KR20200008272A - Composition for preventing or treating a cancer comprising cheonggukjang extract containing Sparassis crispa poder - Google Patents

Abstract

The present invention relates to a composition for preventing or treating cancer containing a fast-fermented bean paste extract which contains a Sparassis crispa powder. The fast-fermented bean paste extract containing the Sparassis crispa powder of the present invention not only has excellent DPPH radical scavenging ability and ABTS radical scavenging ability, but also has a high total flavonoid content and total polyphenol content, thereby having a high antioxidant effect. In addition, the fast-fermented bean paste extract has an effect of inhibiting cancer cell proliferation, thereby being able to be usefully used as a pharmaceutical composition for cancer treatment and health functional food or a health food composition for preventing or alleviating cancer.

Description

Composition for preventing or treating a cancer comprising cheonggukjang extract containing zinnia mushroom powder {Composition for preventing or treating a cancer comprising cheonggukjang extract containing Sparassis crispa poder}

The present invention relates to a composition for the prevention or treatment of cancer, including the Chungkukjang extract containing the mushroom mushroom powder.

A few mushrooms are found in coniferous forests over 1,000 meters above sea level and can be collected shorter from early summer to early autumn than other mushrooms. Blossom mushrooms are white or pale yellow, and bloom like white flowers or coral reefs. The growth period is considerably longer than other mushrooms. Adult mushrooms are found in small numbers in coniferous forests over 1,000 meters above sea level. It can be collected shortly until early autumn. Blossom mushrooms are white or pale yellow, and bloom like white flowers or coral reefs. The growth period is considerably longer than other mushrooms, and it takes four to six months to become adults. The main components of zinnia mushroom include beta glucan (β-glucan), chitosan, ergosterol, trehalose, etc., which shows the effects and effects of anticancer activity, diabetic disease improvement, and antioxidant.

Cheonggukjang is one of Korea's representative fermented foods made from soybean. It is one of the traditional types of soy sauce, soybean paste, red pepper paste, and soybean paste that have been commonly used to this day. The state has played an important role as a food. Cheonggukjang is made by using Bacillus subtilis called Bacillus subtilis attached to rice straw, which gives its unique taste and smell by enzymes produced by Bacillus subtilis during fermentation. In addition, it produces a large amount of viscous substance which is a mixture of levan form fructan and polyglutamate derived from sugar and protein of raw soybeans. It is a high-quality fermented soybean with high fat content.

Recently, many researchers have revealed the active ingredient contained in Cheonggukjang and its role, so isoflavone (isoflavone), phytic acid, saponin, trypsin derived from soybean It is a functional food because it contains a large amount of various biologically active substances such as inhibitors, unsaturated fatty acids, dietary fiber, oligosaccharides, and γ-glutamyltranspeptidase (γ-GTP), antioxidants and thrombolytic enzymes. Its importance is being reexamined.

In this regard, the present inventors confirmed that Cheonggukjang extract containing the mushroom mushroom powder not only has excellent DPPH radical scavenging ability and ABTS radical scavenging ability, but also has high antioxidant activity due to high total flavonoid content and total polyphenol content, and inhibits cancer cell proliferation. The present invention has been completed.

Korea Patent Registration No. 10-1712704

An object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of cancer, including the extract of Cheonggukjang containing the powder of Sparassis crispa .

Another object of the present invention is to provide a health functional food composition for preventing or improving cancer, including the extract of Cheonggukjang containing the powder of Sparassis crispa .

Another object of the present invention to provide a health food composition for the prevention or improvement of cancer, including the extract of Cheonggukjang containing the powder of Sparassis crispa .

It is another object of the present invention to provide a method for producing an extract of Cheonggukjang containing the powder of Sparassis crispa .

In order to achieve the above object,

The present invention provides a pharmaceutical composition for the prevention or treatment of cancer, including the extract of Cheonggukjang containing the flowers of Sparassis crispa .

In another aspect, the present invention provides a health functional food composition for preventing or improving cancer, including the extract of Cheonggukjang containing the flowers of Sparassis crispa .

Furthermore, the present invention provides a health food composition for preventing or improving cancer, including an extract of Cheonggukjang containing the powder of Sparassis crispa .

Furthermore, the present invention washes the white beans and then soaked in soybeans for 1-5 hours in water of 5-25 ℃, and then ripened for 4-8 hours at 80-150 ℃, and cooled to 30-60 ℃ (step 1 );

Fermenting the cooled soybeans and rice straw of step 1 at a temperature of 35-50 ° C. for 24-36 hours, drying the hot beans at a temperature of 40-80 ° C. for 4-8 hours, and then grinding to prepare Cheonggukjang powder (step 2). ); And

Mixing the chungkukjang powder and the mushroom mushroom powder and extracting them with an organic solvent (step 3);

It provides an extract manufacturing method of the Chungkukjang containing the mushroom mushroom ( Sparrassis crispa ) powder comprising a.

