Drug-induced cholestasis assay in primary hepatocytes

Pieter Van Brantegem; Sagnik Chatterjee; Tom De Bruyn; Pieter Annaert; Neel Deferm

doi:10.1016/j.mex.2020.101080

Drug-induced cholestasis assay in primary hepatocytes

MethodsX. 2020 Sep 25:7:101080. doi: 10.1016/j.mex.2020.101080. eCollection 2020.

Authors

Pieter Van Brantegem¹, Sagnik Chatterjee², Tom De Bruyn³, Pieter Annaert¹, Neel Deferm¹

Affiliations

¹ KU Leuven Department of Pharmaceutical and Pharmacological Sciences, Leuven, Belgium.
² Pharmaceutical Candidate Optimization, Biocon, Bristol-Myers Squibb R& D Center (BBRC), Syngene International Ltd., Bangalore, India.
³ Department of Drug Metabolism and Pharmacokinetics, Genentech Inc, South San Francisco, CA, USA.

Abstract

Drug-induced cholestasis (DIC) is a major cause of clinical failure of drug candidates. Numerous patients worldwide are affected when exposed to marketed drugs exhibiting a DIC signature. Prospective identification of DIC during early compound development remains challenging. Here we describe the optimized in vitro procedure for early assessment and prediction of an increased DIC risk. Our method is based on three principles:•Exposure of primary human hepatocyte cultures to test compounds in the absence and presence of a physiologically relevant mixture of endogenous bile salts.•Rapid and quantitative assessment of the influence of concomitant bile salt exposure on hepatocyte functionality and integrity after 24 h or 48 h of incubation.•Translation of the in vitro result, expressed as a DIC index (DICI) value, into an in vivo safety margin.Using our historical control data, a new (data driven) DICI cut-off value of 0.78 was established for discerning cholestatic and non-cholestatic compounds. Our DIC assay protocol was further improved by now relying on the principle of the no observable adverse effect level (NOAEL) for determining the highest test compound concentration corresponding to a DICI ≥ 0.78. Predicted safety margin values were subsequently calculated for compounds displaying hepatotoxic and/or cholestatic effects in patients, thus enabling evaluation of the performance of our DIC assay. Of note, this assay can be extended to explore the role of drug metabolites in precipitating DIC.

Keywords: Drug candidate selection; Drug safety evaluation; Drug-induced cholestasis; Drug-induced liver injury; In vitro toxicity; Primary hepatocytes.