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Crystal structures of oseltamivir-resistant influenza virus neuraminidase mutants

Nature volume 453, pages 1258–1261 (2008)Cite this article

Abstract

The potential impact of pandemic influenza makes effective measures to limit the spread and morbidity of virus infection a public health priority. Antiviral drugs are seen as essential requirements for control of initial influenza outbreaks caused by a new virus, and in pre-pandemic plans there is a heavy reliance on drug stockpiles. The principal target for these drugs is a virus surface glycoprotein, neuraminidase, which facilitates the release of nascent virus and thus the spread of infection. Oseltamivir (Tamiflu) and zanamivir (Relenza) are two currently used neuraminidase inhibitors that were developed using knowledge of the enzyme structure1,2. It has been proposed that the closer such inhibitors resemble the natural substrate, the less likely they are to select drug-resistant mutant viruses that retain viability3. However, there have been reports of drug-resistant mutant selection in vitro4 and from infected humans5,6. We report here the enzymatic properties and crystal structures of neuraminidase mutants from H5N1-infected patients that explain the molecular basis of resistance. Our results show that these mutants are resistant to oseltamivir but still strongly inhibited by zanamivir owing to an altered hydrophobic pocket in the active site of the enzyme required for oseltamivir binding. Together with recent reports of the viability and pathogenesis of H5N1 (ref. 7) and H1N1 (ref. 8) viruses with neuraminidases carrying these mutations, our results indicate that it would be prudent for pandemic stockpiles of oseltamivir to be augmented by additional antiviral drugs, including zanamivir.

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Figure 1: Neuraminidase activity monitored using a fluorescent assay.
Figure 2: Structure of N1 neuraminidase complexes.

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Protein Data Bank

Data deposits

Structural data have been deposited with the Protein Data Bank with accession codes 3CL0 (His274Tyr–oseltamivir), 3CKZ (His274Tyr–zanamivir) and 3CL2 (Asn294Ser–oseltamivir).

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Acknowledgements

Work at NIMR was funded by the Medical Research Council (UK). This work was also supported in part by the EU FP6 Programme VIRGIL, contract number 503359. R.J.R. thanks the Scottish Funding Council for financial support.

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Authors and Affiliations

  1. MRC-National Institute for Medical Research, Mill Hill, London NW7 1AA, UK ,

    Patrick J. Collins, Lesley F. Haire, Yi Pu Lin, Junfeng Liu, Philip A. Walker, John J. Skehel, Stephen R. Martin, Alan J. Hay & Steven J. Gamblin

  2. Interdisciplinary Centre for Human and Avian Influenza Research, School of Biology, University of St Andrews, Fife KY16 9ST, UK

    Rupert J. Russell

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  1. Patrick J. Collins
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Correspondence to Steven J. Gamblin.

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The file contains Supplementary Notes, Supplementary Figures S1-S2 with Legends and Supplementary Table 1 and additional references. (PDF 219 kb)

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Collins, P., Haire, L., Lin, Y. et al. Crystal structures of oseltamivir-resistant influenza virus neuraminidase mutants. Nature 453, 1258–1261 (2008). https://doi.org/10.1038/nature06956

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