{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,25]],"date-time":"2026-04-25T16:05:29Z","timestamp":1777133129108,"version":"3.51.4"},"reference-count":46,"publisher":"MDPI AG","issue":"11","license":[{"start":{"date-parts":[[2025,11,7]],"date-time":"2025-11-07T00:00:00Z","timestamp":1762473600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100003593","name":"Conselho Nacional de Desenvolvimento Cient\u00edfico e Tecnol\u00f3gico","doi-asserted-by":"publisher","award":["404927\/2023-8"],"award-info":[{"award-number":["404927\/2023-8"]}],"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":"crossref","id":[{"id":"10.13039\/501100002322","id-type":"DOI","asserted-by":"crossref"}]},{"name":"Federal University of Para\u00edba (UFPB), Brazil"},{"name":"FAPESQ-PB"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Antibiotics"],"abstract":"<jats:p>Background: Visceral leishmaniasis (VL) is a neglected tropical disease with limited therapeutic options, often restricted by toxicity, high costs, and resistance. Chalcones are promising scaffolds for the development of antiparasitic agents. Objectives: This study aimed to synthesize novel acetamides derived from 4-hydroxychalcones and evaluate their antileishmanial activity, cytotoxicity, potential synergy with amphotericin B (AmB), and mechanisms of action through in silico analyses. Methods: Six chalcone\u2013acetamides (3a\u2013c, 4a\u2013c) were synthesized and characterized by IR, NMR, and HRMS. In vitro activity against Leishmania infantum promastigotes and axenic amastigotes was assessed by colorimetric assays. Cytotoxicity was tested in human erythrocytes and PBMCs. Synergy with AmB was analyzed by the combination index. Molecular docking targeted parasite enzymes, and ADMET tools predicted pharmacokinetic and safety profiles. Results: Phenyl-substituted derivatives (3a\u2013c) were inactive, while cyclohexyl-substituted analogs (4a\u2013c) were active. Compound 4b displayed the strongest effect (IC50: 7.02 \u03bcM for promastigotes, 3.4 \u03bcM for amastigotes), with low cytotoxicity and high Selectivity Indices. In combination with AmB, compound 4b reduced the effective dose (DRI: 2.87) and increased the therapeutic window. Docking revealed favorable interactions of compound 4b with deubiquitinase DUB16 and tryparedoxin peroxidase I, suggesting enzyme inhibition. ADMET predictions supported good absorption and low toxicity. Conclusions: Compound 4b demonstrated potent and selective antileishmanial activity, synergism with AmB, and predicted safety. These findings highlight chalcone derivative 4b as a promising lead for future preclinical development in VL therapy.<\/jats:p>","DOI":"10.3390\/antibiotics14111123","type":"journal-article","created":{"date-parts":[[2025,11,7]],"date-time":"2025-11-07T09:14:45Z","timestamp":1762506885000},"page":"1123","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Novel Chalcone Derivatives as Anti-Leishmania infantum Agents with Potential Synergistic Activity and In Silico Insights"],"prefix":"10.3390","volume":"14","author":[{"ORCID":"https:\/\/orcid.org\/0009-0003-9753-7436","authenticated-orcid":false,"given":"Ana Let\u00edcia Monteiro","family":"Fernandes","sequence":"first","affiliation":[{"name":"Immunology Laboratory of Infectious Diseases (LABIDIC), Department of Cellular and Molecular Biology, Federal University of Paraiba, Jo\u00e3o Pessoa 58051-900, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0005-6585-2866","authenticated-orcid":false,"given":"Abra\u00e3o Pinheiro","family":"Sousa","sequence":"additional","affiliation":[{"name":"Bioenergy and Organic Synthesis Research Laboratory (LPBS), Department of Chemistry, Federal University of Para\u00edba, Jo\u00e3o Pessoa 58051-900, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6649-124X","authenticated-orcid":false,"given":"Delva Thyares Fonseca","family":"Lamec","sequence":"additional","affiliation":[{"name":"Immunology Laboratory of Infectious Diseases (LABIDIC), Department of Cellular and Molecular Biology, Federal University of Paraiba, Jo\u00e3o Pessoa 58051-900, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4924-0542","authenticated-orcid":false,"given":"Leonardo Lima","family":"Cardoso","sequence":"additional","affiliation":[{"name":"Immunology Laboratory of Infectious Diseases (LABIDIC), Department of Cellular and Molecular Biology, Federal University of Paraiba, Jo\u00e3o Pessoa 58051-900, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5360-6944","authenticated-orcid":false,"given":"Ros\u00e1lia Santos","family":"Ferreira","sequence":"additional","affiliation":[{"name":"Immunology Laboratory of Infectious Diseases (LABIDIC), Department of Cellular and Molecular Biology, Federal University of Paraiba, Jo\u00e3o Pessoa 58051-900, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8459-2717","authenticated-orcid":false,"given":"Shayenne Eduarda Ramos","family":"Vanderley","sequence":"additional","affiliation":[{"name":"Immunology