[en] Trypanosomiasis is a parasitic disease for which new treatments are needed due to the frequent occurrence of adverse side effects of current available drugs. Natural compounds found in traditionally used plants offer opportunities to discover innovative compounds that could prove pivotal to antitrypanosomal drug development. 3-O-p-(Z/E)-coumaroyltormentic acids (CTA) were isolated first from the West Africa-native tree Vitellaria paradoxa and have demonstrated quite selective in vitro and in vivo antitrypanosomal activity, despite the unknown mode of action. In this study, a metabolomics analysis using the data from both LC-HR-MS and 1H-NMR described CTA's effects on Trypanosoma brucei after 3 h exposure under 5 or 10 x EC50. Our study shows CTA's activity impacted tryptophan metabolism and reveals potential targets in different branches of this metabolism. Our results demonstrate a likely presence of enzymes dedicated to tryptophan, like a tryptophan aminotransferase, tryptophan 2,3-dioxygenase and/or indoleamine 2,3-dioxygenase, and other enzymes of the kynurenine pathway, despite the absence of their description thus far in this species. These data further implicate that CTA's toxic effect on the tryptophan metabolism may be attributed to the decrease of the intracellular level of essential aspartate, resulting from inhibition of its aminotransferase. In resume, our study shines light on the likelihood of the tryptophan metabolism pathway presenting innovative targets toward the development of antitrypanosomal drugs. These require confirmation through functional and enzymatic studies.
Disciplines :
Pharmacy, pharmacology & toxicology
Author, co-author :
Mamede, Lúcia ; Pharmacognosy Research Group, Louvain Drug Research Institute (LDRI), UCLouvain, Brussels, Belgium, Laboratory of Pharmacognosy, Center of Interdisciplinary Research on Medicines (CIRM), University of Liège, Belgium. Electronic address: phd.lucia.mamede@outlook.com
Fall, Fanta; Pharmacognosy Research Group, Louvain Drug Research Institute (LDRI), UCLouvain, Brussels, Belgium
Vast, Madeline; Institute of Statistics, Biostatistics and Actuarial Sciences (ISBA/LIDAM), Université catholique de Louvain (UCLouvain), Louvain-la-Neuve, Belgium
Hughes, Kristelle ; Pharmacognosy Research Group, Louvain Drug Research Institute (LDRI), UCLouvain, Brussels, Belgium
Martelli, Giorgia ; Pharmacognosy Research Group, Louvain Drug Research Institute (LDRI), UCLouvain, Brussels, Belgium
Caligiore, Francesco ; de Duve Institute and WELBIO, UCLouvain, Avenue Hippocrate 75, 1200, Brussels, Belgium
Govaerts, Bernadette; Institute of Statistics, Biostatistics and Actuarial Sciences (ISBA/LIDAM), Université catholique de Louvain (UCLouvain), Louvain-la-Neuve, Belgium
Michels, Paul A M; School of Biological Sciences, The University of Edinburgh, Scotland, UK
Frederich, Michel ; Université de Liège - ULiège > Département de pharmacie > Pharmacognosie
Quetin-Leclercq, Joëlle; Pharmacognosy Research Group, Louvain Drug Research Institute (LDRI), UCLouvain, Brussels, Belgium
Language :
English
Title :
Metabolomics study of 3-O-p-(Z/E)-coumaroyltormentic acid-treated Trypanosoma brucei brucei.
Publication date :
August 2025
Journal title :
International Journal for Parasitology: Drugs and Drug Resistance
The authors would like to thank Sylvain Legay for in vitro-cultivated apple cells. This work was supported by the Belgian National Fund for Scientific Research (FNRS , grant PDR T.0092.20 ).
Adusumilli, R., Mallick, P., Data conversion with ProteoWizard msConvert. Methods Mol. Biol. 1550 (2017), 339–368, 10.1007/978-1-4939-6747-6_23.
