KR20010086948A - Pharmaceutical composition for the prevention and treatment of hepatoma - Google Patents

Abstract

PURPOSE: Provided is a novel pharmaceutical composition which shows excellent treatment and prevention effects on liver cancer without toxicity and has pharmaceutical stability. CONSTITUTION: The pharmaceutical composition comprises larvae of Holotrichia diomphalia Bates(scarabaeidae); and fruits of Gardenia jasminoides Ellis, Scutellaria baicalensis Georgi(Labiatae) and dried roots of Bupleurum falcatum Linne(Umbelliferae), in particular, in a weight ratio of 0.7-1.5 : 1-2 : 1 : 1-2. The composition can be used in an amount of 15 to 300mg and administered in formulation types of tablet, powder, granule, solution, syrup or capsule. The composition is prepared by following steps of: mixing the active ingredients, and then extracting them with organic solvents while refluxing; filtering the extract to obtain filtrate; further extracting and filtering twice the residue to obtain further filtrate; combining the filtrates; concentrating under pressure to prepare the extracts; and adding a pharmaceutical acceptable carrier to the extracts to form the formulation in a desired type.

Description

Pharmaceutical composition for the prevention and treatment of liver cancer {PHARMACEUTICAL COMPOSITION FOR THE PREVENTION AND TREATMENT OF HEPATOMA}

The present invention is prepared having a prophylactic and therapeutic effect against liver cancer ( ), And a pharmaceutical composition for the prevention and treatment of new liver cancer containing gardenia, gold and shiho as an active ingredient.

An anticancer agent binds and acts directly on DNA in a cell to block DNA replication, transcription, and translation, or interfere with nucleic acid synthesis and inhibit cell division by interfering with metabolic pathways of nucleic acid synthesis. Drugs exhibiting cytotoxicity against are generically referred to. Anticancer drugs are classified into six groups based on their mechanism of action and chemical structure: alkylating agents, anticancer antibiotics, metabolic antagonists, hormones, cell division inhibitors, and other groups.

1) Alkylating agents-busulfan, melphalan, chlorambucil, cyclophosphamide and the like are known,

2) Anti-cancer antibiotics-doxorubicin, daunorubicin, bleomycin, mithramycin, dactinomycin, mitomycin-C, and mitomycin C. Belong to,

3) metabolic antagonists-cytosine arabinoside, 6-mercaptopurine, 5-fluorouracil, 5-fluorouracil, methotrexate, etc. are known,

4) hormones-androgens (androgens), progesterone, estrogen (estrogens), corticosteroids, tamoxifen (tamoxifen), etc.

Others-vinblastine, vincristine, teniposide, etoposide, BCNU, CCNU, cisplatin, taxol, etc. Known.

However, anticancer drugs generally have very narrow therapeutic and toxic doses, so if the dose is slightly increased, the destruction of cancer cells is much more severe, but serious side effects occur.If a small amount is used, the anticancer effect is lost, resulting in minimal side effects. The principle is to administer the maximum allowed dose.

Nevertheless, there are differences in the degree of chemotherapy, but the side effects (toxicity) occur in almost all patients, and as a result, the patient's pain may be severe or addicted to death. The development of anti-cancer drugs with a few) is an urgent task in biotechnology and medicine.

Accordingly, the present invention is to provide a pharmaceutical composition for the prevention and treatment of new liver cancer with low side effects (toxicity).

Pahyeol the conventional shot, hanger, and sangyeol tongyu the animal herbal manufacture in action [gold slugs (Holotrichia diomphalia Bates) and larvae of insects closely related (gumbengyigwa scarabaeidae)]; Gardenia in cheongyeol reconciliation, yanghyeol detoxification is [(Gardenia Gardenia jasminoides Ellis or other fruit of dongsok plants (Rubiaceae Rubiaceae)]; antipyretic, diuretic, antibacterial, antiviral, antifungal, relax blood pressure, blood sugar increase, yidam action Botanical herbal medicines such as golden backed roots (skinned roots of Scutellaria baicalensis Georgi (Lacaceae Labiatae )] have been used as a single ingredient in traditional herbal medicines for liver disease.

In addition, the herbal medicine Shiho (dried roots of Bupleurum falcatum Linne) has been widely used for pleurisy and chest pain due to antipyretic and analgesic effects.

