An Efficient Approach to the Discovery of Potent Inhibitors against Glycosyltransferases

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• 作者：Kensaku Hosoguchi ; Takahiro Maeda ; Jun-ichi Furukawa ; Yasuro Shinohara ; Hiroshi Hinou ; Mitsuaki Sekiguchi ; Hiroko Togame ; Hiroshi Takemoto ; Hirosato Kondo ; Shin-Ichiro Nishimura
• 刊名：Journal of Medicinal Chemistry
• 出版年：2010
• 出版时间：August 12, 2010
• 年：2010
• 卷：53
• 期：15
• 页码：5607-5619
• 全文大小：1291K
• 年卷期：v.53,no.15(August 12, 2010)
• ISSN：1520-4804

文摘

We describe a standardized approach for searching potent and selective inhibitors of glycosyltransferases by high throughput quantitative MALDI-TOFMS-based screening of focused compound libraries constructed by 1,3-dipolar cycloaddition of the desired azidosugar nucleotides with various alkynes. An aminooxy-functionalized reagent with a stable isotope was conjugated with oligosaccharides to afford glycopeptides as acceptor substrates with improved ion sensitivity. Enhanced ionization potency of new substrates allowed for MALDI-TOFMS-based facile and quantitative analysis of enzymatic glycosylation in the presence of glycosyl donor substrates. A non-natural synthetic sugar nucleotide was identified to be the first highly specific inhibitor for rat recombinant α2,3-(N)-sialyltransferase (α2,3ST, IC₅₀ = 8.2 μM), while this compound was proved to become a favorable substrate for rat recombinant α2,6-(N)-sialyltransferase (α2,6ST, K_m = 125 μM). Versatility of this strategy was demonstrated by identification of two selective inhibitors for human recombinant α1,3-fucosyltransferase V (α1,3-FucT, K_i = 293 nM) and α1,6-fucosyltransferase VIII (α1,6-FucT, K_i = 13.8 μM).