{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,11]],"date-time":"2026-03-11T09:35:56Z","timestamp":1773221756785,"version":"3.50.1"},"reference-count":26,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2025,1,23]],"date-time":"2025-01-23T00:00:00Z","timestamp":1737590400000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2025,1,23]],"date-time":"2025-01-23T00:00:00Z","timestamp":1737590400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"DOI":"10.13039\/501100004586","name":"Funda\u00e7\u00e3o Carlos Chagas Filho de Amparo \u00e0 Pesquisa do Estado do Rio de Janeiro","doi-asserted-by":"publisher","award":["E-26\/202 421\/2017"],"award-info":[{"award-number":["E-26\/202 421\/2017"]}],"id":[{"id":"10.13039\/501100004586","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004586","name":"Funda\u00e7\u00e3o Carlos Chagas Filho de Amparo \u00e0 Pesquisa do Estado do Rio de Janeiro","doi-asserted-by":"publisher","award":["E-26\/202.942\/2019"],"award-info":[{"award-number":["E-26\/202.942\/2019"]}],"id":[{"id":"10.13039\/501100004586","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003593","name":"Conselho Nacional de Desenvolvimento Cient\u00edfico e Tecnol\u00f3gico","doi-asserted-by":"publisher","award":["PQ-2 (304712\/2016-7)"],"award-info":[{"award-number":["PQ-2 (304712\/2016-7)"]}],"id":[{"id":"10.13039\/501100003593","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100002322","name":"Coordena\u00e7\u00e3o de Aperfei\u00e7oamento de Pessoal de N\u00edvel Superior","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100002322","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Parasites Vectors"],"abstract":"<jats:title>Abstract<\/jats:title>\n          <jats:sec>\n            <jats:title>Background<\/jats:title>\n            <jats:p>Leishmaniases are neglected tropical diseases with great clinical and epidemiological importance. The current chemotherapy available for the treatment of leishmaniasis\u00a0presents  several problems, such as adverse effects, toxicity, long treatment time, and parasite resistance. The discovery of new therapeutic alternatives is extremely essential, and the discovery of cellular targets is a tool that helps in the development of new drugs. Serine proteases emerge as important virulence factors in the <jats:italic>Leishmania<\/jats:italic> genus, as they participate in important processes involved in their infectivity, virulence, and survival. In this work, we evaluated the leishmanicidal effect of different serine protease inhibitors (Benzamidine, PF-429242, PMSF, TLCK, and TPCK). Additionally, we determined the implication of pretreatment with these inhibitors on the entry and survival of parasites within macrophages, as well as the conversion of promastigotes into amastigotes, to discover the importance of serine proteases in the establishment of infection and, consequently, as targets for new drugs for <jats:italic>Leishmania<\/jats:italic>. <\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Results<\/jats:title>\n            <jats:p>In general, the inhibitors had low toxicity in host macrophages, and three showed some effect in promastigote and amastigote forms of <jats:italic>L. amazonensis<\/jats:italic> (PF-429242, TLCK, and TPCK). Using a short incubation interval, we pretreated <jats:italic>L. amazonensis<\/jats:italic> promastigotes with these five compounds before in vitro infection. Pretreatment with PF-429242, TLCK, and TPCK considerably compromised the survival of these parasites inside host macrophages, without altering the entry of