Discovery of piperine derivatives as inhibitors of human dihydroorotate dehydrogenase to induce ferroptosis in cancer cells.

[Display omitted] • Piperine derivatives were designed and synthesized as novel h DHODH inhibitors. • H19 showed the strongest cytotoxic and h DHODH inhibitory activities among all the synthesized compounds. • H19 could effectively inhibit the proliferation of NCI-H226 cells by inducing ferroptosis. Inhibition of human dihydroorotate dehydrogenase (h DHODH) represents a promising strategy for suppressing the proliferation of cancer cells. To identify novel and potent h DHODH inhibitors, a total of 28 piperine derivatives were designed and synthesized. Their cytotoxicities against three human cancer cell lines (NCI-H226, HCT-116, and MDA-MB-231) and h DHODH inhibitory activities were also evaluated. Among them, compound H19 , exhibited the strongest inhibitory activities (NCI-H226 IC 50 = 0.95 µM, h DHODH IC 50 = 0.21 µM). Further pharmacological investigations revealed that H19 exerted anticancer effects by inducing ferroptosis in NCI-H226 cells, with its cytotoxicity being reversed by ferroptosis inhibitors. This was supported by the intracellular growth or decline of ferroptosis markers, including lipid peroxidation, Fe2+, GSH, and 4-HNE. Overall, H19 emerges as a promising h DHODH inhibitor with potential anticancer properties warranting development. [ABSTRACT FROM AUTHOR]