and the part connecting the stem and the fruit. There is a vase with branches. As for ginseng, ginseng flowers start to bloom in the 3rd year or more, and green colored fruits appear around May. In mid-July, the green fruits of ginseng ripen and turn red to bear fruit. Ginsenoside means saponin contained in ginseng, and ginseng contains various types of ginsenoside components. Ginsenosides of ginseng include Panaxadiol (PD), Panaxatriol (PT), and Oleanane. In addition to the above ginsenoside components, ginseng contains carbohydrates such as starch as non-ginsenoside substances, antioxidant aromatic compounds of polyacetylene, gomisin N-A that protects the liver, and acidic peptides with insulin-like action. The main pharmacological action of ginsenoside is to exert various effects on body control functions by affecting the central nervous system, endocrine system, immune system, and metabolic system. In addition, ginsenoside (1) has a large fat-decomposing power, promotes nutrient absorption and digestion, (2) promotes metabolism by activating enzymes in cells, (3) increases energy to restore vitality, fatigue, and feeling of helplessness , anorexia improvement, (4) is known to have effects such as promoting serum protein synthesis.

The stem and leaves of ginseng affect root development through photosynthesis, but the fruit rather consumes nutrients and inhibits root growth and is a by-product that is discarded. Therefore, ginseng farms mainly use the roots of ginseng, so to promote root growth rather than reproductive growth, ginseng pyrotechnics are removed before and after the flowering period and discarded. However, recently, research results revealed that the ginsenoside content of ginseng fruit is higher than that of the root, and methods to utilize it are being studied.

Under this background, the present inventors determined that the ginseng fruit extract will be used for whitening and anti-inflammatory, and as a result of earnest efforts to solve the conventional problems, confirmed that the green berry ginseng fruit has excellent whitening and anti-inflammatory effects, thereby completing the present invention did

One object of the present invention is to provide a health functional food composition comprising a green berry ginseng fruit extract or an enzyme-treated extract thereof as an active ingredient.

Another object of the present invention is to provide a cosmetic composition comprising a green berry ginseng fruit extract or an enzyme-treated extract thereof as an active ingredient.

Another object of the present invention is to provide a pharmaceutical composition for preventing or treating inflammation-related diseases, comprising an extract of green berry ginseng fruit or an enzyme-treated extract thereof as an active ingredient.

Another object of the present invention is to provide a method for preventing or treating inflammation comprising administering the pharmaceutical composition of the present invention to a subject other than a human.

Another object of the present invention comprises a first step of harvesting greenberry ginseng fruit, a second step of extracting a ginseng fruit extract through the ginseng fruit, and a third step of enzymatically treating the ginseng fruit extract, To provide a method for preparing an enzyme-treated extract of ginseng fruit extract.

This will be described in detail as follows. Meanwhile, each description and embodiment disclosed in the present invention may be applied to each other description and embodiment. That is, all combinations of the various elements disclosed herein fall within the scope of the present invention. In addition, it cannot be considered that the scope of the present invention is limited by the specific descriptions described below.

A first aspect of the present invention for achieving the above object provides a health functional food composition comprising a green berry ginseng fruit extract or an enzyme-treated extract thereof as an active ingredient.

In addition, a second aspect of the present invention provides a cosmetic composition comprising a green berry ginseng fruit extract or an enzyme-treated extract thereof as an active ingredient.

In addition, a third aspect of the present invention provides a pharmaceutical composition for preventing or treating inflammation-related diseases, comprising the green berry ginseng fruit extract or an enzyme-treated extract thereof as an active ingredient.

In addition, a fourth aspect of the present invention provides a method for preventing or treating inflammation comprising administering the pharmaceutical composition of the present invention to an individual other than a human.

In addition, a fifth aspect of the present invention comprises a first step of harvesting greenberry ginseng fruit, a second step of extracting a ginseng fruit extract through the ginseng fruit, and a third step of enzymatically treating the ginseng fruit extract, Provided is a method for preparing an enzyme-treated extract of ginseng fruit extract.

Hereinafter, the present invention will be described in more detail.

An embodiment of the present invention for achieving the above object is that the green berry ginseng fruit extract or the enzyme-treated extract thereof according to the present invention exhibits an excellent anti-inflammatory effect, and thus a health functional food composition for the purpose of improving whitening, preventing or improving inflammation Since it is possible to consume the health functional food composition on a daily basis, a high effect can be expected for whitening improvement or prevention or improvement of inflammation.

As used herein, the term "green berry cultivar K-1, GK-1" may be denoted as K-1 or GK-1, and refers to ginseng fruit harvested from green berry ginseng.

As used herein, the term "extract" means an isolated substance obtained from a liquid or solid mixture by a solvent in a conventional manner.

The extract may be extracted with a solvent that is water, a C1 to C4 alcohol, or a mixture thereof. Specifically, the alcohol may be ethanol or methanol, but is not limited thereto, and may be extracted with a known solvent. More specifically, it may be extracted using water or distilled water as a solvent.

As the extraction method, conventional methods in the art, such as filtration, hot water extraction, immersion extraction, reflux cooling extraction, and ultrasonic extraction, may be used. In the hot water extraction method, extraction may be performed 1 to 5 times, and specifically, extraction may be repeated 3 times, but is not limited thereto.

