KR20140060158A - Composition comprising roasted cucumis melo l. var makuwa makino seed extract for treating or preventing cancer - Google Patents

Abstract

The present invention relates to a composition comprising roasted Cucumis melo L. var makuwa makino seed extract as an active ingredient for preventing or treating cancer. The roasted Cucumis melo L. var makuwa makino seed extract according to the present invention has an excellent anti-cancer activity for a variety of cell strains, thereby being valuably used for preventing or treating cancer.

Description

The present invention relates to a composition for prevention or treatment of cancer comprising roasted melon seed extract as an active ingredient.

The present invention relates to a composition for preventing or treating cancer comprising roasted melon seed extract as an active ingredient.

Although the interest in health has been heightened by the improvement of quality of life, the increase of the obesity population and the chronic diseases such as cardiovascular diseases are increasing due to excessive Westernized food culture and eating habits. In particular, despite the development of modern medicine, the cause and the mortality rate of cancer continues to increase without any apparent cause. It is known that various causes such as genetic factors, immunological factors, hormonal factors, personal stress, tobacco, occupational carcinogens, and environmental pollution are known to be related to known cancer development (Kim, 1998, Mclintire and Cioppa, 1984). Recently, many efforts have been made to find a substance that can prevent this cancer in foods (Jin and Song, 2012; Murakami and Ohigashi, 2007), and many researchers have found that a new natural product- (Chang and Yoo, 2006; Isabelle et al., 2008). In this study, we investigated the effects of natural products on the development of functional foods. In recent years, as the importance of seeds has increased, research on the physiological activity of seeds has been recently reported, not only as nutritional value of seeds but also as a functional resource that can be used for prevention and treatment of diseases.

On the other hand, melon (Cucumis melo L. var makuwa Makino) has been cultivated mainly in Central Asia, China, Korea and Japan. In Korea, it is estimated that melons have been cultivated before the Silla period, and various varieties of native varieties were cultivated in each province. Melon produced as a typical fruit in summer has a higher food value because it has more calories and vitamins than other fruits and vegetables. The pharmacological action is known to be effective in the Jinhae, the gerbil action, the laxative action, the diuretic action and the constipation. However, the research on the anticancer activity of melon has not been studied until now, and the anticancer activity of melon seed is not known at all.

Accordingly, the inventors of the present invention conducted studies to develop a new anti-cancer drug, and as a result, confirmed that the roasted melon seed extract has excellent anticancer activity, thereby completing the present invention.

It is an object of the present invention to provide a pharmaceutical composition for preventing or treating cancer comprising roasted melon seed extract as an active ingredient.

It is still another object of the present invention to provide a food composition for preventing or ameliorating cancer comprising roasted melon seed extract as an active ingredient.

In order to achieve the above object, the present invention provides a pharmaceutical composition for preventing or treating cancer comprising roasted melon seed extract as an active ingredient.

The present invention also provides a food composition for preventing or ameliorating cancer comprising roasted melon seed extract as an active ingredient.

The roasted melon seed extract according to the present invention has excellent anticancer activity in various cancer cell lines and can be usefully used for prevention or treatment of cancer.

FIG. 1 is a graph showing the results of a comparison between the extracts of general melon seeds (H-Raw (hexane extract), E-Raw (ethanol extract)) and roasted melon seed extract (H- )) Was confirmed by crystal violet assay.
FIG. 2 is a view showing the anticancer activity of roasted melon seed extract (water, hexane or ethanol extract) in human liver cancer cell line (HepG2) through MTT assay.
FIG. 3 is a view showing the anticancer activity of roasted melon seed extract (water, hexane or ethanol extract) in human breast cancer cell line (MCF-7) through MTT assay.
FIG. 4 is a view showing the anticancer activity of roasted melon seed extract (water, hexane or ethanol extract) in human lung cancer cell line (A549) through MTT assay.
FIG. 5 is a graph showing the anticancer activity of roasted melon seed extract (water, hexane or ethanol extract) in human colon cancer cell line (HT-29) through MTT assay.
FIG. 6 shows the anticancer activity of roasted melon seed extract (water, hexane or ethanol extract) in human gastric cancer cell line (AGS) through MTT assay.
FIG. 7 shows the anticancer activity of roasted melon seed ethanol extracts in various cancer cell lines through MTT assay.
FIG. 8 is a graph showing the anticancer activity of roasted melon seed fraction (ethyl acetate or butanol fraction) in human liver cancer cell line (HepG2) through MTT assay.
FIG. 9 shows the anticancer activity of the roasted melon seed fraction (ethyl acetate or butanol fraction) in human breast cancer cell line (MCF-7) through MTT assay.

