NK-A 17E-233I: a novel competitive inhibitor of human dihydroorotate dehydrogenase (DHODH) for cancer therapy.

Anwar MM, Meseguer S, García-Rodríguez N, Krupinska E, Sele C, Rodríguez-Jiménez A, Verma S, Sagadevan S, Ramon J, Martí R, Occhipinti A, Angione C, Ordóñez-Morán P, Knecht W, Huertas P, Juanes MA

J. Exp. Clin. Cancer Res. 44 (1) 292 [2025-10-17; online 2025-10-17]

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 lines and patient-derived intestinal organoids, inducing DNA damage, S-phase arrest, and cell death. Unlike Brequinar, NK-A 17E-233I preserves mitochondrial respiration via complexes I and II and maintains ATP-linked basal respiration, avoiding the impairment of the electron transport chain (ETC). Our findings imply the aptitude of NK-A 17E-233I as a novel competitive inhibitor of human DHODH, representing a significant advancement in this field since the 1990s.

PubMed 41107985

DOI 10.1186/s13046-025-03538-w

Crossref 10.1186/s13046-025-03538-w

pmc: PMC12534982
pii: 10.1186/s13046-025-03538-w