The extraction solvent may be added 0.1 to 10 times to the dried ginseng, specifically 0.3 to 5 times, but is not limited thereto. In addition, the extraction temperature may be specifically 20 ℃ to 40 ℃, but is not limited thereto, and the extraction time may specifically be 12 to 48 hours, but is not limited thereto.

The enzyme treatment is performed through a recombinant enzyme, and the recombinant enzyme may be beta-glucosidase protein 1 (BGP1).

As used herein, the term "enzyme treatment" refers to converting ginsenosides contained in ginseng fruit into ginsenosides with high economic value without side effects.

Specifically, the enzyme treatment of the present invention may convert ginsenoside Re into ginsenoside Rg2 through hydrolysis of glucose units, and ginsenoside Rg2 exhibits stronger neuroprotective and obesity prevention properties compared to Re, Because of its high economic value as a food, drug and cosmetic material, it may be converted to recombinant enzyme treatment to provide anti-inflammatory and whitening effects.

In a specific example, as a result of comparing the greenberry ginseng fruit extract (GK-1) without enzyme treatment with the enzyme-treated greenberry ginseng fruit extract (BGK-1) of the present invention, the greenberry ginseng fruit extract of the present invention (BGK-1) suppresses NF-κB, TNF-α, iNOS, and NO production in a chain to have better anti-inflammatory effect, and suppresses melanin production and tyrosinase activity to have better whitening effect, NO production It can be confirmed that the anti-inflammatory effect is excellent, such as inhibiting.

As used herein, the term “health functional food” refers to food manufactured and processed using raw materials or ingredients useful for the human body in accordance with Act No. 6727 of the Health Functional Food Act, and 'functionality' refers to the structure of the human body. And it means to obtain a useful effect for health use, such as regulating nutrients for function or physiological action. On the other hand, health food means food that has an active health maintenance or promotion effect compared to general food, and health supplement means food for the purpose of health supplementation. In some cases, the terms health functional food, health food and health supplement food can be mixed.

The green berry ginseng fruit extract or the enzyme-treated extract thereof of the present invention may be added as it is or used together with other foods or food ingredients, and may be appropriately used according to a conventional method.

The food of the present invention can be prepared by a method commonly used in the art, and at the time of manufacture, it can be prepared by adding raw materials and components commonly added in the art. Specifically, the food composition may further include a physiologically acceptable carrier, the type of carrier is not particularly limited and any carrier commonly used in the art may be used. In addition, the food composition contains food additives such as preservatives, disinfectants, antioxidants, colorants, coloring agents, bleaching agents, seasonings, sweeteners, flavoring agents, expanding agents, strengthening agents, emulsifying agents, thickeners, filming agents, gum base agents, foam inhibitors, solvents, and improving agents. may include The additive may be selected according to the type of food and used in an appropriate amount.

In addition, the formulation of the food may be prepared without limitation as long as it is a formulation recognized as a food. The composition for food of the present invention can be prepared in various forms, and unlike general drugs, it has the advantage that there are no side effects that may occur when taking the drug for a long period of time using food as a raw material, and it is excellent in portability, so the present invention Foods of can be ingested as an adjuvant to enhance the effect of prevention or improvement of inflammation.

The green berry ginseng fruit extract or the enzyme-treated extract thereof of the present invention may be included in various weight % in the food composition if it can exhibit the effect of improving whitening, preventing or improving inflammation. Specifically, it may be included in an amount of 0.00001 to 100% by weight or 0.01 to 80% by weight relative to the total weight of the food composition, but is not limited thereto. In the case of long-term ingestion for health and hygiene purposes, it may contain an amount below the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount above the above range.

As used herein, the term “whitening” refers to improving skin tone by brightening the skin tone. Specifically, it may be to exhibit a whitening effect by inhibiting tyrosinase activity involved in melanin synthesis and melanin production.

As used herein, the term "melanin" refers to one of the pigments, which is dark brown or black, and is produced in melanocytes present in the basal layer of the epidermis. Specifically, when a lot of melanin is produced, the color of the skin may become dark.

As used herein, the term "tyrosinase (tyrosinase)" refers to an enzyme that oxidizes tyrosine by molecular oxygen to produce melanin.

In a specific embodiment of the present invention, it was confirmed that the green berry ginseng fruit extract or its enzyme-treated extract had the effect of inhibiting melanin production and tyrosinase activity, thereby having a whitening effect.

As used herein, the term “anti-inflammatory” refers to inhibiting or preventing the occurrence of inflammation, and is not particularly limited, but may be caused by cell damage or external infectious agents.

The "inflammation" refers to tissue (cell) damage or infection with external infectious agents (bacteria, viruses, fungi, various types of allergens), enzyme-activated inflammation due to the involvement of various inflammatory mediators and immune cells in local blood vessels and body fluids It exhibits a series of complex physiological reactions such as mediator secretion, body fluid infiltration, cell migration, and tissue destruction, and external symptoms such as erythema, edema, fever, and pain. In normal cases, the inflammatory response removes external infectious agents and regenerates damaged tissues to restore the function of life, but if the antigen is not removed or the internal substance is the cause and the inflammatory response is excessive or continuous, it rather promotes mucosal damage. diseases such as cancer. More specifically, inflammation may include, but is not limited to, colitis.

