Skip to main content
SpringerLink
Log in

Synthesis and Neuraminidase Inhibitory Activity of Sialic Acid Analogues with Fluoro, Phosphono, and Sulfo Functionalities

  • Protocol
  • First Online:
Glycovirology

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2556))

Abstract

Methods to synthesize influenza virus inhibitors with fluoro, phosphono, and/or sulfo functional groups are described. The resulting sialic acid analogues are produced from the natural substrate N-acetylneuraminic acid as starting material. Fluorescent assay methods for inhibition of influenza neuraminidase and virus proliferation are also provided.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 89.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 119.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Abbreviations

4-MU:

4-Methylumbelliferone

4-MU-Gal:

4-Methylumbelliferyl β-d-galactoside

Ab:

Antibody

anhyd.:

Anhydrous

BSC:

Biosafety cabinet

DAST:

Diethylaminosulfur trifluoride

E:

Enzyme

e.g.:

For example

Em:

Emission wavelength

Ex:

Excitation wavelength

Gal:

Galactose

i.e.:

That is

IAV :

Influenza A virus

IC50:

Half-maximal inhibitory concentration

MDCK:

Madin-Darby canine kidney

MEM:

Minimum essential medium

MES:

2-(N-Morpholino)ethanesulfonic acid

MUNANA:

4-Methylumbelliferyl-Neu5Ac

NA :

Neuraminidase

Neu5Ac :

N-acetylneuraminic acid

NP:

Nucleoprotein

PBS:

Phosphate-buffered salinesat.: Saturated

ST6Gal I:

β-Galactoside α(2–6)-sialyltransferase I soln.: Solution

Tyr:

Tyrosine

References

  1. Muthuri SG, Venkatesan S, Myles PR, Leonardi-Bee J, Al Khuwaitir TS, Al Mamun A, Anovadiya AP et al (2014) Effectiveness of neuraminidase inhibitors in reducing mortality in patients admitted to hospital with influenza A H1N1pdm09 virus infection: a meta-analysis of individual participant data. Lancet Respir Med 2:395–404

    Article  CAS  Google Scholar 

  2. Ishiguro N, Koseki N, Kaiho M, Ariga T, Kikuta H, Oba K et al (2018) Clinical effectiveness of four neuraminidase inhibitors (oseltamivir, zanamivir, laninamivir, and peramivir) for children with influenza A and B in the 2014-2015 to 2016-2017 influenza seasons in Japan. J Infect Chemother 24:449–457

    Article  CAS  Google Scholar 

  3. Chow EJ, Doyle JD, Uyeki TM (2019) Influenza virus-related critical illness: prevention, diagnosis, treatment. Crit Care 23:214

    Article  Google Scholar 

  4. Davidson S (2018) Treating influenza infection, from now and into the future. Front Immunol 9:1946

    Article  Google Scholar 

  5. Lampejo T (2020) Influenza and antiviral resistance: an overview. Eur J Clin Microbiol Infect Dis 39:1201–1208

    Article  Google Scholar 

  6. Lee N, Hurt AC (2018) Neuraminidase inhibitor resistance in influenza: a clinical perspective. Curr Opin Infect Dis 31:520–526

    Article  CAS  Google Scholar 

  7. Sriwilaijaroen N, Vavricka CV, Kiyota H, Suzuki Y (2022) Chapter 21: Influenza A virus neuraminidase inhibitors. In: Suzuki Y (ed) Glycovirology: methods and protocols. Springer, this volume.

