Abstract
Amoebiasis is caused by the protozoan Entamoeba histolytica that affects millions of people throughout the world. The standard treatment is metronidazole, however, this drug causes several side effects, and is also mutagenic and carcinogenic. Therefore, the search for therapeutic alternatives is necessary. Quinoxaline 1,4-di-N-oxides (QdNOs) derivatives have been shown to exhibit activity against different protozoan. In the present study, the effects of esters of quinoxaline-7-carboxylate 1,4-di-N-oxide (7-carboxylate QdNOs) derivatives on E. histolytica proliferation, morphology, ultrastructure, and oxidative stress were evaluated, also their potential as E. histolytica thioredoxin reductase (EhTrxR) inhibitors was analyzed. In vitro tests showed that 12 compounds from n-propyl and isopropyl series, were more active (IC50 = 0.331 to 3.56 μM) than metronidazole (IC50 = 4.5 μM). The compounds with better biological activity have a bulky, trifluoromethyl and isopropyl group at R1-, R2-, and R3-position, respectively. The main alterations found in trophozoites treated with some of these compounds included changes in chromatin, cell granularity, redistribution of vacuoles with cellular debris, and an increase in reactive oxygen species. Interestingly, docking studies suggested that 7-carboxylate QdNOs derivatives could interact with amino acid residues of the NADPH-binding domain and/or the redox-active site of EhTrxR. Enzymatic assays demonstrated that selected 7-carboxylate QdNOs inhibits EhTrxR disulfide reductase activity, and diaphorase activity shows that these compounds could act as electron acceptor substrates for the enzyme. Taken together, these data indicate that among the mechanisms involved in the antiamoebic effect of the 7-carboxylate QdNOs derivatives studied, is the induction of oxidative stress and the inhibition of EhTrxR activity.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Adil M, Iqbal W, Adnan F, Wazir S, Khan I, Khayam MU, Ahmad S, Ahmed J, Khan IN (2018) Association of Metronidazole with cancer: a potential risk factor or inconsistent deductions? Curr Drug Metab 19:902–909. https://doi.org/10.2174/1389200219666180329124130
Andrade RM, Reed SL (2015) New drug target in protozoan parasites: the role of thioredoxin reductase. Front Microbiol 6:975. https://doi.org/10.3389/fmicb.2015.00975
Arias DG, Regner EL, Iglesias AA, Guerrero SA (2012) Entamoeba histolytica thioredoxin reductase: molecular and functional characterization of its atypical properties. Biochim Biophys Acta 1820:1859–1866. https://doi.org/10.1016/j.bbagen.2012.08.020
Azqueta A, Arbillaga L, Pachón G, Cascante M, Creppy EE, de Cerain AL (2007) A quinoxaline 1, 4-di-N-oxide derivative induces DNA oxidative damage not attenuated by vitamin C and E treatment. Chem Biol Interact 168:95–105. https://doi.org/10.1016/j.cbi.2007.02.013
Bolaños V, Díaz-Martínez A, Soto J, Rodríguez MA, López-Camarillo C, Marchat LA, Ramírez-Moreno E (2014) The flavonoid (−)-epicatechin affects cytoskeleton proteins and functions in Entamoeba histolytica. J Proteome 111:74–85. https://doi.org/10.1016/j.jprot.2014.05.017
Bolaños V, Díaz-Martínez A, Soto J, Marchat LA, Sanchez-Monroy V, Ramírez-Moreno E (2015) Kaempferol inhibits Entamoeba histolytica growth by altering cytoskeletal functions. Mol Biochem Parasitol 204:16–25. https://doi.org/10.1016/j.molbiopara.2015.11.004
Botero DA, Restrepo M (1992) Parasitosis humana. Editorial Presencia Ltda, Bogotá, Colombia; 418 pp. ISBN: 978-958-9076-77-4
