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One of the principal mechanisms contributing to AMR is the activity of microbial efflux pumps (EPs), which expel diverse antibiotics out of bacterial cells, thereby rendering them ineffective. Our research aimed to expand the range of molecular classes that inhibit the <jats:italic>Staphylococcus aureus<\/jats:italic> EP NorA. In this study, starting from the hit compound pyrazolo[4,3\u2010<jats:italic>c<\/jats:italic>][1,2]benzothiazine 5,5\u2010dioxide <jats:bold>1<\/jats:bold>, previously reported as a NorA efflux pump inhibitor (EPI), we undertook medicinal chemistry efforts, which involved the iterative combination of the design and synthesis of new analogues with data obtained through ethidium bromide efflux inhibition assays. Subsequent synergistic assays with ciprofloxacin (CPX) against the resistant strain SA\u20101199B led to the identification of three potent compounds (<jats:bold>3<\/jats:bold>, <jats:bold>10<\/jats:bold>, and <jats:bold>19<\/jats:bold>). The evaluation of these compounds in combination with CPX against NorA\u2010overexpressing and NorA\u2010knockout strains (SA\u2010K2378 and SA\u2010K1902, respectively) confirmed that the observed synergy with CPX is dependent on the presence of NorA. Additionally, the combination of NorA EPIs with CPX reduced biofilm production in NorA\u2010overexpressing strains. These findings enhance our understanding of the structure\u2013activity relationship of pyrazolobenzothiazine derivatives and support the use of EtBr efflux assays for rapid NorA inhibitors' identification.<\/jats:p>","DOI":"10.1002\/ardp.70000","type":"journal-article","created":{"date-parts":[[2025,5,20]],"date-time":"2025-05-20T09:46:13Z","timestamp":1747734373000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["NorA Efflux Pump Inhibitors: Expanding SAR Knowledge of Pyrazolo[4,3\u2010<i>c<\/i>][1,2]benzothiazine 5,5\u2010Dioxide Derivatives"],"prefix":"10.1002","volume":"358","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-3591-0210","authenticated-orcid":false,"given":"Giada","family":"Cernicchi","sequence":"first","affiliation":[{"name":"Department of Pharmaceutical Sciences Universit\u00e0 degli Studi di Perugia  Perugia 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