Skip to main content
PMC home page
Molecular Medicine logoLink to Molecular Medicine
. 1999 Sep;5(9):585–594.

Peptidomimetic fluoromethylketone rescues mice from lethal endotoxic shock.

S R Grobmyer 1, R C Armstrong 1, S C Nicholson 1, C Gabay 1, W P Arend 1, S H Potter 1, M Melchior 1, L C Fritz 1, C F Nathan 1
PMCID: PMC2230465  PMID: 10551900

Abstract

BACKGROUND: Septic shock is a leading cause of mortality in intensive care units. No new interventions in the last 20 years have made a substantial impact on the outcome of patients with septic shock. Identification of inhibitable pathways that mediate death in shock is an important goal. MATERIALS AND METHODS: Two novel caspase inhibitors, (2-indolyl)-carbonyl-Ala-Asp-fluoromethylketone (IDN 1529) and (1-methyl-3-methyl-2-indolyl)-carbonyl-Val-Asp-fluoromethylketone (IDN 1965), were studied in a murine model of endotoxic shock. RESULTS: IDN 1529 prolonged survival when given before or up to 3 hr after high-dose LPS (p < 0.01) and increased by 2.2-fold the number of animals surviving longterm after a lower dose of LPS (p < 0.01). Despite its similar chemical structure, IDN 1965 lacked these protective effects. Both compounds inhibited caspases 1, 2, 3, 6, 8, and 9, and both afforded comparable reduction in Fas- and LPS-induced caspase 3-like activity and apoptosis. Paradoxically, administration of IDN 1529 but not IDN 1965 led to an increase in the LPS-induced elevation of serum cytokines related directly (IL-1beta, IL-18) or indirectly (IL-1alpha, IL-1Ra) to the action of caspase 1. CONCLUSIONS: A process that appears to be distinct from both apoptosis and the release of inflammatory cytokines is a late-acting requirement for lethality in endotoxic shock. Inhibition of this process can rescue mice even when therapy is initiated after LPS has made the mice severely ill.