The present invention provides a composition for preventing or treating cancer comprising roasted melon seed extract as an active ingredient.

The composition comprises a pharmaceutical composition or a food composition.

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

Roasted melon seed extract which is an active ingredient in the composition of the present invention can be obtained by the following method.

First, the melon seeds are washed with water to remove foreign matter, and then dried. Melon seeds can be cultivated or marketed without restriction. The dried melon seeds are roasted at 100 to 300 ° C for 30 minutes to 10 hours and then pulverized. A suitable amount of the solvent is added to the pulverized roasted melon seeds so as to be completely immersed in the roasted melon seeds to obtain a roasted melon seed extract. The extraction method can be impregnated or warmed at room temperature. The extraction solvent is not limited thereto, but at least one solvent selected from water, alcohols having 1 to 4 carbon atoms, and a mixed solvent thereof may be used, and ethanol is preferably used. The roasted melon seed extract may be sequentially fractionated with an organic solvent to obtain a roasted melon seed fraction. The organic solvent may be any solvent commonly used in the art, including, but not limited to, hexane, chloroform, ethyl Acetate, butanol and the like, preferably ethyl acetate or butanol. The extract is filtered and concentrated to give a final roasted apple melon seed extract or its fractions.

The roasted melon seed extract according to the present invention has excellent anticancer activity in various cancer cell lines and can be used as medicines or health functional foods useful for the prevention or treatment of cancer.

The cancer is not limited to gastric cancer, colon cancer, breast cancer, lung cancer, liver cancer, non-small cell lung cancer, bone cancer, pancreatic cancer, skin cancer, head or neck cancer, skin or intraocular melanoma, uterine cancer, ovarian cancer Cancer of the endometrium, cancer of the cervix, vaginal cancer, vulvar carcinoma, Hodgkin's disease, esophageal cancer, small bowel cancer, lymphadenocarcinoma, bladder cancer, gallbladder cancer, endocrine cancer, endometrial carcinoma, Neoplasms, renal cell carcinomas, renal pelvic carcinomas, hematologic cancers, brain cancers, central nervous system (CNS) neoplasms, pancreatic carcinomas, pancreatic carcinomas, pancreatic carcinomas, pancreatic carcinomas, Tumor, primary central nervous system lymphoma, spinal cord tumor, brainstem glioma, or pituitary adenoma, preferably breast cancer, stomach cancer, colon cancer, lung cancer and liver cancer.

The composition of the present invention may contain at least one known active ingredient having a therapeutic effect on cancer together with roasted melon seed extract.

The compositions of the present invention may further comprise suitable carriers, excipients and diluents conventionally used in the manufacture of pharmaceutical compositions. In addition, it can be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, oral preparations such as syrups and aerosols, external preparations, suppositories and sterilized injection solutions according to a conventional method. Suitable formulations known in the art are preferably those as disclosed in Remington ' s Pharmaceutical Science, recently, Mack Publishing Company, Easton PA. Examples of carriers, excipients and diluents that can be included in the composition include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose , Microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. When the composition is formulated, it is prepared using a diluent such as a filler, an extender, a binder, a wetting agent, a disintegrant, a surfactant, or an excipient usually used. Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like, which may contain at least one excipient such as starch, calcium carbonate, sucrose, lactose, Gelatin and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral administration include suspensions, solutions, emulsions, syrups and the like. Various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included in addition to water and liquid paraffin, which are simple diluents commonly used. have. Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Examples of the non-aqueous solvent and suspension include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. Examples of the suppository base include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin and the like.

The term "administering" as used herein is meant to provide any desired composition of the invention to an individual by any suitable method.

The preferred dosage of the pharmaceutical composition of the present invention varies depending on the condition and the weight of the patient, the degree of disease, the drug form, the administration route and the period, and can be appropriately selected by those skilled in the art. For a desired effect, the composition of the present invention may be administered in an amount of 1 mg / kg to 1000 mg / kg per day. The composition may be administered once a day, or divided into several doses.

