{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,10]],"date-time":"2026-02-10T04:43:08Z","timestamp":1770698588797,"version":"3.49.0"},"reference-count":37,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2026,2,9]],"date-time":"2026-02-09T00:00:00Z","timestamp":1770595200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Microorganisms"],"abstract":"<jats:p>Leishmaniasis remains a neglected tropical disease with treatment limitations driven by toxicity, cost, and emerging resistance. Trypanothione reductase (TryR) and glycerol-3-phosphate dehydrogenase (GPDH) are essential Leishmania enzymes supporting redox homeostasis and energy\/redox-linked metabolism, providing a biologically grounded rationale for dual-target inhibition. We applied an integrated in silico workflow to prioritize candidate inhibitors using ADMET prediction (SwissADME\/pkCSM), molecular docking (AutoDock Vina), and 100 ns molecular dynamics (MD) simulations; human GPDH was included as a negative control to probe potential off-target liability. ADMET screening identified 41 drug-like candidates, with most predicted to have high GI absorption and low toxicity flags across assessed endpoints (computational predictions interpreted cautiously). Docking highlighted two leading compounds. CID 6529858 showed the most favorable predicted binding to Leishmania GPDH (\u22128.9 kcal\/mol) with a modest parasite-favored score difference versus human GPDH (\u22127.2 kcal\/mol; \u0394 = \u22121.7 kcal\/mol), while eupatorin (CID: 97214) displayed dual-target potential (TryR \u22127.5 kcal\/mol; Leishmania GPDH \u22128.2 kcal\/mol; human GPDH \u22127.2 kcal\/mol; \u0394 = \u22121.0 kcal\/mol). In MD, key complexes remained stable: CID 6529858 exhibited low GPDH backbone deviation (~0.25\u20130.40 nm), and eupatorin showed the most stable TryR trajectory (average RMSD ~0.45 nm), supported by generally low residue fluctuations across complexes. PCA further suggested ligand-associated restriction of large-scale motions (e.g., GPDH PC1 = 27.38%; TryR PC1 = 18.1%). Overall, these results nominate eupatorin as a promising dual-target lead and CID 6529858 as a strong GPDH-focused scaffold, warranting experimental enzyme inhibition, antiparasitic efficacy, and host\u2013cell cytotoxicity testing to confirm potency and selectivity.<\/jats:p>","DOI":"10.3390\/microorganisms14020407","type":"journal-article","created":{"date-parts":[[2026,2,9]],"date-time":"2026-02-09T10:45:31Z","timestamp":1770633931000},"page":"407","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["In Silico Targeting of Trypanothione Reductase and Glycerol-3-Phosphate Dehydrogenase in Leishmania"],"prefix":"10.3390","volume":"14","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1463-4488","authenticated-orcid":false,"given":"Ali","family":"Alisaac","sequence":"first","affiliation":[{"name":"Faculty of Applied Medical Sciences, Al-Baha University, Al-Baha 65779, Saudi Arabia"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2026,2,9]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"121","DOI":"10.1007\/s40475-021-00232-7","article-title":"A Review of Leishmaniasis: Current Knowledge and Future Directions","volume":"8","author":"Mann","year":"2021","journal-title":"Curr. Trop. Med. Rep."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"11305","DOI":"10.1021\/cr500365f","article-title":"Recent Developments in Drug Discovery for Leishmaniasis and Human African Trypanosomiasis","volume":"114","author":"Nagle","year":"2014","journal-title":"Chem. Rev."},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Zhang, H., Yan, R., Liu, Y., Yu, M., He, Z., Xiao, J., Li, K., Liu, G., Ning, Q., and Li, Y. (2025). Progress in Antileishmanial Drugs: Mechanisms, Challenges, and Prospects. PLoS Negl. Trop. Dis., 19.","DOI":"10.1371\/journal.pntd.0012735"},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Mendoza-Le\u00f3n, A., Serrano, G., and Ponte-Sucre, A. (2023). Challenges in Drug Discovery and Description Targeting Leishmania spp.: Enzymes, Structural Proteins, and Transporters. Front. Trop. Dis., 4.","DOI":"10.3389\/fitd.2023.1241044"},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Gonz\u00e1lez-Montero, M.C., Andr\u00e9s-Rodr\u00edguez, J., Garc\u00eda-Fern\u00e1ndez, N., P\u00e9rez-Pertejo, Y., Reguera, R.M., Bala\u00f1a-Fouce, R., and Garc\u00eda-Estrada, C. (2024). Targeting Trypanothione Metabolism in Trypanosomatids. Molecules, 29.","DOI":"10.3390\/molecules29102214"},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Valli\u00e8res, C., Golinelli-Cohen, M.P., Guittet, O., Lepoivre, M., Huang, M.E., and Vernis, L. (2023). Redox-Based Strategies Against Infections by Eukaryotic Pathogens. Genes, 14.","DOI":"10.3390\/genes14040778"},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Lazarev, V.F., Guzhova, I.V., and Margulis, B.A. (2020). Glyceraldehyde-3-Phosphate Dehydrogenase Is a Multifaceted Therapeutic Target. Pharmaceutics, 12.","DOI":"10.3390\/pharmaceutics12050416"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1055","DOI":"10.1002\/med.22005","article-title":"Experimental Structure-Based Drug Design (SBDD) Applications for Anti-Leishmanial Drugs: A Paradigm Shift?","volume":"44","year":"2024","journal-title":"Med. Res. Rev."