Synthesis, Biological Evaluation, and In Silico Studies of New Hydrazone-Sulfonate Hybrids as Dual Inhibitors of Carbonic Anhydrase and Cholinesterase Enzymes

In this study, we synthesized two novel hydrazones (2a–f and 3a–f) and investigated their inhibitory properties against human carbonic anhydrase isoenzymes I and II (hCA I and hCA II), and acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) by in vitro assay and in silico approaches. All tested compounds indicated nanomolar inhibition with IC₅₀ values in the range of 47.3–317.8 nM against hCA I, 71.9–376.2 nM against hCA II, 81.01–832.9 nM against AChE, and 134.5–2264 nM against BChE. Among the tested compounds, compounds 2a and 2b inhibited hCA I better compared to acetazolamide (AZA). All the molecules inhibited AChE better than rivastigmine. Compound 2f was the best inhibitor candidate for AChE. Compared to rivastigmine, some of the tested compounds (2b, 2c, 2f, 3a, and 3b) showed higher nanomolar inhibition against BChE. Compound 2b was the best multi-target inhibitor candidate against hCA I, hCA II, and BChE. Furthermore, B3LYP-D3BJ/6-311++G(d,p) method was used for geometric optimization and investigation of electronic properties of the compounds. Molecular docking and molecular dynamics simulations were applied to determine possible binding sites and analyze the interaction dynamics. Additionally, ADME, pharmacokinetic properties, and toxicological evaluation results of 2b and 2f were analyzed by SwissADME and ADMETlab 3.0.