title: Influenza neuraminidase operates via a nucleophilic mechanism and can be targeted by covalent inhibitors.
abstract: Development of novel influenza neuraminidase inhibitors is critical for preparedness against influenza outbreaks. Knowledge of the neuraminidase enzymatic mechanism and transition-state analogue, 2-deoxy-2,3-didehydro-N-acetylneuraminic acid, contributed to the development of the first generation anti-neuraminidase drugs, zanamivir and oseltamivir. However, lack of evidence regarding influenza neuraminidase key catalytic residues has limited strategies for novel neuraminidase inhibitor design. Here, we confirm that influenza neuraminidase conserved Tyr406 is the key catalytic residue that may function as a nucleophile; thus, mechanism-based covalent inhibition of influenza neuraminidase was conceived. Crystallographic studies reveal that 2α,3ax-difluoro-N-acetylneuraminic acid forms a covalent bond with influenza neuraminidase Tyr406 and the compound was found to possess potent anti-influenza activity against both influenza A and B viruses. Our results address many unanswered questions about the influenza neuraminidase catalytic mechanism and demonstrate that covalent inhibition of influenza neuraminidase is a promising and novel strategy for the development of next-generation influenza drugs.
authors: Research Network of Immunity and Health, Beijing Institutes of Life Science, Beijing 100101, China.
authors: Christopher J
authors: CJ
authors: Vavricka
authors: Yue
authors: Y
authors: Liu
authors: Hiromasa
authors: H
authors: Kiyota
authors: Nongluk
authors: N
authors: Sriwilaijaroen
authors: Jianxun
authors: J
authors: Qi
authors: Kosuke
authors: K
authors: Tanaka
authors: Yan
authors: Y
authors: Wu
authors: Qing
authors: Q
authors: Li
authors: Yan
authors: Y
authors: Li
authors: Jinghua
authors: J
authors: Yan
authors: Yasuo
authors: Y
authors: Suzuki
authors: George F
authors: GF
authors: Gao
mesh_terms: D000818:Animals; D000998:Antiviral Agents / Q000737:chemistry / Q000494:pharmacology*; D055162:Biocatalysis / Q000187:drug effects; D017124:Conserved Sequence; D018360:Crystallography, X-Ray; D024882:Drug Resistance, Viral / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000494:pharmacology*; D005453:Fluorescence; D006801:Humans; D007700:Kinetics; D061985:Madin Darby Canine Kidney Cells; D009682:Magnetic Resonance Spectroscopy; D008958:Models, Molecular; D050505:Mutant Proteins / Q000378:metabolism; D019158:N-Acetylneuraminic Acid / Q000737:chemistry; D009439:Neuraminidase / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000378:metabolism*; D009975:Orthomyxoviridae / Q000187:drug effects / Q000201:enzymology* / Q000502:physiology; D053139:Oseltamivir / Q000494:pharmacology; D014443:Tyrosine / Q000378:metabolism; D014779:Virus Replication / Q000187:drug effects; D053243:Zanamivir / Q000494:pharmacology
chemical_list: D000998:Antiviral Agents; D004791:Enzyme Inhibitors; D050505:Mutant Proteins; D053139:Oseltamivir; D014443:Tyrosine; D009439:Neuraminidase; D019158:N-Acetylneuraminic Acid; D053243:Zanamivir