Coordination geometry-induced optical imaging of L-cysteine in cancer cells using imidazopyridine-based copper(II) complexes

Overexpression of cysteine cathepsins proteases has been documented in a wide variety of cancers, and enhances the L-cysteine concentration in tumor cells. We report the synthesis and characterization of copper(II) complexes [Cu(L1)2(H2O)](SO3CF3)2, 1, L1 = 3-phenyl-1-(pyridin-2-yl)imidazo[1,5-a]pyridine, [Cu(L2)2(SO3CF3)]SO3CF3, 2, L2 = 3-(4-methoxyphenyl)-1-pyridin-2-yl-imidazo[1,5-a]pyridine, [Cu(L3)2(H2O)](SO3CF3)2, 3, L3 = 3-(3,4-dimethoxy-phenyl)-1-pyridin-2-yl-imidazo[1,5-a]pyridine and [Cu(L4)2(H2O)](SO3CF3)2, 4, L4 = dimethyl-[4-(1-pyridin-2-yl-imidazo[1,5-a]pyridin-3-yl)phenyl]amine as ‘turn-on’ optical imaging probes for L-cysteine in cancer cells. The molecular structure of complexes adopted distorted trigonal pyramidal geometry (τ, 0.68–0.87). Cu–Npy bonds (1.964–1.989 A) were shorter than Cu–Nimi bonds (2.024–2.074 A) for all complexes. Geometrical distortion was strongly revealed in EPR spectra, showing g‖ (2.26–2.28) and A‖ values (139–163 × 10−4 cm−1) at 70 K. The d–d transitions appeared around 680–741 and 882–932 nm in HEPES, which supported the existence of five-coordinate geometry in solution. The Cu(II)/Cu(I) redox potential of 1 (0.221 V vs. NHE) was almost identical to that of 2 and 3 but lower than that of 4 (0.525 V vs. NHE) in HEPES buffer. The complexes were almost non-emissive in nature, but became emissive by the interaction of L-cysteine in 100% HEPES at pH 7.34 via reduction of Cu(II) to Cu(I). Among the probes, probe 2 showed selective and efficient turn-on fluorescence behavior towards L-cysteine over natural amino acids with a limit of detection of 9.9 × 10−8 M and binding constant of 2.3 × 105 M−1. The selectivity of 2 may have originated from a nearly perfect trigonal plane adopted around a copper(II) center (∼120.70°), which required minimum structural change during the reduction of Cu(II) to Cu(I) while imaging Cys. The other complexes, with their distorted trigonal planes, required more reorganizational energy, which resulted in poor selectivity. Probe 2 was employed for optical imaging of L-cysteine in HeLa cells and macrophages. It exhibited brighter fluorescent images by visualizing Cys at pH 7.34 and 37 °C. It showed relatively less toxicity for these cell lines as ascertained by the MTT assay.