Capparelli EV, Bricker-Ford R, Rogers MJ, McKerrow JH, Reed SL (2017) Phase I clinical trial results of auranofin, a novel antiparasitic agent. Antimicrob Agents Chemother 61:e01947–e01916. https://doi.org/10.1128/AAC.01947-16
Carta A, Sanna G, Briguglio I, Madeddu S, Vitale G, Piras S, Corona P, Peana AT, Laurini E, Fermeglia M, Pricl S, Serra A, Carta E, Loddo R, Giliberti G (2018) Quinoxaline derivatives as new inhibitors of coxsackie virus B5. Eur J Med Chem 145:559–569. https://doi.org/10.1016/j.ejmech.2017.12.083
Cedillo-Rivera R, Chavez B, Gonzalez-Robles A, Tapia A, Yepez-Mulia L (2002) In vitro effect of nitazoxanide against Entamoeba histolytica, Giardia intestinalis and Trichomonas vaginalis trophozoites. J Eukaryot Microbiol 49:201–208. https://doi.org/10.1111/j.1550-7408.2002.tb00523.x
Chacón-Vargas KF, Nogueda-Torres B, Sánchez-Torres LE, Suarez-Contreras E, Villalobos-Rocha JC, Torres-Martinez Y, Monge A (2017) Trypanocidal activity of quinoxaline 1,4 Di-N-oxide derivatives as trypanothione reductase inhibitors. Molecules 22:220. https://doi.org/10.3390/molecules22020220
Chacón-Vargas KF, Andrade-Ochoa S, Nogueda-Torres B, Juárez-Ramírez DC, Lara-Ramírez EE, Mondragón-Flores R, Monge A, Rivera G, Sánchez-Torres LE (2018) Isopropyl quinoxaline-7-carboxylate 1, 4-di-N-oxide derivatives induce regulated necrosis-like cell death on Leishmania (Leishmania) mexicana. Parasitol Res 117:45–58. https://doi.org/10.1007/s00436-017-5635-3
Cheng G, Sa W, Cao C, Guo L, Hao H, Liu Z, Wang X, Yuan Z (2016) Quinoxaline 1,4-di-N-oxides: biological activities and mechanisms of actions. Front Pharmacol 7:64. https://doi.org/10.3389/fphar.2016.00064
Das U, Pati HN, Panda AK, De Clercq E, Balzarini J, Molnár J, Baráth Z, Ocsovszki I, Kawase M, Zhou L, Sakagami H, Dimmock JR (2009) 2-(3-Aryl-2-propenoyl)-3-methylquinoxaline-1, 4-dioxides: a novel cluster of tumor-specific cytotoxins which reverse multidrug resistance. Bioorg Med Chem 17(11):3909–3915. https://doi.org/10.1016/j.bmc.2009.04.021
De Chaumont F, Dallongeville S, Chenouard N, Hervé N, Pop S, Provoost T, Meas-Yedid V, Pankajakshan P, Lecomte T, Le Montagner Y, Lagache T, Dufour A, Olivo-Marin JC (2012) Icy: an open bioimage informatics platform for extended reproducible research. Nat Methods 9:690–696. https://doi.org/10.1038/nmeth.2075
Debnath A, Parsonage D, Andrade RM, He C, Cobo ER, Hirata K, Chen S, García-Rivera G, Orozco E, Martínez MB, Gunatilleke SS, Barrios AM, Arkin MR, Poole LB, McKerrow JH, Reed SL (2012) A high-throughput drug screen for Entamoeba histolytica identifies a new lead and target. Nat Med 18:956–960. https://doi.org/10.1038/nm.2758
Diamond LS, Harlow DR, Cunnick CC (1978) A new medium for the axenic cultivation of Entamoeba histolytica and other Entamoeba. Trans R Soc Trop Med Hyg 72:431–432. https://doi.org/10.1016/0035-9203(78)90144-x
Duque-Montaño BE, Gómez-Caro LC, Sanchez-Sanchez M, Monge A, Hernández-Baltazar E, Rivera G, Torres-Angeles O (2013) Synthesis and in vitro evaluation of new ethyl and methyl quinoxaline-7-carboxylate 1, 4-di-N-oxide against Entamoeba histolytica. Bioorg Med Chem 21:4550–4558. https://doi.org/10.1016/j.bmc.2013.05.036
Espinosa A, Socha AM, Ryke E, Rowley DC (2012) Antiamebic properties of the actinomycete metabolites echinomycin A and tirandamycin A. Parasitol Res 111:2473–2477. https://doi.org/10.1007/s00436-012-3019-2
