title: Assessment of hepatotoxicity potential of drug candidate molecules including kinase inhibitors by hepatocyte imaging assay technology and bile flux imaging assay technology.
abstract: Kinases are members of a major protein family targeted for drug discovery and development. Given the ubiquitous nature of many kinases as well as the broad range of pathways controlled by these enzymes, early safety assessments of small molecule inhibitors of kinases are crucial in identifying new molecules with sufficient therapeutic window for clinical development. Failure or attrition of drug candidates in late-stage pipelines due to hepatotoxicity is a significant challenge in the drug development field. Herein we provide detailed methods for the hepatocyte imaging assay technology (HIAT) and the bile flux imaging assay technology (BIAT) to evaluate drug-induced liver injury (DILI) potentials for drug candidates. Optimized culturing methods for primary human hepatocytes, both freshly isolated and prequalified cryopreserved cells, are also presented. The applications of these high-content cellular imaging technologies in the evaluation of p38 and Her2 kinase inhibitors are highlighted to illustrate the usefulness of the research methodology in a compound screening as well as mechanistic investigative setting.
authors.affiliation: Knowledge Discovery & Knowledge Management, Merck & Co., Inc., RY86-235, Rahway, NJ, USA. jinghai_xu@merck.com
authors.forename: Jinghai J
authors.initials: JJ
authors.lastname: Xu
authors.forename: Margaret C
authors.initials: MC
authors.lastname: Dunn
authors.forename: Arthur R
authors.initials: AR
authors.lastname: Smith
authors.forename: Eric S
authors.initials: ES
authors.lastname: Tien
mesh_terms: D001646:Bile / Q000378:metabolism*; D001681:Biological Assay / Q000379:methods; D002478:Cells, Cultured; D003094:Collagen; D015925:Cryopreservation; D004338:Drug Combinations; D004791:Enzyme Inhibitors / Q000633:toxicity*; D022781:Hepatocytes / Q000187:drug effects* / Q000201:enzymology*; D006801:Humans; D007091:Image Processing, Computer-Assisted; D007797:Laminin; D008856:Microscopy, Fluorescence; D057054:Molecular Imaging / Q000379:methods*; D010770:Phosphotransferases / Q000037:antagonists & inhibitors*; D011509:Proteoglycans; D013194:Staining and Labeling; D018675:Toxicity Tests / Q000379:methods*
chemical_list: D004338:Drug Combinations; D004791:Enzyme Inhibitors; D007797:Laminin; D011509:Proteoglycans; C059485:matrigel; D003094:Collagen; D010770:Phosphotransferases