KR20110112915A - Composition comprising defatted green tea seed extract for preventing and treating inflammatory or cancer disease - Google Patents

Abstract

The present invention relates to an anti-inflammatory or anti-cancer composition comprising the degreased green tea seed extract as an active ingredient, and more particularly, the present invention provides an extract obtained by using water or an organic solvent for green tea seed degreased using an organic solvent for degreasing. It relates to an anti-inflammatory or anti-cancer composition containing as an active ingredient. The degreased green tea seed extract according to the present invention is excellent in antioxidant activity, and has an excellent effect of inhibiting the expression of inflammatory factors PEG2 and TNF-a, which is characterized by preventing or treating inflammatory diseases, and at the same time, The present invention has the effect of inhibiting cell proliferation and increasing the activity of detoxifying enzymes involved in the metabolism of carcinogens, as well as causing cell death of cancer cells and inducing cell death, thereby preventing or treating cancer. The composition according to the present invention has an effect that can be usefully used in the manufacture of drugs and health functional foods that can prevent and treat inflammatory diseases or cancer.

Description

Composition comprising defatted green tea seed extract for preventing and treating inflammatory or cancer disease}

The present invention relates to an anti-inflammatory or anti-cancer composition comprising a defatted green tea seed extract as an active ingredient, the anti-inflammatory or anti-cancer composition containing water or an organic solvent extract of green tea seed degreased using an organic solvent for degreasing as an active ingredient. It is about.

Inflammation is one of the defenses of biological tissues against external stimuli. When an inflammatory reaction occurs, a number of pro-inflammatory mediators are produced, resulting in clinical signs of redness, fever, swelling, pain, Symptoms such as dysfunction appear. In particular, harmful stimuli such as LPS (lipopolysaccharide), inflammation-inducing factors, and irradiation can excessively stimulate the human immune system, leading to tumor necrosis factor-α (TNF-α) and interleukin-IL-1 in immune cells such as macrophages. 1) excessively induces inflammatory mediators such as IL-6, prostagladin, luecotriene and nitric oxide (NO), resulting in inflammatory diseases, tissue transplant rejection, autoimmune diseases, It is known to cause immune diseases such as diabetes and death of nerve cells.

NO, one of the inflammatory mediators, is produced in endothelial cells or macrophages in a normal state and exhibits various physiological activities in addition to cell killing and bactericidal action. In particular, it maintains homeostasis of blood pressure in the relaxation of vascular endothelial cells. It is known to play a role. Stimulation, such as LPS, proinflammatory factors and irradiation, in particular, induces the expression of an inducible nitric oxide synthase (iNOS), which produces a large amount of NO continuously for 4-6 hours. Excessive amounts of NO may lead to such diseases. Therefore, it can be said that the development of an iNOS activity inhibitor as a therapeutic agent for inflammatory diseases is likely to be developed as a therapeutic agent for the disease.

Prostaglandins, such as PGE2, PGF2a and PGI2, are other hormones derived from arachidonic acid and are involved in various physiological activities and are regulated by cyclooxygenase (COX). Nonsteroidal anti-inflammatory drugs, such as aspirin indomethacin, which inhibit the synthesis of prostaglandins by inhibiting COX activity, are known to have excellent effects on arthritis.

In addition, TNF-α is produced in macrophages and fibroblasts activated by BCG, LPS, and inflammatory factors to induce apoptosis, production of prostaglandins and cytokines such as IL-1 and IL-6. . TNF-α found as a tumor necrosis factor has been reported to be related to autoimmune diseases such as rheumatoid arthritis and organ transplant rejection, allergic inflammatory diseases such as asthma and atopic dermatitis, and acute immune diseases such as sepsis and acute liver disease. As a result, research into compounds that inhibit the synthesis of TNF-α is being actively conducted. In addition, recent reports have shown that mice with damaged TNF-α receptors are resistant to lethal toxicity by LPS, and that mice administered with TNF-α antibodies have the effect of preventing arthritis or LPS toxicity. (Beutler B., Science, 229: 869, 1985; Pfeffer K et al., Cell , 73; 457, 1993).

On the other hand, conventional anti-inflammatory drugs that can treat these inflammatory diseases are largely divided into steroidal and non-steroidal anti-inflammatory drugs, most of the synthetic anti-inflammatory drugs may be accompanied by trilogy in the body, the effect of inflammation treatment is excellent and side effects There is a need for the development of new anti-inflammatory agents derived from natural extracts.

Tumors are collections of cells that exhibit autonomous overgrowth and are distinguished from malignant tumors that are likely to lead to death and benign tumors that do not. In particular, cancer is a genetic disease caused by mutations in genes such as oncogenes and tumor suppressor genes, which are caused at the cellular level. Oncogene products play a role in regulating the signaling system of cell division or differentiation by forming networks in cells. In other words, if their regulation is abnormal, the cell differentiation process is interrupted and indefinite division continues. If the signal transduction pathway can be reduced normally, an excellent therapeutic effect with fewer side effects can be obtained. It is expected to be.

To date, the main treatments for cancer include surgical treatment, drug therapy, and radiation therapy. In reality, when the treatment is performed alone, the therapeutic effect is insufficient. Therapies are widely practiced. However, these therapies also have the disadvantage of high cost of treatment and high side effects on the body.

Therefore, there is a need for the development of new anticancer drugs that can use pharmaceutical ingredients at low cost without side effects.

On the other hand, the recent trend of attention to the health of green tea has become more prominent as the consumption of green tea is growing in demand, the demand for green tea (Camellia sinensis L.) cultivation of green tea seeds is increasing rapidly.

In particular, green tea seeds have several pharmacological effects on folk remedies or herbal treatments, such as asthma and brain injury, chronic gastrointestinal drugs, postpartum recovery, and rapid wound healing after surgery. (Na Hyo-hwan, Baek Sun-ok, Han Sang-bin, Bok Jin-young. 1992. General Components of Green Tea Seeds J. Korean Agric Chem Soc , 35 (4): 272-275) have. In addition, green tea seeds are rich in various physiologically active ingredients such as saponins and phenols, and efforts are being made to use them for various medical purposes (Romos S. 2007. Effects of dietary flavonoids on apoptotic pathways related to cancer chemoprevention.J Nutr Biochem 18:... 427-442, Russo GL 2007. Ins and outs of dietary phytochemicals in cancer chemoprevention Biochem Pharm 74: 533-544., Vissers MN, Zock PL, Katan MB. 2004. Bioavailability and antioxidant effects of olive oil phenols in humans: a review. Eur J Clin Nutr 58 955-965).

However, there has been no report on the fraction extract of green tea seeds which can maximize the anti-inflammatory and anti-cancer activity in extracting the green tea seeds in order to further increase the industrial usefulness of green tea seeds.

Accordingly, the present inventors completed the present invention by confirming that the solvent fraction extract obtained from the green tea seed, which had been removed from the green tea seed, had an excellent anti-inflammatory and anti-cancer activity in the process of obtaining the extract containing the bioactive component from the green tea seed.

