Method for fast screening topoisomerase I inhibitor from natural product

A technology of topoisomerase and natural products, which is applied in the field of natural product analysis, can solve the problems of not being able to provide detailed features of protein-ligand interactions, false negatives, etc., and achieve fast sample analysis, strong specificity, less effect

Active Publication Date: 2016-06-29
WUHAN BOTANICAL GARDEN CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, spectroscopic methods are prone to false negative and false positive results due to background interference;

Method used

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  • Method for fast screening topoisomerase I inhibitor from natural product
  • Method for fast screening topoisomerase I inhibitor from natural product
  • Method for fast screening topoisomerase I inhibitor from natural product

Examples

Experimental program
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Effect test

Embodiment 2

[0141] Described topoisomerase I inhibitor is natural product extract or its monomeric compound; The present invention preferably natural product lycoris alkaloid extract comprises the following steps:

[0142] ①Preparation of samples to be tested

[0143] Accurately weigh an appropriate amount of the sample to be tested, fully dissolve it with the buffer prepared in Example 1, and the final concentration of the test solution is 2.0 mg / ml;

[0144] The sample to be tested is Lycoris alkaloid extract;

[0145] ②Preparation of standard solution

[0146] Use acetonitrile to prepare the internal standard sample into a standard mother solution with a concentration of 100 μg / ml, add it to the sample solution to be tested before mass spectrometry detection, and the final concentration is 5 μg / ml;

[0147] Described standard substance is nuciferine;

[0148] ③The binding reaction between the test sample and normal and inactive topoisomerase I

[0149] Experimental group: Add 100 m...

Embodiment 3

[0172] The 2015 edition of "Chinese Pharmacopoeia" stipulates that the total amount of diosgenin compound pachyphylloside I, II and pinanoside compound pachyphylloside VI, VII in the medicinal material of papaya should not be less than 0.6% as a measure of its medicinal value. index. In traditional Chinese medicine, Zhonglou is often formulated into prescriptions for the treatment of tumors, such as esophageal cancer, laryngeal cancer, rectal cancer, lung cancer, cervical cancer, leukemia, etc. However, at present, there are few studies on the anti-tumor targets of the Chinese herbal medicine and its main active ingredients.

[0173] The topoisomerase I inhibitor is a natural product extract or a monomeric compound thereof; the topoisomerase I inhibitor described in this embodiment is a monomeric compound saponin I, II, VI and VII;

[0174] ①Preparation of samples to be tested

[0175]Accurately weigh an appropriate amount of the sample to be tested, first dissolve it with a...

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Abstract

The invention discloses a method for fast screening a topoisomerase I inhibitor from a natural product, and relates to the analysis field of natural products. The method comprises the following steps of: (1) preparation of an enzyme binding reaction buffer solution; (2) preparation of a to be detected sample; (3) preparation of a standard solution; (4) binding reaction of the to-be-detected sample and normal and inactive topoisomerase I; (5) chromatography-mass spectrometry detection analysis of reaction sample; and (6) enrichment ratio of the topoisomerase I to the inhibitor. The ultrafiltration membrane technology and the chromatography-mass spectrometry united technology united method are applied to the screening of the topoisomerase I inhibitor, the sample analysis speed is fast and the specificity is strong so as to conveniently recognize a medicine ligand combined with a biological target molecule; meanwhile, the sample analysis dosage is less, and the spectrogram information quantity is large so as to realize the fast screening of the lead compound, the method has great advantage on the aspect of researching mutual effect of medicine small molecule ligand and biological macromolecule receptor; the method is suitable for analyzing in-vitro enrichment ratio of the natural product extract or monomer compound to the topoisomerase I inhibitor.

Description

field of invention [0001] The present invention relates to the field of natural product analysis, in particular to a method for rapidly screening topoisomerase I inhibitors in natural products; specifically, the present invention utilizes ultrafiltration membrane technology combined with chromatography and mass spectrometry to extract A method for rapid screening of topoisomerase I inhibitors in compounds or their monomeric compounds. Background technique [0002] DNA topoisomerase I is an essential enzyme that widely exists in prokaryotic and eukaryotic organisms. The tyrosyl residues in its catalytic site attack DNA single strands, making the enzyme form covalent bonds with DNA 3'-phosphodiester Ligation causes DNA single-strand breaks, thereby affecting DNA topology by regulating supercoiling, linkage, delinkage, and nucleic acid unknotting. Targeted drugs are the most advanced drugs currently used in cancer treatment, which prevent the growth of tumor cells by interacti...

Claims

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Application Information

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IPC IPC(8): G01N30/88
CPCG01N30/88G01N2030/027G01N2030/884
Inventor 郭明全陈桂林田永强张春云
Owner WUHAN BOTANICAL GARDEN CHINESE ACAD OF SCI
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