    Google Scholar 

  8. Watts AG, Oppezzo P, Withers SG, Alzari PM, Buschiazzo A (2005) Structural and kinetic analysis of two covalent sialosyl-enzyme intermediates on Trypanosoma rangeli sialidase. Structure 281:4149–4155

    Google Scholar 

  9. Vavricka CV, Liu Y, Kiyota H, Sriwilajiaroen N, Qi JX, Tanaka K, Wu Y, Li Q, Li Y, Yan JH, Suzuki Y, Gao GF (2013) Influenza neuraminidase operates via a nucleophilic mechanism and can be targeted by covalent inhibitors. Nat Commun 4:1491. https://doi.org/10.1038/ncomms2487

    Article  CAS  PubMed  Google Scholar 

  10. Li Q, Qi JX, Wu Y, Kiyota H, Tanaka K, Suhara Y, Ohrui H, Suzuki Y, Vavricka CJ, Gao GF (2013) Functional and structural analysis of influenza neuraminidase N3 offers further insight into the mechanisms of oseltamivir resistance. J Virol 87:10016–10024. https://doi.org/10.1128/JVI.01129-13

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Nakajima T, Hori H, Ohrui H, Meguro H, Ido T (1988) Synthesis of N-acetyl-3-fluoro-neuraminic acids. Agric Biol Chem 52:1209–1215

    CAS  Google Scholar 

  12. Hagiwara T, Kijima-Suda I, Ido T, Ohrui H, Tomita K (1994) Inhibition of bacterial and viral silaidases by 3-fluoro-N-acetylneuraminic acid. Carbohydr Res 263:167–172

    Article  CAS  Google Scholar 

  13. Wallimann K, Vasella A (1990) Phosphonic-acid analogues of the N-Acetyl-2-deoxyneuraminic acids: synthesis and inhibition of Vibrio cholerae sialidase. Helv Chim Acta 73:1359–1372

    Article  CAS  Google Scholar 

  14. Vasella A, Wyler R (1991) Synthesis of a phosphonic acid analogue of N-acetyl-2,3-didehydro-2-deoxyneuraminic acid, an inhibitor of Vibrio cholerae sialidase. Helv Chim Acta 1991(74):451–463

    Article  Google Scholar 

  15. Vavricka CJ, Muto C, Hasunuma T, Kimura Y, Araki M, Wu Y, Gao GF, Ohrui H, Izumi M, Kiyota H (2017) Synthesis of sulfo-sialic acid analogues: potent neuraminidase inhibitors in regards to anomeric functionality. Sci Rep 7:8239

    Article  Google Scholar 

  16. Watts AG, Withers SG (2004) The synthesis of some mechanistic probes for sialic acid processing enzymes and the labeling of a sialidase from Trypanosoma rangeli. Can J Chem 82:1581–1588

    Article  CAS  Google Scholar 

  17. Morais GR, Oliveira RS, Falconer RA (2009) Selective synthesis of Neu5Ac2en and its oxazoline derivative using BF3·Et2O. Tetrahedron Lett 50:1642–1644

    Article  CAS  Google Scholar 

  18. Burkart MD, Zhiyuan Zhang Z, Hung SC, Wong CH (1997) A new method for the synthesis of fluoro-carbohydrates and glycosides using selectfluor. J Am Chem Soc 119:11743–11746

    Article  CAS  Google Scholar 

  19. Shie JJ, Fang JM, Lai PT, Wen WH, Wang SY, Cheng YSE, Tsai KC, Yang AS, Wong CH (2011) A practical synthesis of zanamivir phosphonate congeners with potent anti-influenza activity. J Am Chem Soc 133:17959–17965

    Article  CAS  Google Scholar 

  20. Potier M, Mameli L, Belisle M, Dallaire L, Melancon SB (1979) Fluorometric assay of neuraminidase with a sodium (4-methylumbelliferyl-α-d-N-acetylneuraminate) substrate. Anal Biochem 94:287–296

    Article  CAS  Google Scholar 

  21. Nakakita SI, Sriwilaijaroen N, Suzuki Y et al (2020) Preparation and detection of glycan-binding activity of influenza virus. Methods Mol Biol 2132:567–583

    Article  CAS  Google Scholar 

  22. Sriwilaijaroen N, Kondo S, Yagi H et al (2009) Analysis of N-glycans in embryonated chicken egg chorioallantoic and amniotic cells responsible for binding and adaptation of human and avian influenza viruses. Glycoconj J 26:433–443