As used herein, the term “inflammation-related disease” refers to a disease in which inflammation is the main lesion. Inflammation is a non-specific response mediated by innate immunity, and when caused by a pathogen, it occurs regardless of the type of pathogen or previous infection. In addition to innate immunity, some local adaptive immunity are also involved in inflammation. Inflammation has a protective function to cure infection or promote tissue regeneration, but at the same time, tissue damage or disease can occur as a result of inflammation. There is also a method of intentionally inducing an inflammatory response, such as prolotherapy, to attempt tissue regeneration. Specifically, it may include colitis, Behcet's disease, Crohn's disease, rheumatoid arthritis, hair follicles, seborrheic dermatitis, psoriasis, lupus, chronic obstructive pulmonary disease, irritable bowel disease, digestive steatosis, vasculitis, and the like.

In a specific embodiment of the present invention, it was confirmed that the green berry ginseng fruit extract or its enzyme-treated extract had NO production inhibitory activity and inflammation-related cytokine (TNF-α) activity inhibitory effect.

As used herein, the term "inhibition of NO production" refers to inhibiting the promotion of inflammatory responses such as vascular permeability and edema by the generation of NO appearing in an inflammatory state. Specifically, NO not only promotes inflammatory reactions such as vascular permeability and edema, but also promotes biosynthesis of inflammatory mediators, expands blood vessels, inhibits cancer tissue growth, inhibits proliferation of viruses or bacteria, or directly kills, etc. It plays an important role in the body's immune-inflammatory response. Therefore, it is essential to examine the inhibition of NO production for substances having anti-inflammatory activity, and the anti-inflammatory effect can be confirmed by the inhibition rate of NO production.

As used herein, the term “cytokine” refers to proteins secreted by immune cells. After cytokines are secreted from cells, they can affect other cells or the cells themselves. That is, it induces the proliferation of macrophages or promotes the differentiation of secretory cells themselves. Cytokines act through receptors and are particularly important in the immune system. Cytokines are also important in maintaining health and in disease, especially in the body's response to infections, immune responses, inflammation, trauma, sepsis, cancer and reproduction.

When pathogens and viruses enter the host, it recognizes the pathogen and sends a signal to activate the transcription factor NF-kB, which induces inflammatory gene material (cytokine) to cause various diseases. It was confirmed that the green berry ginseng fruit extract of the present invention or an enzyme-treated extract thereof exhibits an anti-inflammatory effect by inhibiting NO production and cytokine (eg, TNF-α) activity.

Therefore, it was confirmed that the green berry ginseng fruit extract or the enzyme-treated extract thereof has an anti-inflammatory use through the specific example, which further suggests that it can be used for preventing or treating inflammation-related diseases.

As used herein, the term “prevention” refers to any action that inhibits or delays the onset of inflammation by administration of the composition according to the present invention.

As used herein, the term "improvement" refers to any action that at least reduces a parameter, for example, the severity of a symptom, associated with a condition to be treated by administration of a composition according to the present invention.

As used herein, the term "treatment" refers to any action in which the symptoms of the subject suspected of inflammation and the onset of inflammation are improved or beneficially changed by administration of the composition according to the present invention.

Another embodiment of the present invention for achieving the above object is that the green berry ginseng fruit extract or the enzyme-treated extract thereof according to the present invention exhibits an excellent anti-inflammatory effect, so it is added to a cosmetic composition for the purpose of improving whitening, preventing or improving inflammation. It may be included, and since the cosmetic composition can be applied routinely, a high effect can be expected for the prevention or improvement of inflammation.

In this case, the descriptions of “greenberry ginseng fruit”, “whitening”, “inflammation”, “improvement”, “treatment”, and “prevention” are the same as described above.

As used herein, the term "cosmetic composition" refers to a composition containing the compound, and the formulation may be in any form. For example, cosmetics prepared using the composition include creams such as nourishing creams, eye creams, massage creams, cleansing creams, packs, lotions such as nourishing lotions, essences, softening lotions, and lotions such as nourishing lotions. These are liquids, powders, foundations and makeup bases, and in order to achieve the object of the present invention, they may be manufactured and commercialized in any form of these formulations, and are not limited to the above examples. In addition, the cosmetic composition according to the present invention may be formulated by a conventional cosmetic preparation method. Specifically, the cosmetic of the present invention is a skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisture lotion, nourishing lotion, massage cream, nourishing cream, moisture cream, hand cream, essence, pack, mask pack, mask sheet , soap, shampoo, cleansing foam, cleansing lotion, cleansing cream, body lotion, body cleanser, emulsion, press powder, loose powder, and eye shadow may have any one formulation selected from the group consisting of. The cosmetic composition of the present invention may include an extract of green berry ginseng fruit or an enzyme-treated extract thereof or other additives such as excipients and carriers in addition thereto.