},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Turcano, L., Torrente, E., Missineo, A., Andreini, M., Gramiccia, M., Muccio, T.D., Genovese, I., Fiorillo, A., Harper, S., and Bresciani, A. (2018). Identification and Binding Mode of a Novel Leishmania Trypanothione Reductase Inhibitor from High Throughput Screening. PLoS Negl. Trop. Dis., 12.","DOI":"10.1371\/journal.pntd.0006969"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"2476","DOI":"10.1016\/j.bmc.2009.01.079","article-title":"Discovery of Novel Trypanosoma cruzi Glyceraldehyde-3-Phosphate Dehydrogenase Inhibitors","volume":"17","author":"Freitas","year":"2009","journal-title":"Bioorg. Med. Chem."},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Rabaan, A.A., Bakhrebah, M.A., Mohapatra, R.K., Farahat, R.A., Dhawan, M., Alwarthan, S., Aljeldah, M., Al Shammari, B.R., Al-Najjar, A.H., and Alhusayyen, M.A. (2023). Omics approaches in drug development against leishmaniasis: Current scenario and future prospects. Pathogens, 12.","DOI":"10.3390\/pathogens12010039"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"109","DOI":"10.1016\/j.ejmech.2015.10.037","article-title":"Comparative study of structural models of Leishmania donovani and human GDP-mannose pyrophosphorylases","volume":"107","author":"Daligaux","year":"2016","journal-title":"Eur. J. Med. Chem."},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"dos Santos Vasconcelos, C.R., and Rezende, A.M. (2021). Systematic in silico evaluation of Leishmania spp. proteomes for drug discovery. Front. Chem., 9.","DOI":"10.3389\/fchem.2021.607139"},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Gervazoni, L.F.O., Barcellos, G.B., Ferreira-Paes, T., and Almeida-Amaral, E.E. (2020). Use of natural products in leishmaniasis chemotherapy: An overview. Front. Chem., 8.","DOI":"10.3389\/fchem.2020.579891"},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Singh, R., Kashif, M., Srivastava, P., and Manna, P.P. (2023). Recent advances in chemotherapeutics for leishmaniasis: Importance of the cellular biochemistry of the parasite and its molecular interaction with the host. Pathogens, 12.","DOI":"10.3390\/pathogens12050706"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"249","DOI":"10.1016\/j.drudis.2016.11.001","article-title":"Molecular dynamics-driven drug discovery: Leaping forward with confidence","volume":"22","author":"Ganesan","year":"2017","journal-title":"Drug Discov. Today"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"334","DOI":"10.1124\/pr.112.007336","article-title":"Computational methods in drug discovery","volume":"66","author":"Sliwoski","year":"2014","journal-title":"Pharmacol. Rev."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1189","DOI":"10.1016\/S1074-5521(02)00243-0","article-title":"Anomalous differences of light elements in determining precise binding modes of ligands to glycerol-3-phosphate dehydrogenase","volume":"9","author":"Choe","year":"2002","journal-title":"Chem. Biol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"11253","DOI":"10.1021\/acscatal.0c02757","article-title":"Modeling the role of a flexible loop and active site side chains in hydride transfer catalyzed by glycerol-3-phosphate dehydrogenase","volume":"10","author":"Mhashal","year":"2020","journal-title":"ACS Catal."},{"key":"ref_20","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_21","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_22","unstructured":"Hempel, J.E., Williams, C.H., and Hong, C.C. (2015). Small-molecule library screening by docking with PyRx. Chemical Biology: Methods and Protocols, Springer."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"459","DOI":"10.1021\/ct900549r","article-title":"Implementation of the CHARMM force field in GROMACS: Analysis of protein stability effects from correction maps, virtual interaction sites, and water models","volume":"6","author":"Bjelkmar","year":"2010","journal-title":"J. Chem. Theory Comput."