Computational design of potent, broadly neutralizing anti-Nipah virus and Hendra virus miniproteins

The prototype members of the genus Henipavirus, Nipah virus (NiV) and Hendra virus (HeV), cause recurrent zoonotic spillovers with case fatality rates ranging from 40-90% in humans. Currently, there are no approved vaccines or therapeutics for use in humans. Neutralizing antibodies targeting the NiV/HeV F- or G-glycoproteins protect animals from lethal challenge and are a main correlate of protection. However, antibody-based formulations are expensive, typically requiring hospital admission for administration and cold-chain for storage and transportation. To address the lack of shelf-stable clinical countermeasures, we computationally designed thermostable miniproteins that cross-react with subnanomolar affinities with both NiV and HeV F and G glycoproteins and inhibit viral entry in vitro with potencies comparable to lead antibodies. Oligomerized forms of these miniproteins have enhanced potency relative to their monomeric building blocks and increase the barrier for emergence of escape mutants, establishing them as promising preclinical candidates against these deadly viruses.