Balancing receptor engagement and payload catalytic activity broadens the therapeutic window of immunotoxins

Immunotoxins (ITs) are potent targeted cancer therapeutics that combine tumor-selective receptor binding with efficient intracellular delivery and the cytotoxic enzymatic activity of bacterial toxins. They have shown impressive efficacy in hematologic malignancies, but dose-limiting toxicity remains a major barrier to broader clinical application, particularly in solid tumors. The exceptional potency of immunotoxin payloads contributes to off-tumor toxicity because even limited uptake by receptor-low cells can be sufficient to cause cell death.

To address this, the study systematically investigated how receptor-binding domain (RBD) affinity and avidity interact with catalytic payload activity to determine IT selectivity, using CD123-, HER3-, and HER2-targeted ITs.

Increasing receptor-binding affinity or avidity altered potency but provided limited control over selectivity between receptor-high and receptor-low cells. In contrast, attenuating catalytic activity produced profound gains in selectivity when payload potency was appropriately matched to receptor engagement. The optimal level of attenuation varied with receptor context and toxin scaffold, demonstrating that maximal selectivity emerges from balancing cellular delivery with catalytic activity rather than maximizing either property alone.

In vivo, payload attenuation increased the maximum tolerated dose of a HER2-targeted IT by 400-fold and enabled robust antitumor activity well below its toxicity limit, whereas the WT counterpart showed no detectable efficacy at tolerated doses.

Together, these findings uncover and establish payload activity as a tunable determinant of immunotoxin selectivity and provide a framework for matching receptor engagement with catalytic potency to develop safer and more broadly applicable targeted toxins.