KR0156039B1 - Pharmaceutical composition for hepatic disease - Google Patents

Abstract

The present invention removes the fraction obtained by treating with a non-polar solvent in the ethanol extract of the manufactured ethanol extract in order to increase the medicinal effect on the hepatic disease and to solve the toxicity problem, the remaining water fraction and butanol obtained by treating this water fraction with a butanol solvent As fractions were obtained, the water fraction and butanol fraction of the present invention show excellent prophylactic and therapeutic effects against hepatic diseases.

Description

Method for preparing a pharmaceutical extract from the manufacture () and a pharmaceutical composition for the prevention and treatment of liver diseases containing the extract

The present invention relates to a method for preparing a extract having a prophylactic and therapeutic effect on hepatic diseases, and a pharmaceutical composition for the prevention and treatment of hepatic diseases using the extract as an active ingredient.

That is, the present invention is the production of larvae of chafer A method of increasing the effect of preventing and treating hepatic diseases of the manufactured powder, which is used as a medicinal herb, by removing only the fractions with toxicity using a non-polar solvent from the water-containing ethanol extract of) Relates to an extract.

The medicinal herb is an animal medicinal herb found in ancient Chinese medicines, agreements, medicines, etc. Its origin is a larva of insects belonging to the family Scarabeidae, which is used for gout from blood, blood, blood, and anti-inflammatory. It is also used for globules, and in recent years it has been taken as a folk herbal medicine to treat liver disease.

However, in addition to these useful medicaments, they contain high toxicity, which requires special attention (herb wood, Chinese medicine, Dongbobom, medicinal herb), and recently published articles [Acute toxicity of mouse extracts and carbon tetrachloride in rats]. Effect on Hepatic Disorders, Journal of Pharmacognosy, 22 (1), 36-44 (1991)], the hydrous ethanol extracts of the preparations were also effective on hepatic impairment caused by carbon tetrachloride. Toxicities of up to 10% mortality at 1,000 mg / kg have been recognized, and it is necessary to solve these toxic problems in order to use the preparation as a herbal medicine.

Therefore, the present invention is to remove the toxic part of the extract and to extract and separate only the active ingredient in order to develop a prophylactic and therapeutic drug for hepatic diseases using the manufacturing and extracting and separating the pharmacological activity and toxicity of each fraction As a result of comparative study, the preparation was first extracted with hydrous ethanol, and it was confirmed that the toxic component of the preparation could be dissolved and removed in the non-polar solvent. In other words, the water fraction and the n-butanol fraction obtained by re-fractionation of the water fraction and the water fraction and the n-butanol fraction obtained by treating the hydrous ethanol extract with water and a nonpolar solvent were re-fractionated with saturated n-butanol. It appeared much better than the ethanol extract, confirmed that there is no toxicity to complete the present invention.

Hereinafter, the present invention will be described in detail.

The present invention relates to the remaining water fraction obtained by removing a fraction obtained by treating a manufactured hydrous ethanol extract with a nonpolar solvent among various fractions of the prepared extract and an n-butanol fraction obtained by treating the water fraction with saturated n-butanol. Fractions can be used as a prophylactic and therapeutic agent for hepatic diseases.

Suitable non-polar solvents for dissolving and removing toxic substances are n-hexane, ether, petroleum ether, chloroform, dichloromethane, benzene, ethyl acetate, and the like, of which n-hexane is preferably used.

Fractions obtained by treating the ethanol extract prepared in the present invention with a nonpolar solvent exacerbate hepatic function in a dose-dependent manner, or the water fraction remaining after removing the nonpolar solvent fraction has a toxicity problem even in an acute toxicity test using ICR mice. Inhibition of GOT caused by hepatic impairment by 61-96%, GPT 70-95%, and n-butanol fraction had no problem in toxicity, and GOT was 68-94%. By lowering the GPT to 73-96%, it showed a much better effect than the hydrous ethanol extract and solved the toxicity problem at high doses in the hydrous ethanol extract. These effects are superior to diphenyl dimethyl bicarboxylate (DDB), which is currently considered to be the best treatment for hepatic diseases, and has been found to be very useful for the treatment of hepatic diseases.

Water fraction and n-butanol fraction in the extract of the present invention can be used alone or in combination with other herbal formulations, the formulation is a tablet, powder, granules, liquids, syrups, capsules, etc. using a conventional pharmaceutically acceptable carrier Can be prepared by taking.

