Aziridine-2,3-Dicarboxylate-Based Cysteine Cathepsin Inhibitors Induce Cell Death in Leishmania major Associated with Accumulation of Debris in Autophagy-Related Lysosome-Like Vacuoles

Schurigt U, Schad C, Glowa C, Baum U, Thomale K, Schnitzer JK, Schultheis M, Schaschke N, Schirmeister T, Moll H (2010)
ANTIMICROBIAL AGENTS AND CHEMOTHERAPY 54(12): 5028-5041.

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Abstract
The papain-like cysteine cathepsins expressed by Leishmania play a key role in the life cycle of these parasites, turning them into attractive targets for the development of new drugs. We previously demonstrated that two compounds of a series of peptidomimetic aziridine-2,3-dicarboxylate [Azi(OBn)(2)]-based inhibitors, Boc-(S)-Leu-(R)-Pro-(S, S)-Azi(OBn)(2) (compound 13b) and Boc-(R)-Leu-(S)-Pro-(S, S)-Azi(OBn)(2) (compound 13e), reduced the growth and viability of Leishmania major and the infection rate of macrophages while not showing cytotoxicity against host cells. In the present study, we characterized the mode of action of inhibitors 13b and 13e in L. major. Both compounds targeted leishmanial cathepsin B-like cysteine cathepsin cysteine proteinase C, as shown by fluorescence proteinase activity assays and active-site labeling with biotin-tagged inhibitors. Furthermore, compounds 13b and 13e were potent inducers of cell death in promastigotes, characterized by cell shrinkage, reduction of mitochondrial transmembrane potential, and increased DNA fragmentation. Transmission electron microscopic studies revealed the enrichment of undigested debris in lysosome-like organelles participating in micro-and macroautophagy-like processes. The release of digestive enzymes into the cytoplasm after rupture of membranes of lysosome-like vacuoles resulted in the significant digestion of intracellular compartments. However, the plasma membrane integrity of compound-treated promastigotes was maintained for several hours. Taken together, our results suggest that the induction of cell death in Leishmania by cysteine cathepsin inhibitors 13b and 13e is different from mammalian apoptosis and is caused by incomplete digestion in autophagy-related lysosome-like vacuoles.
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Schurigt U, Schad C, Glowa C, et al. Aziridine-2,3-Dicarboxylate-Based Cysteine Cathepsin Inhibitors Induce Cell Death in Leishmania major Associated with Accumulation of Debris in Autophagy-Related Lysosome-Like Vacuoles. ANTIMICROBIAL AGENTS AND CHEMOTHERAPY. 2010;54(12):5028-5041.
Schurigt, U., Schad, C., Glowa, C., Baum, U., Thomale, K., Schnitzer, J. K., Schultheis, M., et al. (2010). Aziridine-2,3-Dicarboxylate-Based Cysteine Cathepsin Inhibitors Induce Cell Death in Leishmania major Associated with Accumulation of Debris in Autophagy-Related Lysosome-Like Vacuoles. ANTIMICROBIAL AGENTS AND CHEMOTHERAPY, 54(12), 5028-5041.
Schurigt, U., Schad, C., Glowa, C., Baum, U., Thomale, K., Schnitzer, J. K., Schultheis, M., Schaschke, N., Schirmeister, T., and Moll, H. (2010). Aziridine-2,3-Dicarboxylate-Based Cysteine Cathepsin Inhibitors Induce Cell Death in Leishmania major Associated with Accumulation of Debris in Autophagy-Related Lysosome-Like Vacuoles. ANTIMICROBIAL AGENTS AND CHEMOTHERAPY 54, 5028-5041.
Schurigt, U., et al., 2010. Aziridine-2,3-Dicarboxylate-Based Cysteine Cathepsin Inhibitors Induce Cell Death in Leishmania major Associated with Accumulation of Debris in Autophagy-Related Lysosome-Like Vacuoles. ANTIMICROBIAL AGENTS AND CHEMOTHERAPY, 54(12), p 5028-5041.
U. Schurigt, et al., “Aziridine-2,3-Dicarboxylate-Based Cysteine Cathepsin Inhibitors Induce Cell Death in Leishmania major Associated with Accumulation of Debris in Autophagy-Related Lysosome-Like Vacuoles”, ANTIMICROBIAL AGENTS AND CHEMOTHERAPY, vol. 54, 2010, pp. 5028-5041.
Schurigt, U., Schad, C., Glowa, C., Baum, U., Thomale, K., Schnitzer, J.K., Schultheis, M., Schaschke, N., Schirmeister, T., Moll, H.: Aziridine-2,3-Dicarboxylate-Based Cysteine Cathepsin Inhibitors Induce Cell Death in Leishmania major Associated with Accumulation of Debris in Autophagy-Related Lysosome-Like Vacuoles. ANTIMICROBIAL AGENTS AND CHEMOTHERAPY. 54, 5028-5041 (2010).
Schurigt, Uta, Schad, Caroline, Glowa, Christin, Baum, Ulrike, Thomale, Katja, Schnitzer, Johannes K., Schultheis, Martina, Schaschke, Norbert, Schirmeister, Tanja, and Moll, Heidrun. “Aziridine-2,3-Dicarboxylate-Based Cysteine Cathepsin Inhibitors Induce Cell Death in Leishmania major Associated with Accumulation of Debris in Autophagy-Related Lysosome-Like Vacuoles”. ANTIMICROBIAL AGENTS AND CHEMOTHERAPY 54.12 (2010): 5028-5041.
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11 Citations in Europe PMC