Specifically, the cosmetic composition of the present invention may further include a transdermal penetration enhancer. As used herein, the term, transdermal penetration enhancer is a composition that allows a desired component to penetrate into the blood vessel cells of the skin at a high absorption rate. Preferably, other phospholipid components used in lecithin cosmetics, liposome components, etc. are included, but are not limited thereto. In addition, as an oil that can be mainly used as an oily component, at least one selected from vegetable oil, mineral oil, silicone oil, and synthetic oil may be used. More specifically, mineral oil, cyclomethicone, squalane, octyldodecyl myristate, olive oil, Vitis vinifera seed oil, macadamia nut oil, glyceryl octanoate, castor oil, ethylhexyl isononanoate, dime Chicon, cyclopentasiloxane and sunflower seed oil can be used. In addition, 0.1 to 5% by weight of a surfactant, a higher alcohol, etc. may be added to reinforce the emulsification ability. Conventional surfactants such as nonionic surfactants, anionic surfactants, cationic surfactants, amphoteric surfactants, and phospholipids can be used as the surfactant, and specifically, sorbitan sesquinoleate, polysorbate 60 , glyceryl stearate, lipophilic glyceryl stearate, sorbitan oleate, sorbitan stearate, DA-cetyl phosphate, sorbitan stearate/sucrose cocoate, glyceryl stearate/polyethylene glycol-100 Stearate, ceteareth-6 olivate, arachidyl alcohol/behenyl alcohol/arachidyl glucoside, polypropylene glycol-26-buteth-26/polyethylene glycol-40 hydrogenated castor oil, etc. can be used. have. As the higher alcohol, an alcohol having 12 to 20 carbon atoms, such as cetyl alcohol, stearyl alcohol, octyldodecanol, isostearyl alcohol, and the like, may be used alone or in combination of one or more. The aqueous phase component may further add 0.001 to 5% by weight of one or more thickeners such as carbomer, xanthan gum, bentonite, magnesium aluminum silicate, cellulose gum, dextrin palmitate, and the like to adjust the viscosity or hardness of the aqueous phase. In addition, in the cosmetic composition of the present invention, if necessary, active ingredients such as higher fatty acids and vitamins, sunscreens, antioxidants (butylhydroxyanisole, propyl gallic acid, elisorbic acid, tocopheryl acetate, butylated hydroxy toluene, etc.), preservatives (methylparaben, butylparaben, propylparaben, phenoxyethanol, imidazolidinylurea, chlorphenesin, etc.), colorants, pH adjusters (triethanolamine, citric acid, citric acid, sodium citrate, malic acid, Sodium malate, fmalic acid, sodium fumarate, succinic acid, sodium succinate, sodium hydroxide, sodium monohydrogen phosphate, etc.), humectants (glycerin, sorbitol, propylene glycol, butylene glycol, hexylene glycol, diglycerin) , betaine, glycereth-26, methylgluceth-20, etc.), lubricants, etc. may be further added. In addition, the cosmetic composition of the present invention additionally includes a material that can supplementally provide essential nutrients to the skin, and preferably contains an auxiliary agent including, but not limited to, natural fragrance, cosmetic fragrance, or herbal medicine. have.

Another embodiment of the present invention for achieving the above object is characterized in that the pharmaceutical composition comprising the green berry ginseng fruit extract or an enzyme-treated extract thereof according to the present invention as an active ingredient is for preventing or treating inflammation-related diseases.

In this case, the descriptions of “greenberry ginseng fruit”, “inflammatory-related disease”, “inflammation”, “improvement”, “treatment”, and “prevention” are the same as described above.

The pharmaceutical composition of the present invention may further include a pharmaceutically acceptable salt.

As used herein, the term "pharmaceutically acceptable salt" refers to a salt in a form that can be used pharmaceutically among salts, which are substances in which a cation and an anion are combined by electrostatic attraction, and is usually a metal salt, an organic base and salts, salts with inorganic acids, salts with organic acids, salts with basic or acidic amino acids, and the like. For example, the metal salt may be an alkali metal salt (sodium salt, potassium salt, etc.), alkaline earth metal salt (calcium salt, magnesium salt, barium salt, etc.), an aluminum salt or the like; Salts with organic bases include triethylamine, pyridine, picoline, 2,6-lutidine, ethanolamine, diethanolamine, triethanolamine, cyclohexylamine, dicyclohexylamine, N,N-dibenzylethylenediamine salts with, etc.; salts with inorganic acids may be salts with hydrochloric acid, hydrobromic acid, nitric acid, sulfuric acid, phosphoric acid and the like; Salts with organic acids may be salts with formic acid, acetic acid, trifluoroacetic acid, phthalic acid, fumaric acid, oxalic acid, tartaric acid, maleic acid, citric acid, succinic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, and the like; Salts with basic amino acids may be salts with arginine, lysine, ornithine and the like; The salt with an acidic amino acid may be a salt with aspartic acid, glutamic acid, or the like.

The pharmaceutical composition of the present invention may further include a pharmaceutically acceptable carrier, excipient or diluent, wherein the carrier may include a non-naturallyoccuring carrier.