Estevez Y, Quiliano M, Burguete A, Cabanillas B, Zimic M, Málaga E, Verástegui M, Pérez-Silanes S, Aldana I, Monge A, Castillo D, Deharo E (2011) Trypanocidal properties, structure-activity relationship and computational studies of quinoxaline 1,4-di-N-oxide derivatives. Exp Parasitol 127:745–751. https://doi.org/10.1016/j.exppara.2011.01.009
Goes GR, Rocha PS, Diniz AR, Aguiar PH, Machado CR, Vieira LQ (2016) Trypanosoma cruzi needs a signal provided by reactive oxygen species to infect macrophages. PLoS Negl Trop Dis 10:e0004555. https://doi.org/10.1371/journal.pntd.0004555
Gómez-Caro LC, Sánchez-Sánchez M, Bocanegra-García V, Rivera G, Monge A (2011) Synthesis of quinoxaline 1,4-di-N-oxide derivatives on solid support using room temperature and microwave-assisted solvent-free procedures. Quim Nova 34:1147–1151. https://doi.org/10.1590/S0100-40422011000700008
Hanna RM, Dahniya MH, Badr SS, El-Betagy A (2000) Percutaneous catheter drainage in drug-resistant amoebic liver abscess. Tropical Med Int Health 5:578–581. https://doi.org/10.1046/j.1365-3156.2000.00586.x
Haraldsen JD, Liu G, Botting CH, Walton J, Storm J, Phalen T, Kwok L, Soldati-Favre D, Heintz N, Müller S, Westwood N, Ward G (2009) Identification of conoidin A as a covalent inhibitor of peroxiredoxin II. Org Biomol Chem 7:3040–3048. https://doi.org/10.1039/b901735f
Ibrahim MK, Taghour MS, Metwaly AM, Belal A, Mehany ABM, Elhendawy MA, Radwan MM, Yassin AM, El-Deeb NM, Hafez EE, ElSohly MA, Eissa IH (2018) Design, synthesis, molecular modeling and anti-proliferative evaluation of novel quinoxaline derivatives as potential DNA intercalators and topoisomerase II inhibitors. Eur J Med Chem 155:117–134. https://doi.org/10.1016/j.ejmech.2018.06.004
Ishikawa H, Sugiyama T, Yokoyama A (2013) Synthesis of 2,3-Bis(halomethyl)quinoxaline derivatives and evaluation of their antibacterial and antifungal activities. Chem Pharm Bull 61:438–444. https://doi.org/10.1248/cpb.c12-01061
Jaroschik F (2018) Picking one out of three: selective single C−F activation in trifluoromethyl groups. Chem Eur J 24:14572–14582. https://doi.org/10.1002/chem.201801702
Jeelani G, Nozaki T (2016) Entamoeba thiol-based redox metabolism: a potential target for drug development. Mol Biochem Parasitol 206:39–45. https://doi.org/10.1016/j.molbiopara.2016.01.004
Kaplum V, Cogo J, Sangi DP, Ueda-Nakamura T, Corrêa AG, Nakamura CV (2016) In vitro and in vivo activities of 2,3-diarylsubstituted quinoxaline derivatives against Leishmania amazonensis. Antimicrob Agents Chemother 60:3433–3444. https://doi.org/10.1128/AAC.02582-15
Kazy Z, Puhó E, Czeizel AE (2005) Teratogenic potential of vaginal metronidazole treatment during pregnancy. Eur J Obstet Gynecol Reprod Biol 123:174–178. https://doi.org/10.1016/j.ejogrb.2005.03.016
Kelso AA, Goodson SD, Chavan S, Say AF, Turchick A, Sharma D, Ledford LL, Ratterman E, Leskoske K, King AV, Attaway CC, Bandera Y, Foulger SH, Mazin AV, Temesvari LA, Sehorn MG (2016) Characterization of the recombination activities of the Entamoeba histolytica Rad51 recombinase. Mol Biochem Parasitol 210:71–84. https://doi.org/10.1016/j.molbiopara.2016.09.001
Lee SH, Kim N, Kim SJ, Song J, Gong YD, Kim SY (2013) Anti-cancer effect of a quinoxaline derivative GK13 as a transglutaminase 2 inhibitor. J Cancer Res Clin Oncol 139:1279–1294. https://doi.org/10.1007/s00432-013-1433-1
Leitsch D, Kolarich D, Wilson IB, Altmann F, Duchêne M (2007) Nitroimidazole action in Entamoeba histolytica: a central role for thioredoxin reductase. PLoS Biol 5:e211. https://doi.org/10.1371/journal.pbio.0050211
