Qualitative and quantitative prediction of human in vivo metabolic pathways in a human hepatocyte-murine stromal cell co-culture model

Abstract

1. This study assessed the value of a static in vitro human hepatocyte-murine stromal cell co-culture model to qualitatively and quantitatively predict human in vivo metabolic clearance pathways using ¹⁴C-labeled test compounds and compared these results to an in vitro suspended human hepatocyte model and the in vivo human ¹⁴C ADME studies.

2. Test compounds represented a diverse set of clearance pathways (Phase I and Phase II). Compounds were incubated for 4 h in suspended human hepatocytes and for 24 and 168 h in the human co-culture model. Multivariate analysis revealed that long-term (168 h) incubation of test compounds in the co-culture had reasonable quantitative prediction of the in vivo human clearance pathways as compared to the 4 h suspended hepatocytes or the 24 h co-culture incubation.

3. In vivo and in vitro disconnects were observed in cases where extra-hepatic metabolism or urinary excretion was observed in vivo. Differences in the relative percentages of Phase I and Phase II metabolites observed were likely due to microbial β-glucuronidase hydrolysis of conjugates and microflora mediated metabolism in the gut not present in the in vitro systems.

Keywords:

• ¹⁴C human
• ¹⁴C-labeled compounds
• hepatocytes
• hepatocyte-stromal cell co-culture
• metabolite prediction
• metabolites

Declaration of interest

This work was supported by Eli Lilly and Company. Cassidy and Yi are employees and shareholders of Eli Lilly and Co.