Accordingly, an object of the present invention is to provide an anti-inflammatory or anti-cancer composition comprising as an active ingredient skim green tea seed extract that can effectively prevent or treat inflammatory diseases or cancer diseases.

In addition, another object of the present invention to provide a health functional food for the prevention or improvement of inflammatory diseases or cancer diseases containing the skim green tea seed extract according to the present invention as an active ingredient.

In order to achieve the above object, the present invention provides an anti-inflammatory or anti-cancer composition comprising skim green tea seed extract as an active ingredient.

In one embodiment of the present invention, the degreasing may be to degrease at room temperature for 22 to 26 hours by adding 1500 ~ 2500ml n-hexane based on 100g green tea seed powder.

In one embodiment of the present invention, the extract may be a solvent extract solubilized by adding water or an organic solvent to the degreased green tea seed powder.

In one embodiment of the present invention, the organic solvent may be selected from the group consisting of methanol, ethanol, propanol, isopropanol, butanol, acetone, ether, benzene, chloroform, ethyl acetate and methylene chloride.

In one embodiment of the present invention, the anti-inflammation can prevent or treat inflammatory diseases selected from the group consisting of gastritis, colitis, arthritis, nephritis and hepatitis.

In one embodiment of the present invention, the anticancer may prevent or treat a cancer disease selected from the group consisting of liver cancer, stomach cancer, colon cancer, lung cancer, breast cancer, rectal cancer and pancreatic cancer.

In one embodiment of the present invention, the extract may comprise 0.1 to 50% by weight based on the total weight of the composition.

The present invention also provides a health functional food for the prevention or improvement of inflammatory diseases or cancer diseases containing the skim green tea seed extract according to the present invention as an active ingredient.

In one embodiment of the invention, the food may be in the form of powder, granules, tablets, capsules or beverages.

The degreased green tea seed extract according to the present invention is excellent in antioxidant activity, and has an excellent effect of inhibiting the expression of inflammatory factors PEG2 and TNF-a, which is characterized by preventing or treating inflammatory diseases, and at the same time, The present invention has the effect of inhibiting cell proliferation and increasing the activity of detoxifying enzymes involved in the metabolism of carcinogens, as well as causing cell death of cancer cells and inducing cell death, thereby preventing or treating cancer. The composition according to the present invention has an effect that can be usefully used in the manufacture of drugs and health functional foods that can prevent and treat inflammatory diseases or cancer.

Figure 1 shows a schematic diagram of a process for obtaining a fraction extract for each solvent of the skim green tea seed according to the present invention.
Figure 2 shows a graph measuring the cell viability after treatment of each solvent fraction extract of degreased green tea seed according to the present invention to HepG2 cells, liver cancer cell line.
Figure 3 is a graph showing the results of measuring the QR (quinone reductase) activity after treating the fractions of each solvent of the degreased green tea seed according to the present invention to HepG2 cells, liver cancer cell lines by concentration.
Figure 4 shows the result of performing Western blot expression of CYP1A1 and CYP1A2 expressed in cells after treating the fraction extract of each solvent of degreased green tea seed according to the present invention to HepG2 cells, liver cancer cell lines by concentration. Each is shown in the graph.
5 is a petroleum ether extract fraction of degreased green tea seed according to the present invention after treatment to HepG2 cells, which are liver cancer cell lines, by concentration, and the expression level of HO-1 expressed in cells through Western blotting. Respectively.
6 is a graph showing the results obtained by luciferase analysis after ethanol extract fraction, petroleum ether extract fraction, and ethyl acetate extract fraction of degreased green tea seed according to the present invention in HepG2 cells by concentration in ARE activity. will be.
7 is a graph showing the results of analyzing the effect on the cell cycle after treating ethyl acetate extract fraction (7a) and hot water extract (7b) of degreased green tea seed by concentration in HepG2 cells using a flow cytometer.
FIG. 8 shows the amount of expression of TNF-a, an inflammatory transcription factor expressed in cells after treatment with fractions of each solvent of degreased green tea seed according to the present invention, at different concentrations in HepG2 cells, liver cancer cell lines, by Western blot. The resulting photographs and graphs are shown.

The present invention is characterized by providing an anti-inflammatory or anti-cancer composition containing skim green tea seed extract as an active ingredient.

Green tea is one of foods that have been of interest in recent years as it is known to have excellent antioxidant properties among natural functional substances. Green tea contains a large number of ingredients including catechin, a polyphenolic compound, and is known to be effective in removing free radicals caused by stress in and out of the body due to its excellent pharmacological action and particularly excellent antioxidant properties (Kim MJ , et al, Nutrition Research, 22, pp733-744, 2002; Rhee SJ, et al, Kor. J. Gerontol., 8, pp 49-55, 1998).

In addition, the study of a study on the antioxidant activity of green tea Nakayama (Nakayama) (T. Nakayama, University of Shizuoka, Japan, 7, pp1991-1993, 1994) polyphenol structure of catechins are (hydrogen peroxide from the hydrogen peroxide, H ₂ O ₂ ) has been shown to have an effect of inhibiting cytotoxicity, and it has been reported that flavonoids are effective for removing single species oxygen and free radicals in the early stages of lipid peroxidation. It has been known to have excellent physiological activities such as growth inhibition, caries suppression, and cholesterol suppression of immunodeficiency virus.

On the other hand, although the green tea has excellent pharmacological efficacy and palatability, a variety of products have not been developed except for direct drying of green tea leaves in a container or tea bag, and especially in the case of green tea seeds, it is beneficial to the human body. Most of them are not commercialized even though they contain a large amount of them, and recently, it has been found that green tea seed oil, which is an oil component of green tea seeds, has a bioactive component, and an attempt to apply it to food is increasing.

Thus, in the present invention, in order to obtain an extract having very good anticancer and antiinflammatory activity from the green tea seed portion in the composition of the green tea plant, most of the discarded green tea seeds are treated with water or a solvent again, and extract fractions having anticancer and antiinflammatory activity therefrom. Obtained.

Therefore, the present invention can provide an anti-inflammatory or anti-cancer composition containing skim green tea seed extract as an active ingredient.

In the present invention, the green tea seed can be used as long as it is a commercially available green tea seed, and preferably, the outer shell and the inner skin of the green tea seed are removed, washed, and then at a temperature of (45) to (55) ° C. 2 After drying for 1 to 4 days, the ground and pulverized can be used.

In addition, the degreasing can be used as long as it is a method for removing fat from green tea seeds known in the art, and preferably an organic solvent for degreasing. More preferably, it can be degreased by adding n-hexane solvent to the green tea seed powder, wherein the amount of the n-hexane solvent is added by 1500 ~ 2500ml of n-hexane based on 100g of green tea seed powder for 22 to 26 hours It can be degreased at room temperature.

The degreased green tea seed powder may then be added to water or an organic solvent to obtain a green tea seed extract. The addition of water or an organic solvent is about 5 to 15 times the weight of the green tea seed powder, preferably about 8 to 12. Can be added in pear quantities.

