Abstract

Viral infections kill millions yearly. Available antiviral drugs are virus-specific and active against a limited panel of human pathogens. There are broad-spectrum substances that prevent the first step of virus–cell interaction by mimicking heparan sulfate proteoglycans (HSPG), the highly conserved target of viral attachment ligands (VALs). The reversible binding mechanism prevents their use as a drug, because, upon dilution, the inhibition is lost. Known VALs are made of closely packed repeating units, but the aforementioned substances are able to bind only a few of them. We designed antiviral nanoparticles with long and flexible linkers mimicking HSPG, allowing for effective viral association with a binding that we simulate to be strong and multivalent to the VAL repeating units, generating forces (190 pN) that eventually lead to irreversible viral deformation. Virucidal assays, electron microscopy images, and molecular dynamics simulations support the proposed mechanism.  These particles show no cytotoxicity, and in vitro nanomolar irreversible activity against herpes simplex virus (HSV), human papilloma virus, respiratory syncytial virus (RSV), dengue and lenti virus. They are active ex vivo in human cervicovaginal histocultures infected by HSV-2 and in vivo in mice infected with RSV.

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References

  1. 1.

    et al. Global and regional mortality from 235 causes of death for 20 age groups in 1990 and 2010: a systematic analysis for the Global Burden of Disease Study 2010. Lancet (London, England) 380, 2095–2128 (2012).

  2. 2.

    Top 10 causes of death. WHO (2017).

  3. 3.

    Vaccines: past, present and future. Nat. Med. 11, S5–S11 (2005).

  4. 4.

    & Approved antiviral drugs over the past 50 years. Clin. Microbiol. Rev. 29, 695–747 (2016).

  5. 5.

    Strategies in the design of antiviral drugs. Nat. Rev. Drug Discov. 1, 13–25 (2002).

  6. 6.

    , & Cellular factors for resistance against antiretroviral agents. Antivir. Ther. 5, 181–185 (2000).

  7. 7.

    Heparan sulfate: anchor for viral intruders? Biochimie 83, 811–817 (2001).

  8. 8.

    et al. Highly sulfated K5 Escherichia coli polysaccharide derivatives inhibit respiratory syncytial virus infectivity in cell lines and human tracheal-bronchial histocultures. Antimicrob. Agents Chemother. 58, 4782–4794 (2014).

  9. 9.

    et al. Auto-associative heparin nanoassemblies: a biomimetic platform against the heparan sulfate-dependent viruses HSV-1, HSV-2, HPV-16 and RSV. Eur. J. Pharmaceutics Biopharmaceutics: Official Journal of Arbeitsgemeinschaft Für Pharmazeutische Verfahrenstechnik e.V 88, 275–282 (2014).

  10. 10.

    et al. Sulfated K5 Escherichia coli polysaccharide derivatives: a novel class of candidate antiviral microbicides. Pharmacol. Ther. 123, 310–322 (2009).

  11. 11.

    , , , & Inhibition of human metapneumovirus binding to heparan sulfate blocks infection in human lung cells and airway tissues. J. Virol. 90, 9237–9250 (2016).

  12. 12.

    et al. A haploid genetic screen identifies heparan sulfate proteoglycans supporting Rift Valley fever virus infection. J. Virol. 90, 1414–1423 (2015).

  13. 13.

    et al. The AGMA1 poly(amidoamine) inhibits the infectivity of herpes simplex virus in cell lines, in human cervicovaginal histocultures, and in vaginally infected mice. Biomaterials 85, 40–53 (2016).

  14. 14.

    et al. The agmatine-containing poly(amidoamine) polymer AGMA1 binds cell surface heparan sulfates and prevents attachment of mucosal human papillomaviruses. Antimicrob. Agents Chemother. 59, 5250–5259 (2015).

  15. 15.

    , , & Inhibition of HSV-1 attachment, entry, and cell-to-cell spread by functionalized multivalent gold nanoparticles. Small 6, 1044–1050 (2010).

  16. 16.

    et al. Polysulfonates derived from metal thiolate complexes as inhibitors of HIV-1 and various other enveloped viruses in vitro. Antivir. Chem. Chemother. 13, 185–195 (2002).

  17. 17.

    et al. Inhibition of HIV fusion with multivalent gold nanoparticles. J. Am. Chem. Soc. 130, 6896–6897 (2008).

  18. 18.

    et al. Candidate sulfonated and sulfated topical microbicides: comparison of anti-human immunodeficiency virus activities and mechanisms of action. Antimicrob. Agents Chemother. 49, 3607–3615 (2005).

  19. 19.

    et al. PRO2000 vaginal gel for prevention of HIV-1 infection (Microbicides Development Programme 301): a phase 3, randomised, double-blind, parallel-group trial. Lancet 376, 1329–1337 (2010).

  20. 20.

