Prediction of fraction metabolized via CYP3A in humans utilizing cryopreserved human hepatocytes from a set of 12 single donors

Abstract

1.  It has previously been demonstrated that metabolism of drugs via a single enzymatic pathway, particularly CYP3A4, is associated with increased risk for drug-drug interactions (DDI). Quantitative experimental systems as well as integrated prediction models to assess such risk during the preclinical phase are highly warranted.

2.  The present study was designed to systematically investigate the performance of human cryopreserved hepatocytes in suspension to predict fraction metabolized via CYP3A (fm_CYP3A) by assessing the ketoconazole sensitive intrinsic clearance (CL_int) for five prototypical CYP3A substrates with varying degree of CYP3A dependent CL_int in twelve individual hepatocyte batches.

3.  We demonstrate that in contrast to well predicted mean hepatic metabolic clearance (CL_H) and mean fm_CYP3A data, the variability in CYP3A contribution for compounds having multiple metabolic pathways cannot be predicted from inhibition experiments using ketoconazole as inhibitor. Instead, data in the present paper indicate that the variability is larger after inhibition of CYP3A for compounds having multiple metabolic pathways.

4.  It is therefore recommended to estimate the average CL_int and fm_CYP3A for a given test compound in a series (n = 10) of individual human hepatocyte batches.

• CYP450 inhibition
• DDI risk
• hepatocytes suspension
• ketoconazole
• preclinical