A review of LC-MS techniques and high-throughput approaches used to investigate drug metabolism by cytochrome P450s

Abstract

One of the major challenges facing the pharmaceutical industry today is finding new ways to increase productivity, decrease costs whilst still ultimately developing new therapies that enhance human health. To help address these challenges the utilisation of analytical technologies and high-throughput automated platforms has been employed; in order to perform more experiments in a shorter time frame with increased data quality. One of the main in vitro techniques to assess new chemical entities in a discovery setting has been the use of recombinant liver enzymes, microsomes and hepatocytes. These techniques can help predict in vivo metabolism, clearance and potential drug-drug interactions of these new compounds by cytochrome P450s (the major drug metabolising enzymes). This in vitro methodology has been totally transformed in recent times by the use of automated liquid handling and HPLC tandem mass spectrometry detection techniques (LC-MS/MS). This review aims looking at recent advances in the methodology used to investigate drug metabolism by cytochrome P450s; including an up to date summary of high-throughput platforms including the use of automation and LC-MS/MS to facilitate greater throughput, chromatographic resolution and data quality.