promastigotes into these cells and differentiation into amastigotes. In addition, treatment with PF-429242 and TPCK was able to reduce the serine proteases\u2019 enzymatic activity using subtilisin substrate on <jats:italic>L. amazonensis<\/jats:italic> promastigote lysate. <\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Conclusions<\/jats:title>\n            <jats:p>This work highlights the importance of serine proteases in <jats:italic>L. amazonensis<\/jats:italic> as a possible target for new therapeutic alternatives in <jats:italic>Leishmania<\/jats:italic> spp.<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Graphical Abstract<\/jats:title>\n          <\/jats:sec>","DOI":"10.1186\/s13071-024-06630-w","type":"journal-article","created":{"date-parts":[[2025,1,23]],"date-time":"2025-01-23T18:57:26Z","timestamp":1737658646000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Pretreatment with serine protease inhibitors impairs Leishmania amazonensis survival on macrophages"],"prefix":"10.1186","volume":"18","author":[{"given":"Patr\u00edcia","family":"de Almeida Machado","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Pollyanna Stephanie","family":"Gomes","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Elaine Soares","family":"Coimbra","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Herbert Leonel","family":"de Matos Guedes","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2025,1,23]]},"reference":[{"key":"6630_CR1","doi-asserted-by":"publisher","first-page":"1000972","DOI":"10.3389\/fcimb.2022.1000972","volume":"12","author":"DB Scariot","year":"2022","unstructured":"Scariot DB, Staneviciute A, Zhu J, Li X, Scott EA, Engman DM. Leishmaniasis and Chagas disease: is there hope in nanotechnology to fight neglected tropical diseases? Front Cell Infect Microbiol. 2022;12:1000972. https:\/\/doi.org\/10.3389\/fcimb.2022.1000972.","journal-title":"Front Cell Infect Microbiol"},{"key":"6630_CR2","unstructured":"WHO. 2023. https:\/\/www.who.int\/healthtopics\/leishmaniasis#tab=tab_1. Accessed 29 Apr 2024."},{"key":"6630_CR3","doi-asserted-by":"publisher","first-page":"823","DOI":"10.1007\/s40257-022-00726-8","volume":"23","author":"HJC de Vries","year":"2022","unstructured":"de Vries HJC, Schallig HD. Cutaneous leishmaniasis: a 2022 updated narrative review into diagnosis and management developments. Am J Clin Dermatol. 2022;23:823\u201340. https:\/\/doi.org\/10.1007\/s40257-022-00726-8.","journal-title":"Am J Clin Dermatol"},{"key":"6630_CR4","doi-asserted-by":"publisher","first-page":"141","DOI":"10.1177\/2156587215623126","volume":"22","author":"M Foroutan-Rad","year":"2017","unstructured":"Foroutan-Rad M, Tappeh KH, Khademvatan S. Antileishmanial and immunomodulatory activity of Allium sativum (Garlic): a review. J Evid Based Complementary Altern Med. 2017;22:141\u201355. https:\/\/doi.org\/10.1177\/2156587215623126.","journal-title":"J Evid Based Complementary Altern Med"},{"key":"6630_CR5","doi-asserted-by":"publisher","first-page":"2604","DOI":"10.1111\/febs.14130","volume":"284","author":"E Deu","year":"2017","unstructured":"Deu E. Proteases as antimalarial targets: strategies for genetic, chemical, and therapeutic validation. FEBS J. 2017;284:2604\u201328. https:\/\/doi.org\/10.1111\/febs.14130.","journal-title":"FEBS J"},{"key":"6630_CR6","doi-asserted-by":"publisher","first-page":"163","DOI":"10.1016\/j.lfs.2019.01.015","volume":"219","author":"PA Machado","year":"2019","unstructured":"Machado PA, Carneiro MPD, Sousa-Batista AJ, Lopes FJP, Lima A, Chaves SP, et\u00a0al. Leishmanicidal therapy