FK228 (depsipeptide) as a natural prodrug that inhibits class I histone deacetylases.

FK228 is a histone deacetylase (HDAC) inhibitor, the molecular mechanism of inhibition of which has been unknown. Here we show that reduction of an intramolecular disulfide bond of FK228 greatly enhanced its inhibitory activity and that the disulfide bond was rapidly reduced in cells by cellular reducing activity involving glutathione. Computer modeling suggests that one of the sulfhydryl groups of the reduced form of FK228 (redFK) interacts with the active-site zinc, preventing the access of the substrate. HDAC1 and HDAC2 were more strongly inhibited by redFK than HDAC4 and HDAC6. redFK was less active than FK228 in inhibiting in vivo HDAC activity, due to rapid inactivation in medium and serum. Thus, FK228 serves as a stable prodrug to inhibit class I enzymes and is activated by reduction after uptake into the cells. The glutathione-mediated activation also implicates its clinical usefulness for counteracting glutathione-mediated drug resistance in chemotherapy.

[1]  R. Huber,et al.  X-ray structures of human neutrophil collagenase complexed with peptide hydroxamate and peptide thiol inhibitors. Implications for substrate binding and rational drug design. , 1995, European journal of biochemistry.

[2]  Structure of the lysosomal sphingolipid activator protein 1 by homology with influenza virus neuraminidase. , 1988, Biochemical and biophysical research communications.

[3]  J. Essigmann,et al.  Mechanisms of resistance to cisplatin. , 2001, Mutation research.

[4]  B. Matthews,et al.  Structure of a mercaptan-thermolysin complex illustrates mode of inhibition of zinc proteases by substrate-analogue mercaptans. , 1982, Biochemistry.

[5]  T. Tsuruo,et al.  Cyclic hydroxamic-acid-containing peptide 31, a potent synthetic histone deacetylase inhibitor with antitumor activity. , 2001, Cancer research.

[6]  S. Doglia,et al.  Regulation of cellular glutathione modulates nuclear accumulation of daunorubicin in human MCF7 cells overexpressing multidrug resistance associated protein. , 2000, European journal of cancer.

[7]  S. Inoue,et al.  Role of the histone deacetylase complex in acute promyelocytic leukaemia , 1998, Nature.

[8]  H. Kwon,et al.  Histone deacetylase inhibitor FK228 inhibits tumor angiogenesis , 2002, International journal of cancer.

[9]  J. Powers,et al.  Design of potent reversible inhibitors for thermolysin. Peptides containing zinc coordinating ligands and their use in affinity chromatography. , 1979, Biochemistry.

[10]  S. Meng,et al.  p21WAF1 is required for butyrate-mediated growth inhibition of human colon cancer cells , 1998 .

[11]  C. Tribioli,et al.  Distinct interactions of PML-RARα and PLZF-RARα with co-repressors determine differential responses to RA in APL , 1998, Nature Genetics.

[12]  A. Wolffe,et al.  Activators and repressors: making use of chromatin to regulate transcription , 1997, Genes to cells : devoted to molecular & cellular mechanisms.

[13]  N. Mutoh,et al.  Isolation of mutants of Schizosaccharomyces pombe unable to synthesize cadystin, small cadmium-binding peptides. , 1988, Biochemical and biophysical research communications.

[14]  S. Minucci,et al.  Fusion proteins of the retinoic acid receptor-α recruit histone deacetylase in promyelocytic leukaemia , 1998, Nature.

[15]  S. Schreiber,et al.  A Mammalian Histone Deacetylase Related to the Yeast Transcriptional Regulator Rpd3p , 1996, Science.

[16]  C. Glass,et al.  Co-activators and co-repressors in the integration of transcriptional responses. , 1998, Current opinion in cell biology.

[17]  Bing-he Xu,et al.  Effect of buthionine sulfoximine and ethacrynic acid on cytotoxic activity of mitomycin C analogues BMY 25282 and BMY 25067. , 1992, Cancer research.

[18]  M. Yoshida,et al.  FR901228, a potent antitumor antibiotic, is a novel histone deacetylase inhibitor. , 1998, Experimental cell research.

[19]  I. Kawamura,et al.  FR901228, a Novel Antitumor Bicyclic Depsipeptide Produced by Chromobacterium violaceum No. 968. Part 3. Antitumor Activities on Experimental Tumors in Mice. , 1994 .

[20]  G. Meersma,et al.  Relationship of cellular glutathione to the cytotoxicity and resistance of seven platinum compounds. , 1992, Cancer research.

[21]  E. Sausville,et al.  P21-dependent G 1 arrest with downregulation of cyclin D1 and upregulation of cyclin E by the histone deacetylase inhibitor FR901228 , 2000, British Journal of Cancer.