Moreno-Viguri E, Pérez-Silanes S (2013) Quinoxaline 1,4-di-N-oxide derivatives: interest in the treatment of chagas disease. Rev Virtual Quim 5:1101–1119. https://doi.org/10.5935/1984-6835.20130080
Nagaraja S, Ankri S (2018) Utilization of different omic approaches to unravel stress response mechanisms in the parasite Entamoeba histolytica. Front Cell Infect Microbiol 8:19. https://doi.org/10.3389/fcimb.2018.00019
Nandi N, Sen A, Banerjee R, Kumar S, Kumar V, Ghosh AN, Das P (2010) Hydrogen peroxide induces apoptosis-like death in Entamoeba histolytica trophozoites. Microbiology 156:1926–1941. https://doi.org/10.1099/mic.0.034066-0
Pais-Morales J, Betanzos A, García-Rivera G, Chávez-Munguía B, Shibayama M, Orozco E (2016) Resveratrol induces apoptosis-like death and prevents in vitro and in vivo virulence of Entamoeba histolytica. PLoS One 11:e0146287. https://doi.org/10.1371/journal.pone.0146287
Parsonage D, Sheng F, Hirata K, Debnath A, McKerrow JH, Reed SL, Abagyan R, Poole LB, Podust LM (2016) X-ray structures of thioredoxin and thioredoxin reductase from Entamoeba histolytica and prevailing hypothesis of the mechanism of Auranofin action. J Struct Biol 194:180–190. https://doi.org/10.1016/j.jsb.2016.02.015
Patel SB, Patel BD, Pannecouque C, Bhatt HG (2016) Design, synthesis and anti-HIV activity of novel quinoxaline derivatives. Eur J Med Chem 117:230–240. https://doi.org/10.1016/j.ejmech.2016.04.019
Perdomo D, Aït-Ammar N, Syan S, Sachse M, Jhingan GD, Guillén N (2015) Cellular and proteomics analysis of the endomembrane system from the unicellular Entamoeba histolytica. J Proteome 112:125–140. https://doi.org/10.1016/j.jprot.2014.07.034
Pineda E, Perdomo D (2017) Entamoeba histolytica under oxidative stress: what countermeasure mechanisms are in place? Cells 6:44. https://doi.org/10.3390/cells6040044
Pizzino G, Irrera N, Cucinotta M, Pallio G, Mannino F, Arcoraci V, Squadrito F, Altavilla D, Bitto A (2017) Oxidative stress: harms and benefits for human health. Oxidative Med Cell Longev:8416763. https://doi.org/10.1155/2017/8416763
Proshin AN, Orlova MA, Trofimova TP (2017) Biological activity of some sulfur-and selenium-containing spiro compounds. Russ Chem Bull 66:1931–1933. https://doi.org/10.1007/s11172-017-1968-4
Rodrigues JH, Ueda-Nakamura T, Corrêa AG, Sangi DP, Nakamura CV (2014) A quinoxaline derivative as a potent chemotherapeutic agent, alone or in combination with benznidazole, against Trypanosoma cruzi. PLoS One 9:e85706. https://doi.org/10.1371/journal.pone.0085706 eCollection 2014
Schlosser S, Leitsch D, Duchêne M (2013) Entamoeba histolytica: identification of thioredoxin-targeted proteins and analysis of serine acetyltransferase-1 as a prototype example. Biochem J 451:277–288. https://doi.org/10.1042/BJ20121798
Shahi P, Chadee K (2017) Human amebiasis: insight into the biology and immunopathogenesis. Springer, Cham. Neglected tropical diseases - South Asia. pp 65–82
Shahi P, Trebicz-Geffen M, Nagaraja S, Alterzon-Baumel S, Hertz R, Methling K, Lalk M, Ankri S (2016) Proteomic identification of oxidized proteins in Entamoeba histolytica by resin-assisted capture: insights into the role of Arginase in resistance to oxidative stress. PLoS Negl Trop Dis 10:e0004340. https://doi.org/10.1371/journal.pntd.0004340
Sharma P, Jha AB, Dubey RS, Pessarakli M (2012) Reactive oxygen species, oxidative damage, and antioxidative defense mechanism in plants under stressful conditions. Aust J Bot 2012:1–26. https://doi.org/10.1155/2012/217037