Full text

PDF

Images in this article

590Fig. 4
on p.590
591Fig. 5
on p.591

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Armstrong R. C., Aja T., Xiang J., Gaur S., Krebs J. F., Hoang K., Bai X., Korsmeyer S. J., Karanewsky D. S., Fritz L. C. Fas-induced activation of the cell death-related protease CPP32 Is inhibited by Bcl-2 and by ICE family protease inhibitors. J Biol Chem. 1996 Jul 12;271(28):16850–16855. doi: 10.1074/jbc.271.28.16850. [DOI] [PubMed] [Google Scholar]
  2. Astiz M. E., Rackow E. C. Septic shock. Lancet. 1998 May 16;351(9114):1501–1505. doi: 10.1016/S0140-6736(98)01134-9. [DOI] [PubMed] [Google Scholar]
  3. Beutler B., Milsark I. W., Cerami A. C. Passive immunization against cachectin/tumor necrosis factor protects mice from lethal effect of endotoxin. Science. 1985 Aug 30;229(4716):869–871. doi: 10.1126/science.3895437. [DOI] [PubMed] [Google Scholar]
  4. Black R. A., Kronheim S. R., Cantrell M., Deeley M. C., March C. J., Prickett K. S., Wignall J., Conlon P. J., Cosman D., Hopp T. P. Generation of biologically active interleukin-1 beta by proteolytic cleavage of the inactive precursor. J Biol Chem. 1988 Jul 5;263(19):9437–9442. [PubMed] [Google Scholar]
  5. Bohlinger I., Leist M., Gantner F., Angermüller S., Tiegs G., Wendel A. DNA fragmentation in mouse organs during endotoxic shock. Am J Pathol. 1996 Oct;149(4):1381–1393. [PMC free article] [PubMed] [Google Scholar]
  6. Casals-Stenzel J. Protective effect of WEB 2086, a novel antagonist of platelet activating factor, in endotoxin shock. Eur J Pharmacol. 1987 Mar 17;135(2):117–122. doi: 10.1016/0014-2999(87)90602-9. [DOI] [PubMed] [Google Scholar]
  7. Dinarello C. A. Interleukin-1. Cytokine Growth Factor Rev. 1997 Dec;8(4):253–265. doi: 10.1016/s1359-6101(97)00023-3. [DOI] [PubMed] [Google Scholar]
  8. Dinarello C. A., Novick D., Puren A. J., Fantuzzi G., Shapiro L., Mühl H., Yoon D. Y., Reznikov L. L., Kim S. H., Rubinstein M. Overview of interleukin-18: more than an interferon-gamma inducing factor. J Leukoc Biol. 1998 Jun;63(6):658–664. [PubMed] [Google Scholar]
  9. Doherty G. M., Lange J. R., Langstein H. N., Alexander H. R., Buresh C. M., Norton J. A. Evidence for IFN-gamma as a mediator of the lethality of endotoxin and tumor necrosis factor-alpha. J Immunol. 1992 Sep 1;149(5):1666–1670. [PubMed] [Google Scholar]
  10. Fletcher D. S., Agarwal L., Chapman K. T., Chin J., Egger L. A., Limjuco G., Luell S., MacIntyre D. E., Peterson E. P., Thornberry N. A. A synthetic inhibitor of interleukin-1 beta converting enzyme prevents endotoxin-induced interleukin-1 beta production in vitro and in vivo. J Interferon Cytokine Res. 1995 Mar;15(3):243–248. doi: 10.1089/jir.1995.15.243. [DOI] [PubMed] [Google Scholar]
  11. Gabay C., Porter B., Fantuzzi G., Arend W. P. Mouse IL-1 receptor antagonist isoforms: complementary DNA cloning and protein expression of intracellular isoform and tissue distribution of secreted and intracellular IL-1 receptor antagonist in vivo. J Immunol. 1997 Dec 15;159(12):5905–5913. [PubMed] [Google Scholar]
  12. Gabay C., Smith M. F., Eidlen D., Arend W. P. Interleukin 1 receptor antagonist (IL-1Ra) is an acute-phase protein. J Clin Invest. 1997 Jun 15;99(12):2930–2940. doi: 10.1172/JCI119488. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Ghayur T., Banerjee S., Hugunin M., Butler D., Herzog L., Carter A., Quintal L., Sekut L., Talanian R., Paskind M. Caspase-1 processes IFN-gamma-inducing factor and regulates LPS-induced IFN-gamma production. Nature. 1997 Apr 10;386(6625):619–623. doi: 10.1038/386619a0. [DOI] [PubMed] [Google Scholar]
  14. Gu Y., Kuida K., Tsutsui H., Ku G., Hsiao K., Fleming M. A., Hayashi N., Higashino K., Okamura H., Nakanishi K. Activation of interferon-gamma inducing factor mediated by interleukin-1beta converting enzyme. Science. 1997 Jan 10;275(5297):206–209. doi: 10.1126/science.275.5297.206. [DOI] [PubMed] [Google Scholar]
