title: Hepatocyte-based in vitro model for assessment of drug-induced cholestasis.
abstract: Early detection of drug-induced cholestasis remains a challenge during drug development. We have developed and validated a biorelevant sandwich-cultured hepatocytes- (SCH) based model that can identify compounds causing cholestasis by altering bile acid disposition. Human and rat SCH were exposed (24-48h) to known cholestatic and/or hepatotoxic compounds, in the presence or in the absence of a concentrated mixture of bile acids (BAs). Urea assay was used to assess (compromised) hepatocyte functionality at the end of the incubations. The cholestatic potential of the compounds was expressed by calculating a drug-induced cholestasis index (DICI), reflecting the relative residual urea formation by hepatocytes co-incubated with BAs and test compound as compared to hepatocytes treated with test compound alone. Compounds with clinical reports of cholestasis, including cyclosporin A, troglitazone, chlorpromazine, bosentan, ticlopidine, ritonavir, and midecamycin showed enhanced toxicity in the presence of BAs (DICI≤0.8) for at least one of the tested concentrations. In contrast, the in vitro toxicity of compounds causing hepatotoxicity by other mechanisms (including diclofenac, valproic acid, amiodarone and acetaminophen), remained unchanged in the presence of BAs. A safety margin (SM) for drug-induced cholestasis was calculated as the ratio of lowest in vitro concentration for which was DICI≤0.8, to the reported mean peak therapeutic plasma concentration. SM values obtained in human SCH correlated well with reported % incidence of clinical drug-induced cholestasis, while no correlation was observed in rat SCH. This in vitro model enables early identification of drug candidates causing cholestasis by disturbed BA handling.
authors.affiliation: Drug Delivery and Disposition, KU Leuven Department of Pharmaceutical and Pharmacological Sciences, O&N2, Herestraat 49 - bus 921, 3000 Leuven, Belgium. Electronic address: Sagnik.Chatterjee@pharm.kuleuven.be.
authors.forename: Sagnik
authors.initials: S
authors.lastname: Chatterjee
authors.affiliation: KaLy-Cell, 20A rue du Général Leclerc, 67115 Plobsheim, France. Electronic address: l.richert@kaly-cell.com.
authors.forename: Lysiane
authors.initials: L
authors.lastname: Richert
authors.affiliation: Drug Delivery and Disposition, KU Leuven Department of Pharmaceutical and Pharmacological Sciences, O&N2, Herestraat 49 - bus 921, 3000 Leuven, Belgium. Electronic address: Patrick.Augustijns@pharm.kuleuven.be.
authors.forename: Patrick
authors.initials: P
authors.lastname: Augustijns
authors.affiliation: Drug Delivery and Disposition, KU Leuven Department of Pharmaceutical and Pharmacological Sciences, O&N2, Herestraat 49 - bus 921, 3000 Leuven, Belgium. Electronic address: Pieter.Annaert@pharm.kuleuven.be.
authors.forename: Pieter
authors.initials: P
authors.lastname: Annaert
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D002779:Cholestasis / Q000139:chemically induced* / Q000473:pathology; D004305:Dose-Response Relationship, Drug; D022781:Hepatocytes / Q000187:drug effects* / Q000473:pathology; D006801:Humans; D051381:Rats; D015262:Xenobiotics / Q000633:toxicity*
chemical_list: D015262:Xenobiotics
keywords: ALP; ALT; BAs; BSEP/Bsep; Bile acids; Bile salt export pump; CA; DICI; DMSO; Drug-induced cholestasis; FBS; GCDCA; GDCA; HBSS; Hanks' balanced salt solution; NOAEL; NTCP/Ntcp; PBS; SCH; SCHH; SCRH; SM; Safety margin; Sandwich-cultured hepatocytes; TCA; ULN; Urea assay; alanine aminotransferase; aspartate aminotransferase; bile acids; bile salt export pump (human/rat); cholic acid; dimethyl sulfoxide; drug-induced cholestasis index; fetal bovine serum; glycochenodeoxycholic acid; glycodeoxycholic acid; no observed adverse effect level; phosphate buffered saline; safety margin; sandwich-cultured hepatocytes; sandwich-cultured human hepatocytes; sandwich-cultured rat hepatocytes; sodium taurocholate cotransporting polypeptide (human/rat); taurocholic acid; upper limit of normal