Home Medicine Asymmetric peptidomimetics containing L-tartaric acid core inhibit the aspartyl peptidase activity and growth of Leishmania amazonensis promastigotes
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Asymmetric peptidomimetics containing L-tartaric acid core inhibit the aspartyl peptidase activity and growth of Leishmania amazonensis promastigotes

  • André L.S. Santos EMAIL logo , Filipe P. Matteoli , Leandro S. Sangenito , Marta H. Branquinha , Bruno A. Cotrim and Gabriel O. Resende
Published/Copyright: January 17, 2018
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Acta Parasitologica
From the journal Acta Parasitologica Volume 63 Issue 1

Abstract

Aspartyl-type peptidases are promising chemotherapeutic targets in protozoan parasites. In the present work, we identified an aspartyl peptidase activity from the soluble extract of Leishmania amazonensis promastigotes, which cleaved the fluorogenic peptide 7-methoxycoumarin-4-acetyl-Gly-Lys-Pro-Ile-Leu-Phe-Phe-Arg-Leu-Lys(DNP)-D-Arg-amide (cathepsin D substrate) under acidic pH conditions at 37°C, showing a KM of 0.58 μM and Vmax of 129.87 fluorescence arbitrary units/s mg protein. The leishmanial aspartyl peptidase activity was blocked by pepstatin A (IC50 = 6.8 μM) and diazo-acetyl-norleucinemetilester (IC50 = 10.2 μM), two classical aspartyl peptidase inhibitors. Subsequently, the effects of 6 asymmetric peptidomimetics, containing L-tartaric acid core, were tested on both aspartyl peptidase and growth of L. amazonensis promastigotes. The peptidomimetics named 88, 154 and 158 promoted a reduction of 50% on the leishmanial aspartyl peptidase activity at concentrations ranging from 40 to 85 μM, whereas the peptidomimetic 157 was by far the most effective, presenting IC50 of 0.04 μM. Furthermore, the peptidomimetics 157 and 154 reduced the parasite proliferation in a dose-dependent manner, displaying IC50 values of 33.7 and 44.5 μM, respectively. Collectively, the peptidomimetic 157 was the most efficient compound able to arrest both aspartyl peptidase activity and leishmanial proliferation, which raises excellent perspectives regarding its use against this human pathogenic protozoan.

Keywords: Leishmania amazonensis; peptidomimetics; L-tartaric acid; aspartyl peptidases; aspartyl peptidase inhibitors; anti-Leishmania action

  1. Funding: This work was supported by grants from Fundação Carlos Chagas Filho de Amparo á Pesquisa do Estado do Rio de Janeiro (FAPERJ), Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES).

  2. Conflict of interest: The authors have no conflicts of interest to declare.

Acknowledgment

The authors would like to thank Denise Rocha de Souza, who is supported by FAPERJ scholarship, for her technical assistance.

References

Abrahim-Vieira B., da Costa E.C.B., de Azevedo P.H.R., Portela A.C., Dias L.R.S., Pinheiro S., et al. 2014. Novel isomannide-based peptide mimetics containing a tartaric acid backbone as serine protease inhibitors. Medicinal Chemistry Research, 23, 5305–5320. 10.1007/s00044-014-1058-1Search in Google Scholar

Alfonso Y., Monzote L. 2011. HIV protease inhibitors: effect on the opportunistic protozoan parasites. The Open Medicinal Chemistry Journal, 5, 40–50. 10.2174/1874104501105010040Search in Google Scholar PubMed PubMed Central

Alves C.R., Corte-Real S., Bourguignon S.C., Chaves C.S., Saraiva E.M. 2005. Leishmania amazonensis: early proteinase activities during promastigote-amastigote differentiation in vitro, Experimental Parasitology, 109, 38–48. 10.1016/j.exppara.2004.10.005Search in Google Scholar PubMed

Barros J.C., da Silva J.F.M., Calazans A., Tanuri A., Brindeiro R.M., Williamson J.S., et al. 2006. Synthesis of pseudopeptides derived from (R, R)-tartaric acid as potential inhibitors of HIV-protease. Letters in Organic Chemistry, 3, 882–88610.2174/157017806779468040Search in Google Scholar

Bastos I.M., Motta F.N., Grellier P., Santana J.M. 2013. Parasite prolyl oligopeptidases and the challenge of designing chemother-apeuticals for Chagas disease, leishmaniasis and African trypanosomiasis. Current Medicinal Chemistry, 20, 3103–3115. 10.2174/0929867311320250006Search in Google Scholar PubMed PubMed Central

