Euparin and 2‑hydroxy acetophenone hydrazone derivatives (2–7) were synthesized, and their structure was characterized by Fourier Transform Infrared (FT-IR), Nuclear Magnetic Resonance (NMR), and Mass (MS) spectral data. In vitro, using a modified Ellman's spectrophotometric approach, the inhibitory potential of these substances on the AchE (Acetylcholinesterase) enzyme was determined. Compounds 3, 5, and 7 exhibited strong and specific inhibitors of AchE. Ortho hydroxy acetophenone hydrazones were less effective than the euparin-based compounds 5 and 7, with an IC50 of 22 nM and 14 nM, respectively. The highest AchE inhibitory activity was seen in compound 7, which was close to what was observed for the reference drug, Donepezil. To further investigate the potential interactions between these chemicals and AchE, docking tests were conducted. A link between the biological, electronic, and physicochemical credentials of the compounds in the title was established through structure-activity relationships and in silico ADME studies. Molecular docking simulation disclosed that the highest docking scores were -9.96 and -10.77 kcal/mol for compounds 5 and 7, respectively. Further, the antibacterial and antifungal activity of samples 1–7 were evaluated against Candida albicans, Gram-positive and Gram-negative bacteria.