Laboratory of Infectious Diseases (LABIDIC), Department of Cellular and Molecular Biology, Federal University of Paraiba, Jo\u00e3o Pessoa 58051-900, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0982-8261","authenticated-orcid":false,"given":"Petr\u00f4nio Filgueiras","family":"Athayde-Filho","sequence":"additional","affiliation":[{"name":"Bioenergy and Organic Synthesis Research Laboratory (LPBS), Department of Chemistry, Federal University of Para\u00edba, Jo\u00e3o Pessoa 58051-900, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8156-2272","authenticated-orcid":false,"given":"Gabriela Fehn","family":"Fiss","sequence":"additional","affiliation":[{"name":"Bioenergy and Organic Synthesis Research Laboratory (LPBS), Department of Chemistry, Federal University of Para\u00edba, Jo\u00e3o Pessoa 58051-900, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0964-389X","authenticated-orcid":false,"given":"Tatjana Souza Lima","family":"Keesen","sequence":"additional","affiliation":[{"name":"Immunology Laboratory of Infectious Diseases (LABIDIC), Department of Cellular and Molecular Biology, Federal University of Paraiba, Jo\u00e3o Pessoa 58051-900, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,11,7]]},"reference":[{"key":"ref_1","first-page":"2049936120966449","article-title":"Overcoming the global burden of neglected tropical diseases","volume":"7","year":"2020","journal-title":"Ther. Adv. Infect. Dis."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"622286","DOI":"10.3389\/fchem.2021.622286","article-title":"An overview on the therapeutics of neglected infectious diseases\u2014Leishmaniasis and Chagas diseases","volume":"9","author":"Brindha","year":"2021","journal-title":"Front. Chem."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"218","DOI":"10.3390\/immuno2010015","article-title":"Host\u2013pathogen interaction in leishmaniasis: Immune response and vaccination strategies","volume":"2","author":"Yasmin","year":"2022","journal-title":"Immuno"},{"key":"ref_4","unstructured":"Pan-American Health Organization (PAHO) (2025, November 03). Leishmaniasis: Epidemiological Report of the Americas Region. Available online: https:\/\/iris.paho.org\/handle\/10665.2\/51742."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"106661","DOI":"10.1016\/j.actatropica.2022.106661","article-title":"Amphotericin B: A drug of choice for Visceral Leishmaniasis","volume":"235","author":"Kumari","year":"2022","journal-title":"Acta Trop."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"660183","DOI":"10.3389\/fimmu.2021.660183","article-title":"Host-directed therapies for cutaneous leishmaniasis","volume":"12","author":"Novais","year":"2021","journal-title":"Front. Immunol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"758","DOI":"10.1016\/j.ejmech.2014.08.033","article-title":"Recent developments in biological activities of chalcones: A mini review","volume":"85","author":"Singh","year":"2014","journal-title":"Eur. J. Med. Chem."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"311","DOI":"10.1016\/j.ijantimicag.2017.06.010","article-title":"The chemotherapeutic potential of chalcones against leishmaniases: A review","volume":"51","author":"Tajuddeen","year":"2018","journal-title":"Int. J. Antimicrob. Agents"},{"key":"ref_9","first-page":"920","article-title":"A comprehensive review of chalcone derivatives as antileishmanial agents","volume":"150","author":"Domingos","year":"2019","journal-title":"Eur. J. Med. Chem."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1839","DOI":"10.1007\/s13738-021-02172-5","article-title":"Recent investigations into synthesis and pharmacological activities of phenoxy acetamide and its derivatives (chalcone, indole and quinoline) as possible therapeutic candidates","volume":"18","author":"Zabiulla","year":"2021","journal-title":"J. Iran. Chem. Soc."},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Santiago-Silva, K.M., Bortoleti, B.T.D.S., Oliveira, L.D.N., Maia, F.L.A., Castro, J.C., Costa, I.C., Lazarin, D.B., Wardell, J.L., Wardell, S.M.S.V., and Albuquerque, M.G. (2022). Antileishmanial Activity of 4,8-Dimethoxynaphthalenyl Chalcones on Leishmania amazonensis. Antibiotics, 11.","DOI":"10.3390\/antibiotics11101402"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"294","DOI":"10.1016\/j.cbi.2019.05.050","article-title":"Synergistic interactions of phytochemicals with antimicrobial agents: Potential strategy to counteract drug resistance","volume":"308","author":"Ayaz","year":"2019","journal-title":"Chem. Biol. Interact."