In vitro and in silico evaluation of the anti-cancer effects of phenyramine hydrogen maleat on a lung cancer model within the scope of drug repositioning

Objective: The aim of this thesis is to unveil the anti-cancer potential of Pheniramine Hydrogen Maleate (PHM), an H1 receptor antagonist commonly used in allergy treatment, through in silico and in vitro experiments. This approach utilizes drug repositioning, a method known to significantly reduce the costs and time required for the development of new drugs. Methods: Similarities between PHM and other therapeutic drugs were analyzed using the DrugCentral 2023 web server. The interaction with the FasL receptor, which is involved in the extrinsic apoptotic pathway, was examined in silico through ligand/protein molecular docking (using BIOVIA Discovery Studio Visualizer). The cytotoxic effects of PHM were investigated in vitro using A549 human lung cancer cells. Results: According to the L1000 gene profile similarity analysis on the DrugCentral 2023 web server, PHM showed similarities to a total of 13,106 drugs, with 213 of them being known for their anti-cancer effects, exhibiting a Root Mean Square Deviation (RMSD) value below 1. Molecular docking analysis indicated that PHM has the potential to bind to the third active site of the receptor protein, thereby altering its three-dimensional conformation and initiating extrinsic apoptotic signaling. The effects of PHM on cell viability were assessed using the MTT assay, revealing that it inhibits colony formation and cell migration. According to our ELISA results, caspase-7 levels increased at 100 and 50 µg/ml doses at the 24-hour mark, but this increase could not be confirmed by western blot analysis. However, at 48 hours, an increase in caspase-7 was consistent with western blot findings, indicating apoptosis-induced cell death in A549 cells. Conclusion: PHM exhibits structural similarity to 213 drugs used in cancer treatment. Molecular docking analysis suggests that it has the potential to initiate extrinsic apoptotic signaling. It demonstrates cytotoxic effects on A549 cells, and apoptosis is confirmed to be mediated via caspase-7 through ELISA and western blot methods.