Copper(II) complexes of N3O tripodal ligands appended with pyrene and polyamine groups: Anti-proliferative and nuclease activities

Abstract

A series of tripodal ligands based on the 2-tert-butyl-4-R-6-phenol was synthesized, where R=aldehyde (HL¹), R=putrescine-pyrene (HL²) and R=putrescine (HL³). A dinucleating ligand wherein a putrescine group connects two tripodal moieties was also prepared (H₂L⁴). The corresponding copper complexes (1, 2, 3, and 4, respectively) were prepared and characterized. We determined the phenol's pKas in the range 2.47-3.93. The DNA binding constants were determined at 6×10⁶, 5.5×10⁵ and 2.7×10⁶ for 2, 3 and 4, respectively. The complexes display a metal-centered reduction wave at E_p^c,red=-0.45 to -0.5V vs. saturated calomel electrode, as well as a ligand-centered oxidation wave above 0.57V at pH7. In the presence of ascorbate they promote an efficient cleavage of DNA, with for example a concentration required to cleave 50% of supercoiled DNA of 1.7μM for 2. The nuclease activity is affected by the nature of the R group: putrescine-pyrene≈bis-ligating>putrescine>aldehyde. The species responsible for strand scission is the hydroxyl radical. The cytotoxicity of the complexes was evaluated on bladder cancer cell lines sensitive or resistant to cis-platin. The IC₅₀ of complexes 2 and 4 span over a short range (1.3-2μM) for the two cell lines. They are lower than those of the other complexes (3.1-9.7μM) and cis-platin. The most active compounds block the cell cycle at the G0/1 phase and promote apoptosis.

Keywords: Anti-tumor; Copper; DNA; Nuclease; Phenol; Tripodal ligand.