Afolabi, J.O., Olorode, E.M., Olomola, D.B., Fasakin, Y.O., Adekunle, E.A., Effects of different media strengths and hormone concentrations on in-vitro regeneration of Vitellaria paradoxa C.F. Gaertn. Nigerian J. Biotechnol. 37:1 (2020), 150–158, 10.4314/njb.v37i1.16.
Barrett, M.P., Priotto, G., Franco, J.R., Lejon, V., Lindner, A.K., Elimination of human African trypanosomiasis: the long last mile. PLoS Neglected Trop. Dis., 18(5), 2024, e0012091, 10.1371/journal.pntd.0012091.
Biebinger, S., Elizabeth Wirtz, L., Lorenz, P., Christine, Clayton, Vectors for inducible expression of toxic gene products in bloodstream and procyclic Trypanosoma brucei. Mol. Biochem. Parasitol. 85:1 (1997), 99–112, 10.1016/S0166-6851(96)02815-0.
Catteau, L., Schioppa, L., Beaufay, C., Girardi, C., Hérent, M.-F., Frédérich, M., Quetin-Leclercq, J., Antiprotozoal activities of triterpenic acids and ester derivatives isolated from the leaves of Vitellaria paradoxa. Planta Med. 87:10–11 (2021), 860–867, 10.1055/a-1286-1879.
Chong, J., Wishart, D.S., Xia, J., Using MetaboAnalyst 4.0 for comprehensive and integrative metabolomics data analysis. Curr. Protocol. Bioinf. 68:1 (2019), 1–128, 10.1002/cpbi.86.
Creek, D.J., Nijagal, B., Kim, D.-H., Rojas, F., Matthews, K.R., Barrett, M.P., Metabolomics guides rational development of a simplified cell culture medium for drug screening against trypanosoma brucei. Antimicrob. Agents Chemother. 57:6 (2013), 2768–2779, 10.1128/AAC.00044-13.
Diskin, C., Corcoran, S.E., Tyrrell, V.J., McGettrick, A.F., Zaslona, Z., O'Donnell, V.B., et al. The trypanosome-derived metabolite indole-3-pyruvate inhibits prostaglandin production in macrophages by targeting COX2. J. Immunol. 207:10 (2021), 2551–2560, 10.4049/jimmunol.2100402.
Ecco, G., Vernal, J., Razzera, G., Tavares, C., Serpa, V.I., Arias, S., et al. Initial characterization of a recombinant kynureninase from Trypanosoma cruzi identified from an EST database. Gene 448:1 (2009), 1–6, 10.1016/j.gene.2009.08.007.
EL Sawalhy, A., Seed, J.R., EL Attar, H., Hall, J.E., Catabolism of tryptophan by trypanosoma evansi. J. Eukaryot. Microbiol. 42:6 (1995), 684–690, 10.1111/j.1550-7408.1995.tb01616.x.
Fall, F., Desmet, L., Mamede, L., Schioppa, L., De Tullio, P., Frédérich, M., et al. Comparison of three widely employed extraction methods for metabolomic analysis of Trypanosoma brucei. Curr. Protocol., 4(5), 2024, e1043, 10.1002/cpz1.1043.
Fall, F., Mamede, L., Schioppa, L., Ledoux, A., De Tullio, P., Michels, P., et al. Trypanosoma brucei: metabolomics for analysis of cellular metabolism and drug discovery. Metabolomics, 18(4), 2022, 20, 10.1007/s11306-022-01880-0.
Féraud, B., Munaut, C., Martin, M., Verleysen, M., Govaerts, B., Combining strong sparsity and competitive predictive power with the L-sOPLS approach for biomarker discovery in metabolomics. Metabolomics 13:11 (2017), 1–15, 10.1007/s11306-017-1275-y.