However, although the herbal medicines such as gardenia, gold, and sea tiger are among the herbal medicines with safety, the manufacture of slug larvae has been reported to be highly toxic. In experiments on the effects of hepatic impairment [Journal of Pharmacognosy, 22 (1), 36-44 (1991)], the aqueous ethanol extract of the preparation had an effect on carbon tetrachloride-induced hepatic impairment. It has been reported that a toxicity of up to 10 mortality was recognized at 1,000 mg / kg / kg and intraperitoneal administration.

Therefore, in the present invention, in order to remove such toxic substances first, the numerical value was first determined based on the numerical method currently available in the traditional Chinese medicine, and then the prescription of the remaining three herbal medicines was based on the herbal medicine.

In other words, these four herbal medicines (manufacture, gardenia, gold and Siho) are selected, washed, cut, sterilized and dried. The extract of the combination was prepared according to the various manufacturing methods of X, and the drug was tested for efficacy and toxicity of liver cancer cells with a certain amount of X. As a result, it was confirmed that there was no toxicity and had an excellent effect on the treatment and prevention of liver cancer. This invention was completed.

Accordingly, the present invention aims to provide a novel pharmaceutical composition having less toxic or side effects than the manufactured (gumbang) single agent and having a superior therapeutic and prophylactic effect of liver cancer.

The pharmaceutical composition for the prevention and treatment of liver cancer of the present invention contains an animal herbal medicine and a vegetable herbal medicine, gardenia, gold and shiho, and specifically, the extract of gardenia, gardenia, gold and shiho is 0.7-1.5: 1 to 1-2. A pharmaceutical composition for preventing and treating liver cancer, which is contained in a weight ratio of 1: 1 to 2 :.

In the manufacturing method of the pharmaceutical composition of the present invention, after the numerical processing of the preparation, gardenia, gold and shiho, respectively, they are mixed by a predetermined amount, extracted with an organic solvent, and then obtained under reduced pressure and concentration to obtain a pharmaceutically acceptable carrier for each formulation. It is formulated by adding.

First of all, the production of strong toxic effects is carried out by the method already established by the emulsification method, that is, 1) removing mud from the sieve and washing it to dry in the daylight (brain poszaron), or 2) glutinous rice after drying. Roast until it turns black, and remove hair growth around the mouth and body, cut into 3-4 pieces (agreement, medical introduction), or 3) put it in hot water, wash it with purified water, and dry it in daylight. One of the animal animal medicines was selected and quantitatively prepared into powder or cut by adjustment.

Gardenia, Golden and Siho are each selected by washing, drying, cutting and sterilization of commercially available herbal medicines, and then powdered or adjusted to prepare each herbal medicine as a pharmaceutical.

These preparations, gardenia, golden and wild medicinal herbs were mixed at a weight ratio of 0.7 to 1.5: 1 to 2: 1: 1 to 2, and then extracted using various extracting organic solvents. The extraction solvent was extracted three times using organic solvents such as 70 saturated ethanol, ethanol, methanol, and ethyl acetate, and the extracts were combined to prepare X using lyophilization or reduced pressure rotary concentrator. The extracted extract was found to have a good yield, the best drug against liver cancer, and no toxicity at all. This fraction was used as an active ingredient for the prevention and treatment of liver cancer. In the present invention, when the mixing ratio is 0.7 or less, the effect on liver cancer treatment is questioned, and when it is 1.5 or more, toxic expression is concerned. Gardenia, Golden and Shiho Herbal Medicines were also preferred in terms of drug efficacy and preparation in the above ratio within the range of normal dose.

