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Novel Starting Points for Human Glycolate Oxidase Inhibitors, Revealed by Crystallography-Based Fragment Screening

Primary hyperoxaluria type I (PH1) is caused by AGXT gene mutations that decrease the functional activity of alanine:glyoxylate aminotransferase. A build-up of the enzyme’s substrate, glyoxylate, results in excessive deposition of calcium oxalate crystals in the renal tract, leading to debilitating...

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Autores principales: Mackinnon, Sabrina R., Bezerra, Gustavo A., Krojer, Tobias, Szommer, Tamas, von Delft, Frank, Brennan, Paul E., Yue, Wyatt W.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9114433/
https://www.ncbi.nlm.nih.gov/pubmed/35601556
http://dx.doi.org/10.3389/fchem.2022.844598
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author Mackinnon, Sabrina R.
Bezerra, Gustavo A.
Krojer, Tobias
Szommer, Tamas
von Delft, Frank
Brennan, Paul E.
Yue, Wyatt W.
author_facet Mackinnon, Sabrina R.
Bezerra, Gustavo A.
Krojer, Tobias
Szommer, Tamas
von Delft, Frank
Brennan, Paul E.
Yue, Wyatt W.
author_sort Mackinnon, Sabrina R.
collection PubMed
description Primary hyperoxaluria type I (PH1) is caused by AGXT gene mutations that decrease the functional activity of alanine:glyoxylate aminotransferase. A build-up of the enzyme’s substrate, glyoxylate, results in excessive deposition of calcium oxalate crystals in the renal tract, leading to debilitating renal failure. Oxidation of glycolate by glycolate oxidase (or hydroxy acid oxidase 1, HAO1) is a major cellular source of glyoxylate, and siRNA studies have shown phenotypic rescue of PH1 by the knockdown of HAO1, representing a promising inhibitor target. Here, we report the discovery and optimization of six low-molecular-weight fragments, identified by crystallography-based fragment screening, that bind to two different sites on the HAO1 structure: at the active site and an allosteric pocket above the active site. The active site fragments expand known scaffolds for substrate-mimetic inhibitors to include more chemically attractive molecules. The allosteric fragments represent the first report of non-orthosteric inhibition of any hydroxy acid oxidase and hold significant promise for improving inhibitor selectivity. The fragment hits were verified to bind and inhibit HAO1 in solution by fluorescence-based activity assay and surface plasmon resonance. Further optimization cycle by crystallography and biophysical assays have generated two hit compounds of micromolar (44 and 158 µM) potency that do not compete with the substrate and provide attractive starting points for the development of potent and selective HAO1 inhibitors.
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spelling pubmed-91144332022-05-19 Novel Starting Points for Human Glycolate Oxidase Inhibitors, Revealed by Crystallography-Based Fragment Screening Mackinnon, Sabrina R. Bezerra, Gustavo A. Krojer, Tobias Szommer, Tamas von Delft, Frank Brennan, Paul E. Yue, Wyatt W. Front Chem Chemistry Primary hyperoxaluria type I (PH1) is caused by AGXT gene mutations that decrease the functional activity of alanine:glyoxylate aminotransferase. A build-up of the enzyme’s substrate, glyoxylate, results in excessive deposition of calcium oxalate crystals in the renal tract, leading to debilitating renal failure. Oxidation of glycolate by glycolate oxidase (or hydroxy acid oxidase 1, HAO1) is a major cellular source of glyoxylate, and siRNA studies have shown phenotypic rescue of PH1 by the knockdown of HAO1, representing a promising inhibitor target. Here, we report the discovery and optimization of six low-molecular-weight fragments, identified by crystallography-based fragment screening, that bind to two different sites on the HAO1 structure: at the active site and an allosteric pocket above the active site. The active site fragments expand known scaffolds for substrate-mimetic inhibitors to include more chemically attractive molecules. The allosteric fragments represent the first report of non-orthosteric inhibition of any hydroxy acid oxidase and hold significant promise for improving inhibitor selectivity. The fragment hits were verified to bind and inhibit HAO1 in solution by fluorescence-based activity assay and surface plasmon resonance. Further optimization cycle by crystallography and biophysical assays have generated two hit compounds of micromolar (44 and 158 µM) potency that do not compete with the substrate and provide attractive starting points for the development of potent and selective HAO1 inhibitors. Frontiers Media S.A. 2022-05-04 /pmc/articles/PMC9114433/ /pubmed/35601556 http://dx.doi.org/10.3389/fchem.2022.844598 Text en Copyright © 2022 Mackinnon, Bezerra, Krojer, Szommer, von Delft, Brennan and Yue. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Chemistry
Mackinnon, Sabrina R.
Bezerra, Gustavo A.
Krojer, Tobias
Szommer, Tamas
von Delft, Frank
Brennan, Paul E.
Yue, Wyatt W.
Novel Starting Points for Human Glycolate Oxidase Inhibitors, Revealed by Crystallography-Based Fragment Screening
title Novel Starting Points for Human Glycolate Oxidase Inhibitors, Revealed by Crystallography-Based Fragment Screening
title_full Novel Starting Points for Human Glycolate Oxidase Inhibitors, Revealed by Crystallography-Based Fragment Screening
title_fullStr Novel Starting Points for Human Glycolate Oxidase Inhibitors, Revealed by Crystallography-Based Fragment Screening
title_full_unstemmed Novel Starting Points for Human Glycolate Oxidase Inhibitors, Revealed by Crystallography-Based Fragment Screening
title_short Novel Starting Points for Human Glycolate Oxidase Inhibitors, Revealed by Crystallography-Based Fragment Screening
title_sort novel starting points for human glycolate oxidase inhibitors, revealed by crystallography-based fragment screening
topic Chemistry
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9114433/
https://www.ncbi.nlm.nih.gov/pubmed/35601556
http://dx.doi.org/10.3389/fchem.2022.844598
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