Molecular Recognition of Sialic Acid by Lanthanide(III) Complexes through Cooperative Two-Site Binding

Abstract

Herein we report two new ligands, 1,4,7-tris(carboxymethyl)-10-[2-(dihydroxyboranyl)benzyl]-1,4,7,10-tetraazacyclododecane (L(1)) and 1,4,7-tris(carboxymethyl)-10-[3-(dihydroxyboranyl)benzyl]-1,4,7,10-tetraazacyclododecane (L(2)), which contain a phenylboronic acid (PBA) function and a 1,4,7,10-tetraazacyclododecane-1,4,7-triacetate cage for complexation of lanthanide ions in an aqueous solution. The pK(a) of the PBA function amounts to 4.6 in [Gd(L(1))] and 8.9 in [Gd(L(2))], with the value of the L(2) analogue being very similar to that of PBA (8.8). These results are explained by the coordination of the PBA function of L(1) to the Gd(III) ion, which results in a dramatic lowering of its pK(a). As a consequence, [Gd(L(1))] does not bind to saccharides at physiological pH. The nuclear magnetic relaxation dispersion profiles recorded for [Gd(L(1))] and [Gd(L(2))] confirm that the phenylboronate function is coordinated to the metal ion in the L(1) derivative, which results in a q = 0 complex. The interaction of the [Gd(L(2))] complex with 5-acetylneuraminic acid (Neu5Ac) and 2-alpha-O-methyl-5-acetylneuraminic acid (MeNeu5Ac) has been investigated by means of spectrophotometric titrations in an aqueous solution (pH 7.4, 0.1 M 3-(N-morpholino)propanesulfonic acid buffer). Furthermore, we have also investigated the binding of these receptors with competing monosaccharides such as D-(+)-glucose, D-fructose, D-mannose, D-galactose, methyl alpha-D-galactoside, and methyl alpha-D-mannoside. The binding constants obtained indicate an important selectivity of [Gd(L(2))] for Neu5Ac (K(eq) = 151) over D-(+)-glucose (K(eq) = 12.3), D-mannose (K(eq) = 21.9), and D-galactose (K(eq) = 24.5). Furthermore, a very weak binding affinity was observed in the case of methyl alpha-D-galactoside and methyl alpha-D-mannoside. An 8-fold increase of the binding constant of [Gd(L(2))] with Neu5Ac is observed when compared to that of PBA determined under the same conditions (K(eq) = 19). (13)C NMR spectroscopy and density functional theory calculations performed at the B3LYP/6-31G(d) level show that this is due to a cooperative two-site binding of Neu5Ac through (1) ester formation by interaction on the PBA function of the receptor and (2) coordination of the carboxylate group of Neu5Ac to the Gd(III) ion. The emission lifetime of the (5)D(4) level of Tb(III) in [Tb(L(2))] increases upon Neu5Ac binding, in line with the displacement of inner-sphere water molecules due to coordination of Neu5Ac to the metal ion.