The pharmaceutical composition of the present invention may be administered to the individual by various routes. All modes of administration may be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intra-uterine dural or intracerebral injection.

The pharmaceutical composition of the present invention can be used alone or in combination with methods using surgery, radiation therapy, hormone therapy, chemotherapy, and biological response modifiers for the prevention or treatment of cancer.

In the present invention, the term "health functional food" refers to a food having a biological control function such as prevention and improvement of disease, bio-defense, immunity, recovery of disease and aging inhibition.

The roasted melon seed extract of the present invention can be added to health functional foods for the purpose of preventing or improving cancer. When the roasted melon seed extract of the present invention is used as a food additive, the roasted melon seed extract may be directly added or used together with other food or food ingredients, and may be appropriately used according to a conventional method. The amount of the active ingredient to be mixed can be suitably determined according to the intended use (prevention, health or therapeutic treatment). Generally, the roasted melon seed extract of the present invention is added in an amount of not more than 15% by weight, preferably not more than 10% by weight based on the raw material. However, in the case of long-term intake for the purpose of health and hygiene or for the purpose of controlling health, the amount may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount exceeding the above range.

There is no particular limitation on the kind of the food. Examples of the foods to which the above substances can be added include dairy products including meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen noodles, gums, ice cream, various soups, drinks, tea, Alcoholic beverages, and vitamin complexes, all of which include health foods in a conventional sense.

The health beverage composition of the present invention may contain various flavors or natural carbohydrates as an additional ingredient such as ordinary beverages. The natural carbohydrates may be monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, natural sweeteners such as dextrin and cyclodextrin, synthetic sweeteners such as saccharine and aspartame, and the like. The ratio of the natural carbohydrate is generally about 0.01 to 10 g, preferably about 0.01 to 0.1 g per 100 ml of the composition of the present invention.

In addition to the above, the composition of the present invention may further contain various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, A carbonating agent used in a carbonated beverage, and the like. In addition, the composition of the present invention may comprise flesh for the production of natural fruit juices, fruit juice drinks and vegetable drinks. These components may be used independently or in combination. Although the ratio of such additives is not critical, it is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.

Hereinafter, preferred embodiments, experimental examples, and production examples are provided to facilitate understanding of the present invention. However, the following examples, experimental examples and production examples are provided only for the purpose of easier understanding of the present invention, and the present invention is not limited by the examples, experimental examples and production examples.

Example  1. roasted melon seed extract and Fraction  Produce

The melon seeds used in the present invention are mixed seeds such as non-commodity discarded in Seongju-gun, and the representative cultivar, Obok honey, Melon seed, Chilo honey, Melon seed, etc. obtained from the fermentation section. In order to prepare the roasted melon seed extract, the melon seeds were first dried naturally and then roasted at 250 ° C for 120 minutes using a roasting machine. 10 g of each of the pulverized melon seeds was put into a round flask and degreased with a hexane organic solvent for 3 hours twice in a vertical extractor equipped with a reflux condenser to obtain roasted melon seed hexane extract. After the hexane degreasing process, the extract was continuously extracted three times with ethanolic organic solvent for 3 hours to obtain roasted melon seed ethanol extract. In addition, roasted melon seeds were extracted with distilled water three times three times to obtain roasted melon seed water extract. Each extract solution was filtered through Whatman No. 2 filter paper, and the filtrate was concentrated under reduced pressure at 45 占 폚 using a rotary vacuum concentrator.

Ethyl acetate (EtOAc) fractions and butanol (n-butanol) fractions were sequentially obtained from the roasted melon ethanol extracts using ethyl acetate and butanol in order to prepare roasted melon seed fractions.

Experimental Example  1. Measurement of anticancer activity of roasted melon seed extract

1-1. crystal  Violet Assay

In order to measure the anti-cancer activity of the fried melon seed extract prepared in Example 1, using a rat hepatoma cell line hepa1c1c7 a preliminary experiment Drysdale et al. 0.0 > (1986). ≪ / RTI > More specifically, Hepa1c1c7 cells were seeded into 96-well plates at a concentration of 1 × 10⁴ and cultured for 24 hours. The roasted melon seed extract was added to various concentrations (50, 100, 200 μg / ml) Lt; / RTI > After the incubation, the medium was removed and 0.2% crystal violet / 2% EtOH solution was added and stained for 10 minutes and then washed with distilled water. 0.5% SDS / 50% EtOH was added to each well and absorbance was measured at 610 nm. As a control group, roasted sesame seed extract was used. The results are shown in Fig.

