Automated 384-Well Cell-Based Cytochrome P450 Inhibition Assays Using Cryopreserved Human Hepatocytes In Suspension

By Brad Larson and Peter Banks, BioTek Instruments, Inc.
Timothy Moeller, Celsis IVT
Tracy Worzella, Mary Sobol, Dongping Ma, James J. Cali, Promega Corporation

Drug-drug interactions (DDI) are a serious concern to pharmaceutical manufacturers and regulatory agencies. As such, ADME-Tox studies including DDI are tested early in the xenobiotic life cycle. Cytochrome P450 (CYP) enzymes are key drug metabolizers in the body, and therefore play a major role in DDI research and testing. Here we demonstrate an automated assay system, including a luminescent CYP inhibition assay, primary hepatocytes and automated microplate instrumentation, to test lead compound inhibitory effects on CYP isoforms. Validation and pharmacology data, including a comparison to data generated using human liver microsomes, prove that the combination of cells, assay, and instrumentation provide rapid, dependable information on CYP-based drug inhibition in a cell-based format.