[22]  D. Faller,et al.  Butyrate-induced G1 arrest results from p21-independent disruption of retinoblastoma protein-mediated signals. , 1998, Cell growth & differentiation : the molecular biology journal of the American Association for Cancer Research.

[23]  S. Horinouchi,et al.  Trichostatin A and trapoxin: Novel chemical probes for the role of histone acetylation in chromatin structure and function , 1995, BioEssays : news and reviews in molecular, cellular and developmental biology.

[24]  T. Fojo,et al.  Low concentrations of the histone deacetylase inhibitor, depsipeptide (FR901228), increase expression of the Na(+)/I(-) symporter and iodine accumulation in poorly differentiated thyroid carcinoma cells. , 2001, The Journal of clinical endocrinology and metabolism.

[25]  B. Matthews,et al.  Binding of hydroxamic acid inhibitors to crystalline thermolysin suggests a pentacoordinate zinc intermediate in catalysis. , 1982, Biochemistry.

[26]  L. Neckers,et al.  Modulation of p53, ErbB1, ErbB2, and Raf-1 expression in lung cancer cells by depsipeptide FR901228. , 2002, Journal of the National Cancer Institute.

[27]  R. Huber,et al.  The X‐ray crystal structure of the catalytic domain of human neutrophil collagenase inhibited by a substrate analogue reveals the essentials for catalysis and specificity. , 1994, The EMBO journal.

[28]  M. Grunstein,et al.  HDA1 and RPD3 are members of distinct yeast histone deacetylase complexes that regulate silencing and transcription. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[29]  S. Meng,et al.  WAF 1 is required for butyrate-mediated growth inhibition of human colon cancer cells , 1998 .

[30]  J. Powers,et al.  Inhibition of thermolysin and carboxypeptidase A by phosphoramidates. , 1979, Biochemistry.

[31]  M. Okuhara,et al.  FR901228, a novel antitumor bicyclic depsipeptide produced by Chromobacterium violaceum No. 968. I. Taxonomy, fermentation, isolation, physico-chemical and biological properties, and antitumor activity. , 1994, The Journal of antibiotics.

[32]  M. Okuhara,et al.  Action of FR901228, a novel antitumor bicyclic depsipeptide produced by Chromobacterium violaceum no. 968, on Ha-ras transformed NIH3T3 cells. , 1994, Bioscience, biotechnology, and biochemistry.

[33]  W. Wilson,et al.  Inhibitor of histone deacetylation, depsipeptide (FR901228), in the treatment of peripheral and cutaneous T-cell lymphoma: a case report. , 2001, Blood.

[34]  B Rubin,et al.  Design of specific inhibitors of angiotensin-converting enzyme: new class of orally active antihypertensive agents. , 1977, Science.

[35]  T. Ishikawa,et al.  The Human Multidrug Resistance-Associated Protein (MRP) Gene Family: From Biological Function to Drug Molecular Design , 2000, Clinical chemistry and laboratory medicine.

[36]  R. Huber,et al.  X‐ray Structures of Human Neutrophil Collagenase Complexed with Peptide Hydroxamate and Peptide Thiol Inhibitors , 1995 .

[37]  L. Guarente,et al.  Transcriptional silencing and longevity protein Sir2 is an NAD-dependent histone deacetylase , 2000, Nature.

[38]  A. Verdel,et al.  Functional significance of histone deacetylase diversity. , 2001, Current opinion in genetics & development.

[39]  M. Yoshida,et al.  Potent histone deacetylase inhibitors built from trichostatin A and cyclic tetrapeptide antibiotics including trapoxin. , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[40]  M. Yoshida,et al.  Trapoxin, an antitumor cyclic tetrapeptide, is an irreversible inhibitor of mammalian histone deacetylase. , 1993, The Journal of biological chemistry.

[41]  S. Horinouchi,et al.  Oxamflatin is a novel antitumor compound that inhibits mammalian histone deacetylase , 1999, Oncogene.

[42]  S. Waxman,et al.  Malignant cells can be sensitized to undergo growth inhibition and apoptosis by arsenic trioxide through modulation of the glutathione redox system. , 1999, Blood.

[43]  P. Marks,et al.  Structures of a histone deacetylase homologue bound to the TSA and SAHA inhibitors , 1999, Nature.

[44]  N. Borkakoti,et al.  Structure of the catalytic domain of human fibroblast collagenase complexed with an inhibitor , 1994, Nature Structural Biology.

[45]  S. Meng,et al.  p21(WAF1) is required for butyrate-mediated growth inhibition of human colon cancer cells. , 1998, Proceedings of the National Academy of Sciences of the United States of America.