    , & The rise and fall of polyanionic inhibitors of the human immunodeficiency virus type 1. Antivir. Res. 90, 168–182 (2011).

  21. 21.

    et al. Lack of effectiveness of cellulose sulfate gel for the prevention of vaginal HIV transmission. New Engl. J. Med. 359, 463–472 (2008).

  22. 22.

    , , , & Virucidal activity of a GT-rich oligonucleotide against herpes simplex virus mediated by glycoprotein B. J. Virol. 80, 4740–4747 (2006).

  23. 23.

    et al. Cell-free HIV-1 virucidal action by modified peptide triazole inhibitors of Env gp120. ChemMedChem 6, 1335–1339 (2011).

  24. 24.

    et al. Viral inhibition mechanism mediated by surface-modified silica nanoparticles. ACS Appl. Mater. Interfaces 8, 16564–16572 (2016).

  25. 25.

    et al. Antiviral properties of polymeric aziridine- and biguanide-modified core-shell magnetic nanoparticles. Langmuir 28, 4548–4558 (2012).

  26. 26.

    et al. Antiviral activity of gold/copper sulfide core/shell nanoparticles against human norovirus virus-like particles. PLoS ONE 10, e0141050 (2015).

  27. 27.

    , , & Silver nanoparticles are broad-spectrum bactericidal and virucidal compounds. J. Nanobiotechnology 9, 30 (2011).

  28. 28.

    , , , & Inhibitory effects of silver nanoparticles against adenovirus type 3 in vitro. J. Virol. Methods 193, 470–477 (2013).

  29. 29.

    et al. Effects of several virucidal agents on inactivation of influenza, Newcastle disease, and avian infectious bronchitis viruses in the allantoic fluid of chicken eggs. Jpn. J. Infect. Dis. 60, 342–346 (2007).

  30. 30.

    , & The effect of interactions on the cellular uptake of nanoparticles. Phys. Biol. 8, 046002 (2011).

  31. 31.

    & Vesicles in contact with nanoparticles and colloids. Europhys. Lett. 43, 219–225 (1998).

  32. 32.

    et al. A general mechanism for intracellular toxicity of metal-containing nanoparticles. Nanoscale 6, 7052–7061 (2014).

  33. 33.

    , , & Colloidal stability of self-assembled mono layer-coated gold nanoparticles: the effects of surface compositional and structural heterogeneity. Langmuir 29, 11560–11566 (2013).

  34. 34.

    , , & Protein-nanoparticle interactions: the effects of surface compositional and structural heterogeneity are scale dependent. Nanoscale 5, 6928–6935 (2013).

  35. 35.

    , , , & Effects of surface compositional and structural heterogeneity on nanoparticle-protein interactions: different protein configurations. ACS Nano 8, 5402–5412 (2014).

  36. 36.

    , & Pathogen inhibition by multivalent ligand architectures. J. Am. Chem. Soc. 138, 8654–8666 (2016).

  37. 37.

    et al. Structural basis of oligosaccharide receptor recognition by human papillomavirus. J. Biol. Chem. 286, 2617–2624 (2011).

  38. 38.

    et al. Surface-exposed amino acid residues of HPV16 l1 protein mediating interaction with cell surface heparan sulfate. J. Biol. Chem. 282, 27913–27922 (2007).

  39. 39.

    et al. Atomically precise organomimetic cluster nanomolecules assembled via Perfluoroaryl-Thiol SNAr Chemistry. Nat. Chem. 9, 333–340 (2016).

  40. 40.

    & 1-Anilino-8-naphthalene sulfonate anion-protein binding depends primarily on ion pair formation. Biophys. J. 74, 422–429 (1998).

  41. 41.

    et al. The complex nature of calcium cation interactions with phospholipid bilayers. Sci. Rep. 6, 38035 (2016).

  42. 42.

    et al. Surface-structure-regulated cell-membrane penetration by monolayer-protected nanoparticles. Nat. Mater. 7, 588–595 (2008).

  43. 43.

    et al. Visualizing the replication of respiratory syncytial virus in cells and in living mice. Nat. Commun. 5, 5104 (2014).

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Acknowledgements

F.S. and his laboratory were supported in part by the Swiss National Science Foundation NRP 64 grant, and by the NCCR on bio-inspired materials. D.L. was supported by a grant from University of Turin (ex 60%). J.H. and J.W. were supported by a research grant from the Ministry of Education, Youth and Sports of the Czech Republic (LK11207). C.T., L.K. and F.S. were supported by the Leenaards Foundation. P.K. was supported by the NSF DMR-1506886 grant. L.V. was supported by startup funding from UTEP. M.G. and R.L. thank the MIMA2 platform for access to the IVIS 200, which was financed by the Ile de France region (SESAME). M.M. thanks R. C. Guerrero-Ferreira for the tomogram acquisition. P.A. was supported by funding from the European Union Horizon, H2020 Nanofacturing, under grant agreement 646364.

