Pyrazolone functionalized heterocycles promote breast carcinoma cells ferroptosis via suppressing the Nrf-2/ HMOX-1/GPX-4 signaling axis: combined In Silico and In Vitro investigations.

Ferroptosis is a sort of cell death attributed to iron-dependent lipid peroxidation that represents a promising target for cancer therapy. Pyrazolone candidates have a broad pharmacological potential, including antineoplastic activity. In this study, a series of pyrazolone-ornamented heterocycles (3-10) was synthesized and characterized via elemental analysis and different spectroscopic data, including NMR, FT-IR, and mass spectroscopy. Molecular docking and SwissADME analyses identified compound 10 as exhibiting the highest binding affinity toward the ferroptosis-related targets nuclear factor erythroid 2-related factor 2 (Nrf-2) and glutathione peroxidase-4 (GPX-4) compared with the reference drug cisplatin, alongside favorable pharmacokinetic and drug-likeness characteristics. DFT/B3LYP/6-311+G (d,p) calculations were carried out on the most active compound to gain insight into its optimized molecular structure and electronic properties. The electronic properties were evaluated through frontier molecular orbital analysis and MEP map. Also, ELF, LOL, and NCI analyses were performed to elucidate the degree of electron localization and to examine the type and intensity of interactions. within the studied system. In vitro, compound 10 exerted selective cytotoxicity against MCF-7 neoplastic cells with pronounced morphological alterations. Flow cytometry revealed that compound 10 elevated intracellular reactive oxygen species (ROS) levels, enhancing oxidative stress, which was substantiated by raised malondialdehyde (MDA) levels and a drop in reduced glutathione (GSH) content and GPX activity. It also raised iron levels, suggesting iron-catalyzed lipid peroxidation. Mechanistically, compound 10 downregulated Nrf-2 protein expression, reduced heme oxygenase-1 (HMOX-1) and elevated p53 gene expression in a dose-dependent manner. Collectively, compound 10 contributed to cancer cell death via inducing oxidative stress-mediated ferroptosis; hence, compound 10 may be a promising treatment for breast carcinoma.