Design and in vitro anticancer assessment of a click chemistry-derived dinuclear copper artificial metallo-nuclease
Journal article, 2025
Copper compounds with artificial metallo-nuclease (AMN) activity are mechanistically unique compared to established metallodrugs. Here, we describe the development of a new dinuclear copper AMN, Cu2-BPL-C6 (BPL-C6 = bis-1,10-phenanthroline-carbon-6), prepared using click chemistry that demonstrates site-specific DNA recognition with low micromolar cleavage activity. The BPL-C6 ligand was designed to force two redox-active copper centres - central for enhancing AMN activity - to bind DNA, via two phenanthroline ligands separated by an aliphatic linker. DNA-binding experiments, involving circular dichroism spectroscopy, agarose gel electrophoresis and fluorescence quenching, revealed a preference for binding with adenine-thymine-rich DNA. The oxidative cleavage mechanism of Cu2-BPL-C6 was then elucidated using in vitro molecular and biophysical assays, including in-liquid atomic force microscopy analysis, revealing potent DNA cleavage mediated via superoxide and hydrogen peroxide oxidative pathways. Single-molecule analysis with peripheral blood mononuclear cells identified upregulated single-strand DNA lesions in Cu2-BPL-C6-treated cells. Using specific base excision repair (BER) enzymes, we showed that Endo IV selectively repairs these lesions indicating that the complex generates apurinic and apyrimidinic adducts. Broad spectrum anticancer evaluation of BPL-C6 was performed by the National Cancer Institute's 60 human cell line screen (NCI-60) and revealed selectivity for certain melanoma, breast, colon and non-small cell lung cancer cell lines.
Author
Simon Poole
Dublin City University
Obed Akwasi Aning
Chalmers, Life Sciences, Chemical Biology
Vickie McKee
University of Southern Denmark
Dublin City University
Thomas Catley
University of Sheffield
Aaraby Yoheswaran Nielsen
Odense Universitetshospital
Helge Thisgaard
Odense Universitetshospital
University of Southern Denmark
Pegah Johansson
Sahlgrenska University Hospital
University of Gothenburg
Georgia Menounou
Dublin City University
Joseph Hennessy
Dublin City University
Creina Slator
Dublin City University
Alex Gibney
Dublin City University
Alice Pyne
University of Sheffield
Bríonna McGorman
Dublin City University
Fredrik Westerlund
Chalmers, Life Sciences, Chemical Biology
Andrew Kellett
Dublin City University
Nucleic Acids Research
0305-1048 (ISSN) 1362-4962 (eISSN)
Vol. 53 1 gkae1250Subject Categories (SSIF 2025)
Molecular Biology
Medical Biotechnology
DOI
10.1093/nar/gkae1250
Related datasets
AFM data for: Development and In Vitro Anticancer Evaluation of a Click Chemistry Derived Dinuclear Copper Artificial Metallo-nuclease. [dataset]
DOI: 10.15131/shef.data.27613857