Positive effect of natural and negatively charged cyclodextrins on the stabilization of penicillins towards β-lactamase degradation due to inclusion and external guest–host association. An NMR and MS study

Abstract

The complexation of penicillin (1a–c) and cephalosporin (2a,b) antibiotics with cyclodextrins (CDs), both natural [β-CD (3b) and γ-CD (3c)] and carboxylated [heptakis(6-oxycarbonylethylthio-6-deoxy)-β-CD sodium salt (4b) and octakis(6-oxycarbonylethylthio-6-deoxy)-γ-CD (4c) sodium salt], has been studied at neutral pH. Penicillins [ampicillin (1a), amoxicillin (1b) and dicloxacillin (1c) form inclusion complexes with the above CDs, as was shown by extensive NMR spectroscopic studies, whereas cephalosporins (cephalexin, cefadroxil) do not. Inclusion of the penicillins into either 3b or 4b was not accompanied by significant chemical shift changes in the ¹H NMR spectra. On the contrary, with the wider 3c and its derivative 4c inclusion was evidenced by both chemical shift displacements of the cavity protons and intermolecular interactions, indicating the formation of primarily 1 : 1 guest–host inclusion complexes. The binding constants for 1a/3c, 1a/4c and 1c/3c were calculated as 19 ± 4, 17 ± 0.9 and 622 ± 200 M⁻¹, respectively. With 4c, a 1 : 2 stoichiometry was also found. In addition, simultaneous formation of aggregates by external association takes place in solution, as shown by the ESI-mass spectrometric data. Studies on the hydrolysis of ampicillin under pseudo-first order conditions using an excess of 3c, 4c and of linear maltoheptaose at pH 7 showed that the drug hydrolysed at a similar rate in all cases. In the presence, however, of β-lactamase enzyme and the carboxylated host 4c, ampicillin degraded twice as slowly (0.008 h⁻¹) as in the presence of β-lactamase alone (0.017 h⁻¹). This was explained by the effective protection provided by both inclusion and external association of the host. The interaction, therefore, of penicillins with carboxylated CDs may present a means to lessen the chemical instability of these drugs in the presence of β-lactamase enzymes.