As shown in Fig. 1, the hexane extract and the ethanol extract of the roasted melon seeds showed significantly higher anticancer activity than the roasted melon seed extract.

1-2. MTT Assay

To determine the anticancer activity of the roasted melon seed extract prepared in Example 1, MTT (3- (4,5-dimethylthiazol-2-yl) -2,5- diphenyltetrazolium bromide) assay was performed. (Human breast cancer cell line), AGS (human stomach cancer cell line, human gastric cancer cell line), HT-29 (human colon cancer cell line, human colorectal cancer cell line) , A549 (human lung cancer cell line) and HepG2 (human liver cancer cell line) were purchased from KCLB. All cancer cells were treated with 10% FBS (fetal bovine serum) And cultured in RPMI 1640 medium supplemented with 1% antibiotic (100 unit / ml penicillin, 100 μg / ml streptomycin) or DMEM medium in a 5% CO ₂ incubator at 37 ° C. Each of the cultured cancer cells was cultured for 24 hours at a concentration of 2 × 10 4 cells / ml in a 96-well plate. Then, roasted melon seed extracts (water extract, hexane extract and ethanol extract) 200 μg / ml) was added to each well and incubated for 48 hours in a 5% CO ₂ and 37 ° C incubator. After the culture, the medium was removed, MTT solution was added, and the culture was further cultured for 4 hours. The resulting formazan crystals were dissolved in DMSO (dimethylsulfoxide) and absorbance was measured at 540 nm. The relative cell proliferation inhibition rate was determined with the number of cells in the control group as 100%. The results are shown in Figs. 2 to 6.

As shown in FIG. 2 to FIG. 6, the ethanol extract of roasted melon seed extract showed the highest anticancer activity, and the water extract showed the lowest anticancer activity. In particular, roasted melon seed ethanol extracts showed excellent anticancer activity in liver cancer cell line (HepG2) and breast cancer cell line (MCF-7).

(MCF-7, A549, HT-29, HepG2, AGS) and mouse hepatocarcinoma cell line (human hepatocellular carcinoma cell line) in order to confirm the anticancer activity of the roasted melon seed ethanol extract which showed the highest anticancer activity through the above- (Hepa1c1c7), MTT assays were performed in the same manner. The results are shown in Fig.

As shown in FIG. 7, the roasted apple melon seed ethanol extract was 40% for Hepa1c1c7 cells, 62% for HepG2 cells, 63% for MCF-7 cells and 77% for A549 cells at a concentration of 200 μg / ml, Cell viability was 84% for HT-29 cells and 87% for AGS cells.

Experimental Example  2. roasted melon seeds Fraction  Measurement of antitumor activity

In order to measure the anticancer activity of the roasted apple melon seed fractions prepared in Example 1, roasted melon seed ethanol extract, roasted melon seed bean fraction and roasted melon seed ethyl acetate fraction were used to measure MTT The assay was performed. The results are shown in Fig. 8 and Fig.

As shown in FIG. 8 and FIG. 9, the non-polar fraction ethyl acetate fraction exhibited excellent anticancer activity in hepatocarcinoma cell line (HepG2) and breast cancer cell line (MCF-7) rather than the butanol fraction in polar fraction.

Hereinafter, a pharmaceutical composition containing the composition of the present invention and an example of a preparation for a health functional food will be described, but the present invention is not intended to be limited but is specifically described .

Formulation example  1. Preparation of pharmaceutical compositions

1-1. Sanje  Produce

Roasted melon seed extract 20 mg

Lactose 100 mg

Talc 10 mg

The above components are mixed and filled in airtight bags to prepare powders.

1-2. Manufacture of tablets

Roasted melon seed extract 10 mg

Corn starch 100 mg

Lactose 100 mg

Magnesium stearate 2 mg

After mixing the above components, tablets are prepared by tableting according to the usual preparation method of tablets.

1-3. Preparation of capsules

Roasted melon seed extract 10 mg

Crystalline cellulose 3 mg

Lactose 14.8 mg

Magnesium stearate 0.2 mg

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

1-4. Injection preparation

Roasted melon seed extract 10 mg

180 mg mannitol

Sterile sterilized water for injection 2974 mg

_{Na 2 HPO 4 · 2H 2 O} 26 mg

(2 ml) per 1 ampoule in accordance with the usual injection preparation method.

