Fig. 5: PZ48 does not harm normal blood cells and is effective against ALL cells in vivo.

A PBMCs were treated with increasing doses (5 µM, 10 µM, 15 µM) of PZ48 for 24 h. Staining of cells was done with Annexin-V AF647 and FVD eFl780. The cells were analyzed using flow cytometry (n = 3, mean+SD; two-way ANOVA; n.s., not significant). Isolated subtypes of cells were defined: CD3-CD19+ as B cells; CD3+ as T cells; CD3-CD19-CD56+ as NK cells; CD3-CD19-CD1c+ as dendritic cells; CD3-CD19-CD14+ as monocytes; and CD3-CD14-CD19-CD56-CD11b+ as PMNs. B PBMCs were incubated with R848 (1 µg/ml) or Dynabeads™ Human T-Activator CD3/CD28 (5 µl/ml) for 24 h (control, DMSO treatment). Then, PBMCs were treated with increasing doses (5 µM, 10 µM, 15 µM) of PZ48 for 24 h. Staining of FVD eFl780 was analyzed using flow cytometry (n = 5, mean+SD; two-way ANOVA; n.s., not significant). T cells were defined as CD3 + , B cells were defined as CD3-CD19 + . C Model of the Danio rerio experiment. Created in BioRender. Mieland, A. (2025) https://BioRender.com/t85m878. D Waterfall plots demonstrating changes in tumor volume [%] for each individual zebrafish larvae engrafted with RS4-11 cells, from baseline (day 1 = start of the treatment) until day 3 after injection of such cells. Zebrafish larvae xenografts were treated with DMSO used as a solvent (n = 13 larvae; left) or 40 µM PZ48 (n = 11 larvae; right) for 48 h; each bar reflects one individual xenograft. Numbers indicate the percentages of early larvae with progressive disease (PD), stable disease (SD), and partial response (PR) in each treatment group on day 3.