KR20160033917A - A manufacturing method of high anti-cancer effect composition by continuous extraction of Phellinus and wood-cultivated ginseng, and the composition - Google Patents

Abstract

The present invention relates to a method for producing an anticancer effect enhancement composition by continuously and sequentially extracting Phellinus linteus and wood-cultivated ginseng. According to the method for producing an anticancer effect enhancement composition, firstly, Phellinus linteus is extracted by using hot water; continuously, wood-cultivated ginseng is additionally added to the Phellinus linteus and is extracted by using hot water; and secondly, Phellinus linteus and the wood-cultivated ginseng are extracted by using 50% alcohol. The purpose of the present invention is to provide a method for obtaining a composition with excellent anticancer activity from Phellinus linteus and a wood-cultivated ginseng extract.

Description

Technical Field [0001] The present invention relates to a method for preparing an anticancer composition by sequential sequential extraction of mud mushroom and goat ginseng,

The present invention relates to a method for producing an anticancer enhancing composition by sequential sequential extraction of mud mushroom and goat gum, and a composition thereof.

The inventors of the present invention found that when the active ingredient is obtained through first hot water extraction of pulverized mud mushroom, followed by hot water extraction with addition of dried ground ginseng, and then extraction with the second alcohol, the anticancer activity is enhanced I confirmed the fact newly.

In addition, we confirmed that the above active ingredient enhances the anticancer effect of existing anticancer drugs.

Furthermore, it was also confirmed that the anticancer effect is further enhanced when the active ingredient of the present invention contains other auxiliary ingredients such as Hwasungcho, Hwanggi, Saururus chinensis, Radix, Kale,

It has long been known that mushrooms are effective in the prevention and treatment of anticancer activity, antioxidant activity, cholesterol lowering effect, hypotensive action, immune strengthening action, tumor suppression, uterine bleeding and menstrual irregularity, have.

Among these mushrooms, the extract of mud mushrooms is loved as a health supplement. Mud mushroom is a white rot fungus belonging to Aphylophorales, Hymenochaetaceae, and Phellinus. Fruiting bodies are known as corky plants, either single-year or perennial. It grows in the form of a clone and grows in a hemispherical shape mainly attached to trees. Normally, they grow vertically and grow from 5cm to less than 14cm in size. The surface of the upper layer is dark brown or yellowish brown with villo and the surface of the hole is dark brown with 6 ~ 8 per mm. Spores are 4-5 x 3 ~ 3.5 ㎛ in elliptical shape and the surface is known to be smooth (Ryvardan, 1993). It is largely parasitic on hardwoods such as oak and mulberry. In the case of shiitake mushroom, it occurs in oak alley and causes disease (Cha et al., 1994).

The goat ginseng belongs to the jelly family and refers to the perennial ginseng germinated in the wild. 2004, 19 (4): 45-50). The genetic identification of Korean wild ginseng and American wild gibseng by using pyrosequencing method, Kor J. Herbology 2004; 19 (4): 45-50) 2004). In this study, we investigated the effects of anti-hyperglycemia and obesity on ICR mice, Arch Pharm Res., 2004; 27 (7): 790-796.) Studied the prevention of obesity.

On the other hand, according to the extraction method, the hot-water extract of muddy mushroom fruit body is known to exert an excellent effect for the prevention and treatment of the anticancer activity and acute respiratory diseases. However, in many cases, mud mushrooms are often used with other herbal extracts.

Accordingly, the present inventors have selected a medicinal plant which causes a synergistic effect when used in combination with a commonly used medicinal plant extract. As a result, it was confirmed that the goat ginseng extract had a synergistic effect with the mud mushroom extract.

It is an object of the present invention to provide an extraction method for obtaining a composition having excellent anticancer activity from mud mushroom and cane ginseng extract.

At the same time, it proves that mud mushroom and goat ginseng are synergistic according to the extraction method.

In order to accomplish the above object, the present invention provides a method for producing a cancer-

The mud mushroom is firstly extracted with hot water, and subsequently the goat's ginseng is further added thereto, followed by hot water extraction, followed by extraction with addition of 50% alcohol.