The water fraction of the manufactured extract of the present invention is preferably divided by the adult adult 20 to 4,000mg amount several times a day, and n-butanol fraction is preferably 5 to 1,000mg the amount divided 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.

Example 1. Isolation of Extract from Preparation:

As a preparation, 500 ml of 70% ethanol was added to 100 g of dried cotyledon (Liocola brevitar sis) oily powder, heated and extracted at 60 ° C. for 3 hours, filtered, and 500 ml of 70% ethanol was added to the residue. After extraction, the filtrates were combined, concentrated under reduced pressure, and freeze-dried to obtain a powdered ethanol extract. Suspended 35g ethanol extract was suspended in 500ml of distilled water, and then n-hexane was added to eastern blood. Obtain two fractions, a hexane layer and a water layer. The same amount of hexane was added again to the water layer fraction, and the separation process of obtaining the hexane layer fraction was carried out two more times. The hexane layer fractions were combined, concentrated under reduced pressure, lyophilized to obtain an n-hexane fraction, and the yield was 7 g. After concentration under reduced pressure, a water fraction of the preparation was obtained, and this was expressed as water fraction 1 solid content of 28 g. The water layer from which the n-hexane fraction obtained above was removed was fractionated into an n-butanol water layer using a saturated n-butanol of eastern blood. This operation was repeated four more times to combine all the n-butanol layers, and concentrated under reduced pressure to obtain an n-butanol fraction (hereinafter referred to as butanol fraction). The yield 5.5g and the remaining water layer were also concentrated under reduced pressure to obtain a water fraction 2. A summary of the process of yield 22.5g or more is as follows.

[Experiment 1. Comparative experiment of pharmacological effect of each fraction of manufactured extract:]

In order to confirm the effect of water-soluble ethanol extract of rats treated with galactosamine, SD male male rats were purchased from Charles River (Japan), and treated with ① galactosamine alone and ② test substance and galactosamine-treated groups. Each group was divided into 10 groups and intraperitoneally injected 400 mg / kg of galactosamine dissolved in distilled water for injection to cause acute hepatitis.

The hydrated ethanol extract of the preparation was orally administered 24 hours and 1 hour before galactosamine administration, and 24 hours after galactosamine administration, blood was collected from the abdominal aorta by ether under anesthesia, and coagulated at room temperature for 1 hour. The serum GOT and GPT values were measured by blood biochemical analyzer (DIMENSION, Dupont, USA).

The aqueous ethanol extract of the preparation obtained in Example 1 was suspended in 0.5% carboxymethyl cellulose solution to prepare a daily dosage of 50, 200, 800 mg / kg body weight based on the extract and orally administered to the control material. Phosphoric didibi (diphenyl dimethyl bicarboxylate (DDB)) was also suspended in 0.5% carboxymethyl cellulose solution, and the daily dose was prepared orally so that the daily dose would be 50 mg / kg based on DDB. At the same time, the hexane fraction and the water fraction 1 obtained in Example 1 were tested in the same manner to confirm the therapeutic effect in the galactosamine-induced acute liver injury model. The doses of the two test substances to the rats were determined according to the extraction ratio (hexane fraction: water fraction 1 = 1: 4) of the two fractions obtained by solvent fractionation of the hydrous ethanol extract of the preparation. Suspended in 0.5% carboxylic acid cellulose solution, and orally administered. The experimental results are shown in Table 1.

From the results of Table 1, it could be seen that the 50, 200 mg / kg dose of the hydrated ethanol extract of the preparation showed a prophylactic and therapeutic effect of hepatic disease. However, at the 800 mg / kg dose, the medicinal effect decreased because of hexane. The cause can be seen from the experimental results for the administration group of fractions. That is, since the hexane fraction exacerbates liver failure in a dose-dependent manner, it can be seen that toxic fractions are included in the hexane fraction. Therefore, the hydrated ethanol extract containing the hexane fraction is more toxic at higher doses. The efficacy of water fraction 1 from which the hexane fraction was removed showed an increase in the effective rate of about 10% in the low dose range compared to the efficacy of the hydrous ethanol fraction. It showed a prophylactic and therapeutic effect of hepatic disease, which shows a significant drug.

* (P0.05), ** (P0.01) means that the student's t-test showed a significant difference compared to the galactosamine control group.

The effective rate (%) in Table 1 is (control-test group / control-normal group) × 100.

