An Efficient Approach to the Discovery of Potent Inhibitors against Glycosyltransferases

作者：Hosoguchi Kensaku; Maeda Takahiro; Furukawa Jun ichi; Shinohara Yasuro; Hinou Hiroshi; Sekiguchi Mitsuaki; Togame Hiroko; Takemoto Hiroshi; Kondo Hirosato; Nishimura Shin Ichiro^*

来源：Journal of Medicinal Chemistry, 2010, 53(15): 5607-5619.

DOI：10.1021/jm100612r

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 alpha 2,3-(N)-sialyltransferase (alpha 2,3ST, IC(50) = 8.2 mu M), while this compound was proved to become a favorable substrate for rat recombinant alpha 2,6-(N)-sialyltransferase (alpha 2,6ST, K(m) = 125 mu M). Versatility of this strategy was demonstrated by identification of two selective inhibitors for human recombinant alpha 1,3-fucosyltransferase V (alpha 1,3-FucT, K(i) = 293 nM) and alpha 1,6-fucosyltransferase VIII (alpha 1,6-FucT, K(i) = 13.8 mu M).

• 出版日期2010-8-12