Developing Inhibitors of the p47phox-p22phox Protein-Protein Interaction by Fragment-Based Drug Discovery

Sara Marie Øie Solbak, Jie Zang, Dilip Narayanan, Lars Jakobsen Høj, Saskia Bucciarelli, Charlotte Softley, Sebastian Meier, Annette Eva Langkilde, Charlotte Held Gotfredsen, Michael Sattler, Anders Bach*

*Corresponding author for this work

Research output: Contribution to journalJournal articleResearchpeer-review

Abstract

Nicotinamide adenine dinucleotide phosphate oxidase isoform 2 is an enzyme complex, which generates reactive oxygen species and contributes to oxidative stress. The p47phox-p22phox interaction is critical for the activation of the catalytical NOX2 domain, and p47phox is a potential target for therapeutic intervention. By screening 2500 fragments using fluorescence polarization and a thermal shift assay and validation by surface plasmon resonance, we found eight hits toward the tandem SH3 domain of p47phox (p47phoxSH3A-B) with KD values of 400-600 μM. Structural studies revealed that fragments 1 and 2 bound two separate binding sites in the elongated conformation of p47phoxSH3A-B and these competed with p22phox for binding to p47phoxSH3A-B. Chemical optimization led to a dimeric compound with the ability to potently inhibit the p47phoxSH3A-B-p22phox interaction (Ki of 20 μM). Thereby, we reveal a new way of targeting p47phox and present the first report of drug-like molecules with the ability to bind p47phox and inhibit its interaction with p22phox.
Original languageEnglish
JournalJournal of Medicinal Chemistry
Volume63
Issue number3
Pages (from-to)1156-1177
ISSN0022-2623
DOIs
Publication statusPublished - 2020

Cite this

Solbak, S. M. Ø., Zang, J., Narayanan, D., Høj, L. J., Bucciarelli, S., Softley, C., Meier, S., Langkilde, A. E., Gotfredsen, C. H., Sattler, M., & Bach, A. (2020). Developing Inhibitors of the p47phox-p22phox Protein-Protein Interaction by Fragment-Based Drug Discovery. Journal of Medicinal Chemistry, 63(3), 1156-1177. https://doi.org/10.1021/acs.jmedchem.9b01492