Addressing the Challenges of Low Clearance in Drug Research

Di, Li; Obach, R. Scott

doi:10.1208/s12248-014-9691-7

Cited by 76 publications

(75 citation statements)

References 20 publications

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“…In this range, the clearance values can be determined with confidence, e.g., with a coefficient of variation (CV) of ≤30% for multiple determinations. As the maximal incubation time of the hepatocyte suspension cultures is 2-4 h as compared to microsomes (0.5-1 h), the sensitive range for in vitro clearance determination in hepatocyte suspension cultures is lower, e.g., between 3 and 200 μL min −1 mg −1 protein (CV ≤ 30%) (4). For the majority of drugs, the volume of distribution is between the blood volume of 0.1 and 10 L/kg (28).…”

Section: Discussionmentioning

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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Section: Discussionmentioning

confidence: 99%

“…protein depending on the physicochemical properties of the compounds (BPP and fu values) (4,29). With the current development of engineered in vitro liver models offering increased hepatocyte life span, one can now extend the confidence in clearance predictions towards the low range, which is important for once-daily dosing regimens.…”

Section: Discussionmentioning

confidence: 99%

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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“…In recent years, considerable efforts were made to provide advanced hepatic in vitro systems mitigating the loss of DME activities to reliably predict human CL of metabolically stable compounds (Di and Obach, 2015;Hutzler et al, 2015). However, limited availability and large variability in DME expression and activity remain shortcomings of PHH, in particular for routine screening applications in drug discovery.…”

Section: Discussionmentioning

confidence: 99%

“…Primary hepatocyte suspensions are commonly preferred for the in vitro determination of intrinsic CL (CL int ) , representing a fully competent metabolic model (Blanchard et al, 2006;Hewitt et al, 2007) with an expression pattern of drug-metabolizing enzymes (DMEs) comparable to the liver of origin (Richert et al, 2006). However, suspended hepatocytes lose viability and metabolic activity within a few hours (Skett, 1994;Gebhardt et al, 2003), which frequently averts reliable and accurate CL int determination, in particular for slowly cleared compounds (Di and Obach, 2015;Hutzler, et al, 2015).…”

Section: Introductionmentioning

confidence: 99%

Upcyte Human Hepatocytes: a Potent In Vitro Tool for the Prediction of Hepatic Clearance of Metabolically Stable Compounds

Schaefer

Schänzle

Bischoff

et al. 2015

Drug Metabolism and Disposition

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In vitro models based on primary human hepatocytes (PHH) have been advanced for clearance (CL) prediction of metabolically stable compounds, representing state-of-the-art assay systems for drug discovery and development. Yet, limited cell availability and large interindividual variability of metabolic profiles remain shortcomings of PHH. Upcyte human hepatocytes (UHH) represent a novel hepatic cell system derived from PHH, exhibiting proliferative capacity for approximately 35 population doublings. UHH from three donors were evaluated during culture for up to 18 days, investigating relative mRNA expression and in situ enzyme activity of cytochrome P450s (P450s), UDP-glucuronosyltransferases, and sulfotransferases. Furthermore, UHH were used for predicting hepatic CL of 21 marketed low to intermediate CL drugs. In a typical experiment, expansion from 3.9 3 10 6 up to 8.5 3 10 7 cells was achieved during subculture.When maintained at confluence, transcripts of major P450s were expressed at donor-specific levels with sustained activities for the majority of isoforms, showing generally low CYP1A2 and high CYP2B6 activity levels. For donor 151-03, CL prediction based on depletion experiments resulted in an average fold error of 2.0, and 80% of compounds being predicted within twofold to in vivo CL for a subset of 10 low CL drugs. UHH showed sustained and consistent activity of drug-metabolizing enzymes (DME), resulting in highly reproducible CL prediction performance. In conclusion, UHH show promising potential as alternative to PHH for standardized in vitro applications in discovery research based on their stable, hepatocytelike DME phenotype and virtually unlimited cell availability.

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“…For example, there are differences in the in vitro test system (e.g. subcellular fraction, primary cells, cell lines, liver slices) and the ability to deal with fast or slowly cleared compounds (Brandon et al, 2003;Di and Obach, 2015).…”

Section: Objective 14 -Human Metabolic Stability/clearance Methodologymentioning

confidence: 99%

EURL ECVAM strategy for achieving 3Rs impact in the assessment of toxicokinetics and systemic toxicity

et al. 2015

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Addressing the Challenges of Low Clearance in Drug Research

Cited by 76 publications

References 20 publications

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

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

Upcyte Human Hepatocytes: a Potent In Vitro Tool for the Prediction of Hepatic Clearance of Metabolically Stable Compounds

EURL ECVAM strategy for achieving 3Rs impact in the assessment of toxicokinetics and systemic toxicity

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