Specifically, carriers, excipients and diluents that may be included in the pharmaceutical composition include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium Silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, polycaprolactone, polylactic acid Lactic Acid), poly-L-lactic acid, and mineral oil. The pharmaceutical composition may be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, etc., external preparations, suppositories, and sterile injection solutions, respectively, according to conventional methods. The form may include various amorphous carriers, microspheres, nanofibers, and the like. In the case of formulation, it can be prepared using a diluent or excipient such as a filler, extender, binder, wetting agent, disintegrant, surfactant, etc. commonly used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and these solid preparations include at least one excipient in the extract and its fractions, for example, starch, calcium carbonate, It may be prepared by mixing sucrose or lactose, gelatin, or the like. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used. Liquid formulations for oral administration include suspensions, solutions, emulsions, syrups, etc. In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included. have. Formulations for parenteral administration may include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, suppositories, and the like. Non-aqueous solvents and suspending agents include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. The content of the green berry ginseng fruit extract or the enzyme-treated extract or pharmaceutically acceptable salt thereof contained in the pharmaceutical composition of the present invention is not particularly limited. Another embodiment of the present invention for achieving the above object provides a health functional food composition for preventing or improving inflammation comprising the green berry ginseng fruit extract or an enzyme-treated extract thereof.

Another aspect of the present invention for achieving the above object is a first step of harvesting greenberry ginseng fruit, a second step of extracting a ginseng fruit extract through the ginseng fruit, and a second step of enzymatically treating the ginseng fruit extract It provides a method for preparing an enzyme-treated extract of ginseng fruit extract, comprising three steps.

Specifically, the green berry ginseng fruit extract prepared by the above manufacturing method or its enzyme-treated extract converts ginsenoside into ginsenoside with high economic value as a food, drug and cosmetic material without side effects compared to the conventional method for preparing an extract. effect can be provided.

The harvest may be to harvest the ginseng fruit 14 to 21 days after the flowering period. If the harvest is less than 14 days or more than 21 days from the flowering period, the total ginsenoside content contained in the green berry ginseng fruit is low, so the whitening and anti-inflammatory effects may be insufficient.

The enzyme may be a beta-glucosidase protein 1 (BGP1) recombinant enzyme.

As used herein, the term "enzyme treatment" refers to converting ginsenosides contained in ginseng fruit into ginsenosides with high economic value without side effects.

Specifically, the enzyme treatment of the present invention may convert ginsenoside Re into ginsenoside Rg2 through hydrolysis of glucose units, and ginsenoside Rg2 exhibits stronger neuroprotective and obesity prevention properties compared to Re, Because of its high economic value as a food, drug and cosmetic material, it may be converted to recombinant enzyme treatment to provide anti-inflammatory and whitening effects. In a specific example, as a result of comparing the greenberry ginseng fruit extract (GK-1) without enzyme treatment with the enzyme-treated greenberry ginseng fruit extract (BGK-1) of the present invention, the greenberry ginseng fruit extract of the present invention It can be seen that (BGK-1) suppresses NF-κB, TNF-α, iNOS, and NO production in a chain to have more excellent anti-inflammatory effect, and suppresses melanin production and tyrosinase activity to show more excellent whitening effect. .

As used herein, the term “ginsenoside” refers to steroid saponins and triterpene saponins specifically present in ginseng, and ginsenoside refers to a major active ingredient that exhibits the pharmacological effect of ginseng.

Another aspect of the present invention for achieving the above object provides a composition for preventing or improving inflammation comprising the extract prepared by the method for preparing the ginseng fruit extract as an active ingredient.

In a specific embodiment, it was confirmed that the NF-κB, TNF-α, iNOS, NO production and melanin production and tyrosinase activity of the green berry ginseng fruit extract of the present invention or the enzyme-treated extract thereof of the present invention were inhibited in a chain. did.

Therefore, it was confirmed that the green berry ginseng fruit extract of the present invention or the enzyme-treated extract of the present invention has anti-inflammatory and whitening effects through the above specific example, which can be further used as a health functional food composition, cosmetic composition, and pharmaceutical composition suggests

According to the present invention, it is confirmed that the composition comprising the green berry ginseng fruit extract of the present invention provides whitening and anti-inflammatory effects, which can be applied as a health functional food or cosmetic with high economic value by utilizing the previously discarded ginseng fruit. did

