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Beyond Thioflavin-T: Excited State Intramolecular Proton Transfer-Active 2-Phenylbenzimidazoles for Sensitive and
Saddam Miya1, Tamanna Mallick1, Abhijit Karmakar1
1Department of Chemistry, Visva-Bharati (A Central University), Santiniketan, West Bengal 731235, India.
Abstract:
We introduce a novel series of excited state intramolecular proton transfer (ESIPT)-active 2-phenylbenzimidazole fluorophores (1-3) as highly selective turn-on probes for insulin amyloid aggregates (IAAs). Synthesized via an eco-friendly, metal-free route, these probes show high specificity for IAAs over native insulin and Thioflavin-T (ThT). Density functional theory (DFT) calculations confirm the intrinsic ESIPT mechanism, while photophysical studies reveal that a specific binding mode restricts molecular motion, enhancing quantum yield and fluorescence lifetime. Crucially, the lead compound (3) binds to a unique site distinct from ThT, as proven by competitive binding and Förster Resonance Energy Transfer (FRET) assays. It also exhibits superior affinity, a low detection limit, the ability to modulate aggregation kinetics, and excellent biocompatibility. This work moves beyond ThT, positioning the ESIPT-active 2-phenylbenzimidazole scaffold as a promising theranostic tool for amyloid-related pathologies such as Type-2 diabetes.
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