Design, Discovery, Synthesis, Molecular Docking, MD Simulation, and In Vitro Biological Evaluation of Thiazolidinedione-Clubbed Benzimidazole Hybrids as Potential Anticancer Agents.

Abstract

Introduction/objective: Breast cancer is a major global health issue. A new series of thiazolidinedione-benzimidazole hybrid derivatives (TBZ1-15) was developed and tested for anticancer properties against the MCF-7 breast cancer cell line using the MTT assay. Among these compounds, TBZ-13 showed the greatest cytotoxicity, with an IC₂⁽ of 35 μM.

Method: To further elucidate its mechanism of action, TBZ-13 was subjected to multiple in vitro assays, including colony formation, Hoechst staining, reactive oxygen species (ROS) generation, cell migration, and propidium iodide (PI) staining. The results demonstrated that TBZ- 13 significantly inhibits clonogenic survival and cell migration, while inducing pronounced nuclear condensation and fragmentation, confirming apoptosis. Moreover, TBZ-13 caused a marked increase in intracellular ROS levels, indicating that oxidative stress-mediated cytotoxicity is a leading mechanism of cytotoxicity.

Results and discussion: Notably, TBZ-13 outperformed the reference drug troglitazone in suppressing metastatic potential and promoting cancer cell death. Molecular docking studies were conducted to investigate interactions with peroxisome proliferator-activated receptor gamma (PPAR-γ), a key target implicated in tumor progression, angiogenesis, and metastasis. The crystal structure of human PPAR-γ (PDB ID: 1FM9; resolution 2.10 Å) was used for docking analysis. TBZ-13 displayed favorable binding conformations with critical hydrogen bonding interactions involving Tyr473, His323, and Ser289. Overall, these findings suggest that TBZ-13 is a promising anticancer lead candidate for further development.

Conclusion: The study focuses on breast cancer, a leading cause of death globally, introducing a series of thiazolidinedione-benzimidazole hybrid derivatives (TBZ1-15) evaluated for anticancer activity against the MCF-7 cell line. TBZ-13 showcased the highest cytotoxicity with an IC₂⁽ of 35 μM. Subsequent in vitro assays revealed that TBZ-13 inhibits cell migration and clonogenic survival, induces apoptosis through nuclear condensation, and increases reactive oxygen species (ROS), indicating oxidative stress as its cytotoxic mechanism.