targeted to parasite proteases. Life Sci. 2019;219:163\u201381. https:\/\/doi.org\/10.1016\/j.lfs.2019.01.015.","journal-title":"Life Sci"},{"key":"6630_CR7","doi-asserted-by":"publisher","first-page":"95","DOI":"10.2147\/HIV.S79956","volume":"7","author":"Z Lv","year":"2015","unstructured":"Lv Z, Chu Y, Wang Y. HIV protease inhibitors: a review of molecular selectivity and toxicity. HIV AIDS. 2015;7:95\u2013104. https:\/\/doi.org\/10.2147\/HIV.S79956.","journal-title":"HIV AIDS"},{"key":"6630_CR8","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1016\/0076-6879(94)44003-4","volume":"244","author":"AJ Barrett","year":"1994","unstructured":"Barrett AJ. Classification of peptidases. Methods Enzymol. 1994;244:1\u201315. https:\/\/doi.org\/10.1016\/0076-6879(94)44003-4.","journal-title":"Methods Enzymol"},{"key":"6630_CR9","doi-asserted-by":"publisher","first-page":"3103","DOI":"10.2174\/0929867311320250006","volume":"20","author":"IM Bastos","year":"2013","unstructured":"Bastos IM, Motta FN, Grellier P, Santana JM. Parasite prolyl oligopeptidases and the challenge of designing chemotherapeuticals for Chagas disease, leishmaniasis and African trypanosomiasis. Curr Med Chem. 2013;20:3103\u201315. https:\/\/doi.org\/10.2174\/0929867311320250006.","journal-title":"Curr Med Chem"},{"key":"6630_CR10","doi-asserted-by":"publisher","first-page":"31120","DOI":"10.1074\/jbc.M110.114462","volume":"285","author":"RK Swenerton","year":"2010","unstructured":"Swenerton RK, Knudsen GM, Sajid M, Kelly BL, McKerrow JH. Leishmania subtilisin is a maturase for the trypanothione reductase system and contributes to disease pathology. J Biol Chem. 2010;285:31120\u20139. https:\/\/doi.org\/10.1074\/jbc.M110.114462.","journal-title":"J Biol Chem"},{"key":"6630_CR11","doi-asserted-by":"publisher","first-page":"135","DOI":"10.1016\/j.exppara.2010.04.009","volume":"126","author":"T Taheri","year":"2010","unstructured":"Taheri T, Salmanian AH, Gholami E, Doustdari F, Zahedifard F, Rafati S. Leishmania major: disruption of signal peptidase type I and its consequences on survival, growth and infectivity. Exp Parasitol. 2010;126:135\u201345. https:\/\/doi.org\/10.1016\/j.exppara.2010.04.009.","journal-title":"Exp Parasitol"},{"key":"6630_CR12","doi-asserted-by":"publisher","first-page":"131","DOI":"10.1016\/0003-2697(82)90118-x","volume":"126","author":"LC Green","year":"1982","unstructured":"Green LC, Wagner DA, Glogowski J, Skipper PL, Wishnok JS, Tannenbaum SR. Analysis of nitrate, nitrite, and [15N]nitrate in biological fluids. Anal Biochem. 1982;126:131\u20138. https:\/\/doi.org\/10.1016\/0003-2697(82)90118-x.","journal-title":"Anal Biochem"},{"key":"6630_CR13","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1016\/s0166-6851(01)00290-0","volume":"116","author":"LC Alves","year":"2001","unstructured":"Alves LC, Judice WA, St Hilaire PM, Meldal M, Sanderson SJ, Mottram JC, et\u00a0al. Substrate specificity of recombinant cysteine proteinase, CPB, of Leishmania mexicana. Mol Biochem Parasitol. 2001;116:1\u20139. https:\/\/doi.org\/10.1016\/s0166-6851(01)00290-0.","journal-title":"Mol Biochem Parasitol"},{"key":"6630_CR14","doi-asserted-by":"publisher","first-page":"835910","DOI":"10.1155\/2015\/835910","volume":"2015","author":"IA Rodrigues","year":"2015","unstructured":"Rodrigues IA, Mazotto AM, Cardoso V, Alves RL, Amaral AC, Silva JR, et\u00a0al. Natural products: insights into Leishmaniasis inflammatory response. Mediators Inflamm. 