ABB  Vol.3 No.7 A , November 2012
Efficacy of 1-(N-acetyl-6-aminohexyl)-3-hydroxypyridin-4-one (CM1) in treatment of iron-loaded hepatocyte cultures
ABSTRACT
Excessive iron is toxic to cells and organelles, where it can generate harmful reactive oxygen species (ROS) resulting in oxidative tissue damage. Liver is the major organ for iron storage and redox active iron in this tissue can cause fibrosis and cirrhosis in β-thalassemia patients. Desferrioxamine (DFO), deferiprone (DFP) and deferasirox (DFX) are clinically approved iron chelators used for the treatment of patients with iron overload, but none of these chelators are completely free of side effects. In this study we report the properties of a new iron chelator 1-(N-acetyl-6-aminohexyl)-3-hydroxypyridin-4-one (CM1). The labile iron pool (LIP) content was measured by using a calcein fluorescence technique and the lipidperoxidation products were quantified using the thiobarbituric acid reactive substances (TBARS) method. The cytotoxicity of CM1 was also examined with the (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. CM1 was demonstrated to reduce iron-induced redox damage and to decrease the levels of the intracellular iron pool in hepatocytes. CM1 is a potentially useful iron-chelating agent which has potential to ameliorate liver iron overload and ROS-induced lipid peroxidation. CM1 is currently under investigation for oral efficacy.

Cite this paper
Pangjit, K. , Banjerdpongchai, R. , Phisalaphong, C. , Fucharoen, S. and Srichairatanakool, S. (2012) Efficacy of 1-(N-acetyl-6-aminohexyl)-3-hydroxypyridin-4-one (CM1) in treatment of iron-loaded hepatocyte cultures. Advances in Bioscience and Biotechnology, 3, 1060-1067. doi: 10.4236/abb.2012.327129.
References
[1]   Aisen, P., Enns, C. and Wessling-Resnick, M. (2001) Chemistry and biology of eukaryotic iron metabolism. The International Journal of Biochemistry and Cell Biology, 33, 940-959. doi:10.1016/S1357-2725(01)00063-2

[2]   Hershko, C. (2007) Iron loading and its clinical implications. American Journal of Hematology, 82, 1147-1148. doi:10.1002/ajh.21070

[3]   Emerit, J., Beaumont, C. and Trivin, F. (2001) Iron metabolism, free radicals, and oxidative injury. Biomedecine & Pharmacotherapy, 55, 333-339. doi:10.1016/S0753-3322(01)00068-3

[4]   Jensen, P.D. (2004) Evaluation of iron overload. British Journal of Haematology, 124, 697-711. doi:10.1111/j.1365-2141.2004.04838.x

[5]   Petrat, F., de Groot, H., Sustmann, R. and Rauen, U. (2002) The chelatable iron pool in living cells: A methodically defined quantity. Biological Chemistry, 383, 489-502. doi:10.1515/BC.2002.051

[6]   Staubli, A. and Boelsterli, U.A. (1998) The labile iron pool in hepatocytes: Prooxidant-induced increase in free iron precedes oxidative cell injury. American Journal of Physiology—Gastrointestinal and Liver Physiology, 274, 1031-1037.

[7]   Kakhlon, O. and Cabantchik, Z. I. (2002) The labile iron pool: Characterization, measurement, and participation in cellular processes. Free Radical Biology and Medicine, 33, 1037-1046. doi:10.1016/S0891-5849(02)01006-7

[8]   Gunther, T., Vormann, J. and Hollriegl, V. (1995) Effects of magnesium and iron on lipid peroxidation in cultured hepatocytes. Molecular Cell Biochemistry, 144, 141-145. doi:10.1007/BF00944393

[9]   Prus, E and Fibach, E. (2009) Effect of iron chelators on labile iron and oxidative status of thalassaemic erythroid cells. Acta Haematologica, 123, 14-20. doi:10.1159/000258958

[10]   Zanninelli, G., Loreal, O., Brissot, P., Konijn, A.M., Slotki, I.N., Hider, R.C. and Ioav Cabantchik, Z. (2002) The labile iron pool of hepatocytes in chronic and acute iron overload and chelator-induced iron deprivation. Journal of Hepatology, 36, 39-46. doi:10.1016/S0168-8278(01)00222-7

[11]   Brittenham, G.M. (2003) Iron chelators and iron toxicity. Alcohol, 30, 151-158. doi:10.1016/S0741-8329(03)00101-0

[12]   Neufeld, E.J. (2006) Oral chelators deferasirox and deferiprone for transfusional iron overload in thalassemia major: New data, new questions. Blood, 107, 3436-3441. doi:10.1182/blood-2006-02-002394

[13]   Porter, J.B. (2006) Deferasirox: An effective once-daily orally active iron chelator. Drugs Today (Barc), 42, 623-637. doi:10.1358/dot.2006.42.10.1009901

[14]   Hershko, C.M., Link, G.M., Konijn, A.M. and Cabantchik, Z.I. (2005) Iron chelation therapy. Current Hematology Reports, 4, 110-116.

