La multi-ligation triazole : Développement de nouveaux outils pour la synthèse de mimes de protéines par cycloadditions successives

Abstract : The aim of this work was the development of a novel method for the synthesis of triazolo-proteins by multiple successive copper-catalyzed azide-alkyne cycloadditions (CuAAC). In order to achieve several successive cycloadditions, we have studied the stability and cleavage conditions of several alkyne protective groups. This study led us to the development of an original strategy in order to achieve three successive cycloadditions on a same scaffold by temporal protection of alkyne functionalities. The method has been applied to the synthesis of an analogue of human stefin A, a natural inhibitor of several therapeutically relevant cysteine proteases. Therefore, we have developed CuAAC conditions compatible with unprotected peptide ligation. The strategy allowed us to obtain a bis-triazolo analogue of human stefin A. Circular dichroism and enzymology assays on several cysteine cathepsins revealed that the synthetic analogue has retained the folding and full biological activity of the native protein. In order to expand the possibilities of this strategy, we have developed reaction conditions allowing us to perform successive triazole ligation on solid phase. This methodology avoids the need for a time-consuming and labor-intensive purification step before and after each ligation. With the aim of exploring the use of analogues of the tumor-associated form of the glycoprotein MUC1 to induce a specific immune response, we have synthesized a triazolo-analogue of MUC1 of 160 aminoacids using solid phase peptide ligation.
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Ibai Valverde. La multi-ligation triazole : Développement de nouveaux outils pour la synthèse de mimes de protéines par cycloadditions successives. Chimie. Universite d'Orléans, 2010. Français. ⟨tel-01157275⟩

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