NK-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 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.
Authors
- Céleste Sèle (ORCID: https://orcid.org/0000-0003-4916-3065)
- Wolfgang Knecht (ORCID: https://orcid.org/0000-0001-9530-7598)
- Suraj Verma (ORCID: https://orcid.org/0000-0002-9684-0885)
- Ewa Krupinska
- Claudio Angione (ORCID: https://orcid.org/0000-0002-3140-7909)
- Suresh Sagadevan (ORCID: https://orcid.org/0000-0003-2779-6339)
- Javier Ramón (ORCID: https://orcid.org/0000-0002-0673-9338)
- Pablo Huertas (ORCID: https://orcid.org/0000-0002-1756-4449)
- Néstor García‐Rodríguez (ORCID: https://orcid.org/0000-0002-4049-1604)
- M. Angeles Juanes (ORCID: https://orcid.org/0000-0002-9801-9652)
- Ramón Martí (ORCID: https://orcid.org/0000-0002-8273-9540)
- Salvador Meseguer (ORCID: https://orcid.org/0000-0002-2718-4351)
- Paloma Ordóñez‐Morán (ORCID: https://orcid.org/0000-0001-7688-1252)
- Annalisa Occhipinti (ORCID: https://orcid.org/0000-0001-6075-1496)
- Mohammed Moustapha Anwar (ORCID: https://orcid.org/0000-0001-5035-8463)
- Aida Rodríguez-Jiménez
Institutions
- Universitat Autònoma de Barcelona (ES)
- University of Nottingham (GB)
- Lund University (SE)
- Vall d'Hebron Institut de Recerca (ES)
- Centre for Biomedical Network Research on Rare Diseases (ES)
- Centro Andaluz de Biología Molecular y Medicina Regenerativa (ES)
- Centro de Investigacion Principe Felipe (ES)
- Teesside University (GB)
Publication Details
- Journal
- Journal of Experimental & Clinical Cancer Research
- Published
- 2025-10-17
- DOI
- https://doi.org/10.1186/s13046-025-03538-w
- Primary Topic
- Biochemical and Molecular Research
- Type
- article
- Field-Weighted Citation Impact
- 0.00
Funders
- Fundació la Marató de TV3
- Universidad Complutense de Madrid
- Lunds Universitet
- Generalitat Valenciana
- Junta de Andalucía
- Instituto de Salud Carlos III
- European Regional Development Fund
- Agencia Estatal de Investigación