In this case, in the first continuous hot water extraction, the input ratio of mud mushroom and goat ginseng is 25-75%: 25-75% by weight.

The present invention is also an anticancer enhancing composition for enhancing anticancer activity of an existing anticancer drug by containing 10 to 90% of the above composition.

Also, the composition is characterized in that it is an anticancer enhancing composition comprising at least one of extracts of Hwasungcho, Hwanggi, Saururus chinensis, Radix kale, Kale, Ganoderma lucidum, Eugenia hippocampus, Drosophila melanogaster and Aspergillus oryzae as an auxiliary component.

More specifically, the auxiliary component is characterized in that it is at least one of an extract of Acanthopanax senticosus and Snow Flower Cordyceps.

The mud mushroom extract and the goat ginseng extract according to the present invention are not toxic and have an action to enhance the anticancer activity of the mud mushroom extract through the oxidation activity of the goat's ginseng extract. In particular, it has an excellent anticancer synergistic effect when it is used in combination with existing anticancer drugs, and thus can be usefully used as an anticancer composition.

1 is a diagram illustrating an extraction method according to the present invention.

The present invention provides an anticancer agent elevated composition comprising mud mushroom extract and goat ginseng extract as active ingredients.

The extracts of mud mushroom and goat ginseng, which are effective ingredients in the composition of the present invention, can be obtained by the following method.

First, the mud mushroom is washed with water to remove foreign matter, sediment and dried. Mud mushroom and goat ginseng can be cultivated or marketed without restriction. The dried mud mushroom and the goat ginseng are pulverized, and an appropriate amount of solvent is added to complete the immersion. The extraction method can be impregnated or warmed at room temperature.

As the extraction solvent, water and an alcohol having 1 to 4 carbon atoms can be used.

The extraction example will be described with reference to FIG. 1. First, the mud mushroom is pulverized with a primary extraction, water is added at a ratio of 1:20 (100 g of mud mushroom, 2 L of distilled water), and then primary extraction is performed at 100 ° C. for 4 hours. Then, 100 g of goat ginseng and 20 L of distilled water are added, followed by addition of hot water for 2 hours. After extracting hot water of mud mushroom and goat ginseng, add 20 L of 50% alcohol and perform the same addition for 4 hours. The extract is filtered, purified, concentrated and lyophilized to obtain a final component (hereinafter also referred to as 'PG').

The composition of the present invention can be made in a liquid form and a solid form.

The antioxidant activity or anticancer activity of the composition of the present invention can be measured by the P388 cell line as a standard cell line and the sarcoma cell line as a solid cancer cell line.

Hereinafter, preferred embodiments of the present invention will be described in order to facilitate understanding of the present invention. However, the following examples are provided only for the purpose of easier understanding of the present invention, and the present invention is not limited by the examples.

Example 1. Preparation of mud mushroom extract and goat ginseng extract

As shown in Fig. 1, continuous extraction was carried out by continuous sequential extraction (primary mud mushroom hydrothermal extraction and goat ginseng hydrothermal extraction extraction, secondary alcohol extraction by 50% perfusion extraction), followed by drying to obtain lyophilized powders.

The first extraction was performed by cutting the dried mud mushroom into small pieces, adding 2 L of distilled water to 100 g of the coarse ground product pulverized with a pulverizer (1:20), stirring at 100 ° C for 4 hours, Were ground to a diameter of less than 1 centimeter and added to the above dried mud mushroom extract and subjected to hot water extraction for 2 hours.

Secondary extraction was performed by adding 50% alcohol as a raw material at a ratio of 1 (ginseng g) to 20 (50% ginseng).

The extract obtained from the above process was filtered with filter paper (Whatman # 2), and the filtrate was lyophilized at -80 ° C to obtain a hot-water extract (PG) in the form of a dry powder.

