Synthesis of Le aLe x oligosaccharide fragments and efficient one-step deprotection

Abstract

We describe here the synthesis of two oligosaccharide fragments of the tumor associated carbohydrate antigen Le aLe x. While the linear lacto- N-triose I: β- d-Gal p-(1→4)-β- d-GlcNAc p-(1→3)-β- d-Gal p-OMe is a known compound, this is the first reported preparation of the branched tetrasaccharide β- d-GlcNAc p-(1→3)-β- d-Gal p-(1→4)-[α- l-Fuc p-(1→3)]-β- d-GlcNAc p-OMe. Our synthetic schemes involved using an N-trichloroacetylated trichloroacetimidate glucosaminyl donor activated with excess TMSOTf at 0 °C for glycosylation at O-3 of galactosyl residues and that of trichloroacetimidate galactosyl donors activated with excess BF 3·OEt 2 to glycosylate either O-3 or O-4 of glucosamine residues. The fucosylation at O-3 of the glucosamine acceptor was accomplished using a thiofucoside donor activated with copper(II) bromide and tetrabutylammonium bromide. Thus, syntheses of the protected tri- and tetrasaccharides were achieved easily and efficiently using known building blocks. Of particular interest, we also report that these protected oligosaccharides were submitted to dissolving metal conditions (Na–NH 3) to provide in one single step the corresponding deprotected compounds. Under these conditions all protecting groups ( O-acyl, benzylidene, benzyl, and N-trichloroacetyl) were efficiently cleaved. The work-up procedure for such reactions usually involves quenching with excess methanol and then neutralization with acetic acid. In our work the neutralization was carried out using acetic anhydride rather than acetic acid to ensure N-acetylation of the glucosamine residue. Both fully deprotected compounds were then simply purified and desalted by gel permeation chromatography on a Biogel P2 column eluted with water.