JPH0253730A - Collection of antitumor substance - Google Patents

Abstract

PURPOSE:To collect an antitumor substance from TETSUJUYOU, Xanthii fructus, TAKUSHITSU and Dianthi herba of crude drug. CONSTITUTION:TETSUJUYOU, Xanthii fructus and TAKUSHITSU are extracted with an acidic lower alcohol and the solvent is distilled away to give an extracted essence. The essence is dissolved in a dilute aqueous solution of alkali, shaken with n-butanol, the butanol layer is collected and butanol is distilled away to give an antitumor substance. An amount of acid added to an acidic lower alcohol solution is varied depending upon crude drugs and is generally <=1%. Extraction is carried out under heating or at room temperature and the alkali concentration of the aqueous solution of alkali is <=1%. To extract an antitumor substance from Dianthi herba, Dianthi herba is firstly treated with a lower alcohol, the alcohol extracted solution is extracted with hot water and water is distilled away from the hot water extracted solution under reduced pressure to give an extract.

Description

DETAILED DESCRIPTION OF THE INVENTION [Industrial Application Field] The present invention relates to a method for collecting antitumor substances from crude drugs, iron tree leaves, soybean grass, lacquer lacquer, and soybean grass.

[Conventional technology]

The development of antitumor agents that are effective for the human body is an important medical issue, and the antitumor activity of many substances has been tested. These substances can be broadly classified into synthetic substances and naturally extracted substances, and herbal medicines are one of the raw materials for naturally extracted substances.

Each Chinese herbal medicine has its own unique pharmacological action, and the Middle Ages Dictionary (edited by Jiangsu New Medical Academy, 1977, published by Kamiyu People's Publishing House) lists more than 6,000 kinds of herbal medicines, and they are said to have antitumor effects. There are more than 100 different types. For about half of these, the actual antitumor activity has been elucidated, but for the rest, the actual antitumor activity has not been elucidated. However, even though the active ingredients are unknown, it is thought that there are signs of relatively good anticancer activity, and there are many Chinese herbal medicines that are combined with other herbal medicines and used as anticancer drugs. In Chinese medicine, only a single herbal medicine is rarely used, but a large number of herbal medicines are mixed together and a decoction is used. Therefore, the result is determined as the integrated effect of the water-soluble components of the mixed herbal medicines, and the mixed component side The effects of each are not clear.

Therefore, it is unclear which of the herbal medicines used as anticancer drugs contain antitumor substances and whether they are efficiently extracted into decoctions. Tetsuju leaf, Somigusa, Sawa lacquer, and Somugi are each thought to have anti-cancer, anti-tumor, and passion diuretic effects when used alone, and are known as Fujikoto, Sanmoto, and Nisento.

It is used as a Chinese herbal medicine in the treatment of various cancers. However, it is unclear whether these herbal medicines contain antitumor substances.

(Problems to be Solved by the Invention) The purpose of the present invention is to confirm that the anti-tumor substances are contained in the iron tree leaf 2-sang ear grass, the lacquer lacquer, and the barley, and to provide a method for collecting the same.

(Means and effects for solving the problem) As a result of examining the methods for collecting antitumor substances from iron tree leaves, somia grass, sawa lacquer, and konmugi, it was found that all of these herbal medicines contain strong antitumor substances. It is recognized that anti-tumor substances can be collected using the same collection method from iron tree leaves, Somia grass, and Sawa lacquer, but anti-tumor substances cannot be collected from Soumai using the above-mentioned three methods. Knew. Extract iron tree leaves, blue ear grass, and sawa lacquer with acidic lower alcohol, and remove the solvent to obtain the extract. This extracted extract was dissolved in a dilute alkaline aqueous solution, and
- After adding butanol and shaking, collect the butanol layer and add butanol to obtain the antitumor substance. This substance can be purified and purified by chromatography or other conventional methods. The amount of acid added to the acidic lower alcohol solution to be used varies depending on the type of herbal medicine, but it is generally 1% or less, preferably 0.3 to 0.3%, based on the lower alcohol.
The amount is about 0.5%, and the lower alcohol is preferably methanol or ethanol. Extraction is heated.

This can be done at room temperature. The alkaline concentration of the alkaline aqueous solution is 1% or less, preferably about 0.1%.

In order to collect anti-tumor substances from the barley, the barley is first treated with lower alcohol, the alcohol extraction residue is extracted with hot water, and the hot water extract is mixed with water under reduced pressure to obtain an extract. The extract is used as an antitumor substance. If necessary, this extract is further dissolved in a weak acid or a weak alkali, and extracted with n-butanol to collect a butanol extract. This operation further purifies the sample. It is also possible to purify by chromatography or other methods.

