Synthesis, antimicrobial, anticancer activity, 3D QSAR, ADMET properties, and in silico target fishing of novel N,N-disubstituted chloroacetamides

Abstract

The recent FDA approval of afatanib, ibrutinib, and osimertinib, which covalently bind to specific cysteine residues in target kinases, has renewed interest in covalent drug discovery. Besides α,β-unsaturated carbonyls, chloroacetamides have emerged as popular warheads for designing targeted covalent inhibitors. In this study, we synthesized thirteen N,N-disubstituted chloroacetamides (1–13) by acylating secondary amines with chloroacetyl chloride, selecting substituents to provide a wide range of lipophilicity. We evaluated their anticancer and antimicrobial activity, finding five compounds with significant cytotoxicity against HeLa, K562, and A549 cell lines (IC<inf>50</inf> <10 μM). Notably, compound 10 activated caspases 3, 8, and 9, promoting both intrinsic and extrinsic apoptotic pathways, while compounds 9–12 were strong apoptosis inducers. A 3D QSAR model showed that aromatic substituents on nitrogen atoms reduced HeLa potency, whereas the overall molecular shape had a positive effect. ChemBL and pharmacophoric similarity searches suggested potential anticancer targets, including alcohol dehydrogenase (ADH), aldehyde dehydrogenase (ALDH), glycogen synthase kinase-3β, and calmodulin. Docking studies indicated that chloroacetamides bind to ADH and ALDH via hydrogen bonds and hydrophobic interactions. Pharmacokinetics predictions suggest that chloroacetamides are druglike molecules with promising ADMET properties.