Alkaline phosphatase activatable near-infrared fluorescent probe for in-situ diagnosis of cholestatic liver injury

Abstract

Cholestatic liver injury (CLI) is a clinical syndrome caused by impaired bile excretion and is almost asymptomatic in early-stage. If CLI is not recognized timely, it will elicit more severe liver diseases. Therefore, earlier diagnosis of CLI is urgently needed but remains challenging. On the basis of the consensus that elevation of serum alkaline phosphatase (ALP) is the most susceptible indicator of earlier cholestatic diseases. Herein, we conjugated dicyanoisophorone with hydroxyl-substituted coumarin derivative to afford an intramolecular charge transfer (ICT)-based fluorophore TX-OH, and successfully constructed an enzyme activatable fluorescent probe TX-PS, which enables specifically detecting ALP with near-infrared emission (720nm), large stokes shift (198nm) and excellent linear relationship (R2 = 0.9969) between fluorescence intensity and the concentrations of ALP (0-100U/L). Moreover, TX-PS exhibits applicable for imaging of ALP activities in living cells and zebrafish with ignorable toxicity and shows great potential to visually screen ALP inhibitors. Importantly, the liver tissues from CLI mice displayed remarkably higher fluorescence intensity than that from normal mice, suggesting the valuable capability of TX-PS for evaluating CLI degree. Overall, this work affords a powerful tool for earlier diagnosis of CLI and high-throughput screening of ALP inhibitors or new drugs for CLI therapeutics.