Immobilization Of CYP3A Enzyme By Microencapsulation For In Vitro Drug Interaction Studies

Drug interactions are one of the main causes of adverse drug reactions,and a large number of patients die each year due to serious drug interactions.With advances in pharmacogenomics,the definition of drug-drug interactions is expanding to include drug-gene interactions（DGIs）and drug-drug-gene interactions（DDGIs）.Drug-gene interactions have been incorporated into some treatment guidelines,but research on drug-drug-gene interactions is limited.Cytochrome P450 enzymes play an important role in drug-drug interactions,while little is known about the role of cytochrome P450 enzymes in drug-drug-gene interactions（DDGI）.At present,there are two problems in traditional in vitro drug interaction studies.One is that human liver microsomes,an in vitro drug interaction model containing almost all cytochrome P450 enzymes,are derived from the liver and cannot be regenerated.Second,traditional in vitro drug interaction studies did not take into account the influence of cytochrome P450 gene polymorphisms,which limited the accuracy of the results that were finally extrapolated to the population.Research in（DDGI）is very limited.For this reason,this paper selected the most important CYP3A enzyme among cytochrome P450 enzymes for research,and through meta-analysis,the specific substrate of CYP3A,docetaxel,was used to preliminarily explore the role of CYP3A gene polymorphism in drug-drug-gene interaction（DDGI）.A fixed human liver microsome model that can be used repeatedly and can be used for drug interactions in vitro was constructed by the microencapsulation method.In the future,this model can be used to further study the role of cytochrome P450 gene polymorphisms in drug-drug-gene interactions（DDGI）.Meta-analysis results showed that when docetaxel was used alone,CYP3A gene polymorphism was not associated with its adverse reactions,but when docetaxel was used in combination with other drugs,CYP3A5 A6986G（rs776746）gene polymorphism was associated with hematology.There was a significant correlation with toxicity.Next,the human liver microsomes were immobilized by the microencapsulation method,and the optimal immobilization conditions were determined by single factor and orthogonal experiments.And the remaining activity remained above 60%.Finally,the IC₅₀ values of ketoconazole on CYP3A enzyme before and after immobilization were determined by experiments,indicating that the immobilized CYP3A enzyme can be used for in vitro drug interaction studies.In conclusion,this paper preliminarily determined that the CYP3A5 gene polymorphism in CYP3A enzyme may play a role in drug-drug-gene interaction（DDGI）through meta-analysis,and successfully constructed a reusable and in vitro drug-drug interaction microarray.Capsules immobilize human liver microsome models,which achieves the purpose of saving precious liver sources.This will lay the foundation for further detailed study of the role of cytochrome P450 gene polymorphisms in drug-drug-gene interactions（DDGI）.