KR20010000326A - Use for antitumor agent of extract of Phellinus linteus - Google Patents

Abstract

PURPOSE: A Phellinus linteus extract as an anti-cancer drug is provided, which does not show cytotoxicity in vitro cancer cells, but activates immunity of living body in vivo, so elongates survival rate and survival period of a mouse with ascites cancer and solid cancer, and shows suppressing effect on growth of solid cancer. CONSTITUTION: A process for the preparation of Phellinus linteus extract as anti-cancer drug comprises: adding 5 times of water to artificially raised Phellinus linteus reagent, and extracting at 80°C for 6hrs; filtering and extracting the residue again; adding the extracts, processing dialysis, freeze-drying to get the powder of water-extract fraction; adding cold acetone to the water-extract fraction till to be 50%, centrifuging the precipitates, and freeze-drying to get the powder of organic solvent precipitates fraction; examining cytotoxicity of artificially raised Phellinus linteus fruiting body in vitro(in result, the water-extract fraction or the organic solvent precipitates fraction of Phellinus linteus does not show cytotoxicity to cancer cells); examining in vivo anti-cancer activity of Phellinus linteus extract.

Description

Use of antitumor agent of extract of Phellinus linteus

The present invention relates to the use of gold dust mushroom Phellinus linteus as an anticancer agent. More specifically, the present invention relates to the effect of inhibiting cancer cells by enhancing the immunity of the water extract fraction or organic solvent precipitation fraction of artificially cultured gold dust mushroom fruiting body.

Polysaccharides with anticancer activity continue to be separated from natural sources such as higher plants, fungi, yeast, and bacteria (Takeda, T., Shibata, S. and Fukuoka, Chem. Pharm. Bull. (1969)). 17: 1910-1916; Kao, I, Kobayashi, S., Yokokura, T. and Mutai, M., Gann (1981) 72: 517-523). Among these polysaccharides, anticancer activity of basidiomycetes is the most widely studied (Chihara, G., Hanuram, J., Maeda, Y., Arai, Y. and Fukuoka, F., Nature (1970), 225: 943). (944), Chihara, et al., Are the high-density polymer β-1,3-glucan β-1,3-glucan, Lentinan (β-1,3-glucan), from the fruiting body of Lenitnus edodes. lentinan) (Chihara, G., Hanuram, J., Maeda, Y., Arai, Y., and Fukuoka, F., Nature (1969), 222: 687-678). Studies of the anticancer components of these basidiomycetes on immunity demonstrated that lentinan acts as an immumoaccelator for cellular immunity and as a T-cell adjuvant (Sasaki). , T. and Takasuka, N., Carbohyd.Res. (1976), 47: 99-104).

Meanwhile, studies on the anticancer component of Korean basidiomycetes revealed that hot water extracts of fruiting bodies such as cloud mushrooms, shiitake mushrooms, and oyster mushrooms had a strong blocking ability on sarcoma 180 (Kim, BK, Park). , EK and Shim, MJ, Sing. Arch. Pharm.Res. (1979), 2: 145-149. Kim et al. Reported that situational mushrooms have anticancer activity (Kim, HM, Han, SB, Oh). , GT, Kim, YH, Hong, DH and Yoo, LD, Int. J. Immunopharmacology (1996), 18: 295-303; Han SB, Lee, CW, Jeon, YJ, Hong, ND, Yoo, ID, Yang , KH and Kim, HM, Immunopharmacology (199 9), 41: 157-164). However, no studies on anticancer activity of artificially grown S. mushroom fruiting bodies have been reported.

In view of the above, the present inventors obtained the water extract fraction and the organic solvent precipitate fraction from the artificially-grown geumse jinja mushroom fruit body, and then added it to the cancer cell line to investigate the cytotoxicity in vitro, and sacoma in the mouse ( sarcoma) 180 was implanted to induce ascites and solid cancer, and then water extract fraction or organic solvent precipitation fraction of Geumsaeng mushroom mushroom fruit body was administered to investigate anti-cancer activity in vivo. The present invention was completed by investigating acute toxicity by administration.

