CN104341430B - A kind of soil Radix Glycyrrhizae A and extracting method thereof and purposes - Google Patents

Abstract

The invention discloses a soil licorice A and its extraction method and application. The structural formula of the soil licorice A is: The chemical name of this licorice A is 3-(p-methoxyphenyl)-4-hydroxy-5-methoxy-2', 2'-dimethyl-6, 7-dihydropyranocoumarin, The molecular formula is C ₂₂ H ₂₀ O ₆ , and the relative molecular weight is 380. The soil licorice A of the present invention has biological activities such as anti-tumor, and its extraction method is simple and easy, and the extraction rate is high, which can be effectively applied to the preparation of anti-tumor drugs, and can effectively treat human gastric cancer cells, liver cancer cell lines, and human lung cancer cells. , cervical cancer cells, ovarian cancer cells, etc. have a strong inhibitory effect.

Description

A kind of soil licorice A and its extraction method and application

technical field

The invention relates to the technical field of medicinal chemistry, in particular to a soil licorice A and an extraction method thereof, and the use of soil licorice A to prepare antitumor drugs.

Background technique

Cancer has risen to become the second largest "killer" disease of human beings after cardiovascular disease. Cancer and cardiovascular disease have become the leading causes of death. The prospect of finding and developing effective new anti-tumor drugs from plants is broad. At present, many natural active products with different structures and mechanisms of action have become a hot spot in the research of new anti-tumor drugs at home and abroad. Among the two batches of major new drug development projects launched continuously in my country, the research and development of plant-derived drugs, especially natural drugs using plant resources in my country, has been listed as key support objects. Due to the unique mechanism of action of plant-derived anti-tumor drugs, the anti-cancer efficacy Significantly, it has gradually occupied a dominant position in clinical application.

According to the records of "Flora of China", there are 28 species of Dalbergia plants (Dalbergia) in my country, and only two species have been studied at present, especially Dalbergia odorifera has been studied the most. Many pharmacological activities have shown that balsamic has anti-diabetic, anti-oxidant, anti-hyperglycemic, anti-inflammatory, tumor and antibacterial effects, so it is very meaningful to develop diabetes drugs from this genus. Dalbergia benthami Prain, also known as: banana vine hemp, Liangyue sandalwood, vines, grows in sparse forests and bushes, and is distributed in Guangdong, Guangxi, Hainan and other places. The medicinal part is the stem, which can be used to promote menstruation and activate blood circulation, and mainly treat symptoms such as bruises, muscle and bone pain, and irregular menstruation caused by qi stagnation and blood stasis.

At present, there is no literature report on the study of this plant. The present invention isolates and identifies a compound from the plant for the first time, which is identified as the soil licorice A by NMR and mass spectrometry, and the content in the plant is as high as 3.5%. The compound was originally derived from Derris eriocarpa How, but the content in this plant is low, which limits its application. Soil licorice A belongs to the coumarin compound, which has anti-tumor and other biological activities. Therefore, this patent provides a strong guarantee for the development and application of this compound.

Contents of the invention

One object of the present invention is to provide a soil licorice A, which has biological activities such as antitumor. Another purpose is to provide a method for extracting the licorice root A, and the licorice root A can be used in the preparation of anti-tumor drugs, so that the extracted anti-tumor drugs can effectively prevent and inhibit tumor progression.

In order to solve the above technical problems, the present invention provides licorice root A and its extraction method and use.

The structural formula of Soil Glycyrrhizae A of the present invention is:

The chemical name of this licorice A is: 3-(p-methoxyphenyl)-4-hydroxyl-5-methoxy-2',2'-dimethyl-6,7-dihydropyranocoumarin ;

The molecular formula of the soil licorice A is: C ₂₂ H2 ₀ O ₆ ;

The relative molecular weight of the soil licorice A is: 380.

