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Current therapy includes pentavalent antimonial and amphotericin B, but adverse effects persist. The development of a new, more selective, and less toxic combination therapy is therefore a rational and promising approach. In this study, we report the effects of combinations of the monoterpenes limonene (Lim) and carvacrol (Car) with drugs targeting ergosterol biosynthesis, namely nystatin (Nys), tioconazole (Tio), and rosuvastatin (Ros), on\n                    <jats:italic>Leishmania major<\/jats:italic>\n                    , their cytotoxicity on macrophages, and we evaluate their synergism and mechanisms of action. The combinations inhibited the growth of\n                    <jats:italic>L. major<\/jats:italic>\n                    promastigotes, with Lim-Car combined with nystatin (3:2) exhibiting the highest activity, showing an IC\n                    <jats:sub>50<\/jats:sub>\n                    of 2.02\u00a0\u00b5g\/mL. Furthermore, this combination demonstrated greater action against amastigotes (IC\n                    <jats:sub>50<\/jats:sub>\n                    of 0.53\u00a0\u00b5g\/mL) and a high selectivity index (33.60). Low cytotoxicity was observed in murine macrophages (CC\n                    <jats:sub>50<\/jats:sub>\n                    17.81\u00a0\u00b5g\/mL), as well as a low hemolytic potential (CH\n                    <jats:sub>50<\/jats:sub>\n                    of\u2009&gt;\u2009100\u00a0\u00b5g\/mL). The Lim-Car and nystatin (3:2) combination presented a synergistic effect, with a fractional inhibitory concentration index of 0.4. Changes in parasite membrane integrity were also observed, which may be linked to the interaction of nystatin with the enzyme 14-alpha demethylase, along with an increase in TNF-\u03b1 expression and a reduction in IL-10 and IL-6 levels. These results suggest that the combination of Lim-Car with Nys (3:2) may exert a summative effect through multiple biochemical pathways and warrants further investigation as a potential combined therapy with antileishmanial activity.\n                  <\/jats:p>","DOI":"10.1007\/s00210-026-05160-9","type":"journal-article","created":{"date-parts":[[2026,3,16]],"date-time":"2026-03-16T09:40:59Z","timestamp":1773654059000},"page":"12599-12612","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Limonene and carvacrol combined with drugs that inhibit ergosterol synthesis are potent against Leishmania major"],"prefix":"10.1007","volume":"399","author":[{"given":"Rita de C\u00e1ssia Viana","family":"de Carvalho","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Karla Germana","family":"dos Reis Bacelar","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eduardo Lima","family":"Pereira","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bianca Soriano","family":"dos Anjos","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Evellyn Caroline Silva","family":"Melo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Francisco das Chagas","family":"de Souza Cunha","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Paulo S\u00e9rgio","family":"de Araujo Sousa","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jefferson Almeida","family":"Rocha","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Leiz Maria Costa","family":"V\u00e9ras","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Klinger Ant\u00f4nio","family":"da Franca Rodrigues","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maria das Gra\u00e7as","family":"de Freire Medeiros","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daniel Dias Rufino","family":"Arcanjo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michel Mualem de Moraes","family":"Alves","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fernando A\u00e9cio de Amorim","family":"Carvalho","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2026,3,16]]},"reference":[{"key":"5160_CR1","doi-asserted-by":"publisher","first-page":"89","DOI":"10.7705\/BIOMEDICA.4891","volume":"40","author":"IF Abou-El-Naga","year":"2020","unstructured":"Abou-El-Naga IF, Mady RF, Mogahed NMFH (2020) In vitro effectivity of three approved drugs and their synergistic interaction against Leishmania infantum. 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