Tang_2025_Spectrochim.Acta.A.Mol.Biomol.Spectrosc_349_127342

In this work, a series of fluorescent probes were designed and synthesized based on the consideration of structural differences between acetylcholinesterase and butyrylcholinesterase(BChE) in the activity pockets, using hemicarbocyanine fluorophore as the parent skeleton, with p-aminobenzenesulfonamide incorporated as the Golgi localization group. Among them, the probe CYB-B4 containing acylcyclohexanes as the recognizing group, exhibiting absorption/emission wavelengths of 675 nm/710 nm respectively, demonstrated high sensitivity toward BChE, with a detection limit as low as 2.2 mU/mL. The probe showed rapid response to BChE, reaching complete response within 36 min; moreover, CYB-B4 could effectively minimize the interference from acetylcholinesterase(AChE), and displayed relatively low biotoxicity. The cell imaging results demonstrated that the probe CYB-B4 possessed Golgi co-localization ability with a co-localization coefficient of 0.75, and could be successfully applied to the detection of endogenous butyrylcholinesterase in cells.