In addition, the method of obtaining an extract from the green tea seed powder degreased by adding water or an organic solvent may be performed using extraction methods such as cold sediment extraction, hot water extraction, ultrasonic extraction, reflux cooling extraction, etc. which are known in the art. It may be preferably obtained by repeating the extraction 1 to 10 times, preferably 2 to 7 times by using hot water extraction or reflux cooling extraction method for 2-6 hours at a temperature of 75 ~ 105 ℃. .

The organic solvent is not limited thereto, but may be selected from the group consisting of methanol, ethanol, propanol, isopropanol, butanol, acetone, ether, benzene, chloroform, ethyl acetate and methylene chloride.

In an embodiment of the present invention, 10 times distilled water was added to degreased green tea seed powder, and hot water was extracted for 4 hours at a temperature of 100 ° C. to obtain green tea seed extract, and 10 times ethanol was added to degreased green tea seed powder. The mixture was extracted under reflux for 4 hours at 80 ° C., followed by filtration under reduced pressure to obtain an ethanol extract. Thereafter, distilled water and petroleum ether were added to the ethanol extract to fractionate the water layer and the petroleum ether layer, and then ethyl acetate was added to the water layer. After fractionation, the mixture was partitioned into an aqueous layer and an ethyl acetate layer, and then, butanol was added to the aqueous layer and fractionated into an aqueous layer and a butanol layer. At this time, the fractionation by the solvent treatment for each step was repeated twice, and the preparation process of each fraction of the degreased green tea seed according to the embodiment is shown in FIG. 1.

On the other hand, the green tea seed extract obtained by water and solvent treatment from the green tea seed powder degreased according to the present invention as described above is characterized by having an antioxidant activity.

According to an embodiment of the present invention, the hot phenol extract, ethanol fraction, petroleum ether fraction as a result of measuring the total phenol content and the equivalent of trolox contained in the green tea seed extract obtained by the method of the present invention , Ethyl acetate fraction and butanol fractions were found to have the highest antioxidant capacity, especially ethanol fraction and hot water extract showed the highest antioxidant capacity, the ethyl acetate fraction was the highest in the total phenol content (Table 3 and See Table 4).

Therefore, according to the above results, the extract of the degreased green tea seed powder according to the present invention can prevent or treat diseases caused by oxidative stress, such as aging, hypertension, arteriosclerosis, stroke, fatty liver, cirrhosis, diabetes, cataracts, and the like. I could see that.

In addition, the extract of the skim green tea seed powder according to the present invention is characterized by excellent anti-inflammatory activity.

Inflammation, as previously described, causes tumor necrosis factor-α (TNF-α) in immune cells, such as macrophages, when harmful stimuli, such as lipopolysaccharide (LPS), pro-inflammatory factors, and irradiation, excessively overwhelm the human immune system. Inflammatory diseases caused by excessive induction of proinflammatory agents such as IL-1 (interleukin-1), IL-6, prostaglandin, luecotriene and nitric oxide (NO) It is known.

In order to investigate whether the extract of the skim green tea seed powder prepared in the present invention has anti-inflammatory activity, the present inventors investigated the inflammation-producing cytokine PGE2 and inflammation according to the treatment of skim green tea seed extract in one embodiment of the present invention. Expression of TNF-α, a transcription factor, was confirmed by Western blot. As a result, PGE2 and TNF-α were found to decrease in the amount of expression by treatment of each solvent extract (hot water extract, ethanol fraction, petroleum ether fraction, ethyl acetate fraction and butanol fraction) of degreased green tea seeds. It was found that the amount of expression was reduced (see Table 6 and FIG. 8).

Therefore, the degreased green tea seed extract according to the present invention is characterized by suppressing the expression of proinflammatory factors, thereby preventing or treating inflammatory diseases.

The type of the inflammatory disease in the present invention is not limited thereto, and may include gastritis, colitis, arthritis, nephritis and hepatitis.

Furthermore, the extract of the skim green tea seed powder according to the present invention is characterized by excellent anticancer activity.

In general, cancer is a step in which mutations are induced in DNA by carcinogens, a benign tumor appears, a benign tumor is converted into a malignant tumor, and an increase in the degree of malignancy and metastasis, It is known to proceed in a multi-step, and the treatment of such cancer is used to inhibit the cell proliferation of cancer cells or to induce apoptosis of cancer cells.

The present inventors investigated whether the extract of the skim green tea seed powder prepared in the present invention has an activity of inhibiting the cell proliferation of cancer cells, according to an embodiment of the present invention, the degreasing for each solvent obtained in the present invention Extracts of green tea seeds were treated to liver cancer cell lines, and then cell viability was measured by MTT analysis. As a result, each fraction was found to inhibit the cell proliferation of liver cancer cells, in particular, butanol fraction was found to have the highest cancer cell proliferation inhibitory activity at the lowest concentration (see Table 5 and Figure 2). .

On the other hand, enzymes involved in the metabolism of carcinogens in the body are largely divided into phase 1 enzymes and phase 2 enzymes. Phase 1 enzymes include arylhydrocarbon hydroxylase and cytochrome P450, and phase 2 enzymes include QR, HO-1, glutathione S-transferase (GST), UDP-glucuronyltransferase (Agrawal V, Huang N, Miller WL. 2008, Pharmacogenetics and Genomics . 18: 569-576, Allanson M, Reeve VE. 2004. J Invest Dermatol 122: 1030-1036, Androutsopoulos VP, Tsatsakis AM, Spandidos DA. BMC Cancer . 9: 187-204., Hart SN, Zhong XB. 2008, Expert Opin . drug Metab . Toxicol . 4: 439-452). Among them, the induction reaction of the phase 2 enzyme system acts as an important defense mechanism in vivo by detoxifying extraneous foreign substances, and the mechanism of action of the phase 2 enzyme system inhibits the reaction of highly reactive carcinogenic components with intracellular molecules. The protection at the cellular level is possible by the activation of phase 2 detoxifying enzymes and antioxidant enzymes such as HO-1 and QR. In particular, QR is an anti-tumor enzyme activity as an index enzyme of phase 2 enzyme. Widely used in the search for factors (Farombi EO, Surh YJ. 2006, J. Biochem . Mol . Biol . 39: 479-491., Fresco P, Borges F, Diniz C, Margues MPM. 2006, Med Res Rev 26: 747-766., Greten FR, Eckmann L, Gereten TF et al. 2004, Cel l 118: 285-296., Moon JY. 2004, The Korean Journal of Meridian & Acupoint 21: 139-145) .

Therefore, in another embodiment of the present invention, the extract of the degreased green tea seed prepared in the present invention is detoxified, that is, the activity of the first phase enzymes CYP1A1 and CYP1A2 and the second phase enzymes HO-1, ARE and QR The results showed that the solvent extract fractions of the extracts of defatted green tea seeds increased the expression of CYP1A2, especially in the phase 1 enzyme system (see FIG. 5), HO-1, ARE and QR. It has been shown that the activity of increases in a concentration-dependent manner (see Figs. 3 and 6).

