Automated Luminescence-Based Cytochrome P450 Profiling Using a Simple, Elegant Robotic Platform

The determination of inhibitory effects that lead compounds have on cytochrome P450 (CYP) ezymes is an important part of today's drug discovery process. Assays can be performed early in the discovery process to predict adverse drug—drug interactions caused by CYP inhibition and to minimize the costs associated with terminating candidates in late stage development or worse, removing a drug from the market after launch. For early discovery work, testing substantial numbers of compounds is desirable, thus automated “mix and read” assays are beneficial. Here, we demonstrate the automation of the CYP profiling process using a simple, yet robust robotic platform. Compound titration, as well as transfer of compounds and assay components was performed by the same automated pipetting system. IC50s of small molecule drugs were determined using recombinant CYP enzymes, CYP3A4, −2C9, and −2D6 and luminogenic substrates specific to each. Compounds were profiled against all three enzymes on the same 384-well assay plate.