Serum‐Based Detection of Liver Pathology Using a Fluorogenic Alkaline Phosphatase Probe

Abstract

Development of “ultrahigh contrast” fluorogenic probes for trapping alkaline phosphatase (ALP) activities in human serum is highly desirable for clinical auxiliary diagnosis for hepatobiliary diseases. However, the intrinsic dilemma of incomplete ionization of intramolecular charge transfer (ICT)‐based ALP fluorophores and autofluorescence interference of serum result in low sensitivity and accuracy. Given that unique halogen effects could lead to a drastic decrease in the pKa value and a significant enhancement in the fluorescence quantum yield, herein we report an enzyme‐activatable near‐infrared probe based on a difluoro‐substituted dicyanomethylene‐4H‐chromenep for achieving fluorescent quantification of human serum ALP. Rational design strategy is demonstrated by altering the substituted halogen groups to well regulate the pKa for meeting the physiological precondition. Owing to the complete ionization at pH 7.4 with tremendous fluorescence enhancement, the difluoro‐substituted DCM‐2F‐HP manifests a linear relationship between the emission intensity and ALP concentration in both solution and serum samples. Along with measuring 77 human serum samples, the DCM‐2F‐HP based fluorescence method not only exhibits significant correlations with clinical colorimetry, but also distinguishes ALP patients from healthy volunteers, as well as assessing the progress of liver disease, thus providing a potential toolbox for quantitatively detecting ALP and warning the stage of hepatopathy.

Activatable Near‐Infrared Versatile Fluorescent and Chemiluminescent Dyes Based on the Dicyanomethylene‐4H‐pyran Scaffold: From Design to Imaging and Theranostics

“…Recently, Zhu's group further reduced the pK a value of the DCM fluorophore to 6.0 through a difluoro-substituted approach, resulting in enhancing its fluorescence efficiency in the biological pH range. [83] Then, the difluoro-substituted dye was rationally developed as an ALP-responsive NIR fluorescent dye, DCM-2F-HP, for diagnosing earlystage liver disease by measuring ALP levels.…”

“…Subsequently, a NIR light‐responsive nanocontainer loaded with Cl 2 ‐BDCM‐ALP has been readily constructed and employed for time‐resolved imaging of ALP activity in vivo for the first time. Recently, Zhu's group further reduced the p K a value of the DCM fluorophore to 6.0 through a difluoro‐substituted approach, resulting in enhancing its fluorescence efficiency in the biological pH range [83] . Then, the difluoro‐substituted dye was rationally developed as an ALP‐responsive NIR fluorescent dye, DCM‐2F‐HP , for diagnosing early‐stage liver disease by measuring ALP levels.…”

A Near-Infrared Fluorescent Dye with Tunable Emission Wavelength and Stokes Shift as a High-Sensitivity Cysteine Nanoprobe for Monitoring Ischemic Stroke

“…Dicyanomethylene-4 H -chromene (DCM) derivatives have been a hotspot of research in recent years, especially in the fields of fluorescent probes and nanodrug delivery. − DCM derivatives are a class of dyes designed based on the intramolecular charge transfer (ICT) mechanism. , In recent years, the electron donor and electron acceptor in DCM’s structures have been designed to achieve major breakthroughs in emission wavelength, Stokes shift, and fluorescence quantum yield. − There are four kinds of electron acceptor skeletons, among which oxygen-substituted and nitrogen-substituted skeletons are widely investigated. , The sulfur-substituted and selenium-substituted skeletons have been rarely reported. However, when the electron donor is consistent, it has been reported that sulfur-substituted and selenium-substituted skeletons have advantages over oxygen-substituted and nitrogen-substituted skeletons in terms of both emission wavelength and Stokes shift .…”