[Experimental Example 2 Comparative Experiment of the Effect of Butanol Fraction and Water Fraction 2 in the Extracts Prepared:]

In order to compare the therapeutic effects in the acute liver injury model caused by galactosamine of water fraction 2 and butanol fraction, which are two new fractions obtained by continuing fractionation from water fraction 1 obtained in Example 1, The same procedure was followed to obtain the result shown in Table 2. The doses of the two test substances to the rats were determined according to the extraction ratio (butanol fraction: water fraction 2 = 1: 4) of the two fractions extracted from the water fraction 1, and the two test substances were prepared in the same manner as in Experimental Example 1. By oral administration. The test results are shown in Table 2 below.

* (P0.05), ** (P0.01) means that the student's t-test showed a significant difference compared to the galactosamine control group.

In the results of Table 2, butanol fraction was superior in the dose-dependent prevention and treatment of hepatic disease, and showed a 10 to 20% increase in drug efficacy compared to the same dose of water fraction 1 When compared with the same dose, about 20% of drug efficacy was confirmed, indicating that it is useful as an agent for preventing and treating liver disease.

[Experimental Example 3. Acute toxicity comparison of each fraction of the extract extract:]

Acute toxicity tests were performed on the water fraction 1, the hexane fraction, and the butanol fraction obtained in Example 1 to confirm the safety of each fraction as follows.

Water fractions and butanol fractions were observed in two animals after 1 week of oral administration of water fraction 1, butanol and hexane fractions at 0.63, 1.25, 2.5 and 5 g / kg doses in male ICR mice. No specific poisoning symptoms or general abnormalities were observed, but hexane fractions were observed. After two weeks of autopsy, no unusual findings were observed, and the half-lethality (LD) of each test substance is shown in Table 3 below.

In addition, the male ICR mice were intraperitoneally administered at 0.13, 0.25, 0.5, and 1.0 g / kg doses for 7 days. No mortality was observed in the water fraction 1 and butanol fraction. Death was observed. After 7 days of administration of the test substance, no significant findings were observed in the water fraction 1 and butanol groups. However, in the hexane fraction group, intra-abdominal organ adhesion was observed. These adhesions are concentration dependent, and severe adhesions of the stomach, duodenum and pancreas and liver were found in all animals of the 1 g / kg administration group. In the case of intraperitoneal administration, the half lethality (LD) of each test substance is shown in Table 3 below.

As a result, it was found that the hexane fraction is required to be removed from the hydrous ethanol extract of the production because the toxicity is recognized, water fraction 1 and butanol fraction is no problem in toxicity.

Composition Example 1. Granules (value in weight%):

Butanol Fraction Example 1 5

Lactose 68

Starch 21

Hydroxypropyl Cellulose 5

Magnesium Stearate 1

The ingredients were mixed and kneaded using ethanol corresponding to about 30% by weight of the total mixture as an emollient, then granulated and dried to prepare granules containing 50 mg of dried butanol fraction in 1 g each.

[Composition example 2. Refining:]

The granules prepared in Composition Example 1 were compression molded by 500 mg per tablet to prepare a tablet containing 25 mg of butanol.

Composition 3 Syrup (Weight%):

Water fractionation example 1 5

50 per bag

Methylparaben 0.08

Propylparaben 0.02

Spices 0.1

Correction quantity

After heating and dissolving sucrose, methyl paraben and propyl paraben in a predetermined amount of water, add a water fraction to make a homogeneous solution, and mix by adding flavoring rule.

The final dose was adjusted to 100ml, filtered and the filtrate was filled in 30ml bottles in a conventional manner and sterilized to prepare a syrup containing 1,500 mg of water fraction 1 per bottle.

Claims (4)

A method of preparing an extract of the preparation by extracting the preparation with brine ethanol and extracting the extract extract with a nonpolar solvent and water to remove the nonpolar solvent fraction and concentrated to dryness. The process according to claim 1, wherein the nonpolar solvent is n-hexane, ether, petroleum ether, chloroform, dichloromethane, benzene or ethyl acetate. The extract obtained by dissolving the prepared extract obtained in the water fraction of claim 1 in water, extracting it with n-butanol or saturated n-butanol solvent, and removing the water fraction from the fraction obtained by concentrating and drying the obtained saturated n-butanol fraction. How to prepare. A pharmaceutical composition for the prophylaxis and treatment of hepatic diseases, comprising as an active ingredient a manufactured extract prepared by the method of any one of claims 1 to 3.