#PAGE_PARAMS# #ADS_HEAD_SCRIPTS# #MICRODATA#

Identification of Plasmodium dipeptidyl aminopeptidase allosteric inhibitors by high throughput screening


Autoři: Mateo I. Sanchez aff001;  Laura E. de Vries aff002;  Christine Lehmann aff002;  Jeong T. Lee aff001;  Kenny K. Ang aff003;  Christopher Wilson aff003;  Steven Chen aff003;  Michelle R. Arkin aff003;  Matthew Bogyo aff001;  Edgar Deu aff002
Působiště autorů: Departments of Pathology and Microbiology & Immunology, Stanford School of Medicine, Stanford, CA, United States of America aff001;  Chemical Biology Approaches to Malaria Lab, The Francis Crick Institute, London, United Kingdom aff002;  Small Molecule Discovery Center and Department of Pharmaceutical Chemistry, University of California at San Francisco, San Francisco, CA, United States of America aff003
Vyšlo v časopise: PLoS ONE 14(12)
Kategorie: Research Article
doi: https://doi.org/10.1371/journal.pone.0226270

Souhrn

Dipeptidyl aminopeptidases (DPAPs) are cysteine proteases that cleave dipeptides from the N-terminus of protein substrates and have been shown to play important roles in many pathologies including parasitic diseases such as malaria, toxoplasmosis and Chagas’s disease. Inhibitors of the mammalian homologue cathepsin C have been used in clinical trials as potential drugs to treat chronic inflammatory disorders, thus proving that these enzymes are druggable. In Plasmodium species, DPAPs play important functions at different stages of parasite development, thus making them potential antimalarial targets. Most DPAP inhibitors developed to date are peptide-based or peptidomimetic competitive inhibitors. Here, we used a high throughput screening approach to identify novel inhibitor scaffolds that block the activity of Plasmodium falciparum DPAP1. Most of the hits identified in this screen also inhibit Plasmodium falciparum DPAP3, cathepsin C, and to a lesser extent other malarial clan CA proteases, indicating that these might be general DPAP inhibitors. Interestingly, our mechanism of inhibition studies indicate that most hits are allosteric inhibitors, which opens a completely new strategy to inhibit these enzymes, study their biological function, and potentially develop new inhibitors as starting points for drug development.

Klíčová slova:

Antimalarials – Enzyme inhibitors – Malaria – Malarial parasites – Parasitic diseases – Plasmodium – Proteases – Parasite replication


Zdroje

1. World Health Organization. World Malaria Report 2016. 2017;: 1–186.

2. Ashley EA, Dhorda M, Fairhurst RM, Amaratunga C, Lim P, Suon S, et al. Spread of artemisinin resistance in Plasmodium falciparum malaria. N Engl J Med. 2014;371: 411–423. doi: 10.1056/NEJMoa1314981 25075834

3. Burrows JN, Duparc S, Gutteridge WE, van Huijsduijnen RH, Kaszubska W, Macintyre F, et al. New developments in anti-malarial target candidate and product profiles. Malar J. 2017;16: 1–29. doi: 10.1186/s12936-016-1650-6

4. Wells TNC, Hooft van Huijsduijnen R, Van Voorhis WC. Malaria medicines: a glass half full? Nat Rev Drug Discov. 2015;14: 424–442. doi: 10.1038/nrd4573 26000721

5. Turk B. Targeting proteases: successes, failures and future prospects. Nat Rev Drug Discov. 2006;5: 785–799. doi: 10.1038/nrd2092 16955069

6. Drag M, Salvesen GS. Emerging principles in protease-based drug discovery. Nat Rev Drug Discov. 2010;9: 690–701. doi: 10.1038/nrd3053 20811381

7. Deu E. Proteases as antimalarial targets: strategies for genetic, chemical, and therapeutic validation. FEBS J. 2017;284: 2604–2628. doi: 10.1111/febs.14130 28599096