As a result of comparing the anticancer activities of the PG prepared by the new extraction method and the dried mud mushroom extract and the goat ginseng extract by the conventional method at the same concentration, the PG extract obtained by the sequential sequential extraction method had much better anticancer effect than the extract obtained by the conventional extraction method (Fig. 1). Thus, the utility of the extraction method of the present invention can be confirmed.

Fig. 1. Anticancer properties of single sequential extract (PG) and single component extracts extracted by general extraction method

For the completion of the present invention, PG extracts with different amounts of dried mud mushroom and goat ginseng were obtained and the following tests were conducted.

Example 2 Preparation of Optimal Anticancer Activity Extract According to the Mixing Ratio of Dried Mud Mushroom and Goat's Ginseng

In order to complete the continuous extraction method of the new PG extract having excellent anticancer activity, the weight ratio of dried mud mushroom to goat ginseng was varied in consecutive order according to Table 1 (Table 1).

The sarcoma 180 used in the present invention was a mouse-derived tumor cell line and was distributed at the Korean Cell Line Bank.

Medium for culturing and maintenance of tumor cell lines was used RPMI 1640 with fetal bovine serum (FBS) supplemented with 10%, and penicillin and streptomycin the tissue culture flask 25cm ^2, each 10,000 units / ml and 10 mg / ml mixed culture And cultured in a 5% CO ₂ cell incubator at 37 ° C.

The anticancer activity was determined by the SRB method. The degree of cell growth of PLE, PBE, PGE and positive control DOX in two mouse-derived tumor cells was slightly modified by sulforhodamine (SRB) B (Kim et al., 1996) and microtetrazolium (MTT) assay (Mosmann et al., 1983) Respectively. First, SRB was inoculated on a 96-well plate so that the number of tumor cells was 10,000 per well. Then, the cells were cultured for 24 hours, and each test substance was inoculated and further cultured for 48 hours. 50% trichloroacetic acid (TCA) was added to each tumor cell for 4 hours and 2 hours, and the fixative was removed by rinsing with distilled water 5 times. Tumor cells were stained with 0.4% sulforhodamine B (SIGMA) for 30 min at room temperature, and Tris base (10 mM, pH 10.5) was added to dissolve stained dyes only in surviving tumor cells. MTT method was inoculated so that the number of cells per well was 5,000 and each test substance was inoculated before culturing differently from SRB method. Cultivation was carried out for 4 days after inoculation. After the incubation was completed, 50 ml of 3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide (MTT, SIGMA, USA) solution was added to each well and further cultured for 4 hours. After that, the medium was carefully removed from each well. Next, 150 ml of DMSO (dimethylsulphoxide) was added to dissolve the formazan crystal. Cell plates stained with SRB and MTT were measured for absorbance at 490 nm using a microplate reader (VERSAmax ™, Molecular Devices, USA). In order to evaluate the effect of the test substance on the tumor cells, the cell count was expressed in% by dividing the absorbance of the well treated with the test substance by the absorbance of the negative control well without treatment. Statistical analysis was performed using the SAS statistical package (release 8.1 SAS Institute Inc., Cary, North Carolina, USA) and the significance level was p <0.05.

As a result, Fig. 2 shows excellent effects in the case of the blending ratio of 2 to 4 as shown in Fig.

Fig. 2. Comparative study of anticancer activity according to the ratio of successive sequential extraction of dried mud mushroom and goat ginseng

Example 3: Combination ratio 4 and anticancer activity when the conventional anticancer drug alone was administered

The anticancer activity of Doxorubicin was confirmed first. The non - treatment group was based on 100% survival rate of cancer cells that were not treated with anticancer drugs. Fig. As shown in Fig. 3, the antitumor activity was increased when the concentration of doxorubicin was increased.

Fig. 3. Antitumor activity of doxorubicin

Next, the anticancer activity of the compounding ratio 4 was confirmed. The non - treatment group was based on 100% survival rate of cancer cells that were not treated with anticancer drugs. Fig. As shown in Fig. 4, the anticancer activity was increased when the dose was increased.

Fig. 4. Anticancer activity of compounding ratio 4

Example 4. Comparison of anticancer activity when the combination ratio of 4 and doxorubicin was concomitantly administered

Experiments were carried out with a mixing ratio of 4 as PG and doxorubicin as Dox.

