title: Structure-Based Tetravalent Zanamivir with Potent Inhibitory Activity against Drug-Resistant Influenza Viruses.
abstract: Zanamivir and oseltamivir are principal influenza antiviral drugs that target viral neuraminidase (NA), but resistant viruses containing mutant NAs with diminished drug affinity are increasingly emerging. Using the structural knowledge of both drug-binding sites and their spatial arrangement on the homotetrameric NA, we have developed a tetravalent zanamivir (TZ) molecule that exhibited marked increases in NA binding affinity, inhibition of NA enzyme activity, and in vitro plus in vivo antiviral efficacy over zanamivir. TZ functioned against both human seasonal H3N2 and avian H7N9 viruses, including drug-resistant mutants. Crystal structure of a resistant N9 NA in complex with TZ explained the function, which showed that four zanamivir residues simultaneously bound to all four monomers of NA. The design method of TZ described in this study may be useful to develop drugs or ligands that target proteins with multiple binding sites. The potent anti-influenza activity of TZ makes it attractive for further development.
authors.affiliation: CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences , Chaoyang District, Beijing 100101, China.|National Engineering Research Center for Carbohydrate Synthesis, Jiangxi Normal University , Nanchang 330022, China.|Graduate University of Chinese Academy of Sciences , Shijingshan District, Beijing 100049, China.
authors.forename: Lifeng
authors.initials: L
authors.lastname: Fu
authors.affiliation: CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences , Chaoyang District, Beijing 100101, China.
authors.forename: Yuhai
authors.initials: Y
authors.lastname: Bi
authors.affiliation: CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences , Chaoyang District, Beijing 100101, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Wu
authors.affiliation: CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences , Chaoyang District, Beijing 100101, China.|Graduate University of Chinese Academy of Sciences , Shijingshan District, Beijing 100049, China.
authors.forename: Shanshan
authors.initials: S
authors.lastname: Zhang
authors.affiliation: CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences , Chaoyang District, Beijing 100101, China.
authors.forename: Jianxun
authors.initials: J
authors.lastname: Qi
authors.affiliation: CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences , Chaoyang District, Beijing 100101, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Li
authors.affiliation: Laboratory Animal Center, Chinese Center for Disease Control and Prevention , Changping District, Beijing 102206, China.
authors.forename: Xuancheng
authors.initials: X
authors.lastname: Lu
authors.affiliation: CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences , Chaoyang District, Beijing 100101, China.|Graduate University of Chinese Academy of Sciences , Shijingshan District, Beijing 100049, China.
authors.forename: Zhenning
authors.initials: Z
authors.lastname: Zhang
authors.affiliation: CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences , Chaoyang District, Beijing 100101, China.
authors.forename: Xun
authors.initials: X
authors.lastname: Lv
authors.affiliation: CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences , Chaoyang District, Beijing 100101, China.|Center for Influenza Research and Early Warning, Chinese Academy of Sciences (CASCIRE) , Chaoyang District, Beijing 100101, China.
authors.forename: Jinghua
authors.initials: J
authors.lastname: Yan
authors.affiliation: CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences , Chaoyang District, Beijing 100101, China.|Center for Influenza Research and Early Warning, Chinese Academy of Sciences (CASCIRE) , Chaoyang District, Beijing 100101, China.
authors.forename: George F
authors.initials: GF
authors.lastname: Gao
authors.affiliation: CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences , Chaoyang District, Beijing 100101, China.|National Engineering Research Center for Carbohydrate Synthesis, Jiangxi Normal University , Nanchang 330022, China.|Center for Influenza Research and Early Warning, Chinese Academy of Sciences (CASCIRE) , Chaoyang District, Beijing 100101, China.
authors.forename: Xuebing
authors.initials: X
authors.lastname: Li
mesh_terms: D000818:Animals; D000998:Antiviral Agents / Q000737:chemistry / Q000494:pharmacology*; D018360:Crystallography, X-Ray; D004285:Dogs; D004305:Dose-Response Relationship, Drug; D024882:Drug Resistance, Viral / Q000187:drug effects*; D006801:Humans; D053122:Influenza A Virus, H3N2 Subtype / Q000187:drug effects*; D064766:Influenza A Virus, H7N9 Subtype / Q000187:drug effects*; D061985:Madin Darby Canine Kidney Cells / Q000187:drug effects / Q000821:virology; D051379:Mice; D008807:Mice, Inbred BALB C; D008826:Microbial Sensitivity Tests; D008958:Models, Molecular; D015394:Molecular Structure; D051381:Rats; D017207:Rats, Sprague-Dawley; D013329:Structure-Activity Relationship; D053243:Zanamivir / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*
chemical_list: D000998:Antiviral Agents; D053243:Zanamivir