Through the above results, the present inventors confirmed that the extract of the defatted green tea seed according to the present invention had anticancer activity through inhibiting the cell proliferation of cancer cells and promoting the activity of the detoxification toxin system. It was investigated whether the extract affects the cell cycle of cancer cells. In general, the regulation of cell cycle related to cell proliferation is known to be regulated by various genes involved in each cycle. Can be defined as a disease due to abnormalities in the cell cycle.

Therefore, cell cycle inhibition at a specific time period may be induced by decreased expression of cell cycle regulatory positive factors or overexpression of negative regulators. In particular, cell cycle inhibition in cancer cells may cause cancer cell death (Sherr CJ. 2000. The Pezcoller lecture:. cancer cell cycles revisited Cancer Res 60: 3689-3695).

Thus, in one embodiment of the present invention, each solvent extraction fractions of degreased green tea seeds according to the present invention were treated with liver cancer cell lines, and cell cycle changes of liver cancer cells were examined. As a result, the treatment concentration of the extract fractions was increased. To determine the replication time of the DNA in the cell cycle and to extend the G1 stage to prepare for the cell division of the G2 phase has been shown to cause abnormalities of the cell cycle (see Figure 7a and 7b), in particular ethyl acetate extraction Fractions and hydrothermal extracts have been shown to effectively extend the G1 phase of the cell cycle.

Therefore, the present inventors have found that each extract fraction of the degreased green tea seed according to the present invention has anticancer activity through a process leading to cell cycle abnormality of cancer cells and causing cell death.

Therefore, the present invention can provide an anti-inflammatory or anti-cancer composition comprising skim green tea seed extract as an active ingredient, the composition of the present invention can be used as a pharmaceutical composition for the prevention and treatment of inflammatory diseases or cancer.

In addition, the composition according to the present invention may comprise 0.1 to 50% by weight of the green tea seed extract degreased relative to the total weight of the composition.

Types of the cancer that can be prevented or treated by the composition of the present invention is not limited thereto, and may include liver cancer, stomach cancer, colon cancer, lung cancer, breast cancer, rectal cancer and pancreatic cancer.

The anti-inflammatory or anticancer composition according to the present invention may include a pharmaceutically effective amount of degreased green tea seed extract alone or may include one or more pharmaceutically acceptable carriers, excipients or diluents. The pharmaceutically effective amount herein refers to an amount sufficient to prevent, ameliorate and treat an inflammatory disease or cancer condition.

The pharmaceutically effective amount of the skim green tea seed extract according to the present invention is 0.5 to 100 mg / day / kg body weight, preferably 0.5 to 5 mg / day / kg body weight. However, the pharmaceutically effective amount may be appropriately changed according to the degree of inflammatory disease or cancer symptoms, the age, weight, health condition, sex, route of administration and treatment period of the patient.

In addition, the pharmaceutically acceptable refers to a composition that is physiologically acceptable and does not cause an allergic reaction such as gastrointestinal disorders, dizziness or the like when administered to humans. Examples of such carriers, excipients and diluents include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, Polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. In addition, fillers, anti-coagulants, lubricants, wetting agents, fragrances, emulsifiers and preservatives may be further included.

In addition, the compositions of the present invention may be formulated using methods known in the art so as to provide rapid, sustained or delayed release of the active ingredient after administration to the mammal. The formulations may be in the form of powders, granules, tablets, emulsions, syrups, aerosols, soft or hard gelatine capsules, sterile injectable solutions, sterile powders.

The anti-inflammatory or anticancer composition according to the present invention can be administered through various routes including oral, transdermal, subcutaneous, intravenous or intramuscular, and the dosage of the active ingredient is determined by the route of administration, the age, sex, weight of the patient and the severity of the patient. It may be appropriately selected depending on several factors. In addition, the anti-inflammatory or anticancer composition of the present invention can be administered in parallel with known compounds having the effect of preventing, ameliorating or treating inflammatory diseases or cancer symptoms.

Therefore, the present invention can provide a medicament for the prevention and treatment of inflammatory diseases or cancer symptoms, including a composition containing skim green tea seed extract as an active ingredient.

Furthermore, the anti-inflammatory or anti-cancer composition according to the present invention not only provides an effect of alleviating inflammatory diseases and cancer symptoms through excellent antioxidant, anti-inflammatory and anti-cancer effects, and also has no toxicity and side effects on drugs, so it is safe even when taken for a long time. Green tea that can be used as a raw material of the extract used in the present invention is a natural plant that can be used as food grade, and thus the composition of the present invention has a very stable feature for the body.

Therefore, the anti-inflammatory or anti-cancer composition of the present invention can be added to food for the purpose of preventing and improving inflammatory diseases or cancer symptoms, the composition of the present invention as a composition for food for the prevention and improvement of inflammatory diseases or cancer symptoms Can be used.

Therefore, the composition of the present invention can be easily utilized as a food, such as a main raw material, side ingredients, food additives, functional foods or beverages that are effective in preventing and improving inflammatory diseases or cancer symptoms.

As used herein, the term “food” refers to a natural product or processed product containing one or more nutrients, and preferably means a state in which it can be directly eaten through a certain processing step, It includes all foods, food additives, functional foods and drinks.

Foods to which the anti-inflammatory or anticancer composition according to the present invention can be added include, for example, various foods, beverages, gums, teas, vitamin complexes, functional foods, and the like. In addition, in the present invention, food includes special nutritional products (e.g., formulated milk, young, infant food, etc.), processed meat products, fish products, tofu, jelly, noodles (e.g. ramen, noodles, etc.), bread, health supplements, seasonings. Foods (e.g. soy sauce, miso, red pepper paste, mixed soy sauce), sauces, confectionery (e.g. snacks), candy, chocolates, gums, ice creams, dairy products (e.g. fermented milk, cheese, etc.), other processed foods, kimchi, Pickled foods (various kimchi, pickles, etc.), beverages (e.g., fruit drinks, vegetable drinks, soy milk, fermented beverages, etc.), natural seasonings (e.g. ramen soup, etc.) are not limited thereto. The food, beverage or food additive may be prepared by a conventional production method.

In addition, the term "functional food" refers to the control of biological defense rhythms and disease prevention of food groups or food compositions that have added value to the food by using physical, biochemical, or biotechnological techniques to act and express the function of the food for a specific purpose. It means a food that is designed and processed to fully express the body's regulatory function regarding recovery and the like, and specifically, it may be a health functional food. The functional food may include food acceptable food additives, and may further include appropriate carriers, excipients and diluents commonly used in the manufacture of functional foods.

In addition, in the present invention, the "beverage" refers to a generic term for drinking to quench thirst or to enjoy a taste and includes a functional drink. The beverage is an essential ingredient in the ratio indicated, and includes the composition for the prevention and amelioration of the inflammatory disease or cancer symptom, and there is no particular limitation on the other ingredients. It may contain as.

