Bioluminescent Approaches for In Vitro ADMET Scientific Poster
Michael Curtin, James J. Cali, Mary Sobol, Dongping Ma, John Shultz, Michael Valley
Promega Corporation, 2800 Woods Hollow Rd., Madison, WI 53711
ADMET targets were coupled to firefly luciferase in assays that used the convenience of light output as a readout. In a cell-based approach test compounds were tested for glutathione depletion, P450 transcriptional induction and induction of P450 enzyme activity. Inductions of P450 enzyme activities with selective luminogenic substrates were measured and observed as markers for identifying ligands for PXR, aryl hydrocarbon or PPAR-alpha nuclear receptors. Cell-free membrane assays with luminogenic substrates were used to measure P450 or monoamine oxidase enzyme activities and their inhibition by test compounds. IC50s from these bioluminescent assays correlated well with conventional methods in terms of rank order and absolute potency. Each bioluminescent assay relied on the light generating reaction of firefly luciferase, coupling light output with target activity. The assays were insensitive to interference from fluorescent analytes and had low intrinsic background signals giving them high levels of sensitivity and large dynamic ranges for robust, high throughput applications.
- Part# PS066
- Printed in USA.