Synthesis of novel thiophene derivatives: Evaluation of antioxidant, anticancer, and AChE inhibitory activities

Background: Heterocyclic compounds hold a significant position in pharmaceutical chemistry owing to their extensive range of biological activities. Objective: It was aimed to investigate the antioxidant, anticancer and antimicrobial activities of two newly synthesized thiophene derivatives (3 and 5), their basic ADME/T properties in silico and their effects on the acetylcholinesterase enzyme in vitro and in silico. Methods: Thiophene derivatives were synthesized via Sonogashira Coupling reaction. These derivatives were characterized using HRMS, 1H NMR and 13C NMR. The biological evaluation encompassed antioxidant, antimicrobial, and apoptotic assays, as well as enzyme inhibition studies. Computational analysis included molecular docking and ADME/T profiling. Results: Compound 3 exhibited significantly high cytotoxic activity than 5 on A549, Caco-2 and HepG2 cancer cell lines, except LNCaP. Similarly, Compound 3 exhibited better apoptotic activity than 5 in all cancer cell lines when compared by the Annexin V-FITC/PI method. Furthermore, 3 showed better apoptotic activity than paclitaxel in the LNCaP cell line. qPCR analysis showed that 3 and 5 upregulated proapoptotic genes (BAX, CASP3, CASP8 and CASP9) and downregulated antiapoptotic gene BCL2. ADME analysis showed that both thiophene derivatives passed Lipinski's rule of five and could pass almost freely through the gastrointestinal and especially the Blood-Brain Barrier. In addition, 3 was more embedded in the narrow gorge formed by the catalytic triad in the active site (SER203-GLU334-HIS447) than 5 by molecular docking analysis. Conclusion: Consequently, 3, which was synthesized for the first time and whose biological properties were investigated within the scope of this study, seems to have potential in both cancer and Alzheimer's disease treatment.