Fig. 5. Combination ratio 4 and doxorubicin combined treatment

Dox 1/4 cancer cell survival rate was 95% and PG1 / 2 cancer cell survival rate was 73%. PG 1/2 + Dox 1/4 showed cancer cell survival rate of 56%. When Dox was fixed to 1/4 and PG was increased to 1/8, 1/4, and 1/2, the anti-cancer effect was increased. Therefore, PG may enhance the anticancer effect and reduce the use of anticancer drugs. As a result, side effects of anticancer drugs can be reduced. That is, the composition of the present invention can be used as an adjunct therapy.

Example 5. Comparison of antitumor activity between PG and adjuvant

The compounding ratio 4 is called PG (main component), and the auxiliary component extract (15% of sperm 5%, 3% of spermatozoa, 6% of spermatozoa, 1% of mulberry, 1% of kale, 2% ). (The auxiliary components were extracted in the same ratio as above and subjected to hot water extraction in the same manner as the PG extraction method.)

Fig. 6. When the compounding ratio 4 and the auxiliary component are used alone or in combination,

As a result, Fig. 6, PG and adjuvant alone showed cancer cell survival rates of 65% and 80%, respectively. However, when the main component and the auxiliary component were coadministered at a ratio of 1: 1 or 3: 1, it was confirmed that there was a synergistic effect or synergistic effect on the anticancer effect.

The anti-cancer effect was confirmed in Example 6 by administering the combination of PG and the individual extracts known to have anticancer activity in the auxiliary component extract.

Example 6. Comparison of antitumor activity between PG and adjunct ingredient extracts

The compounding ratio of 4 was designated as PG, and the anticancer activity of each of the adjunct ingredients was reported in combination with each of the components reported in the anticancer activity (Hwasungcho, Seomyoji, Cordyceps, Hansuo).

Fig. 7. Combination ratio 4 and antidepressant activity

Fig. As shown in Fig. 7, the anticancer activity of the combination with PG was superior to that of PG alone and PG when used alone.

In conclusion, the present invention demonstrates the excellent anticancer effect when PG is administered in combination with the above-described auxiliary component extract.

Taken together, the extract of the present invention is superior in anticancer activity to the single extract of each of dried mud mushroom and goat ginseng.

In addition, the extract of the present invention promotes the effects of existing anticancer drugs and has a better effect when it is combined with an extract of ancillary components such as a Chinese cabbage flower and a lily of the valley.

Therefore, the composition of the present invention can be effectively used for prevention and treatment of immune enhancement, anticancer drug supplements, anticancer drug combination administration.

Examples of formulations for the composition of the present invention are illustrated below.

Formulation Example 1: Preparation of pharmaceutical preparations

1. Manufacturing of powder

Continuous sequential extract of mud mushroom and goat juice 2g

Lactose 1g

The above components were mixed and packed in airtight bags to prepare powders.

2. Preparation of tablets

100 mg of mud mushroom and goat's sequential sequential extract

100 mg of corn starch

Lactose 100 mg

2 mg of magnesium stearate

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

3. Preparation of capsules

100 mg of mud mushroom and goat's sequential sequential extract

100 mg of corn starch

Lactose 100 mg

2 mg of magnesium stearate

After mixing the above components, the capsules were filled in gelatin capsules according to the conventional preparation method of capsules.

Formulation Example 2: Preparation of food

Foods comprising the mud mushroom and the sequential sequential extract of goat gum of the present invention were prepared as follows.

1. Preparation of cooking seasoning

Healthy cooking sauce was prepared from 20 to 95% by weight of the successive sequential extracts of mud mushroom and goat gum of the present invention.

2. Manufacture of tomato ketchup and sauce

0.2-1.0 wt% of the successive successive extracts of mud mushroom and goat mushroom of the present invention were added to tomato ketchup or sauce to make a tomato ketchup or sauce for health promotion.