Accordingly, an object of the present invention is to provide a cancer prevention and treatment composition comprising the situation mushroom fruiting body having an anticancer activity against cancer cells of the living body as an active ingredient.

Another object of the present invention is to provide a method for extracting the gold dust mushroom fruiting body.

The object of the present invention is to obtain the water extract fraction and the organic solvent precipitate fraction of artificially cultivated gold dust situation (Phellinus linteus) and add it to cancer cell lines SNU-C4, A-549, P-388, L-1210 To investigate the cytotoxicity in vitro, transplanted sarcoma 180 into mice to induce ascites cancer and solid cancer, and then, extracts from the fruiting body of Geumsaeng mushroom were administered to measure viability, prolonged survival effect and cancer cell weight. It has been confirmed that it has anti-cancer effect by enhancing biological immunity in vivo, and then subjected to acute toxicity test by oral administration to Geumsa Sacral Mushroom Extract SD rat, followed by general symptoms, observation of dead animals, weight change, feed intake, negative water change and autopsy It achieved by confirming the stability.

Hereinafter, the configuration of the present invention will be described.

The present invention is added to the fruiting body of artificial cultured Phellinus linteus (Phellinus linteus) after extracting for 6 hours at 80 ℃ and filtered and then dialyzed and lyophilized to obtain a water extraction fraction and the acetone 50 in the water extraction fraction Preparing an organic solvent precipitate by centrifuging and lyophilizing the precipitate by adding the precipitate so as to obtain a%; Adding artificial cultured gold dust mushroom extract and organic solvent precipitation fractions after culturing cancer cell lines SNU-C4, A-549, P-388, L-1210 and examining cytotoxicity against normal cells and cancer cells; To investigate the in vivo anticancer activity of Phellinus linteus extract of artificially cultivated Sacoma (sarcoma 180), inducing ascites cancer and solid cancer in mice, and then administering the extract of S. aureus and survival rate And measuring the cancer cell weight; Oral administration of Geumsa Sacral Mushroom Extract to SD rats followed by acute toxicity test by measuring mortality, weight change, clinical symptoms, feed intake and drinking volume.

In this step, Phellinus linteus (Phellinus linteus) was used to purchase the fruiting body of artificially cultivated plastic pot of the situation mushroom fruiting body in the Jinsa Situation mushroom farm. 3- (4,5-dimethylthiazole-2-yl) -2,5-diphenyltetrazolbromide (MTT, 3- (4,5-Dimethylthiazol-2-yl) -2,5-diphenyltetrazoliumbromide), trypsin (trypsin), RPMI1640 medium was purchased from Sigma, antibiotics-antimycotics (antibiotics-antimycotics), FBS was purchased from Difco. SNU-C4, A-549, P-388, L-1210 and MA104 cell lines were used by the Seoul National University Cell Line Bank.

Preparation of water extract fractions of the gold dust mushroom fruiting body was prepared by adding water to the gold dust mushroom fruiting body and extracted for 4-8 hours at 60-100 ℃, preferably at 80 ℃ 6 hours. In the preparation of the organic solvent precipitate fraction of the fruiting body of Geumsaeng Sichuan mushroom, 30-70% of cold acetone was added to the water extract, followed by precipitating and lyophilization by centrifugation. Preferably, 50% of acetone was added to prepare an organic solvent precipitate.

SPF ICR mice used as experimental animals were sold by Korean Animals Co., Ltd. in Kyung Hee University's environmental safety cabinet (Myungjin model), temperature 22 ± 0.5 ℃, humidity 55 ± 5%, exhaust 10 ~ 20 times / h , Fluorescent lamp 12h cycle, adjusted to the environment of light intensity of 300 ~ 500 lux and used for 1 week after acclimatization.

In this step, the acute oral toxicity test of the extract of S. aureus was carried out in accordance with the toxicity test standard of drugs such as the 1996-8 (April 16, 1996) notification of the Food and Drug Safety Headquarters. The experimental animals used for the acute oral toxicity test were SD rats weighing 80 to 90 g at 4 weeks of age, and 70 rats were purchased for 3 days in the animal room. General symptoms were visually observed during the acclimation period, and 60 healthy female cancers (30 females and 30 females) were used for the test. Separation of animal groups was based on the weight of the animals determined to be healthy during the accretion period, and evenly distributed animals of similar weight in a random manner by five animals per male and female group. Individual identification of animals was carried out by the hair coloring marking method.