Bates P.A., Robertson C.D., Coombs G.H. 1994. Expression of cysteine proteinases by metacyclic promastigotes of Leishmania mexicana. Journal of Eukaryotic Microbiology, 41, 199–203. 10.1111/j.1550-7408Search in Google Scholar

Branquinha M.H., Sangenito L.S., Sodré C.L., Kneipp L.F., d’ Avila-Levy C.M., Santos A.L.S. 2017. The widespread anti-protozoal action of HIV aspartic peptidase inhibitors: focus on Plasmodium spp., Leishmania spp. and Trypanosoma cruzi. Current Topics in Medicinal Chemistry, 17, 1303–1317. 10.2174/1568026616666161025161153Search in Google Scholar PubMed

Caffrey C.R., Lima A.P., Steverding D. 2011. Cysteine peptidases of kinetoplastid parasites, Advances in Experimental Medicine and Biology, 712, 84–99. 10.1007/978-1-4419-8414-2_6Search in Google Scholar PubMed

Coates L., Erskine P.T., Mall S., Gill R., Wood S.P., Myles D.A., et al. 2006. X-Ray, neutron and NMR studies of the catalytic mechanism of aspartic proteinases. European Biophysics Journal, 35, 559–566. 10.1007/s00249-006-0065-7Search in Google Scholar PubMed

Dali B., Keita M., Megnassan E., Frecer V., Miertus S. 2012. Insight into selectivity of peptidomimetic inhibitors with modified statine core for plasmepsin II of Plasmodium falciparum over human cathepsin D. Chemical Biology and Drug Design, 79, 411–430. 10.1111/j.1747-0285.2011.01276.xSearch in Google Scholar PubMed

Dash C., Kulkarni A., Dunn B., Rao M. 2003. Aspartic peptidase inhibitors: implications in drug development. Critical Reviews in Biochemistry and Molecular Biology, 38, 89–119. 10.1080/713609213Search in Google Scholar PubMed

Davies D.R. 1990. The structure and function of the aspartic proteinases. Annual Review of Biophysics and Biophysical Chemistry, 19, 189–215. 10.1146/annurev.bb.19.060190.001201Search in Google Scholar PubMed

Dominguez D.I., Hartmann D., De Strooper B. 2004. BACE1 and presenilin: two unusual aspartic proteases involved in Alzheimer’s disease. Neurodegenerative Disease, 1, 168–174. 10.1159/000080982Search in Google Scholar PubMed

Eder J., Hommel U., Cumin F., Martoglio B., Gerhartz B. 2007. Aspartic proteases in drug discovery. Current Pharmaceutical Design. 13, 271–285. 10.2174/138161207779313560Search in Google Scholar PubMed

Gazdik M., Jarman K.E., O’Neill M.T., Hodder A.N., Lowes K.N., Jousset Sabroux H., et al. 2016. Exploration of the P3 region of PEXEL peptidomimetics leads to a potent inhibitor of the Plasmodium protease, plasmepsin V. Bioorganic and Medicinal Chemistry, 24, 1993–2010. 10.1016/j.bmc.2016.03.027Search in Google Scholar PubMed

Gonzalez A.A., Prieto M.C. 2015. Renin and the (pro)renin receptor in the renal collecting duct: role in the pathogenesis of hypertension. Clinical and Experimental Pharmacology and Physiology, 42, 14–21. 10.1111/1440-1681.12319Search in Google Scholar PubMed PubMed Central

Hamada Y., Kiso Y. 2016. New directions for protease inhibitors directed drug discovery. Biopolymers, 106, 563–579. 10.1002/bip.22780Search in Google Scholar PubMed PubMed Central

Harris J.L., Backes B.J., Leonetti F., Mahrus S., Ellman J.A., Craik C.S. 2000. Rapid and general profiling of protease specificity by using combinatorial fluorogenic substrate libraries. Proceedings of the National Academy of Sciences of the United States of America, 97, 7754–7759. 10.1073/pnas.140132697Search in Google Scholar PubMed PubMed Central

Hemández-Chinea C., Maimone L., Campos Y., Mosca W., Romero P.J. 2017. Apparent isocitrate lyase activity in Leishmania amazonensis. Acta Parasitologica, 62, 701–707. 10.1515/ap-2017-0084Search in Google Scholar PubMed