8. Tanaka TQ, Deu E, Molina-Cruz A, Ashburne MJ, Ali O, Suri A, et al. Plasmodium dipeptidyl aminopeptidases as malaria transmission-blocking drug targets. Antimicrob Agents Chemother. 2013;57: 4645–4652. doi: 10.1128/AAC.02495-12 23836185

9. Suárez-Cortés P, Sharma V, Bertuccini L, Costa G, Bannerman N-L, Sannella AR, et al. Comparative proteomics and functional analysis reveal a role of Plasmodium falciparum osmiophilic bodies in malaria parasite transmission. Mol Cell Proteomics. 2016;15: 3243–3255. doi: 10.1074/mcp.M116.060681 27432909

10. Klemba M, Gluzman I, Goldberg DE. A Plasmodium falciparum dipeptidyl aminopeptidase I participates in vacuolar hemoglobin degradation. J Biol Chem. 2004;279: 43000–43007. doi: 10.1074/jbc.M408123200 15304495

11. Deu E, Leyva MJ, Albrow VE, Rice MJ, Ellman JA, Bogyo M. Functional studies of Plasmodium falciparum dipeptidyl aminopeptidase I using small molecule inhibitors and active site probes. Chem Biol. 2010;17: 808–819. doi: 10.1016/j.chembiol.2010.06.007 20797610

12. Lehmann C, Tan MSY, de Vries LE, Russo I, Sanchez MI, Goldberg DE, et al. Plasmodium falciparum dipeptidyl aminopeptidase 3 activity is important for efficient erythrocyte invasion by the malaria parasite. PLoS Pathog. 2018;14: e1007031. doi: 10.1371/journal.ppat.1007031 29768491

13. Wiggans DS, Winitz M, Fruton JS. Action of cathepsin C on dipeptide esters. Yale J Biol Med. 1954;27: 11–19. 13196397

14. Gutmann HR, Fruton JS. On the proteolytic enzymes of animal tissues; an intracellular enzyme related to chymotrypsin. J Biol Chem. 1948;174: 851–858. 18871244

15. Lainé D, Palovich M, McCleland B, Petitjean E, Delhom I, Xie H, et al. Discovery of novel cyanamide-based inhibitors of cathepsin C. ACS Med Chem Lett. 2011;2: 142–147. doi: 10.1021/ml100212k 24900293

16. Guay D, Beaulieu C, Truchon J-F, Jagadeeswar Reddy T, Zamboni R, Bayly CI, et al. Design and synthesis of dipeptidyl nitriles as potent, selective, and reversible inhibitors of cathepsin C. Bioorg Med Chem Lett. 2009;19: 5392–5396. doi: 10.1016/j.bmcl.2009.07.114 19665376

17. Furber M, Tiden A-K, Gardiner P, Mete A, Ford R, Millichip I, et al. Cathepsin C inhibitors: property optimization and identification of a clinical candidate. J Med Chem. 2014;57: 2357–2367. doi: 10.1021/jm401705g 24592859

18. Korkmaz B, Caughey GH, Chapple I, Gauthier F, Hirschfeld J, Jenne DE, et al. Therapeutic targeting of cathepsin C: from pathophysiology to treatment. Pharmacol Ther. 2018;190: 202–236. doi: 10.1016/j.pharmthera.2018.05.011 29842917

19. McGuire MJ, Lipsky PE, Thiele DL. Generation of active myeloid and lymphoid granule serine proteases requires processing by the granule thiol protease dipeptidyl peptidase I. J Biol Chem. 1993;268: 2458–2467. 8428921

20. Kummer JA, Kamp AM, Citarella F, Horrevoets AJ, Hack CE. Expression of human recombinant granzyme A zymogen and its activation by the cysteine proteinase cathepsin C. J Biol Chem. 1996;271: 9281–9286. doi: 10.1074/jbc.271.16.9281 8621589

