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. 1999 Nov;81(6):981–988. doi: 10.1038/sj.bjc.6690796

N-substituted benzamides inhibit NFκB activation and induce apoptosis by separate mechanisms

D Liberg 1, B Lazarevic 1, R W Pero 2, T Leanderson 1
PMCID: PMC2362952  PMID: 10576654

Abstract

Benzamides have been in clinical use for many years in treatment against various disorders. A recent application is that as a sensitizer for radio- or chemotherapies. We have here analysed the mechanism of action of N-substituted benzamides using an in vitro system. We found that while procainamide was biologically inert in our system, the addition of a chloride in the 3′ position of the benzamide ring created a compound (declopramide) that induced rapid apoptosis. Furthermore, declopramide also inhibited NFκB activation by inhibition of IκBβ breakdown. An acetylated variant of declopramide, N-acetyl declopramide, showed no effect with regard to rapid apoptosis induction but was a potent inhibitor of NFκB activation. In fact, the addition of an acetyl group to procainamide in the 4′ position was sufficient to convert this biologically inactive substance to a potent inhibitor of NFκB activation. These findings suggest two potential mechanisms, induction of early apoptosis and inhibition of NFκB mediated salvage from apoptosis, for the biological effect of N-substituted benzamides as radio- and chemo-sensitizers. In addition it suggests that N-substituted benzamides are potential candidates for the development of anti-inflammatory compounds using NFκB as a drug target. © 1999 Cancer Research Campaign

