Journal Article

Biological activity of Pt IV prodrugs triggered by riboflavin-mediated bioorthogonal photocatalysis

Terenzi, Alessio, Martínez, Javier Calvo, Galanski, Markus, Keppler, Bernhard K, Faraone, Adriana, Berger, Walter, Hager, Sonja, Englinger, Bernhard, Alonso-de Castro, Silvia, Salassa, Luca

2018

Overview

We have recently demonstrated that riboflavin (Rf) functions as unconventional bioorthogonal photocatalyst for the activation of Pt prodrugs. In this study, we show how the combination of light and Rf with two Pt prodrugs is a feasible strategy for light-mediated pancreatic cancer cell death induction. In Capan-1 cells, which have high tolerance against photodynamic therapy, Rf-mediated activation of the cisplatin and carboplatin prodrugs cis,cis,trans-[Pt(NH ) (Cl) (O CCH CH CO H) ] (1) and cis,cis,trans-[Pt(NH ) (CBDCA)(O CCH CH CO H) ] (2, where CBDCA = cyclobutane dicarboxylate) resulted in pronounced reduction of the cell viability, including under hypoxia conditions. Such photoactivation mode occurs to a considerable extent intracellularly, as demonstrated for 1 by uptake and cell viability experiments. Pt NMR, DNA binding studies using circular dichroism, mass spectrometry and immunofluorescence microscopy were performed using the Rf-1 catalyst-substrate pair and indicated that cell death is associated with the efficient light-induced formation of cisplatin. Accordingly, Western blot analysis revealed signs of DNA damage and activation of cell death pathways through Rf-mediated photochemical activation. Phosphorylation of H AX as indicator for DNA damage, was detected for Rf-1 in a strictly light-dependent fashion while in case of free cisplatin also in the dark. Photochemical induction of nuclear pH AX foci by Rf-1 was confirmed in fluorescence microscopy again proving efficient light-induced cisplatin release from the prodrug system.