Cheonggukjang extract containing the mushroom mushroom powder of the present invention not only has excellent DPPH radical scavenging ability and ABTS radical scavenging ability, but also has high total flavonoid content and total polyphenol content, which has high antioxidant activity and inhibits cancer cell proliferation. It can be usefully used as a nutraceutical composition and nutraceutical or health food composition for the prevention or improvement of cancer.

Figure 1 is the result of measuring the total flavonoid content of the Chungkukjang extract containing the mushroom powder of the present invention.
Figure 2 is a result of measuring the total polyphenols of the Chungkukjang extract containing the mushroom mushroom powder of the present invention.
Figure 3 is a result of measuring the alpha, beta glucan content of the Chungkukjang extract containing the mushroom mushroom powder of the present invention.
Figure 4 is the result of measuring the DPPH radical scavenging activity of the Chungkukjang extract containing the mushroom mushroom powder of the present invention.
Figure 5 is the result of measuring the ABTS radical scavenging activity of the Chungkukjang extract containing the mushroom mushroom powder of the present invention.
Figure 6 is the result of measuring the SOD-like activity of the Chungkukjang extract containing the mushroom mushroom powder of the present invention.
Figure 7 is the result of measuring the anticancer activity of the extract of Chungkukjang containing the mushroom powder of the present invention.
Figure 8 is a photograph of the chungkukjang powder, chungkukjang hwan and chungkukjang flakes containing the mushroom mushroom powder prepared according to the standard manufacturing process of the present invention.

Hereinafter, the present invention will be described in detail.

Pharmaceutical composition for the prevention or treatment of cancer

The present invention provides a pharmaceutical composition for the prevention or treatment of cancer, including the extract of Cheonggukjang containing the flowers of Sparassis crispa .

In the present invention, the term "extract" can be prepared using common extraction methods, separation and purification methods known in the art. The extraction method is not limited thereto, but may be preferably a method such as hot water extraction, hot water extraction, cold needle extraction, reflux cooling extraction, or ultrasonic extraction.

In one embodiment of the present invention, the extract is characterized in that extracted with water, alcohol having 1 to 4 carbon atoms or a mixed solvent thereof, preferably 70% ethanol can be used.

In one embodiment of the present invention, the cancer is one selected from the group consisting of stomach cancer, colon cancer, breast cancer, uterine cancer, cervical cancer, ovarian cancer, lung cancer, liver cancer, small intestine cancer, esophageal cancer, kidney cancer, pancreatic cancer and bone marrow cancer Above, preferably gastric cancer.

In the present invention, 'treatment' refers to reversing, alleviating, inhibiting, or preventing the disease or condition to which the term applies, or one or more symptoms of the disease or condition, unless otherwise stated. As used herein, the term treatment refers to the act of treating when 'treating' is defined as above.

In one embodiment, the pharmaceutical composition is a powder, granules, tablets, capsules, suspensions, emulsions, syrups, oral formulations of aerosols, liquid, cream, lotion, paste or solid transdermal topical formulations, One or more formulations selected from the group comprising suppositories, sterile injectable solutions, and sprays.

The therapeutically effective amount of the composition of the present invention may vary depending on several factors, such as the method of administration, the site of interest, the condition of the patient, and the like. Therefore, when used in humans, the dosage should be determined in an appropriate amount in consideration of both safety and efficiency. It is also possible to estimate the amount used in humans from an effective amount determined through animal testing. Such considerations when determining the effective amount include, for example, Hardman and Limbird, eds., Goodman and Gilman's The Pharmacological Basis of Therapeutics, 10th ed. (2001), Pergamon Press; And E.W. Martin ed., Remington's Pharmaceutical Sciences, 18th ed. (1990), Mack Publishing Co.

Compositions of the present invention may also include carriers, diluents, excipients or combinations of two or more commonly used in biological agents. Pharmaceutically acceptable carriers are not particularly limited so long as the composition is suitable for in vivo delivery, for example, Merck Index, 13th ed., Merck & Co. Inc. Compounds, saline solution, sterile water, Ringer's solution, buffered saline solution, dextrose solution, maltodextrin solution, glycerol, ethanol and one or more of these components can be mixed and used as needed. Conventional additives may be added. In addition, diluents, dispersants, surfactants, binders and lubricants may be additionally added to formulate into main dosage forms, pills, capsules, granules or tablets such as aqueous solutions, suspensions, emulsions and the like. Furthermore, it may be preferably formulated according to each disease or component by a suitable method in the art or using a method disclosed in Remington's Pharmaceutical Science (Mack Publishing Company, Easton PA, 18th, 1990).

The composition of the present invention may further contain one or more active ingredients exhibiting the same or similar functions.