Keywords: benzamides, apoptosis, NFκB

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Selected References

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  1. Baeuerle P. A., Baltimore D. NF-kappa B: ten years after. Cell. 1996 Oct 4;87(1):13–20. doi: 10.1016/s0092-8674(00)81318-5. [DOI] [PubMed] [Google Scholar]
  2. Baldwin A. S., Jr The NF-kappa B and I kappa B proteins: new discoveries and insights. Annu Rev Immunol. 1996;14:649–683. doi: 10.1146/annurev.immunol.14.1.649. [DOI] [PubMed] [Google Scholar]
  3. Beg A. A., Baltimore D. An essential role for NF-kappaB in preventing TNF-alpha-induced cell death. Science. 1996 Nov 1;274(5288):782–784. doi: 10.1126/science.274.5288.782. [DOI] [PubMed] [Google Scholar]
  4. Chen Z. J., Parent L., Maniatis T. Site-specific phosphorylation of IkappaBalpha by a novel ubiquitination-dependent protein kinase activity. Cell. 1996 Mar 22;84(6):853–862. doi: 10.1016/s0092-8674(00)81064-8. [DOI] [PubMed] [Google Scholar]
  5. Cohen G. M. Caspases: the executioners of apoptosis. Biochem J. 1997 Aug 15;326(Pt 1):1–16. doi: 10.1042/bj3260001. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. DiDonato J., Mercurio F., Rosette C., Wu-Li J., Suyang H., Ghosh S., Karin M. Mapping of the inducible IkappaB phosphorylation sites that signal its ubiquitination and degradation. Mol Cell Biol. 1996 Apr;16(4):1295–1304. doi: 10.1128/mcb.16.4.1295. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dolle R. E., Hoyer D., Prasad C. V., Schmidt S. J., Helaszek C. T., Miller R. E., Ator M. A. P1 aspartate-based peptide alpha-((2,6-dichlorobenzoyl)oxy)methyl ketones as potent time-dependent inhibitors of interleukin-1 beta-converting enzyme. J Med Chem. 1994 Mar 4;37(5):563–564. doi: 10.1021/jm00031a003. [DOI] [PubMed] [Google Scholar]
  8. Enari M., Sakahira H., Yokoyama H., Okawa K., Iwamatsu A., Nagata S. A caspase-activated DNase that degrades DNA during apoptosis, and its inhibitor ICAD. Nature. 1998 Jan 1;391(6662):43–50. doi: 10.1038/34112. [DOI] [PubMed] [Google Scholar]
  9. Ghosh S., May M. J., Kopp E. B. NF-kappa B and Rel proteins: evolutionarily conserved mediators of immune responses. Annu Rev Immunol. 1998;16:225–260. doi: 10.1146/annurev.immunol.16.1.225. [DOI] [PubMed] [Google Scholar]
  10. Harrington R. A., Hamilton C. W., Brogden R. N., Linkewich J. A., Romankiewicz J. A., Heel R. C. Metoclopramide. An updated review of its pharmacological properties and clinical use. Drugs. 1983 May;25(5):451–494. doi: 10.2165/00003495-198325050-00002. [DOI] [PubMed] [Google Scholar]
  11. Hua J., Olsson A. R., Pero R. W. Neutral metoclopramide sensitizes cytotoxicity induced by ionizing radiation in SCID mice xenografted with a human brain astrocytoma. Int J Cancer. 1997 Dec 10;73(6):871–874. doi: 10.1002/(sici)1097-0215(19971210)73:6<871::aid-ijc18>3.0.co;2-7. [DOI] [PubMed] [Google Scholar]
  12. Hua J., Olsson A., Sheng Y., Pero R. W. Acidic and neutralized metoclopramide formulations sensitize ionizing radiation induced cytotoxicity in a human lung adenocarcinoma xenografted to scid mice. Anticancer Drugs. 1995 Jun;6(3):451–455. doi: 10.1097/00001813-199506000-00014. [DOI] [PubMed] [Google Scholar]
  13. Hua J., Pero R. W. Toxicity, antitumor and chemosensitizing effects of 3-chloroprocainamide. Acta Oncol. 1997;36(8):811–816. doi: 10.3109/02841869709001362. [DOI] [PubMed] [Google Scholar]
  14. Kjellén E., Pero R. W., Brun E., Ewers S. B., Jarlman O., Knös T., Malmström P., Tennvall J., Killander D., Olsson A. A phase I/II evaluation of metoclopramide as a radiosensitiser in patients with inoperable squamous cell carcinoma of the lung. Eur J Cancer. 1995 Dec;31A(13-14):2196–2202. doi: 10.1016/0959-8049(95)00424-6. [DOI] [PubMed] [Google Scholar]