Furthermore, the pharmacological effect determination experiment of the pharmaceutical composition according to the present invention was carried out. In the anticancer effect test by the extract of extract containing the compound prepared in macrophages isolated from the mice, the extract of the complex containing extracts prepared in macrophages using MTT detection method Treatment of S-1 and S-2 at 0.1, 1, 10 and 100 μg / ml respectively and incubation with cancer cells for 18 hours showed that the macrophage cancer cell toxicity of the negative control group was 37.7 but 0.1 μg of the experimental group S-1. Cancer cell toxicity of 48.74, 55.74, and 70.57 at / ml, 1 μg / ml and 10 μg / ml was shown to be concentration dependent. At 10 ㎍ / ml, the cytotoxicity of macrophages was 87.18 more than that of the negative control, and higher than that of the positive control (61.72). Experimental group S-2 increased the cytotoxicity of macrophages by 36.45 only at 10 ㎍ / ml compared to the negative control group, which showed lower activity of macrophages than experimental group S-1. The preparation single showed 45.92 only at 10 μg / ml, which was lower than the cytotoxicity of the extract containing the combination at the same concentration. Positive control group was treated with lipopolysaccharide (1 μg / ml) and gamma-interferon (10 unit / ml) that induce anticancer activity.

From the results of these experiments, the experimental groups showed a superior effect than the monoliths of manufacture, it can be said that the treatment effect for liver cancer is excellent.

The pharmaceutical composition of the present invention can be used alone, 70 hydrous ethanol extract extract, methanol extract extract and ethyl acetate extract extract in the extract extract, the formulations are tablets, powders, granules, It can be prepared by using a liquid, syrup, capsule, or the like.

In the pharmaceutical composition of the present invention, it is preferable that the general adult divides the amount of 15 to 300 mg once or several times a day.

Hereinafter, the present invention will be described in detail through Examples and Experimental Examples, and the present invention is not limited to these Experimental Examples and Examples.

(Reference Example 1)

Preparation of 70 hydrous ethanol extract extract (S) from the production (slugs)

100 g of 70 g of ethanol was added to the mixture, and the mixture was heated and extracted for 6-8 hours in a reflux extractor, filtered, and 500 ml of 70 hydrated ethanol was added to the residue. The extraction was repeated three times, and the extract was extracted and combined with a pre-extracted extract.

(Example 1)

Preparation of composite extract extract (S-1)

190 g of commercially produced (Golden slug), 270 g of Gardenia jasmine, 270 g of Gold, 270 g of Siho were mixed and adjusted, and 3 liters of 70 hydrated ethanol were added and heated and extracted for 6-8 hours in a reflux extractor. The extract is then cooled and filtered. In the same way, the extract obtained by two additional extractions was combined with the extract and then concentrated under reduced pressure at 40 ° C. in a vacuum condenser to obtain 350 g (yield of 0.7: 1: 1: 1 ratio of manufacturing, gardenia, gold and shiho) (yield 35). Got.

(Example 2)

Preparation of composite containing extract extract (S-2)

250 g of commercially produced (Golden slug), 250 g of Gardenia, 250 g of Gold, 168 g of Gold, and 332 g of Shiho were each adjusted and mixed, followed by 3 liters of 70 hydrous ethanol, which were heated and extracted for 6-8 hours in a reflux extractor. The extract is then cooled and filtered. In the same way, the extract obtained by two additional extractions was combined with the extract, and then concentrated under reduced pressure at 40 ° C. in a vacuum condenser to obtain 340 g (yield: 1.5: 1.5: 1: 2 ratio of manufacturing, gardenia, gold, and shiho) (yield 34). Got.

(Example 3)

Preparation of composite containing extract extract (S-3)

Commercially purchased (Golden slug) 190 g, Gardenia 270 g, Golden 270 g, Shiho 270 g were mixed and adjusted, and 3 liters of methanol was added thereto, and the mixture was heated and extracted for 6-8 hours in a reflux extractor. After cooling, it is filtered. In the same way, the extract obtained by two additional extractions was combined with the extract, and then concentrated under reduced pressure at 40 ° C. in a vacuum condenser to obtain 360 g (yield of 0.7: 1: 1: 1 in manufacturing, gardenia, gold and shiho). )

(Example 4)

Preparation of composite extract extract (S-4)

250 g of commercially produced (gold slugs), 250 g of gardenia, 168 g of gold, 332 g of shiho were each adjusted and mixed, and then 3 liters of ethanol was added and heated and extracted in a reflux extractor for 6 to 8 hours. After cooling, it is filtered. In the same way, the extract obtained by two additional extractions was combined with the extract, and then concentrated under reduced pressure at 40 ° C. in a vacuum condenser to obtain 350 g (yield of 1.5: 1.5: 1: 2 of manufacturing, gardenia, gold and shiho) (yield 35 )

