LAUSR

A simple “click” protocol was employed in the quest of synthesizing 1,2,3‐triazole‐linked benzimidazoles as promising anticancer agents on various human cancer cell lines such as A549, HCT116, SK‐Mel‐28, HT‐29, and MCF‐7. Compound 12j demonstrated significant cytotoxic potential towards SK‐Mel‐28 cancer cells (IC50: 4.17 ± 0.09 µM) and displayed no cytotoxicity (IC50: > 100 µM) against normal human BEAS‐2B cells inferring its safety towards normal healthy cells. Further to comprehend the underlying apoptosis mechanisms, AO/EB, dichlorodihydrofluorescein diacetate (DCFDA), and 4',6‐diamidino‐2‐phenylindole (DAPI) staining were performed, which revealed the nuclear and morphological alterations. Compound 12j displayed impairment in cellular migration and inhibited colony formation. The annexin V binding assay and JC‐1 were implemented to evaluate the scope of apoptosis and the loss of the mitochondrial transmembrane potential in SK‐Mel‐28 cells. Cell‐cycle analysis revealed that compound 12j arrested the cells at the G2/M phase in a dose‐dependent manner. Target‐based assays established the inhibition of tubulin polymerization by 12j at an IC50 value of 5.65  ± 0.05 μM and its effective binding with circulating tumor DNA as a DNA intercalator. The detailed binding interactions of 12j with tubulin and DNA were examined by docking studies on PDB ID: 3E22 and DNA hexamer (PDB ID: 1NAB), respectively.