Franco, J., Scarone, L., Comini, M.A., Chapter three - drugs and drug resistance in african and American trypanosomiasis. Botta, M., (eds.) Annual Reports in Medicinal Chemistry, vol. 51, 2018, Academic Press, 97–133, 10.1016/bs.armc.2018.08.003.
Giacomoni, F., Le Corguillé, G., Monsoor, M., Landi, M., Pericard, P., Pétéra, M., et al. Workflow4Metabolomics: a collaborative research infrastructure for computational metabolomics. Bioinformatics 31:9 (2015), 1493–1495, 10.1093/bioinformatics/btu813.
Hall, J.E., Dahm, K.H., Seed, J.R., Quantification of tryptophan catabolites from Trypanosoma brucei gambiense in vitro. Comp. Biochem. Physiol. 68B (1981), 521–526.
Hall, J.E., Dahmt, K.H., Seed, J.R., In Vitro tryptophan catabolism by Trypanosoma (Trypanozoon) brucei gambiense, T. (T.) equiperdum, T. (herpetosoma) lewisi and T. (h.) musculi. Comp. 69B (1981), 617–620.
Han, Q., Fang, J., Li, J., Kynurenine aminotransferase and glutamine transaminase K of Escherichia coli: identity with aspartate aminotransferase. Biochem. J. 360:3 (2001), 617–623, 10.1042/0264-6021:3600617.
Han, Qian, Robinson, H., Cai, T., Tagle, D.A., Li, J., Biochemical and structural characterization of mouse mitochondrial aspartate aminotransferase, a newly identified kynurenine aminotransferase-IV. Biosci. Rep. 31:5 (2011), 323–332, 10.1042/BSR20100117.
Hirumi, H., Hirumi, K., Axenic culture of African trypanosome bloodstream forms. Parasitol. Today 10:2 (1994), 80–84, 10.1016/0169-4758(94)90402-2.
Hofer, A., Targeting the nucleotide metabolism of Trypanosoma brucei and other trypanosomatids. FEMS (Fed. Eur. Microbiol. Soc.) Microbiol. Rev., 47(3), 2023, fuad020, 10.1093/femsre/fuad020.
Johnston, K., Kim, D.-H., Kerkhoven, E.J., Burchmore, R., Barrett, M.P., Achcar, F., Mapping the metabolism of five amino acids in bloodstream form Trypanosoma brucei using U-13C-labelled substrates and LC–MS. Biosci. Rep., 39(5), 2019, BSR20181601, 10.1042/BSR20181601.
Karaman, İ., Nørskov, N.P., Yde, C.C., Hedemann, M.S., Bach Knudsen, K.E., Kohler, A., Sparse multi-block PLSR for biomarker discovery when integrating data from LC–MS and NMR metabolomics. Metabolomics 11:2 (2015), 367–379, 10.1007/s11306-014-0698-y.
Kasozi, K.I., MacLeod, E.T., Ntulume, I., Welburn, S.C., An update on african trypanocide pharmaceutics and resistance. Front. Vet. Sci., 9(March), 2022, 10.3389/fvets.2022.828111.
Leclercq, J., Beaufay, C., Catteau, L., Andre, C., Legay, S., METHOD FOR PRODUCING A COMPOSITION COMPRISING A 3-O-p-COUMAROYL ESTER OF TORMENTIC ACID FROM A PLANT CELL CULTURE. APPLICATIONS THEREOF AS ANTIPARASITIC AGENT FOR THE TREATMENT OF TRYPANOSOMIASIS, 2020 https://patents.google.com/patent/WO2020161221A1/en. (Accessed 2 October 2023)
Li, S., Park, Y., Duraisingham, S., Strobel, F.H., Khan, N., Soltow, Q.A., et al. Predicting network activity from high throughput metabolomics. PLoS Comput. Biol., 9(7), 2013, e1003123, 10.1371/journal.pcbi.1003123.