Author information

Author notes

    • Valeria Cagno
    •  & Patrizia Andreozzi

    These authors contributed equally to this work.

Affiliations

  1. Dipartimento di Scienze Cliniche e Biologiche, Univerisità degli Studi di Torino, Orbassano, Italy

    • Valeria Cagno
    • , Manuela Donalisio
    •  & David Lembo
  2. Institute of Materials, Ecole Polytechnique Fédérale de Lausanne (EPFL), Lausanne, Switzerland

    • Valeria Cagno
    • , Paulo Jacob Silva
    • , Marie Mueller
    • , Samuel T. Jones
    • , Emma-Rose Janeček
    • , Ahmet Bekdemir
    •  & Francesco Stellacci
  3. Faculty of Medicine of Geneva, Department of Microbiology and Molecular medicine, Geneva, Switzerland

    • Valeria Cagno
    •  & Caroline Tapparel
  4. IFOM - FIRC Institute of Molecular Oncology, IFOM-IEO Campus, Milan, Italy

    • Patrizia Andreozzi
    •  & Chiara Martinelli
  5. CIC biomaGUNE Soft Matter Nanotechnology Group San Sebastian-Donostia, 20014 Donastia San Sebastián, Spain

    • Patrizia Andreozzi
  6. Fondazione Centro Europeo Nanomedicina (CEN), Milan, Italy

    • Marco D’Alicarnasso
  7. VIM, INRA, Université Paris-Saclay, Jouy-en-Josas, France

    • Marie Galloux
    • , Ronan Le Goffic
    •  & Jean-Francois Eleouet
  8. Jones Lab, School of Materials, University of Manchester, Oxford Road, Manchester M13 9PL, UK

    • Samuel T. Jones
  9. Istituto per la Protezione Sostenibile delle Piante, CNR, Torino, Italy

    • Marta Vallino
  10. Institute of Organic Chemistry and Biochemistry of the Czech Academy of Sciences, Prague, Czech Republic

    • Jan Hodek
    •  & Jan Weber
  11. Department of Chemistry, University of Illinois at Chicago, Chicago, Illinois 60607, USA

    • Soumyo Sen
    • , Yanxiao Han
    •  & Petr Král
  12. Fondazione IRCCS Istituto Neurologico “Carlo Besta”, IFOM-IEO Campus, Milan, Italy

    • Barbara Sanavio
    •  & Silke Krol
  13. UMR INSERM U1173 I2, UFR des Sciences de la Santé Simone Veil—UVSQ, Montigny-Le-Bretonneux, France

    • Marie-Anne Rameix Welti
  14. AP-HP, Laboratoire de Microbiologie, Hôpital Ambroise Paré, 92104 Boulogne-Billancourt, France

    • Marie-Anne Rameix Welti
  15. Geneva University Hospitals, Infectious Diseases Divisions, Geneva, Switzerland

    • Laurent Kaiser
    •  & Caroline Tapparel
  16. Department of Chemistry, University of Texas at El Paso, El Paso, Texas 79968, USA

    • Lela Vukovic
  17. Department of Physics and Department of Biopharmaceutical Sciences, University of Illinois at Chicago, Chicago, Illinois 60607, USA

    • Petr Král
  18. IRCCS Istituto Tumori “Giovanni Paolo II”, Bari, Italy

    • Silke Krol
  19. Interfaculty Bioengineering Institute, Ecole Polytechnique Fédérale de Lausanne (EPFL), Lausanne, Switzerland

    • Francesco Stellacci

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Contributions

V.C. was responsible for all activities involving HSV2, HPV and RSV under the supervision of D.L. and EpiVaginal experiments under the supervision of C.T. and L.K. P.A. M.D. and C.M. were responsible for all testing with VSV-LV-G under the direction of S.K. P.J.S. was responsible for NP and ligand synthesis. M.M. was responsible for all cryo-TEM. S.T.J. was responsible for iron oxide NP synthesis. M.G. and R.L. were responsible for the in vivo experiments, R.W.M. and J.F.E. engineered the RSV-Luc used for in vivo experiments. M.V. was responsible for stained TEM imaging of the viruses. J.H. and J.W. conducted all testing with DENV-2. S.S. and Y.H. were responsible for molecular dynamics simulations under the direction of P.K., and L.V. E.R.J. and S.T.J. synthesized MUP-NPs. A.B. synthesized MES-NPs. B.S. synthesized EG2OH-NPs. M.D. was responsible for HSV-1 and HSV-2 and dose response experiments. F.S. and S.K. first conceived the experiments, F.S. and D.L. developed the interpretation of the experiments. F.S., D.L., V.C. and S.T.J. wrote the paper.

Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to David Lembo or Francesco Stellacci.

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DOI

https://doi.org/10.1038/nmat5053

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