{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,23]],"date-time":"2026-10-23T08:14:10Z","timestamp":1792743250645,"version":"4.5.0"},"reference-count":57,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2020,4,21]],"date-time":"2020-04-21T00:00:00Z","timestamp":1587427200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecules"],"abstract":"<jats:p>The protozoans Leishmania and Trypanosoma, belonging to the same Trypanosomatidae family, are the causative agents of Leishmaniasis, Chagas disease, and human African trypanosomiasis. Overall, these infections affect millions of people worldwide, posing a serious health issue as well as socio-economical concern. Current treatments are inadequate, mainly due to poor efficacy, toxicity, and emerging resistance; therefore, there is an urgent need for new drugs. Among several molecular targets proposed, trypanothione reductase (TR) is of particular interest for its critical role in controlling the parasite\u2019s redox homeostasis and several classes of active compounds that inhibit TR have been proposed so far. This review provides a comprehensive overview of TR\u2019s structural characterization. In particular, we discuss all the structural features of TR relevant for drug discovery, with a focus on the recent advances made in the understanding of inhibitor binding. The reported cases show how, on the basis of the detailed structural information provided by the crystallographic analysis, it is possible to rationally modify molecular scaffolds to improve their properties.<\/jats:p>","DOI":"10.3390\/molecules25081924","type":"journal-article","created":{"date-parts":[[2020,4,22]],"date-time":"2020-04-22T04:15:13Z","timestamp":1587528913000},"page":"1924","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":141,"title":["Targeting Trypanothione Reductase, a Key Enzyme in the Redox Trypanosomatid Metabolism, to Develop New Drugs against Leishmaniasis and Trypanosomiases"],"prefix":"10.3390","volume":"25","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-8401-890X","authenticated-orcid":false,"given":"Theo","family":"Battista","sequence":"first","affiliation":[{"name":"Department of Biochemical Sciences, Sapienza University, P.le A.Moro 5, 00185 Rome, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9913-0635","authenticated-orcid":false,"given":"Gianni","family":"Colotti","sequence":"additional","affiliation":[{"name":"Institute of Molecular Biology and Pathology, Italian National Research Council, IBPM-CNR, c\/o Department of Biochemical Sciences, Sapienza University, P.le A.Moro 5, 00185 Rome, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7754-399X","authenticated-orcid":false,"given":"Andrea","family":"Ilari","sequence":"additional","affiliation":[{"name":"Institute of Molecular Biology and Pathology, Italian National Research Council, IBPM-CNR, c\/o Department of Biochemical Sciences, Sapienza University, P.le A.Moro 5, 00185 Rome, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8966-8554","authenticated-orcid":false,"given":"Annarita","family":"Fiorillo","sequence":"additional","affiliation":[{"name":"Department of Biochemical Sciences, Sapienza University, P.le A.Moro 5, 00185 Rome, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2020,4,21]]},"reference":[{"key":"ref_1","unstructured":"World Health Organization (2020, March 30). WHO Publishes On-line Key Information about Leishmaniasis. Available online: https:\/\/www.who.int\/news-room\/fact-sheets\/detail\/leishmaniasis."},{"key":"ref_2","unstructured":"World Health Organization (2020, March 30). WHO Publishes On-line Key Information about Chagas Disease. Available online: https:\/\/www.who.int\/news-room\/fact-sheets\/detail\/chagas-disease-(american-trypanosomiasis)."},{"key":"ref_3","unstructured":"World Health Organization (2020, March 30). WHO Publishes On-line Key Information about HAT. Available online: https:\/\/www.who.int\/news-room\/fact-sheets\/detail\/trypanosomiasis-human-african-(sleeping-sickness)."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"217","DOI":"10.1038\/nrmicro.2016.193","article-title":"Anti-trypanosomatid drug discovery: An ongoing challenge and a continuing need","volume":"15","author":"Field","year":"2017","journal-title":"Nat. Rev. Microbiol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"71","DOI":"10.1098\/rstb.2008.0151","article-title":"Inflammation and oxidative stress in vertebrate host-parasite systems","volume":"364","author":"Sorci","year":"2009","journal-title":"Philos. Trans. R. Soc. Lond. B. Biol. Sci."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"121","DOI":"10.1016\/j.meegid.2016.06.054","article-title":"Catalase in Leishmaniinae: With me or against me?","volume":"50","author":"Kraeva","year":"2017","journal-title":"Infect. Genet. Evol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"409","DOI":"10.1126\/science.1112631","article-title":"The genome sequence of Trypanosoma cruzi, etiologic agent of Chagas disease","volume":"309","author":"Myler","year":"2005","journal-title":"Science"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"436","DOI":"10.1126\/science.1112680","article-title":"The genome of the kinetoplastid parasite, Leishmania major","volume":"309","author":"Ivens","year":"2005","journal-title":"Science"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"61","DOI":"10.4155\/fmc-2016-0180","article-title":"Polyamine-trypanothione pathway: An update","volume":"9","author":"Ilari","year":"2017","journal-title":"Future Med. Chem."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"589","DOI":"10.1016\/j.pt.2007.08.019","article-title":"Target assessment for antiparasitic drug discovery","volume":"23","author":"Frearson","year":"2007","journal-title":"Trends Parasitol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"5311","DOI":"10.1073\/pnas.95.9.5311","article-title":"Down-regulation of Leishmania donovani trypanothione reductase by heterologous expression of a trans-dominant mutant homologue: Effect on parasite intracellular survival","volume":"95","author":"Tovar","year":"1998","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"542","DOI":"10.1046\/j.1365-2958.2000.01721.x","article-title":"Trypanosomes lacking trypanothione reductase are avirulent and show increased sensitivity to oxidative stress","volume":"35","author":"Krieger","year":"2000","journal-title":"Mol. Microbiol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"460","DOI":"10.1111\/j.1432-1033.1995.tb20583.x","article-title":"Trypanothione reductase from Leishmania donovani. Purification, characterisation and inhibition by trivalent antimonials","volume":"230","author":"Cunningham","year":"1995","journal-title":"Eur. J. Biochem."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"2603","DOI":"10.1021\/jm900185q","article-title":"Molecular basis of antimony treatment in leishmaniasis","volume":"52","author":"Baiocco","year":"2009","journal-title":"J. Med. Chem."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"3069","DOI":"10.1021\/acs.molpharmaceut.8b00185","article-title":"Toward a Drug Against All Kinetoplastids: From LeishBox to Specific and Potent Trypanothione Reductase Inhibitors","volume":"15","author":"Ilari","year":"2018","journal-title":"Mol. Pharm."},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Beig, M., Oellien, F., Garoff, L., Noack, S., Krauth-Siegel, R.L., and Selzer, P.M. (2015). Trypanothione reductase: A target protein for a combined in vitro and in silico screening approach. PLoS Negl. Trop. Dis., 9.","DOI":"10.1371\/journal.pntd.0003773"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"2824","DOI":"10.1128\/AAC.01568-08","article-title":"Trypanothione reductase high-throughput screening campaign identifies novel classes of inhibitors with antiparasitic activity","volume":"53","author":"Holloway","year":"2009","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"5255","DOI":"10.1016\/j.bmcl.2011.07.036","article-title":"A fast virtual screening approach to identify structurally diverse inhibitors of trypanothione reductase","volume":"21","author":"Maccari","year":"2011","journal-title":"Bioorg. Med. Chem. Lett."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1280","DOI":"10.1016\/j.bmcl.2006.12.016","article-title":"High-throughput screening affords novel and selective trypanothione reductase inhibitors with anti-trypanosomal activity","volume":"17","author":"Martyn","year":"2007","journal-title":"Bioorg. Med. Chem. Lett."