PUBLICATION
Mechanistic and quantitative understanding of pharmacokinetics in zebrafish larvae through nanoscale blood sampling and metabolite modelling of paracetamol
- Authors
- van Wijk, R.C., Krekels, E.H., Kantae, V., Ordas, A., Kreling, T., Harms, A.C., Hankemeier, T., Spaink, H.P., van der Graaf, P.H.
- ID
- ZDB-PUB-190803-5
- Date
- 2019
- Source
- The Journal of pharmacology and experimental therapeutics 371(1): 15-24 (Journal)
- Registered Authors
- Spaink, Herman P.
- Keywords
- acetaminophen, animal models, computational models, drug discovery, drug disposition, glucuronidation, metabolite kinetics, pharmacokinetics, phase II drug metabolism, sulfation
- MeSH Terms
-
- Absorption, Physiological
- Acetaminophen/analogs & derivatives
- Acetaminophen/blood*
- Acetaminophen/pharmacokinetics
- Analgesics, Non-Narcotic/blood*
- Analgesics, Non-Narcotic/pharmacokinetics
- Animals
- Drug Evaluation, Preclinical/methods
- Drug Evaluation, Preclinical/standards
- Larva/metabolism
- Metabolic Clearance Rate
- Sensitivity and Specificity
- Tissue Distribution
- Zebrafish
- PubMed
- 31371482 Full text @ J. Pharmacol. Exp. Ther.
Citation
van Wijk, R.C., Krekels, E.H., Kantae, V., Ordas, A., Kreling, T., Harms, A.C., Hankemeier, T., Spaink, H.P., van der Graaf, P.H. (2019) Mechanistic and quantitative understanding of pharmacokinetics in zebrafish larvae through nanoscale blood sampling and metabolite modelling of paracetamol. The Journal of pharmacology and experimental therapeutics. 371(1):15-24.
Abstract
Zebrafish larvae are increasingly used for pharmacological research, but internal drug exposure is often not measured. Understanding pharmacokinetics is necessary for reliable translation of pharmacological results to higher vertebrates, including humans. Quantification of drug clearance and distribution requires measurements of blood concentrations. Additionally, measuring drug metabolites is of importance to understand clearance in this model organism mechanistically. We therefore mechanistically study and quantify pharmacokinetics in zebrafish larvae, and compare this to higher vertebrates, using paracetamol (acetaminophen) as paradigm compound. A method was developed to sample blood from zebrafish larvae at five days post fertilization. Blood concentrations of paracetamol and its major metabolites, paracetamol-glucuronide and paracetamol-sulphate, were measured. Blood concentration data were combined with measured amounts in larval homogenates and excreted amounts and simultaneously analysed through non-linear mixed effects modelling, quantifying absolute clearance and distribution volume. Blood sampling from zebrafish larvae was most successful from the posterior cardinal vein with median volume (interquartile range) of 1.12 (0.676-1.66) nL per blood sample. Samples were pooled (n=15-35) to reach measurable levels. Paracetamol blood concentrations at steady state were only 10% of the external paracetamol concentration. Paracetamol-sulphate was the major metabolite and its formation was quantified using a time-dependent metabolic formation rate. Absolute clearance and distribution volume correlated well to reported values in higher vertebrates, including humans. Based on blood concentrations and advanced data analysis, the mechanistic and quantitative understanding of paracetamol pharmacokinetics in zebrafish larvae has been established. This will improve the translational value of this vertebrate model organism in drug discovery and development. SIGNIFICANCE STATEMENT: In early phases of drug development, new compounds are increasingly screened in zebrafish larvae, but the internal drug exposure is often not taken into consideration. We developed innovative experimental and computational methods, including a blood sampling technique, to measure the paradigm drug paracetamol (acetaminophen) and its major metabolites and quantify pharmacokinetics (absorption, distribution, elimination) in zebrafish larvae of five days post fertilization with a total volume of only 300 nL. These parameter values were scaled to higher vertebrates, including humans.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping