Design and in vitro evaluation of novel tetrazole derivatives of dianisidine as anticancer agents targeting Bcl-2 apoptosis regulator

This study examines the synthesis and biological evaluation of novel tetrazole derivatives of 3,3'-dimethoxybenzidine as potential Anticancer agents, focusing on their cytotoxic, apoptotic, and anti-inflammatory properties. Ten derivatives were synthesized using thioureas as precursors, characterized through spectroscopic methods, and assessed for their in silico toxicological profiles using the ADMET-AI and ProTox 3.0 platforms. In vitro cytotoxic activity was evaluated against four human Cancer cell lines (HTB-140, A549, HeLa, SW620) and one normal cell line (HaCaT) using MTT and LDH assays. Mechanistic studies included Apoptosis assessment via flow cytometry and interleukin-6 (IL-6) analysis using ELISA. The synthesized tetrazole derivatives demonstrated significant Anticancer potential, exhibiting selective cytotoxicity against Cancer cell lines, robust induction of Apoptosis, and a notable reduction in IL-6 levels. Their favorable toxicity profiles, as observed in both in silico and in vitro evaluations, support their potential as promising candidates for further development. The tested compounds showed strong inhibitory activity against the Apoptosis regulator Bcl-2, with binding affinities comparable to those of native ligands. Western blot analysis revealed a dramatic loss of Bcl-2 protein expression in selected Cancer cells during exposure to compound 5. Additionally, this research highlights the innovative use of hazardous substrates in drug discovery, aligning with the principles of green chemistry. Future efforts should focus on optimizing the most active derivatives and conducting in vivo studies to confirm their therapeutic potential and safety.

Anticancer; Apoptosis; Cytotoxicity; Tetrazoles; Toxicology.