The SD rats were raised in an animal room set at a temperature of 23 ± 3 ° C., a relative humidity of 50 ± 10%, an illumination time of 12 hours, a ventilation frequency of 10-20 / hr, and an illuminance of 150-300 Lux. During the test period, the temperature and humidity of the animal room were kept constant by automatic temperature and humidity control, and there was no change that would affect the test as a result of environmental monitoring. The breeding box used during the purifying and testing periods was plastic breeding box, which was 220W × 410L × 200Hmm and accommodated in 5 SD rats.

The feed used for the acute oral toxicity test was a solid feed for experimental animals, and the daily supply was limited to 200g. Feed contaminant identification was based on data provided by the supplier, and no factors affecting the test were found. Water was given a negative drinking water and the daily salary was limited to 480ml.

In the acute oral toxicity test, the dose of Geumsaek mushroom extract was very weak in the preliminary experiments. Therefore, the limiting dose was applied to the highest dose of 2,000mg / kg. 1,000 mg / kg and 500 mg / kg were administered respectively.

The significance test for the acute oral toxicity test was carried out by Dunnett's test, a multivariate comparison method by one way analysis of variance.

Hereinafter, the specific method of the present invention will be described in detail with reference to Examples, but the scope of the present invention is not limited only to these Examples.

Example 1 Preparation of Extracts of Artificially Cultivated Geumsa Sacral Mushroom Fruits

Extraction of artificial cultivated gold sand mushrooms was added to about 500g of water 5 times, extracted at 80 ℃ for 6 hours, filtered and the residue was repeatedly extracted. The extracts were combined, dialyzed and lyophilized to obtain a water extract fraction. In addition, the precipitate obtained by adding 50% cold acetone to the water extraction fraction was centrifuged and lyophilized to obtain an organic solvent precipitate fraction.

Example 2: In vitro Cytotoxicity Study of Artificially Cultivated Geum-Sang Mushrooms Fruiting Body

The present invention investigated the cytotoxicity against normal cells (MA-104) and cancer cells in vitro by using artificially cultured Phellinus linteus water extract or organic solvent precipitation fraction. Cancer cells used SNU-C4, A-549, P-388, L-1210, 10% FBS, 1% antibiotics-antimycotics and 2.2 g / L sodium bicarbonate ) Was cultured in supplemented RPMI 1640 medium. SNU-C4 and A-549 were treated with 0.25% trypsin to collect the cells from the flask and the cells were seeded at 3 × 10 ⁴ cells / well and 180 μl were dispensed into 96 wells. The cell line was stabilized in a CO ₂ incubator saturated with 5% CO ₂ at 37 ° C. for 24 hours. P-388 and L-1210 were stabilized in a CO ₂ incubator saturated with 5% CO ₂ for 2 hours by dispensing 180 μl by adjusting the cell number to 4 × 10 ⁴ / well. 20 μl of autoclaved samples were added to the cells at a concentration of 10 mg / ml, and the autoclave sterilized mushroom extract was added to the final concentration of the sample to 1 mg / ml, followed by 5% CO ₂ at 37 ° C. Incubated for 48 hours in a saturated CO ₂ incubator. After incubation, add 20 mg / ml of MTT reagent at 50 ㎖ per well and react in a CO ₂ incubator for 4 hours, remove the supernatant, add 100 μl of DMSO to the remaining precipitate, and absorbance with ELISA reader at 580 nm. Was measured. Adriamycin was used as a control drug.

As a result, as shown in Table 1, both water extract fractions and organic solvent precipitation fractions of artificially grown Geumsaeng mushrooms compared to normal cells (MA-104) showed very low toxicity or little effect on cancer cells.

