October 3, 2018

Assay System: 

  • Plated cryopreserved human hepatocytes from 3 individual donors
  • Induction of CYP1A2, CYP2B6 and CYP3A4 by test compound measured by comparing enzyme activity of hepatocytes treated with test compound with vehicle control
  • Optionally, test compound concentration can be monitored during treatment in accordance with 2017 FDA guidance “In Vitro Metabolism- and Transporter-Mediated Drug-Drug Interaction Studies”

Assay Conditions: 

Test Compound Concentration30 µM, 3 µM, and 0.3 µM
Percent Organic Solvent – Inducer Treatment0.1% DMSO
Percent Organic Solvent – Substrate Incubation1.5% acetonitrile
Inducer Treatment Time48 hours
Substrate Incubation Time30 minutes
Treatment and Incubation Conditions37 °C, 5% CO2, and saturating humidity
Positive ControlOmeprazole (1A2), Phenobarbital (2B6), and Rifampicin (3A4)
AnalysisLC-MS/MS
Number of Repeats3

Deliverable: Mean fold induction of test compound and positive control compound.

Test Compound Requirement: 2-3 mg dry powder or 100-200 µL of 30 mM DMSO stock solution.

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