  15. Guirao X., Lowry S. F. Biologic control of injury and inflammation: much more than too little or too late. World J Surg. 1996 May;20(4):437–446. doi: 10.1007/s002689900069. [DOI] [PubMed] [Google Scholar]
  16. Haendeler J., Messmer U. K., Brüne B., Neugebauer E., Dimmeler S. Endotoxic shock leads to apoptosis in vivo and reduces Bcl-2. Shock. 1996 Dec;6(6):405–409. doi: 10.1097/00024382-199612000-00004. [DOI] [PubMed] [Google Scholar]
  17. Haimovitz-Friedman A., Cordon-Cardo C., Bayoumy S., Garzotto M., McLoughlin M., Gallily R., Edwards C. K., 3rd, Schuchman E. H., Fuks Z., Kolesnick R. Lipopolysaccharide induces disseminated endothelial apoptosis requiring ceramide generation. J Exp Med. 1997 Dec 1;186(11):1831–1841. doi: 10.1084/jem.186.11.1831. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hazuda D. J., Strickler J., Kueppers F., Simon P. L., Young P. R. Processing of precursor interleukin 1 beta and inflammatory disease. J Biol Chem. 1990 Apr 15;265(11):6318–6322. [PubMed] [Google Scholar]
  19. Hirsch E., Irikura V. M., Paul S. M., Hirsh D. Functions of interleukin 1 receptor antagonist in gene knockout and overproducing mice. Proc Natl Acad Sci U S A. 1996 Oct 1;93(20):11008–11013. doi: 10.1073/pnas.93.20.11008. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Hogquist K. A., Nett M. A., Unanue E. R., Chaplin D. D. Interleukin 1 is processed and released during apoptosis. Proc Natl Acad Sci U S A. 1991 Oct 1;88(19):8485–8489. doi: 10.1073/pnas.88.19.8485. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Irmler M., Hertig S., MacDonald H. R., Sadoul R., Becherer J. D., Proudfoot A., Solari R., Tschopp J. Granzyme A is an interleukin 1 beta-converting enzyme. J Exp Med. 1995 May 1;181(5):1917–1922. doi: 10.1084/jem.181.5.1917. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Jaeschke H., Fisher M. A., Lawson J. A., Simmons C. A., Farhood A., Jones D. A. Activation of caspase 3 (CPP32)-like proteases is essential for TNF-alpha-induced hepatic parenchymal cell apoptosis and neutrophil-mediated necrosis in a murine endotoxin shock model. J Immunol. 1998 Apr 1;160(7):3480–3486. [PubMed] [Google Scholar]
  23. Krege J. H., Hodgin J. B., Hagaman J. R., Smithies O. A noninvasive computerized tail-cuff system for measuring blood pressure in mice. Hypertension. 1995 May;25(5):1111–1115. doi: 10.1161/01.hyp.25.5.1111. [DOI] [PubMed] [Google Scholar]
  24. Kuida K., Lippke J. A., Ku G., Harding M. W., Livingston D. J., Su M. S., Flavell R. A. Altered cytokine export and apoptosis in mice deficient in interleukin-1 beta converting enzyme. Science. 1995 Mar 31;267(5206):2000–2003. doi: 10.1126/science.7535475. [DOI] [PubMed] [Google Scholar]
  25. Li P., Allen H., Banerjee S., Franklin S., Herzog L., Johnston C., McDowell J., Paskind M., Rodman L., Salfeld J. Mice deficient in IL-1 beta-converting enzyme are defective in production of mature IL-1 beta and resistant to endotoxic shock. Cell. 1995 Feb 10;80(3):401–411. doi: 10.1016/0092-8674(95)90490-5. [DOI] [PubMed] [Google Scholar]
  26. MacMicking J. D., Nathan C., Hom G., Chartrain N., Fletcher D. S., Trumbauer M., Stevens K., Xie Q. W., Sokol K., Hutchinson N. Altered responses to bacterial infection and endotoxic shock in mice lacking inducible nitric oxide synthase. Cell. 1995 May 19;81(4):641–650. doi: 10.1016/0092-8674(95)90085-3. [DOI] [PubMed] [Google Scholar]
  27. Matsushima K., Taguchi M., Kovacs E. J., Young H. A., Oppenheim J. J. Intracellular localization of human monocyte associated interleukin 1 (IL 1) activity and release of biologically active IL 1 from monocytes by trypsin and plasmin. J Immunol. 1986 Apr 15;136(8):2883–2891. [PubMed] [Google Scholar]
