Synthesis, characterization, anticancer potential and mechanisms of cytotoxic activity of phosphoramide derivatives; experimental and theoretical study

Three new derivatives of phosphoramides with the formula of (C4H10O2) P(O) [C9H8N3] (P1), [(C4H10O2) P(O)]2 (C3H4N4S) (P2), and [(C6H5Cl) P(O)]2 (C2HN3S2) (P3) containing thiazole and pyrazole compounds were synthesized and characterized using techniques such as IR, 1H NMR, 13C NMR, 31P NMR, and Mass spectroscopies. The MTT assay was performed to examine the cytotoxicity of three compounds on A549 and HepG2 cell lines and determine the side effects of compounds, adipose-derived stem cells (ADSC) were used as the normal cells. The in-vitro results indicated a dose-dependent cytotoxic effect of the compounds in cancerous cells. However, the P2 has better anticancer performance without side effects on normal cells, and the P3 can be a good choice for anticancer activity. Moreover, Annexin V/PI flow cytometry results indicated that apoptosis, but not necrosis, is a significant type of cell death in treated cells in response to P2 and P3 compounds. Quantum calculations based on DFT-D were applied to obtain optimized structures and their energies, as well as HOMO and LUMO. Additionally, molecular docking studies were used to interact with receptors (DNA and DNA Topo I) to better understand the mechanisms of cytotoxic activity.