The pharmaceutical composition of the present invention may further include a pharmaceutically acceptable additive, wherein the pharmaceutically acceptable additive may include starch, gelatinized starch, microcrystalline cellulose, lactose, povidone, colloidal silicon dioxide, calcium hydrogen phosphate, Lactose, mannitol, syrup, gum arabic, pregelatinized starch, corn starch, powdered cellulose, hydroxypropyl cellulose, oppadry, sodium starch glycolate, carnauba lead, synthetic aluminum silicate, stearic acid, magnesium stearate, aluminum stearate, stearic acid Calcium, sucrose, dextrose, sorbitol, talc and the like can be used. The pharmaceutically acceptable additive according to the present invention is preferably included 0.1 parts by weight to 90 parts by weight based on the composition, but is not limited thereto.

The compositions of the present invention may be administered orally or by oral administration (eg, applied intravenously, transdermally, intraperitoneally, or topically) according to the desired method, and the dosage is based on the weight, age, sex, health status, The range varies depending on the diet, the time of administration, the method of administration, the rate of excretion and the severity of the disease.

Liquid preparations for oral administration of the compositions of the present invention include suspensions, solvents, emulsions, syrups, and the like, and various excipients such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. And the like may be included together. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized formulations, suppositories, and the like.

Nutraceutical or health food composition

The present invention provides a health functional food or health food composition for preventing or improving cancer, including the extract of Cheonggukjang containing the flower of Sparassis crispa .

When the extract of Cheonggukjang containing the mushroom mushroom powder of the present invention is used as a health functional food or a health food composition, the type of food is not particularly limited. Examples of foods to which the extract of Cheonggukjang containing the mushroom mushroom powder of the present invention may be added include drink, meat, sausage, bread, biscuit, rice cake, chocolate, candy, snacks, confectionary, pizza, ramen, other noodles, gum, Dairy products including ice cream, various soups, beverages, alcoholic beverages and vitamin complexes, dairy products, dairy products, capsules, pills, foods in the form of liquids or liquids, etc. Includes all of them.

The dietary supplement or health food composition containing the extract of Cheonggukjang containing the mushroom mushroom powder according to the present invention may be added as it is or used with other foods or food ingredients, and may be suitably used according to a conventional method. . The mixing amount of the extract of Chunggukjang containing the zinnia mushroom powder can be suitably determined according to the purpose of use (prevention or improvement). In general, the amount of the composition in the nutraceutical or health food composition may be added to 0.1 to 90 parts by weight of the total food weight. However, in the case of long-term intake for the purpose of maintaining health or for health control, the amount may be below the above range, and there is no problem in terms of safety, so the extract of Chunggukjang containing the zinnia mushroom powder is more than the above range. Can also be used as.

The health functional food or the health food composition of the present invention is not particularly limited to other ingredients except for containing the extract of Chunggukjang containing the mushroom mushroom powder of the present invention as an essential ingredient in the indicated ratio, and various flavors or Natural carbohydrates and the like may be included as additional components. Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose and the like; And conventional sugars such as polysaccharides such as dextrin, cyclodextrin, and sugar alcohols such as xylitol, sorbitol, erythritol. As flavoring agents other than those mentioned above, natural flavoring agents (tauumatin, stevia extract (for example, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. The ratio of the natural carbohydrate is generally about 1 to 20 g, preferably about 5 to 12 g per 100 nutraceutical or health food composition of the present invention.

In addition to the above, the health functional food or health food composition containing the extract of Cheonggukjang containing the mushroom powder of the present invention may be used in various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavoring agents, colorants, and the like. Neutralizers (cheese, chocolate, etc.), 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 drinks, and the like. Can be. In addition, the nutraceutical and health food composition of the present invention may contain the flesh for the production of natural fruit juice and fruit juice drinks and vegetable drinks.

These components can be used independently or in combination. The ratio of such additives is not so important but is generally selected in the range of 0.1 to about 20 parts by weight per 100 parts by weight of the dietary supplement or health food composition containing the extract of Cheonggukjang containing the mushroom mushroom powder of the present invention.

Blossom Mushroom ( Sparassis crispa ) Manufacturing Method of Extract of Cheonggukjang containing Powder

The present invention washes the white beans and then soaked in soybeans for 1-5 hours in water of 5-25 ℃, 4-8 hours at 80-150 ℃ ripening, cooling to 30-60 ℃ (step 1);

Fermenting the cooled soybeans and rice straw of step 1 at a temperature of 35-50 ° C. for 24-36 hours, drying the hot beans at a temperature of 40-80 ° C. for 4-8 hours, and then grinding to prepare Cheonggukjang powder (step 2). ); And

Mixing the chungkukjang powder and the mushroom mushroom powder and extracting them with an organic solvent (step 3);

It provides an extract manufacturing method of the Chungkukjang containing the mushroom mushroom ( Sparrassis crispa ) powder comprising a.

In the production method of the present invention, the organic solvent of step 3 is characterized in that the water, an alcohol having 1 to 4 carbon atoms or a mixed solvent thereof, preferably 70% ethanol.

In addition, in the production method of the present invention, characterized in that the mixture of the mushroom mushroom powder with respect to 100 parts by weight of the chungkukjang powder of step 3 to 2 to 30 parts by weight, preferably 100 parts by weight of the chungkukjang powder of step 3 Mushroom powder may be used by mixing in 20 to 25 parts by weight.