(Example 5)

Preparation of composite extract extract (S-5)

190 g of commercially produced (gold slugs), 270 g of gardenia, 270 g of gold, 270 g of shiho were each adjusted and mixed, 3 liters of ethyl acetate was added, heated and extracted with a reflux extractor for 6-8 hours. The extract is cooled and then filtered. In the same way, the extract obtained by two additional extractions was combined with the extract, and then concentrated under reduced pressure at 40 ° C. in a vacuum condenser to obtain 170 g (yield of 0.7: 1: 1: 1 in manufacturing, gardenia, gold and shiho) (yield 17 )

(Example 6)

Preparation of composite extract extract (S-6)

250 g of commercially produced (gold slugs), 250 g of gardenia, 168 g of gold, 332 g of shiho were each adjusted and mixed, 3 liters of ethyl acetate was added, heated and extracted with a reflux extractor for 6-8 hours. The extract is cooled and then filtered. In the same way, the extract obtained by two additional extractions was combined with the extract, and then concentrated under reduced pressure at 40 ° C. in a vacuum condenser to obtain 160 g (yield of 1.5: 1.5: 1: 2 of manufacturing, gardenia, gold and shiho). )

(Composition Example 1)

Solution

-In 500 ml

<Example 1> or 150 mg of extracts containing the preparation of <Example 2>

Isomerized sugar 55 g

L-sodium glutamate 3.4 g

Phosphoric Acid 1.5 g

Herb Essence

0.5 g sodium benzoate

0.5 g of sodium dehydroacetate

Tablet quantity

After dissolving isomerized sugar, sodium benzoate and sodium dehydroacetate in a predetermined amount of purified water, and adding the extract extract of the composite containing Preparation of Example 1 or 2 to make a homogeneous solution, sodium glutamate and phosphoric acid, flavoring, flavoring Add to mix. The final dose was applied to 500 ml, and then filtered and the filtrate was filled in 50 ml bottles in a conventional manner and sterilized to prepare a solution containing 15 mg of the combined extract extract containing the preparation per bottle.

(Composition Example 2)

Granules

-In 300 g

<Example 1> or 1.5 g of the extract containing the preparation of <Example 2>

240 g lactose

45 g of caramel powder

1.5 g of hydroxypropyl cellulose

Purified water

1) First, 240 g of lactose and 45 g of caramel powder are stirred and mixed evenly.

2) 1.5 g of pre-hydroxypropyl cellulose (HPC) was added to 30 ml of ethanol and completely dissolved. Then, 1.5 g of the extract of the composite containing the preparation of <Example 1> or <Example 2> was added to this solution, and gradually 1 Stir homogeneously for hours.

3) To the mixed powder prepared in advance in the above 1) was slowly added while stirring the main chemical solution of 2), mixed until homogenized, and then added by adding a suitable amount of purified water, homogenized again, and then the preparation of the pharmacopoeia granules According to the method, granules were prepared such that 15 mg of the extract-containing complex extract was contained in 1 sachet (3 g).

(Composition Example 3)

refine

A tablet containing 15 mg of the extract containing the composite preparation was prepared by compression molding the granules prepared in Composition Example 2 per 1000 mg.

Experimental Example 1

Stability test according to the change over time of the composition (granule) containing the extract extract

1. Sample

The granular composition containing the preparation-containing complex extract extract prepared according to Example 1 was tested as a specimen.

2. Experimental method

1) Appearance

It was visually confirmed.

2) Confirmation test

When tested according to the quantitative method, the sample solution was confirmed to show a peak at the same retention time as the standard solution.

3) According to the herbal medicine test method of the general test method of the pharmacopeias.

4) Content test

The content of Baicalin in Composition Example 2 (granule) was analyzed by high performance liquid chromatography.

5) Preservation of Specimen

Sample preservation was tested at 40 ° C. (relative humidity 75) at room temperature and in LifeTesta.

3. Experimental Results

Each experimental data was the average of the results of three or more experiments (Table 1).

