Agriculture and Food Chemistry

Amantadine Variants Activity Against Multiple Influenza A Viruses

Authors

  • Μarianna Stampolaki Laboratory of Medicinal Chemistry, Section of Pharmaceutical Chemistry, Department of Pharmacy, National and Kapodistrian University of Athens, Panepistimiopolis-Zografou, 15771, Athens, Greece ,
  • Christina Tzitzoglaki Laboratory of Medicinal Chemistry, Section of Pharmaceutical Chemistry, Department of Pharmacy, National and Kapodistrian University of Athens, Panepistimiopolis-Zografou, 15771, Athens, Greece ,
  • Christos Liolios Laboratory of Medicinal Chemistry, Section of Pharmaceutical Chemistry, Department of Pharmacy, National and Kapodistrian University of Athens, Panepistimiopolis-Zografou, 15771, Athens, Greece ,
  • Anja Hoffmann Jena University Hospital, Department of Medical Microbiology, Section Experimental Virology, CMB Building, R. 443, Hans Knoell Str. 2, D-07745 Jena, Germany ,
  • Brent Johnson Department of Microbiology and Molecular Biology, Brigham Young University, Provo, UT 84602, United States ,
  • Ioannis Stylianakis Laboratory of Medicinal Chemistry, Section of Pharmaceutical Chemistry, Department of Pharmacy, National and Kapodistrian University of Athens, Panepistimiopolis-Zografou, 15771, Athens, Greece ,
  • Roland Zell Jena University Hospital, Department of Medical Microbiology, Section Experimental Virology, CMB Building, R. 443, Hans Knoell Str. 2, D-07745 Jena, Germany ,
  • Andreea Turcu Laboratori de Química Farmacèutica (Unitat Associada al CSIC), Facultat de Farmàcia i Ciències de l’Alimentació, and Institute of Biomedicine (IBUB), Universitat de Barcelona, Av. Joan XXIII, 27-31, Barcelona, E-08028, Spain ,
  • Patrick Schmerer Jena University Hospital, Department of Medical Microbiology, Section Experimental Virology, CMB Building, R. 443, Hans Knoell Str. 2, D-07745 Jena, Germany ,
  • Kristin Döring Jena University Hospital, Department of Medical Microbiology, Section Experimental Virology, CMB Building, R. 443, Hans Knoell Str. 2, D-07745 Jena, Germany ,
  • Chunlong Ma Department of Pharmacology and Toxicology, College of Pharmacy, University of Arizona, Tucson, Arizona 85721, United States ,
  • Ivi Theodosia Antoniadou Laboratory of Pharmaceutical Technology, Section of Pharmaceutical Technology, Department of Pharmacy, National and Kapodistrian University of Athens, Panepistimiopolis-Zografou, 15771, Athens, Greece; Present address: Biomedical Research Foundation of the Academy of Athens, 4 Soranou Ephessiou Str., 115 27 Athens, Greece ,
  • David Fedida Department of Anesthesiology, Pharmacology and Therapeutics, University of British Columbia, Vancouver, British Columbia, V6T 1Z3, Canada ,
  • Fransesc Sureda Department of Biochemistry and Biotechnology, Faculty of Medicine and Life Science, University Rovira i Virgili, Reus, Spain ,
  • Stephen Wharton Crick Worldwide Influenza Centre, The Francis Crick Institute, 1 Midland Road, London NW1 1AT, UK ,
  • Jun Wang 5 Department of Pharmacology and Toxicology, College of Pharmacy, University of Arizona, Tucson, Arizona 85721, United States; Present address: Department of Medicinal Chemistry, Ernest Mario School of Pharmacy Rutgers, The State University of New Jersey, 160 Frelinghuysen Road Piscataway, NJ 08854-8020 ,
  • Santiago Vázquez Laboratori de Química Farmacèutica (Unitat Associada al CSIC), Facultat de Farmàcia i Ciències de l’Alimentació, and Institute of Biomedicine (IBUB), Universitat de Barcelona, Av. Joan XXIII, 27-31, Barcelona, E-08028, Spain ,
  • Michaela Schmidtke Jena University Hospital, Department of Medical Microbiology, Section Experimental Virology, CMB Building, R. 443, Hans Knoell Str. 2, D-07745 Jena, Germany ,
  • Antonios Kolocouris Laboratory of Medicinal Chemistry, Section of Pharmaceutical Chemistry, Department of Pharmacy, National and Kapodistrian University of Athens, Panepistimiopolis-Zografou, 15771, Athens, Greece

Abstract

Future pandemic influenza necessitates the development of new drugs against the current circulating, amantadine and rimantadine drugs resistant, influenza A M2 S31N viruses. The possibility of an antigenic shift to M2 S31 necessitates ranking the biological activities of amantadine variants. Several amantadine variants have been tested by different laboratories, but various M2 wild type influenza A strains have been used with different sensitivity against amantadine and the unambiguous comparison between potencies is not straightforward. Here, we compared the anti-influenza activities of 57 synthetic amantadine variants against influenza A WSN/33 viruses with amantadine-sensitive M2 WT, with a range of over three digits providing a reference set of potencies for structure-activity relationships, and amantadine-resistant M2 S31N proteins (and observed no potent compounds). 17 compounds were selected and tested against M2 L26F, V27A, A30T, G34E viruses. We tested few reference compounds using electrophysiology and explored point mutations which both showed that M2 is the target of potent antiviral potency against the M2 WT, L26F, V27A viruses. Major findings are: (a) Several amantadine variants from Kolocouris group block only M2 WT and M2 L26F-mediated proton current and the corresponding viruses replication. (b) A compound from Vazquez’s group is a triple blocker of M2 WT, L26F, V27A channels and viruses replication. (c) A compound from Vazquez’s group blocks only M2 L26 channel and virus replication. (d) Several compounds from Kolocouris group have potent activity against several influenza A M2 WT and three M2 S31N viruses, eg. the pandemic A/H1N1/California/07/2009 (H1N1pdm09) or A/H1N1/PuertoRico/08/1934 without blocking M2 S31N. The compounds and their cocktails while not to be more toxic than amantadine might be useful for re-purposing of amantadine class of drugs in the case (i) of the prevalence of M2 L26F and or M2 V27A strains (ii) of an antigenic shift of the virus to M2 WT and (iii) because they inhibited a broad panel of M2 WT and M2 S31N viruses including the H1N1pdm09). (d) We showed that the mechanism of antiviral activity against A/California/07/2009 or A/PR/08/1934 and possibly also M2 WT viruses compared to WSN/33 viruses is not due to inhibition of an early stage of virus infection or a late stage of M2 channel function during endocytosis or inhibition of HA binding to host cells or a different pH for HA fusion or a lysosomotropic effect.

Content

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Supplementary material

Thumbnail image of SI_Kolocouris.pdf
Amantadine Variants Activity Against Multiple Influenza A Viruses
Synthetic chemistry methods and experimental methods for chemical synthesis and experimental methods for biological evaluation and neurotoxicity data, sequences alignments of M2 proteins from different viruses and SI references