1-5. Liquid  Produce

Roasted melon seed extract 20 mg

10 g per isomer

5 g mannitol

Purified water quantity

Each component was added and dissolved in purified water according to the usual liquid preparation method, and the lemon flavor was added in an appropriate amount. Then, the above components were mixed, and purified water was added thereto. The whole was added with purified water to adjust the total volume to 100 ml, And sterilized to prepare a liquid preparation.

Formulation example  2. Preparation of food composition

2-1. Manufacture of health food

Roasted melon seed extract 100 mg

Vitamin mixture quantity

Vitamin A acetate 70 μg

Vitamin E 1.0 mg

Vitamin B1 0.13 mg

0.15 mg of vitamin B2

Vitamin B6 0.5 mg

Vitamin B12 0.2 μg

Vitamin C 10 mg

Biotin 10 μg

Nicotinic acid amide 1.7 mg

Folic acid 50 μg

Calcium pantothenate 0.5 mg

Mineral mixture quantity

1.75 mg of ferrous sulfate

0.82 mg of zinc oxide

Magnesium carbonate 25.3 mg

Potassium monophosphate 15 mg

Secondary calcium phosphate 55 mg

Potassium citrate 90 mg

Calcium carbonate 100 mg

Magnesium chloride 24.8 mg

Although the composition ratio of the above-mentioned vitamin and mineral mixture is comparatively mixed with a composition suitable for health food as a preferred embodiment, the compounding ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional method for producing healthy foods , Granules can be prepared and used in the manufacture of health food compositions according to conventional methods.

2-2. Health drink  Produce

Roasted melon seed extract 100 mg

Vitamin C 15 g

Vitamin E (powder) 100 g

19.75 g of ferrous lactate

3.5 g of zinc oxide

Nicotinic acid amide 3.5 g

Vitamin A 0.2 g

Vitamin B1 0.25 g

Vitamin B2 0.3g

Water quantification

The above components were mixed according to a conventional health drink manufacturing method, and the mixture was stirred and heated at 85 DEG C for about 1 hour. The solution thus prepared was filtered and sterilized in a sterilized 2 liter container, It is used in the production of the health beverage composition of the invention.

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.

Claims (7)

A pharmaceutical composition for preventing or treating cancer comprising roasted melon seed extract as an active ingredient.
[2] The roasted melon seed extract according to claim 1, wherein the roasted melon seed extract is extracted with at least one solvent selected from the group consisting of water, an alcohol having 1 to 4 carbon atoms, and a mixed solvent thereof. A pharmaceutical composition.
The pharmaceutical composition for preventing or treating cancer according to claim 2, wherein the alcohol having 1 to 4 carbon atoms is ethanol.
The roasted melon seed extract according to claim 1, wherein the roasted melon seeds are extracted with at least one solvent selected from the group consisting of water, an alcohol having 1 to 4 carbon atoms and a mixed solvent thereof, and then fractionated with ethyl acetate or butanol ≪ / RTI > or a pharmaceutically acceptable salt thereof.
The pharmaceutical composition for preventing or treating cancer according to claim 1, wherein the roasted melon seed extract is roasted at 100 to 300 ° C for 30 minutes to 10 hours.
The method of claim 1, wherein the cancer is selected from the group consisting of gastric cancer, colon cancer, breast cancer, lung cancer, liver cancer, non-small cell lung cancer, bone cancer, pancreatic cancer, skin cancer, head or neck cancer, melanoma in skin, Cancer, small intestine cancer, endometrioid cancer, thyroid cancer, adenocarcinoma, adenocarcinoma, adenocarcinoma, adenocarcinoma, small bowel cancer, adenocarcinoma, small bowel cancer, endometrioid adenocarcinoma, Neoplasms, renal cell carcinomas, renal pelvic carcinomas, hematologic cancers, brain cancers, central nervous system carcinomas, thyroid carcinomas, thyroid carcinomas, adenocarcinomas, soft tissue sarcomas, urethral cancers, penile cancer, prostate cancer, chronic leukemia, acute leukemia, lymphocytic lymphoma, , A primary central nervous system lymphoma, a spinal cord tumor, a brain stem glioma, and a pituitary adenoma.
Roasted melon seed extract as an active ingredient.

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