Journal article
Developing Inhibitors of the p47phox-p22phox Protein-Protein Interaction by Fragment-Based Drug Discovery
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.
Language: | English |
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Publisher: | American Chemical Society |
Year: | 2020 |
Pages: | 1156-1177 |
ISSN: | 15204804 and 00222623 |
Types: | Journal article |
DOI: | 10.1021/acs.jmedchem.9b01492 |
ORCIDs: | Meier, Sebastian , 0000-0003-2467-4205 , 0000-0002-1594-0527 , 0000-0003-4305-9910 , 0000-0003-0233-7160 , 0000-0003-2377-1097 , 0000-0001-9164-9961 , 0000-0002-6621-6475 , 0000-0001-7612-223X and Gotfredsen, Charlotte Held |