Febuxostat inhibition of endothelial-bound XO: implications for targeting vascular ROS production

Free Radic Biol Med. 2011 Jul 1;51(1):179-84. doi: 10.1016/j.freeradbiomed.2011.04.004. Epub 2011 Apr 15.


Xanthine oxidase (XO) is a critical source of reactive oxygen species (ROS) that contribute to vascular inflammation. Binding of XO to vascular endothelial cell glycosaminoglycans (GAGs) results in significant resistance to inhibition by traditional pyrazolopyrimidine-based inhibitors such as allopurinol. Therefore, we compared the extent of XO inhibition (free and GAG-bound) by allopurinol to that by febuxostat, a newly approved nonpurine XO-specific inhibitor. In solution, febuxostat was 1000-fold more potent than allopurinol at inhibiting XO-dependent uric acid formation (IC₅₀= 1.8 nM vs 2.9 μM). Association of XO with heparin-Sepharose 6B (HS6B-XO) had minimal effect on the inhibition of uric acid formation by febuxostat (IC₅₀= 4.4 nM) while further limiting the effect of allopurinol (IC₅₀= 64 μM). Kinetic analysis of febuxostat inhibition revealed K(i) values of 0.96 (free) and 0.92 nM (HS6B-XO), confirming equivalent inhibition for both free and GAG-immobilized enzyme. When XO was bound to endothelial cell GAGs, complete enzyme inhibition was observed with 25 nM febuxostat, whereas no more than 80% inhibition was seen with either allopurinol or oxypurinol, even at concentrations above those tolerated clinically. The superior potency for inhibition of endothelium-associated XO is predictive of a significant role for febuxostat in investigating pathological states in which XO-derived ROS are contributive and traditional XO inhibitors are only slightly effective.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Allopurinol / pharmacology
  • Animals
  • Cattle
  • Cells, Cultured
  • Endothelial Cells / metabolism*
  • Enzyme Inhibitors / pharmacology*
  • Febuxostat
  • Glycosaminoglycans / metabolism
  • Kinetics
  • Oxypurinol / pharmacology
  • Reactive Oxygen Species / metabolism*
  • Thiazoles / pharmacology*
  • Xanthine Oxidase / antagonists & inhibitors*
  • Xanthine Oxidase / metabolism


  • Enzyme Inhibitors
  • Glycosaminoglycans
  • Reactive Oxygen Species
  • Thiazoles
  • Febuxostat
  • Allopurinol
  • Xanthine Oxidase
  • Oxypurinol