{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,29]],"date-time":"2026-07-29T11:11:28Z","timestamp":1785323488119,"version":"3.55.0"},"reference-count":24,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2025,4,12]],"date-time":"2025-04-12T00:00:00Z","timestamp":1744416000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"the Russian Science Foundation","award":["19-74-30007"],"award-info":[{"award-number":["19-74-30007"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>NADPH oxidase enzymes (NOXs) are a family of enzymes generating superoxide, which form reactive oxygen species. NOX2 activity is a causative agent for the progression of many diseases: neurodegenerative, cardiovascular, immune dysregulations, and even hereditary diseases and cancer. Administering antioxidants helps in inhibiting NOX2 activity; however, the development of selective inhibitors may provide greater improvement in the therapy of diseases. Here, an optimized synthesis of two most promising NOX2 inhibitors based on the 3-(indolin-6-yl)-4-(N-pyrazole-sulfonamide)-1H-pyrrolo [2,3-b]pyridine structure, namely, GSK2795039 and NCATS-SM7270, and an isomeric derivative of the same class, IMBIOC-1, is reported. The new modified procedures simplify the isolation, reduce byproduct formation, and improve the yields in 0.1\u20131 g scale preparations. Molecular modeling of the structures of NOX2 complexes with inhibitors validated their binding at the same site as NADPH, with IMBIOC-1 forming the largest number of intermolecular interactions with the NOX2 active site. Testing the effects of the compounds on amyloid beta-induced oxidative stress and toxicity in HMC3 microglial cells showed that all three inhibitors completely prevented the pathological amyloid-beta effect. At the same time, NCATS-SM7270 and IMBIOC-1 provided a stronger protective effect on microglial cell survival than GSK2795039, which allowed us to assert the potential of those compounds as neuroprotective agents.<\/jats:p>","DOI":"10.3390\/ijms26083647","type":"journal-article","created":{"date-parts":[[2025,4,14]],"date-time":"2025-04-14T06:18:36Z","timestamp":1744611516000},"page":"3647","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Improved Synthesis of Effective 3-(Indolin-6-yl)-4-(N-pyrazole-sulfonamide)-1H-pyrrolo[2,3-b]pyridine-Based Inhibitors of NADPH Oxidase 2"],"prefix":"10.3390","volume":"26","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3819-2418","authenticated-orcid":false,"given":"Konstantin V.","family":"Potapov","sequence":"first","affiliation":[{"name":"Engelhardt Institute of Molecular Biology of the Russian Academy of Sciences, 119991 Moscow, Russia"},{"name":"Zelinsky Institute of Organic Chemistry of the Russian Academy of Sciences, 119991 Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9331-7319","authenticated-orcid":false,"given":"Dmitry N.","family":"Platonov","sequence":"additional","affiliation":[{"name":"Zelinsky Institute of Organic Chemistry of the Russian Academy of Sciences, 119991 Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4948-6238","authenticated-orcid":false,"given":"Alexander Yu.","family":"Belyy","sequence":"additional","affiliation":[{"name":"Zelinsky Institute of Organic Chemistry of the Russian Academy of Sciences, 119991 Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maxim A.","family":"Novikov","sequence":"additional","affiliation":[{"name":"Engelhardt Institute of Molecular Biology of the Russian Academy of Sciences, 119991 Moscow, Russia"},{"name":"Zelinsky Institute of Organic Chemistry of the Russian Academy of Sciences, 119991 Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3433-7571","authenticated-orcid":false,"given":"Yury V.","family":"Tomilov","sequence":"additional","affiliation":[{"name":"Zelinsky Institute of Organic Chemistry of the Russian Academy of Sciences, 119991 Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1763-300X","authenticated-orcid":false,"given":"Anastasia A.","family":"Anashkina","sequence":"additional","affiliation":[{"name":"Engelhardt Institute of Molecular Biology of the Russian Academy of Sciences, 119991 Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kristina A.","family":"Mukhina","sequence":"additional","affiliation":[{"name":"Engelhardt Institute of Molecular Biology of the Russian Academy of Sciences, 