Dual inhibition of AChE and BuChE by coumarin–benzothiazole hybrids: potent candidates unveiled through innovative screening and docking studies

= 0.9983-0.9992), precision (CV ≤ 7.8%), and accuracy within accepted validation limits. Biological evaluation revealed highly potent inhibitors, particularly 13a and 13m, which exhibited IC₅₀ values of 1.0 ± 0.1 nM and 0.6 ± 0.1 nM against AChE, respectively, compared with 64.9 ± 16.0 nM for donepezil. For BuChE, 13m showed an IC₅₀ of 109.2 ± 14.7 nM, while 13a displayed 1212.5 ± 144.0 nM, both lower than donepezil (2202 ± 326 nM). Kinetic studies confirmed a competitive mechanism of inhibition, with Kᵢ values as low as 0.12 ± 0.02 nM for AChE and 47.76 ± 15.14 nM for BuChE (13m). Molecular docking studies supported these findings, demonstrating key interactions within the catalytic sites of both enzymes. Additionally, in silico evaluations indicated that these derivatives have favorable druglikeness. Together, the results highlight coumarin-benzothiazole derivatives as promising dual-target ChEIs and establish the developed analytical method as a versatile tool for the discovery and characterization of new anti-AD candidates.