Pyrene-appended bipyridine hydrazone ligand as a turn-on sensor for Cu2+ and its bioimaging application

Abstract

A pyrene-appended bipyridine hydrazone-based ligand, HL, was synthesized and characterized by spectroscopic methods. Upon complexation with Cu(II), HL formed a hexanuclear paddlewheel metal–organic macrocycle (MOM) via self-assembly with a high association constant with the molecular formula of [Cu₆L₆(NO₃)₆]. Intermolecular and intramolecular π–π interactions were demonstrated in this hexanuclear Cu(II) complex. Further, it was observed that HL had the potential to detect a trace level of Cu(II) ion selectively among a wide range of biologically relevant metal ions in aqueous medium at physiological pH. Using HL, it was feasible to sense copper(II) ions in living cells due to its good cell permeability and high solubility under physiological conditions along with its high IC₅₀ value. The low detection limit, high sensitivity and good reproducibility make this Cu–sensor very promising. The complex (MOM) formed between the ligand and Cu(II) was found to be 1 : 1 on the basis of fluorescence titrations and was confirmed by ESI-MS. Moreover, single-crystal study of the hexanuclear self-assembled fluorescent species provided better insight into its chemistry, e.g. coordination environment and binding mode, unlike most of the metal sensors due to the lack of a single-crystal structure of the metal sensor complex. Cytotoxicity assay and bioimaging were performed in living cells (Vero cells), giving green fluorescent images. Fluorescence lifetime measurements and theoretical calculations were carried out. The morphology and topographic details on the surface of the metal–organic macrocycle (MOM) were studied by field-emission scanning electron microscopy (FESEM).