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1044665","DOI":"10.3389\/fcimb.2022.1044665","article-title":"In vitro anti-Leishmania activity of triclabendazole and its synergic effect with amphotericin B","volume":"12","author":"Borges","year":"2023","journal-title":"Front. Cell. Infect. Microbiol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"e12813","DOI":"10.7717\/peerj.12813","article-title":"In vitro anti-Leishmania activity of 8-hydroxyquinoline and its synergistic effect with amphotericin B deoxycholate against Leishmania martiniquensis","volume":"10","author":"Chanmol","year":"2022","journal-title":"PeerJ"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"84","DOI":"10.15414\/jmbfs.2019.9.1.84-87","article-title":"Cyclohexane and its functionally substituted derivatives: Important class of organic compounds with potential antimicrobial activities","volume":"9","author":"Shoaib","year":"2019","journal-title":"J. Microbiol. Biotechnol. Food Sci."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"3342","DOI":"10.1016\/j.bmcl.2015.05.072","article-title":"Synthesis and evaluation of novel prenylated chalcone derivatives as anti-leishmanial and anti-trypanosomal compounds","volume":"25","author":"Passalacqua","year":"2015","journal-title":"Bioorg. Med. Chem. Lett."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Leite, F.F., Rodrigues, L.C., Oliveira, B.H.M.D., Duarte, G.D., Leite Ferreira, M.D., Sousa, N.F.D., Vanderley, S.E.R., Cardoso, L.L., Keesen, T.S.L., and Ara\u00fajo, R.S. (2024). In vitro and in silico evaluation of the anti-Leishmania activity of synthetic chalcones. Nat. Prod. Res., 1\u201312.","DOI":"10.1080\/14786419.2024.2401499"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"359","DOI":"10.1016\/j.ejmech.2014.05.034","article-title":"Synthesis and biological evaluation of chalcones as potential antileishmanial agents","volume":"81","author":"Gupta","year":"2014","journal-title":"Eur. J. Med. Chem."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"2300440","DOI":"10.1002\/ardp.202300440","article-title":"Synthesis and evaluation of hybrid sulfonamide-chalcones with potential antileishmanial activity","volume":"357","author":"Barreto","year":"2024","journal-title":"Arch. Pharm."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"2459","DOI":"10.1016\/j.bmcl.2017.04.010","article-title":"Computer-aided discovery of two novel chalcone-like compounds active and selective against Leishmania infantum","volume":"27","author":"Gomes","year":"2017","journal-title":"Bioorganic Med. Chem. Lett."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"2073","DOI":"10.1007\/s11030-022-10542-1","article-title":"Synthesis, characterization and evaluation of prenylated chalcones ethers as promising antileishmanial compounds","volume":"27","year":"2023","journal-title":"Mol. Divers."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"3342","DOI":"10.1021\/jm401893j","article-title":"Synthesis, structure\u2013activity relationships, and biological studies of chromenochalcones as potential antileishmanial agents","volume":"57","author":"Shivahare","year":"2014","journal-title":"J. Med. Chem."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1484","DOI":"10.1093\/bioinformatics\/btu841","article-title":"RRDistMaps: A UCSF Chimera tool for viewing and comparing protein distance maps","volume":"31","author":"Chen","year":"2015","journal-title":"Bioinformatics"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"8","DOI":"10.1002\/0471250953.bi0814s24","article-title":"Using AutoDock for ligand\u2013receptor docking","volume":"24","author":"Morris","year":"2008","journal-title":"Curr. Protoc. Bioinform."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"455","DOI":"10.1002\/jcc.21334","article-title":"AutoDock Vina: Improving the speed and accuracy of docking with a new scoring function, efficient optimization and multithreading","volume":"31","author":"Trott","year":"2010","journal-title":"J. Comput. Chem."},{"key":"ref_26","first-page":"82","article-title":"PyMOL: An open-source molecular graphics tool","volume":"40","author":"DeLano","year":"2002","journal-title":"CCP4 Newsl. Protein Crystallogr."},{"key":"ref_27","unstructured":"Dassault Syst\u00e8mes (2025, November 03). Discovery Studio Visualizer, Available online: https:\/\/www.3ds.com\/."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"4066","DOI":"10.1021\/acs.jmedchem.5b00104","article-title":"pkCSM: Predicting small-molecule pharmacokinetic and toxicity properties using graph-based signatures","volume":"58","author":"Pires","year":"2015","journal-title":"J. Med. Chem."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"42717","DOI":"10.1038\/srep42717","article-title":"SwissADME: A free web tool to evaluate pharmacokinetics, drug-likeness and medicinal chemistry friendliness of small molecules","volume":"7","author":"Daina","year":"2017","journal-title":"Sci. Rep."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"257","DOI":"10.1093\/nar\/gky318","article-title":"ProTox-II: A webserver for the prediction of toxicity of chemicals","volume":"46","author":"Banerjee","year":"2018","journal-title":"Nucleic Acids Res."