Programmable DNA‐Peptide Hybrid Nanostructures for Potent Neutralization of Multiple Influenza a Virus Subtypes

ABSTRACT The continued emergence of antigenic drift and drug‐resistant viral strains highlights the need for antiviral strategies that deliver robust efficacy, broad subtype coverage, and minimal off‐target toxicity. We demonstrate a potent and broad‐spectrum strategy that employs hybrid biomaterials of Urumin (a host defense peptide) and a honeycomb (HC) DNA origami through spatially organized multivalent presentation for enhanced antiviral efficacy. Molecular dynamics simulations reveal that Urumin penetrates and destabilizes the hemagglutinin (HA) trimer core, disrupting influenza A viral (IAV) entry. Arranging Urumin in trimeric clusters on the HC enables potent multivalent binding to trimeric HAs on IAV, enhancing antiviral efficacy at nanomolar concentrations, ∼1000‐fold more effective than free Urumin. In vitro assays confirm HC‐Urumin outperforms free Urumin in blocking viral entry and preserving cell viability in more IAV subtypes. In vivo studies show that compared to free Urumin, HC‐Urumin treatment reduces disease severity, preserves physiological behavior, and decreases mortality in infected mice, while maintaining virus‐specific adaptive immune responses without altering humoral immunity. Our study offers an advanced and effective materials platform and strategy for broad‐spectrum, low‐dose intervention against human and animal IAVs, which can be adapted to combat other viruses by patterning corresponding host defense peptides on custom designed DNA nanostructures.