Summary
The vast majority of preclinical studies of HDAC inhibitors (HDAC-I) focus on the drug–target (cancer) cell interaction, whereas little attention is paid to the effects on non-target healthy cells, which could provide decisive information to eliminate potential cytotoxic compounds at a very early stage during drug development. In the current study we used cultures of primary rat hepatocytes as a read out system to select for the most potent HDAC-I in the group of structural analogues of an archetypal HDAC-I, namely Trichostatin A. This kind of approach allowed selecting compounds with high biological activity and with no apparent toxicity towards cultured hepatocytes.
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References
Glaser KB (2007) HDAC inhibitors: clinical update and mechanism-based potential. Biochem Pharmacol 74:659–671 doi:10.1016/j.bcp.2007.04.007
Bolden JE, Peart MJ, Johnstone RW (2006) Anticancer activities of histone deacetylase inhibitors. Nat Rev Drug Discov 5:769–784 doi:10.1038/nrd2133
Glozak MA, Seto E (2007) Histone deacetylases and cancer. Oncogene 26:5420–5432 doi:10.1038/sj.onc.1210610
Huang L (2006) Targeting histone deacetylases for the treatment of cancer and inflammatory diseases. J Cell Physiol 209:611–616 doi:10.1002/jcp.20781
Hildmann C, Riester D, Schwienhorst A (2007) Histone deacetylases—an important class of cellular regulators with a variety of functions. Appl Microbiol Biotechnol 75:487–497 doi:10.1007/s00253-007-0911-2
Papeleu P, Vanhaecke T, Elaut G et al (2005) Differential effects of histone deacetylase inhibitors in tumor and normal cells-what is the toxicological relevance. Crit Rev Toxicol 35:363–378 doi:10.1080/10408440590935639
Marcos R, Monteiro RA, Rocha E (2006) Design-based stereological estimation of hepatocyte number, by combining the smooth optical fractionator and immunocytochemistry with anti-carcinoembryonic antigen polyclonal antibodies. Liver Int 26:116–124 doi:10.1111/j.1478-3231.2005.01201.x
Gomez-Lechon MJ, Castell JV, Donato MT (2007) Hepatocytes—the choice to investigate drug metabolism and toxicity in man: in vitro variability as a reflection of in vivo. Chem Biol Interact 168:30–50 doi:10.1016/j.cbi.2006.10.013
Soars MG, McGinnity DF, Grime K et al (2007) The pivotal role of hepatocytes in drug discovery. Chem Biol Interact 168:2–15 doi:10.1016/j.cbi.2006.11.002
Rodriguez-Antona C, Donato MT, Boobis A et al (2002) Cytochrome P450 expression in human hepatocytes and hepatoma cell lines: molecular mechanisms that determine lower expression in cultured cells. Xenobiotica 32:505–520 doi:10.1080/00498250210128675
Wilkening S, Stahl F, Bader A (2003) Comparison of primary human hepatocytes and hepatoma cell line Hepg2 with regard to their biotransformation properties. Drug Metab Dispos 31:1035–1042 doi:10.1124/dmd.31.8.1035
Henkens T, Papeleu P, Elaut G et al (2007) Trichostatin A, a critical factor in maintaining the functional differentiation of primary cultured rat hepatocytes. Toxicol Appl Pharmacol 218:64–71 doi:10.1016/j.taap.2006.10.012
Papeleu P, Loyer P, Vanhaecke T et al (2003) Trichostatin A induces differential cell cycle arrests but does not induce apoptosis in primary cultures of mitogen-stimulated rat hepatocytes. J Hepatol 39:374–382 doi:10.1016/S0168-8278(03)00288-5
Vanhaecke T, Henkens T, Kass GE et al (2004) Effect of the histone deacetylase inhibitor trichostatin A on spontaneous apoptosis in various types of adult rat hepatocyte cultures. Biochem Pharmacol 68:753–760 doi:10.1016/j.bcp.2004.05.022
