CYP450 inhibition studies are an essential part of drug discovery to predict in vivo drug-drug interactions.
Inhibition of cytochrome P450 enzyme catalytic activity is a leading mechanism of metabolism-based drug-drug interactions. Many DDI-related pre-clinical/clinical failures to date have been attributed to CYP450-related hepatic metabolism. Hence it becomes essential to screen compounds in early drug discovery to monitor their in vitro inhibition of the major CYP450 isoforms in order to predict their in vivo effects.
Despite the many CYP450 isozymes present in man, five major isoforms (3A4, 2C9, 2C19, 2D6, 1A2) account for the metabolism-related DDI issues for greater than 90% of marketed drugs.
Commonly such assays probe the metabolism-related DDI of drug candidates by monitoring the effect of the test compounds on CYP450 metabolic activity using a known substrate. The FDA and EMA recommend many specific drug substrates for CYP450 isoforms.
CYP450 Inhibition: Standard Assay Conditions (Customizable)
|CYP Isoforms||CYP1A2, CYP2B6, CYP2C8, CYP2C9, CYP2C19, CYP2D6 and CYP3A4 (other isoforms are available)|
|Matrix||Pooled Whole Human Liver Microsomes|
CYP1A2-Tacrine (only for RapidFire-MS/MS)
|Testing Compound Concentration||0, 0.2, 0.5, 1.3, 3.2, 8, 20, 50 μM|
|Detection Method||Specific metabolites by LC-MS/MS or RapidFire-MS/MS|
|Deliverables||IC50, Standard error of IC50, Report|
|Turnaround Time||2-3 Days|
|Compound requirements||100 μL 50 mM DMSO stock or 1-2 mg solid|