Hereinafter, the present invention will be described in more detail with reference to the following examples. However, the following examples are merely to illustrate the present invention, the contents of the present invention is not limited by the following examples.

Example 1 Preparation of Cheonggukjang Added with Blossom Mushroom Powder

High quality white beans (domestic) were selected and washed 5 times, immersed in water at 15 ° C. for 2hr, and then simmered at 100 ° C. for 6 hours and cooled to 50 ° C. Selected rice straw (pesticide-free dry rice straw by cultivating eco-friendly agricultural products, storage: fermentation chamber, shelf life: 1 year), fermentation (40 ℃, 48hr) and hot air drying (60 ℃, 6hr), and then powdered using a grinder (Particle size: 80 mesh).

1%, 2%, 5%, and 10% of the mushroom powder in powdered Cheonggukjang powder is mixed with purified water, respectively, and then subjected to primary molding (size mm) at a temperature of 16 ~ 20 ℃ using a depressor, followed by a pressing roller. Secondary molding was carried out, followed by hot air drying (60 ° C, 3hr) and contact drying (60 ° C, 2min), followed by drying, filling, and sealing to obtain an appropriate drying method based on the sensory test results.

Standardized processes for the development of Cheonggukjang powder, pill, and flake formulations have been developed, which are summarized in Tables 1, 2 and 3 below. Cheonggukjang powder produced according to the standard process was used as a raw material for the production of Cheonggukjang hwan and Cheonggukjang Flakes, the prepared Cheonggukjang Powder, Cheonggukjanghwan and Cheonggukjang Flakes are as shown in FIG.

Cheonggukjang Powder Process Name Process conditions Equipment name Raw material White Beans (Domestic) wash 5 times water washing Capital increase 100 ℃, 6hr Batch Fermentation 40 ℃, 48hr Fermentation chamber dry 60 ℃, 6hr dryer smash 80 mesh grinder

Cheonggukhwan Process Name Process conditions Equipment name Raw material Cheonggukjang Powder,
Blossom Mushroom Powder Mixed Cheonggukjang Powder: 47.2%
Blossom Mushroom Powder 10.0%,
Purified water 42.8% Molding (Refill) Ring Particles: Ψ 4.5 mm Refiller dry 60 ℃, 6hr (hot air drying) dryer coating No coating Filling, sealing capping Sealer

Cheonggukjang Flakes Process Name Process conditions Equipment name Raw material Cheonggukjang Powder,
Blossom Mushroom Powder Mixed Cheonggukjang Powder: 47.2%
Blossom Mushroom Powder 10.0%,
Purified water 42.8% Primary molding Ring Particles: Ψ 4.5 mm Refiller Secondary molding Flakes: Ψ 7.5 mm Noodle maker dry 60 ° C, 6hr (hot air drying) dryer coating No coating Filling, sealing capping Sealer

Experimental Example 1 Sensory Evaluation

For each of 10 trained sensory inspectors (who completed sample test tests in the Department of Food and Biotechnology, Faculty of Bio-Industrial Convergence), the intensity of Cheonggukjang's odor, texture, flavor, taste, and overall symbolicity was conducted on a nine-point scale. It was. The evaluation criteria for the intensity of Cheonggukjang are very weak (1 point)-weak (3 points)-normal (5 points)-strong (7 points)-very strong (9 points), and the texture, flavor, taste and overall preference The evaluation criteria for were very bad (1 point)-bad (3 points)-normal (5 points)-good (7 points)-very good (9 points).

 In order to determine the proper blending ratio of naturally fermented Chunggukjang powder and zinnia mushroom powder, the sensory characteristics of Cheonggukjang were mixed according to the mixing ratio.

The texture of texture, which is important for the formulation process development, was the lowest in the control and 1% of the mushroom mushroom mixing ratios of 4.1 and 4.2 points, and the highest in the 10% of the mushroom mushroom mixing ratio of 6.9 points. Flavor taste was highest in 6.1% of mushroom mushroom mixing ratio, but it was highest in 6.1. The lowest point was. The overall preference was highest in the cultivated mushroom mixture ratio of 10% with 6.6 points, and the lowest in the control group with mixed mushroom ratio of 0%. Overall, the preference of the flower mushroom mixture treatment was slightly higher than that of the control, and if the ratio of the flower mushroom mixture ratio was high, the odor smell of Cheonggukjang tended to be weakened by the mixture ratio of the flower mushroom, and the preference was low. The highest palatability of 10% was considered to be the optimal blending ratio between the functional hallucinations and flakes.

