Bifunctional Naphtho[2,3- d][1,2,3]triazole-4,9-dione Compounds Exhibit Antitumor Effects In Vitro and In Vivo by Inhibiting Dihydroorotate Dehydrogenase and Inducing Reactive Oxygen Species Production

J Med Chem. 2020 Jul 23;63(14):7633-7652. doi: 10.1021/acs.jmedchem.0c00512. Epub 2020 Jul 1.


Human dihydroorotate dehydrogenase (hDHODH) is an attractive target for cancer therapy. Based on its crystal structure, we designed and synthesized a focused compound library containing the structural moiety of 1,4-benzoquinone, which possesses reactive oxygen species (ROS) induction capacity. Compound 3s with a naphtho[2,3-d][1,2,3]triazole-4,9-dione scaffold exhibited inhibitory activity against hDHODH. Further optimization led to compounds 11k and 11l, which inhibited hDHODH activity with IC50 values of 9 and 4.5 nM, respectively. Protein-ligand cocrystal structures clearly depicted hydrogen bond and hydrophobic interactions of 11k and 11l with hDHODH. Compounds 11k and 11l significantly inhibited leukemia cell and solid tumor cell proliferation and induced ROS production, mitochondrial dysfunction, apoptosis, and cell cycle arrest. Nanocrystallization of compound 11l displayed significant in vivo antitumor effects in the Raji xenograft model. Overall, this study provides a novel bifunctional compound 11l with hDHODH inhibition and ROS induction efficacy, which represents a promising anticancer lead worthy of further exploration.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antineoplastic Agents / chemical synthesis
  • Antineoplastic Agents / metabolism
  • Antineoplastic Agents / pharmacokinetics
  • Antineoplastic Agents / pharmacology*
  • Apoptosis / drug effects
  • Cell Line, Tumor
  • Cell Proliferation / drug effects
  • Dihydroorotate Dehydrogenase
  • Drug Screening Assays, Antitumor
  • Enzyme Inhibitors / chemical synthesis
  • Enzyme Inhibitors / metabolism
  • Enzyme Inhibitors / pharmacokinetics
  • Enzyme Inhibitors / pharmacology*
  • Female
  • Humans
  • Male
  • Membrane Potential, Mitochondrial / drug effects
  • Mice, Inbred BALB C
  • Mice, SCID
  • Molecular Docking Simulation
  • Molecular Structure
  • Naphthoquinones / chemical synthesis
  • Naphthoquinones / metabolism
  • Naphthoquinones / pharmacokinetics
  • Naphthoquinones / pharmacology*
  • Oxidoreductases Acting on CH-CH Group Donors / antagonists & inhibitors*
  • Oxidoreductases Acting on CH-CH Group Donors / metabolism
  • Protein Binding
  • Rats, Sprague-Dawley
  • Reactive Oxygen Species / metabolism*
  • S Phase Cell Cycle Checkpoints / drug effects
  • Structure-Activity Relationship
  • Triazoles / chemical synthesis
  • Triazoles / metabolism
  • Triazoles / pharmacokinetics
  • Triazoles / pharmacology*


  • Antineoplastic Agents
  • Dihydroorotate Dehydrogenase
  • Enzyme Inhibitors
  • Naphthoquinones
  • Reactive Oxygen Species
  • Triazoles
  • Oxidoreductases Acting on CH-CH Group Donors