Synthesis, Evaluation of the Cytotoxicity, Apoptosis Induction in AGS Cell Line and Gene Expression and Molecular Modeling Studies of Novel Tetrahydropyrimidine Derivatives

Cancer is a complex disease that remains a leading cause of death worldwide owing to the lack of effective and efficient drugs. Compounds known as tetrahydropyrmidine have shown great potential as anti-cancer agents. Here, novel compounds based on tetrahydropyrimidine scaffold were successfully synthesized by the multicomponent reaction of Biginelli reaction. The biological functions of the selected compounds in AGS cells, including cytotoxicity, apoptosis induction, caspase3/7 activity, and gene expressions such as P53, CDNK2A, and Caspase-8/9, were evaluated. All synthesized compounds displayed moderate to good activity against the AGS cell line. Four compounds 4c, 4d, 4i, and 4 m induced apoptosis and activated caspase-3/7. Among them, compound 4c exhibited the highest cytotoxicity (IC50 = 69.60 µM), apoptosis induction, and caspase-3/7 activity. In comparison to other compounds, 4i showed a higher stimulation in the expression of P53, CDNK2A, and Caspase 9 genes. However, even though there was an increasing trend in the genes related to apoptosis and the cell cycle pathways, and the expression of caspase 9 was higher than caspase 8, these differences were not large enough to be significant. In addition, molecular docking studies and MD simulations were carried out to recognize the likely mechanism of the anti-cancer effect. Findings of docking revealed compounds fitted in the active site with good binding. MD simulations showed that compounds 4c, 4d and 4 m made a steady complex with the Eg5 enzyme. Thus, the possible mechanism of anti-cancer activity of these compounds can be through inhibition of the Eg5 enzyme.