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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

Characterization and screening of tight binding inhibitors of xanthine oxidase: an on-flow assay

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Author(s):
Rodrigues, M. V. N. [1, 2] ; Correa, R. S. [1] ; Vanzolini, K. L. [1] ; Santos, D. S. [3] ; Batista, A. A. [1] ; Cass, Q. B. [1]
Total Authors: 6
Affiliation:
[1] Univ Fed Sao Carlos, Dept Quim, BR-13565905 Sao Carlos, SP - Brazil
[2] Univ Estadual Campinas, Ctr Pluridisciplinar Pesquisas Quim Biol & Agr, BR-13140000 Paulinia, SP - Brazil
[3] Pontificia Univ Catolica Rio Grande do Sul, Ctr Pesquisas Biol Mol & Func, Inst Nacl Ciencia & Tecnol TB, BR-90619900 Porto Alegre, RS - Brazil
Total Affiliations: 3
Document type: Journal article
Source: RSC ADVANCES; v. 5, n. 47, p. 37533-37538, 2015.
Web of Science Citations: 6
Abstract

Xanthine oxidase (XO) is an enzyme in the purine salvage pathway that catalyzes the oxidation of hypoxanthine to xanthine with subsequent production of uric acid from the xanthine oxidation, and it has been considered an important target of newly developed inhibitors. Based on the advantages of using immobilized capillary enzyme reactors (ICERs) in a 2D LC system as a tool for screening new enzymatic ligands, this work validated an XO-ICER using allopurinol as a positive control. Despite the complex interaction between XO and allopurinol due its tight binding nature, it was possible to recognize the inhibitory kinetics parameters through Morrison's equation. The tight binding nature of inhibition was established by varying the IC50 values according to the substrate concentration. The kinetic inhibitory profile of allopurinol was used to validate the XO-ICER. Then, the XO-ICER was used to screen specific ruthenium derivatives. The selected compound, 4CBALO, an allopurinol ruthenium derivative, exhibited 100% inhibition at 200 mu M compared to 86% inhibition from allopurinol at the same concentration. The inhibitory effect on the immobilized XO was reversible after the elution of the compound, with immediate recovery of the ICER activity. Additionally, 4CBALO behaved as a selective and competitive tight binder of xanthine oxidase with a true K-i value of 0.29 mu M, which was obtained from the Morrison equation. This report describes the first on-flow characterization of tight binders of xanthine oxidase. (AU)

FAPESP's process: 13/01710-1 - Enzyme ligand: new models of screening
Grantee:Quezia Bezerra Cass
Support Opportunities: Research Projects - Thematic Grants
FAPESP's process: 09/08131-1 - Ruthenium complexes containing ligands with biologic interest: chemical and structural aspects and evaluation of their citotoxicity against tumoral cells
Grantee:Rodrigo de Souza Corrêa
Support Opportunities: Scholarships in Brazil - Doctorate
FAPESP's process: 13/26559-4 - Structural modifications in biologically active Ru(II) complexes toward the design of new metallodrug candidates
Grantee:Rodrigo de Souza Corrêa
Support Opportunities: Scholarships in Brazil - Post-Doctoral