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Salo-Ahen, O.M.H., Alanko, I., Bhadane, R., Bonvin, A.M.J.J., Honorato, R.V., Hossain, S., Juffer, A.H., Kabedev, A., Lahtela-Kakkonen, M., and Larsen, A.S. (2021). Molecular dynamics simulations in drug discovery and pharmaceutical development. Processes, 9.","DOI":"10.3390\/pr9010071"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"738","DOI":"10.1016\/j.coph.2010.09.016","article-title":"Molecular dynamics simulations of protein dynamics and their relevance to drug discovery","volume":"10","author":"Salsbury","year":"2010","journal-title":"Curr. Opin. Pharmacol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"120829","DOI":"10.1016\/j.lfs.2022.120829","article-title":"Virulence factors of Leishmania parasite: Their paramount importance in unraveling novel vaccine candidates and therapeutic targets","volume":"306","author":"Kumari","year":"2022","journal-title":"Life Sci."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"626838","DOI":"10.1155\/2012\/626838","article-title":"Developments in diagnosis and antileishmanial drugs","volume":"2012","author":"Bhargava","year":"2012","journal-title":"Interdiscip. Perspect. Infect. Dis."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"306","DOI":"10.1021\/acsomega.4c06146","article-title":"In silico structural insights and potential inhibitor identification based on the benzothiazole core for targeting Leishmania major pteridine reductase 1","volume":"10","author":"Weber","year":"2025","journal-title":"ACS Omega"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"464","DOI":"10.1017\/S0031182016002523","article-title":"Chemotherapy of leishmaniasis: Present challenges","volume":"145","author":"Uliana","year":"2018","journal-title":"Parasitology"},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Ponte-Sucre, A., Gamarro, F., Dujardin, J.-C., Barrett, M.P., L\u00f3pez-V\u00e9lez, R., Garc\u00eda-Hern\u00e1ndez, R., Pountain, A.W., Mwenechanya, R., and Papadopoulou, B. (2017). Drug resistance and treatment failure in leishmaniasis: A 21st century challenge. PLoS Negl. Trop. Dis., 11.","DOI":"10.1371\/journal.pntd.0006052"},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Wijnant, G.-J., Dumetz, F., Dirkx, L., Bult\u00e9, D., Cuypers, B., Van Bocxlaer, K., and Hendrickx, S. (2022). Tackling Drug Resistance and Other Causes of Treatment Failure in Leishmaniasis. Front Trop Dis., 3.","DOI":"10.3389\/fitd.2022.837460"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"e180377","DOI":"10.1590\/0074-02760180377","article-title":"Ascorbate peroxidase overexpression protects Leishmania braziliensis against trivalent antimony effects","volume":"113","author":"Xavier","year":"2018","journal-title":"Mem\u00f3rias Inst. Oswaldo Cruz"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"350","DOI":"10.1016\/j.redox.2017.03.004","article-title":"Deciphering the interplay between cysteine synthase and thiol cascade proteins in modulating Amphotericin B resistance and survival of Leishmania donovani under oxidative stress","volume":"12","author":"Singh","year":"2017","journal-title":"Redox Biol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"e01766-19","DOI":"10.1128\/AAC.01766-19","article-title":"Leishmanicidal Activity of an In Silico-Screened Novel Inhibitor against Ascorbate Peroxidase of Leishmania donovani","volume":"64","author":"Kashif","year":"2020","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Matadamas-Mart\u00ednez, F., Hern\u00e1ndez-Campos, A., T\u00e9llez-Valencia, A., V\u00e1zquez-Raygoza, A., Compar\u00e1n-Alarc\u00f3n, S., Y\u00e9pez-Mulia, L., and Castillo, R. (2019). Leishmania mexicana Trypanothione Reductase Inhibitors: Computational and Biological Studies. Molecules, 24.","DOI":"10.3390\/molecules24183216"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"2025","DOI":"10.1007\/s00436-020-06736-x","article-title":"An overview of biochemically characterized drug targets in metabolic pathways of Leishmania parasite","volume":"119","author":"Raj","year":"2020","journal-title":"Parasitol. Res."},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Al-Khalaifah, H.S. (2022). Major Molecular Factors Related to Leishmania Pathogenicity. Front. Immunol., 13.","DOI":"10.3389\/fimmu.2022.847797"}],"container-title":["Microorganisms"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-2607\/14\/2\/407\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,2,9]],"date-time":"2026-02-09T11:06:03Z","timestamp":1770635163000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-2607\/14\/2\/407"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,2,9]]},"references-count":37,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2026,2]]}},"alternative-id":["microorganisms14020407"],"URL":"https:\/\/doi.org\/10.3390\/microorganisms14020407","relation":{},"ISSN":["2076-2607"],"issn-type":[{"value":"2076-2607","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,2,9]]}}}