    Article  CAS  Google Scholar 

  23. Sriwilaijaroen N, Nakakita SI, Kondo S et al (2018) N-glycan structures of human alveoli provide insight into influenza A virus infection and pathogenesis. FEBS J 285:1611–1634

    Article  CAS  Google Scholar 

  24. Hatakeyama S, Sakai-Tagawa Y, Kiso M et al (2005) Enhanced expression of an α2-6-linked sialic acid on MDCK cells improves isolation of human influenza viruses and evaluation of their sensitivity to a neuraminidase inhibitor. J Clin Microbiol 43:4139–4146

    Article  CAS  Google Scholar 

  25. Sriwilaijaroen N, Kadowaki A, Onishi Y et al (2011) Mumefural and related HMF derivatives from Japanese apricot fruit juice concentrate show multiple inhibitory effects on pandemic influenza A (H1N1) virus. Food Chem 127:1–9

    Article  CAS  Google Scholar 

  26. Sriwilaijaroen N, Suzuki K, Takashita E et al (2015) 6SLN-lipo PGA specifically catches (coats) human influenza virus and synergizes neuraminidase-targeting drugs for human influenza therapeutic potential. J Antimicrob Chemother 70:2797–2809

    Article  CAS  Google Scholar 

  27. Sriwilaijaroen N, Fukumoto S, Kumagai K et al (2012) Antiviral effects of Psidium guajava Linn. (guava) tea on the growth of clinical isolated H1N1 viruses: its role in viral hemagglutination and neuraminidase inhibition. Antivir Res 94:139–146

    Article  CAS  Google Scholar 

  28. Sriwilaijaroen N, Magesh S, Imamura A et al (2016) A novel potent and highly specific inhibitor against influenza viral N1-N9 neuraminidases: insight into neuraminidase-inhibitor interactions. J Med Chem 59:4563–4577

    Article  CAS  Google Scholar 

  29. Takahashi T, Murakami K, Nagakura M et al (2008) Sulfatide is required for efficient replication of influenza A virus. J Virol 82:5940–5950

    Article  CAS  Google Scholar 

  30. Klimov A, Balish A, Veguilla V et al (2012) Influenza virus titration, antigenic characterization, and serological methods for antibody detection. Methods Mol Biol 865:25–25

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Graduate School of Science, Technology and Innovation, Kobe University, Kobe, Japan

    Christopher J. Vavricka

  2. Department of Preclinical Sciences, Faculty of Medicine, Thammasat University, Pathumthani, Thailand

    Nongluk Sriwilaijaroen

  3. Department of Medical Biochemistry, School of Pharmaceutical Sciences, University of Shizuoka, Shizuoka, Japan

    Nongluk Sriwilaijaroen & Yasuo Suzuki

  4. Graduate School of Environmental and Life Science, Okayama University, Okayama, Japan

    Hiromasa Kiyota

Authors
  1. Christopher J. Vavricka
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Nongluk Sriwilaijaroen
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yasuo Suzuki
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Hiromasa Kiyota
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Christopher J. Vavricka or Hiromasa Kiyota .

Editor information

Editors and Affiliations

  1. Department of Medical Biochemistry, University of Shizuoka School of Pharmaceutical Sciences, Shizuoka, Japan

    Yasuo Suzuki

Rights and permissions

Reprints and permissions

Copyright information

© 2022 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Vavricka, C.J., Sriwilaijaroen, N., Suzuki, Y., Kiyota, H. (2022). Synthesis and Neuraminidase Inhibitory Activity of Sialic Acid Analogues with Fluoro, Phosphono, and Sulfo Functionalities. In: Suzuki, Y. (eds) Glycovirology. Methods in Molecular Biology, vol 2556. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2635-1_20

Download citation

  • DOI: https://doi.org/10.1007/978-1-0716-2635-1_20

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2634-4

  • Online ISBN: 978-1-0716-2635-1

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation