Isatin‐Acenaphthoquinone Derived <i>tetra</i> ‐Schiff Base: Synthesis, Molecular Docking, Molecular Dynamics Simulation and Cytotoxic Potential in MCF‐7 Breast Cancer Cells

Herein, isatin was derivatized into tetra-imine derivative (8d) using active reactants and their anticancer potential against human breast cancer cell (MCF-7), are reported. However, the analytical techniques named FT-IR, ¹H and ¹ ³C NMR, were utilized to confirming of the synthesized compounds. In vitro investigation, MTT assay was demonstrated a dose-dependent decrease in MCF-7 cell viability, with an IC₅₀ of 56 µM and 30% viability at 100 µM concentration as well. Morphological studies indicated apoptosis as the primary mechanism, evidenced by cell shrinkage, detachment, and reduced density. In silico analysis, the molecular docked complexes showed the strong binding affinity of 8d (-11.01 kcal/mol) to Human 3α-HSD3, with key interactions involving TYR24, ASN56, and TRP227. Molecular dynamics simulations (100 ns) confirmed complex stability, with minimal RMSD/RMSF variations and sustained hydrogen bonding. Radius of gyration and SASA analyses suggested increased protein compactness upon binding. MM-GBSA binding free energy was 11.29 ± 7.31 kcal/mol, dominated by van der Waals contributions. These results highlighted compound 8d as a promising scaffold for further development as a breast cancer therapeutic targeting Human 3α-HSD3.