Nitrosamine TD50 prediction method based on fluorescent yeast sensor

Abstract

The invention discloses a nitrosamine TD50 prediction method based on a fluorescent yeast sensor. The method comprises the following steps: monitoring the expression level of the GFP fusion protein exposed to a nitrosamine compound in real time by using a DNA damage response fluorescent yeast sensor capable of expressing the GFP fusion protein, and quantifying the genetic toxicity by adopting a TEL (Total Expression Level); then according to the TEL 1.5 of the nitrosamine compound, predicting the TD50 of the nitrosamine compound without TD50 data report. The method has the advantages of being low in cost, time-saving, reliable in information and small in sample dosage, can be widely applied, and is beneficial to improving the research and development speed and quality of drugs.

Description

technical field [0001] The invention belongs to the field of drug analysis and detection, and relates to a nitrosamine TD based on a fluorescent yeast sensor 50 method of prediction. Background technique [0002] During the production and storage of pharmaceuticals, potentially mutagenic and carcinogenic impurities, namely genotoxic impurities (GTIs), may be introduced. Genotoxic impurities often cause induction of DNA adducts, strand breaks, point mutations, and structural and numerical changes in chromosomes by interacting with DNA or other targets that control apoptosis [1-3] . Genotoxic impurities can induce gene mutation and genetic material damage at low concentrations, and further lead to the occurrence of cancer [4] . [0003] Drug recalls related to nitrosamine impurities have aroused the attention of the global pharmaceutical industry and global drug regulatory agencies, and the regulation of genotoxic impurities represented by nitrosamine impurities has become...

AI Technical Summary

Problems solved by technology

This method evaluates genotoxicity by observing the migration level of damaged DNA in electrophoresis after the action of the compound, and has the disadvantage of limited detection of damage types

Computer simulation evaluation based on compound warning structure is a commonly used method for genotoxicity evaluation of drug impurities in actual production, but this method is prone to false positive or false negative results, resulting in a sharp increase in research and development costs and hidden dangers to drug quality and safety.

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