21. Pham CT, Ley TJ. Dipeptidyl peptidase I is required for the processing and activation of granzymes A and B in vivo. Proc Natl Acad Sci USA. 1999;96: 8627–8632. doi: 10.1073/pnas.96.15.8627 10411926

22. Adkison AM, Raptis SZ, Kelley DG, Pham CTN. Dipeptidyl peptidase I activates neutrophil-derived serine proteases and regulates the development of acute experimental arthritis. J Clin Invest. 2002;109: 363–371. doi: 10.1172/JCI13462 11827996

23. Miller BE, Mayer RJ, Goyal N, Bal J, Dallow N, Boyce M, et al. Epithelial desquamation observed in a phase I study of an oral cathepsin C inhibitor (GSK2793660). Br J Clin Pharmacol. 2017;83: 2813–2820. doi: 10.1111/bcp.13398 28800383

24. Palmér R, Mäenpää J, Jauhiainen A, Larsson B, Mo J, Russell M, et al. Dipeptidyl peptidase 1 Inhibitor AZD7986 induces a sustained, exposure-dependent reduction in neutrophil elastase activity in healthy subjects. Clin Pharmacol Ther. 2018;104: 1155–1164. doi: 10.1002/cpt.1053 29484635

25. Lin J-W, Spaccapelo R, Schwarzer E, Sajid M, Annoura T, Deroost K, et al. Replication of Plasmodium in reticulocytes can occur without hemozoin formation, resulting in chloroquine resistance. J Exp Med. 2015;212: 893–903. doi: 10.1084/jem.20141731 25941254

26. Capuccini B, Lin J, Talavera-López C, Khan SM, Sodenkamp J, Spaccapelo R, et al. Transcriptomic profiling of microglia reveals signatures of cell activation and immune response, during experimental cerebral malaria. Sci Rep. 2016;6: 39258. doi: 10.1038/srep39258 27991544

27. Schwach F, Bushell E, Gomes AR, Anar B, Girling G, Herd C, et al. PlasmoGEM, a database supporting a community resource for large-scale experimental genetics in malaria parasites. Nucleic Acids Res. 2015;43: D1176–82. doi: 10.1093/nar/gku1143 25593348

28. Deu E, Yang Z, Wang F, Klemba M, Bogyo M. Use of activity-based probes to develop high throughput screening assays that can be performed in complex cell extracts. PLoS ONE. 2010;5: e11985. doi: 10.1371/journal.pone.0011985 20700487

29. Wang F, Krai P, Deu E, Bibb B, Lauritzen C, Pedersen J, et al. Biochemical characterization of Plasmodium falciparum dipeptidyl aminopeptidase 1. Mol Biochem Parasitol. 2011;175: 10–20. doi: 10.1016/j.molbiopara.2010.08.004 20833209

30. Deu E, de Vries LE, Sanchez MI, Groborz K, Kuppens L, Poreba M, et al. Characterization of P. falciparum dipeptidyl aminopeptidase 3 specificity identifies different amino acid preferences between peptide-based substrates and inhibitors. Preprint. Available from bioRxiv. 2018;1–32. doi: 10.1101/246124

31. Mycek MJ. Cathepsins. Methods in Enzymology. 1970;19: 285–315.

32. Horn M, Baudys M, Voburka Z, Kluh I, Vondrášek J, Mareš M. Free-thiol Cys331 exposed during activation process is critical for native tetramer structure of cathepsin C (dipeptidyl peptidase I). Protein Sci. 2002;11: 933–943. doi: 10.1110/ps.2910102 11910036

33. Arastu-Kapur S, Ponder EL, Fonović UP, Yeoh S, Yuan F, Fonović M, et al. Identification of proteases that regulate erythrocyte rupture by the malaria parasite Plasmodium falciparum. Nat Chem Biol. 2008;4: 203–213. doi: 10.1038/nchembio.70 18246061

