posted on 2020-02-28, 21:31authored byGeorge D. Hartman, Scott D. Kuduk, Christine Espiritu, Angela M. Lam
Herein is reported a novel screening
paradigm PURE (P450s under restriction) for the identification and optimization
of hits as part
of a hepatitis B virus (HBV) antiviral discovery program. To closely
represent in vivo hepatocytes, differentiated HepaRG
cells (dHRGs) and primary human hepatocytes (PHHs) were used as the
basis for an HBV infection system. However, a significant challenge
arose during potency evaluation in using cultured dHRGs and PHHs as
screening platforms because, as with hepatocytes in vivo, these cells express active cytochrome P450 enzymes and thus can
metabolize test compounds. The observed antiviral effects may be the
cumulative result of a dynamic pool of parent compound and metabolites
thus confounding structure activity relationship (SAR) interpretation
and subsequent optimization design initiatives. We show here that PURE methodology restricts metabolism of HBV-infected dHRGs
and PHHs and thus provides highly informative potency data for decision-making
on key representative antiviral compounds.