Methodological uncertainty in quantitative prediction of human hepatic clearance from in vitro experimental systems

Curr Drug Metab. 2009 Mar;10(3):307-21. doi: 10.2174/138920009787846341.

Abstract

Mechanistic prediction of unbound drug clearance from human hepatic microsomes and hepatocytes correlates with in vivo clearance but is both systematically low (10 - 20 % of in vivo clearance) and highly variable, based on detailed assessments of published studies. Metabolic capacity (Vmax) of commercially available human hepatic microsomes and cryopreserved hepatocytes is log-normally distributed within wide (30 - 150-fold) ranges; Km is also log-normally distributed and effectively independent of Vmax, implying considerable variability in intrinsic clearance. Despite wide overlap, average capacity is 2 - 20-fold (dependent on P450 enzyme) greater in microsomes than hepatocytes, when both are normalised (scaled to whole liver). The in vitro ranges contrast with relatively narrow ranges of clearance among clinical studies. The high in vitro variation probably reflects unresolved phenotypical variability among liver donors and practicalities in processing of human liver into in vitro systems. A significant contribution from the latter is supported by evidence of low reproducibility (several fold) of activity in cryopreserved hepatocytes and microsomes prepared from the same cells, between separate occasions of thawing of cells from the same liver. The large uncertainty which exists in human hepatic in vitro systems appears to dominate the overall uncertainty of in vitro-in vivo extrapolation, including uncertainties within scaling, modelling and drug dependent effects. As such, any notion of quantitative prediction of clearance appears severely challenged.

Publication types

  • Research Support, Non-U.S. Gov't
  • Review

MeSH terms

  • Cytochrome P-450 Enzyme System / chemistry
  • Cytochrome P-450 Enzyme System / physiology
  • Hepatocytes / metabolism
  • Humans
  • Liver / metabolism*
  • Metabolic Clearance Rate
  • Microsomes, Liver / metabolism
  • Models, Biological

Substances

  • Cytochrome P-450 Enzyme System