CYP inhibition, hepatocytes

The test compound is preincubated with human hepatocytes for 0 and 30min. Thereafter, CYP activity is measured by incubating the hepatocytes with a cocktail of CYP-specific substrates (1A2, 2B6, 2C8, 2C9, 2D6, 2C19 and 3A). Direct inhibition is observed from the samples preincubated for 0 min as reduced level of CYP-specific metabolite formation in the incubations compared to solvent samples (IC50 determination). Time-dependent (TDI) inhibition is detected as changes in inhibition curves after preincubation for 0 and 30 min (IC50 shift determination).
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Test concentrations: 6

Time points: 1

Replicates: 2

Positive controls

UPLC/MS/MS analysis

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Direct inhibition as % inhibition or as IC50 for each CYP, tentative Ki values, time-dependent inhibition as IC50 shift or as % inhibition

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Full Word/PDF report

Background

Inhibition of cytochrome P450 (CYP) enzymes by a new chemical entity (NCE) may decrease the metabolism of co-medicated drugs. We offer assays for individual enzymes as well as a cocktail approach to determine inhibition towards multiple enzymes in the same incubation. Liver microsomes are recommended for high-throughput screenings and for mechanistic approach. In addition, CYP inhibition evaluation can be conducted using pooled hepatocytes, for compounds where hepatocytes are a better suited model.

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