, 2010; van Mil et al., 2004). The close link between BSEP dysfunction and different severe pathophysiological selleck chem inhibitor conditions, in particular the increased susceptibility to DILI, highlights the importance of early identification of clinically relevant drug interactions with BSEP. In this study, we performed a comprehensive analysis of the role that BSEP inhibition plays in DILI and the extent to which it can be used to predict such adverse events prior to large-scale clinical trials. Two in vitro systems of different complexity (inverted membrane vesicles and sandwich-cultured human hepatocytes [SCHH]) were used to determine the influence of drugs on BSEP-mediated taurocholate (TA) transport. Based on our experimental data, we developed a computational model that correctly classified 84% and 91% of the BSEP inhibitors and noninhibitors, respectively.

The clinical impact of BSEP inhibition was evaluated by determining the association between BSEP inhibition and the severity of DILI warnings in drug labels issued by the Food and Drug Administration (FDA). We found BSEP inhibition in membrane vesicles to correlate with DILI severity, and altered disposition of TA in SCHH was shown to distinguish BSEP inhibitors associated with severe DILI from those associated with no or mild DILI. MATERIALS AND METHODS Materials. [3H]-TA was obtained from PerkinElmer (Waltham, MA). Ko143 was a kind gift from Dr Gerrit-Jan Koomen (Van��t Hoff Institute for Molecular Sciences, University of Amsterdam, the Netherlands). GF120918 was kindly provided by GlaxoSmithKline (Stevenage, UK).

MK571 was purchased from A.G. Scientific (San Diego, CA) and astemizole from MP Biomedicals (Eschwege, Germany). Bosentan was purchased from Sequoia Research Products Limited (Pangbourne, UK). All other compounds were purchased from Sigma-Aldrich, St Louis, MO, at the highest purity available (> 95%). Inverted membrane vesicles from Sf9 cells expressing human BSEP (ABCB11) were purchased from SOLVO Biotechnology (Budapest, Hungary). Dulbecco��s modified Eagle��s medium (DMEM) and fetal bovine serum were purchased from Invitrogen, Carlsbad, CA. Dexamethasone, l-glutamine, penicillin, streptomycin, insulin, transferring, and selenium were purchased from Sigma-Aldrich. Hepatocyte maintenance medium (HMM) was obtained from Lonza, Basel, Switzerland and BD Matrigel Basement Membrane Matrix, phenol red free, was purchased from BD Biosciences (Bedford, MA).

Standard Hanks’ balanced salt solution (HBSS) and Ca2+- and Mg2+-free HBSS were purchased Carfilzomib from Invitrogen. Data sets. In the membrane vesicle screen to identify BSEP inhibitors, compounds were selected for inclusion with the intention of optimizing the structural diversity of the data set. The final data set included 250 compounds that were evenly distributed throughout the physicochemical space of registered drugs (Fig. 1). FIG. 1. Chemical diversity of the studied compounds.