Furthermore, foods containing the composition of the present invention, in addition to those described above, include various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavoring agents, colorants and fillers (such as cheese and chocolate), and pectic acid. And salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like, which components may be independently or in combination Can be used.

In the food containing the anti-inflammatory or anti-cancer composition of the present invention, the amount of the composition according to the present invention may be included in 0.001% to 90% by weight of the total food weight, preferably 0.1% to 40% by weight It may include, in the case of a beverage, may be included in a ratio of 0.001g to 2g, preferably 0.01g to 0.1g based on 100ml, but for long-term intake of health and hygiene or health control In this case, it may be less than the above range, and the active ingredient is not limited to the above range because it may be used in an amount above the above range because there is no problem in terms of safety.

Therefore, the present invention can provide a health functional food for the prevention or improvement of inflammatory diseases or cancer diseases containing skim green tea seed extract according to the present invention as an active ingredient, but the form of the food is not limited thereto. It may be in powder, granule, tablet, capsule or beverage form.

Hereinafter, the present invention will be described in detail with reference to the following examples and experimental examples.

However, the following Examples and Experimental Examples are merely illustrative of the present invention, and the contents of the present invention are not limited by the following Experimental Examples.

< Reference Example  1>

Cell culture conditions, reagents and statistical processing methods used in the Examples of the present invention were performed as described below.

Cell Culture and Reagents

HepG2 cells were purchased from the American Type Culture Collection (ATCC), used, cultured in Dulbecco's modified eagle media (DMEM) medium with 10% Fetal bovine serum (FBS), and passaged once every 3 to 4 days. Cultured in a medium, and cell culture was performed under conditions of 37 ° C. and 5% CO ₂ in a CO ₂ incubator.

Statistical processing

The analysis results of the experiments performed in the examples of the present invention are expressed as mean ± standard error (SE), and the significance is Duncans using one-way ANOVA of the SPSS / PC + package. The analysis was performed by Duncan's multiple test.

< Example  1>

Degreased Green tea seed  Preparation of Extract

<1-1> degreased Green tea seed  Manufacture of powder

Green tea seed was purchased from Korea (Korea) and used. After removing the outer shell and the inner skin, wash it clean, dry it for 3 days by hot air drying at a temperature of 50 ℃, and then powder it into 40 mesh. N-hexane was added to degrease for 24 hours, filtered using a vacuum filter (Whatmann No. 1), washed once with n-hexane and re-filtered. Thereafter, the resultant was dried for 48 hours to obtain skim green tea seed powder having a recovery of 86.49%.

<1-2> degreased Green tea seed Hydrothermal  Preparation of Extracts and Ethanol Extracts

10 times distilled water of the powder was added to the degreased green tea seed powder prepared in <1-1>, and reflux extraction was performed at 100 ° C. for 4 hours, followed by centrifugation (1,500 rpm, 15 minutes). Filtration under reduced pressure, followed by freeze-drying to obtain a hydrothermal extract of skim green tea seed. In addition, ethanol extract was obtained by adding ethanol corresponding to 10 times the degreased green tea seed powder, and reflux extraction at 80 ° C. for 4 hours, followed by filtration under reduced pressure, followed by drying using an evaporator. At this time, the recoveries of the hydrothermal extract and ethanol extract of the green tea seed degreased through the above process are as described in Table 1 below.

Recovery Rate of Ethanol Extracts and Hot Water Extracts of Defatted Green Tea Seeds DGTS Hot water 35.84% Ethanol 19.94%

DGTS; defatted green tea seed

<1-3> Preparation of fraction extract of degreased green tea seed The fraction extract of degreased green tea seed was prepared by the following method, that is, 10 g of ethanol extract (EtOH) prepared in <1-2> was distilled water 500 After dissolving by adding mL, 500 mL of petroleum ether (PE) was added and mixed, and then the supernatant, which is a PE layer, was recovered. The extraction was repeated twice (PE fraction). 500 mL of ethyl acetate (EtOAC) was added to the remaining water layer. After mixing, the ethyl acetate layer as a supernatant was recovered, and the reaction was repeated twice to extract the ethyl acetate fraction. Thereafter, 500 mL of butanol (BuOH) was added to the remaining water layer, followed by mixing. The butanol layer, which was the supernatant, was recovered. The process for preparing a fraction extract of the green tea seed degreased from the ethanol extract of the skim green tea seed as described above is shown in FIG. In addition, the final water fraction was freeze-dried, all the remaining fractions were dried using an evaporator and used as a sample for the next experiment, the yield of each fraction is as shown in Table 2 below.

Recovery of Defatted Green Tea Seed Extract DGTS Ethanol extract        10.0 g    PE 0.115 g    EtOAC 0.144 g    BuOH 4.059 g H ₂ O 4.952g

DGTS; defatted green tea seed

< Example  2>

Degreased Green tea seed  Extract Antioxidant activity  analysis

Antioxidant activity of the skim green tea seed extract according to the present invention prepared in Example 1 was confirmed by measuring the total phenol content and trolox equivalent of the extract, that is, fraction in Example <1-3> Total phenol content of each sample Measured by the Folin-Dennis method (AOAC. 1985. Official method of analysis of the Association of Official Analytical Chemists. 16th ed. Chemists, Washington, DC), for this purpose, 200 mL of each extract, 1 mL of 50% Folin-Ciocalteu reagent and 7.5% 800 μL of Na ₂ CO ₃ solution was added thereto, and the absorbance was measured at 760 nm for 45 minutes in the dark. Gallic acid was used as a standard.

In addition, the measurement of total antioxidant activity was performed by Erel's method (Erel C et al. 2004, A novel automated) with trolox equivalent antioxidant capacity (TAEC) for the extracts and fractions of <1-2> and <1-3>. method to measure total antioxidant response against potent free radical reaction. Clinical Biochem 37, using 112-119.) 2,2'-azino-bis- (3-ethybenzthiazoline -6-sulfonic acid (ABTS, Sigma) radical cation decolorization analysis was investigated through. then, the manufacturing of 7mM ABTS solution TAEC is the ABTS ⁺ the stock solution was prepared and left in then a solution of the 2.45 mM potassium persulfate cow for 16 hours. Thereafter, the ABTS ⁺ the stock solution of PBS solution (5 mM, pH 7.4), the absorbance was adjusted to 0.70 at 734 nm, and 50 μL of each sample was added and mixed. The absorbance was measured at 30 ° C. after exactly 1 minute and 6 minutes. Each sample was prepared and used. Antioxidant Activity was expressed as trolox equivalent. In addition, the statistical analysis was performed using SPSS software soup All results were tested for significance at p <0.05 were expressed as mean ± SE, among groups with ANOVA and Duncan's multiple range test.

As a result, as shown in Table 3, the antioxidant capacity of the skim green tea seed extract, each extract fraction was shown to have an antioxidant capacity, among which the hot water extract (HW) showed the highest antioxidant capacity, The ethanol fraction and butanol fraction showed the next highest antioxidant capacity, while the petroleum ether layer showed the lowest antioxidant capacity.