Data provided by Europe PubMed Central.

Development of a New Antileishmanial Aziridine-2,3-Dicarboxylate-Based Inhibitor with High Selectivity for Parasite Cysteine Proteases.
Schad C, Baum U, Frank B, Dietzel U, Mattern F, Gomes C, Ponte-Sucre A, Moll H, Schurigt U, Schirmeister T., Antimicrob. Agents Chemother. 60(2), 2016
PMID: 26596939
Cinnamic Acid Bornyl Ester Derivatives from Valeriana wallichii Exhibit Antileishmanial In Vivo Activity in Leishmania major-Infected BALB/c Mice.
Masic A, Valencia Hernandez AM, Hazra S, Glaser J, Holzgrabe U, Hazra B, Schurigt U., PLoS ONE 10(11), 2015
PMID: 26554591
Autophagic digestion of Leishmania major by host macrophages is associated with differential expression of BNIP3, CTSE, and the miRNAs miR-101c, miR-129, and miR-210.
Frank B, Marcu A, de Oliveira Almeida Petersen AL, Weber H, Stigloher C, Mottram JC, Scholz CJ, Schurigt U., Parasit Vectors 8(), 2015
PMID: 26226952
Cathepsin B in antigen-presenting cells controls mediators of the Th1 immune response during Leishmania major infection.
Gonzalez-Leal IJ, Roger B, Schwarz A, Schirmeister T, Reinheckel T, Lutz MB, Moll H., PLoS Negl Trop Dis 8(9), 2014
PMID: 25255101
Periodate-oxidized ATP modulates macrophage functions during infection with Leishmania amazonensis.
Figliuolo VR, Chaves SP, Santoro GF, Coutinho CM, Meyer-Fernandes JR, Rossi-Bergmann B, Coutinho-Silva R., Cytometry A 85(7), 2014
PMID: 24804957
A novel Leishmania major amastigote assay in 96-well format for rapid drug screening and its use for discovery and evaluation of a new class of leishmanicidal quinolinium salts.
Bringmann G, Thomale K, Bischof S, Schneider C, Schultheis M, Schwarz T, Moll H, Schurigt U., Antimicrob. Agents Chemother. 57(7), 2013
PMID: 23587955
Autophagy in trypanosomatids.
Brennand A, Rico E, Michels PA., Cells 1(3), 2012
PMID: 24710480
Use of cysteine-reactive small molecules in drug discovery for trypanosomal disease.
Nicoll-Griffith DA., Expert Opin Drug Discov 7(4), 2012
PMID: 22458506
Discovery of safe and orally effective 4-aminoquinaldine analogues as apoptotic inducers with activity against experimental visceral leishmaniasis.
Palit P, Hazra A, Maity A, Vijayan RS, Manoharan P, Banerjee S, Mondal NB, Ghoshal N, Ali N., Antimicrob. Agents Chemother. 56(1), 2012
PMID: 22024817
Identification and characterization of genes involved in leishmania pathogenesis: the potential for drug target selection.
Duncan R, Gannavaram S, Dey R, Debrabant A, Lakhal-Naouar I, Nakhasi HL., Mol Biol Int 2011(), 2011
PMID: 22091403

45 References

Data provided by Europe PubMed Central.

