Differences in binding kinetics, bond strength and adduct formation between Pt-based drugs and S- or N-donor groups: A comparative study using mass spectrometry techniques

Pt-S and Pt-N interactions resulting from the coordination of cisplatin, oxaliplatin and carboplatin to two synthetic peptides that differ from each other in one amino acid (Met or His) have been thoroughly studied in this work. The degree of Pt-binding was determined by inductively coupled plasma mass spectrometry after the separation of the Pt-complexes from the unreacted drugs by size exclusion chromatography. Cisplatin and oxaliplatin showed high affinity for the peptides from the first hours of incubation, although the peptides required longer incubation times to obtain the same platination degrees with cisplatin than with oxaliplatin. Once the reactions reached their maximum binding degrees, the complexes with oxaliplatin began to dissociate, revealing binding reversibility, while a pseudo steady-state was observed for cisplatin until the last day of incubation. Conversely, the equilibrium was not reached for carboplatin and the His-peptide after 30 days, showing a binding degree of 16%, versus 78% for the Met-peptide. The S-donor group also presented an important influence on the reactivity and the adduct formation. The reaction rate for the Mat-peptide was faster than the hydrolysis of oxaliplatin and carboplatin, and all the drugs, except oxaliplatin, were able to coordinate up to 3 different donor groups, which were identified by nanospray mass spectrometry. Since structural characterization of metal-complexes often represents an analytical challenge during electrophoretic separations, the strength of Pt-Met and Pt-His bonds was also evaluated under these conditions. The nature of the electrophoretic agents and the incubation times used were the parameters that most affected the stability. Higher Pt losses were found for the Met-peptide (35-90%) than for the His-peptide (16-48%), indicating that Pt-Met bonds were kinetically preferred while Pt-His interactions were thermodynamically favored.