1, A is a photograph according to the growth time of greenberry ginseng fruit (K-1) ginseng fruit of the present invention, B is a graph showing the fruit weight according to the growth time, C is a ginseng root weight increase ratio according to the growth time The graph D shown is a graph showing the ginsenoside content according to the growth time.
2E is a reaction scheme showing a biological change in which ginsenoside Re is changed to Rg2 by a recombinant enzyme, F is a structural formula showing a recombinant enzyme, G is a photograph of analyzing the ginsenoside content by TLC, H is a 4-year-old green HPLC graph showing the change of ginsenoside after recombinase treatment (BGK-1) of berry ginseng fruit (GK-1), and I is after recombinase treatment of 5-year-old green ginseng fruit (GK-1) (BGK- 1) It is an HPLC graph showing the change of ginsenoside.
3 is a graph showing the whitening effect of greenberry ginseng fruit (GK-1) and enzyme-treated greenberry ginseng fruit (BGK-1) of the present invention.
4B is a graph showing the melanin content of greenberry ginseng fruit (GK-1), enzyme-treated greenberry ginseng fruit (BGK-1), arbutin, and α-MSH of the present invention, C is green of the present invention A graph showing the tyrosinase activity of berry ginseng fruit (GK-1), enzyme-treated green berry ginseng fruit (BGK-1), arbutin, and α-MSH.
5A is a graph showing the cytotoxicity of greenberry ginseng fruit (GK-1) and enzyme-treated greenberry ginseng fruit (BGK-1) of the present invention, B is a graph showing NO inhibition, C is iNOS gene expression Graph showing , D is a graph showing ROS generation analysis.
Figure 6E is a photograph showing the detection of oxidative stress through immunofluorescence staining of greenberry ginseng fruit (GK-1) and enzyme-treated greenberry ginseng fruit (BGK-1) of the present invention, F is a photograph showing detection of peroxide to be.
7G is a graph showing ROS and Mito-SOX fluorescence intensity of greenberry ginseng fruit (GK-1) and enzyme-treated greenberry ginseng fruit (BGK-1) of the present invention.
FIG. 8A is a photograph showing immunofluorescence analysis of nuclear translocation of greenberry ginseng fruit (GK-1) and enzyme-treated greenberry ginseng fruit (BGK-1) of the present invention, C is IκBα, p-IκBα, NF A photograph showing the Western blot analysis for the expression of -κB and p-NF-κB, D is a graph showing the reactive protein level of p-IκBα and IκBα.
FIG. 9B is a graph showing the NF-kB fluorescence intensity of greenberry ginseng fruit (GK-1) and enzyme-treated greenberry ginseng fruit (BGK-1) of the present invention, and E is p-NF-κB/NF- A graph showing the reactive protein level of кB, F is a graph showing NF-κB gene expression, G is a graph showing TNF-α gene expression, H is a graph showing IL-1β gene expression, I is IL-6 gene expression This is the graph shown.
Figure 10 shows the physicochemical properties of reducing sugars (A), flavonoids (B), phenols (C), acidic polysaccharides (D), and the present invention during 9 weeks of ripening of 4-year-old green berry ginseng fruit (K-1) of the present invention. A graph showing reducing sugars (E), flavonoids (F), phenols (G), and acidic polysaccharides (H) for 9 weeks of ripening of 5-year-old green berry ginseng fruit (K-1).

Hereinafter, the present invention will be described in more detail through examples. These Examples are for explaining the present invention in more detail, and the scope of the present invention is not limited by these Examples.

Example 1: Preparation of greenberry ginseng fruit extract and enzyme-treated extract thereof

Greenberry ginseng fruits harvested 2 to 4 weeks after flowering of 4 to 5 year old ginseng were provided by Gyeonggi Agricultural Research and Extension Services, Republic of Korea. Green berry cultivar K-1, GK-1 Ginseng fruit was washed, dried, pulverized, extracted with 70% ethanol, and treated with recombinant enzyme (betaglucosidase, bgp-1). An enzyme-treated extract (BGK-1) of the extract and green berry ginseng fruit extract was prepared.

As shown in FIG. 1, the green berry ginseng fruit (K-1) of the present invention is harvested from 4 to 5 year old ginseng and harvested 1 to 9 weeks after flowering, and its ginsenoside content, root weight, As a result of comparing the weight of the fruit, it was confirmed that the total ginsenoside content in the unripe and fruit harvested at 2 to 3 weeks, especially Re, was the most included.

In addition, as shown in FIG. 2 , when the recombinant enzyme was treated, it was found that the ginsenoside Re was converted into the ginsenoside Rg2 through hydrolysis of the glucose unit without side effects. It was confirmed that ginsenoside Rg2 exhibits stronger neuroprotective and anti-obesity properties compared to Re, and has high economic value as a food, drug, and cosmetic material, so that it can provide superior anti-inflammatory and whitening effects by treatment with recombinant enzymes.

Therefore, the enzyme-treated extract of the green berry ginseng fruit extract treated with the recombinant enzyme of the green berry ginseng fruit (K-1) extract of the present invention has a high content of ginsenoside Rg2, and it is used as a health functional food with high economic value or It can be seen that it can be applied as a cosmetic.

Example 2: HPLC-QTOF / MS and TLC analysis

Various ginsenoside profiling in greenberry ginseng fruit was performed via mass spectrometry (QTOF/MS) approach. This assay was run in negative ion mode on a Waters Xevo G2-S QTOF MS (Waters Corp., Milford, MA, USA).

Ginsenoside content was confirmed by HPLC (Agilent 1260, Palo Alto, CA, USA). Specifically, ginseng fruit powder was extracted with 70% ethanol, evaporated at 40° C., and dissolved in 1 ml of methanol. Thereafter, ginsenoside analysis was analyzed by HPLC on a 203 nm C18 column at a flow rate of 1.0 ml/min.

TLC evaluation was confirmed using silica gel 60 F254 plates from Merck KGaA (Darmstadt, Germany) as a developer in a chloroform-methanol-water solvent mixture with a volume ratio of 65:35:10 (v / v / v) as a developer. The spots on the TLC plate were stained by spraying sulfuric acid into ethanol (10:90, v/v) and reacting at 110° C. for 10 minutes.