  15. Kjellén E., Wennerberg J., Pero R. Metoclopramide enhances the effect of cisplatin on xenografted squamous cell carcinoma of the head and neck. Br J Cancer. 1989 Feb;59(2):247–250. doi: 10.1038/bjc.1989.50. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Koopman G., Reutelingsperger C. P., Kuijten G. A., Keehnen R. M., Pals S. T., van Oers M. H. Annexin V for flow cytometric detection of phosphatidylserine expression on B cells undergoing apoptosis. Blood. 1994 Sep 1;84(5):1415–1420. [PubMed] [Google Scholar]
  17. Le Bail O., Schmidt-Ullrich R., Israël A. Promoter analysis of the gene encoding the I kappa B-alpha/MAD3 inhibitor of NF-kappa B: positive regulation by members of the rel/NF-kappa B family. EMBO J. 1993 Dec 15;12(13):5043–5049. doi: 10.1002/j.1460-2075.1993.tb06197.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Lee F. S., Hagler J., Chen Z. J., Maniatis T. Activation of the IkappaB alpha kinase complex by MEKK1, a kinase of the JNK pathway. Cell. 1997 Jan 24;88(2):213–222. doi: 10.1016/s0092-8674(00)81842-5. [DOI] [PubMed] [Google Scholar]
  19. Liberg D., Sigvardsson M., Bemark M., Leanderson T. Differentiation-specific, octamer-dependent costimulation of kappa transcription. J Immunol. 1998 Apr 15;160(8):3899–3907. [PubMed] [Google Scholar]
  20. Liu Z. G., Hsu H., Goeddel D. V., Karin M. Dissection of TNF receptor 1 effector functions: JNK activation is not linked to apoptosis while NF-kappaB activation prevents cell death. Cell. 1996 Nov 1;87(3):565–576. doi: 10.1016/s0092-8674(00)81375-6. [DOI] [PubMed] [Google Scholar]
  21. Lybak S., Kjellén E., Wennerberg J., Pero R. Metoclopramide enhances the effect of ionizing radiation on xenografted squamous cell carcinoma of the head and neck. Int J Radiat Oncol Biol Phys. 1990 Dec;19(6):1419–1424. doi: 10.1016/0360-3016(90)90353-l. [DOI] [PubMed] [Google Scholar]
  22. Lybak S., Pero R. W. The benzamide derivative metoclopramide causes DNA damage and inhibition of DNA repair in human peripheral mononuclear leukocytes at clinically relevant doses. Carcinogenesis. 1991 Sep;12(9):1613–1617. doi: 10.1093/carcin/12.9.1613. [DOI] [PubMed] [Google Scholar]
  23. Olsson A. R., Hua J., Sheng Y., Pero R. W. Neutral metoclopramide induces tumor cytotoxicity and sensitizes ionizing radiation of a human lung adenocarcinoma and virus induced sarcoma in mice. Acta Oncol. 1997;36(3):323–330. doi: 10.3109/02841869709001271. [DOI] [PubMed] [Google Scholar]
  24. Paige C. J., Kincade P. W., Ralph P. Murine B cell leukemia line with inducible surface immunoglobulin expression. J Immunol. 1978 Aug;121(2):641–647. [PubMed] [Google Scholar]
  25. Pero R. W., Axelsson B., Siemann D., Chaplin D., Dougherty G. Newly discovered anti-inflammatory properties of the benzamides and nicotinamides. Mol Cell Biochem. 1999 Mar;193(1-2):119–125. [PubMed] [Google Scholar]
  26. Pero R. W., Olsson A., Amiri A., Chaplin D. Multiple mechanisms of action of the benzamides and nicotinamides as sensitizers of radiotherapy: opportunities for drug design. Cancer Detect Prev. 1998;22(3):225–236. doi: 10.1046/j.1525-1500.1998.0oa30.x. [DOI] [PubMed] [Google Scholar]
  27. Perry R. P., Kelley D. E. Immunoglobulin messenger RNAs in murine cell lines that have characteristics of immature B lymphocytes. Cell. 1979 Dec;18(4):1333–1339. doi: 10.1016/0092-8674(79)90243-5. [DOI] [PubMed] [Google Scholar]
  28. Philpott N. J., Turner A. J., Scopes J., Westby M., Marsh J. C., Gordon-Smith E. C., Dalgleish A. G., Gibson F. M. The use of 7-amino actinomycin D in identifying apoptosis: simplicity of use and broad spectrum of application compared with other techniques. Blood. 1996 Mar 15;87(6):2244–2251. [PubMed] [Google Scholar]
  29. Rang H. P., Urban L. New molecules in analgesia. Br J Anaesth. 1995 Aug;75(2):145–156. doi: 10.1093/bja/75.2.145. [DOI] [PubMed] [Google Scholar]
