Inhibition potential against CYP isoforms (IC50) measured after pre-incubation with and without NADPH in the presence of different concentrations of test compound by monitoring activity of individual CYP enzymes using isoform-specific probe substrates with LC-MS/MS detection.
Time-dependent inhibition (IC50 shift) is determined by comparing IC50 values determined after pre-incubation with and without NADPH.
Test Compound Concentration
Eight concentrations ranging from 0.05 µM to 100 µM
Percent Organic Solvent in Incubation
CYP Isoforms Available for Testing
CYP1A2, CYP2C9, CYP2C19, CYP2D6, and CYP3A4
10-20 minutes (depending on CYP isoform)
Number of Repeats
IC50 of test compound and positive control compound both with and without NADPH pre-incubation.
IC50 shift (IC50 -NADPH/ IC50 +NADPH)
Test Compound Requirement: 1-2 mg dry powder or 100-200 µL of 10 mM acetonitrile stock solution.