In addition, as a result of measuring the content of total phenol per 100 mg of each extract, as shown in Table 4, the total phenol ethyl acetate fraction was found to be the highest as 3.197 ± 0.195 mg, while 0.250 ± 0.043 in the petroleum ether layer The lowest level was shown in mg.

Total Antioxidant Activity of Degreased Green Tea Seed Extract (Trolox equivalent (mM)) Extract DGTS EtOH 4.74 PE 1.26 EtOAC 3.65 BuOH 4.53 H ₂ O 3.68 HW (Hot Water Extract) 4.92

GTS; defatted green tea seed

Total phenolic content (gallic acid) DGTS
(mg / 100 mg extracts) EtOH 1.315 ± 0.009 ^d PE 0.250 ± 0.043 ^e EtOAC 3.197 ± 0.195 ^a BuOH 2.132 ± 0.011 ^c H ₂ O 0.903 ± 0.041 ^d HW (Hot Water Extract) 2.669 ± 0.031 ^b

GTS; defatted green tea seed

< Example  3>

Degreased Green tea seed  Anticancer Activity Analysis of Extracts

The inventors of the present invention fractionated green tea seed fractions according to the present invention fractionated in Example 1 to investigate whether there is anti-cancer activity in HepG2 cells in the cancer cell proliferation inhibitory effect, the effect on detoxifying enzyme activity and cell cycle The effect on activity was analyzed respectively. HepG2 cell culture conditions and the treatment of the sample for the above analysis was treated in the following manner. First, HepG2 cells were cultured in DMEM (10% FBS) medium, and cell culture was performed under conditions of 37 ° C. and 5% CO ₂ in a CO ₂ incubator. The sample treatment of skim green tea seed extract was 2.5, 5, 10, 20 ㎍ / mL for ethanol extract fraction, 50, 100, 150, 200 ㎍ / mL for petroleum ether extract fraction, and ethyl acetate extract fraction. , 10, 20, 30, 40, 50 μg / mL, butanol extract fraction, 1, 2.5, 5, 7.5, 10 μg / mL, water fraction, 25, 50, 100 μg / mL, hydrothermal extract In this case, the cells were treated with 20, 30, 40, and 50 μg / mL, respectively, and then cultured for 48 hours to confirm the final concentration as cytotoxic. Final concentrations of ethanol extract 2.5, 5, 10, 20 μg / mL, petroleum ether extract 50, 100, 150 μg / mL, ethyl acetate extract 10, 20, 30 μg / mL, butanol extract 1, 2.5, 5 μg / mL , Water extract 25, 50, 100 ㎍ / mL, hot water extract was determined to be 20, 30, 40, 50 ㎍ / mL.

<3-1> Effect of Cancer Cell Proliferation

The anticancer activity of the degreased green tea seed extract fraction according to the present invention was analyzed by measuring the effect of inhibiting cancer cell proliferation, that is, the effect of each extract fraction on the proliferation of cells through the MTT assay. The MTT assay dispensed HepG2 cells into 96 well plates at 5 × 10 ³ cells / well (200 μL), followed by preincubation for 24 hours, and degreased green tea seed fractions as described above at each concentration. After administration, 48 hours after, the viability of the cells was measured, and the results are shown in Table 5 and FIG. 2.

Cytotoxic IC ₅₀ Value (μg) of Green Tea Seed Extract Degreased in HepG2 Cell Line EtOH PE EtOAC BuOH H2O HW IC ₅₀ value 6.58 120.77 18.29 3.54 44.37 25.02

As a result, as shown in Figure 2, the defatted green tea seed extract fraction of the present invention was found to have the activity of inhibiting the cell proliferation of HepG2 cells, liver cancer cells, IC ₅₀ value of the liver cancer cell inhibitory ability of the green tea seed extract As shown in Table 5, butanol layer showed the lowest value of 3.54 μg, followed by ethanol layer (6.58 μg), ethyl acetate layer (18.29 μg), hot water extract (25.02 μg), water fraction layer ( 44.37 μg) and petroleum ether layer (120.77 μg).

<3-2> Detoxification system  Active measurement

In order to investigate the effect of deionized toxin-based activity on the hot water extract and the solvent extract fraction of the skim green tea seed extract according to the present invention, NADPH (quinone reductase: QR) activity analysis, Western blot and reporter gene activity analysis was performed It was.

First, NADPH (quinone reductase: QR) activity assay, such as Benson (Benson AM, Hunkeler MJ, Talalay P. 1980. Increase of NAD (P) H: quinone reductase by dietary antioxidants:. Possible role in protection against carcinogenesis and toxicity Proc Natl Acad Sci USA 77: 5216-5220.), I.e., 1 × 10 ⁶ HepG2 cells were dispensed into a cell culture dish (φ100 mm) and cultured 24 hours before, followed by α-MEM (10% FBS). Removed and incubated for 48 hours by adding the green tea seed extract fraction according to the invention to a fresh medium. The medium was removed and then washed twice with cold PBS (137 mM NaCl, 2.7 mM KCl, 10 mM Na ₂ HPO ₄ , 2 mM KH ₂ PO ₄ ) and collected with 0.25M cross. The collected cells were homogenized with an ultrasonic cell digester (50W, Kontes, Vineland, NJ, USA), and then centrifuged (10,500 × g, 15 minutes) using a micro centrifuge (VS-15000CFN11, Vision, Seoul, Korea). Supernatant was obtained. To 200 μL of cell supernatant, 250 mM tris-HCl (pH 7.4), 70 mg bovine serum albumin, 0.1% Tween 20, 500 uM FAD, 2 mM NADH was added, and the substrate 2,6-dichlorophenolindophenol (DCPIP) was added to the protein. The extent of oxidation for 1 minute per mg was measured at 600 nm. Protein quantitation was performed by the Bradford method. In addition, the QR activity used in the present invention is widely used in the search for anti-tumor enzyme-based activator as an indicator enzyme of a second phase enzyme system of a certain substance.

In addition, the Western blot collected HepG2 cells treated with various concentrations of the extraction fraction as described above, quantitate the protein according to the Bredford method, and then electrophoresed 30 ㎍ of the protein by SDS-PAGE, After transferring the proteins to a PVDF membrane (polyvinylidene fluoride; pore size 0.45 μm, Millipore), the PVDF membrane was transferred to TBS / T (Tris buffered saline-Tween; 200 mM Tris, 1.37 M NaCl) containing 5% non-fat dry milk. , pH 7.6, 0.1% Tween 20) was reacted overnight to block nonspecific proteins. Thereafter, the reaction was performed for 3 hours with antibodies against CYP1A1, CYP1A2, and HO-1, and then for 2 hours with anti-rabbit IgG or anti-goat IgG, which is a secondary antibody to each antibody. Each reaction was washed three times with TBS / T for 10 minutes. After the final wash, the corresponding protein bands for each antibody were expressed using a Supersignal west pico chemiluminescent substrate (Pierce, Rockford, IL, USA). 4 is shown.

