Application of a Micropatterned Cocultured Hepatocyte System To Predict Preclinical and Human-Specific Drug Metabolism

Abstract

Laboratory animal models are the industry standard for preclinical risk assessment of drug candidates. Thus, it is important that these species possess profiles of drug metabolites that are similar to those anticipated in human, since metabolites also could be responsible for biologic activities or unanticipated toxicity. Under most circumstances, preclinical species reflect human in vivo metabolites well; however, there have been several notable exceptions, and understanding and predicting these exceptions with an in vitro system would be very useful. Human micropatterned cocultured (MPCC) hepatocytes have been shown to recapitulate human in vivo qualitative metabolic profiles, but the same demonstration has not been performed yet for laboratory animal species. In this study, we investigated several compounds that are known to produce human-unique metabolites through CYP2C9, UGT1A4, aldehyde oxidase (AO), or N-acetyltransferase that were poorly covered or not detected at all in the selected preclinical species. To perform our investigation we used 24-well MPCC hepatocyte plates having three individual human donors and a single donor each of monkey, dog, and rat to study drug metabolism at four time points per species. Through the use of the multispecies MPCC hepatocyte system, the metabolite profiles of the selected compounds in human donors effectively captured the qualitative in vivo metabolite profile with respect to the human metabolite of interest. Human-unique metabolites that were not detected in vivo in certain preclinical species (normally dog and rat) were also not generated in the corresponding species in vitro, confirming that the MPCC hepatocytes can provide an assessment of preclinical species metabolism. From these results, we conclude that multispecies MPCC hepatocyte plates could be used as an effective in vitro tool for preclinical understanding of species metabolism relative to humans and aid in the choice of appropriate preclinical models.

Liver safety evaluation of endothelin receptor antagonists using HepatoPac®: A single model impact assessment on hepatocellular health, function and bile acid disposition

“…Xenobiotic metabolic competency was demonstrated in this study and in other studies using different compounds across rat and human MPCCs (Ballard et al, ). Metabolic competency was demonstrated here as products of oxidation reactions coupled with glucuronidation or glucuronidation of the parent alone.…”

Modeling Therapeutic Antibody–Small Molecule Drug-Drug Interactions Using a Three-Dimensional Perfusable Human Liver Coculture Platform

“…These methods have shown promise in the evaluation of hepatic drug metabolism, DILI, and PK (Chao et al, 2009;Kostadinova et al, 2013;Messner et al, 2013;Ballard et al, 2016). The LiverChip platform offers continuous perfusion of oxygenated medium through a scaffold containing microchannels for 3D cell culture under flow.…”

“…Recently, there have been several platforms introduced that offer the potential for long-term, multiweek primary human hepatocyte culture, which would provide researchers the ability to model more complex interactions involving chronic dosing effects, low-clearance compound and drug accumulation, human-specific metabolite identification, DILI, and the effects of chronic inflammation (Chao et al, 2009;Kostadinova et al, 2013;Messner et al, 2013;Ballard et al, 2016). There are several approaches that can increase in vitro hepatic performance and concomitant physiologic relevance, including three-dimensional (3D) cell culture, application of media flow and shear conditions to mimic natural tissue, and cocultures with liver nonparenchymal cell types such as Kupffer cells (KCs), liver sinusoidal endothelial cells, and hepatic stellate cells (LeCluyse et al, 2012;Ebrahimkhani et al, 2014).…”

Metabolites of alectinib in human: their identification and pharmacological activity

“…In general, hepatocytes and liver subcellular fractions are commonly known and predictive approaches to elucidate these steps have been published [8, 9, 10, 11], however, these in vitro approaches have proved insufficient to reflect the in vivo profile and the main metabolic pathway in human in the case of such as slow metabolite turnover rate, and extrahepatic metabolism. Therefore, a human ADME study is essential to determine via mass balance the elimination pathways of an administered drug and give a metabolite profile definitively in human.…”