Sensory evaluation sample ¹)  Texture flavor Taste Overall
acceptability control 4.1 ± 1.4 5.0 ± 1.8 4.3 ± 1.3 4.1 ± 1.3 CGJAMP1 4.2 ± 1.5 5.2 ± 1.7 4.2 ± 1.5 4.2 ± 1.7 CGJAMP2 5.5 ± 1.6 5.3 ± 2.2 5.5 ± 1.6 5.4 ± 1.8 CGJAMP3 5.8 ± 1.4 5.6 ± 1.4 5.8 ± 1.5 6.1 ± 1.3 CGJAMP4 6.9 ± 1.2 6.1 ± 1.5 6.0 ± 1.3 6.6 ± 1.4

1) control: Ratio of cheonggukjang to Sparassis crispa powder is: 1: 0%. CGJAMP: cheonggukjang powder added with Sparassis crispa “s powder, CGJAMP1,2,3,4: ratios of Sparassis crispa powder is 1: 1%, 1: 2%, 1: 5%, 1: 10% respectively. n = 10, P <0.05.

Table 5 shows the results of sensory evaluation comparing the odor intensity of the control, natural fermented soybean soup (10% mixed with Chungkukjang powder and blossom mushroom powder), and conventional Chunggukjang.

sample ¹) Strength of order  Remark control 5.5 ± 1.1 CGJAMP4
(hot air drying method) 3.5 ± 1.3 Hot air drying method CGJAMP4
(contact drying method) 7.2 ± 2.4 Dry contact

1) control: Ratio of cheonggukjang to Sparassis crispa powder is: 1: 0%. CGJAMP: cheonggukjang powder added with Sparassis crispa powder, CGJAMP 4: ratios of Sparassis crispa powder is 1: 10%. n = 10, P <0.05

Odor of this evaluation was evaluated based on the odor degree (strength) peculiar to Cheonggukjang. The odor intensity of the control group was 5.5 points, which showed a moderate result. The hot air drying method (10% of the mixture of Cheonggukjang powder and flower mushroom powder) showed 3.5 points, which was lower than that of the control group. According to the contact drying method, the form of flakes prepared by the flower mixing ratio was 7.2 points, and the smell was stronger than the other two treatment groups. It was found that the scent of natural fermented soybeans mixed with the flower powder was weaker than the chungkukjang without the mixing of the flower powder, and the hot air drying method was more problematic than the contact drying in determining the heating method, which is the main drying process of the flake formulation. I think it is necessary to solve.

Example 2 Extract Preparation

Cheonggukjang powder containing the mushroom mushroom powder prepared in Example 1 was prepared using 70% ethanol. The 70% ethanol extract was extracted for 24 hours with stirring at room temperature with 10 times the solvent weight of the sample, and the same operation was repeated three times. The obtained extract was filtered using Whatman No. 1, concentrated using a rotary pressure reducer (N-1000, Eyela, Tokyo, Japan), and stored in a freezer at ­20 ° C. to be used as a sample.

Experimental Example 2 Measurement of Total Flavonoid Content

The total flavonoid content was sufficiently stirred in 4.3 mL of ethanol (95%), 0.1 mL of 10% aluminum chloride solution, and 0.1 mL of 1 M potassium acetate solution in 0.5 mL of extract according to Moreno et al., And left at room temperature for 40 minutes. Then absorbance was measured at 415 nm. The total flavonoid content of the extract was calculated using a standard calibration curve obtained in the range of 0-120 μg / mL with Quercetin as a standard, and the results are shown in FIG. 1. In the case of Cheonggukjang extract that did not contain the mushrooms, the total flavonoid content tended to increase proportionally as the concentration of the extract increased. In addition, the content of total flavonoids increased with increasing concentrations in the case of the extracts of the mushrooms which did not contain Cheonggukjang, and the difference between the total flavonoid contents of the Cheonggukjang extract and the mushroom extract was not significant. The total flavonoid content of the extracts containing 1%, 2%, 5%, and 10% of the mushrooms also showed little difference according to the content of the blossom mushrooms, and the total flavonoids contained in the mushrooms were known to be different. have.

Experimental Example 3 Measurement of Total Polyphenol Content

The phenol content of the sample was analyzed by measuring the absorbance at 760 nm using a spectrophotometer according to the Folin-Dennis method. Each sample was dissolved in DMSO (dimethylsulfoxide) at a certain concentration, 0.5 mL of each was taken in a test tube, and distilled water 7 mL was added, and 0.5 mL of Folin-Ciocalteau's phenol reagent was added and mixed for 1 minute. After adding 1 mL of saturated sodium carbonate solution, the mixture was allowed to stand at room temperature for 1 hour, and the absorbance was measured at 760 nm using a UV / VIS spectrophotometer (Optizen, Korea). To quantify the total phenolic content, tannic acid was dissolved in DMSO, prepared at a constant concentration, and tested in the same manner as the sample to prepare a calibration curve and to measure the total phenolic content of each extract.

