Application of a Substrate Cocktail Approach in the Assessment of Cytochrome P450 Induction Using Cultured Human Hepatocytes

作者：Pelletier Robert D^*; Lai W George; Wong Y Nancy

来源：Journal of Biomolecular Screening, 2013, 18(2): 199-210.

DOI：10.1177/1087057112463732

• CYP induction
• high-throughput screening
• drug-drug interaction

Induction of the cytochrome P450 (CYP) family of enzymes by coadministered compounds can result in drug-drug interactions, as in the case of the coadministration of rifampicin with many CYP3A substrates, including midazolam. Identification of potential drug-drug interactions due to CYP induction during drug discovery is critical. We present a substrate cocktail method that was applied to assess the induction of CYP1A, CYP2B6, CYP2C9, and CYP3A using a 96-well high-throughput format. Viable cell counts were determined using a high-content screening system to normalize activities. Substrate cocktail incubations demonstrated a similar fold induction for known inducers as compared with discrete probe incubations. The system was further validated by determining the induction potency of rifampicin. The E-max and EC50 values in two separate lots of hepatocytes for CYP3A induction by rifampicin in a 96-well format were similar when discrete probe was compared with the probe cocktail. This system has been demonstrated to be suitable for high-throughput assessments of CYP induction.

• 出版日期2013-2