Diketopyrrolopyrrole-based fluorescence probes for the imaging of lysosomal Zn²⁺ and identification of prostate cancer in human tissue

Abstract

A series of diketopyrrolopyrrole-based fluorescent probes (<b>DPP-C2</b>, <b>LysoDPP-C2</b>, <b>LysoDPP-C3</b>, and <b>LysoDPP-C4</b>) have been developed for the detection of low pH and Zn²⁺ in an AND logic fashion. The chelation of Zn²⁺ or the protonation of a morpholine moiety within these probes results in a partial increase in the fluorescence intensity, an effect ascribed to suppression of one possible photo-induced electron transfer (PET) pathway. In contrast, a large increase in the observed fluorescence intensity is observed at low pH and in the presence of Zn²⁺; this is rationalized in terms of both possible PET pathways within the probes being blocked. Job plots, fluorescence titration curves, and isothermal titration calorimetry proved consistent with a 1 : 1 Zn²⁺ complexation stoichiometry. Each probe demonstrated an excellent selectivity towards Zn²⁺ and the resulting Zn²⁺ complexes demonstrated pH sensitivity over the 3.5-9 pH range. Fluorescence imaging experiments confirmed that <b>LysoDPP-C4</b> was capable of imaging lysosomal Zn²⁺ in live cells. Little evidence of cytotoxicity was seen. <b>LysoDPP-C4</b> was successfully applied to the bioimaging of nude mice, wherein it was shown capable of imaging the prostate. Histological studies using a human sample revealed that <b>LysoDPP-C4</b> can discriminate cancerous prostate tissue from healthy prostate tissue.