Villalba JD, Gómez C, Medel O, Sánchez V, Carrero JC, Shibayama M, Ishiwara DG (2007) Programmed cell death in Entamoeba histolytica induced by the aminoglycoside G418. Microbiology 153:3852–3863. https://doi.org/10.1099/mic.0.2007/008599-0
Villalobos-Rocha JC, Sánchez-Torres L, Nogueda-Torres B, Segura-Cabrera A, García-Pérez CA, Bocanegra-García V, Palos I, Monge A, Rivera G (2014) Anti-Trypanosoma cruzi and anti-leishmanial activity by quinoxaline-7-carboxylate 1,4-di-N-oxide derivatives. Parasitol Res 113:2027–2035. https://doi.org/10.1007/s00436-014-3850-8
Wang X, Martínez MA, Cheng G, Liu Z, Huang L, Dai M, Chen D, Martínez-Larrañaga MR, Anadón A, Yuan Z (2016) The critical role of oxidative stress in the toxicity and metabolism of quinoxaline 1,4-di-N-oxides in vitro and in vivo. Drug Metab Rev 48:159–182. https://doi.org/10.1080/03602532.2016.1189560
You L, Cho EJ, Leavitt J, Ma LC, Montelione GT, Anslyn EV, Krug RM, Ellington A, Robertus JD (2011) Synthesis and evaluation of quinoxaline derivatives as potential influenza NS1A protein inhibitors. Bioorg Med Chem Lett 21:3007–3011. https://doi.org/10.1016/j.bmcl.2011.03.042
Zavala-Ocampo LM, Aguirre-Hernández E, Pérez-Hernández N, Rivera G, Marchat LA, Ramírez-Moreno E (2017) Antiamoebic activity of Petiveria alliacea leaves and their main component, isoarborinol. J Microbiol Biotechnol 27:1401–1408. https://doi.org/10.4014/jmb.1705.05003
Zhao Y, Cheng G, Hao H, Pan Y, Liu Z, Dai M, Yuan Z (2016) In vitro antimicrobial activities of animal-used quinoxaline 1,4-di-N-oxides against mycobacteria, mycoplasma and fungi. BMC Vet Res 12:186. https://doi.org/10.1186/s12917-016-0812-7
Acknowledgments
We thank Oliver Ornelas-Lopez and Maria del Rosario Espinoza Mellado, from the Microscopy Unit of ENCB, Mexico for their technical help in TEM assays.
Funding
This work was supported by the Instituto Politécnico Nacional, Mexico (SIP-20180520 and SIP-20190295); Consejo Nacional de Ciencia y Tecnología (CONACYT), Mexico; and ANPCyT (PICT2016-1778), Argentina.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
On behalf of all authors, the corresponding author states that there is no conflict of interest.
Additional information
Section Editor: Yaoyu Feng
Publisher’s note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Appendix
Appendix
Predicted model showing the interaction of E. histolytica thioredoxin reductase (EhTrxR) with a Auranofin and b Metronidazole. 3D representation (left) and 2D Ligplot (right) showing protein-ligand interactions during docking. Hydrogen bonds are highlighted in green with the respective distances between the atoms involved, whereas hydrophobic contacts are represented by an arc with spokes radiating towards the ligand atom
EhTrxR disulfide reductase activity inhibition profiles. The assays were performed at 30 °C and pH 7.0 in the presence of 5 mM DTNB, 200 μM NADPH, 0.1 μM EhTrxR, and different concentrations of T-001, T-067, or T-116. The activities were normalized to measurements performed in the absence of the compounds
Rights and permissions
About this article
Cite this article
Soto-Sánchez, J., Caro-Gómez, L.A., Paz-González, A.D. et al. Biological activity of esters of quinoxaline-7-carboxylate 1,4-di-N-oxide against E. histolytica and their analysis as potential thioredoxin reductase inhibitors. Parasitol Res 119, 695–711 (2020). https://doi.org/10.1007/s00436-019-06580-8
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00436-019-06580-8