Accelerating drug development in infectious diseases using zebrafish disease models supported by pharmacokinetic pharmacodynamic modeling as new approach methodology (NAM)
- 1Division of Systems Pharmacology and Pharmacy, Leiden Academic Centre for Drug Research, Leiden University, Leiden, the Netherlands
Abstract
The development of novel therapeutics for infectious diseases remains a global health priority. To accelerate the treatment development, innovative strategies through new approach methodology (NAM) are needed to bridge speed of in vitro with predictive power of in vivo studies, while reducing mammalian experiments. The zebrafish (Danio rerio), particularly the embryo/larva, has been established as a valuable non-mammalian in vivo model in biomedical research. We developed a standardized and streamlined workflow for the zebrafish as NAM, which consisted of 3 steps: drug selection and efficacy evaluation, internal exposure assessment, and PKPD modelling. Compounds with higher tolerated doses than minimum inhibitory concentration were selected. Drug efficacy was quantified through longitudinal individual fluorescence microscopy at baseline and 24 and 48h on treatment. Drug exposure was quantified in larval homogenates and exposure medium from 0-48h on treatment. The PKPD relationship was quantified by non-linear mixed effects modelling. For case study bedaquiline, PKPD was quantified using a one-compartment model with age-depending elimination, and an Emax concentration-response relationship on the delayed logistic bacterial growth function, with an EC50 of 26.5 g/mL and an Emax of 1.09-1.37. In the case of clarithromycin, in contrast, negligible internal exposure after waterborne treatment was observed, illustrating the risk of false negatives without internal exposure assessments. Bactericidal efficacy was confirmed by intravenous drug injections, showing a clear dose dependent antimycobacterial effect. The standardized zebrafish NAM workflow presented here facilitates the translation of drug efficacy to higher vertebrates, reducing rodent studies to confirmatory or replacing them completely, thus accelerating drug development.
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BibTeX
@misc{forncuni2026accelerating,
title = {{Accelerating drug development in infectious diseases using zebrafish disease models supported by pharmacokinetic pharmacodynamic modeling as new approach methodology (NAM)}},
author = {G. Forn-Cuni and B. van Lieshout and B. Koch and C. Villellas and S. Van Asten and D. Berlina and E. Lanckacker and B. Stoops and R. J. Vreeken and D. Roymans and E. H. J. Krekels and J. G. C. van Hasselt and H. P. Spaink and R. C. van Wijk},
howpublished = {bioRxiv},
year = {2026},
month = may,
doi = {10.64898/2026.05.29.728504},
url = {https://www.biorxiv.org/content/10.64898/2026.05.29.728504v2},
note = {CC BY},
}
RIS
TY - UNPB
TI - Accelerating drug development in infectious diseases using zebrafish disease models supported by pharmacokinetic pharmacodynamic modeling as new approach methodology (NAM)
AU - G. Forn-Cuni
AU - B. van Lieshout
AU - B. Koch
AU - C. Villellas
AU - S. Van Asten
AU - D. Berlina
AU - E. Lanckacker
AU - B. Stoops
AU - R. J. Vreeken
AU - D. Roymans
AU - E. H. J. Krekels
AU - J. G. C. van Hasselt
AU - H. P. Spaink
AU - R. C. van Wijk
PY - 2026
DA - 2026/05/29
PB - bioRxiv
DO - 10.64898/2026.05.29.728504
UR - https://www.biorxiv.org/content/10.64898/2026.05.29.728504v2
AB - The development of novel therapeutics for infectious diseases remains a global health priority. To accelerate the treatment development, innovative strategies through new approach methodology (NAM) are needed to bridge speed of in vitro with predictive power of in vivo studies, while reducing mammalian experiments. The zebrafish (Danio rerio), particularly the embryo/larva, has been established as a valuable non-mammalian in vivo model in biomedical research. We developed a standardized and streamlined workflow for the zebrafish as NAM, which consisted of 3 steps: drug selection and efficacy evaluation, internal exposure assessment, and PKPD modelling. Compounds with higher tolerated doses than minimum inhibitory concentration were selected. Drug efficacy was quantified through longitudinal individual fluorescence microscopy at baseline and 24 and 48h on treatment. Drug exposure was quantified in larval homogenates and exposure medium from 0-48h on treatment. The PKPD relationship was quantified by non-linear mixed effects modelling. For case study bedaquiline, PKPD was quantified using a one-compartment model with age-depending elimination, and an Emax concentration-response relationship on the delayed logistic bacterial growth function, with an EC50 of 26.5 g/mL and an Emax of 1.09-1.37. In the case of clarithromycin, in contrast, negligible internal exposure after waterborne treatment was observed, illustrating the risk of false negatives without internal exposure assessments. Bactericidal efficacy was confirmed by intravenous drug injections, showing a clear dose dependent antimycobacterial effect. The standardized zebrafish NAM workflow presented here facilitates the translation of drug efficacy to higher vertebrates, reducing rodent studies to confirmatory or replacing them completely, thus accelerating drug development.
ER -