Design, Synthesis, and In Vitro Biological Assessment of Novel Benzimidazole and Hydrazone Derivatives Against Pancreatic Cancer and Melanoma Cells

ABSTRACT

In this study, novel Benzimidazole‐Hydrazone derivatives ( 5A–5J ) were synthesized and evaluated for their in vitro anticancer activities against Panc‐1, SK‐MEL‐30, HeLa, and BEAS‐2B cell lines. WST‐1 cytotoxicity assays revealed that compound 5A displayed notable potency in SK‐MEL‐30 cells (IC ₅₀  = 23.25 ± 1.40 μM), whereas compound 5H exhibited significant cytotoxicity in Panc‐1 cells (IC ₅₀  = 27.6 ± 1.1 μM), with minimal effects on BEAS‐2B cells (> 200 μM), suggesting a favorable in vitro safety profile under the tested conditions. Flow cytometric analyses demonstrated that 5A and 5H induced apoptosis and enhanced the apoptotic response when combined with Cisplatin, increasing apoptotic rates up to 80% in SK‐MEL‐30 and 90% in Panc‐1 cells, respectively. Cell cycle analysis revealed that compound 5A induced G1 arrest in SK‐MEL‐30 cells, while compound 5H caused G2/M arrest in Panc‐1 cells. Wound healing assays further confirmed the antimigratory potential of both compounds, alone and in combination with Cisplatin. In silico ADMET profiling provided preliminary insights into the pharmacokinetic properties of compounds 5A and 5H , indicating generally acceptable drug‐likeness parameters within the limitations of computational prediction. Molecular docking analyses suggested potential interactions with the caspase‐3 active site; however, these findings should be interpreted as supportive computational observations and do not provide direct evidence of a specific molecular mechanism underlying the observed apoptotic effects. These findings indicated that compounds 5A and 5H exhibited notable in vitro anticancer activity and may serve as potential candidates for further investigation.