Table 1

Stability test (acceleration test) according to change of composition example 2 (granule) with time

Manufacturing number Storage condition item period Room temperature Manufacturing number Storage condition item period 40 ° C., 75 R.H. Constellation Confirm Ash () baicalin content Constellation Confirm Ash () baicalin content Pilot1 Start fitness positivity 3.29 99.87 Pilot1 Start fitness positivity 3.28 99.84 2 months 3.30 99.86 2 months 3.24 99.87 4 months 3.28 99.84 4 months 3.25 99.68 6 months 3.26 98.82 6 months 3.26 98.25 Pilot2 Start fitness positivity 3.25 99.80 Pilot2 Start fitness positivity 3.22 99.84 2 months 3.28 99.82 2 months 3.21 99.23 4 months 3.23 99.84 4 months 3.26 99.21 6 months 3.21 98.73 6 months 3.24 97.98 Pilot3 Start fitness positivity 3.26 99.92 Pilot3 Start fitness positivity 3.28 99.84 2 months 3.30 99.90 2 months 3.23 99.32 4 months 3.27 99.84 4 months 3.24 99.34 6 months 3.29 98.74 6 months 3.22 97.32

Experimental Example 2

Stability test according to the change over time of the composition (granule) containing the extract extract

1. Sample

The granular composition containing the extract containing the preparation containing the preparation prepared in Example 2 was tested as a sample.

2. Experimental method

1) Appearance

It was visually confirmed.

2) Confirmation test

When tested according to the quantitative method, the sample solution was confirmed to show a peak at the same retention time as the standard solution.

3) According to the herbal medicine test method of the general test method of the pharmacopeias.

4) Content test

Composition Example 2 The content of Baicalin in the granules was analyzed by high performance liquid chromatography.

5) Preservation of Specimen

Sample preservation was tested at 40 ° C. (relative humidity 75) at room temperature and in LifeTesta.

3. Experimental Results

Each experimental data was the average of the results of three or more experiments (Table 2).

Table 2

Stability test (acceleration test) according to change of composition example 2 (granule) with time

Manufacturing number Storage condition item period Room temperature Manufacturing number Storage condition item period 40 ° C., 75 R.H. Constellation Confirm Ash () baicalin content Constellation Confirm Ash () baicalin content Pilot1 Start fitness positivity 3.22 99.77 Pilot1 Start fitness positivity 3.26 99.93 2 months 3.29 99.82 2 months 3.23 99.89 4 months 3.23 99.81 4 months 3.24 99.34 6 months 3.25 98.78 6 months 3.27 98.21 Pilot2 Start fitness positivity 3.26 99.91 Pilot2 Start fitness positivity 3.23 99.91 2 months 3.23 99.90 2 months 3.24 99.88 4 months 3.26 99.82 4 months 3.25 99.76 6 months 3.24 98.33 6 months 3.22 97.49 Pilot3 Start fitness positivity 3.25 99.97 Pilot3 Start fitness positivity 3.27 99.92 2 months 3.23 99.92 2 months 3.23 99.89 4 months 3.21 99.92 4 months 3.25 99.28 6 months 3.23 98.53 6 months 3.26 97.28

Experimental Example 3

Anticancer Effect by Extract Extract from Complexes in Macrophages Isolated from Mice

1. Experimental Materials and Methods

1) Experimental animals and reagents

C57 / BL6 mice used in this experiment were purchased from Charles River Breeding Laboratories, Japan, and all reagents were purchased from Sigma (St Louis, MO) unless otherwise noted.

2) Isolation of hepatic macrophages

Macrophages were isolated using the following method. Syringe was put into portal vein and perfused at 10 ml / min rate of Hanks' balanced salt solution (HBSS) without Ca and Mg (200 IU / ml penicillin, 200 ㎍ / ml streptomycin) and perfused with 10 ml HBSS. After a third perfusion with 0.05 collagenase and HBSS, the liver was removed aseptically. The liver was cut and incubated for 45 minutes at 37 ℃, the tissue was passed through a 150 mesh screen with 7 ml of collagenase solution and incubated for 45 minutes at 37 ℃. Liver fluid was filtered using gauze and centrifuged twice at 500 g for 5 minutes. Pellet was suspended in HBSS containing 5 FBS and centrifuged twice at 500 g for 5 minutes. Parenchymal cells were destroyed by repeated pipetting with HBSS containing 0.05 trypsin, and centrifuged twice at 50 g in HBSS containing 5 FBS for 2 minutes. Nonparenchymal cells were collected by centrifugation at 500 g for 10 minutes, resuspended in 3 ml of RPMI containing 10 FBS, washed twice, and resuspended in 1 ml of RPMI to calculate the cell number. Cells were placed in a plate and allowed to attach for 2 hours, culture was removed, and adherent cells were collected by treatment with trypsin.