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"985178","DOI":"10.3389\/fcimb.2022.985178","article-title":"The emerging role of deubiquitinases (DUBs) in parasites: A foresight review","volume":"12","author":"Kumar","year":"2022","journal-title":"Front. Cell. Infect. Microbiol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"e1008455","DOI":"10.1371\/journal.ppat.1008455","article-title":"Essential roles for deubiquitination in Leishmania life cycle progression","volume":"16","author":"Damianou","year":"2020","journal-title":"PLoS Pathog."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"e00081-22","DOI":"10.1128\/mcb.00081-22","article-title":"A novel role of secretory cytosolic tryparedoxin peroxidase in delaying apoptosis of Leishmania-infected macrophages","volume":"42","author":"Das","year":"2022","journal-title":"Mol. Cell. Biol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"3098","DOI":"10.1016\/j.bmcl.2016.05.013","article-title":"The synthesis and synergistic antifungal effects of chalcones against drug resistant Candida albicans","volume":"26","author":"Wang","year":"2016","journal-title":"Bioorg. Med. Chem. Lett."},{"key":"ref_35","first-page":"265","article-title":"Synergistic activity between conventional antifungals and chalcone-derived compound against dermatophyte fungi and Candida spp.","volume":"28","author":"Lemes","year":"2025","journal-title":"Int. Microbiol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"e13852728371713","DOI":"10.2174\/0113852728371713250529122641","article-title":"Novel 2-acetanilide 2-arylquinoline-4-carboxylates as antileishmanial agents: From prediction to in vitro activity\/toxicity","volume":"29","author":"Costa","year":"2025","journal-title":"Curr. Org. Chem."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"2897","DOI":"10.1055\/s-0034-1379209","article-title":"Homologation of isocyanates with lithium carbenoids: A straightforward access to \u03b1-halomethyl- and \u03b1,\u03b1-dihalomethylamides","volume":"46","author":"Pace","year":"2014","journal-title":"Synthesis"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"973","DOI":"10.1080\/00397911.2024.2356641","article-title":"Novel esters derived from 4-hydroxychalcones as potential sunscreens with antimicrobial action","volume":"54","author":"Sousa","year":"2024","journal-title":"Synth. Commun."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"184","DOI":"10.1107\/S2056989021000864","article-title":"Synthesis and structure of (E)-N-(4-methoxyphenyl)-2-[4-(3-oxo-3-phenylprop-1-en-1-yl)phenoxy]-acetamide","volume":"77","author":"Tien","year":"2021","journal-title":"Acta Cryst. E"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"205","DOI":"10.1016\/j.ijpara.2003.10.011","article-title":"Generation of Leishmania donovani axenic amastigotes: Their growth and biological characteristics","volume":"34","author":"Debrabant","year":"2004","journal-title":"Int. J. Parasitol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"795","DOI":"10.1016\/j.jconrel.2012.05.037","article-title":"PLGA nanoparticles and nanosuspensions with amphotericin B: Potent in vitro and in vivo alternatives to Fungizone and AmBisome","volume":"161","author":"Paulussen","year":"2012","journal-title":"J. Control. Release"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"235","DOI":"10.1093\/nar\/28.1.235","article-title":"The Protein Data Bank","volume":"28","author":"Berman","year":"2000","journal-title":"Nucleic Acids Res."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"969","DOI":"10.1042\/BCJ20253107","article-title":"Structure and activity of the essential UCH family deubiquitinase DUB16 from Leishmania donovani","volume":"482","author":"Brannigan","year":"2025","journal-title":"Biochem. J."},{"key":"ref_44","first-page":"1490","article-title":"The crystal structures of the tryparedoxin\u2013tryparedoxin peroxidase couple unveil the structural determinants of Leishmania detoxification pathway","volume":"21","author":"Fiorillo","year":"2012","journal-title":"Protein Sci."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"33","DOI":"10.1186\/1758-2946-3-33","article-title":"Open Babel: An open chemical toolbox","volume":"3","author":"Banck","year":"2011","journal-title":"J. Cheminform."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"440","DOI":"10.1158\/0008-5472.CAN-09-1947","article-title":"Drug combination studies and their synergy quantification using the Chou-Talalay method","volume":"70","author":"Chou","year":"2010","journal-title":"Cancer Res."}],"container-title":["Antibiotics"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2079-6382\/14\/11\/1123\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,11,9]],"date-time":"2025-11-09T05:12:21Z","timestamp":1762665141000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2079-6382\/14\/11\/1123"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,11,7]]},"references-count":46,"journal-issue":{"issue":"11","published-online":{"date-parts":[[2025,11]]}},"alternative-id":["antibiotics14111123"],"URL":"https:\/\/doi.org\/10.3390\/antibiotics14111123","relation":{},"ISSN":["2079-6382"],"issn-type":[{"value":"2079-6382","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,11,7]]}}}