High-throughout medicament screening method built on primary hepatocytes serving as carrier

Abstract

The invention discloses a high-throughout medicament screening method built on primary hepatocytes serving as a carrier, which relates to the field of biotechnology. After a medicament and a substrate are incubated together, liquid-chromatogram connection is adopted to detect two or more than two kinds of metabolins at the same time. The method comprises: (a), culturing primary hepatocytes by a 'sandwich gel' process; (b) adding a medicament to be screened and a corresponding positive inducer, culturing for three days, and adding at least two or more than two kinds of substrates for metabolism evaluation; and (c) establishing a liquid phase and tandem mass spectrum combined process capable of detecting two or more than two kinds of metabolins at the same time, evaluating the influence of the medicament on the activity of hepatocytes by detecting the two or more than two kinds of metabolins at the same time by liquid-chromatogram connection, and thus evaluating if there is a potential medicament application risk from an overall perspective.

Description

Technical field: [0001] The invention relates to the field of biotechnology, in particular to a high-throughput drug screening method based on liver primary cells as carriers. Background technique: [0002] In addition to the liver removing the drug in the body through metabolic decomposition, the drug can also be taken up by the liver cells and then excreted into the bile, and then excreted by excreting the compound into the feces along with the excretion of the bile. These activities of drug uptake and bile elimination are usually carried out by the carrier transport system in hepatocytes. The drugs and other compounds excreted by the carrier in the liver cells first enter the hepatic sinusoids in the space of the hepatic cells, pass through the hepatic sinusoids, enter the bile ducts, and then are excreted with the bile. Therefore, the degree of drug excretion through bile is also of great significance to the research of drugs. [0003] Due to the emergence and rapid de...

AI Technical Summary

Problems solved by technology

The simple detection of the effect on the activity of a single enzyme is far from meeting the requirements, and it is more and more urgent to establish a method that can simultaneously evaluate the effect of the test compound on at least two or more cytochrome CYP450 enzymes.

[0005] The existing technology at home and abroad using liver primary cells for preclinical drug screening research only uses the biological characteristics of liver cells, and detects the activity of single cytochrome CYP450 enzymes in liver cells, without the main cytochrome CYP450 Enzyme Activity Evaluation Study

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