3) Treatment of macrophages with the manufacturing single and the extract extract containing the preparation of the present invention

Macrophages were inoculated in a 96 well tissue culture plate (Nunc, Denmark) at a concentration of 2 × 10 ⁵ cells / well and incubated for 2 hours, and then non-adhered cells were treated with 10 fetal bovine serum (FBS) and penicillin (100 IU / ml). Washed twice with RPMI-1640 (Gibco, Grand Island, NY) (RPMI-FBS) containing and streptomycin (100 μg / ml), extracts of various concentrations of manufacturing singles (S) and extracts containing the preparation (S-1). ) And (S-2), respectively, were added to carry out cytotoxicity test.

4) Cytotoxicity test of macrophages against cancer cells

The isolated macrophages were added with various concentrations (0.1, 1, 10, 100 ㎍ / ml) of the preparation single extract and the extract containing the complex, respectively, and in combination with cancer cells (B16-F10) in a 95 air / 5 CO ₂ incubator at 37 ° C. After incubation for 18 hours, 25 μl of MTT (2 mg / ml) was added and incubated for another 4 hours. Centrifuge the plate at 200 g for 5 min, remove supernatant leaving only 50 μl, add 150 μl DMSO to dissolve formazan crystals, shake for 10 minutes and measure OD at 540 nm using a microplate reader. The cytotoxicity was calculated using the formula.

OD (macrophages + cancer cells)-OD (macrophages)

Cytotoxicity () = 100---------------------------------------- × 100

OD (cancer cell)

2. Experimental results

Macrophages have been reported to be activated by a variety of substances and increase their resistance to cancer when administered to animals. In order to determine the effect of macrophages on anti-cancer action, macromolecules were prepared by using the MTT screening method for the preparation of single, mixed extracts (S-1) and (S-2) of 0.1, 1, 10 and 100 μg / Each cell was treated with ml and incubated with cancer cells for 18 hours. Lipopolysaccharide (1 μg / ml) and gamma-interferon (10 unit /) were used as positive controls to induce anticancer activity. ml). It is shown in Table 3 compared to the negative control group treated only with the culture solution. At high concentrations (100 μg / ml) of the mixtures used in the experiments, all showed cytotoxicity, making it difficult to measure anticancer activity. The macrophage cancer cytotoxicity of the non-sampled negative control group was 37.7, but the combined extract extract (S-1) containing 48.74, 55.74 and 70.57, respectively, at 0.1 µg / ml, 1 µg / ml and 10 µg / ml, respectively. Is shown to be concentration dependent. At 10 ㎍ / ml, the cytotoxicity of macrophages was 87.18 more than that of the negative control and higher than that of the positive control (61.72). Extract-containing complex extract (S-2) increased the cytotoxicity of macrophages by 36.45 only at 10 ㎍ / ml compared to the negative control group, which showed lower activity of macrophages than experimental group (S-1). On the contrary, the production single compound showed 45.92 only at 10 ㎍ / ml, which is lower than the cytotoxicity of the extract containing the complex at the same concentration, and thus the effect of inducing the anticancer activity of the macrophage-derived complex extract (S-1) Is estimated to be better.

Table 3

Cancer Cytotoxicity of Macrophages Induced by Extract Extracted from Ginseng

sample Cytotoxicity () Cytotoxicity Growth Rate () ^T Negative control group (culture amount) 37.70 Positive control group (LPS + IFN-Υ) 61.72 63.70 Compound Extract Extract (S-1) 0.1 μg / ml 48.74 ^* 29.27 1 μg / ml 55.74 ^* 47.86 10 μg / ml 70.57 ^* 87.18 Compound Extract Extract (S-2) 0.1 μg / ml 36.02 0 1 μg / ml 44.41 ^* 17.78 10 μg / ml 51.44 ^* 36.45 Manufacturing Single (S) 0.1 μg / ml 18.43 0 1 μg / ml 28.94 0 10 μg / ml 45.92 21.18

*: P <0.01 significantly different from negative control group (student's t -test)

T: Increase in cytotoxicity () = (Experimental cytotoxicity () / Negative control cytotoxicity () × 100)-100

Experimental Example 4

Acute Toxicity Test of Extracts from Combination Preparation

A single dose toxicity test was carried out for each of the extracts of the preparations containing the preparations obtained in Examples 1 to 6 to confirm the safety as follows.

