title: Prediction of in vivo rat biliary drug clearance from an in vitro hepatocyte efflux model.
abstract: Well-established techniques are available to predict in vivo hepatic uptake and metabolism from in vitro data, but predictive models for biliary clearance remain elusive. Several studies have verified the expression and activity of ATP-binding cassette (ABC) efflux transporters central to biliary clearance in freshly isolated rat hepatocytes, raising the possibility of predicting biliary clearance from in vitro efflux measurements. In the present study, short-term plated rat hepatocytes were evaluated as a model to predict biliary clearance from in vitro efflux measurements before major changes in transporter expression known to take place in long-term hepatocyte cultures. The short-term cultures were carefully characterized for their uptake and metabolic properties using a set of model compounds. In vitro efflux was studied using digoxin, fexofenadine, napsagatran, and rosuvastatin, representing compounds with over 100-fold differences in efflux rates in vitro and 60-fold difference in measured in vivo biliary clearance. The predicted biliary clearances from short-term plated rat hepatocytes were within 2-fold of measured in vivo values. As in vitro efflux includes both basolateral and canalicular effluxes, pronounced basolateral efflux may introduce errors in predictions for some compounds. In addition, in vitro rat hepatocyte uptake rates corrected for simultaneous efflux predicted rat in vivo hepatic clearance of the biliary cleared compounds with less than 2-fold error. Short-term plated hepatocytes could thus be used to quantify hepatocyte uptake, metabolism, and efflux of compounds and considerably improve the prediction of hepatic clearance, especially for compounds with a large biliary clearance component.
authors.affiliation: CNS and Pain Innovative Medicines DMPK, AstraZeneca R&D, Södertälje (P.L., J.L., U.F., I.S., J.J., S.B., J.H., L.A.); Cardiovascular and Metabolic Diseases Innovative Medicines DMPK, AstraZeneca R&D, Mölndal (P.L., T.B.A.); Department of Pharmacy, Uppsala University, Uppsala (P.L.); and Section of Pharmacogenetics, Department of Physiology and Pharmacology, Karolinska Institutet, Stockholm (T.B.A.), Sweden.
authors.forename: Patrik
authors.initials: P
authors.lastname: Lundquist
authors.forename: Johan
authors.initials: J
authors.lastname: Lööf
authors.forename: Urban
authors.initials: U
authors.lastname: Fagerholm
authors.forename: Ingemo
authors.initials: I
authors.lastname: Sjögren
authors.forename: Jenny
authors.initials: J
authors.lastname: Johansson
authors.forename: Sveinn
authors.initials: S
authors.lastname: Briem
authors.forename: Janet
authors.initials: J
authors.lastname: Hoogstraate
authors.forename: Lovisa
authors.initials: L
authors.lastname: Afzelius
authors.forename: Tommy B
authors.initials: TB
authors.lastname: Andersson
mesh_terms: D018528:ATP-Binding Cassette Transporters / Q000378:metabolism*; D000818:Animals; D001646:Bile / Q000378:metabolism*; D001798:Blood Proteins / Q000378:metabolism; D002478:Cells, Cultured; D002853:Chromatography, Liquid; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D022781:Hepatocytes / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*; D008297:Male; D008657:Metabolic Clearance Rate; D008862:Microsomes, Liver / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism; D008954:Models, Biological; D004364:Pharmaceutical Preparations / Q000378:metabolism*; D011237:Predictive Value of Tests; D011485:Protein Binding; D051381:Rats; D017207:Rats, Sprague-Dawley; D013379:Substrate Specificity; D053719:Tandem Mass Spectrometry; D013997:Time Factors
chemical_list: D018528:ATP-Binding Cassette Transporters; D001798:Blood Proteins; D004364:Pharmaceutical Preparations; D003577:Cytochrome P-450 Enzyme System