  30. Rosoff P. M., Cantley L. C. Lipopolysaccharide and phorbol esters induce differentiation but have opposite effects on phosphatidylinositol turnover and Ca2+ mobilization in 70Z/3 pre-B lymphocytes. J Biol Chem. 1985 Aug 5;260(16):9209–9215. [PubMed] [Google Scholar]
  31. Rosoff P. M., Stein L. F., Cantley L. C. Phorbol esters induce differentiation in a pre-B-lymphocyte cell line by enhancing Na+/H+ exchange. J Biol Chem. 1984 Jun 10;259(11):7056–7060. [PubMed] [Google Scholar]
  32. Sakahira H., Enari M., Nagata S. Cleavage of CAD inhibitor in CAD activation and DNA degradation during apoptosis. Nature. 1998 Jan 1;391(6662):96–99. doi: 10.1038/34214. [DOI] [PubMed] [Google Scholar]
  33. Schmid I., Uittenbogaart C. H., Keld B., Giorgi J. V. A rapid method for measuring apoptosis and dual-color immunofluorescence by single laser flow cytometry. J Immunol Methods. 1994 Apr 15;170(2):145–157. doi: 10.1016/0022-1759(94)90390-5. [DOI] [PubMed] [Google Scholar]
  34. Schreiber E., Matthias P., Müller M. M., Schaffner W. Rapid detection of octamer binding proteins with 'mini-extracts', prepared from a small number of cells. Nucleic Acids Res. 1989 Aug 11;17(15):6419–6419. doi: 10.1093/nar/17.15.6419. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Sen R., Baltimore D. Inducibility of kappa immunoglobulin enhancer-binding protein Nf-kappa B by a posttranslational mechanism. Cell. 1986 Dec 26;47(6):921–928. doi: 10.1016/0092-8674(86)90807-x. [DOI] [PubMed] [Google Scholar]
  36. Thornberry N. A., Bull H. G., Calaycay J. R., Chapman K. T., Howard A. D., Kostura M. J., Miller D. K., Molineaux S. M., Weidner J. R., Aunins J. A novel heterodimeric cysteine protease is required for interleukin-1 beta processing in monocytes. Nature. 1992 Apr 30;356(6372):768–774. doi: 10.1038/356768a0. [DOI] [PubMed] [Google Scholar]
  37. Van Antwerp D. J., Martin S. J., Kafri T., Green D. R., Verma I. M. Suppression of TNF-alpha-induced apoptosis by NF-kappaB. Science. 1996 Nov 1;274(5288):787–789. doi: 10.1126/science.274.5288.787. [DOI] [PubMed] [Google Scholar]
  38. Wang C. Y., Mayo M. W., Baldwin A. S., Jr TNF- and cancer therapy-induced apoptosis: potentiation by inhibition of NF-kappaB. Science. 1996 Nov 1;274(5288):784–787. doi: 10.1126/science.274.5288.784. [DOI] [PubMed] [Google Scholar]
  39. Werning J. W., Stepnick D. W., Jafri A., Megerian C. A., Antunez A. R., Zaidi S. I. Metoclopramide enhances the effect of photodynamic therapy on xenografted human squamous cell carcinoma of the head and neck. Arch Otolaryngol Head Neck Surg. 1995 Jul;121(7):783–789. doi: 10.1001/archotol.1995.01890070069015. [DOI] [PubMed] [Google Scholar]
  40. Whiteside S. T., Epinat J. C., Rice N. R., Israël A. I kappa B epsilon, a novel member of the I kappa B family, controls RelA and cRel NF-kappa B activity. EMBO J. 1997 Mar 17;16(6):1413–1426. doi: 10.1093/emboj/16.6.1413. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Whiteside S. T., Israël A. I kappa B proteins: structure, function and regulation. Semin Cancer Biol. 1997 Apr;8(2):75–82. doi: 10.1006/scbi.1997.0058. [DOI] [PubMed] [Google Scholar]
  42. Wu M., Lee H., Bellas R. E., Schauer S. L., Arsura M., Katz D., FitzGerald M. J., Rothstein T. L., Sherr D. H., Sonenshein G. E. Inhibition of NF-kappaB/Rel induces apoptosis of murine B cells. EMBO J. 1996 Sep 2;15(17):4682–4690. [PMC free article] [PubMed] [Google Scholar]
  43. de Martin R., Vanhove B., Cheng Q., Hofer E., Csizmadia V., Winkler H., Bach F. H. Cytokine-inducible expression in endothelial cells of an I kappa B alpha-like gene is regulated by NF kappa B. EMBO J. 1993 Jul;12(7):2773–2779. doi: 10.1002/j.1460-2075.1993.tb05938.x. [DOI] [PMC free article] [PubMed] [Google Scholar]

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