Discovery of new thiazolyl hydrazone derivatives as anti-breast cancer agents: Synthesis, biological evaluation and in silico studies.

A series of new compounds was designed and synthesized based on the pronounced anticancer activities of the thiazole ring and hydrazone groups against breast cancer cells. IR, 1H NMR, 13C NMR (decoupled and APT), 19F NMR, 2D NMR (HSQC), HRMS and elemental analysis data were used to confirm the structures of the compounds. Cytotoxicity was assessed by the MTT assay, while immunoperoxidase staining was used to examine the distribution of caspase-3, BAX, Bcl2, cytochrome c, Ki-67, RIPK1, RIPK3, and MLKL in breast cancer cell lines (M4A4 and MCF-7). Compounds 4b, 4c, 4d, 4g, 4h, and 4l were the most effective derivatives in the series against MCF-7 and M4A4 cancer cells, with IC50 values of 51-106 μM and 12.87-100 μM, respectively. Molecular docking and molecular dynamics (MD) simulations were performed to elucidate the interactions between compound 4l, which has the best BAX: Bcl2 ratio, and the BAX and Bcl2 proteins. In addition, molecular docking and MD simulations showed that compound 4l stably binds within the Bcl2 BH3-binding groove, maintaining key interactions with ASP108, TYR105, and ARG143 during the 300 ns simulation. The RMSD remained below 2.5 Å after equilibration, supporting the structural stability of the complex.