The measured total phenolic content was found to increase in a concentration-dependent manner in both the extracts of Cheonggukjang, zinnia mushroom, and chungkukjang extract added with different concentrations of zinnia mushroom, as shown in Figure 2, 43.12 mg / g at 1000ppm concentration The total phenolic content of was measured, while the total phenolic content of 74.78 mg / g was measured in the zinnia mushroom, and it was found to contain approximately 1.7 times more than the chungkukjang. In addition, it was confirmed that the content increased rapidly in the Cheonggukjang extract added with 10% blossom mushroom reached 90% of the total phenolic content of the blossom mushroom. Based on the total phenolic content alone, it was concluded that adding 10% of the zinnia mushroom was most effective.

Experimental Example 4 Measurement of Alpha- and Beta-Glucan Content

Beta-glucan content was measured using a megazyme kit (mushroom and yeast β-glucan assay procedure kit). 1.5 mL of 37% HCl was added to 100 mg of the mushroom mushrooms, and stirred for 45 minutes in a 30 ° C. water bath. Then, 10 mL of tertiary distilled water was added and stirred for 2 hours in a 100 ° C. water bath. 10 mL of 2 N KOH was added to the reaction solution at room temperature, followed by adding 0.2 M sodium acetate buffer (pH 5.0). The sample obtained was centrifuged (1,500 g, 10 minutes) to obtain a supernatant, and 20 U / mL of external-1,3-betaglucan hydrolase (exo-1,3-β-glucanase) in 0.1 mL of the obtained supernatant. And beta glucosidase (β-glucosidase) 4 U / mL mixed solution was added 0.1 mL and reacted for 60 minutes in a 40 ℃ water bath. 3 mL of glucose oxidase / peroxidase (GOPOD) reagent was added to the reaction solution, and the total glucan content was obtained by measuring absorbance at 510 nm after 20 minutes of reaction.

Alpha-glucan content was added to 2 mL of 2 N KOH in 100 mg of the sample and stirred for 20 minutes in an ice-water bath. 8 mL of 1.2 M sodium acetate buffer (pH 3.8) was added to the reaction solution, and 1630 U / mL of amyloglucosidase and 0.2 mL of 500 U / mL of invertase were added thereto. After stirring for 30 minutes at centrifugation (1,500g, 10 minutes) to obtain a supernatant. 0.1 mL of 0.2M sodium acetate buffer (pH 5.0) and 3 mL of GOPOD reagent were added to 0.1 mL of the supernatant, and the absorbance was measured at 510 nm after 20 min reaction at 40 ° C. The absorbance of the total glucan and alpha glucan was calculated by g / 100 g value using the absorbance of the reaction solution in which the standard glucose solution was reacted with the GOPOD reagent. Beta glucan content was calculated by subtracting the alpha glucan content from the total glucan content.

As shown in FIG. 3, the beta glucan content of the Chunggukjang extract was measured to be 10.5g / 100g and the beta glucan content of the Zucchini mushroom extract was measured to be 18.7g / 100g. The beta glucan content of the added extract was approximately 11.5g / 100g, which was almost no difference compared to the Cheonggukjang, while the beta glucan content of the extract increased 106.0g / 100g to 16.0g / 100g. .

Based on these results, it is determined that the content of the blossom mushrooms added to Cheonggukjang is 10%.

Experimental Example 5 DPPH Radical Scavenging Activity

DPPH (α, α-diphenyl-β-picrylhydrazyl) radical scavenging activity was measured by modifying Blois et al. That is, 0.1 mL of 2 × 10 ^-4 M DPPH solution was added to 0.1 mL of each concentration of the extract, mixed for 10 seconds, and allowed to stand at room temperature for 60 minutes. DPPH radical scavenging ability was expressed as a percentage of the absorbance difference between the addition and the addition of the sample solution.

DPPH radical scavenging activity (%) = (1- (A _test- A _Blank / B _control ) × 100

A _test : absorbance of sample addition

A _Blank : Absorbance of Blank

B _control : Absorbance of no sample

The results of measuring antioxidant activity according to DPPH radical scavenging ability of the Chungkukjang, Blossom Mushroom, and Chungkukjang Extract containing Blossom Mushroom are summarized in FIG. 4. It was confirmed that DPPH radical scavenging ability increased in a concentration dependent manner as the concentration increased in all the samples examined. In the case of Cheonggukjang extract without the addition of zinnia mushroom, 9 ~ 16% of DPPH radical scavenging ability was confirmed, and in the case of the flower extract, 25 ~ 42% of DPPH radical scavenging activity was confirmed, which was more than double the DPPH radical scavenging ability of the extract. . In the case of extract added with zinnia mushroom in Chungkukjang, DPPH radical scavenging ability was increased with increasing the ratio of zinnia mushroom, and when 7% and 5% were added when 2% zinnia mushroom was added as compared to DPPH radical scavenging ability of 1000kgm extract When 32% and 10% were added, it was found that 81% DPPH radical scavenging activity increased, so that the synergistic effect of DPPH radical scavenging activity was the most effective addition of 10% of the mushroom mushrooms in Cheonggukjang.