2015;2015:835910. https:\/\/doi.org\/10.1155\/2015\/835910.","journal-title":"Mediators Inflamm"},{"key":"6630_CR15","doi-asserted-by":"publisher","first-page":"67","DOI":"10.1016\/j.exppara.2017.11.008","volume":"184","author":"CR Alves","year":"2018","unstructured":"Alves CR, Souza RS, Charret KDS, Cortes LMC, Sa-Silva MP, Barral-Veloso L, et\u00a0al. Understanding serine proteases implications on Leishmania spp lifecycle. Exp Parasitol. 2018;184:67\u201381. https:\/\/doi.org\/10.1016\/j.exppara.2017.11.008.","journal-title":"Exp Parasitol"},{"key":"6630_CR16","doi-asserted-by":"publisher","first-page":"853","DOI":"10.1007\/s00436-007-0552-5","volume":"101","author":"HL de Matos Guedes","year":"2007","unstructured":"de Matos Guedes HL, Carneiro MP, Gomes DC, Rossi-Bergmanmn B. Giovanni de Simone S. Oligopeptidase B from L. amazonensis. Parasitol Res. 2007;101:853\u201363. https:\/\/doi.org\/10.1007\/s00436-007-0552-5.","journal-title":"Parasitol Res."},{"key":"6630_CR17","doi-asserted-by":"publisher","first-page":"197","DOI":"10.2478\/s11686-008-0026-7","volume":"53","author":"H Matos Guedes","year":"2008","unstructured":"Matos Guedes H, Carvalho R, Oliveira Gomes D, Rossi-Bergmann B, De-Simone SG. Oligopeptidase B-2 from Leishmania amazonensis with an unusual C-terminal extension. Acta Parasitol. 2008;53:197\u2013204. https:\/\/doi.org\/10.2478\/s11686-008-0026-7.","journal-title":"Acta Parasitol"},{"key":"6630_CR18","doi-asserted-by":"publisher","first-page":"2089","DOI":"10.3390\/cimb44050141","volume":"44","author":"PS Gomes","year":"2022","unstructured":"Gomes PS, Carneiro MPD, Machado PA, de Andrade-Neto VV, da Fonseca-Martins AM, Goundry A, et\u00a0al. Subtilisin of Leishmania amazonensis as potential druggable target: subcellular localization, in vitro leishmanicidal activity and molecular docking of PF-429242, a subtilisin inhibitor. Curr Issues Mol Biol. 2022;44:2089\u2013106. https:\/\/doi.org\/10.3390\/cimb44050141.","journal-title":"Curr Issues Mol Biol"},{"key":"6630_CR19","doi-asserted-by":"publisher","DOI":"10.1128\/spectrum.02960-23","volume":"11","author":"EO Porta","year":"2023","unstructured":"Porta EO, Gao L, Denny PW, Steel PG, Kalesh K. Inhibition of HSP90 distinctively modulates the global phosphoproteome of Leishmania mexicana developmental stages. Microbiol Spectr. 2023;11:e0296023. https:\/\/doi.org\/10.1128\/spectrum.02960-23.","journal-title":"Microbiol Spectr"},{"key":"6630_CR20","doi-asserted-by":"publisher","first-page":"13","DOI":"10.1016\/j.molbiopara.2003.12.011","volume":"135","author":"S Rafati","year":"2004","unstructured":"Rafati S, Salmanian AH, Taheri T, Masina S, Schaff C, Taslimi Y, et\u00a0al. Type I signal peptidase from Leishmania is a target of the immune response in human cutaneous and visceral leishmaniasis. Mol Biochem Parasitol. 2004;135:13\u201320. https:\/\/doi.org\/10.1016\/j.molbiopara.2003.12.011.","journal-title":"Mol Biochem Parasitol"},{"key":"6630_CR21","doi-asserted-by":"publisher","first-page":"1910","DOI":"10.1128\/AAC.04742-14","volume":"59","author":"F Souza-Silva","year":"2015","unstructured":"Souza-Silva F, Bourguignon SC, Pereira BA, Cortes LM, de Oliveira LF, Henriques-Pons A, et\u00a0al. Epoxy-alpha-lapachone has in vitro and in vivo anti-Leishmania (Leishmania ) amazonensis effects and inhibits serine proteinase activity in this parasite. Antimicrob Agents Chemother. 2015;59:1910\u20138. https:\/\/doi.org\/10.1128\/AAC.04742-14.","journal-title":"Antimicrob Agents Chemother"},{"key":"6630_CR22","doi-asserted-by":"publisher","first-page":"583834","DOI":"10.3389\/fmicb.2021.583834","volume":"12","author":"PA Machado","year":"2021","unstructured":"Machado PA, Gomes PS, Midlej V, Coimbra ES, de Matos Guedes HL. PF-429242, a subtilisin inhibitor, is effective in vitro against Leishmania infantum. Front Microbiol. 