Design, Synthesis, and Biological Evaluation of Coumarin Derivatives Tethered to an Edrophonium‐like Fragment as Highly Potent and Selective Dual Binding Site Acetylcholinesterase Inhibitors

Abstract A large series of substituted coumarins linked through an appropriate spacer to 3‐hydroxy‐ N , N ‐dimethylanilino or 3‐hydroxy‐ N , N , N ‐trialkylbenzaminium moieties were synthesized and evaluated as acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) inhibitors. The highest AChE inhibitory potency in the 3‐hydroxy‐ N , N ‐dimethylanilino series was observed with a 6,7‐dimethoxy‐3‐substituted coumarin derivative, which, along with an outstanding affinity (IC 50 =0.236 n M ) exhibits excellent AChE/BChE selectivity (SI>300 000). Most of the synthesized 3‐hydroxy‐ N , N , N ‐trialkylbenzaminium salts display an AChE affinity in the sub‐nanomolar to picomolar range along with excellent AChE/BChE selectivities (SI values up to 138 333). The combined use of docking and molecular dynamics simulations permitted us to shed light on the observed structure–affinity and structure–selectivity relationships, to detect two possible alternative binding modes, and to assess the critical role of π–π stacking interactions in the AChE peripheral binding site.