[15]   Kwiatkowski, J.L. (2008) Oral iron chelators. Pediatric Clinics of North America, 55, 461-482. doi:10.1016/j.pcl.2008.01.005

[16]   Cappellini, M.D. and Piga, A. (2008) Current status in iron chelation in hemoglobinopathies. Current Molecular Medicine, 8, 663-674. doi:10.2174/156652408786241438

[17]   Porter, J.B. (1997) A risk-benefit assessment of iron-chelation therapy. Drug Safety, 17, 407-421. doi:10.2165/00002018-199717060-00006

[18]   Pangjit, K., Banjerdpongchai, R., Phisalaphong, C., Fucharoen, S., Xie, Y., Hider, R.C. and Srichairatanakool, S. Characterization and investigation of chelating activity of a novel oral iron chelator: 1-(N-acetyl-6-aminohexyl)-3-hydroxypyridin-4-one. European Journal of Pharmaceutical Sciences.

[19]   Chenoufi, N., Drenou, B., Loreal, O., Pigeon, C., Brissot, P. and Lescoat, G. (1998) Antiproliferative effect of deferiprone on the Hep G2 cell line. Biochemical Pharmacology, 56, 431-437. doi:10.1016/S0006-2952(98)00071-9

[20]   Baker, E., Baker, S.M. and Morgan, E.H. (1998) Characterisation of non-transferrin-bound iron (ferric citrate) uptake by rat hepatocytes in culture. Biochimica et Biophysica Acta, 1380, 21-30. doi:10.1016/S0304-4165(97)00120-7

[21]   Mosmann, T. (1983) Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assays. Journal of Immunological Methods, 65, 55-63. doi:10.1016/0022-1759(83)90303-4

[22]   Jagetia, G.C., Reddy, T.K., Venkatesha, V.A. and Kedlaya, R. (2004) Influence of naringin on ferric iron induced oxidative damage in vitro. Clinica Chimica Acta, 347, 189-197. doi:10.1016/j.cccn.2004.04.022

[23]   Huang, X., Dai, J., Fournier, J., Ali, A.M., Zhang, Q. and Frenkel, K. (2002) Ferrous ion autoxidation and its chelation in iron-loaded human liver Hep G2 cells. Free Radical Biology and Medicine, 32, 84-92. doi:10.1016/S0891-5849(01)00770-5

[24]   Tenopoulou, M., Kurz, T., Doulias, P.-T., Galaris D. and Brunk, U. T. (2007) Does the calcein-AM method assay the total cellular ‘labile iron pool’ or only a fraction of it? Biochem Journal, 15, 261-266.

[25]   Heyduk, T., Ma, Y., Tang, H. and Ebright, R.H. (1996) Fluorescence anisotropy: rapid, quantitative assay for protein-DNA and protein-protein interaction. Methods in Enzymology, 274, 492-503. doi:10.1016/S0076-6879(96)74039-9

[26]   Sturm, B., Goldenberg, H. and Scheiber-Mojdehkar, B. (2003) Transient increase of the labile iron pool in HepG2 cells by intravenous iron preparations. European Journal of Biochemistry, 270, 3731-3738.

[27]   Glickstein, H., Breuer, W., Loyevsky, M., Konijn, A.M., Shanzer, A. and Cabantchik, Z.I. (1996) Differential cytotoxicity of iron chelators on malaria-infected cells versus mammalian cells. Blood, 87, 4871-4878.

[28]   Porter, J.B., Gyparaki, M., Burke, L.C., Huehns, E.R., Sarpong, P., Saez, V. and Hider, R.C. (1988) Iron mobilization from hepatocyte monolayer cultures by chelators: the importance of membrane permeability and the iron-binding constant. Blood, 72, 1497-1503.

[29]   Selden, C., Owen, M., Hopkins, J.M. and Peters, T.J. (1980) Studies on the concentration and intracellular localization of iron proteins in liver biopsy specimens from patients with iron overload with special reference to their role in lysosomal disruption. British Journal of Haematology, 44, 593-603. doi:10.1111/j.1365-2141.1980.tb08714.x

[30]   Wright, T.L., Brissot, P., Ma, W.L. and Weisiger, R.A. (1986) Characterization of non-transferrin-bound iron clearance by rat liver. Journal of Biological Chemistry 261, 10909-10914.

[31]   Brissot, P., Wright, T.L., Ma, W.L. and Weisiger, R.A. (1985) Efficient clearance of non-transferrin-bound iron by rat liver. Implications for hepatic iron loading in iron overload states. Journal of Clinical Investigation, 76, 1463-1470. doi:10.1172/JCI112125

[32]   Halliwell, B. and Gutteridge, J.M. (1986) Oxygen free radicals and iron in relation to biology and medicine: some problems and concepts. Archives of Biochemistry and Biophysics, 246, 501-514. doi:10.1016/0003-9861(86)90305-X

[33]   Turlin, B. and Deugnier, Y. (2002) Iron overload disorders. Clinical Liver Disease, 6, 481-496. doi:10.1016/S1089-3261(02)00004-1

[34]   Kakhlon, O., Gruenbaum, Y. and Cabantchik, Z.I. (2001) Repression of ferritin expression increases the labile iron pool, oxidative stress, and short-term growth of human erythroleukemia cells. Blood, 97, 2863-2871. doi:10.1182/blood.V97.9.2863

[35]   Breuer, W., Epsztejn, S. and Cabantchik, Z.I. (1996) Dynamics of the cytosolic chelatable iron pool of K562 cells. FEBS Letters, 382, 304-308. doi:10.1016/0014-5793(96)00190-1

[36]   Parkes, J.G., Randell, E.W., Olivieri, N.F. and Templeton, D.M. (1995) Modulation by iron loading and chelation of the uptake of non-transferrin-bound iron by human liver cells. Biochimica et Biophysica Acta, 1243, 373-380. doi:10.1016/0304-4165(94)00162-Q

 
 
Top