Design and Characterization of the First Selective and Potent Mechanism-Based Inhibitor of Cytochrome P450 4Z1

J Med Chem. 2020 May 14;63(9):4824-4836. doi: 10.1021/acs.jmedchem.0c00101. Epub 2020 Apr 30.


Mammary-tissue-restricted cytochrome P450 4Z1 (CYP4Z1) has garnered interest for its potential role in breast cancer progression. CYP4Z1-dependent metabolism of arachidonic acid preferentially generates 14,15-epoxyeicosatrienoic acid (14,15-EET), a metabolite known to influence cellular proliferation, migration, and angiogenesis. In this study, we developed time-dependent inhibitors of CYP4Z1 designed as fatty acid mimetics linked to the bioactivatable pharmacophore, 1-aminobenzotriazole (ABT). The most potent analogue, 8-[(1H-benzotriazol-1-yl)amino]octanoic acid (7), showed a 60-fold lower shifted-half-maximal inhibitory concentration (IC50) for CYP4Z1 compared to ABT, efficient mechanism-based inactivation of the enzyme evidenced by a KI = 2.2 μM and a kinact = 0.15 min-1, and a partition ratio of 14. Furthermore, 7 exhibited low off-target inhibition of other CYP isozymes. Finally, low micromolar concentrations of 7 inhibited 14,15-EET production in T47D breast cancer cells transfected with CYP4Z1. This first-generation, selective mechanism-based inhibitor (MBI) will be a useful molecular tool to probe the biochemical role of CYP4Z1 and its association with breast cancer.

MeSH terms

  • Arachidonic Acid / metabolism
  • Cell Line, Tumor
  • Cytochrome P-450 Enzyme Inhibitors / chemical synthesis
  • Cytochrome P-450 Enzyme Inhibitors / metabolism
  • Cytochrome P-450 Enzyme Inhibitors / pharmacology*
  • Cytochrome P450 Family 4 / antagonists & inhibitors*
  • Drug Design
  • Fatty Acids / chemical synthesis
  • Fatty Acids / metabolism
  • Fatty Acids / pharmacology*
  • Humans
  • Microsomes, Liver / metabolism
  • Oxidation-Reduction
  • Triazoles / chemical synthesis
  • Triazoles / metabolism
  • Triazoles / pharmacology*


  • Cytochrome P-450 Enzyme Inhibitors
  • Fatty Acids
  • Triazoles
  • Arachidonic Acid
  • CYP4Z1 protein, human
  • Cytochrome P450 Family 4