biostudies-literatureAnwar MMMICIU/AEI/10.13039/501100011033 and by ERDF/EUEMERGIA 2021 program from the Andalusian Regional Government-Junta de AndalucíaMICIU/AEI/10.13039/501100011033MCIN/AEI/10.13039/501100011033CIDEGENT Excellent Research Program of the Valencian regional governmentInstituto de Salud Carlos III cofunded with ERDFFundació la Marató de TV3292https://www.ebi.ac.uk/biostudies/studies/S-EPMC12534982biostudies-literatureUnknown44(1)Human dihydroorotate dehydrogenase (DHODH) is the rate-limiting enzyme in pyrimidine de novo synthesis and represents a promising target for cancer therapy. However, current inhibitors of DHODH have limited clinical effectiveness and adverse effects. Herein, we report NK-A 17E-233I, a novel small-molecule inhibitor of the human DHODH enzyme, identified through a prospective virtual screening methodology. Molecular docking and biochemical assays show NK-A 17E-233I functions as a pure or partial competitive inhibitor with respect to the natural substrate, dihydroorotate (DHO). It adopts a distinct binding mode from classical inhibitors that target the flavin mononucleotide (FMN) binding cavity of the hydrophobic tunnel. NK-A 17E-233I exhibits selective cytotoxicity in both human cancer cell Journal of experimental & clinical cancer research : CRNK-A 17E-233I: a novel competitive inhibitor of human dihydroorotate dehydrogenase (DHODH) for cancer therapy.PMC12534982PID2022-140899NB-I00PID2022-136791NB-I00CNS2024-154649CIDEGENT/2021/026EMC21_00057PI21/00554202020-30-31Garcia-Rodriguez NVerma SSagadevan SAngione COcchipinti AMarti RKnecht WOrdonez-Moran PJuanes MASele CHuertas PMeseguer SKrupinska EAnwar MMRamon JRodriguez-Jimenez AfalseNK-A 17E-233I: a novel competitive inhibitor of human dihydroorotate dehydrogenase (DHODH) for cancer therapy.Human dihydroorotate dehydrogenase (DHODH) is the rate-limiting enzyme in pyrimidine de novo synthesis and represents a promising target for cancer therapy. However, current inhibitors of DHODH have limited clinical effectiveness and adverse effects. Herein, we report NK-A 17E-233I, a novel small-molecule inhibitor of the human DHODH enzyme, identified through a prospective virtual screening methodology. Molecular docking and biochemical assays show NK-A 17E-233I functions as a pure or partial competitive inhibitor with respect to the natural substrate, dihydroorotate (DHO). It adopts a distinct binding mode from classical inhibitors that target the flavin mononucleotide (FMN) binding cavity of the hydrophobic tunnel. NK-A 17E-233I exhibits selective cytotoxicity in both human cancer cell 2025-01-01T00:00:00Z2025 Oct2026-05-09T03:21:35.079Z2026-05-09T03:11:54.469ZS-EPMC125349824110798510.1186/s13046-025-03538-w