Rat SPF (Specific Pathogen-Free) SD rats (cancer and male) were fed and administered orally to the experimental group (S-1 to S-6) according to the present invention at doses of 1.0, 2.3 and 5.0 g / kg, respectively. As a result of observation for two weeks, 70 cases of hydrated ethanol extract were not observed in all experimental animal groups, and no signs of poisoning and abnormal state were observed. In addition, the findings were not observed might be specific results of the autopsy 2 weeks half lethality (LD ₅₀₎ of each test substance is shown in Table 4 below.

Table 4

Single-dose toxicity test results of extracts from complexes containing manufacture

Experimental group Route of administration Dose (g / kg) Death Clinical symptoms Weight gain Autopsy findings Half lethality (LD _5O ) S-1 Oral administration 1.0 0/5 clear No change clear 〉 5.0 g / kg 2.3 0/5 5.0 0/5 S-2 Oral administration 1.0 0/5 clear No change clear 〉 5.0 g / kg 2.3 0/5 5.0 0/5 S-3 Oral administration 1.0 0/5 clear No change clear 〉 5.0 g / kg 2.3 0/5 5.0 0/5 S-4 Oral administration 1.0 0/5 clear No change clear 〉 5.0 g / kg 2.3 0/5 5.0 0/5 S-5 Oral administration 1.0 0/5 clear No change clear 〉 5.0 g / kg 2.3 0/5 5.0 0/5 S-6 Oral administration 1.0 0/5 clear No change clear 〉 5.0 g / kg 2.3 0/5 5.0 0/5

From the above results, it can be seen that each extract containing the extract of the preparation according to the present invention has no toxicity problem.

The pharmaceutical composition for the prevention and treatment of liver cancer, which is a mixture of the present invention (gumbang), gardenia, gold and shiho in a ratio of 0.7 to 1.5: 1 to 2: 1: 1 to 2, has a novel composition and little toxicity or side effects. In addition, the drug is excellent in terms of the prevention and treatment effect of liver cancer, it is very stable in terms of preparation and has the advantage of obtaining a very useful drug industrially.

Claims (7)

A pharmaceutical composition for the prevention and treatment of liver cancer, containing X, golden, golden, and Xho of Xia. The pharmaceutical composition for preventing and treating liver cancer according to claim 1, wherein the weight ratio of manufactured, gardenia, golden, and shiho is contained in a weight ratio of 0.7 to 1.5: 1 to 2: 1: 1 to 2. The method according to any one of claims 1 to 3, wherein the preparation, gardenia, gold and shiho are 15 to 300 mg containing a weight ratio of 0.7 to 1.5: 1 to 2: 1: 1 to 1 once a few times. Pharmaceutical composition for preventing and treating liver cancer. The pharmaceutical composition for preventing and treating liver cancer according to claim 3, wherein the dosage form of the administrable composition is a tablet, powder, granule, liquid, syrup, capsule. Prepared, Gardenia, Golden and Shiho Herbs were mixed in a weight ratio of 0.7-1.5: 1-1-2: 1: 1-2, refluxed with an organic solvent, extracted and filtered to obtain a filtrate, and the residue was further purified twice with an organic solvent. The filtrate obtained by extraction filtration is combined with the filtrate, concentrated under reduced pressure to prepare an extract, and then formulated into tablets, powders, granules, solutions, syrups, and capsules by containing a pharmaceutically acceptable carrier. A method for producing a pharmaceutical composition for preventing and treating liver cancer, which contains gold and shiho in a weight ratio of 0.7 to 1.5: 1 to 2: 1: 1 to 2. The method of claim 5, wherein the extract is selected from ethanol, methanol or ethyl acetate as an organic solvent. The method of claim 6, wherein the ethanol is 70 saturated ethanol.