Binding mechanism of oseltamivir and influenza neuraminidase suggests perspectives for the design of new anti-influenza drugs

Author summaryInfluenza virus neuraminidase (NA), a viral membrane glycoprotein, plays an important role in the interactions with host cell surface receptors. The emergence and spread of influenza mutants resistant to neuraminidase inhibitors (NAIs), such as oseltamivir, has been of great concern. Despite many improvements to NAIs, no new first-line NAIs are currently in clinical use. Although there have been previous molecular dynamics simulation studies on the binding and dissociation process of oseltamivir-NA, we discovered new binding pathways and states of oseltamivir through larger-scale simulations and more systematic analysis, which may provide new ideas for the improvement of oseltamivir and even a series of NAIs. In our study, we strongly demonstrate that a detailed understanding of the drug-receptor association process is of fundamental importance for drug design and provide methodological references for the improvement of other drugs. Oseltamivir is a widely used influenza virus neuraminidase (NA) inhibitor that prevents the release of new virus particles from host cells. However, oseltamivir-resistant strains have emerged, but effective drugs against them have not yet been developed. Elucidating the binding mechanisms between NA and oseltamivir may provide valuable information for the design of new drugs against NA mutants resistant to oseltamivir. Here, we conducted large-scale (353.4 mu s) free-binding molecular dynamics simulations, together with a Markov State Model and an importance-sampling algorithm, to reveal the binding process of oseltamivir and NA. Ten metastable states and five major binding pathways were identified that validated and complemented previously discovered binding pathways, including the hypothesis that oseltamivir can be transferred from the secondary sialic acid binding site to the catalytic site. The discovery of multiple new metastable states, especially the stable bound state containing a water-mediated hydrogen bond between Arg118 and oseltamivir, may provide new insights into the improvement of NA inhibitors. We anticipated the findings presented here will facilitate the development of drugs capable of combating NA mutations.