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These species are considered resistant fungi due to their low susceptibility to most current antifungal agents used in healthcare settings. The search for new compounds that could work as promising candidate antifungal drugs is an increasing field of interest. In this context, in the present study we screened the Pandemic Response Box<jats:sup>\u00ae<\/jats:sup> library (Medicines for Malaria Venture [MMV], Switzerland) to identify compounds with antifungal activity against <jats:italic>Scedosporium<\/jats:italic> and <jats:italic>Lomentospora<\/jats:italic> species. An initial screening of the drugs from this collection at 5 \u03bcM was performed using a clinical <jats:italic>Scedosporium aurantiacum<\/jats:italic> isolate according to the EUCAST protocol. Compounds with activity against this fungus were also tested against four other species (<jats:italic>S<\/jats:italic>. <jats:italic>boydii<\/jats:italic>\u00b8 <jats:italic>S<\/jats:italic>. <jats:italic>dehoogii<\/jats:italic>, <jats:italic>S<\/jats:italic>. <jats:italic>apiospermum<\/jats:italic> and <jats:italic>L<\/jats:italic>. <jats:italic>prolificans<\/jats:italic>) at concentrations ranging from 0.078 to 10 \u03bcM. Seven compounds inhibited more than 80% of <jats:italic>S<\/jats:italic>. <jats:italic>aurantiacum<\/jats:italic> growth, three of them (alexidine, amorolfine and olorofim) were selected due to their differences in mechanism of action, especially when compared to drugs from the azole class. These compounds were more active against biofilm formation than against preformed biofilm in <jats:italic>Scedosporium<\/jats:italic> and <jats:italic>Lomentospora<\/jats:italic> species, except alexidine, which was able to decrease preformed biofilm about 50%. Analysis of the potential synergism of these compounds with voriconazole and caspofungin was performed by the checkerboard method for <jats:italic>S<\/jats:italic>. <jats:italic>aurantiacum<\/jats:italic>. The analysis by Bliss methodology revealed synergistic effects among selected drugs with caspofungin. When these drugs were combined with voriconazole, only alexidine and amorolfine showed a synergistic effect, whereas olorofim showed an antagonistic effect. Scanning electron microscopy revealed that alexidine induces morphology alterations in <jats:italic>S<\/jats:italic>. <jats:italic>aurantiacum<\/jats:italic> biofilm grown on a catheter surface. Reactive oxygen species production, mitochondrial activity and surface components were analyzed by fluorescent probes when <jats:italic>S<\/jats:italic>. <jats:italic>aurantiacum<\/jats:italic> was treated with selected drugs and revealed that some cell parameters are altered by these compounds. In conclusion, alexidine, amorolfine and olorofim were identified as promising compounds to be studied against scedosporiosis and lomentosporiosis.<\/jats:p>","DOI":"10.1371\/journal.pone.0280964","type":"journal-article","created":{"date-parts":[[2023,2,3]],"date-time":"2023-02-03T18:35:27Z","timestamp":1675449327000},"page":"e0280964","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":9,"title":["Pandemic Response Box\u00ae library as a source of antifungal drugs against Scedosporium and Lomentospora species"],"prefix":"10.1371","volume":"18","author":[{"given":"Rodrigo","family":"Rollin-Pinheiro","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1803-6381","authenticated-orcid":true,"given":"Mariana Ingrid Dutra da Silva","family":"Xisto","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuri","family":"de Castro-Almeida","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Victor Pereira","family":"Rochetti","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6576-6646","authenticated-orcid":true,"given":"Luana Pereira","family":"Borba-Santos","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8938-9267","authenticated-orcid":true,"given":"Yasmin da Silva","family":"Fontes","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Antonio","family":"Ferreira-Pereira","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sonia","family":"Rozental","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8944-9415","authenticated-orcid":true,"given":"Eliana","family":"Barreto-Bergter","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2023,2,3]]},"reference":[{"issue":"1","key":"pone.0280964.ref001","doi-asserted-by":"crossref","first-page":"157","DOI":"10.1128\/CMR.00039-07","article-title":"Infections caused by Scedosporium spp","volume":"21","author":"KJ Cortez","year":"2008","journal-title":"Clin Microbiol Rev"},{"issue":"4","key":"pone.0280964.ref002","doi-asserted-by":"crossref","first-page":"371","DOI":"10.1080\/13693780802256539","article-title":"Different virulence of the species of the Pseudallescheria boydii complex","volume":"47","author":"F Gilgado","year":"2009","journal-title":"Med Mycol"},{"issue":"2","key":"pone.0280964.ref003","doi-asserted-by":"crossref","first-page":"249","DOI":"10.1016\/j.mycmed.2018.02.015","article-title":"Pseudallescheria\/Scedosporium complex species: From saprobic to pathogenic fungus","volume":"28","author":"N. 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