The substance thus obtained is stable and has strong antitumor activity as shown below, and can be used alone or in combination with other drugs as an anticancer agent.

Considering the extraction method used and the nature of the material collected,
The anti-tumor substances obtained from iron tree leaves, blue ear grass and sawa lacquer contain nitrogen-containing compounds and alkaloids? It is thought that the substance obtained from R wheat contains phenolic compounds or glycosides as a main component, and the antitumor properties of these harvested products are thought to be due to their alkaloids, phenolic properties, or glycosides.

The present invention will be specifically explained below with reference to Examples.

Example 1 0.2j2 of methanol was added to 22.2 g of Blue Ear Grass, 5-concentrated hydrochloric acid was added thereto, and the mixture was extracted by heating under reflux for 1 hour. This operation was repeated twice, the extracts were combined, and the solvent was distilled off under reduced pressure to obtain an extracted extract.

This extracted extract was added to 550 d of 0.1% caustic soda aqueous solution.
The butanol solutions were combined and the butanol was distilled off under reduced pressure to obtain a solid 1.
I gained 17g.

Analysis of this solid by ultraviolet absorption spectrum revealed that it had a maximum absorption at 218+wμ in methanol, and shoulders at 320 and 400 mμ.

Example 2゜Example 1 except that 22.2g of iron tree leaves were used in place of the blue ear grass. The same operation as above was performed to obtain 3.52 g of solid. As a result of ultraviolet absorption spectrum analysis of this solid, it has maximum absorption at 228 and 282 mμ in methanol.

Example 3 Example 1 except that 22.2g of ゜Sawa urushi was used instead of Soumiso
．． The same operation as above was performed to obtain 36.9 g of solid. As a result of analyzing this solid by ultraviolet absorption spectrum, it was found that 2
It has maximum absorption at 15 and 400 mμ and a shoulder at 26 mμ.

Example 4 0.1!1 M of methanol was added to 24.4 g of ゜mugi and extracted by heating under reflux for 1 hour. Methanol is filtered and residual medicine is 0.21! Add water and heat under reflux for 1 hour for extraction. The solvent was distilled off under reduced pressure to obtain 1.08 g of solid. As a result of analyzing this aqueous solution by ultraviolet absorption spectrum, 2
It has maximum absorption at 12.269 and 331 mμ.

Example 5 Dissolve 4g of ゜mugi in 400rd purified water and adjust the pH with dilute hydrochloric acid.
The mixture was extracted with 100-n-butanol four times, and the butanol extracts were combined and the solvent was distilled off under reduced pressure to obtain 786 mg of the active substance. Note that no activity was observed in the aqueous layer.

Example 6 In vitro cell killing effect on cultured cells (L-5178Y) derived from mouse leukemia.

The active substance collected according to the present invention is dissolved in physiological saline and leukemia cells L-5178Y1, OX are subcultured in RP old-1640 medium containing 10% tallow serum.
After adding 10' cells/ml and culturing in a 5% carbon dioxide incubator at 37°C for 48 hours, the living cells of the control group and the sample addition group were examined using trypan blue staining to determine the presence of each active substance. 50% inhibitory concentration pivot (IC5
゜) was calculated. The results are shown in Table 1.

Table 1. Tumoricidal cell effect on L-5178Y in vitro
, P-, which has been passaged intraperitoneally once a week in mice.
388 ascites cancer cells 1. 106 OX/mouse were implanted intraperitoneally, and the active substance of the present invention was intraperitoneally administered 24 hours after implantation. The frequency of administration was once a day for 5 days, and the survival rate was calculated by comparing the survival days with that of the untreated group. The results are shown in Table-2.

The rate was calculated. The results are shown in Table 3.

Table 2, Antitumor activity against experimental animal tumor P-388 Sample Dose Life extension rate Iron tree leaf 2
00mg/kg x 5 times 11.1% Soumiagusa
200 // X5〃18.1% Sawa lacquer 200 //
X 5 // 10.2% barley 200 // X
5 // 20.1%Table 3. Antitumor effect on experimental animal tumor Sarcoma 180 Sample dose prolongation survival rate Soomi Kusazawa Lacquer Iron Tree Leaves 100mg/kg x 5, F 100mg/kg x 5 tt 100mg/kg x 5 times 100mg/kg x 5 〃 Example 8゜Animal experiment Antitumor test using Sarcoma 180

Claims (2)

[Claims] (1) A method for collecting an anti-tumor substance characterized by dissolving an acidic alcohol extracted substance of iron tree leaves, blue ear grass, or sawa lacquer in an alkaline aqueous solution, extracting the alkaline aqueous solution with butanol, and collecting a butanol extract. . (2) A method for collecting an anti-tumor substance, which comprises collecting a hot water extract of the residue of lower alcohol extraction of porgy.