Discovery of pyranocoumarin derivatives as dual cytotoxic activity against the resistant cancer cell MCF-7/ADR and inhibitory effect of the P-glycoprotein expression levels and function

The creation of new molecules that enhance biological activity and target as dual cytotoxic activity against the resistant cancer cell MCF-7/ADR and inhibitory effect of the P-glycoprotein expression levels and function is essential for the progression of cancer treatment. This study synthesized and evaluated a series of halogenated pyranocoumarin derivatives (4a-d) for their potential as anti-cancer agents, targeting the reduction of P-glycoprotein (P-gp) overexpression and drug efflux. The target compounds fully confirm the chemical structures via IR, MS, 1D/2D NMR and evaluated for their antiproliferative activity against three human cancer cell lines, MCF-7, MCF-7/ADR, Caco-2 and two normal cell lines, HFL-1 and WI-38. Compounds 4a-d were assigned to be the most potent derivatives against MCF-7, MCF-7/ADR, and Caco-2 cell lines in comparison with Sorafenib and Doxorubicin and exhibited higher IC₅₀ values in normal cells compared to cancer cells. Several compounds 4a,b,d possessed a relatively good potency against MCF-7/ADR cells as compared to the standard drugs and were tested as a P-gp inhibitor. Moreover, the halogenated substituents (4a,b,d), 4-F, 4-Cl, and 4-I; have good potency against P-gp mediated MDR in MCF-7/ADR in comparison to Doxorubicin. Meanwhile, Rho123 accumulation assays revealed that only compound 4d effectively inhibited P-pg and efflux function. The SAR of 4a-d demonstrate promising anticancer activity, with the iodo-substituted (4d) showing superior potency across all cancer cell lines, and P-gp inhibitory activity in MCF-7/ADR cells. Molecular docking analysis shows that 4d emerges as the most promising candidate based on its superior binding affinity, interaction energy. The ADMET analysis show that compounds 4a-d has promising physicochemical properties and ADMET profiles. Of these, 4d stands out as the most promising candidate. Additionally, molecular dynamics (MD) simulations are employed to clarify the dynamic properties and structural flexibility of P-glycoprotein with most active 4d, which highlighted the stability and rigidity of distinct protein regions, particularly the active site.