title: A strategy for neuraminidase inhibitors using mechanism-based labeling information.
abstract: A potent inhibitor for Vibrio cholerae neuraminidase (VCNA) was developed by using a novel two-step strategy, a target amino acid validation using mechanism-based labeling information, and a potent inhibitor search using a focused library. The labeling information suggested the hidden dynamics of a loop structure of VCNA, which can be a potential target of the novel inhibitor. A focused library composed of 187 compounds was prepared from a 9-azide derivative of 2,3-dehydro-N-acetylneuraminic acid (DANA) to interrupt the function of the loop of the labeled residues. Inhibitor 3 c showed potent inhibition properties and was the strongest inhibitor with FANA, a N-trifluoroacetyl derivative of DANA. Validation studies of the inhibitor with a detergent and a Lineweaver-Burk plot suggested that the 9-substitution group would interact hydrophobically with the target loop moiety, adding a noncompetitive inhibition property to the DANA skeleton. This information enabled us to design compound 4 having the combined structure of 3 c and FANA. Compound 4 showed the most potent inhibition (K(i) =73 nM, mixed inhibition) of VCNA with high selectivity among the tested viral, bacterial, and mammal neuraminidases.
authors.affiliation: Graduate School of Life Science and Frontier Research, Center for the Post-Genome Science and Technology, Hokkaido University, Kita-ku, Sapporo, Japan. hinou@glyco.sci.hokudai.ac.jp
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Hinou
authors.forename: Risho
authors.initials: R
authors.lastname: Miyoshi
authors.forename: Yasuaki
authors.initials: Y
authors.lastname: Takasu
authors.forename: Hirokazu
authors.initials: H
authors.lastname: Kai
authors.forename: Masaki
authors.initials: M
authors.lastname: Kurogochi
authors.forename: Shingo
authors.initials: S
authors.lastname: Arioka
authors.forename: Xiao-Dong
authors.initials: XD
authors.lastname: Gao
authors.forename: Nobuaki
authors.initials: N
authors.lastname: Miura
authors.forename: Naoki
authors.initials: N
authors.lastname: Fujitani
authors.forename: Shinya
authors.initials: S
authors.lastname: Omoto
authors.forename: Tomokazu
authors.initials: T
authors.lastname: Yoshinaga
authors.forename: Tamio
authors.initials: T
authors.lastname: Fujiwara
authors.forename: Takeshi
authors.initials: T
authors.lastname: Noshi
authors.forename: Hiroko
authors.initials: H
authors.lastname: Togame
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Takemoto
authors.forename: Shin-Ichiro
authors.initials: S
authors.lastname: Nishimura
mesh_terms: D020134:Catalytic Domain; D015195:Drug Design; D004791:Enzyme Inhibitors / Q000737:chemistry* / Q000494:pharmacology; D009439:Neuraminidase / Q000037:antagonists & inhibitors* / Q000378:metabolism; D017434:Protein Structure, Tertiary; D012794:Sialic Acids / Q000737:chemistry; D013329:Structure-Activity Relationship; D014734:Vibrio cholerae / Q000201:enzymology
chemical_list: D004791:Enzyme Inhibitors; D012794:Sialic Acids; D009439:Neuraminidase