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and severity of fungal infections and the resistance to available antifungals highlights the imperative need for novel therapeutics and the search for new targets. High-content screening of libraries containing hundreds of compounds is a powerful strategy for searching for new drug candidates. In this study, we screened the Pandemic Response Box library (Medicines for Malaria Venture) of 400 diverse molecules against the Sporothrix pathogenic species. The initial screen identified twenty-four candidates that inhibited the growth of Sporothrix brasiliensis by more than 80%. Some of these compounds are known to display antifungal activity, including olorofim (MMV1782354), a new antifungal drug. Olorofim inhibited and killed the yeasts of S. brasiliensis, S. schenckii, and S. globosa at concentrations lower than itraconazole, and it also showed antibiofilm activity. According to the results obtained by fluorimetry, electron microscopy, and particle characterization analyses, we observed that olorofim induced profound alterations on the cell surface and cell cycle arrest in S. brasiliensis yeasts. We also verified that these morphophysiological alterations impaired their ability to adhere to keratinocytes in vitro. Our results indicate that olorofim is a promising new antifungal against sporotrichosis agents.<\/jats:p>","DOI":"10.3390\/jof8101004","type":"journal-article","created":{"date-parts":[[2022,9,26]],"date-time":"2022-09-26T01:41:58Z","timestamp":1664156518000},"page":"1004","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":17,"title":["Screening of Pandemic Response Box Library Reveals the High Activity of Olorofim against Pathogenic Sporothrix Species"],"prefix":"10.3390","volume":"8","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-6576-6646","authenticated-orcid":false,"given":"Luana Pereira","family":"Borba-Santos","sequence":"first","affiliation":[{"name":"Laborat\u00f3rio de Biologia Celular de Fungos, Instituto de Biof\u00edsica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Rio de Janeiro 21941-902, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8316-4670","authenticated-orcid":false,"given":"Rodrigo","family":"Rollin-Pinheiro","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio de Qu\u00edmica Biol\u00f3gica de Microrganismos, Departamento de Microbiologia Geral, Instituto de Microbiologia Paulo de G\u00f3es, Universidade Federal do Rio de Janeiro, Rio de Janeiro 21941-902, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8938-9267","authenticated-orcid":false,"given":"Yasmin","family":"da Silva Fontes","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio de Bioqu\u00edmica Microbiana, Departamento de Microbiologia Geral, Instituto de Microbiologia Paulo de G\u00f3es, Universidade Federal do Rio de Janeiro, Rio de Janeiro 21941-902, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Giulia Maria Pires","family":"dos Santos","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio de Biologia Celular de Fungos, Instituto de Biof\u00edsica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Rio de Janeiro 21941-902, Brazil"},{"name":"Laborat\u00f3rio de Qu\u00edmica Biol\u00f3gica de Microrganismos, Departamento de Microbiologia Geral, Instituto de Microbiologia Paulo de G\u00f3es, Universidade Federal do Rio de Janeiro, Rio de Janeiro 21941-902, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5478-9006","authenticated-orcid":false,"given":"Glauber Ribeiro","family":"de Sousa Ara\u00fajo","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio de Biof\u00edsica de Fungos, Instituto de Biof\u00edsica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Rio de Janeiro 21941-902, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0891-4563","authenticated-orcid":false,"given":"Anderson Messias","family":"Rodrigues","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio de Pat\u00f3genos F\u00fangicos Emergentes, Departamento de Microbiologia, Imunologia e Parasitologia, Universidade Federal de S\u00e3o Paulo, S\u00e3o Paulo 04023-062, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2856-9667","authenticated-orcid":false,"given":"Allan J.","family":"Guimar\u00e3es","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio de Bioqu\u00edmica e Imunologia das Micoses, Departamento de Microbiologia e Parasitologia, Instituto Biom\u00e9dico, Universidade Federal Fluminense, Niter\u00f3i 24210-130, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Wanderley","family":"de Souza","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio de Ultraestrutura Celular Hertha Meyer, Instituto de Biof\u00edsica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Rio de Janeiro 21941-902, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Susana","family":"Frases","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio de Biof\u00edsica de Fungos, Instituto de Biof\u00edsica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Rio de Janeiro 21941-902, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0474-8388","authenticated-orcid":false,"given":"Antonio","family":"Ferreira-Pereira","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio de Bioqu\u00edmica Microbiana, Departamento de Microbiologia Geral, Instituto de Microbiologia Paulo de G\u00f3es, Universidade Federal do Rio de Janeiro, Rio de Janeiro 21941-902, 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