Design, synthesis, bioactivity, X-ray crystallography, and molecular docking studies of chrysin-1,3,5-triazine...

In order to discover new effective anti-cancer drugs, fifteen derivatives of chrysin-1,3,5-triazine were designed and synthesized as potential novel anti-cancer agents. The structure of the target compounds were characterized by ¹H NMR, ¹³C NMR, IR, and HR-MS. The purity of all compounds were detected by HPLC. The structure of compound 4a was subjected to further investigation through single crystal X-ray diffraction, and elaborate discussions were conducted on the Hirshfeld surface and two-dimensional fingerprint diagram to explore the molecular conformation, crystal packing mode and molecular interactions. The antiproliferative activity of these compounds was assessed by the MTT (methylthiazolyl tetrazolium) assay against MDA-MB-231 (breast cancer cells), HeLa (cervical cancer cells), HCCLM3 (liver cancer cells), and HCT116 (colon cancer cells). Positive controls were cisplatin and chrysin. The results show that some chrysin-1,3,5-triazine derivatives have better anti-cancer activity than cisplatin and chrysin. Chrysin-1,3,5-triazine derivatives selectively targets HeLa cervical cancer cells. Compound 4c (7-((4-(Dibutylamino)-1,3,5-triazin-2-yl)oxy)-5-hydroxy-2-phenyl-4H-chromen-4-one) exhibits the strongest antiproliferative activity against HeLa cells (IC₅₀ = 9.86 ± 0.37 μM), superior to cisplatin and chrysin (IC₅₀ values of 28.09 ± 0.47 μM and 29.51 ± 0.51 μM, respectively). Further studies showed that compound 4c not only inhibits the invasion, adhesion, and proliferation of HeLa cells, but also has a strong inhibitory effect on the proliferation of HeLa tumor heterotopic xenografts in vivo. Molecular docking studies suggest that compound 4c can interact with phosphatidylinositol 3-kinase(PI3K)and cysteine aspartic acid proteinase-3 (Caspase-3). Western blot results demonstrated that compound 4c inhibited PI3K expression at the protein level and promoted the degradation of Pro-caspase-3, thereby activating the caspase-3-dependent apoptotic pathway. Furthermore, the absorption, distribution, metabolism, excretion, and toxicity (ADMET) properties of these compounds suggest that they may have pharmacological characteristics and safety. Thus, compound 4c has the potential to be a highly promising candidate for cancer treatment.