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1670","DOI":"10.1021\/jm801306g","article-title":"Development of a novel virtual screening cascade protocol to identify potential trypanothione reductase inhibitors","volume":"52","author":"Burgos","year":"2009","journal-title":"J. Med. Chem."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1333","DOI":"10.1002\/cmdc.200900097","article-title":"Improved tricyclic inhibitors of trypanothione reductase by screening and chemical synthesis","volume":"4","author":"Richardson","year":"2009","journal-title":"ChemMedChem"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"548","DOI":"10.1021\/jm001079l","article-title":"2- and 3-substituted 1,4-naphthoquinone derivatives as subversive substrates of trypanothione reductase and lipoamide dehydrogenase from Trypanosoma cruzi: Synthesis and correlation between redox cycling activities and in vitro cytotoxicity","volume":"44","author":"Buisine","year":"2001","journal-title":"J. Med. Chem."},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Turcano, L., Torrente, E., Missineo, A., Andreini, M., Gramiccia, M., Di Muccio, T., 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_24","doi-asserted-by":"crossref","unstructured":"Chac\u00f3n-Vargas, K.F., Nogueda-Torres, B., S\u00e1nchez-Torres, L.E., Suarez-Contreras, E., Villalobos-Rocha, J.C., Torres-Martinez, Y., Lara-Ramirez, E.E., Fiorani, G., Krauth-Siegel, R.L., and Bolognesi, M.L. (2017). Trypanocidal Activity of Quinoxaline 1,4 Di-N-oxide Derivatives as Trypanothione Reductase Inhibitors. Molecules, 22.","DOI":"10.3390\/molecules22020220"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"247","DOI":"10.1007\/s00726-019-02731-4","article-title":"Structure-guided approach to identify a novel class of anti-leishmaniasis diaryl sulfide compounds targeting the trypanothione metabolism","volume":"52","author":"Colotti","year":"2019","journal-title":"Amino Acids"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"36","DOI":"10.1016\/j.compbiolchem.2019.01.009","article-title":"In silico identification and evaluation of new Trypanosoma cruzi trypanothione reductase (TcTR) inhibitors obtained from natural products database of the Bahia semi-arid region (NatProDB)","volume":"79","author":"Pita","year":"2019","journal-title":"Comput. Biol. Chem."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"11416","DOI":"10.1002\/chem.201901664","article-title":"Targeting a Large Active Site: Structure-Based Design of Nanomolar Inhibitors of Trypanosoma brucei Trypanothione Reductase","volume":"25","author":"Halgas","year":"2019","journal-title":"Chemistry"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"340","DOI":"10.1016\/j.ejmech.2016.08.048","article-title":"Synthesis and evaluation against Leishmania amazonensis of novel pyrazolo[3,4-d]pyridazinone-N-acylhydrazone-(bi)thiophene hybrids","volume":"124","author":"Jacomini","year":"2016","journal-title":"Eur. J. Med. Chem."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"655","DOI":"10.1016\/j.ejmech.2018.02.087","article-title":"Synthesis and antikinetoplastid evaluation of bis(benzyl)spermidine derivatives","volume":"150","author":"Jagu","year":"2018","journal-title":"Eur. J. Med. Chem."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"527","DOI":"10.1016\/j.ejmech.2018.04.057","article-title":"Identification of chalcone-based antileishmanial agents targeting trypanothione reductase","volume":"152","author":"Ortalli","year":"2018","journal-title":"Eur. J. Med. Chem."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"141","DOI":"10.1080\/07391102.2015.1135298","article-title":"Febrifugine analogues as Leishmania donovani trypanothione reductase inhibitors: Binding energy analysis assisted by molecular docking, ADMET and molecular dynamics simulation","volume":"35","author":"Pandey","year":"2017","journal-title":"J. Biomol. Struct. Dyn."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"304","DOI":"10.1080\/14756366.2016.1250755","article-title":"Inhibition of Leishmania infantum trypanothione reductase by diaryl sulfide derivatives","volume":"32","author":"Saccoliti","year":"2017","journal-title":"J. Enzyme. Inhib. Med. Chem."