Determination of Human Hepatocyte Intrinsic Clearance for Slowly Metabolized Compounds: Comparison of a Primary Hepatocyte/Stromal Cell Co-culture with Plated Primary Hepatocytes and HepaRG

Bonn, Britta; Svanberg, Petter; Janefeldt, Annika; Hultman, Ia; Grime, Ken

doi:10.1124/dmd.115.067769

Cited by 79 publications

(55 citation statements)

References 26 publications

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“…The underprediction for CYP2D6 metabolized compounds in HepaRG™ has already been reported due to a CYP2D6-deficient donor (11,30,31). However, the fold of underprediction seems to vary between the applied enzyme markers due to potential metabolism by multiple CYP enzymes (11,(30)(31)(32). For example, the intrinsic clearances in HepaRG™ as compared to suspension cultures were decreased more for dextromethorphan and metoprolol than for propranolol (11,30).…”

Section: Metabolic Activity Assessment Of In Vitro Liver Models and Cmentioning

confidence: 93%

“…The in vitro clearances were then scaled by applying the well-stirred model using the conventional approach as described in BMATERIALS AND METHODS^(Eqs. [8][9][10][11][12]. Fraction unbound values in the incubation media and human plasma were experimentally determined and are listed in Table III together with the blood-to-plasma ratios.…”

Section: Resultsmentioning

confidence: 99%

“…However, pronounced differences in metabolic activities were observed for CYP2D6. The underprediction for CYP2D6 metabolized compounds in HepaRG™ has already been reported due to a CYP2D6-deficient donor (11,30,31). However, the fold of underprediction seems to vary between the applied enzyme markers due to potential metabolism by multiple CYP enzymes (11,(30)(31)(32).…”

Section: Metabolic Activity Assessment Of In Vitro Liver Models and Cmentioning

confidence: 97%

“…[8][9][10][11][12][13][14][15][16]. For that, the fractions unbound in the incubation media and human plasma were experimentally determined and are listed in Table III together with the blood-to-plasma ratios.…”

Section: Ivive Of In Vitro Clearance Assessmentsmentioning

confidence: 99%

“…Here, they reported the superiority of the HepatoPac® system compared to suspension cultures for predicting low in vivo clearances. Recently, Bonn et al (11) reported the successful application of plated hepatocytes, HepaRG™ and HμREL™ co-cultures, for low intrinsic clearance determination and found slightly better performance of the HμREL™ system. In this study, we report the metabolic activity across a diverse set of phase I and phase II enzyme markers in different in vitro liver models as this is an important prerequisite for successful clearance determination across structural classes.…”

Section: Introductionmentioning

confidence: 99%

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Metabolic Profiling of Human Long-Term Liver Models and Hepatic Clearance Predictions from In Vitro Data Using Nonlinear Mixed-Effects Modeling

Kratochwil

Meille

Fowler

et al. 2017

AAPS J

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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.KEY WORDS: in vitro clearance; in vitro liver models; IVIVE; nonlinear mixed-effects modeling.

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