2021;12:583834. https:\/\/doi.org\/10.3389\/fmicb.2021.583834.","journal-title":"Front Microbiol"},{"issue":"7","key":"6630_CR23","doi-asserted-by":"publisher","first-page":"1373","DOI":"10.3390\/pharmaceutics14071373","volume":"14","author":"PA Machado","year":"2022","unstructured":"Machado PA, Gomes PS, Carneiro MPD, Midlej V, Coimbra ES, de Matos Guedes HL. Effects of a serine protease inhibitor N-p-tosyl-l-phenylalanine chloromethyl ketone (TPCK) on Leishmania amazonensis and Leishmania infantum. Pharmaceutics. 2022;14(7):1373. https:\/\/doi.org\/10.3390\/pharmaceutics14071373.","journal-title":"Pharmaceutics"},{"key":"6630_CR24","doi-asserted-by":"publisher","first-page":"1627","DOI":"10.1007\/s00436-007-0706-5","volume":"101","author":"RE Silva-Lopez","year":"2007","unstructured":"Silva-Lopez RE, Morgado-Diaz JA, Chavez MA, Giovanni-De-Simone S. Effects of serine protease inhibitors on viability and morphology of Leishmania (Leishmania) amazonensis promastigotes. Parasitol Res. 2007;101:1627\u201335. https:\/\/doi.org\/10.1007\/s00436-007-0706-5.","journal-title":"Parasitol Res"},{"key":"6630_CR25","first-page":"14","volume":"52","author":"T Chakraborti","year":"2015","unstructured":"Chakraborti T, Das P, Choudhury R, De T. Effect of different serine protease inhibitors in validating the 115 kDa Leishmania donovani secretory serine protease as chemotherapeutic target. Ind J Biochem Biophys. 2015;52:14\u201322.","journal-title":"Ind J Biochem Biophys"},{"key":"6630_CR26","doi-asserted-by":"publisher","first-page":"49","DOI":"10.1016\/j.molbiopara.2010.09.003","volume":"175","author":"JC Munday","year":"2011","unstructured":"Munday JC, McLuskey K, Brown E, Coombs GH, Mottram JC. Oligopeptidase B deficient mutants of Leishmania major. Mol Biochem Parasitol. 2011;175:49\u201357. https:\/\/doi.org\/10.1016\/j.molbiopara.2010.09.003.","journal-title":"Mol Biochem Parasitol"}],"container-title":["Parasites &amp; Vectors"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1186\/s13071-024-06630-w.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/article\/10.1186\/s13071-024-06630-w\/fulltext.html","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1186\/s13071-024-06630-w.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,1,23]],"date-time":"2025-01-23T18:57:30Z","timestamp":1737658650000},"score":1,"resource":{"primary":{"URL":"https:\/\/parasitesandvectors.biomedcentral.com\/articles\/10.1186\/s13071-024-06630-w"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,1,23]]},"references-count":26,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2025,12]]}},"alternative-id":["6630"],"URL":"https:\/\/doi.org\/10.1186\/s13071-024-06630-w","relation":{},"ISSN":["1756-3305"],"issn-type":[{"value":"1756-3305","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,1,23]]},"assertion":[{"value":"7 May 2024","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"12 December 2024","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"23 January 2025","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Declarations"}},{"value":"The study was conducted according to protocols approved by the Ethical Committee for Animal Handling (CEUA 080\/2018 from UFRJ).","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Ethics approval and consent to participate"}},{"value":"The authors declare no competing interests.","order":3,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"23"}}