title: Pharmacokinetic Modeling of Paracetamol Uptake and Clearance in Zebrafish Larvae: Expanding the Allometric Scale in Vertebrates with Five Orders of Magnitude.
abstract: Zebrafish larvae (Danio rerio) are increasingly used to translate findings regarding drug efficacy and safety from in vitro-based assays to vertebrate species, including humans. However, the limited understanding of drug exposure in this species hampers its implementation in translational research. Using paracetamol as a paradigm compound, we present a novel method to characterize pharmacokinetic processes in zebrafish larvae, by combining sensitive bioanalytical methods and nonlinear mixed effects modeling. The developed method allowed quantification of paracetamol and its two major metabolites, paracetamol-sulfate and paracetamol-glucuronide in pooled samples of five lysed zebrafish larvae of 3 days post-fertilization. Paracetamol drug uptake was quantified to be 0.289 pmole/min and paracetamol clearance was quantified to be 1.7% of the total value of the larvae. With an average volume determined to be 0.290 μL, this yields an absolute clearance of 2.96 × 107L/h, which scales reasonably well with clearance rates in higher vertebrates. The developed methodology will improve the success rate of drug screens in zebrafish larvae and the translation potential of findings, by allowing the establishment of accurate exposure profiles and thereby also the establishment of concentration-effect relationships.
authors.affiliation: 1 Division of Analytical Biosciences, Systems Pharmacology Cluster, Leiden Academic Centre for Drug Research, Leiden University , Leiden, The Netherlands .
authors.forename: Vasudev
authors.initials: V
authors.lastname: Kantae
authors.affiliation: 2 Division of Pharmacology, Systems Pharmacology Cluster, Leiden Academic Centre for Drug Research, Leiden University , Leiden, The Netherlands .
authors.forename: Elke H J
authors.initials: EH
authors.lastname: Krekels
authors.affiliation: 3 IBL, Institute of Biology, Leiden University , Leiden, the Netherlands .
authors.forename: Anita
authors.initials: A
authors.lastname: Ordas
authors.affiliation: 1 Division of Analytical Biosciences, Systems Pharmacology Cluster, Leiden Academic Centre for Drug Research, Leiden University , Leiden, The Netherlands .|4 Science and Technology Faculty, Analytical Chemistry Department, University of the Basque Country/EHU , Bilbao, Spain .
authors.forename: Oskar
authors.initials: O
authors.lastname: González
authors.affiliation: 2 Division of Pharmacology, Systems Pharmacology Cluster, Leiden Academic Centre for Drug Research, Leiden University , Leiden, The Netherlands .
authors.forename: Rob C
authors.initials: RC
authors.lastname: van Wijk
authors.affiliation: 1 Division of Analytical Biosciences, Systems Pharmacology Cluster, Leiden Academic Centre for Drug Research, Leiden University , Leiden, The Netherlands .
authors.forename: Amy C
authors.initials: AC
authors.lastname: Harms
authors.affiliation: 5 ZF-screens B.V. , Leiden, The Netherlands .
authors.forename: Peter I
authors.initials: PI
authors.lastname: Racz
authors.affiliation: 2 Division of Pharmacology, Systems Pharmacology Cluster, Leiden Academic Centre for Drug Research, Leiden University , Leiden, The Netherlands .
authors.forename: Piet H
authors.initials: PH
authors.lastname: van der Graaf
authors.affiliation: 3 IBL, Institute of Biology, Leiden University , Leiden, the Netherlands .
authors.forename: Herman P
authors.initials: HP
authors.lastname: Spaink
authors.affiliation: 1 Division of Analytical Biosciences, Systems Pharmacology Cluster, Leiden Academic Centre for Drug Research, Leiden University , Leiden, The Netherlands .
authors.forename: Thomas
authors.initials: T
authors.lastname: Hankemeier
mesh_terms: D000082:Acetaminophen / Q000493:pharmacokinetics*; D018712:Analgesics, Non-Narcotic / Q000493:pharmacokinetics*; D000818:Animals; D002853:Chromatography, Liquid; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D013058:Mass Spectrometry; D008962:Models, Theoretical; D017711:Nonlinear Dynamics; D015027:Zebrafish / Q000378:metabolism*
chemical_list: D018712:Analgesics, Non-Narcotic; D000082:Acetaminophen
keywords: allometry; drug discovery; metabolism; paracetamol; pharmacokinetics; zebrafish