Method for screening active component with function of hexokinase inhibition effects

A technology of hexokinase and inhibition, applied in the field of screening of active ingredients of traditional Chinese medicines, can solve the problems of tumor cell apoptosis, inability to obtain glucose 6-phosphate, etc., and achieve the effects of less dosage, saving of biochemical reagents, and rapid screening

Inactive Publication Date: 2012-02-01
FUDAN UNIV
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Problems solved by technology

In view of the significance of hexokinase to the activities of various tumor cells, the researchers reasonably speculate that if an effective hexokinase inhibitor can be found and the activity of the enzyme can be inhibited, the glycolysis of tumor cells will be inhibited, and tumor cells will be inhibited. Without enough glucose 6-phosphate, ATP cannot be converted into ADP to provide energy for various activities of cells, which in turn leads to apoptosis of tumor cells

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  • Method for screening active component with function of hexokinase inhibition effects
  • Method for screening active component with function of hexokinase inhibition effects
  • Method for screening active component with function of hexokinase inhibition effects

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Embodiment 1

[0027] Example 1: Screening of Hexokinase Inhibitory Active Components in the Extract of Squilla Sinensis

[0028] 1. Determination of ATP and ADP by LC-MS

[0029] Chromatographic column: Agilent Extend C-18 (3.1×100mm, 3μm; Agilent, Palo Alto, CA, USA); mobile phase: 5mM ammonium acetate (pH7.5):methanol=6:4; flow rate: 0.3mL / min; Column temperature: 20°C; Injection volume: 10μL

[0030] Ion source: liquid chromatography-mass spectrometry interface: electrospray interface (ESI), positive ion mode (positive ion); drying gas flow rate: 10mL / min; drying gas temperature: 300°C; atomizing gas pressure: 30psi (upper limit 60psi); capillary voltage: 3000V; mass analyzer: detection method: selected ion monitoring (SIM); detection interval: 0-1.6min; fragment voltage: 100V; monitoring ion: ATP, quasi-molecular ion peak (m / z 508) ;ADP, quasi-molecular ion peak (m / z428)

[0031] Specific as figure 1 As shown, the ATP retention time is 1.39 minutes, and the ADP retention time is 1.4...

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Abstract

The invention belongs to the field of drug analysis, relates to a method for screening an active component in a traditional Chinese medicine, and specifically, relates to a method for screening a traditional Chinese medicine active component with a function of hexokinase inhibition effects. Based on hexokinase catalytic reaction adopting adenosine triphosphate (ATP) as a substrate, the method can screen out a traditional Chinese medicine active component with a function of hexokinase inhibition effects through quantitative analysis processes on ATP and product ATP. The method is rapid, simple, economic and effective. Compared with a cell test method, the method reduces an extract use amount, saves biochemical reagents, and shortens a screening period. Therefore, the method has great importance for screening and research of traditional Chinese medicine active components. In the invention, all experiment parameters of an improved liquid chromatography-mass spectrometry (LC-MS) method have good reference values for mass spectrometry technology-based determination of other biomolecules with similar properties.

Description

technical field [0001] The invention belongs to the field of drug analysis, and relates to a screening method for active components in traditional Chinese medicinal materials, in particular to a screening method for active components of traditional Chinese medicines with hexokinase inhibitory effect. Background technique [0002] Malignant tumor is a difficult disease that seriously threatens human life, and it is the second killer after heart disease. At present, the commonly used clinical drug treatment is chemotherapy, however, chemotherapy often brings serious side effects. Therefore, finding and screening anti-tumor active ingredients from natural resources and developing effective and low-toxic drugs have attracted widespread attention of medical workers. However, practice has proved that the components of natural medicinal materials, such as Chinese medicinal materials, are very complex, which brings great challenges to the screening of active ingredients. It is nec...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K36/42G01N30/02G01N30/72A61P35/00
Inventor 郁韵秋潘乐
Owner FUDAN UNIV
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