A nanomolar multivalent ligand as entry inhibitor of the hemagglutinin of avian influenza.

Influenza virus attaches itself to sialic acids on the surface of epithelial cells of the upper respiratory tract of the host using its own protein hemagglutinin. Species specificity of influenza virus is determined by the linkages of the sialic acids. Birds and humans have α2-3 and α2-6 linked sialic acids, respectively. Viral hemagglutinin is a homotrimeric receptor, and thus, tri- or oligovalent ligands should have a high binding affinity. We describe the in silico design, chemical synthesis and binding analysis of a trivalent glycopeptide mimetic. This compound binds to hemagglutinin H5 of avian influenza with a dissociation constant of K(D) = 446 nM and an inhibitory constant of K(I) = 15 μM. In silico modeling shows that the ligand should also bind to hemagglutinin H7 of the virus that causes the current influenza outbreak in China. The trivalent glycopeptide mimetic and analogues have the potential to block many different influenza viruses.

[1]  Xiangqun Zeng,et al.  Engineering peptide linkers for scFv immunosensors. , 2008, Analytical chemistry.

[2]  David J. Stevens,et al.  Haemagglutinin mutations responsible for the binding of H5N1 influenza A viruses to human-type receptors , 2006, Nature.

[3]  G. Air,et al.  Structure of influenza virus neuraminidase B/Lee/40 complexed with sialic acid and a dehydro analog at 1.8-A resolution: implications for the catalytic mechanism. , 1994, Biochemistry.

[4]  H. Klenk,et al.  Neuraminidase Is Important for the Initiation of Influenza Virus Infection in Human Airway Epithelium , 2004, Journal of Virology.

[5]  I D Kuntz,et al.  Inhibition of the fusion-inducing conformational change of influenza hemagglutinin by benzoquinones and hydroquinones. , 1993, Biochemistry.

[6]  A. Gamian,et al.  Inhibition of influenza A virus hemagglutinin and induction of interferon by synthetic sialylated glycoconjugates. , 1991, Canadian journal of microbiology.

[7]  Ian A. Wilson,et al.  Structure and Receptor Specificity of the Hemagglutinin from an H5N1 Influenza Virus , 2006, Science.

[8]  H. Yen,et al.  Targeting the host or the virus: Current and novel concepts for antiviral approaches against influenza virus infection , 2012, Antiviral Research.

[9]  Hideo Goto,et al.  Avian flu: Isolation of drug-resistant H5N1 virus , 2005, Nature.

[10]  Y. Cheng,et al.  Relationship between the inhibition constant (K1) and the concentration of inhibitor which causes 50 per cent inhibition (I50) of an enzymatic reaction. , 1973, Biochemical pharmacology.

[11]  Jason E Gestwicki,et al.  Influencing receptor-ligand binding mechanisms with multivalent ligand architecture. , 2002, Journal of the American Chemical Society.

[12]  J. Knowles,et al.  Design and evaluation of a tightly binding fluorescent ligand for influenza A hemagglutinin , 1992 .

[13]  Thomas Peters,et al.  NMR spectroscopy techniques for screening and identifying ligand binding to protein receptors. , 2003, Angewandte Chemie.

[14]  J. Skehel,et al.  X-ray structures of H5 avian and H9 swine influenza virus hemagglutinins bound to avian and human receptor analogs , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[15]  P. Kitov,et al.  On the nature of the multivalency effect: a thermodynamic model. , 2003, Journal of the American Chemical Society.

[16]  P. Lutz,et al.  Synthese anomerer Sialinsäure‐methylketoside , 1966 .

[17]  G. Whitesides,et al.  Polyacrylamides Bearing Pendant α-Sialoside Groups Strongly Inhibit Agglutination of Erythrocytes by Influenza Virus: The Strong Inhibition Reflects Enhanced Binding through Cooperative Polyvalent Interactions , 1996 .

[18]  R. Webster,et al.  The Influenza Virus Enigma , 2009, Cell.

[19]  C. Boucher,et al.  Emergence of a multidrug-resistant pandemic influenza A (H1N1) virus. , 2010, The New England journal of medicine.

[20]  R. Webster,et al.  Are We Ready for Pandemic Influenza? , 2003, Science.

[21]  Wei Zhang,et al.  Discovery of the first series of small molecule H5N1 entry inhibitors. , 2009, Journal of medicinal chemistry.

[22]  Hua Yang,et al.  Structures of Receptor Complexes of a North American H7N2 Influenza Hemagglutinin with a Loop Deletion in the Receptor Binding Site , 2010, PLoS pathogens.

[23]  Mark von Itzstein,et al.  The war against influenza: discovery and development of sialidase inhibitors , 2007, Nature Reviews Drug Discovery.

[24]  David J Stevens,et al.  Structure of influenza hemagglutinin in complex with an inhibitor of membrane fusion , 2008, Proceedings of the National Academy of Sciences.

[25]  G M Whitesides,et al.  A trivalent system from vancomycin.D-ala-D-Ala with higher affinity than avidin.biotin. , 1998, Science.

[26]  I. Wilson,et al.  Structure of the haemagglutinin membrane glycoprotein of influenza virus at 3 Å resolution , 1981, Nature.

[27]  George M Whitesides,et al.  Polyvalent Interactions in Biological Systems: Implications for Design and Use of Multivalent Ligands and Inhibitors. , 1998, Angewandte Chemie.

[28]  Shuwen Liu,et al.  Influenza A Virus Entry Inhibitors Targeting the Hemagglutinin , 2013, Viruses.

[29]  T. Sakai,et al.  Roles of neuraminidase in the initial stage of influenza virus infection. , 2006, Microbes and infection.

[30]  H. Ogura,et al.  Studies on sialic acids. V: Synthesis of α- and β-D-Neu5Acp-(2→6)-lactose , 1986 .

[31]  J. Knowles,et al.  Molecular recognition of bivalent sialosides by influenza virus , 1991 .

[32]  G M Whitesides,et al.  Hemagglutinins from two influenza virus variants bind to sialic acid derivatives with millimolar dissociation constants: a 500-MHz proton nuclear magnetic resonance study. , 1989, Biochemistry.

[33]  Bernd Meyer,et al.  Characterization of Ligand Binding by Saturation Transfer Difference NMR Spectroscopy. , 1999, Angewandte Chemie.

[34]  R. Read,et al.  Shiga-like toxins are neutralized by tailored multivalent carbohydrate ligands , 2000, Nature.

[35]  J. Knowles,et al.  Monovalent sialosides that bind tightly to influenza A virus. , 1991, Journal of medicinal chemistry.