Development of novel coumarin-barbituric acid hybrids as a new class of anti-breast cancer agents.

Inspired by the known anti-breast cancer activity of barbituric acid derivatives and the anti-tubulin properties of the coumarin scaffold, a novel series of triazole-linked coumarin-barbituric acid hybrids was designed by a molecular hybridization approach, synthesized, and evaluated for their potential efficacy against breast cancer. Among the synthesized compounds, PKA2 emerged as the most potent against MCF-7 breast cancer cells, exhibiting an IC50 of 1.734 μM. PKA2 also demonstrated significant inhibition of tubulin polymerization (IC50 = 3.216 μM) and triggered apoptosis in cancer cells by inducing G2/M phase cell cycle arrest. Notably, PKA2 showed a high degree of selectivity toward MCF-7 cells over normal skin fibroblast (L929) cells, with a selectivity index of 10.123, suggesting preferential cytotoxicity toward cancer cells. Morphological analyses confirmed that PKA2 induces cell death primarily through apoptotic mechanisms. Additionally, molecular docking and molecular dynamics simulations supported the interaction of PKA2 at the colchicine-binding site of tubulin, aligning with its observed mechanism of action. Collectively, these findings highlight PKA2 as a promising lead compound for the development of selective and potent anti-breast cancer agents targeting microtubule dynamics.