数字农科院2.0

An acetylcholinesterase-insensitive near-infrared fluorescent probe for specific and real-time detection of butyrylcholinesterase in vitro and in vivo

文献类型： 外文期刊

作者： Qi Sun; Getong Zhang; Ting Zhang; Yuan Qiu; Jingfang Yang; Xiaogang Luo; Genyan Liu

关键词： Butyrylcholinesterase; Near-infrared; Fluorescent probe; Inhibitor screening

期刊名称： MICROCHEMICAL JOURNAL

ISSN： 0026-265X

年卷期： 2025 年

页码：

收录情况： SCIE（2025版）

摘要： Butylcholinesterase (BChE) is a crucial enzyme intricately linked to liver and neurological diseases. Accurate monitoring of BChE activity in complex biological systems is essential for understanding its physiological and diagnostic roles. However, this is often hampered by cross-reactivity with the structurally similar acetylcholin-esterase (AChE). To address this, we developed Cy-BChE, a near-infrared ffuorescent probe, that integrates a Cy-NIR-OH ffuorophore with a cyclopropyl methyl ester recognition moiety for the speciffc and real-time detection of BChE both in vitro and in vivo. Cy-BChE features favorable optical properties, including long excitation/ emission wavelengths (λex = 670 nm/λem = 720 nm) for deep tissue penetration and minimal background, a rapid ffuorescence response (25 min), and an excellent detection limit (2.08 × 10− 7 U/mL). Kinetic analysis conffrmed its strong enzyme-probe afffnity (Km = 0.16 ± 0.02 μM), supporting its use for quantitative analysis. Moreover, Cy-BChE’s turn-on response enables sensitive evaluation of BChE inhibitors, quantifying IC50 value of rivastigmine and tacrine as 10.86 ± 1.72 μM and 53.86 ± 1.02 nM, respectively. Most importantly, Cy-BChE shows minimal AChE interference, ensuring highly BChE selectivity. We also demonstrated its good biocom-patibility, low cytotoxicity and ability for real-time imaging of endogenous BChE in live cells and zebraffsh. Overall, Cy-BChE provides a powerful tool for speciffc BChE detection, inhibitor screening and real-time tracking in complex biological environments, with excellent AChE discrimination.

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