High-Throughput Evaluation of CYP1A1 and 2B1 Induction in Rat Liver Slices Using a Semi-Automated System

Jairam   R.   Palamanda; Xinjie      Lin; Pramila      Kumari; Amin   A.   Nomeir

Abstract

Drug candidates with the propensity to induce rat CYP1A1 or 2B1 isoforms are believed to possess a greater tendency to induce hepatic tumors in oncogenicity studies. We have previously published on a manual rat liver slice assay that showed a satisfactory relationship between in vitro CYP2B1 m-RNA induction using real time PCR and the ex vivo pentoxyresorufin O-dealkylase (PROD) activity in liver microsomes prepared from rats treated daily via the oral route for 14 consecutive days with inducers or non-inducers. We now describe this automated in vitro high throughput liver slice technique to screen out drug candidates that are potent rodent CYP1A1 and/or CYP2B1 inducers. A good concordance between in vitro and in vivo data was observed for both CYP1A1 (100 %) and CYP2B1 (90%) isoforms. Automation of key steps has enabled us to increase the annual screening throughput from 200 (manual) to 1500 compounds. The increase in throughput allowed the quick development of structure-induction relationships (SIRs) for multiple drug discovery programs in a facile manner.

Keywords: Automation, HTP or High Throughput, Liver Slices, Real Time PCR, Cytochrome P450, Induction, CYP1A1, CYP2B1