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

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

Inactive Publication Date: 2011-05-18
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 activity of Euphorbia euphorbia, the root plant of the traditional Chinese medicine Euphorbia euphorbia. It is known that its roots contain a small amount of diterpenes, triterpenes and other components (Kong Lingyi, etc., 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
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1 Extraction and separation method of diterpene compound isoeuphpekinensin

[0031] One kilogram of dried roots of Euphorbia euphorbia growing in the suburbs of Nanjing, Jiangsu, China were selected and extracted with 2.5 times the amount (volume) of 95% ethanol for 3 times by heating and refluxing. The 3 extractions were combined and the ethanol was recovered under reduced pressure to obtain 68.3 g of extract; The liquid extract was chromatographed on a silica gel column (200-300 mesh) and eluted with petroleum ether and petroleum ether: ethyl acetate gradient (100:0, 99:1, 98:2, 96:4, 80:20) , Thin-layer chromatography detection, collect the fractions containing isoeuphpekinensin, combine and concentrate, then subject to silica gel column chromatography, eluting with petroleum ether-ethyl acetate (100:0, 99:1, 98:2) gradient, thin layer Chromatographic detection, the pure diterpene compound isoeuphpekinensin was obtained.

Embodiment 2

[0032] Example 2 In vitro anti-tumor test (Alamar blue method)-human lung cancer cell line NCI-H1299

[0033] The human lung cancer cell line NCI-H1299 uses RPMI-1640 medium containing 10% calf serum, 100U / ml penicillin, 0.1mg / ml streptomycin at 37℃, 5% CO 2 Routine cultivation in an incubator. It was digested and passaged with 0.25% pancreatin plus 0.02% EDTA. Take the logarithmic growth phase cells, digest by trypsin, prepare a cell suspension with RPMI-1640 medium containing 10% calf serum, the cell concentration is about 1×10 5 Pcs / ml, inoculated in a 96-well culture plate, 180μL per well; in each well of the experimental group, add 20μL of a solution containing samples of different concentrations of isoeuphpekinensin (samples are dissolved in dimethyl sulfoxide, medium explanation), and 20μL of dimethyl sulfoxide Sulfone (final concentration 2 After 48 hours of incubation in the incubator, add 20μl of Alamarblue solution to each well, continue to incubate for 4 hours, use Sp...

Embodiment 3

[0034] Example 3 In vitro anti-tumor test (Alamar blue method)-human nasopharyngeal carcinoma cell line KB

[0035] The specific method is the same as in Example 2, the sample concentration is 2.5, 5, 10, 12.5, 25 μg / ml, and the inhibition rate is 3.35%, 24.91%, 48.35%, 79.73%, 93.54%. Then use SPSS software to calculate the IC of isoeuphpekinensin inhibiting KB tumor cell line 50 It is 8.52μg / 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 following 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 medicine, in particular to an anti-tumor diterpene compound isoeuphpekinensin extracted and isolated from the plant Euphorbia root and an extraction method of the compound. Background technique [0002] Euphorbia pekinensis Rupr (Euphorbia pekinensis Rupr) is a plant belonging to the Euphorbia family Euphorbia. Its root is called Beijing Euphorbia. It is a traditional Chinese medicine in my country. It is included in the previous editions of "The Pharmacopoeia of the People's Republic of China". The effect of dispelling lumps is often used in modern clinical practice for cirrhosis and ascites and pleural effusion caused by various reasons. Euphorbia is mainly produced in Nanjing, Yangzhou, Pizhou and other places in Jiangsu. In addition, it is also produced in Sichuan, Fujian, Jiangxi, Hunan, Hubei and other places. Although there are many studies on the chemical components and pharmacologi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
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