Synthesis, computational analysis, and anti-proliferative evaluation of novel bithienylbenzamidine derivatives: Towards cytostatic cancer therapeutics

• Synthesis of four novel monocationic 2,5-diarylbithiophene derivatives via innovative Suzuki coupling reactions. • Advanced DFT calculations revealing crucial electronic properties and structure-activity relationships. • Potent anti-proliferative effects across diverse cancer types demonstrated through rigorous NCI-60 screening. • Exceptional growth inhibition by compound 4c with GI50 of 0.36 µM against specific ovarian cancer lines. • Potential mechanism targeting human carbonic anhydrase IX in cancer cells suggested by molecular docking. Our research unveils the synthesis and characterization of four novel monocationic 2,5-diarylbithiophene derivatives 4a-d , prepared through Suzuki cross-coupling reactions and subsequent amidine formation. Density Functional Theory (DFT) calculations provide unprecedented insights into the electronic properties and structure-activity relationships of these compounds. In vitro screening against the NCI-60 cancer cell panel demonstrates remarkable anti-proliferative activities. Compound 4c emerges as a standout candidate, exhibiting potent growth inhibition across multiple cancer types, with GI 50 values as low as 0.36 µM against certain ovarian cancer lines. Intriguingly, our research suggests a potential shift from purely cytotoxic towards more cytostatic behavior, potentially offering improved therapeutic windows and reduced side effects. Molecular docking studies provide evidence for the compounds' interaction with human carbonic anhydrase IX, hinting at a possible mechanism of action. This comprehensive study not only introduces a promising new class of anti-cancer compounds but also provides a rich foundation of structural, electronic, and biological data to fuel future research. The bithienylbenzamidine derivatives, particularly compound 4c , represent exciting candidates for further development in the ongoing battle against cancer.