Horimoto Y., Dee D.R., Yada R.Y. 2009. Multifunctional aspartic peptidase prosegments. New Biotechnology, 25, 318–324. 10.1016/j.nbt.2009.03.010Search in Google Scholar PubMed

Koehler J.W., Morales M.E., Shelby B.D., Brindley P.J. 2007. Aspartic protease activities of schistosomes cleave mammalian hemoglobins in a host-specific manner. Memórias do Instituto Oswaldo Cruz, 102, 83–8510.1590/S0074-02762007000100014Search in Google Scholar

Lima A.K.C., Elias C.G.R., Souza J.E.O., Santos A.L.S., Dutra P.M.L. 2009. Dissimilar peptidase production by avirulent and virulent promastigotes of Leishmania braziliensis: inference on the parasite proliferation and interaction with macrophages. Parasitology, 136, 1179–1191. 10.1017/S0031182009990540Search in Google Scholar PubMed

Lowry O.H., Rosebrough N.J., Farr A.L., Randal R.J. 1951. Protein measurement with the folin phenol reagent. Journal of Biological Chemistry, 193, 264–27510.1016/S0021-9258(19)52451-6Search in Google Scholar

Majer F., Pavlícková L., Majer P., Hradilek M., Dolejsí E., Hrusková-Heidingsfeldová O., et al. 2006. Structure-based specificity mapping of secreted aspartic proteases of Candida parapsilosis, Candida albicans, and Candida tropicalis using peptidomimetic inhibitors and homology modeling. Biological Chemistry, 387, 1247–1254. 10.1515/BC.2006.154Search in Google Scholar PubMed

Nielsen P.E. 2004. Pseudo-peptides in drug discovery. John Wiley and Sons, ISBN: 978-3-527, 30633-30636Search in Google Scholar

Olivier M., Atayde V.D., Isnard A., Hassani K., Shio M.T. 2012. Leishmania virulence factors: focus on the metalloprotease GP63. Microbes and Infection, 14, 1377–1389. 10.1016/j.micinf.2012.05.014Search in Google Scholar PubMed

Peçanha E.P., Figueiredo L.J., Brindeiro R.M., Tanuri A., Calazans A.R., Antunes O.A. 2003. Synthesis and anti-HIV activity of new C2 symmetric derivatives designed as HIV-1 protease inhibitors. Farmaco, 58, 149–157. org/10.1016/S0014-827X(02)00016-2Search in Google Scholar PubMed

Perteguer M.J., Gómez-Puertas P., Cañavate C., Dagger F., Gárate T., Valdivieso E. 2012. Ddi1-like protein from Leishmania major is an active aspartyl proteinase. Cell Stress Chaperones, 18, 171–181. 10.1007/s12192-012-0368-9Search in Google Scholar PubMed PubMed Central

Pranjol M.Z., Gutowski N., Hannemann M., Whatmore J. 2015. The potential role of the proteases cathepsin D and cathepsin L in the progression and metastasis of epithelial ovarian cancer. Biomolecules, 5, 3260–3279. 10.3390/biom5043260Search in Google Scholar PubMed PubMed Central

Qidwai T. 2015. Hemoglobin Degrading proteases of Plasmodium falciparum as antimalarial drug targets. Current Drug Targets, 16, 1133–1141. 10.2174/1389450116666150304104123Search in Google Scholar PubMed

Qiu X., Liu Z.P. 2011. Recent developments of peptidomimetic HIV-1 protease inhibitors. Current Medicinal Chemistry, 18, 4513–4537. 10.2174/092986711797287566Search in Google Scholar PubMed

Resende G.O., João F.C., da Silva F.C., Cotrim B.A., Antunes O.A.C., Aguiar L.C.S. 2007. Synthesis of asymmetric peptide mimetic compounds containing tartaric acid core. Potential inhibitors of HIV-1 protease. Letters in Organic Chemistry, 4, 146–150. 10.2174/157017807780737200Search in Google Scholar

Sádlová J., Volf P., Victoir K., Dujardin J., Votypka J. 2006. Virulent and attenuated lines of Leishmania major: DNA karyotypes and differences in metalloproteinase GP63. Folia Parasitologica, 53, 81–90. 10.14411/fp.2006.011Search in Google Scholar PubMed

Sangenito L.S., Gonçalves D.S., Seabra S.H., d’Avila-Levy C.M., Santos A.L.S., Branquinha M.H. 2016. HIV aspartic peptidase inhibitors are effective drugs against the trypomastigote form of the human pathogen Trypanosoma cruzi. International Journal of Antimicrobial Agents, 48, 440–444. 10.1016/j.ijantimicag.2016.06.024Search in Google Scholar PubMed

