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Chemical synthesis of a reported p47phox/p22phox inhibitor and characterization of its instability and irreproducible activity

The nicotinamide adenine dinucleotide phosphate oxidase 2 (NOX2) multi-subunit complex is a highly abundant and central source of reactive oxygen species. NOX2 is a key enzyme of the innate immune system involved in antibacterial response, but excessive NOX2 activity is involved in oxidative stress...

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Autores principales: Zang, Jie, Cambet, Yves, Jaquet, Vincent, Bach, Anders
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9850094/
https://www.ncbi.nlm.nih.gov/pubmed/36686674
http://dx.doi.org/10.3389/fphar.2022.1075328
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author Zang, Jie
Cambet, Yves
Jaquet, Vincent
Bach, Anders
author_facet Zang, Jie
Cambet, Yves
Jaquet, Vincent
Bach, Anders
author_sort Zang, Jie
collection PubMed
description The nicotinamide adenine dinucleotide phosphate oxidase 2 (NOX2) multi-subunit complex is a highly abundant and central source of reactive oxygen species. NOX2 is a key enzyme of the innate immune system involved in antibacterial response, but excessive NOX2 activity is involved in oxidative stress and inflammation in many diseases. Inhibition of NOX2 has great potential as a therapeutic strategy. An intriguing pharmacological approach for inhibiting NOX2 is to target the p47phox subunit and thereby block the protein-protein interaction with p22phox, whereby assembling and activation of NOX2 is prevented. However, the shallow binding pocket of p47phox makes it difficult to develop drug-like p47phox/p22phox inhibitors. Recently, the small molecule LMH001 was reported to inhibit the p47phox/p22phox interaction, reduce endothelial NOX2 activity, and protect mice from angiotensin II-induced vascular oxidative stress. These noteworthy results could have significant impact on the field of NOX2 pharmacology, as specific and efficient inhibitors are scarce. Here, we synthesized and tested LMH001 to have it available as a positive control. We established a robust synthetic route for providing LMH001, but subsequently we experienced that LMH001 is chemically unstable in aqueous buffer. In addition, neither LMH001 nor its breakdown products were able to inhibit the p47phox/p22phox interaction in a non-cellular fluorescence polarization assay. However, LHM001 was a weak inhibitor of NOX2 in a functional cell assay, but with same low potency as one of its breakdown products. These findings question the activity and suggested mechanism of LMH001 and constitute important information for other researchers interested in chemical probes for studying NOX2 biology.
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spelling pubmed-98500942023-01-20 Chemical synthesis of a reported p47phox/p22phox inhibitor and characterization of its instability and irreproducible activity Zang, Jie Cambet, Yves Jaquet, Vincent Bach, Anders Front Pharmacol Pharmacology The nicotinamide adenine dinucleotide phosphate oxidase 2 (NOX2) multi-subunit complex is a highly abundant and central source of reactive oxygen species. NOX2 is a key enzyme of the innate immune system involved in antibacterial response, but excessive NOX2 activity is involved in oxidative stress and inflammation in many diseases. Inhibition of NOX2 has great potential as a therapeutic strategy. An intriguing pharmacological approach for inhibiting NOX2 is to target the p47phox subunit and thereby block the protein-protein interaction with p22phox, whereby assembling and activation of NOX2 is prevented. However, the shallow binding pocket of p47phox makes it difficult to develop drug-like p47phox/p22phox inhibitors. Recently, the small molecule LMH001 was reported to inhibit the p47phox/p22phox interaction, reduce endothelial NOX2 activity, and protect mice from angiotensin II-induced vascular oxidative stress. These noteworthy results could have significant impact on the field of NOX2 pharmacology, as specific and efficient inhibitors are scarce. Here, we synthesized and tested LMH001 to have it available as a positive control. We established a robust synthetic route for providing LMH001, but subsequently we experienced that LMH001 is chemically unstable in aqueous buffer. In addition, neither LMH001 nor its breakdown products were able to inhibit the p47phox/p22phox interaction in a non-cellular fluorescence polarization assay. However, LHM001 was a weak inhibitor of NOX2 in a functional cell assay, but with same low potency as one of its breakdown products. These findings question the activity and suggested mechanism of LMH001 and constitute important information for other researchers interested in chemical probes for studying NOX2 biology. Frontiers Media S.A. 2023-01-05 /pmc/articles/PMC9850094/ /pubmed/36686674 http://dx.doi.org/10.3389/fphar.2022.1075328 Text en Copyright © 2023 Zang, Cambet, Jaquet and Bach. 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 Pharmacology
Zang, Jie
Cambet, Yves
Jaquet, Vincent
Bach, Anders
Chemical synthesis of a reported p47phox/p22phox inhibitor and characterization of its instability and irreproducible activity
title Chemical synthesis of a reported p47phox/p22phox inhibitor and characterization of its instability and irreproducible activity
title_full Chemical synthesis of a reported p47phox/p22phox inhibitor and characterization of its instability and irreproducible activity
title_fullStr Chemical synthesis of a reported p47phox/p22phox inhibitor and characterization of its instability and irreproducible activity
title_full_unstemmed Chemical synthesis of a reported p47phox/p22phox inhibitor and characterization of its instability and irreproducible activity
title_short Chemical synthesis of a reported p47phox/p22phox inhibitor and characterization of its instability and irreproducible activity
title_sort chemical synthesis of a reported p47phox/p22phox inhibitor and characterization of its instability and irreproducible activity
topic Pharmacology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9850094/
https://www.ncbi.nlm.nih.gov/pubmed/36686674
http://dx.doi.org/10.3389/fphar.2022.1075328
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