COMMUNICATION
Attaching Zanamivir to a Polymer Markedly Enhances Its Activity Against Drug-resistant Strains of Influenza a Virus

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ABSTRACT

Effects of the commercial drug zanamivir (Relenza™) covalently attached to poly-l-glutamine on the infectivity of influenza A viruses are examined using the plaque reduction assay and binding affinity to viral neuraminidase (NA). These multivalent drug conjugates exhibit (i) up to a 20,000-fold improvement in anti-influenza potency compared with the zanamivir parent against human and avian viral strains, including both wild-type and drug-resistant mutants, and (ii) superior neuraminidase (NA) inhibition constants, especially for the mutants. These findings provide a basis for exploring polymer-attached inhibitors as more efficacious therapeutics, particularly against drug-resistant influenza strains.

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INTRODUCTION

In addition to yearly outbreaks affecting hundreds of millions of people worldwide,1 influenza pandemics of animal–human reassortment strains are increasingly common. Influenza infection is somewhat mitigated by prompt treatment with the neuraminidase (NA) inhibitors, oseltamivir (Tamiflu™), and zanamivir (Relenza™).1 Sialic acid cleavage from glycoproteins and glycolipids catalyzed by NA promotes both initiation of infection and multicycle infection by preventing viral aggregation, cleaving

MATERIALS AND METHODS

Poly-l-glutamic acid (molecular weight = 50,000–100,000 Da) and all other chemicals, biochemicals, and solvents were purchased from Sigma-Aldrich Chemical Co (St. Louis, Missouri, USA). Influenza viruses [A/Wuhan/359/95 (H3N2) and A/turkey/MN/833/80 (H4N2), as well as their drug-resistant mutants] were obtained from the Centers for Disease Control and Prevention (Atlanta, Georgia, USA). Madin-Darby canine kidney (MDCK) cells were purchased from the ATCC. Zanamivir (1, Fig. 1) was obtained from

RESULTS AND DISCUSSION

To explore the benefits of multivalency10 (i.e., in the present case, covalent attachment of multiple copies of zanamivir to the same polymeric chain) with respect to drug-resistant strains, herein we have selected three influenza A viruses carrying NA mutations at position 119: two in vitro selected zanamivir-resistant16 avian A/turkey/MN/833/80 (turkey/MN; H4N2) E119D and E119G mutants and a clinically isolated oseltamivir-resistant A/Wuhan/359/95 (Wuhan; H3N2) E119V mutant, which is

ACKNOWLEDGEMENTS

This work was partly supported by National Institutes of Health grant U01-AI074443. L.A.d.C. was supported by a postdoctoral fellowship from Fundación Ramón Areces of Spain.

REFERENCES (22)

  • D.M. Andrews et al.

    Synthesis and influenza sialidase inhibitory activity of analogues of 4-guanidino-Neu5Ac2en (zanamivir) modified in the glycerol side-chain

    Eur J Med Chem

    (1999)
  • M. Luscher-Mattlie

    Influenza chemotherapy: A review of the present state of art and of new drugs in development

    Arch Virol

    (2000)
  • O. Greengard et al.

    The anti-influenza virus agent 4-GU-DANA (zanamivir) inhibits cell fusion mediated by human parainfluenza virus and influenza virus HA

    J Virol

    (2000)
  • C. Liu et al.

    Influenza type A virus neuraminidase does not play a role in viral entry, replication, assembly, or budding

    J Virol

    (1995)
  • M.N. Matrosovich et al.

    Neuraminidase is important for the initiation of influenza virus infection in human airway epithelium

    J Virol

    (2004)
  • A. Moscona

    Oseltamivir resistance—disabling our influenza defenses

    N Engl J Med

    (2005)
  • V.P. Mishin et al.

    Susceptibilities of antiviral-resistant influenza viruses to novel neuraminidase inhibitors

    Antimicrob Agents Chemother

    (2005)
  • A.T. García-Sosa et al.

    Design of multi-binding-site inhibitors, ligand efficiency, and consensus screening of avian influenza H5N1 wild-type neuraminidase and of the oseltamivir-resistant H274Y variant

    J Chem Inf Model

    (2008)
  • K. Koyama et al.

    CS-8958, a prodrug of the novel neuraminidase inhibitor R-125489, demonstrates a favorable long-retention profile in the mouse respiratory tract

    Antimicrob Agents Chemother

    (2009)
  • M. Yamashita et al.

    CS-8959, a prodrug of the new neuraminidase inhibitor R-125489, shows long-acting anti-influenza virus activity

    Antimicrob Agents Chemother

    (2009)
  • M. Mammen et al.

    Polyvalent interactions in biological systems: Implications for design and use of multivalent ligands and inhibitors

    Angew Chem Int Ed

    (1998)
  • Cited by (0)

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