Cytotoxicity against Cancer Cell Lines sample IC ₅₀ A549 SNU C4 L1210 P388 MA-104 WE 0.5 〉 0.5 0.38 〉 0.5 〉 0.5 OSP 0.5 〉 0.5 0.5 〉 0.5 〉 0.5 Adriamycine 0.05 0.02 0.01 0.001 0.5 WE: Water extract fraction of gold sand mushroom, OSP: Acetone sediment fraction of gold sand mushroom A549: Human lung cancer cell, SNU C4: Human colon cancer cell, L1210: Lymphocytic leuemia cell of mouse P388: Mouse lymphoid Neoplasm cells, MA-104: Macacus rhesus monkey kidney cells

Example 3: In vivo anticancer activity of Geumsaeng mushroom extract

In the present invention, the anticancer activity of the gold-grown mushroom (Phellinus linteus) artificially incubated in vivo was investigated. That is, the anticancer activity was investigated by inducing the ascites cancer and solid cancer in mice using sarcoma 180 and then administering the extract of Geumsaeng situation. First, the anticancer activity of gold dust mushrooms against ascites cancer was determined by taking ascites fluid from mice transplanted with sarcoma 180 (2 × 10 ⁷ cells / ml), suspended in physiological saline, and centrifuged at 2000 rpm. It was then washed twice and stained with 0.4% trypan blue to prepare 2 × 10 ⁷ cells / ml. All of the above operations were performed at 4 degreeC. The prepared sarcoma 180 was intraperitoneally administered 0.1 ml to 50 ICR mice (male 25g) and one group was divided into five groups of 10 rats. As a control of sarcoma 180, physiological saline was administered intraperitoneally to group 1, 100mg / kg of gold mushroom extract in group 2, and 10mg / kg of gold mushroom extract to group 3 in group 2. 100 mg / kg organic solvent precipitation fraction of Kumquat mushroom extract is included in Group 5 and 10mg / kg organic solvent precipitation fraction of Kumquat mushroom extract is contained in abdominal cavity for 7 days from the day of transplantation of sarcoma 180. Administered. Ten mice in the normal control group were compared with the survival rate while being bred for 40 days with only saline without administration of sarcoma 180. To measure anticancer activity against solid cancer, sarcoma 180 was prepared in the same manner as that for ascites, and 50 ml of ICR mice were injected into the orbit by implantation into the orbit and 10 groups of 3 animals were injected. Divided into groups. Group 1 contains physiological saline, group 2 contains gelatinous water extract, 25mg per unit weight of mouse, and group 3 contains organic solvent precipitation fraction of geumsae mushroom extract, 25mg per unit weight of mouse. Was administered intraperitoneally for 7 days from the day of implantation. Ten mice in the normal control group were administered only saline without administration of sarcoma 180, and were weighed by taking solid cancer cells in the orbit after slaughtering for 20 days.

As a result, all cases of ascites were killed within 30 days in the control group and survival time was 16.4 days. However, as shown in Table 2, when administered the water extract fractions of artificially cultured Geumsaek situation, there was more than 30% life extension effect at 10mg / kg per unit weight compared to the control group, survival rate was 20%. In the case of 100mg / kg body weight, 50% or more life extension effect and 20% survival rate were shown. When 10mg / kg of the organic solvent precipitate fraction of the water extract of Geumsaeng mushroom was extracted, it showed more than 40% life extension effect and 40% survival rate. When 100mg per unit weight was administered, there was no change in the survival rate. It showed a tendency to decrease. These results show the potential for toxicity at higher doses.

Anticancer Activity of Geumsaeng-mushroom Extracts in ICR Mice Induced Ascites Cancer with sarcoma 180 group Dose (mg / kg body weight) Survival rate (%) Survival Day Control 0 100 - Group 1 0 0 16 ± 4.7 ^a Group2 100 20 25 ± 4.7 ^b Group3 10 20 22 ± 3.0 Group 4 100 40 20 ± 4.0 Group 5 10 40 23 ± 1.2 ^b Group 1: Treated with sarcoma 180, Group 2, 3: Administering water extract fractions of gold dust mushroom Group 4, 5: Administration of acetone precipitate fraction of gold dust mushroom ^a : Mean ± Standard deviation ^b : Statistical comparison with control Significance (P <0.05)

In the case of inducing solid cancer in the mice, the weight of the cancer cells was 3.6g after 21 days of cancer cell administration in the control group. However, when the organic solvent precipitate fraction was administered, it showed a significant inhibitory effect of 70% or more. Geumsaeng mushroom had no direct anticancer effect against cancer cells, but showed anticancer effects when cancer cells were transplanted in vivo.