119991 Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Olga I.","family":"Kechko","sequence":"additional","affiliation":[{"name":"Engelhardt Institute of Molecular Biology of the Russian Academy of Sciences, 119991 Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5546-4975","authenticated-orcid":false,"given":"Pavel N.","family":"Solyev","sequence":"additional","affiliation":[{"name":"Engelhardt Institute of Molecular Biology of the Russian Academy of Sciences, 119991 Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Roman A.","family":"Novikov","sequence":"additional","affiliation":[{"name":"Engelhardt Institute of Molecular Biology of the Russian Academy of Sciences, 119991 Moscow, Russia"},{"name":"Zelinsky Institute of Organic Chemistry of the Russian Academy of Sciences, 119991 Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alexander A.","family":"Makarov","sequence":"additional","affiliation":[{"name":"Engelhardt Institute of Molecular Biology of the Russian Academy of Sciences, 119991 Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1517-1983","authenticated-orcid":false,"given":"Vladimir A.","family":"Mitkevich","sequence":"additional","affiliation":[{"name":"Engelhardt Institute of Molecular Biology of the Russian Academy of Sciences, 119991 Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,4,12]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1750","DOI":"10.1111\/bph.13551","article-title":"Redox-based therapeutics in neurodegenerative disease","volume":"174","author":"McBean","year":"2017","journal-title":"Br. J. Pharmacol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1186\/s13024-017-0150-7","article-title":"NADPH oxidase in brain injury and neurodegenerative disorders","volume":"12","author":"Ma","year":"2017","journal-title":"Mol. Neurodegener."},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Mason, H., Rai, G., Kozyr, A., De Jonge, N., Gliniewicz, E., Berg, L.J., Wald, G., Dorrier, C., Henderson, M.J., and Zakharov, A. (2023). Development of an improved and specific inhibitor of NADPH oxidase 2 to treat traumatic brain injury. Redox Biol., 60.","DOI":"10.1016\/j.redox.2023.102611"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"41","DOI":"10.1186\/1742-2094-7-41","article-title":"Gp91phox (NOX2) in classically activated microglia exacerbates traumatic brain injury","volume":"7","author":"Dohi","year":"2010","journal-title":"J Neuroinflamm."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"2409","DOI":"10.1007\/s00018-012-1015-4","article-title":"Targeting microglia-mediated neurotoxicity: The potential of NOX2 inhibitors","volume":"69","author":"Surace","year":"2012","journal-title":"Cell. Mol. Life Sci."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1347","DOI":"10.1073\/pnas.0711568105","article-title":"Nox2-derived radicals contribute to neurovascular and behavioral dysfunction in mice overexpressing the amyloid precursor protein","volume":"105","author":"Park","year":"2008","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"84","DOI":"10.1186\/s13024-017-0225-5","article-title":"NADPH oxidases in Parkinson\u2019s disease: A systematic review","volume":"12","author":"Belarbi","year":"2017","journal-title":"Mol. Neurodegener."},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Reis, J., Massari, M., Marchese, S., Ceccon, M., Aalbers, F.S., Corana, F., Valente, S., Mai, A., Magnani, F., and Mattevi, A. (2020). A closer look into NADPH oxidase inhibitors: Validation and insight into their mechanism of action. Redox Biol., 32.","DOI":"10.1016\/j.redox.2020.101466"},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Liu, F.-C., Yu, H.-P., Chen, P.-J., Yang, H.-W., Chang, S.-H., Tzeng, C.-C., Cheng, W.-J., Chen, Y.-R., Chen, Y.-L., and Hwang, T.-L. (2019). A novel NOX2 inhibitor attenuates human neutrophil oxidative stress and ameliorates inflammatory arthritis in mice. Redox Biol., 26.","DOI":"10.1016\/j.redox.2019.101273"},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Juric, M., Rawat, V., Amaradhi, R., Zielonka, J., and Ganesh, T. (2023). Novel NADPH Oxidase-2 Inhibitors as Potential Anti-Inflammatory and Neuroprotective Agents. Antioxidants, 12.","DOI":"10.3390\/antiox12091660"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"387","DOI":"10.1016\/j.freeradbiomed.2017.08.006","article-title":"NADPH oxidases as drug targets and biomarkers in neurodegenerative diseases: What is the evidence?","volume":"112","author":"Sorce","year":"2017","journal-title":"Free Radic. Biol. Med."