  28. McCabe W. R. Immunization with R mutants of S. Minnesota. I. Protection against challenge with heterologous gram-negative bacilli. J Immunol. 1972 Mar;108(3):601–610. [PubMed] [Google Scholar]
  29. Mignon A., Rouquet N., Fabre M., Martin S., Pagès J. C., Dhainaut J. F., Kahn A., Briand P., Joulin V. LPS challenge in D-galactosamine-sensitized mice accounts for caspase-dependent fulminant hepatitis, not for septic shock. Am J Respir Crit Care Med. 1999 Apr;159(4 Pt 1):1308–1315. doi: 10.1164/ajrccm.159.4.9712012. [DOI] [PubMed] [Google Scholar]
  30. Miller B. E., Krasney P. A., Gauvin D. M., Holbrook K. B., Koonz D. J., Abruzzese R. V., Miller R. E., Pagani K. A., Dolle R. E., Ator M. A. Inhibition of mature IL-1 beta production in murine macrophages and a murine model of inflammation by WIN 67694, an inhibitor of IL-1 beta converting enzyme. J Immunol. 1995 Feb 1;154(3):1331–1338. [PubMed] [Google Scholar]
  31. Mittl P. R., Di Marco S., Krebs J. F., Bai X., Karanewsky D. S., Priestle J. P., Tomaselli K. J., Grütter M. G. Structure of recombinant human CPP32 in complex with the tetrapeptide acetyl-Asp-Val-Ala-Asp fluoromethyl ketone. J Biol Chem. 1997 Mar 7;272(10):6539–6547. doi: 10.1074/jbc.272.10.6539. [DOI] [PubMed] [Google Scholar]
  32. Perregaux D. G., Gabel C. A. Post-translational processing of murine IL-1: evidence that ATP-induced release of IL-1 alpha and IL-1 beta occurs via a similar mechanism. J Immunol. 1998 Mar 1;160(5):2469–2477. [PubMed] [Google Scholar]
  33. Perregaux D., Barberia J., Lanzetti A. J., Geoghegan K. F., Carty T. J., Gabel C. A. IL-1 beta maturation: evidence that mature cytokine formation can be induced specifically by nigericin. J Immunol. 1992 Aug 15;149(4):1294–1303. [PubMed] [Google Scholar]
  34. Puren A. J., Fantuzzi G., Dinarello C. A. Gene expression, synthesis, and secretion of interleukin 18 and interleukin 1beta are differentially regulated in human blood mononuclear cells and mouse spleen cells. Proc Natl Acad Sci U S A. 1999 Mar 2;96(5):2256–2261. doi: 10.1073/pnas.96.5.2256. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Salvesen G. S., Dixit V. M. Caspases: intracellular signaling by proteolysis. Cell. 1997 Nov 14;91(4):443–446. doi: 10.1016/s0092-8674(00)80430-4. [DOI] [PubMed] [Google Scholar]
  36. Schimke J., Mathison J., Morgiewicz J., Ulevitch R. J. Anti-CD14 mAb treatment provides therapeutic benefit after in vivo exposure to endotoxin. Proc Natl Acad Sci U S A. 1998 Nov 10;95(23):13875–13880. doi: 10.1073/pnas.95.23.13875. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Schotte P., Declercq W., Van Huffel S., Vandenabeele P., Beyaert R. Non-specific effects of methyl ketone peptide inhibitors of caspases. FEBS Lett. 1999 Jan 8;442(1):117–121. doi: 10.1016/s0014-5793(98)01640-8. [DOI] [PubMed] [Google Scholar]
  38. Schönbeck U., Mach F., Libby P. Generation of biologically active IL-1 beta by matrix metalloproteinases: a novel caspase-1-independent pathway of IL-1 beta processing. J Immunol. 1998 Oct 1;161(7):3340–3346. [PubMed] [Google Scholar]
  39. Silver G. M., Gamelli R. L., O'Reilly M., Hebert J. C. The effect of interleukin 1 alpha on survival in a murine model of burn wound sepsis. Arch Surg. 1990 Jul;125(7):922–925. doi: 10.1001/archsurg.1990.01410190120020. [DOI] [PubMed] [Google Scholar]
  40. Thornberry N. A., Lazebnik Y. Caspases: enemies within. Science. 1998 Aug 28;281(5381):1312–1316. doi: 10.1126/science.281.5381.1312. [DOI] [PubMed] [Google Scholar]
  41. Wang S., Miura M., Jung Y. K., Zhu H., Li E., Yuan J. Murine caspase-11, an ICE-interacting protease, is essential for the activation of ICE. Cell. 1998 Feb 20;92(4):501–509. doi: 10.1016/s0092-8674(00)80943-5. [DOI] [PubMed] [Google Scholar]

Articles from Molecular Medicine are provided here courtesy of The Feinstein Institute for Medical Research at North Shore LIJ

RESOURCES