Synthesis of Glycoconjugate Vaccines for Candida albicans Using Novel Linker Methodology

The cell wall phosphomannan of Candida species is a complex N-linked glycoprotein with a glycan chain that contains predominantly α-linked mannose residues. However, it is the minor β-mannan component of the phosphomannan of clinically important Candida strains that provides immunological protection in animal models of fungal disease and hence holds promise as a component of conjugate vaccines. This important antigen occurs in different forms linked to the α-mannan backbone via a phosphodiester bond (acid-labile β-mannan) or directly via a glycosidic bond. To reproducibly synthesize and evaluate conjugate vaccines, a robust method for the synthesis of the different oligosaccharide epitopes is required. Here, we report the gram-scale syntheses of both types of epitopes by an approach that utilizes glucosyl trichloroacetimidate donor 2 to first create a β-glucopyranoside linkage and then epimerizes the C-2 center via an oxidation−reduction sequence that provides an efficient multigram scale route to the β-mannopyranosides 5, 8, and 15. Reaction of glycosides 16−18 with homobifunctional adipic acid p-nitrophenyl diesters in dry DMF gave the corresponding half esters in good yields, and of sufficient stability to permit chromatographic purification. Subsequent conjugation with BSA and tetanus toxiod (TT) under mild conjugation conditions afforded the corresponding tri- and tetrasaccharide neoglycoproteins with good efficiency. The conjugation method is also applicable to the coupling of small amounts (mg) of larger oligosaccharides with different proteins.