Selective, covalent modification of beta-tubulin residue Cys-239 by T138067, an antitumor agent with in vivo efficacy against multidrug-resistant tumors.

Microtubules are linear polymers of alpha- and beta-tubulin heterodimers and are the major constituents of mitotic spindles, which are essential for the separation of chromosomes during mitosis. Here we describe a synthetic compound, 2-fluoro-1-methoxy-4-pentafluorophenylsulfonamidobenzene (T138067), which covalently and selectively modifies the beta1, beta2, and beta4 isotypes of beta-tubulin at a conserved cysteine residue, thereby disrupting microtubule polymerization. Cells exposed to T138067 become altered in shape, indicating a collapse of the cytoskeleton, and show an increase in chromosomal ploidy. Subsequently, these cells undergo apoptosis. Furthermore, T138067 exhibits cytotoxicity against tumor cell lines that exhibit substantial resistance to vinblastine, paclitaxel, doxorubicin, and actinomycin D. T138067 is also equally efficacious in inhibiting the growth of sensitive and multidrug-resistant human tumor xenografts in athymic nude mice. These observations suggest that T138067 may be clinically useful for the treatment of multidrug-resistant tumors.

[1]  R. Luduena,et al.  Tubulin sulfhydryl groups as probes and targets for antimitotic and antimicrotubule agents. , 1991, Pharmacology & therapeutics.

[2]  Y. Rustum,et al.  Mechanisms of resistance to fluoropyrimidines. , 1992, Seminars in oncology.

[3]  G. Bloom.  Motor proteins for cytoplasmic microtubules , 1992, Current Biology.

[4]  M. Gottesman,et al.  2,4-Dichlorobenzyl thiocyanate, an antimitotic agent that alters microtubule morphology. , 1986, Proceedings of the National Academy of Sciences of the United States of America.

[5]  D. Kingston,et al.  Characterization of the Taxol Binding Site on the Microtubule , 1995, The Journal of Biological Chemistry.

[6]  K. Sullivan,et al.  Molecular biology and genetics of tubulin. , 1985, Annual review of biochemistry.

[7]  D. Sackett.  Podophyllotoxin, steganacin and combretastatin: natural products that bind at the colchicine site of tubulin. , 1993, Pharmacology & therapeutics.

[8]  T. Mitchison.  Microtubule dynamics and kinetochore function in mitosis. , 1988, Annual review of cell biology.

[9]  U. Eppenberger,et al.  The dual role of mutant p53 protein in chemosensitivity of human cancers. , 1996, Anticancer research.

[10]  J. Nath,et al.  Effects of caffeine and other methylxanthines on the development and metabolism of sea urchin eggs. Involvement of NADP and glutathione , 1976, The Journal of cell biology.

[11]  R. Tjian,et al.  Coactivator and Promoter-Selective Properties of RNA Polymerase I TAFs , 1995, Science.

[12]  R. Donehower,et al.  The clinical pharmacology and use of antimicrotubule agents in cancer chemotherapeutics. , 1991, Pharmacology & therapeutics.

[13]  Kenneth H. Downing,et al.  Structure of the αβ tubulin dimer by electron crystallography , 1998, Nature.

[14]  A. Hyman,et al.  Coupling cell division and cell death to microtubule dynamics. , 1997, Current opinion in cell biology.

[15]  E. Rowinsky.  The development and clinical utility of the taxane class of antimicrotubule chemotherapy agents. , 1997, Annual review of medicine.

[16]  H. Sakai,et al.  Role of tubulin-SH groups in polymerization to microtubules. Functional-SH groups in tubulin for polymerization. , 1974, Journal of biochemistry.

[17]  A. Brossi,et al.  Identification of Cysteine 354 of β-Tubulin as Part of the Binding Site for the A Ring of Colchicine (*) , 1996, The Journal of Biological Chemistry.

[18]  I. Pastan,et al.  P-glycoprotein and multidrug resistance. , 1996, Current opinion in genetics & development.

[19]  E. Hamel.  Antimitotic natural products and their interactions with tubulin , 1996, Medicinal research reviews.

[20]  M. Jordan,et al.  Microtubule dynamics: taking aim at a moving target. , 1995, Chemistry & biology.

[21]  M. Steiner,et al.  Sulfhydryls of platelet tubulin: their role in polymerization and colchicine binding. , 1978, Biochemistry.

[22]  K. Sullivan.  Structure and utilization of tubulin isotypes. , 1988, Annual review of cell biology.

[23]  B. Bhattacharyya,et al.  Anion-induced increases in the rate of colchicine binding to tubulin. , 1976, Biochemistry.

[24]  M. Jordan,et al.  Microtubules and actin filaments: dynamic targets for cancer chemotherapy. , 1998, Current opinion in cell biology.

[25]  A. Bershadsky,et al.  Microtubule dynamics: mechanism, regulation, and function. , 1991, Annual review of cell biology.

[26]  S. Cole,et al.  Function, evolution and structure of multidrug resistance protein (MRP). , 1997, Seminars in cancer biology.

[27]  T. Liu,et al.  Identification of the cysteine residue of beta-tubulin alkylated by the antimitotic agent 2,4-dichlorobenzyl thiocyanate, facilitated by separation of the protein subunits of tubulin by hydrophobic column chromatography. , 1989, Biochemistry.

[28]  Amanda G Paulovich,et al.  When Checkpoints Fail , 1997, Cell.

[29]  I. Pastan,et al.  Biochemistry of multidrug resistance mediated by the multidrug transporter. , 1993, Annual review of biochemistry.

[30]  R. Himes,et al.  Inhibition of tubulin self-assembly in vitro by fluorodinitrobenzene. , 1981, Biochimica et biophysica acta.

[31]  R. Luduena,et al.  Preparation of a monoclonal antibody specific for the class IV isotype of beta-tubulin. Purification and assembly of alpha beta II, alpha beta III, and alpha beta IV tubulin dimers from bovine brain. , 1992, The Journal of biological chemistry.

[32]  J. Wolff,et al.  Localization of the Vinblastine-binding Site on β-Tubulin* , 1996, The Journal of Biological Chemistry.

[33]  R. Luduena.  Are tubulin isotypes functionally significant. , 1993, Molecular biology of the cell.

[34]  T. Chou,et al.  Synergism and antagonism in chemotherapy , 1991 .

[35]  C. Ramel.  Geetic effects of organic mercury compounds. I. Cytological investigations on Allium roots. , 2009, Hereditas.