Vinken M, Henkens T, Vanhaecke T et al (2006) Trichostatin a enhances gap junctional intercellular communication in primary cultures of adult rat hepatocytes. Toxicol Sci 91:484–492 doi:10.1093/toxsci/kfj152
Cabré J, Palomo AL (1984) New experimental strategies in amide synthesis using N,N-bis[2-oxo-3-oxazolidinyl]phosphorodiamidic chloride. Synthesis 5:413–417 doi:10.1055/s-1984-30857
Ho CY, Strobel E, Ralbovsky J et al (2005) Improved solution- and solid-phase preparation of hydroxamic acids from esters. J Org Chem 70:4873–4875 doi:10.1021/jo050036f
Jung M, Brosch G, Kolle D et al (1999) Amide analogues of trichostatin A as inhibitors of histone deacetylase and inducers of terminal cell differentiation. J Med Chem 42:4669–4679 doi:10.1021/jm991091h
Jung M, Hoffmann K, Brosch G et al (1997) Analogues of trichostatin A and trapoxin B as histone deacetylase inhibitors. Bioorg Med Chem Lett 7:1655–1658 doi:10.1016/S0960-894X(97)00284-9
Papeleu P, Vanhaecke T, Henkens T et al (2006) Isolation of rat hepatocytes. Methods Mol Biol 320:229–237
Menu E, Asosingh K, Van Riet I et al (2004) Myeloma cells (5TMM) and their interactions with the marrow microenvironment. Blood Cells Mol Dis 33:111–119 doi:10.1016/j.bcmd.2004.04.012
Elaut G, Torok G, Vinken M et al (2002) Major phase I biotransformation pathways of Trichostatin a in rat hepatocytes and in rat and human liver microsomes. Drug Metab Dispos 30:1320–1328 doi:10.1124/dmd.30.12.1320
Dai Y, Guo Y, Guo J et al (2003) Indole amide hydroxamic acids as potent inhibitors of histone deacetylases. Bioorg Med Chem Lett 13:1897–1901 doi:10.1016/S0960-894X(03)00301-9
Elaut G, Laus G, Alexandre E et al (2007) A metabolic screening study of trichostatin A (TSA) and TSA-like histone deacetylase inhibitors in rat and human primary hepatocyte cultures. J Pharmacol Exp Ther 321:400–408 doi:10.1124/jpet.106.116202
Blacklow RS (2007) Actuarially speaking: an overview of life expectancy. What can we anticipate. Am J Clin Nutr 86:1560S–1562S
Remiszewski SW, Sambucetti LC, Atadja P et al (2002) Inhibitors of human histone deacetylase: synthesis and enzyme and cellular activity of straight chain hydroxamates. J Med Chem 45:753–757 doi:10.1021/jm015568c
Elaut G, Rogiers V, Vanhaecke T (2007) The pharmaceutical potential of histone deacetylase inhibitors. Curr Pharm Des 13:2584–2620 doi:10.2174/138161207781663064
Wang DF, Helquist P, Wiech NL et al (2005) Toward selective histone deacetylase inhibitor design: homology modeling, docking studies, and molecular dynamics simulations of human class I histone deacetylases. J Med Chem 48:6936–6947 doi:10.1021/jm0505011
Acknowledgments
This study was financially supported by Vrije Universiteit Brussel (OZR-VUB), Belgium through the GOA project. Part of the project is also supported by the EU FP6 project, Liintop. Thanks go to Mrs. Pauwels M., Mr. Degreef B., Mr. Branson S. and Ms. Bastiaensen E. for their technical assistance.
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Joanna Fraczek and Sarah Deleu contributed equally to this article.
T. Vanhaecke is a postdoctoral research fellow of the Fund for Scientific Research Flanders (FWO-Vlaanderen, Belgium)
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Fraczek, J., Deleu, S., Lukaszuk, A. et al. Screening of amide analogues of Trichostatin A in cultures of primary rat hepatocytes: search for potent and safe HDAC inhibitors. Invest New Drugs 27, 338–346 (2009). https://doi.org/10.1007/s10637-008-9180-x
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DOI: https://doi.org/10.1007/s10637-008-9180-x