Liao, X., Hu, F., Chen, Z., A HPLC-MS method for profiling triterpenoid acids and triterpenoid esters in Osmanthus fragrans fruits. Analyst 144:23 (2019), 6981–6988, 10.1039/C9AN01542F.
Marchese, L., Nascimento, J.D.F., Damasceno, F.S., Bringaud, F., Michels, P.A.M., Silber, A.M., The uptake and metabolism of amino acids, and their unique role in the biology of pathogenic trypanosomatids. Pathogens 7:2 (2018), 1–45, 10.3390/pathogens7020036.
Marciano, D., Llorente, C., Maugeri, D.A., de la Fuente, C., Opperdoes, F., Cazzulo, J.J., Nowicki, C., Biochemical characterization of stage-specific isoforms of aspartate aminotransferases from Trypanosoma cruzi and Trypanosoma brucei. Mol. Biochem. Parasitol. 161:1 (2008), 12–20, 10.1016/j.molbiopara.2008.05.005.
Marciano, D., Maugeri, D.A., Cazzulo, J.J., Nowicki, C., Functional characterization of stage-specific aminotransferases from trypanosomatids. Mol. Biochem. Parasitol. 166:2 (2009), 172–182, 10.1016/j.molbiopara.2009.04.001.
McGettrick, A.F., Corcoran, S.E., Barry, P.J.G., McFarland, J., Crès, C., Curtis, A.M., et al. Trypanosoma brucei metabolite indolepyruvate decreases HIF-1α and glycolysis in macrophages as a mechanism of innate immune evasion. Proc. Natl. Acad. Sci. U. S. A 113:48 (2016), E7778–E7787, 10.1073/pnas.1608221113.
Michels, P.A.M., Villafraz, O., Pineda, E., Alencar, M.B., Cáceres, A.J., Silber, A.M., Bringaud, F., Carbohydrate metabolism in trypanosomatids: new insights revealing novel complexity, diversity and species-unique features. Exp. Parasitol., 224, 2021, 108102, 10.1016/j.exppara.2021.108102.
Opperdoes, F.R., Butenko, A., Flegontov, P., Yurchenko, V., Lukeš, J., Comparative metabolism of free-living Bodo saltans and parasitic trypanosomatids. J. Eukaryot. Microbiol. 63:5 (2016), 657–678, 10.1111/jeu.12315.
Pink, R., Hudson, A., Mouriès, M.-A., Bendig, M., Opportunities and challenges in antiparasitic drug discovery. Nat. Rev. Drug Discov. 4:9 (2005), 727–740, 10.1038/nrd1824.
Sumner, L.W., Amberg, A., Barrett, D., Beale, M.H., Beger, R., Daykin, C.A., et al. Proposed minimum reporting standards for chemical analysis: chemical analysis working group (CAWG) metabolomics standards initiative (MSI). Metabolomics 3:3 (2007), 211–221, 10.1007/s11306-007-0082-2.
Ternes, C.M., Schönknecht, G., Gene transfers shaped the evolution of de novo NAD+ biosynthesis in eukaryotes. Genome Biol. Evol. 6:9 (2014), 2335–2349, 10.1093/gbe/evu185.
Villani, S., Fallarini, S., Rezzi, S.J., Di Martino, R.M.C., Aprile, S., Del Grosso, E., Selective inhibition of indoleamine and tryptophan 2,3-dioxygenases: comparative study on kynurenine pathway in cell lines via LC-MS/MS-based targeted metabolomics. J. Pharmaceut. Biomed. Anal., 237(June 2023), 2024, 115750, 10.1016/j.jpba.2023.115750.
WHO | Human African trypanosomiasis. (n.d.). WHO. World Health Organization. http://www.who.int/trypanosomiasis_african/en/. Accessed 30 June 2020.
Wiedemar, N., Hauser, D.A., Mäser, P., 100 Years of suramin. Antimicrob. Agents Chemother., 64(3), 2020, 10.1128/AAC.01168-19.
