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Among the species related to human infection, P. falciparum and P. vivax are known for their high incidence and pathogenicity. Despite several approved drugs in the treatment, the increase in resistance mechanisms is becoming increasingly prevalent, which makes the discovery of effective and safer drugs challenging. Thus, it is necessary to explore new mechanisms of action for the discovery of innovative antimalarial agents. Among the explored targets, proteases, especially subtilisin, have shown great promise in the development of new therapeutic options. Method: A narrative review was conducted using the main databases to provide critical information about the subtilisin to design antimalarial drugs. Results: Critical data were found about the isoforms of subtilisins, highlighting SUB1 and SUB2. SBDD approaches were able to show that compounds designed to target the catalytic Asp372, His428, and Ser606, and other such Leu469, Gly467, and Asn520 against SUB1, presented critical results. In addition, quinoline, benzopyran, and triterpene derivatives and peptide inhibitors show their importance, and these scaffolds can be explored in further work. Conclusions: Considering the relevance of this target, this review provided insights into medicinal chemistry, the discovery of antimalarial drugs that act by inhibiting subtilisin, and promoted a promising initiative to combat malaria.<\/jats:p>","DOI":"10.3390\/ph18091318","type":"journal-article","created":{"date-parts":[[2025,9,3]],"date-time":"2025-09-03T08:04:15Z","timestamp":1756886655000},"page":"1318","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Exploring Subtilisin Inhibition to Discover Antimalarial Drugs: Insights into Medicinal Chemistry and Drug Discovery"],"prefix":"10.3390","volume":"18","author":[{"ORCID":"https:\/\/orcid.org\/0009-0004-0694-3306","authenticated-orcid":false,"given":"Margarida","family":"Cochicho Leonardo","sequence":"first","affiliation":[{"name":"Global Health and Tropical Medicine (GHTM), Associate Laboratory in Translation and Innovation Towards Global Health (LA-REAL), Instituto de Higiene e Medicina Tropical (IHMT), Universidade NOVA de Lisboa (UNL), Rua da Junqueira 100, 1349-008 Lisbon, Portugal"}]},{"given":"Sonaly Lima","family":"Albino","sequence":"additional","affiliation":[{"name":"Postgraduate Program of Pharmaceutical Sciences, Department of Pharmacy, State University of Para\u00edba, Campina Grande 58429-500, PB, Brazil"}]},{"given":"Wallyson Junio Santos","family":"de Ara\u00fajo","sequence":"additional","affiliation":[{"name":"Drug Development and Synthesis Laboratory, Department of Pharmacy, State University of Para\u00edba, Campina Grande 58429-500, PB, Brazil"}]},{"given":"Maria Ver\u00f4nica de Barros","family":"Nascimento","sequence":"additional","affiliation":[{"name":"Drug Development and Synthesis Laboratory, Department of Pharmacy, State University of Para\u00edba, Campina Grande 58429-500, PB, Brazil"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5247-867X","authenticated-orcid":false,"given":"Juan David","family":"Rodr\u00edguez-Mac\u00edas","sequence":"additional","affiliation":[{"name":"Facultad de Ciencias de la Salud, Exactas y Naturales, Universidad Libre Seccional Barranquilla, Barranquilla 080001, Colombia"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7503-1528","authenticated-orcid":false,"given":"Edgar Alexander Marquez","family":"Brazon","sequence":"additional","affiliation":[{"name":"Facultad de Ciencias B\u00e1sicas, Departamento de Qu\u00edmica y Biolog\u00eda, Universidad del Norte, Barranquilla 081007, Colombia"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1590-3750","authenticated-orcid":false,"given":"Ricardo Olimpio de","family":"Moura","sequence":"additional","affiliation":[{"name":"Postgraduate Program of Pharmaceutical Sciences, Department of Pharmacy, State University of Para\u00edba, Campina Grande 58429-500, PB, Brazil"},{"name":"Drug Development and Synthesis Laboratory, Department of Pharmacy, State University of Para\u00edba, Campina Grande 58429-500, PB, Brazil"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0313-0778","authenticated-orcid":false,"given":"F\u00e1tima","family":"Nogueira","sequence":"additional","affiliation":[{"name":"Global Health and Tropical Medicine (GHTM), Associate Laboratory in Translation and Innovation Towards Global Health (LA-REAL), Instituto de Higiene e Medicina Tropical (IHMT), Universidade NOVA de Lisboa (UNL), Rua da Junqueira 100, 1349-008 Lisbon, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2664-4336","authenticated-orcid":false,"given":"Igor Jos\u00e9 dos Santos","family":"Nascimento","sequence":"additional","affiliation":[{"name":"Global Health and Tropical Medicine (GHTM), Associate Laboratory in Translation and Innovation Towards Global Health (LA-REAL), Instituto de Higiene e Medicina Tropical (IHMT), Universidade NOVA de Lisboa (UNL), Rua da Junqueira 100, 1349-008 Lisbon, Portugal"},{"name":"Postgraduate Program of Pharmaceutical Sciences, Department of Pharmacy, State University of Para\u00edba, Campina Grande 58429-500, PB, Brazil"},{"name":"Drug Development and Synthesis Laboratory, Department of Pharmacy, State University of Para\u00edba, Campina Grande 58429-500, PB, Brazil"}]}],"member":"1968","published-online":{"date-parts":[[2025,9,3]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"256","DOI":"10.1016\/j.it.2023.02.005","article-title":"Malaria: Influence of Anopheles Mosquito Saliva on Plasmodium Infection","volume":"44","author":"Arora","year":"2023","journal-title":"Trends Immunol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1601","DOI":"10.1080\/17460913.2024.2412397","article-title":"Current Trends to Design Antimalarial Drugs Targeting N-Myristoyltransferase","volume":"19","author":"Weber","year":"2024","journal-title":"Future Microbiol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"115550","DOI":"10.1016\/j.ejmech.2023.115550","article-title":"Exploring N-Myristoyltransferase as a Promising 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