This study reports the solvent-free mechanochemical synthesis of a novel series of 2-hydrazone-bridged benzothiazole derivatives
19–
52 via the reaction of 2-hydrazinylbenzothiazole derivatives
4–
6 with
O-alkylated benzaldehydes
7–
18. The stereostructure of the
E-isomers was confirmed by 2D NOESY spectroscopy. The antiproliferative potential of these newly prepared 2-hydrazone derivatives of benzothiazole
19–
52 was evaluated in vitro against eight human cancer cell lines. Several compounds demonstrated low micromolar IC
50 values, with some outperforming the reference drug etoposide. Among the most potent compounds, the 6-chloro-2-hydrazone(3-fluorophenyl)benzothiazole derivative
38 exhibited remarkable activity against pancreatic adenocarcinoma (Capan-1, IC
50 = 0.6 µM) and non-small cell lung cancer (NCI-H460, IC
50 = 0.9 µM). Structure–activity relationship analysis revealed that derivatives
45–
52, featuring a methoxy group at position 6 of the benzothiazole ring and either a methoxy or fluorine substituent at position 3 of the phenyl ring, showed consistently strong antiproliferative effects across all tested cell lines (IC
50 = 1.3–12.8 µM). Furthermore, compounds bearing
N,
N-diethylamino or
N,
N-dimethylamino groups at position 4 of the phenyl ring generally exhibited superior activity compared to those with morpholine or piperidine moieties. However, as this study represents an initial screening, further mechanistic investigations are required to confirm specific anticancer pathways and therapeutic relevance. In addition to their in vitro anticancer properties, the antibacterial activity of the compounds was assessed against both Gram-positive and Gram-negative bacteria. Notably, compound
37 demonstrated selective antibacterial activity against
Pseudomonas aeruginosa (MIC = 4 µg/mL). Overall, this work highlights the efficiency of a green, mechanochemical approach for synthesizing
E-isomer hydrazone-bridged benzothiazoles and underscores their potential as promising scaffolds for the development of potent antiproliferative agents.
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