Can be extracted to this licorice root A by following method, it comprises the steps:

1) The coarse powder of Dalbergia japonica is used as a raw material, and ethanol is used as a solvent, and the percolation method is used for extraction, and the extract extract is obtained after evaporating the solvent.

2) add water to suspend the extract extract, then add petroleum ether to the suspension to extract and separate the extract from petroleum ether, then add ethyl acetate to the suspension to extract to obtain the extract from ethyl acetate;

3) Take the extract from the ethyl acetate part, and repeatedly separate it by silica gel column chromatography to obtain a colorless and transparent granular crystalline compound, which is the target product, Licorice Root A.

The licorice root A can be used to prepare antitumor drugs, for example, to prepare one or more drugs in antitumor injections, tablets, pills, capsules, suspensions or emulsions.

Beneficial effects of the present invention: the soil licorice A extracted from Dalbergia japonica in the present invention has biological activities such as anti-tumor. Moreover, the extraction method of the present invention is simple and easy to implement, has a high extraction rate, can be effectively applied to the preparation of antineoplastic drugs, and can be effective against human gastric cancer cells, liver cancer cell lines, human lung cancer cells, cervical cancer cells, ovarian cancer cells, etc. Strong inhibitory effect.

detailed description

The present invention will be further described in detail below in conjunction with the embodiments, so that those skilled in the art can implement it with reference to the description.

Example 1

The extraction method of soil licorice A of the present invention comprises following specific steps:

1) Take 10.0kg of the coarse powder of Dalbergia japonica, extract by percolation with 10 times the amount of 80% ethanol, combine the extracts, and evaporate the solvent to obtain the extract extract;

2) add water to suspend the extract extract, then add petroleum ether to the suspension to extract and separate the extract from petroleum ether, then add ethyl acetate to the suspension to extract to obtain the extract from ethyl acetate;

3) Take 30 g of the extract from the part of ethyl acetate, and repeatedly separate it by silica gel column chromatography to obtain a colorless and transparent granular crystalline compound, which is the target product Glycyrrhizae A. The way of this silica gel column chromatographic separation can be according to following operation: first with chloroform-acetone gradient elution (chloroform → 80: 1 → 50: 1 → 30: 1 → 15 _: 1 → 7: 1 → 3: 1 → acetone), A total of 358 fractions were collected, combined with TLC detection, and the same fractions were combined to obtain 14 components (A, B, C, D, E, F, G, H, I, G, K, L, M, N); wherein, component I is a white powder, then take 300mg component I, and elute with petroleum ether-acetone gradient (15:1 → 7:1 → 3:1), collect 103 fractions altogether, use TLC The detection and identification were combined, and the same fractions were combined. The obtained components were eluted in petroleum ether-acetone (7:1), and 80.55 mg of a colorless and transparent granular crystalline compound was obtained. The colorless and transparent granular crystalline compound was Glycyrrhizae A .

The structural formula of the Radix Glycyrrhizae A that above-mentioned embodiment 1 extracts is:

The chemical name of this licorice A is 3-(p-methoxyphenyl)-4-hydroxy-5-methoxy-2',2'-dimethyl-6,7-dihydropyranocoumarin, The molecular formula is C ₂₂ H ₂₀ O ₆ , and the relative molecular weight is 380. The physical and chemical properties obtained by the physical and chemical analyzer are: colorless transparent granular, mp203-206 °C. EI-MS(m/z): 381[M+1] ⁺ , 380[M] ⁺ , 315[M-15], 337, 321, 307, 233, 232, 217, 203, 175, 161, 148, 138, 133. ¹ H-NMR (400MHz, CDCl ₃ ) δ: 9.97 (1H, s, 4-OH), 7.46 (2H, dd, J=11.4Hz, 2.9Hz, H-2′, H-6′), 6.97( 2H, dd, J=8.7Hz, 2.9Hz, H-3′, H-5′), 6.63 (1H, s, H-Ar), 6.50 (1H, d, J=10Hz, H-4″), 5.77 (1H, d, J=10Hz, H-3″), 3.96 (3H, s, 4′-OCH ₃ ), 3.77 (3H, s, 5-OCH ₃ ), 1.48 (6H, s, -CH ₃ ). ¹³ C-NMR (125MHz, CDCl ₃ ) δ: 160.1 (C-2), 101.7 (C-3), 162.6 (C-4), 153.7 (C-5), 124.2 (C-6), 157.6 (C -7), 110.6(C-8), 152.1(C-9), 103.8(C-10), 123.2(C-1′), 131.4(C-2′), 113.5(C-3′), 158.8 (C-4'), 113.5(C-5'), 131.4(C-6'), 77.5(C-2"), 115.0(C-3"), 131.7(C-4"), 64.4(3 '-OCH ₃ ), 55.2 (5-OCH ₃ ).