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"4850","DOI":"10.1016\/j.bmc.2018.08.025","article-title":"Synthesis and SAR of new isoxazole-triazole bis-heterocyclic compounds as analogues of natural lignans with antiparasitic activity","volume":"26","author":"Zimmermann","year":"2018","journal-title":"Bioorg. Med. Chem."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"67","DOI":"10.1111\/j.1432-1033.1993.tb17734.x","article-title":"Substrate interactions between trypanothione reductase and N1-glutathionylspermidine disulphide at 0.28-nm resolution","volume":"213","author":"Bailey","year":"1993","journal-title":"Eur. J. Biochem."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1175","DOI":"10.1002\/cmdc.201300176","article-title":"Inhibition of Leishmania infantum trypanothione reductase by azole-based compounds: A comparative analysis with its physiological substrate by X-ray crystallography","volume":"8","author":"Baiocco","year":"2013","journal-title":"ChemMedChem"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"12","DOI":"10.1016\/j.molbiopara.2009.09.002","article-title":"Comparative structural, kinetic and inhibitor studies of Trypanosoma brucei trypanothione reductase with T. cruzi","volume":"169","author":"Jones","year":"2010","journal-title":"Mol. Biochem. Parasitol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"6514","DOI":"10.1021\/jm200312v","article-title":"Dihydroquinazolines as a novel class of Trypanosoma brucei trypanothione reductase inhibitors: Discovery, synthesis, and characterization of their binding mode by protein crystallography","volume":"54","author":"Patterson","year":"2011","journal-title":"J. Med. Chem."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"695","DOI":"10.1146\/annurev.mi.46.100192.003403","article-title":"Metabolism and functions of trypanothione in the Kinetoplastida","volume":"46","author":"Fairlamb","year":"1992","journal-title":"Annu. Rev. Microbiol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"6437","DOI":"10.1021\/bi963074p","article-title":"Glutathione reductase turned into trypanothione reductase: Structural analysis of an engineered change in substrate specificity","volume":"36","author":"Stoll","year":"1997","journal-title":"Biochemistry"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"1880","DOI":"10.1002\/cmdc.201402032","article-title":"Binding to large enzyme pockets: Small-molecule inhibitors of trypanothione reductase","volume":"9","author":"Persch","year":"2014","journal-title":"ChemMedChem"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"873","DOI":"10.1021\/acsinfecdis.8b00355","article-title":"Pyrrolopyrimidine vs. Imidazole-Phenyl-Thiazole Scaffolds in Nonpeptidic Dimerization Inhibitors of Leishmania infantum Trypanothione Reductase","volume":"5","author":"Revuelto","year":"2019","journal-title":"ACS Infect. Dis."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"73","DOI":"10.1002\/(SICI)1097-0134(199601)24:1<73::AID-PROT5>3.0.CO;2-P","article-title":"Crystal structure of the Trypanosoma cruzi trypanothione reductase.mepacrine complex","volume":"24","author":"Jacoby","year":"1996","journal-title":"Proteins"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"29493","DOI":"10.1074\/jbc.M403187200","article-title":"Two interacting binding sites for quinacrine derivatives in the active site of trypanothione reductase: A template for drug design","volume":"279","author":"Saravanamuthu","year":"2004","journal-title":"J. Biol. Chem."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"1341","DOI":"10.1002\/cmdc.200900098","article-title":"Synthesis and evaluation of 1-(1-(Benzo[b]thiophen-2-yl)cyclohexyl) piperidine (BTCP) analogues as inhibitors of trypanothione reductase","volume":"4","author":"Patterson","year":"2009","journal-title":"ChemMedChem"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"292","DOI":"10.1002\/cmdc.201000420","article-title":"Improved inhibitors of trypanothione reductase by combination of motifs: Synthesis, inhibitory potency, binding mode, and antiprotozoal activities","volume":"6","author":"Eberle","year":"2011","journal-title":"ChemMedChem"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"957","DOI":"10.1002\/cmdc.201800067","article-title":"Biological Evaluation and X-ray Co-crystal Structures