title: Metabolic Profiling of Human Long-Term Liver Models and Hepatic Clearance Predictions from In Vitro Data Using Nonlinear Mixed-Effects Modeling.
abstract: Early prediction of human clearance is often challenging, in particular for the growing number of low-clearance compounds. Long-term in vitro models have been developed which enable sophisticated hepatic drug disposition studies and improved clearance predictions. Here, the cell line HepG2, iPSC-derived hepatocytes (iCell®), the hepatic stem cell line HepaRG™, and human hepatocyte co-cultures (HμREL™ and HepatoPac®) were compared to primary hepatocyte suspension cultures with respect to their key metabolic activities. Similar metabolic activities were found for the long-term models HepaRG™, HμREL™, and HepatoPac® and the short-term suspension cultures when averaged across all 11 enzyme markers, although differences were seen in the activities of CYP2D6 and non-CYP enzymes. For iCell® and HepG2, the metabolic activity was more than tenfold lower. The micropatterned HepatoPac® model was further evaluated with respect to clearance prediction. To assess the in vitro parameters, pharmacokinetic modeling was applied. The determination of intrinsic clearance by nonlinear mixed-effects modeling in a long-term model significantly increased the confidence in the parameter estimation and extended the sensitive range towards 3% of liver blood flow, i.e., >10-fold lower as compared to suspension cultures. For in vitro to in vivo extrapolation, the well-stirred model was used. The micropatterned model gave rise to clearance prediction in man within a twofold error for the majority of low-clearance compounds. Further research is needed to understand whether transporter activity and drug metabolism by non-CYP enzymes, such as UGTs, SULTs, AO, and FMO, is comparable to the in vivo situation in these long-term culture models.
authors.affiliation: Roche Pharmaceutical Research and Early Development, Roche Innovation Center Basel, Grenzacherstrasse 124, 4070, Basel, Switzerland. nicole_a.kratochwil@roche.com.
authors.forename: Nicole A
authors.initials: NA
authors.lastname: Kratochwil
authors.affiliation: Roche Pharmaceutical Research and Early Development, Roche Innovation Center Basel, Grenzacherstrasse 124, 4070, Basel, Switzerland.
authors.forename: Christophe
authors.initials: C
authors.lastname: Meille
authors.affiliation: Roche Pharmaceutical Research and Early Development, Roche Innovation Center Basel, Grenzacherstrasse 124, 4070, Basel, Switzerland.
authors.forename: Stephen
authors.initials: S
authors.lastname: Fowler
authors.affiliation: Roche Pharmaceutical Research and Early Development, Roche Innovation Center Basel, Grenzacherstrasse 124, 4070, Basel, Switzerland.
authors.forename: Florian
authors.initials: F
authors.lastname: Klammers
authors.affiliation: Roche Pharmaceutical Research and Early Development, Roche Innovation Center Basel, Grenzacherstrasse 124, 4070, Basel, Switzerland.
authors.forename: Aynur
authors.initials: A
authors.lastname: Ekiciler
authors.affiliation: Roche Pharmaceutical Research and Early Development, Roche Innovation Center Basel, Grenzacherstrasse 124, 4070, Basel, Switzerland.
authors.forename: Birgit
authors.initials: B
authors.lastname: Molitor
authors.affiliation: Roche Pharmaceutical Research and Early Development, Roche Innovation Center Basel, Grenzacherstrasse 124, 4070, Basel, Switzerland.
authors.forename: Sandrine
authors.initials: S
authors.lastname: Simon
authors.affiliation: Roche Pharmaceutical Research and Early Development, Roche Innovation Center Basel, Grenzacherstrasse 124, 4070, Basel, Switzerland.
authors.forename: Isabelle
authors.initials: I
authors.lastname: Walter
authors.affiliation: Roche Pharmaceutical Research and Early Development, Roche Innovation Center Basel, Grenzacherstrasse 124, 4070, Basel, Switzerland.
authors.forename: Claudia
authors.initials: C
authors.lastname: McGinnis
authors.affiliation: Roche Pharmaceutical Research and Early Development, Roche Innovation Center Basel, Grenzacherstrasse 124, 4070, Basel, Switzerland.
authors.forename: Johanna
authors.initials: J
authors.lastname: Walther
authors.affiliation: Roche Pharmaceutical Research and Early Development, Roche Innovation Center Basel, Grenzacherstrasse 124, 4070, Basel, Switzerland.
authors.forename: Brian
authors.initials: B
authors.lastname: Leonard
authors.affiliation: Roche Pharmaceutical Research and Early Development, Roche Innovation Center Basel, Grenzacherstrasse 124, 4070, Basel, Switzerland.
authors.forename: Miriam
authors.initials: M
authors.lastname: Triyatni
authors.affiliation: Roche Pharmaceutical Research and Early Development, Roche Innovation Center Basel, Grenzacherstrasse 124, 4070, Basel, Switzerland.
authors.forename: Hassan
authors.initials: H
authors.lastname: Javanbakht
authors.affiliation: Roche Pharmaceutical Research and Early Development, Roche Innovation Center Basel, Grenzacherstrasse 124, 4070, Basel, Switzerland.
authors.forename: Christoph
authors.initials: C
authors.lastname: Funk
authors.affiliation: Roche Pharmaceutical Research and Early Development, Roche Innovation Center Basel, Grenzacherstrasse 124, 4070, Basel, Switzerland.
authors.forename: Franz
authors.initials: F
authors.lastname: Schuler
authors.affiliation: Roche Pharmaceutical Research and Early Development, Roche Innovation Center Basel, Grenzacherstrasse 124, 4070, Basel, Switzerland.
authors.forename: Thierry
authors.initials: T
authors.lastname: Lavé
authors.affiliation: Roche Pharmaceutical Research and Early Development, Roche Innovation Center Basel, Grenzacherstrasse 124, 4070, Basel, Switzerland.
authors.forename: Neil J
authors.initials: NJ
authors.lastname: Parrott
mesh_terms: D018920:Coculture Techniques; D019389:Cytochrome P-450 CYP2D6 / Q000378:metabolism; D004798:Enzymes / Q000378:metabolism; D056945:Hep G2 Cells; D022781:Hepatocytes / Q000201:enzymology / Q000378:metabolism*; D006801:Humans; D008099:Liver / Q000201:enzymology / Q000378:metabolism*; D008954:Models, Biological; D017711:Nonlinear Dynamics; D004364:Pharmaceutical Preparations / Q000378:metabolism; D010599:Pharmacokinetics; D013997:Time Factors
chemical_list: D004798:Enzymes; D004364:Pharmaceutical Preparations; D019389:Cytochrome P-450 CYP2D6
keywords: IVIVE; in vitro clearance; in vitro liver models; nonlinear mixed-effects modeling