34. Baell JB, Holloway GA. New substructure filters for removal of pan assay interference compounds (PAINS) from screening libraries and for their exclusion in bioassays. J Med Chem. 2010;53: 2719–2740. doi: 10.1021/jm901137j 20131845

35. Poreba M, Mihelic M, Krai P, Rajkovic J, Krezel A, Pawelczak M, et al. Unnatural amino acids increase activity and specificity of synthetic substrates for human and malarial cathepsin C. Amino Acids. 2014;46: 931–943. doi: 10.1007/s00726-013-1654-2 24381006

36. Marques AF, Gomes PSFC, Oliveira PL, Rosenthal PJ, Pascutti PG, Lima LMTR. Allosteric regulation of the Plasmodium falciparum cysteine protease falcipain-2 by heme. Arch Biochem Biophys. 2015;573: 92–99. doi: 10.1016/j.abb.2015.03.007 25791019

37. Marques AF, Esser D, Rosenthal PJ, Kassack MU, Lima LMTR. Falcipain-2 inhibition by suramin and suramin analogues. Bioorg Med Chem. 2013;21: 3667–3673. doi: 10.1016/j.bmc.2013.04.047 23680445

38. Bertoldo JB, Chiaradia-Delatorre LD, Mascarello A, Leal PC, Cordeiro MNS, Nunes RJ, et al. Synthetic compounds from an in house library as inhibitors of falcipain-2 from Plasmodium falciparum. J Enzyme Inhib Med Chem. 2015;30: 299–307. doi: 10.3109/14756366.2014.920839 24964346

39. Novinec M, Rebernik M, Lenarčič B. An allosteric site enables fine-tuning of cathepsin K by diverse effectors. FEBS Lett. 2016;590: 4507–4518. doi: 10.1002/1873-3468.12495 27859061

40. Methot N, Guay D, Rubin J, Ethier D, Ortega K, Wong S, et al. In vivo inhibition of serine protease processing requires a high fractional inhibition of cathepsin C. Mol Pharmacol. 2008;73: 1857–1865. doi: 10.1124/mol.108.045682 18326050

41. Methot N, Rubin J, Guay D, Beaulieu C, Ethier D, Reddy TJ, et al. Inhibition of the activation of multiple serine proteases with a cathepsin C inhibitor requires sustained exposure to prevent pro-enzyme processing. J Biol Chem. 2007;282: 20836–20846. doi: 10.1074/jbc.M702615200 17535802

42. Que X, Engel JC, Ferguson D, Wunderlich A, Tomavo S, Reed SL. Cathepsin Cs are key for the intracellular survival of the protozoan parasite, Toxoplasma gondii. J Biol Chem. 2007;282: 4994–5003. doi: 10.1074/jbc.M606764200 17164247


Článek vyšel v časopise

PLOS One


2019 Číslo 12
Nejčtenější tento týden
Nejčtenější v tomto čísle
Kurzy

Zvyšte si kvalifikaci online z pohodlí domova

KOST
Koncepce osteologické péče pro gynekology a praktické lékaře
nový kurz
Autoři: MUDr. František Šenk

Sekvenční léčba schizofrenie
Autoři: MUDr. Jana Hořínková

Hypertenze a hypercholesterolémie – synergický efekt léčby
Autoři: prof. MUDr. Hana Rosolová, DrSc.

Svět praktické medicíny 5/2023 (znalostní test z časopisu)

Imunopatologie? … a co my s tím???
Autoři: doc. MUDr. Helena Lahoda Brodská, Ph.D.

Všechny kurzy
Kurzy Podcasty Doporučená témata Časopisy
Přihlášení
Zapomenuté heslo

Zadejte e-mailovou adresu, se kterou jste vytvářel(a) účet, budou Vám na ni zaslány informace k nastavení nového hesla.

Přihlášení

Nemáte účet?  Registrujte se

#ADS_BOTTOM_SCRIPTS#