Experimental Example 6 Measurement of Total Antioxidant Capacity by ABTS Cation Decolorization

Antioxidant activity of the samples by ABTS (2,2'-azino-bis-3-ethylbenzo-thiazoline-6-sulfonic acid) was measured according to Re et al. ABTS was dissolved in distilled water at a concentration of 7 mM, 2.4 mM potassium persulfate was added to generate ABTS radicals, and then used for 12-16 hours at room temperature in the dark. The radical generated ABTS solution was diluted with 99% ethanol and adjusted to have an absorbance of 0.70 ± 0.02 at 700 nm. The scavenging ability was measured by mixing 100 μL of the ABTS solution and 100 μL of the sample and reacting at 700 nm for 6 minutes.

ABTS radical scavenging acitivity (%) = (1- (A _test- A _Blank / B _control ) × 100

A _test : absorbance of sample addition

A _Blank : Absorbance of Blank

B _control : Absorbance of no sample

The results of measuring antioxidant activity according to the ABTS radical scavenging ability of the Cheonggukjang, Blossom Mushroom, and Cheonggukjang Extracts containing 1%, 2%, 5%, and 10% are as shown in FIG. 5. As with the DPPH radical scavenging results, it was confirmed that the concentration of ABTS radical scavenging activity increased with increasing concentration in all the samples examined. Cheonggukjang extract without the addition of zinnia mushroom showed 8 ~ 11% ABTS radical scavenging activity and 16 ~ 30% ABTS radical scavenging activity of the zinnia mushroom extract, more than twice the ABTS radical scavenging ability was confirmed. . In the case of the extract added with zinnia mushroom in the Chungkukjang, ABTS radical scavenging ability increased with the increase of the zinnia mushroom extract.In the case of Cheonggukjang with the addition of zinnia mushroom, 2% of the zinnia mushroom added in comparison with the ABTS radical scavenging ability The ABTS radical scavenging activity increased by 34% at the time of adding 5% of the mushroom mushrooms, and the ABTS radical scavenging ability increased by 104% when the mushroom mushrooms were added 5%, and the ABTS radical scavenging activity increased by 159% ABTS radical scavenging activity when the mushroom mushrooms were added at 10%. It was confirmed that the most effective effect of adding 5% of zinnia mushroom to Cheonggukjang at 1000ppm concentration.

Experimental Example 7 Measurement of Superoxide Dismutase-like Activity

SOD-like activity was measured according to Marklund's method. That is, 3 mL of tris-HCl buffer (50 mM tris [hydroxymethyl] amino-methane + 10 mM EDTA) and 0.2 mL of 7.2 mM pyrogarol were added to 0.2 mL of each sample and left at 25 ° C. for 10 minutes, followed by 1N HCl 1mL. The reaction was stopped, and the absorbance at 420 nm was measured for the amount of pyrogarol oxidized in the reaction solution. SOD-like activity expressed the difference in absorbance between the addition and no addition of the sample solution as a percentage.

SOD-like activity (%) = (1- (A _test- A _Blank / B _control ) × 100

A _test : Absorbance of sample adder

A _Blank : Absorbance of Blank

B _control : Absorbance of no sample

The results of measuring antioxidant activity according to SOD-like activity of the Cheonggukjang Extract, the Blossom Mushroom Extract, and the Cheonggukjang Extract containing 1%, 2%, 5% and 10%, respectively, are shown in FIG. 6. SOD-like activity measurement results As with ABTS and DPPH radical scavenging activity measurement results, it was confirmed that SOD-like activity increased in a concentration-dependent manner as the concentration increased in all the samples examined.

In the case of Cheonggukjang extract without the addition of zinnia mushroom, the SOD-like activity was found to be 5-8%, which was relatively low, whereas that of the zinnia mushroom extract was 22-35% SOD-like activity. Four times more SOD-like activity than the extract was confirmed. In the case of the extract added with zinnia mushroom, the SOD-like activity increased as the addition rate of zinnia mushroom increased.In the case of chungkukjang with the addition of zinnia mushroom in comparison with the SOD-like activity of the extract of cheonggukjang based on the highest concentration of 1000 ppm SOD-like activity increased by 87% when 2% of blossom mushrooms were added, SOD-like activity increased by 106% when 5% of blossom mushrooms were added, and SOD-like activity increased by 149% when 10% of blossom mushrooms were added. It was confirmed that the most effective synergistic effect was the addition of 2% of the mushroom mushrooms to Cheonggukjang at 1000ppm concentration.

Experimental Example 8 Anticancer Effect of Extract

The cancer cell line used in this experiment used human-derived gastric cancer cell AGS, and the normal cell line used HEK293, a human-derived kidney cell. Each cell was cultured in a 5% CO ₂ , 37 ° C incubator using RPMI 1640 medium supplemented with 10% fetal bovine serum (FBS), 100 U / mL penicillin G, and 100 μg / mL streptomycin. When higher, trypsin-EDTA was treated to passaging.

