Molecular Design Strategy to Construct the Near-Infrared Fluorescent Probe for Selectively Sensing Human Cytochrome P450 2J2

Abstract

Cytochrome P450 2J2 (CYP2J2), a key enzyme responsible for oxidative metabolism of various xenobiotics and endogenous compounds, participates in a diverse array of physiological and pathological processes in humans. Its biological role in tumorigenesis and cancer diagnosis remains poorly understood, owing to the lack of molecular tools suitable for real-time monitoring CYP2J2 in complex biological systems. Using molecular design principles, we were able to modify the distance between the catalytic unit and metabolic recognition moiety, allowing us to develop a CYP2J2 selective fluorescent probe using a near-infrared fluorophore (E)-2-(2-(6-hydroxy-2, 3dihydro-1H-xanthen-4-yl)vinyl)-3,3-dimethyl-1-propyl-3H-indol-1-ium iodide (HXPI). To improve the reactivity and isoform specificity, a self-immolative linker was introduced to the HXPI derivatives in order to better fit the narrow substrate channel of CYP2J2, the modification effectively shortened the spatial distance between the metabolic moiety (O-alkyl group) and catalytic center of CYP2J2. After screening a panel of O-alkylated HXPI derivatives, BnXPI displayed the best combination of specificity, sensitivity and applicability for detecting CYP2J2 in vitro and in vivo. Upon O-demethylation by CYP2J2, a self-immolative reaction occurred spontaneously via 1,6-elimination of phydroxybenzyl resulting in the release of HXPI. Allowing BnXPI to be successfully used to monitor CYP2J2 activity in real-time for various living systems including cells, tumor tissues, and tumor-bearing animals. In summary, our practical strategy could help the development of a highly specific and broadly applicable tool for monitoring CYP2J2, which offers great promise for exploring the biological functions of CYP2J2 in tumorigenesis.

A Simple Near‐Infrared Fluorescent Probe for the Detection of Peroxynitrite

“…Not only does NIR fluorescence avoid interference with the auto‐fluorescence of indigenous molecules, near‐infrared light results in less scattering and deeper tissue penetration, limiting the damage to living cells . As such, the development of NIR fluorescent sensors is of growing interest to the sensing community …”

Semi-Quantitatively Designing Two-Photon High-Performance Fluorescent Probes for Glutathione S-Transferases

“…Recently, small-molecule fluorescent probes have been rapidly emerging as a powerful tool for enzyme detection in biological samples by virtue of their fast analysis, higher sensitivity, minimal perturbation to living systems, and real-time detection capabilities [9][10][11][12][13]. Indeed, several such probes have been developed for sensitive detection of GST activities with representatives being DNAT-Me [14], DNs-CV [15], and 3,4-DNADCF [16].…”

The cross-talk modulation of excited state electron transfer to reduce the false negative background for high fidelity imaging in vivo

“…Owing to their reactive specicity, timely reporting and biological affinity, molecular-probe-based uorescence imaging techniques have been rapidly developed for biological monitoring, molecular medicine and even surgical navigation. [1][2][3][4][5][6][7][8][9][10] However, in the actual imaging, there are several problems that are usually neglected: (1) inherent false negative background (FNB): although the background seems to be dark, the real background uorescence will be exposed when the intensity scale of microscopy is compressed (Scheme 1A). Obviously, the real background is covered by the wide intensity scale of microscopy.…”

“…Accordingly, we constructed a system of a uorophore (F)electron donor (D)-electron regulator (R), and established a dual modulation strategy for the de novo design of high-delity imaging probes based on the cross-talk effect of ESET and photo induced electron transfer (PET). F-D-R worked as follows: (1) ensuring PET for uorescence quenching and adjusting R for enhanced ESET from F to D, to minimize the FNB; and 2inhibiting PET for uorescence restoration and decreasing the ESET from F to D, to enhance the uorescence, thereby further minimizing the FNB. For the implementation, we synthesized 7hydroxy-2-oxo-2H-chromene-3-carbaldehyde (HPC)-n series probes, consisting of the uorophore 7-hydroxy-2-oxo-2Htryptone-3-acetaldehyde (F), an adjustable electron-rich donor 2-hydrazinopyridine (D), as well as the electronic regulators (R).…”