Anticancer Activity of Geumsa Sacral Mushroom Extract from ICR Mice That Caused Solid Cancer with sarcoma 180 group Dose (mg / kg body weight) Cancer Cell Weight (g) Control 0 0 Group 1 0 3.6 ± 1.80 ^a Group 2 25 3.5 ± 2.10 Group 3 25 1.1 ± 1.20 ^b Note 1: Group 1: treated with sarcoma 180, group 2: administration of water extract fractions of S. aureus group 3: administration of acetone precipitate fractions of S. aureus ^a : mean ± standard deviation ^b : statistical significance compared to control p <0.05)

Example 4: Acute Toxicity Study of Oral Administration of Geumsa Sacral Mushroom Extract

After extracting the extract of Geumsa Shang mushroom, it was dissolved in tertiary distilled water, and acute toxicity was investigated by administering 500 mg / kg to 2,000 mg / kg of body weight per SD rat. Rats were fasted one night before administration and administered directly into the stomach using an oral needle. The oral administration was selected as one of the predictive routes of clinical application, and the dosage was calculated based on the weight of the day of administration. After the oral administration of Geumsa Sacral Mushroom extract to SD rats, the general symptoms of animals, the presence or absence of dead animals, weight change, feed intake and drinking volume were observed.

General symptoms and death of animals were observed every hour from 1 hour to 12 hours after administration on the day of administration, and changes in general symptoms, poisoning symptoms and the presence of dead animals were observed at least once daily from the day after administration to 14 days. The weight change and feed intake change of the test animals were measured every day before and after the start of the administration for all subjects used in the test. The average intake was investigated by dividing the change in g number from the previous day after the population. The change of drinking water was measured by dividing the change by the number of individuals by feeding 480ml in the same manner as the feed intake. Further experiments were conducted by administering 4,000 mg / kg to rats with a limit of 5,000 mg / kg as reference to the Japanese Ministry of Health, Labor and Welfare's Commentary.

As a result of the experiment, the mortality rate of rats orally administered with S. aureus mushroom extract did not die during the experimental period.

Mortality of SD Rats after Oral Administration of Geumsa Sacral Mushroom Extract gender Dosage 0 days 1 day 2 days 3 days ...... 13th 14 days cock 0 0/5 ^* 0/5 0/5 0/5 0/5 0/5 0/5 500 0/5 0/5 0/5 0/5 0/5 0/5 0/5 1,000 0/5 0/5 0/5 0/5 0/5 0/5 0/5 2,000 0/5 0/5 0/5 0/5 0/5 0/5 0/5 female 0 0/5 0/5 0/5 0/5 0/5 0/5 0/5 500 0/5 0/5 0/5 0/5 0/5 0/5 0/5 1,000 0/5 0/5 0/5 0/5 0/5 0/5 0/5 2,000 0/5 0/5 0/5 0/5 0/5 0/5 0/5 [Note] ^* : Number of dead animals / number of experimental animals per unit group Dose unit: mg / kg

As a result of observing the clinical symptom for 14 days after the administration of Geumsaeng mushroom extract in rats, as shown in Table 5, there was no change in both cancer and male rats.

Changes in Clinical Symptoms of SD Rats after Oral Administration of Geum-Sang Mushroom Extract gender Dosage Observation Observation period cock 0 normal 0-14 days after dosing 500 normal 0 to 14 days 1,000 normal 0 to 14 days 2,000 normal 0 to 14 days female 0 normal 0 to 14 days 500 normal 0 to 14 days 1,000 normal 0 to 14 days 2,000 normal 0 to 14 days Dosage unit: mg / kg

The higher the concentration of rats during the test period, the higher the increase in the first 3 days after administration, and the weight gain rate was lower than that of the control group during the entire test period. This phenomenon was more pronounced at higher concentrations in rats. In conclusion, as shown in Table 6, the experimental group to which oral administration of the extract of S. aureus showed lower body weight gain rate than the control group during the 14-day test period.