Example 2

The soil licorice A that is extracted from the above-mentioned embodiment 1 is carried out in vitro anti-tumor activity experiment:

MTT method is widely used in the activity detection of some biologically active factors, large-scale antitumor drug screening, cytotoxicity test and tumor radiosensitivity determination, etc. It has the characteristics of high sensitivity and economy. Exogenous MTT can be reduced to water-insoluble blue-purple crystalline formazan (Formazan) by succinate dehydrogenase in the mitochondria of living cells and deposited in the cells, and its light absorption value is measured at a wavelength of 490nm by an enzyme-linked immunosorbent assay , which can indirectly reflect the number of living cells. Within a certain cell number range, the amount of MTT crystal formation is proportional to the cell number. By calculating the concentration of the corresponding drug when the inhibitory rate of the compound on tumor cells is 50%, it is called the half inhibitory concentration IC ₅₀ , that is, the half inhibitory concentration. Generally speaking, the smaller the value of IC ₅₀ , it indicates that the drug inhibits the cells. The higher the activity. In this experiment, the Bliss method was used to calculate the IC ₅₀ value of licorice A and 5-FU of the present invention on human gastric cancer cells, liver cancer cell lines, human lung cancer cells, cervical cancer cells, ovarian cancer cells, etc. when the inhibition rate was 50%. . The experimental results are shown in the table below:

From the results of Example 2, it can be seen that the soil licorice A of the present invention has a strong anti-tumor active component through anti-tumor experiments in vitro. The licorice root A of the present invention can be made into one or more medicines in antitumor injections, tablets, pills, capsules, suspensions or emulsions. The licorice root A of the present invention and the medicine extracted therefrom can exert a strong inhibitory effect on human gastric cancer cells, liver cancer cell lines, human lung cancer cells, cervical cancer cells, ovarian cancer cells and the like. From the comparative data in the table, it can be seen that its inhibitory effect on cervical cancer cells HeLa is the most obvious, and its therapeutic effect is better, and the inhibitory effect of Glycyrrhizae A of the present invention on other cancer cells is also better than that of 5-FU. The invention provides a new idea for researching and developing new antitumor drugs.

Although the embodiment of the present invention has been disclosed as above, it is not limited to the use listed in the specification and implementation, it can be applied to various fields suitable for the present invention, and it can be easily understood by those skilled in the art Therefore, the invention is not limited to the specific details without departing from the general concept defined by the claims and their equivalents.

Claims (1)

1. an extracting method of Glycyrrhizae A, is characterized in that, comprises the following steps to extract: 1) using the coarse powder of Dalbergia japonica as a raw material, using ethanol as a solvent, extracting by percolation, and obtaining the extract extract after evaporating the solvent; 2) add water to suspend the extract extract, then add petroleum ether to the suspension to extract and separate the extract from petroleum ether, then add ethyl acetate to the suspension to extract to obtain the extract from ethyl acetate; 3) Take the extract from the ethyl acetate part, and repeatedly separate it by silica gel column chromatography to obtain a colorless and transparent granular crystalline compound, which is the target product, Licorice Root A.