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"358","DOI":"10.1089\/ars.2014.6202","article-title":"Discovery of GSK2795039, a Novel Small Molecule NADPH Oxidase 2 Inhibitor","volume":"23","author":"Hirano","year":"2015","journal-title":"Antioxid. Redox Signal."},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Padilha, E.C., Shah, P., Rai, G., and Xu, X. (2021). NOX2 inhibitor GSK2795039 metabolite identification towards drug optimization. J. Pharm. Biomed. Anal., 201.","DOI":"10.1016\/j.jpba.2021.114102"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1582","DOI":"10.4103\/1673-5374.303036","article-title":"P2 \u00d7 7 receptor activation aggravates NADPH oxidase 2-induced oxidative stress after intracerebral hemorrhage","volume":"16","author":"Deng","year":"2021","journal-title":"Neural Regener. Res."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1097","DOI":"10.1007\/s11064-020-02987-3","article-title":"An Effective NADPH Oxidase 2 Inhibitor Provides Neuroprotection and Improves Functional Outcomes in Animal Model of Traumatic Brain Injury","volume":"45","author":"Wang","year":"2020","journal-title":"Neurochem. Res."},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Malkov, A., Popova, I., Ivanov, A., Jang, S.S., Yoon, S.Y., Osypov, A., Huang, Y., Zilberter, Y., and Zilberter, M.A. (2021). \u03b2 initiates brain hypometabolism, network dysfunction and behavioral abnormalities via NOX2-induced oxidative stress in mice. Commun. Biol., 4.","DOI":"10.1038\/s42003-021-02551-x"},{"key":"ref_17","unstructured":"Chen, D.W., Duncan, S., King, N.P., Lee, K.C., Mak, S.Y., and Rivers, D.A. (2012). Novel compounds. (WO2012170752A1)."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"709","DOI":"10.1055\/s-1987-28053","article-title":"Reactions of Sodium Borohydride in Acidic Media; XVI.1 N-Methylation of Amines with Paraformaldehyde\/Trifluoroacetic Acid","volume":"8","author":"Gribble","year":"1987","journal-title":"Synthesis"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"2042","DOI":"10.1002\/cbic.200900199","article-title":"Development of Selective Bisubstrate-Based Inhibitors Against Protein Kinase C (PKC) Isozymes By Using Dynamic Peptide Microarrays","volume":"10","author":"Poot","year":"2009","journal-title":"ChemBioChem"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"443","DOI":"10.1016\/j.bmc.2017.12.001","article-title":"Design. Synthesis and pharmacological evaluation of new acyl sulfonamides as potent and selective Bcl-2 inhibitors","volume":"26","author":"Liu","year":"2018","journal-title":"Bioorg. Med. Chem."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"189","DOI":"10.1038\/s41586-024-07056-1","article-title":"Structure of human phagocyte NADPH oxidase in the activated state","volume":"627","author":"Liu","year":"2024","journal-title":"Nature"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"388","DOI":"10.1016\/S1474-4422(15)70016-5","article-title":"Neuroinflammation in Alzheimer\u2019s disease","volume":"14","author":"Heneka","year":"2015","journal-title":"Lancet Neurol."},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Maurya, S.K., Bhattacharya, N., Mishra, S., Bhattacharya, A., Banerjee, P., Senapati, S., and Mishra, R. (2021). Microglia Specific Drug Targeting Using Natural Products for the Regulation of Redox Imbalance in Neurodegeneration. Front. Pharmacol., 12.","DOI":"10.3389\/fphar.2021.654489"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"3240","DOI":"10.1172\/JCI90606","article-title":"Microglia in Alzheimer\u2019s disease","volume":"127","author":"Sarlus","year":"2017","journal-title":"J. Clin. Investig."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/8\/3647\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T17:13:15Z","timestamp":1760029995000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/8\/3647"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,4,12]]},"references-count":24,"journal-issue":{"issue":"8","published-online":{"date-parts":[[2025,4]]}},"alternative-id":["ijms26083647"],"URL":"https:\/\/doi.org\/10.3390\/ijms26083647","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,4,12]]}}}