Example 3: Phytochemical analysis

The reduction in the sugar content of the green berry ginseng fruit of the present invention was measured using a dinitrosalicylic acid (DNS) reagent approach. The extract prepared in Example 1 and DNS reagent were mixed, boiled for 5 minutes, and then cooled. The absorbance of the mixture was measured at 550 nm with a UV-reader (Bio-Tek, Winooski, VT) based on glucose.

The total phenol content of K-1 extract was evaluated by Folin-Ciocalteu approach. The K-1 extract dissolved in 1 ml of methanol was mixed with 10% sodium carbonate for 3 minutes. Then, 1 ml of Folin-Ciocalteu reagent was incubated for 30 minutes. The total phenol content of the ginseng fruit samples was measured using a UV reader at 750 nm and determined based on a calibration curve based on gallic acid.

As a result, as shown in FIG. 10 , it was confirmed that the total amount of ginsenosides, reducing sugars and acidic polysaccharides were the most when the five-year-old green berry ginseng fruit was harvested in the second week.

Example 4: Cell culture and viability assay

B16 and RAW 264.7 cells were cultured in Dulbecco Modified Eagle Medium (DMEM) supplemented with 10% FBS and 1% penicillin-streptomycin (100x) at 37° C., 95% humidified air, and 5% CO ₂ atmosphere. cultured in

Cell proliferation was confirmed by the MTT approach. After the cells were treated for 24 hours, 100 μL of 0.5 mg/ml MTT reagent was added and incubated at 37° C. for an additional 3 hours. Next, 100 μL of DMSO was added and absorbance was measured at 595 nm using a SpectraMax® ABS Plus microplate reader (Molecular Devices, San Jose, California, USA).

Example 5: Cell melanin content and tyrosinase activity analysis

B16 cells were treated with α-MSH (100 nM), GK-1, and BGK-1 (25 and 50 μg/mL) extracts for 72 hours, and then the melanin concentration of B16 cells was evaluated. Cells were collected and the pellet dissolved in 500 μL of NaOH with 10% DMSO at 80° C. for 1 hour. Then, the absorbance was checked at 475 nm using a microplate reader.

Meanwhile, the activity of tyrosinase, which inhibits the rate of melanin synthesis, was measured using the L-DOPA oxidase approach. After treating B16 cells, cells were lysed with PBS containing 1% Triton X-100 and freeze-thawed at -80°C for 15 minutes. After centrifugation at 120,000 rpm for 15 minutes, 10 μL of 15 mM L-DOPA was added to 90 μL of the lysate supernatant and incubated at 37° C. for 1 hour. Thereafter, the tyrosinase activity was checked for absorbance at 475 nm with a microplate reader.

As a result, as shown in FIG. 3 , a greenberry ginseng fruit extract (GK-1) without enzyme treatment on B16 cells, and an enzyme-treated extract (BGK When -1) was evaluated for cytotoxicity with MTT, it was confirmed that there was no toxic effect at all treatment concentrations of the extract.

In addition, as shown in FIG. 4, it was confirmed that melanin production was decreased in a dose-dependent manner of the extract, and it was confirmed that tyrosinase activity was also effectively inhibited. I was able to confirm that it worked. Among them, it was found that the enzyme-treated BGK-1 extract had better effects than the enzyme-treated GK-1 extract.

Example 6: Anti-inflammatory activity assay

The overproduction of NO is an inflammatory pathogenesis that reacts with ROS to cause cell damage. The production of this NO was confirmed by measuring nitrite levels using Griess reagent. Specifically, RAW.264.7 cells were pretreated with various concentrations of GK-1 extract, BGK-1 extract (25-100 μg / mL), or 1 μM dexamethasone (DEX). After 1 h, cells were stimulated with 1 μg/mL of LPS, and after 24 h incubation, the supernatant was mixed with an equal volume of Griess reagent. After this, the NO level was measured using a microplate reader at 520 nm and the sodium nitrite concentration was calculated with reference to the standard curve.

Meanwhile, the intracellular accumulation of ROS was measured by performing culture in a black 96-well plate. After treatment with RAW 264.7 cells, 10 μm 5-chloromethyl-2,7-dichlorodihydrofluorescein diacetate was incubated for 30 minutes in the dark. The emission wavelengths were 485 nm and 535 nm, respectively. To further confirm intracellular ROS generation, a cellular ROS detection assay kit was applied. Raw264.7 cells treated with GK-1 and BGK-1 for 24 hours 0.4 μL/ml oxidative stress detection and peroxide detection reagents were added to the cells. After placement for 30 min, fluorescence was measured with a laser scanning microscope (Leica, Wetzlar, Germany) and quantified using Image J software.

As a result, as shown in FIG. 5 , the greenberry ginseng fruit extract (GK-1) without enzyme treatment and the greenberry ginseng fruit enzyme extract (BGK-1) prepared by recombinantly treating the greenberry ginseng fruit were It did not show a significant cytotoxic effect on RAW 264.7 cells. In addition, iNOS induced by lipopolysaccharide (LPS) expression catalyzes NO production, and excessive NO production intensifies inflammatory response to damage tissues. -1 confirmed that both iNOS and NO production were dose-dependently inhibited, confirming that there was an anti-inflammatory effect.