Toxicity to the cancer cells of the Chungkukjang extract containing the mushroom mushroom powder was confirmed by MTT assay according to the method of Ishiyamaem et al. Dispense 1 × 10 ⁵ cells / well into 96 well plates and incubate for 24 hours (37 ° C., 5% CO ₂ ), remove the medium used for pre-culture, and treat the medium with the respective sample concentration. Time incubation. After the incubation was completed, 10 μL of MTT solution (5 mg / mL) was added per well, followed by reaction for 4 hours at 37 ° C. and 5% CO ₂ incubator, followed by removing the medium containing the MTT reagent and adding 100 μL of DMSO. Color was developed at room temperature and the absorbance was measured at 550 nm using an ELISA reader.

MTT screening is a widely used assay along with sulforhodamin B (SRB) screening to detect cytotoxicity and cell proliferation. Living cells are reduced to a purple, non-soluble MTT formazan.

Cell survival rate (%) = (A _test / B _control ) × 100

A _test : absorbance of sample addition

B _control : Absorbance of no sample

Mutation plays an important role in the generation of cancer cells, and most carcinogens are known to have mutagenicity. Therefore, antimutagenic substances may have anti-cancer effects, and this effect was attempted to be verified in the extract of Cheonggukjang added with mushroom mushrooms. To this end, the effect of inhibiting the proliferation of human-derived gastric cancer cells A549 was measured using the extract of Cheonggukjang added with 2%, 5%, and 10% of the blossom mushrooms in Cheonggukjang, and the results are shown in FIG.

The proliferation inhibitory effect is judged to have a proliferation inhibitory effect when the proliferation of cancer cells inhibits the proliferation of cancer cells by more than 20% compared to the non-treatment group, and in the case of Cheonggukjang extract, the growth of gastric cancer cell line A549 at the concentration of 100-1000 ppm. It was confirmed that there is no inhibitory effect, but the inhibitory effect of growth of flower mushroom extract was not confirmed at low concentrations, but the effect of inhibiting the growth of gastric cancer cell line A549 was confirmed as the concentration of the flower mushroom extract increased. Cheonggukjang extracts containing 2% and 5% of zinnia mushrooms were found to have no effect of inhibiting the proliferation of gastric cancer cell line A549, as did the cheonggukjang extracts without zinnia mushrooms. In the case of the growth inhibitory activity of gastric cancer cell A549 was better than that of the flower mushroom extract, and it was confirmed that the cancer cell proliferation inhibitory activity increased with increasing concentration.

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 descriptive sense only and not for purposes of limitation. The scope of the invention is shown in the claims rather than the foregoing description, and all differences within the scope shall be construed as being included in the invention.

Claims (11)

A pharmaceutical composition for preventing or treating cancer, comprising extracts of Cheonggukjang containing the powder of Sparassis crispa .
The method of claim 1,
The extract is a composition, characterized in that extracted with water, alcohol having 1 to 4 carbon atoms or a mixed solvent thereof.
The method of claim 1,
The cancer is a pharmaceutical composition, characterized in that one or more selected from the group consisting of stomach cancer, colon cancer, breast cancer, uterine cancer, cervical cancer, ovarian cancer, lung cancer, liver cancer, small intestine cancer, esophageal cancer, kidney cancer, pancreatic cancer and bone marrow cancer.
The method of claim 3, wherein
The cancer is a pharmaceutical composition, characterized in that the stomach cancer.
Health functional food composition for the prevention or improvement of cancer, including the extract of Cheonggukjang containing the flowers of Sparassis crispa .
The method of claim 5,
The health functional food is a health functional food composition, characterized in that the food in the form of tablets, capsules, pills, flakes or liquids.
Health food composition for the prevention or improvement of cancer comprising an extract of Cheonggukjang containing the flowers of Sparassis crispa .
The method of claim 7, wherein
The health food is characterized in that it is selected from various drinks, flakes, meat, sausages, bread, candy, snacks, noodles, ice cream, dairy products, soups, ionic drinks, beverages, alcoholic beverages, gum, tea and vitamin complexes Composition.
Washing white beans and soaking the beans in water at 5-25 ° C. for 1-5 hours, then ripening at 80-150 ° C. for 4-8 hours and cooling to 30-60 ° C. (step 1);
Fermenting the cooled soybeans and rice straw of step 1 at a temperature of 35-50 ° C. for 24-36 hours, and drying the hot air for 4-8 hours at a temperature of 40-80 ° C., followed by grinding to prepare Cheonggukjang powder (step 2 ); And
Mixing the chungkukjang powder and the mushroom mushroom powder and extracting them with an organic solvent (step 3);
Extraction method of Chungkukjang containing the zinnia mushroom ( Sparrassis crispa ) powder comprising a.
The method of claim 9,
The organic solvent of step 3 is water, alcohol having 1 to 4 carbon atoms or a mixed solvent thereof.
The method of claim 9,
Method of producing a mixture of 2 to 30 parts by weight of the mushroom mushroom powder compared to 100 parts by weight of the cheongukjang powder of step 3.

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