Changes in Body Weight of SD Rats after Oral Administration of Geumsa Sacral Mushroom Extract Dosage Control (M) Control (F) 2,000 (M) 2,000 (F) 1,000 (M) 1,000 (F) 500 (M) 500 (F) 0 days 86.0 82.0 82.2 81.6 88.8 84.0 87.2 82.4 1 day 98.0 94.0 92.6 91.2 100.4 94.2 99.6 98.6 2 days 109.5 106.0 104.0 101.4 108.0 106.2 107.6 105.8 3 days 123.5 120.0 116.0 107.2 119.8 119.2 116.8 115.8 4 days 133.5 126.0 118.8 114.8 128.2 127.8 123.6 121.6 5 days 144.0 132.0 121.6 118.8 131.6 130.6 129.4 128.2 6 days 149.0 136.0 129.6 124.6 136.0 135.0 134.8 132.8 7 days 154.5 140.0 134.4 129.6 142.4 142.6 140.4 139.6 8th 159.5 145.5 139.6 134.4 149.2 146.6 148.0 145.8 9th 163.5 151.0 146.4 138.6 152.2 150.0 153.0 151.2 10 days 167.0 158.0 150.2 142.2 158.4 156.2 159.2 158.2 11th 170.0 165.0 154.8 149.6 162.8 160.4 165.4 164.4 12 days 187.0 168.0 159.8 154.6 169.6 166.8 170.2 170.4 13th 183.0 147.0 162.4 160.0 174.2 171.6 178.6 179.2 14 days 191.5 181.0 166.8 164.4 180.4 178.8 184.8 158.6

As shown in Table 7, the feed intake rate was 0.8 times compared to the control group in the first 3 days after the administration. .

Changes in Feed Intake of SD Rats after Oral Administration of Geumsa Sacral Mushroom Extract Dosage 1 day 2 days 3 days 4 days 5 days 6 days 7 days 8th 9th 10 days 11th 12 days 13th 14 days Control (M) 13.0 14.0 16.0 18.5 20.3 24.5 28.1 31.0 32.3 33.0 33.0 34.7 35.0 36.0 Control group (F) 12.0 13.0 13.0 14.7 16.3 17.0 18.6 19.0 19.0 20.2 20.0 22.0 23.4 24.0 2,000 (M) 9.0 10.0 10.5 13.0 13.6 15.0 15.0 14.7 17.0 17.2 18.4 19.6 20.0 22.0 2,000 (F) 8.9 9.2 10.0 10.0 11.2 11.0 11.0 13.3 13.0 15.2 16.0 17.5 19.0 20.0 1,000 (M) 12.0 13.0 13.4 12.3 14.0 14.8 15.0 16.0 18.0 17.8 18.0 19.0 19.7 21.3 1,000 (F) 11.2 12.0 12.0 13.8 14.0 14.5 15.0 15.0 14.0 15.4 16.7 18.0 19.0 20.0 500 (M) 13.0 12.0 13.4 14.0 15.4 16.1 15.9 16.4 17.4 17.0 18.3 18.9 20.0 21.3 500 (F) 9.8 10.2 11.0 12.4 13.8 15.2 16.0 16.0 15.8 17.0 18.2 19.0 19.8 22.0 (Note) M: Male rat F: Female rat Dose unit: mg / kg, Feed intake unit: g

The change of drinking water after acute toxicity test is shown in Table 8. Compared with the control group, the average drinking volume of the first three days was about 2 times higher than 2,000 mg / kg of the gold mushroom extract, about 1.8 times higher than 1,000 mg / kg, and 1.5 times higher than 500 mg / kg. It was found to be about twice as many, and from day 4 it was similar to the control group.

