A Novel and Specific NADPH Oxidase-1 (Nox1) Small-Molecule Inhibitor Blocks the Formation of Functional Invadopodia in Human Colon Cancer Cells
Open accessAbstract
The NADPH oxidase (Nox) proteins catalyze the regulated formation of reactive oxygen species (ROS), which play key roles as signaling molecules in several physiological and pathophysiological processes. ROS generation by the Nox1 member of the Nox family is necessary for the formation of extracellular matrix (ECM)-degrading, actin-rich cellular structures known as invadopodia. Selective inhibition of Nox isoforms can provide reversible, mechanistic insights into these cellular processes in contrast to scavenging or inhibition of ROS production. Currently no specific Nox inhibitors have been described. Here, by high-throughput screening, we identify a subset of phenothiazines, 2-acetylphenothiazine (here referred to as ML171) (and its related 2-(trifluoromethyl)-phenothiazine) as nanomolar, cell-active, and specific Nox1 inhibitors that potently block Nox1-dependent ROS generation, with only marginal activity on other cellular ROS-producing enzymes and receptors including the other Nox isoforms. ML171 also blocks the ROS-dependent formation of ECM-degrading invadopodia in colon cancer cells. Such effects can be reversed by overexpression of Nox1 protein, which is suggestive of a selective mechanism of inhibition of Nox1 by this compound. These results elucidate the relevance of Nox1-dependent ROS generation in mechanisms of cancer invasion and define ML171 as a useful Nox1 chemical probe and potential therapeutic agent for inhibition of cancer cell invasion.
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ML171 inhibits NOX1 at nanomolar concentrations, with IC50 values at least 20-fold higher for NOX2, NOX3, NOX4, and XO.
“ML171 inhibits NOX1 at nanomolar concentrations, with IC50 values at least 20-fold higher for NOX2, NOX3, NOX4, and XO (359).”
From Sources of Vascular Nitric Oxide and Reactive Oxygen Species and Their Regulation (2019) · cites this paper as [359]
ML171 showed off-target effects with the inhibition of serotonin and adrenergic receptors in the micromolar range.
“ML171 showed off-target effects with the inhibition of serotonin and adrenergic receptors in the micromolar range (359).”
From Sources of Vascular Nitric Oxide and Reactive Oxygen Species and Their Regulation (2019) · cites this paper as [359]
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- Authors
- Gianni, D., Taulet, N., Zhang, H., DerMardirossian, C., Kister, J., Martinez, L., Roush, W. R., Brown, S. J., Bokoch, G. M., & Rosen, H.
- Affiliations
- Scripps Research Institute (US)
- University of California, San Francisco (US)
- Department of Chemistry, The Scripps Research Institute, Jupiter, Florida 33458
- Corresponding author
- Rosen, Hugh
- Venue
- ACS Chemical Biology
- Publisher
- American Chemical Society (ACS)
- Published
- 2010-08-26
- Volume
- 5
- Issue
- 10
- Pages
- 981-993
- ISSN
- 1554-8929 (ISSN-L)
- 1554-8937
- PMC
- PMC2955773
- OpenAlex
- W1976254783
- Cited by
- 192
- References
- 43
- Citations by year
- 2026: 3 · 2025: 8 · 2024: 8 · 2023: 7 · 2022: 5 · 2021: 12 · 2020: 20 · 2019: 15 · 2018: 12 · 2017: 10 · 2016: 15 · 2015: 18 · 2014: 18 · 2013: 16 · 2012: 18
- FWCI
- 5.35
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- ACS Chemical Biology (publishedVersion)
- PubMed (publishedVersion)
- Europe PMC (PubMed Central) (acceptedVersion)
- PubMed Central (acceptedVersion)
- Funders
- National Institutes of Health (5u54mh074404-03)
- U.S. Public Health Service (HHSN-271-2008-00025-C)
- National Heart, Lung, and Blood Institute (HL-48008, R01HL048008)
- National Institute of Mental Health (U54MH084512, 5U54MH074404, U54-MH074404)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Redox biology and oxidative stress
- Life Sciences › Immunology and Microbiology › Immunology › Neutrophil, Myeloperoxidase and Oxidative Mechanisms
- Life Sciences › Pharmacology, Toxicology and Pharmaceutics › Toxicology › Bioactive Compounds and Antitumor Agents
- Keywords
- NOX1, invadopodia, reactive oxygen species, colon cancer, tumor invasion, phenothiazines, extracellular matrix degradation, high-throughput screening, selective enzyme inhibition
- MeSH terms
- NADPH Oxidase 1, Antineoplastic Agents, Colonic Neoplasms, Humans, NADH, NADPH Oxidoreductases, Neoplasm Invasiveness, Phenothiazines, Reactive Oxygen Species, Cell Line, Tumor, Small Molecule Libraries, High-Throughput Screening Assays
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