Santos L.O., Marinho F.A., Altoé E.F., Vitório B.S., Alves C.R., Britto C., et al. 2009. HIV aspartyl peptidase inhibitors interfere with cellular proliferation, ultrastructure and macrophage infection of Leishmania amazonensis. PLoS ONE, 4, e4918. 10.1371/journal.pone.0004918Search in Google Scholar PubMed PubMed Central

Santos L.O., Garcia-Gomes A.S., Catanho M., Sodre C.L., Santos A.L.S., Branquinha M.H., et al. 2013a. Aspartic peptidases of human pathogenic trypanosomatids: perspectives and trends for chemotherapy. Current Medicinal Chemistry, 20, 3116–3333. 10.2174/0929867311320250007Search in Google Scholar PubMed PubMed Central

Santos L.O., Vitório B.S., Branquinha M.H., Pedrozo C., Santos A.L.S., d’Avila-Levy C.M. 2013b. Nelfinavir, an HIV aspartyl peptidase inhibitor, is effective in inhibiting the multiplication and aspartyl peptidase activity of several Leishmania species, including strains obtained from HIV-positive patients. Journal of Antmicrobial Chemotherapy, 68, 348–353. 10.1093/jac/dks410Search in Google Scholar PubMed PubMed Central

Savoia D. 2015. Recent updates and perspectives on leishmaniasis. The Journal of Infection in Developing Countries, 9, 588–596. 10.3855/jidc.6833Search in Google Scholar PubMed

Silva N.C., Nery J.M., Dias A.L. 2014. Aspartic proteinases of Candida spp.: role in pathogenicity and antifungal resistance. Mycoses, 57, 1–11. 10.1111/myc.12095Search in Google Scholar PubMed

Sojka D., Hartmann D., Bartošová-Sojková P., Dvořák J. 2016. Parasite cathepsin D-like peptidases and their relevance as therapeutic targets. Trends in Parasitology, 32, 708–723. 10.1016/j.pt.2016.05.015Search in Google Scholar PubMed

Trudel N., Garg R., Messier N., Sundar S., Ouellette S.M., Tremblay M.J. 2008. Intracellular survival of Leishmania species that cause visceral leishmaniasis is significantly reduced by HIV-1 protease inhibitors. The Journal of Infectious Diseases, 198, 1292–1299. 10.1086/592280Search in Google Scholar PubMed

Valdivieso E., Dagger F., Rascón A. 2007. Leishmania mexicana: identification and characterization of an aspartic proteinase activity. Experimental Parasitology, 116, 77–82E. 10.1016/j.exppara.2006.10.006Search in Google Scholar PubMed

Valdivieso A., Rangel J., Moreno J.M., Saugar C., Cañavate J., Alvar F., et al. 2010. Effects of HIV aspartyl proteinase inhibitors on Leishmania sp., Experimental Parasitology, 126, 557–563. 10.1016/j.exppara.2010.06.002Search in Google Scholar PubMed

Wensing A.M., Van Maarseveen N.M., Nijhuis M. 2010. Fifteen years of HIV protease inhibitors: raising the barrier to resistance. Antiviral Research, 85, 59–74. 10.1016/j.antiviral.2009.10.003Search in Google Scholar PubMed

Received: 2017-7-29
Revised: 2017-10-30
Accepted: 2017-11-10
Published Online: 2018-1-17
Published in Print: 2018-3-26

© 2018 W. Stefański Institute of Parasitology, PAS

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https://doi.org/10.1515/ap-2018-0013
Keywords for this article
Leishmania amazonensis; peptidomimetics; L-tartaric acid; aspartyl peptidases; aspartyl peptidase inhibitors; anti-Leishmania action

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  12. Molecular target analysis of stearoyl-CoA desaturase genes of protozoan parasites
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  22. Metazoan parasite fauna of migrating common garfish, Belone belone (L.), in the Baltic Sea
  24. Measurement of binding strength between prey proteins interacting with Toxoplasma gondii SAG1 and SAG2 using isothermal titration calorimetry (ITC)
  26. Asymmetric peptidomimetics containing L-tartaric acid core inhibit the aspartyl peptidase activity and growth of Leishmania amazonensis promastigotes
  28. Transcriptional analysis of immune-relevant genes in the mucus of Labeo rohita, experimentally infected with Argulus siamensis
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