3. Manufacture of flour food

0.5 to 5.0% by weight of mud mushroom and goat mushroom sequential extract of the present invention were added to wheat flour, and breads, cakes, cookies, crackers and noodles were prepared by using this mixture to prepare foods for health promotion.

4. Manufacture of soups and gravies

0.1 to 5.0% by weight of the successive successive extracts of mud mushroom and goat mushroom of the present invention were added to soups and gravies to prepare health promotion meat products, noodle soups and juices.

5. Manufacture of ground beef

A ground beef for health promotion was prepared by adding 10 wt% of the mud mushroom and goat's continuous sequential extract of the present invention to ground beef.

6. Manufacture of dairy products

5 to 10% by weight of the successive sequential extracts of mud mushroom and goat mushroom of the present invention were added to milk and various dairy products such as butter and ice cream were prepared using the milk.

7. Manufacturing of wires

Brown rice, barley, glutinous rice, and yulmu were dried by a known method and dried, and the mixture was granulated to a powder having a particle size of 60 mesh.

Black soybeans, black sesame seeds, and perilla seeds were steamed and dried by a conventional method, and then they were prepared into powder having a particle size of 60 mesh by a pulverizer.

The dried product was dried and pulverized with a granulator to a particle size of 60 mesh to obtain a dried powder.

The grains, seeds, and dried powder of mushroom and goat gum sequential extracts prepared above were blended in the following proportions.

Cereals (30% by weight of brown rice, 15% by weight of yulmu, 20% by weight of barley)

Seeds (7% by weight perilla, 8% by weight black bean, 7% by weight black sesame)

Dried powder (3 wt%) of mud mushroom and cane ginseng extract,

(0.5% by weight),

(0.5% by weight)

Formulation Example 3: Preparation of beverage

1. Manufacture of carbonated beverages

A syrup is prepared by mixing additives of 5 to 10% of sugar, 0.05 to 0.3% of citric acid, 0.005 to 0.02% of caramel and 0.1 to 1% of vitamin C and mixing 79 to 94% of purified water thereto, ° C for 20 to 180 seconds, mixed with cooling water at a ratio of 1: 4, and then 0.5 to 0.82% of carbon dioxide gas was injected to prepare carbonated beverages containing the mud mushroom and goat's continuous sequential extract of the present invention.

2. Manufacture of health drinks

Sub ingredient such as liquid fructose (0.5%), oligosaccharide (2%), sugar (2%), salt (0.5%) and water (75%) and mulberry mushroom and successive sequential extract of goat mushroom were uniformly blended And then packed in small containers such as glass bottles and PET bottles to produce health drinks.

3. Manufacture of vegetable juice

Healthy vegetable juice was prepared by adding 5 g of the sequential mud mushroom and the goat gum of the present invention to 1,000 ml of tomato or carrot juice.

4. Manufacture of fruit juice

Healthy fruit juice was prepared by adding 1 g of the sequential mud mushroom and goat gum sequential extract of the present invention to 1,000 ml of apple or grape juice.

Claims (5)

The mud mushroom is firstly extracted with hot water, the goat's ginseng is further added thereto, followed by hot water extraction, and then the second extract is added with 50% alcohol.
Methods for the preparation of anticancer enhancing compositions by sequential sequential extraction of mud mushroom and goat ginseng. The method according to claim 1,
Characterized in that, in the first continuous hot water extraction, the input ratio of mud mushroom and goat ginseng is 25 ~ 75%: 25 ~ 75%
Methods for the preparation of anticancer enhancing compositions by sequential sequential extraction of mud mushroom and goat ginseng. An anticancer composition for enhancing anticancer activity of an existing anticancer drug by containing 10 to 90% of the composition according to claim 1 or 2 to an existing anticancer drug. An anticancer composition according to any one of claims 1 to 3, wherein the composition comprises at least one of the extracts of Hwasungcho, Hwanggi, Saururus, Radix, Kale, 5. The anticancer composition according to claim 4, wherein the auxiliary component is at least one selected from the group consisting of extracts of Acanthopanax senticosus and Snow Flower Cordyceps.

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