In addition, the reporter gene activity assay was performed through ARE-Luciferase assay, for this purpose first dispense 1 × 10 ⁵ HepG2 cells / 60 mm dish per plate (φ60 mm), then pre-incubate the cells for 24 hours and , Ethanol extract fraction 2.5, 5, 10, 20 μg / mL, petroleum ether extract fraction 50, 100, 150 μg / mL, ethyl acetate extract fraction 10, 20, 30 μg / mL according to the present invention, butanol extract fraction 1, 2.5, 5 ㎍ / mL, water extract fractions 25, 50, 100 ㎍ / mL and hot water extracts were treated to 20, 30, 40, 50 ㎍ / mL, respectively, and then cultured for 48 hours, and then cells were collected to obtain luciferase activity. Was measured according to the Promega E4030 method. That is, the collected samples were analyzed using a luminometer (Perkin Elmer Victor2 1420), and the enzyme activity of the analyzed samples was corrected to specific activity values by quantifying protein content according to the Bredford method, and the results are shown in FIG. 5. Indicated.

As a result of the experiment, the QR activity of the degreased green tea seed extract in HepG2 cells was found to increase significantly compared to the control when treated with a concentration of 20 μg in the ethanol layer, as shown in FIG. In the case of, butanol layer, the treatment increased more than 2.5 μg compared to the control group, while the petroleum ether layer and ethyl acetate showed a tendency to increase slightly compared to the control group, but there was no significant difference. In addition, the water extract fraction and hot water extract were found to decrease rather at high concentrations, and the ethanol layer and butanol layer showed the best QR activity among the fraction extracts of degreased green tea seeds.

In addition, the measurement results of CYP1A1 and CYP1A2 protein expression by Western blot, as shown in Figure 4, in each extract fraction of degreased green tea seed, butanol layer was found to increase the gene expression of CYP1A1, while in other fractions The expression of CYP1A2 was found to be increased in all fractional extracts, especially in the ethanol and butanol layers. Further petroleum ether layer was shown to increase the protein expression of HO-1 (see Figure 5).

In addition, as a result of the reporter analysis, in the case of the ethanol extract fraction layer, petroleum ether layer and ethyl acetate layer of the skim green tea seed according to the present invention, the activity of ARE was increased in a concentration-dependent manner.

<3-3> Analysis of the effect on the cell cycle

The effect of green tea seed fraction extract on cell cycle distribution was investigated to investigate whether the growth factor of HepG2 hepatocellular carcinoma cells by the green tea seed fraction extract was associated with the perturbation of cell cycle. Cell cycle irradiation was performed using propidium iodide (PI) staining. 100 μL of PBS and 200 μL of ethanol were added to the collected cells, and the cells were fixed at 4 ° C. for 1 hour. After washing twice with PBS, 50 μg / mL RNaseA (1.12% sodium citrate buffer) was added 250 μL and reacted at 37 ° C. for 30 minutes, and 50 μg / mL PI (1.12% sodium citrate buffer) was added 250 μL. After reaction at 37 ° C. in the dark for 30 minutes, cell cycle ratios were analyzed by flow cytometry (Becton Dickinson, FACS Calibur, NJ, USA).

As a result, as shown in Figures 7a and 7b, the ethyl acetate layer and hot water extract of the skim green tea seed extract was shown to extend the sub G1 of the cell cycle as the treatment concentration increases. In general, the G1 phase of the cell cycle mainly determines the time of DNA replication. Afterwards, cell division is prepared in the G2 phase and the time of initiation of division is determined. In the cell cycle, G1, especially sub G1, plays a decisive role in the survival of cells. In turn, it is known to induce cell death by ultimately leading to abnormal cell cycles.

Therefore, it can be seen from the above results that the degreased green tea seed extracts according to the present invention have anticancer activity by inducing apoptosis of cancer cells by extending the period of sub G1, particularly in the cell cycle of liver cancer cells.

< Example  4>

Degreased Green tea seed  Anti-inflammatory Activity Analysis of Extracts

In Example 3, it was found that the degreased green tea seed extract has anti-cancer activity, and the present inventors further confirmed that the degreased green tea seed extract has an anti-inflammatory effect, which is an inflammation-related protein PGE2 (inflammatory cytokine). Cain) and inflammatory transcription factor TNF-a expression was analyzed by Western blot.

To this end, in Example 3, the total protein was isolated from HepG2 cells, the liver cancer cell line treated with the extract fractions of each concentration, wherein the separation process was performed by PROPREP ^™ Protein Extract Solution (iNtRON Biotechnology). That is, remove the culture medium from the cell culture plate of 100 mm dish (5 × 10 ⁶ cells / dish), wash twice with pre-prepared cold PBS (pH 7.4), and then add 1 mL cold PBS (pH 7.4) to the cells. Was collected. Thereafter, the upper layer was removed by centrifugation (12,000 rpm, 20 seconds) at 4 ° C, 400 μL proprep was added for cell lysis, and incubated at -20 ° C for 20 minutes, followed by centrifugation at 4 ° C (12,000 rpm, 5 minutes). Cell extract supernatant (total protein) was obtained. The obtained protein was quantified or stored at -20 ° C for analysis.

In addition, Western blot quantified the obtained cell-extracted supernatant by Bradford method to determine the protein concentration, and then electrophoresed 30 ㎍ of protein on 10% SDS-PAGE, and then transferred to the nitrocellulose membrane. The membrane was then blocked with 5% blocking solution (5% BSA, 5% nonfat dried milk in TBST) for 2 hours as long as washed with TBST [10 mM Tris-HCl (pH 8.0), 150 mM NaCl, 0.05% Tween 20]. Later, primary antibodies (rabbit anti-mouse iNOS antibody, polyclonal rabbit COX-2 antibody, Cell Signaling, Phospho-IκBα rabbit mAb, Cell Signaling, Phospho-p65 rabbit mAb, Cell Signaling, polyclonal Actin antibody and Santa Cruz Biotechnology) were reacted overnight at 1: 1,000, then rinsed three times with TBST and diluted goat anti-mouse immunoglobulin G (IgG) secondary antibody bound with alkaline phosphatase at 1: 2,000 for 1 hour. Reaction at room temperature. Subsequently, color development with 5-bromo-4-chloro-3-indolyl phosphate / nitroblue tetrazolium color development substrate (Promega) or 20X LumiGLO (R) reagent and 20X Peroxide (Cell Signaling) on an X-ray film (Amersham Hyperfilm ^TM ECL) Was developed by emitting light. In addition, statistical analysis was performed using SPSS software and all results were expressed as mean ± SE, and the significance was tested at p <0.05 by one-way ANOVA test, and the results are shown in Table 6 and FIG. 8.