of Cyclohexylpyrrolidine Ligands for Trypanothione Reductase, an Enzyme from the Redox Metabolism of Trypanosoma","volume":"13","author":"Bryson","year":"2018","journal-title":"ChemMedChem"},{"key":"ref_47","doi-asserted-by":"crossref","unstructured":"Turcano, L.B.T., Torrente De Haro, E., Missineo, A., Alli, C., Paonessa, G., Colotti, G., Harper, S.F.A., Ilari, A., and Bresciani, A. (2020). Spiro-containing derivateives show antiparasitic activity against Trypanosoma brucei through inhibition of the trypanothione reductase enzyme. PLoS Negl. Trop. Dis., accepted.","DOI":"10.1371\/journal.pntd.0008339"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"467","DOI":"10.1111\/j.1476-5381.1987.tb11196.x","article-title":"Pharmacological characterization of binding sites identified in rat brain following in vivo administration of [3H]-spiperone","volume":"90","author":"Chivers","year":"1987","journal-title":"Br. J. Pharmacol."},{"key":"ref_49","first-page":"1689","article-title":"The pharmacology of fluspirilene (R 6218), a potent, long-acting and injectable neuroleptic drug","volume":"20","author":"Janssen","year":"1970","journal-title":"Arzneimittelforschung"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"3935","DOI":"10.1039\/b806371k","article-title":"Diaryl sulfide-based inhibitors of trypanothione reductase: Inhibition potency, revised binding mode and antiprotozoal activities","volume":"6","author":"Stump","year":"2008","journal-title":"Org. Biomol. Chem."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"2034","DOI":"10.1002\/cmdc.200900327","article-title":"Synthesis, inhibition potency, binding mode, and antiprotozoal activities of fluorescent inhibitors of trypanothione reductase based on mepacrine-conjugated diaryl sulfide scaffolds","volume":"4","author":"Eberle","year":"2009","journal-title":"ChemMedChem"},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"954","DOI":"10.2741\/4628","article-title":"Metal- and metalloid-containing drugs for the treatment of trypanosomatid diseases","volume":"23","author":"Colotti","year":"2018","journal-title":"Front. Biosci."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"230","DOI":"10.1021\/ml1002629","article-title":"Inhibitory Effect of Silver Nanoparticles on Trypanothione Reductase Activity and Leishmania infantum Proliferation","volume":"2","author":"Baiocco","year":"2011","journal-title":"ACS Med. Chem. Lett."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"803","DOI":"10.1007\/s00726-011-0997-9","article-title":"A gold-containing drug against parasitic polyamine metabolism: The X-ray structure of trypanothione reductase from Leishmania infantum in complex with auranofin reveals a dual mechanism of enzyme inhibition","volume":"42","author":"Ilari","year":"2012","journal-title":"Amino Acids"},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"1634","DOI":"10.1002\/cmdc.201300276","article-title":"Metal-based compounds as prospective antileishmanial agents: Inhibition of trypanothione reductase by selected gold complexes","volume":"8","author":"Colotti","year":"2013","journal-title":"ChemMedChem"},{"key":"ref_56","unstructured":"Drugbank (2020, April 14). Auranofin. Available online: https:\/\/www.drugbank.ca\/drugs\/DB00995."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"1212","DOI":"10.1002\/cbic.201200744","article-title":"Probing the dimerization interface of Leishmania infantum trypanothione reductase with site-directed mutagenesis and short peptides","volume":"14","author":"Toro","year":"2013","journal-title":"Chembiochem"}],"container-title":["Molecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1420-3049\/25\/8\/1924\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,13]],"date-time":"2025-10-13T13:21:35Z","timestamp":1760361695000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1420-3049\/25\/8\/1924"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,4,21]]},"references-count":57,"journal-issue":{"issue":"8","published-online":{"date-parts":[[2020,4]]}},"alternative-id":["molecules25081924"],"URL":"https:\/\/doi.org\/10.3390\/molecules25081924","relation":{},"ISSN":["1420-3049"],"issn-type":[{"value":"1420-3049","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,4,21]]}}}