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Anti-tumor diterpenoid isoeuphpekinensin in euphorbia perkinensis root and extraction method thereof

A compound and anti-tumor technology, applied in the separation/purification of anti-tumor drugs, carbonyl compounds, organic chemistry, etc., can solve the problems of few research reports on chemical composition and biological activity

Inactive Publication Date: 2009-04-01
NANJING UNIVERSITY OF TRADITIONAL CHINESE MEDICINE +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although there are many studies on the chemical constituents and pharmacological activities of Euphorbia plants at home and abroad, the hot report is that Euphorbia plants are rich in strong anti-tumor diterpene components (Shi Yanping, etc. Natural Product Research and Development, 1998, 11( 5): 85-90), but there are few reports on the chemical constituents and biological activities of Euphorbia euphorbia, the root plant of the traditional Chinese medicine Euphorbia euphorbia, and it is known that its roots contain a small amount of diterpenes, triterpenes and other components (Kong Lingyi et al., Acta Pharmaceutica Sinica, 1996, 31(7): 524-527; Kong LY, et al. Planta Medica, 2002, 68(3), 249-252)

Method used

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  • Anti-tumor diterpenoid isoeuphpekinensin in euphorbia perkinensis root and extraction method thereof
  • Anti-tumor diterpenoid isoeuphpekinensin in euphorbia perkinensis root and extraction method thereof
  • Anti-tumor diterpenoid isoeuphpekinensin in euphorbia perkinensis root and extraction method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0031] Example 1 Extraction and separation method of diterpenoid isoeuphpekinensin

[0032] Select 1 kilogram of dried roots of Euphorbia sp. grown in the suburbs of Nanjing, Jiangsu, China, and extract 3 times with 2.5 times the amount (volume) of 95% ethanol under heating and refluxing, combine the 3 extractions, and recover ethanol under reduced pressure to obtain 68.3 grams of liquid extract; The stream extract was subjected to silica gel column (200-300 mesh) chromatography, eluted with petroleum ether and petroleum ether: ethyl acetate gradient (100:0, 99:1, 98:2, 96:4, 80:20) , detected by thin layer chromatography, collected the fractions containing isoeuphpekinensin, combined and concentrated, and then subjected to silica gel column chromatography, eluted with petroleum ether-ethyl acetate (100:0, 99:1, 98:2) gradient, thin layer Chromatographic detection, the diterpene compound isoeuphpekinensin pure product.

Embodiment 2

[0033] Example 2 In vitro anti-tumor test (Alamar blue method)-human lung cancer cell line NCI-H1299

[0034] Human lung cancer cell line NCI-H1299 was cultured in RPMI-1640 medium containing 10% calf serum, 100U / ml penicillin, and 0.1mg / ml streptomycin at 37°C, 5% CO. 2 Routine culture in an incubator. Digested with 0.25% trypsin plus 0.02% EDTA and passaged. Take cells in logarithmic growth phase, digest with trypsin and prepare cell suspension with RPMI-1640 medium containing 10% calf serum, the cell concentration is about 1×10 5 1 / ml, inoculated in a 96-well culture plate, 180 μL per well; 20 μL of solutions containing isoeuphpekinensin samples with different concentrations were added to each well of the experimental group (samples were dissolved in dimethyl sulfoxide, explained in the medium), and 20 μL of dimethyl sulfoxide was added to each well. Sulfone (final concentration 2 After culturing in the incubator for 48 hours, 20 μl of Alamarblue solution was added to eac...

Embodiment 3

[0035] Example 3 In vitro antitumor test (Alamar blue method) - human nasopharyngeal carcinoma cell line KB

[0036] The specific method is the same as that in Example 2, the sample concentrations are 2.5, 5, 10, 12.5, and 25 μg / ml, respectively, and the inhibition rates are 3.35%, 24.91%, 48.35%, 79.73%, and 93.54%, respectively. SPSS software was used to calculate the IC of isoeuphpekinensin inhibiting KB tumor cell line 50 was 8.52 μg / ml.

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Abstract

The invention discloses an anti-tumor diterpenoid compound isoeuphpekinensin which is separated from euphorbia roots, the chemical structure is as the right formula, the chemical structure and the spatial configuration of the compound are determined via the spectral analysis, the integrated analysis of nuclear magnetic resonance spectroscopy and the X-ray single crystal diffraction technology particularly. The preparation method is as follows: the fresh euphorbia root is cut into slices and dried, then ethanol is used for reflux extraction, the ethanol is recycled under reduced pressure, a liquid extract is obtained, and then the chromatographic separation is carried out on the liquid extract by a silica gel column, thereby obtaining the compound. In vitro anti-tumor experiments show that the compound has significant inhibition effect on human nasopharyngeal carcinomas cells, KB cells and others which are six types of tumor cell lines in total. The invention provides the lead compound for developing new anti-tumor medicaments, thereby having important significance on developing and utilizing medicinal plant resources of China.

Description

technical field [0001] The invention relates to the technical field of traditional Chinese medicines, in particular to an antitumor diterpene compound isoeuphpekinensin extracted and isolated from the root of Euphorbia plant and an extraction method of the compound. Background technique [0002] Euphorbia pekinensis Rupr (Euphorbia pekinensis Rupr) is a plant of the Euphorbiaceae family. Its root is called Jingjingji and used as medicine. It is a traditional Chinese medicine in my country. The efficacy of Sanjie is commonly used in modern clinical practice for liver cirrhosis ascites and pleural effusion caused by various reasons. Euphorbia is mainly produced in Nanjing, Yangzhou, Pizhou and other places in Jiangsu, in addition, Sichuan, Fujian, Jiangxi, Hunan, Hubei and other places are also produced. Although there are many studies on the chemical constituents and pharmacological activities of Euphorbia at home and abroad, it is hotly reported that Euphorbia is rich in dit...

Claims

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Application Information

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IPC IPC(8): C07C47/36C07C45/78A61K31/11A61P35/00A61K36/47
Inventor 梁侨丽戴传超蒋继宏
Owner NANJING UNIVERSITY OF TRADITIONAL CHINESE MEDICINE
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