Result of PGE2 Expression Level in HepG2 Cells Treated with Green Tea Seed Extract μg / mL PGE2 (pg / mL) EtOH 0 38.9 ± 0.0 ^{1) a2 )} 2.5 -214.1 ± 34.0 ^bc 5 -154.6 ± 13.5 ^b 10 -248.6 ± 29.5 ^c 20 -180.6 ± 23.5 ^bc PE 0 38.9 ± 0.0 ^a 50 -150.1 ± 0.0 ^b 100 -206.1 ± 58.0 ^b 150 -212.1 ± 55.0 ^b EtOAC 0 38.9 ± 0.0 ^a 10 -152.6 ± 49.5 ^b 20 -131.1 ± 0.0 ^b 30 -128.1 ± 48.0 ^b BuOH 0 38.9 ± 0.0 ^a One -165.1 ± 0.0 ^b 2.5 -196.1 ± 10.0 ^b 5 -203.6 ± 31.5 ^b H ₂ O 0 38.9 ± 0.0 ^a 25 -185.1 ± 28.0 ^bc 50 -256.6 ± 63.5 ^c 100 -51.1 ± 33.0 ^ab HW 0 38.9 ± 0.0 ^a 20 -220.1 ± 47.0 ^b 30 -212.1 ± 15.0 ^b 40 -351.1 ± 0.0 ^c

¹⁾ mean ± SE ²⁾ ANOVA and Duncan's multiple test at p <0.05

As a result, as shown in Table 6, it was shown that each extract fraction and hot water extract of the degreased green tea seed according to the present invention in HepG2 cells, which are liver cancer cells, inhibit the production of PGE2 in cells, and inflammatory transcription factors. The expression of phosphorus TNF-α was also shown to decrease in a concentration dependent manner (see FIG. 8).

Furthermore, the formulation examples of the pharmaceutical composition comprising the extract of the present invention, but the present invention is not intended to limit it, but is intended to explain in detail only.

< Formulation example  1> Powder  Produce

Degreased Green Tea Seed Extract 300 mg

Lactose 100 mg

Talc 10 mg

The above ingredients are mixed and filled in an airtight cloth to prepare a powder.

< Formulation example  2> Preparation of tablets

Degreased Green Tea Seed Extract 50 mg

Corn starch 100 mg

Lactose 100 mg

2 mg magnesium stearate

After mixing the above components, tablets are prepared by tableting according to a conventional method for preparing tablets.

< Formulation example  3> Preparation of capsule

Degreased Green Tea Seed Extract 50 mg

Corn starch 100 mg

Lactose 100 mg

2 mg magnesium stearate

The above components are mixed according to a conventional capsule preparation method and filled in gelatin capsules to prepare capsules.

< Formulation example  4> Preparation of injection

Degreased Green Tea Seed Extract 50 mg

Appropriate sterile distilled water for injection

pH adjuster

(2 ml) per 1 ampoule according to the usual injection preparation method.

< Formulation example  5> Liquid  Produce

Degreased Green Tea Seed Extract 100 mg

10 g of isomerized sugar

5 g of mannitol

Purified water

According to the conventional method of preparing a liquid solution, each component is added to the purified water to dissolve, the lemon flavor is appropriately added, the above components are mixed, purified water is added, the whole is adjusted to 100 ml by the addition of purified water, and then concentrated in a brown bottle. The solution is prepared by sterilization.

< Formulation example  6> Manufacture of health food

Defatted Green Tea Seed Extract 1000 mg

Vitamin mixture proper amount

70 μg of Vitamin A Acetate

Vitamin E 1.0 mg

Vitamin B1 0.13 mg

Vitamin B2 0.15 mg

Vitamin B6 0.5 mg

0.2 μg of vitamin B12

Vitamin C 10 mg

10 μg biotin

Nicotinic Acid 1.7 mg

Folate 50 ㎍

Calcium Pantothenate 0.5mg

Mineral mixture

Ferrous Sulfate 1.75 mg

Zinc Oxide 0.82 mg

Magnesium carbonate 25.3 mg

Potassium monophosphate 15 mg

Dibasic calcium phosphate 55 mg

Potassium Citrate 90 mg

Calcium Carbonate 100 mg

Magnesium chloride 24.8 mg

The composition ratio of the above-mentioned vitamin and mineral mixture is mixed with a composition suitable for a health food in a preferred embodiment, but the compounding ratio may be arbitrarily modified. The granules may be prepared and used for preparing a health food composition according to a conventional method.

< Formulation example  7> Health drink  Produce

Defatted Green Tea Seed Extract 1000 mg

Citric acid 1000 mg

100 g oligosaccharides

Plum concentrate 2 g

1 g of taurine

Add 900 ml of purified water

After mixing the above components according to a conventional healthy beverage manufacturing method, and then stirred and heated at 85 ℃ for about 1 hour, the resulting solution is filtered and obtained in a sterilized 2 L container, sealed and sterilized and then refrigerated and stored in the present invention For the preparation of healthy beverage compositions.

Although the compositional ratio is relatively mixed with a component suitable for a favorite drink, it is also possible to arbitrarily modify the compounding ratio according to the regional or national preference such as the demand class, the demanding country, and the use purpose.

So far I looked at the center of the preferred embodiment for the present invention. Those skilled in the art will appreciate that the present invention can be implemented in a modified form without departing from the essential features of the present invention. Therefore, the disclosed embodiments should be considered in an illustrative rather than a restrictive sense. The scope of the present invention is shown in the claims rather than the foregoing description, and all differences within the scope will be construed as being included in the present invention.

Claims (9)

Anti-inflammatory or anti-cancer composition comprising skim green tea seed extract as an active ingredient. The method of claim 1,
The degreasing anti-inflammatory or anticancer composition, characterized in that the degreasing at room temperature for 22 to 26 hours by adding 1500 ~ 2500ml n-hexane based on 100g green tea seed powder. The method of claim 1,
The extract is an anti-inflammatory or anticancer composition, characterized in that the solvent extract solubilized by adding water or an organic solvent to the degreased green tea seed powder. The method of claim 3,
The organic solvent is anti-inflammatory or anticancer composition, characterized in that selected from the group consisting of methanol, ethanol, propanol, isopropanol, butanol, acetone, ether, benzene, chloroform, ethyl acetate and methylene chloride. The method of claim 1,
The anti-inflammation or anti-cancer composition characterized in that it can prevent or treat an inflammatory disease selected from the group consisting of gastritis, colitis, arthritis, nephritis and hepatitis. The method of claim 1,
The anticancer is an anti-inflammatory or anticancer composition, characterized in that it can prevent or treat a cancer disease selected from the group consisting of liver cancer, stomach cancer, colon cancer, lung cancer, breast cancer, rectal cancer and pancreatic cancer. The method of claim 1,
An anti-inflammatory or anticancer composition comprising the extract in an amount of 0.1 to 50% by weight based on the total weight of the composition. Health functional food for the prevention or improvement of inflammatory diseases or cancer diseases containing skim green tea seed extract as an active ingredient. The method of claim 8,
The food is a health functional food for the prevention or improvement of inflammatory diseases or cancer diseases, characterized in that the powder, granules, tablets, capsules or beverage form.

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