Accelerating development of drug (combinations) against infectious diseases is a global priority. Cases of mycobacterial infections, such as tuberculosis (TB) and non-tuberculous mycobacterial (NTM) infections are stationary or even rising nationally and globally [1-3]. These infections are especially hard to treat, requiring extensive treatment durations with multiple antibiotics. Translational predictive models, predicting clinical efficacy (or lack thereof) for drug combinations have been successfully developed based on rodent and rabbit in vivo data [4-6]. At the same time, new approach methodologies (NAM) to mammalian in vivo experiments are necessary [7]. One example of NAM is the lower phylogenetically vertebrate zebrafish (Danio rerio), combined with predictive pharmacokinetic-pharmacodynamic (PKPD) modelling [8]. The zebrafish embryo/larva is optically transparent allowing for longitudinal, microscopy-based quantification of for example fluorescent pathogens during disease progression and treatment [9]. Benefiting from lessons-learnt in mammalian predictive models, we have successfully quantified both PK and PD in the zebrafish mycobacterial infection model and successfully predicted clinical efficacy in several phase 2a trials of cornerstone antibiotics isoniazid and bedaquiline. With recent possibilities of automation through robotics, the zebrafish NAM model supported by PKPD modelling, will improve our understanding of drug efficacy and exposure earlier, accelerating and de-risking development of the most promising therapeutics.
[1] World Health Organization Global tuberculosis report 2025. (Geneva, 2025); [2] RIVM, Kerncijfers tuberculose (2025); [3] Lange et al, Pathog Immun 10(2) (2025); [4] Ernest et al, Eur Respir J 62, 2300165 (2023); [5] Ernest et al, Nat Comm eprint (2026); [6] Strydom et al, Sci Transl Med 17, eadi4000 (2025); [7] U.S. Department of Health and Human Service, Food and Drug Administration Web announcement: FDA releases draft guidance alternatives animal testing drug development (2026); [8] Forn-Cuní et al, bioRxiv 10.64898/2026.05.29.728504v1 (2026); [9] Van Wijk et al Br J Pharmacol 177, 5518–5533 (2020)