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

Umair Z. Malik, Nicholas J. Hundley, Guillermo Romero, Rafael Radi, Bruce A. Freeman, Margaret M. Tarpey, Eric E. Kelley

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109 Scopus citations


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 (IC 50 = 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 (IC 50 = 4.4 nM) while further limiting the effect of allopurinol (IC 50 = 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.

Original languageEnglish (US)
Pages (from-to)179-184
Number of pages6
JournalFree Radical Biology and Medicine
Issue number1
StatePublished - Jul 1 2011

Bibliographical note

Funding Information:
This work was supported by AHA Scientist Development Grant 10SDG3560005 and a University of Pittsburgh Vascular Medicine Institute Seed Grant (E.E.K.), Comisión Sectorial de Investigación Científica, Universidad de la República (Uruguay), Agencia Nacional de Investigación e Innovación (FCE2009_2486), Uruguay, and the Howard Hughes Medical Institute (R.R.), and NIH HL8115 and HL64937 (B.A.F.).


  • Allopurinol
  • Endothelial cells
  • Febuxostat
  • Free radicals
  • Inflammation
  • Oxypurinol
  • Reactive oxygen species
  • Uloric
  • Xanthine oxidase
  • Xanthine oxidase inhibitors


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