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Despite its great medical importance, there are few treatments available and these are associated with adverse events and resistance. In this work, after screening the drugs present in the Medicines for Malaria Venture Pandemic Box, we found new hits with anti-Toxoplasma gondii activity. Through our analysis, we selected twenty-three drugs or drug-like compounds that inhibited the proliferation of T. gondii tachyzoites in vitro by more than 50% at a concentration of 1 \u00b5M after seven days of treatment. Nineteen of these compounds have never been reported active before against T. gondii. Inhibitory curves showed that most of these drugs were able to inhibit parasite replication with IC50 values on the nanomolar scale. To better understand the unprecedented effect of seven compounds against T. gondii tachyzoites, an ultrastructural analysis was carried out using transmission electron microscopy. Treatment with 0.25 \u00b5M verdinexor, 3 nM MMV1580844, and 0.25 \u00b5M MMV019724 induced extensive vacuolization, complete ultrastructural disorganization, and lytic effects in the parasite, respectively, and all of them showed alterations in the division process. Treatment with 1 \u00b5M Eberconazole, 0.5 \u00b5M MMV1593541, 1 \u00b5M MMV642550, 1 \u00b5M RWJ-67657, and 1 \u00b5M URMC-099-C also caused extensive vacuolization in the parasite. The activity of these drugs against intracellular tachyzoites supports the idea that the drugs selected in the Pandemic Box could be potential future drugs for the treatment of acute toxoplasmosis.<\/jats:p>","DOI":"10.3390\/tropicalmed8120510","type":"journal-article","created":{"date-parts":[[2023,11,29]],"date-time":"2023-11-29T12:01:00Z","timestamp":1701259260000},"page":"510","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Medicines for Malaria Venture Pandemic Box In Vitro Screening Identifies Compounds Highly Active against the Tachyzoite Stage of Toxoplasma gondii"],"prefix":"10.3390","volume":"8","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-3884-0131","authenticated-orcid":false,"given":"Mike","family":"dos Santos","sequence":"first","affiliation":[{"name":"Laborat\u00f3rio de Quimioterapia de Protozo\u00e1rios Egler Chiari, Departamento de Parasitologia, Instituto de Ci\u00eancias Biol\u00f3gicas, Universidade Federal de Minas Gerais, Belo Horizonte 31270-901, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andr\u00e9ia Luiza","family":"Oliveira Costa","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio de Quimioterapia de Protozo\u00e1rios Egler Chiari, Departamento de Parasitologia, Instituto de Ci\u00eancias Biol\u00f3gicas, Universidade Federal de Minas Gerais, Belo Horizonte 31270-901, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Guilherme Henrique de Souza","family":"Vaz","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio de Quimioterapia de Protozo\u00e1rios Egler Chiari, Departamento de Parasitologia, Instituto de Ci\u00eancias Biol\u00f3gicas, Universidade Federal de Minas Gerais, Belo Horizonte 31270-901, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gabriela Carolina Alves","family":"de Souza","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio de Quimioterapia de Protozo\u00e1rios Egler Chiari, Departamento de Parasitologia, Instituto de Ci\u00eancias Biol\u00f3gicas, Universidade Federal de Minas Gerais, Belo Horizonte 31270-901, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8599-0474","authenticated-orcid":false,"given":"Ricardo Wagner de Almeida","family":"Vitor","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio de Toxoplasmose, Departamento de Parasitologia, Instituto de Ci\u00eancias Biol\u00f3gicas, Universidade Federal de Minas Gerais, Belo Horizonte 31270-901, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0925-6953","authenticated-orcid":false,"given":"\u00c9rica S.","family":"Martins-Duarte","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio de Quimioterapia de Protozo\u00e1rios Egler Chiari, Departamento de Parasitologia, Instituto de Ci\u00eancias Biol\u00f3gicas, Universidade Federal de Minas Gerais, Belo Horizonte 31270-901, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,11,29]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Crabtree-Ram\u00edrez, B., Caro-Vega, Y., Shepherd, B.E., Grinsztejn, B., Wolff, M., Cortes, C.P., Padgett, D., Carriquiry, G., Fink, V., and Jayathilake, K. 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