Cysteine protease inhibitors containing small rings.
Schirmeister T, Klockow A., Mini Rev Med Chem 3(6), 2003
PMID: 12871161
Generation, culture and flow-cytometric characterization of primary mouse macrophages.
Schleicher U, Bogdan C., Methods Mol. Biol. 531(), 2009
PMID: 19347320

AUTHOR UNKNOWN, 2007
Leishmania major: molecular modeling of cysteine proteases and prediction of new nonpeptide inhibitors.
Selzer PM, Chen X, Chan VJ, Cheng M, Kenyon GL, Kuntz ID, Sakanari JA, Cohen FE, McKerrow JH., Exp. Parasitol. 87(3), 1997
PMID: 9371086
Cysteine protease inhibitors as chemotherapy: lessons from a parasite target.
Selzer PM, Pingel S, Hsieh I, Ugele B, Chan VJ, Engel JC, Bogyo M, Russell DG, Sakanari JA, McKerrow JH., Proc. Natl. Acad. Sci. U.S.A. 96(20), 1999
PMID: 10500116
Synthesis and antileishmanial activities of novel 3-substituted quinolines.
Tempone AG, da Silva AC, Brandt CA, Martinez FS, Borborema SE, da Silveira MA, de Andrade HF Jr., Antimicrob. Agents Chemother. 49(3), 2005
PMID: 15728905
Novel epoxysuccinyl peptides. A selective inhibitor of cathepsin B, in vivo.
Towatari T, Nikawa T, Murata M, Yokoo C, Tamai M, Hanada K, Katunuma N., FEBS Lett. 280(2), 1991
PMID: 2013329
Autophagy-physiology and pathophysiology.
Uchiyama Y, Shibata M, Koike M, Yoshimura K, Sasaki M., Histochem. Cell Biol. 129(4), 2008
PMID: 18320203
Leishmania disease development depends on the presence of apoptotic promastigotes in the virulent inoculum.
van Zandbergen G, Bollinger A, Wenzel A, Kamhawi S, Voll R, Klinger M, Muller A, Holscher C, Herrmann M, Sacks D, Solbach W, Laskay T., Proc. Natl. Acad. Sci. U.S.A. 103(37), 2006
PMID: 16945916

AUTHOR UNKNOWN, 2007
Inhibitors of cysteine proteases.
Vicik R, Busemann M, Baumann K, Schirmeister T., Curr Top Med Chem 6(4), 2006
PMID: 16611146
Aziridide-based inhibitors of cathepsin L: synthesis, inhibition activity, and docking studies.
Vicik R, Busemann M, Gelhaus C, Stiefl N, Scheiber J, Schmitz W, Schulz F, Mladenovic M, Engels B, Leippe M, Baumann K, Schirmeister T., ChemMedChem 1(10), 2006
PMID: 16933358
Aziridine-2,3-dicarboxylate inhibitors targeting the major cysteine protease of Trypanosoma brucei as lead trypanocidal agents.
Vicik R, Hoerr V, Glaser M, Schultheis M, Hansell E, McKerrow JH, Holzgrabe U, Caffrey CR, Ponte-Sucre A, Moll H, Stich A, Schirmeister T., Bioorg. Med. Chem. Lett. 16(10), 2006
PMID: 16516467
Cooperation between apoptotic and viable metacyclics enhances the pathogenesis of Leishmaniasis.
Wanderley JL, Pinto da Silva LH, Deolindo P, Soong L, Borges VM, Prates DB, de Souza AP, Barral A, Balanco JM, do Nascimento MT, Saraiva EM, Barcinski MA., PLoS ONE 4(5), 2009
PMID: 19478944
Cysteine peptidases CPA and CPB are vital for autophagy and differentiation in Leishmania mexicana.
Williams RA, Tetley L, Mottram JC, Coombs GH., Mol. Microbiol. 61(3), 2006
PMID: 16803590
Cell death in Leishmania induced by stress and differentiation: programmed cell death or necrosis?
Zangger H, Mottram JC, Fasel N., Cell Death Differ. 9(10), 2002
PMID: 12232801

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