In addition, as shown in FIGS. 6 to 7 , the green berry ginseng fruit extract (GK-1) and the green berry ginseng fruit extract (BGK-1) prepared by recombinantly treating the green berry ginseng fruit of the present invention were used as LPS. It was confirmed that the inhibition of the induced ROS generation. ROS is an important regulator of intracellular signal transduction and homeostasis together with molecules that chemically react with oxygen, and ROS accumulates during exposure to LPS, resulting in lethal cellular damage. In particular, BGK-1 was excellent in the effect of reducing ROS production. In conclusion, it was confirmed that GK-1 and BGK-1 could inhibit the accumulation of LPS-induced ROS in macrophages and thus have anti-inflammatory effects.

In addition, as shown in FIGS. 8 to 9 , the unenzymatically treated greenberry ginseng fruit extract (GK-1) and the greenberry ginseng fruit extract (BGK-1) prepared by recombinantly treating the greenberry ginseng fruit of the present invention were NF- It was confirmed that the activation of kB was inhibited. In RAW264.7 cells stimulated with LPS, an inflammatory response was induced and NF-κB, known as a transcription factor of the inflammatory response, was activated. to exacerbate inflammation. It was confirmed that both GK-1 and BGK-1 of the present invention sequentially inhibited NF-κB, TNF-α, iNOS, and NO production in a concentration-dependent manner. In particular, it was confirmed that BGK-1 was more effective in inhibiting inflammation compared to GK-1.

Combining Examples 1 to 6 of the present invention, the green berry ginseng fruit extract (GK-1) of the present invention and the green berry ginseng fruit enzyme-treated extract (BGK-1) prepared by recombinantly treating the green berry ginseng fruit ), both have whitening and anti-inflammatory effects. In particular, compared to the extract (GK-1) without recombinant enzyme treatment of green berry ginseng fruit, the BGK-1 extract more effectively inhibits NO production, inhibits melanin pigment, and tyrosina It was confirmed that by inhibiting the anti-inflammatory agent activity, it was possible to provide a more effective anti-inflammatory and whitening composition.

From the above description, those skilled in the art to which the present invention pertains will understand that the present invention may be embodied in other specific forms without changing the technical spirit or essential characteristics thereof. In this regard, it should be understood that the embodiments described above are illustrative in all respects and not restrictive. The scope of the present invention should be construed as being included in the scope of the present invention, rather than the above detailed description, all changes or modifications derived from the meaning and scope of the claims to be described later and their equivalents.

Claims (18)

A health functional food composition comprising a green berry ginseng fruit extract or an enzyme-treated extract thereof as an active ingredient.
According to claim 1,
The health functional food composition is for improving whitening; Or for preventing or improving inflammation, a health functional food composition.
According to claim 1,
The enzyme is beta-glucosidase protein 1 (beta-glucosidase protein 1, BGP1), the health functional food composition.
According to claim 1,
The inflammation is caused by damage to cells or external infectious agents, health functional food composition.
According to claim 1,
The inflammation is colitis, the health functional food composition.
A cosmetic composition comprising a green berry ginseng fruit extract or an enzyme-treated extract thereof as an active ingredient.
7. The method of claim 6,
The cosmetic composition is for improving whitening; Or for preventing or improving inflammation, a cosmetic composition.
7. The method of claim 6,
The enzyme is beta-glucosidase protein 1 (beta-glucosidase protein 1, BGP1), the cosmetic composition.
7. The method of claim 6,
The inflammation is caused by damage to cells or external infectious agents, the cosmetic composition.
7. The method of claim 6,
The inflammation is colitis, the cosmetic composition.
A pharmaceutical composition for preventing or treating inflammation-related diseases, comprising an extract of green berry ginseng fruit or an enzyme-treated extract thereof as an active ingredient.
12. The method of claim 11,
The enzyme is beta-glucosidase protein 1 (beta-glucosidase protein 1, BGP1), the pharmaceutical composition for preventing or treating inflammation-related diseases.
12. The method of claim 11,
The inflammation is caused by damage to cells or external infectious agents, a pharmaceutical composition for the prevention or treatment of inflammation-related diseases.
12. The method of claim 11,
The inflammation is colitis, a pharmaceutical composition for the prevention or treatment of inflammation-related diseases.
12. A method for preventing or treating inflammation comprising administering the pharmaceutical composition of claim 11 to a subject other than a human.
The first step of harvesting greenberry ginseng fruit;
a second step of extracting a ginseng fruit extract through the ginseng fruit; and
A third step of enzymatically treating the ginseng fruit extract; including, a method for producing an enzyme-treated extract of the ginseng fruit extract.
17. The method of claim 16,
The harvest is to harvest the ginseng fruit 14 to 21 days after the flowering period, the manufacturing method.
17. The method of claim 16,
The enzyme is a beta-glucosidase protein 1 (beta-glucosidase protein 1, BGP1) will be a recombinant enzyme, the manufacturing method.

Images