Human cytochrome P450 (CYP450) enzymes play an essential role in drug metabolism. Even subtle changes in the activities of CYP enzymes may cause severe drug-drug interactions.

Cytochrome P450 (CYP) is a superfamily of heme-containing enzymes, which mediate the inactivation and metabolism of many drugs and endogenous substances. Compounds that inhibit P450 may cause the accumulation of toxicity of other substrates. CYP Cytochrome P450 inhibition assay examines the effects of test compounds on the metabolism of five major human metabolic CYP substrates: 1A2, 2B6, 2C9, 2D6, and 3A4. The levels of labeled substrates and metabolites of each CYP isoform are measured by LC/MS/MS in the presence and absence of test compounds.

Cytochrome P450 Test (CYP) Inhibition Assay Protocol (Single Point)

Compound test concentration10µM,0µM
CYP450 isoforms1A2,3A4, 2C9, 2C19, 2D6
Microsome concentration0.05-0.5mg/mL
Incubation duration5-40min
IC50 calculation
Data InterpretationIC50<1 µM Strong Inhibitor
1µM IC50>10 µM Weak or Non Inhibitor

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