Changes in Drinking Water Levels of SD Rats after Oral Administration of Geumsa Sacral Mushroom Extract Dosage 1 day 2 days 3 days 4 days 5 days 6 days 7 days 8th 9th 10 days 11th 12 days 13th 14 days Control (M) 8.3 10.0 11.3 16.5 18.4 20.6 24.6 27.8 29.2 31.5 34.9 38.5 42.5 47.9 Control group (F) 8.0 10.2 12.0 15.4 19.2 21.3 24.5 27.9 29.8 30.8 33.8 37.9 42.0 48.0 2,000 (M) 22.0 24.3 23.5 14.0 15.9 18.6 21.5 24.0 26.4 29.0 32.1 36.9 40.8 47.0 2,000 (F) 19.6 20.4 25.0 18.2 18.3 20.0 21.3 24.0 24.9 28.0 30.7 34.8 39.5 45.3 1,000 (M) 20.1 21.4 22.0 17.9 19.5 21.1 22.4 24.6 24.8 25.9 27.8 28.2 31.5 37.5 1,000 (F) 19.4 18.4 19.5 17.2 18.4 20.5 21.8 23.5 25.3 25.8 28.0 30.0 31.2 34.8 500 (M) 15.4 15.0 15.6 14.8 16.8 18.9 20.5 21.8 23.5 24.7 27.0 29.5 31.0 34.0 500 (F) 15.0 14.0 15.4 15.0 16.4 17.8 19.4 20.8 21.4 23.0 24.7 25.8 27.0 29.8 (Note) M: Male rat F: Female rat dose unit: mg / kg, drinking water unit: ml

All male and female rats used as experimental animals were necropsied 14 days after administration. As shown in Table 9, no abnormality was observed by visual observation.

Autopsy Results of SD Rats 14 Days After Oral Administration of Geumsa Sacral Mushroom Extract gender Dosage Autopsy findings frequency Male cut 0 normal 0/0 ^* 5/5 500 normal 0/0 5/5 1,000 normal 0/0 5/5 2,000 normal 0/0 5/5 female 0 normal 0/0 5/5 500 normal 0/0 5/5 1,000 normal 0/0 5/5 2,000 normal 0/0 5/5 [Note] ^* : Number of rats showing abnormal findings / Number of rats per unit group Dose unit: mg / kg

In the case of additional experiments with high concentrations of the extracts of S. aureus, all the animals survived for 14 days and the necropsy showed no abnormalities.

Taken together, the Geumsa Sacral Mushroom extract was tested in acute oral toxicity test with 5 rats and 5 rats at doses of 0, 500, 1,000, 2,000, 4,000 and 5,000 mg / kg for SD rats. As a result of the experiment, the high-dose group, that is, 2,000mg / kg or more, was considered to cause dry mouth considering the negative drinking amount for 3 days after the administration. No unusual changes in feed intake and autopsy findings were observed for LD ₅₀ so that the LD ₅₀ value was thought to be more than 5,000 mg / kg for both male and female rats. Therefore, it was confirmed that Geumsa Sacral Mushroom extract is clinically safe to use.

As described above, the water extract fraction or the organic solvent precipitate fraction of the Geumsaeng situation mushroom fruiting body of the present invention did not show cytotoxicity against cancer cell lines in vitro, but it enhanced the immune ability of the living body in vivo and ascites cancer. It has the effect of increasing the survival rate and prolonging the survival rate of the mice that caused the cancer and solid cancer, inhibiting the growth of solid cancer, and providing the composition for the prevention and treatment of cancer, which contains extracts of the fruiting body of the gold barberry. This is a very useful invention in the biomedical industry because it is clinically safe to use.

Claims (2)

Cancer preventive and therapeutic composition comprising the extract of fruiting body of the gold dust situation as an active ingredient. (a) adding water to the fruiting body of artificially grown gold sand mushrooms, extracting and filtering for 4-8 hours at 60-100 ° C., extracting the residue repeatedly, and dialysis and lyophilizing to prepare a water extract fraction; (b) centrifugal separation of the precipitate obtained by adding cold acetone to 30 to 70% to the water extraction fraction of step (a) and lyophilization to prepare an organic solvent precipitate fraction; Extraction method of fruiting body.