The overall objectives of this research proposal are the further development of a multidisciplinary program for studying the biochemical mechanism of action of, and the molecular basis of resistance to, antitumor drugs. The ultimate goal is to use the knowledge acquired in these studies to: 1) develop taxol analogs that will maintain or improve the therapeutic activity of the drug and be amenable to production in concentrations required for clinical studies and 2) design therapeutic protocols to avoid and circumvent drug resistance. The specific objectives of this research proposal are to: 1) understand the molecular mechanisms by which cells become resistant to taxol. Taxol-resistant cells isolated in the laboratory will be analyzed for the presence of P-glycoprotein and altered forms of tubulin. Cell lines grown in the presence of taxol and cis-platinum will be isolated and analyzed for drug resistance. 2) prepare antibodies to taxol that can be used for a) visualizing the drug in normal and tumor cells by immunofluorescence and on the microtubule by immunoelectron microscopy utilizing gold spheres, b) developing an radioimmune assay (RIA) for the quantitation of taxol in body fluids and tissues, and in crude extracts of plants, and c) domain mapping of the taxol binding site in the microtubule. 3) determine the drug binding site(s) on the microtubule by using photoaffinity labeled radioactive taxol analogs that will covalently bind to the microtubule. Isolation and analysis of a peptide(s) containing the radiolabeled taxol analog should provide the amino acid sequence with which taxol interacts. This information, taken together with the known structural features of taxol that are required for binding, plus what is known about the tertiary structure of tubulin, should produce, at least a partial definition of the binding site for taxol on the microtubule. 4) further define the structure-activity profile of taxol with specific emphasis on the A-ring side chain. Such information will provide insight into the chemical features of taxol that are important in its interaction with microtubules and allow the design of new therapeutic analogues.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Unknown (R35)
Project #
5R35CA039821-09
Application #
3479106
Study Section
Special Emphasis Panel (SRC (88))
Project Start
1985-08-01
Project End
1999-07-31
Budget Start
1993-08-01
Budget End
1994-07-31
Support Year
9
Fiscal Year
1993
Total Cost
Indirect Cost
Name
Albert Einstein College of Medicine
Department
Type
Schools of Medicine
DUNS #
009095365
City
Bronx
State
NY
Country
United States
Zip Code
10461
Mani, S; McDaid, H M; Grossman, A et al. (2007) Peripheral blood mononuclear and tumor cell pharmacodynamics of the novel epothilone B analogue, ixabepilone. Ann Oncol 18:190-5
Ikui, Amy E; Yang, Chia-Ping Huang; Matsumoto, Tomohiro et al. (2005) Low concentrations of taxol cause mitotic delay followed by premature dissociation of p55CDC from Mad2 and BubR1 and abrogation of the spindle checkpoint, leading to aneuploidy. Cell Cycle 4:1385-8
Mani, Sridhar; Huang, Haiyan; Sundarababu, Sumathy et al. (2005) Activation of the steroid and xenobiotic receptor (human pregnane X receptor) by nontaxane microtubule-stabilizing agents. Clin Cancer Res 11:6359-69
Klein, Laura E; Freeze, B Scott; Smith 3rd, Amos B et al. (2005) The microtubule stabilizing agent discodermolide is a potent inducer of accelerated cell senescence. Cell Cycle 4:501-7
Geney, Raphael; Sun, Liang; Pera, Paula et al. (2005) Use of the tubulin bound paclitaxel conformation for structure-based rational drug design. Chem Biol 12:339-48
Mani, Sridhar; McDaid, Hayley; Hamilton, Anne et al. (2004) Phase I clinical and pharmacokinetic study of BMS-247550, a novel derivative of epothilone B, in solid tumors. Clin Cancer Res 10:1289-98
Chen, Jie-Guang; Yang, Chia-Ping Huang; Cammer, Michael et al. (2003) Gene expression and mitotic exit induced by microtubule-stabilizing drugs. Cancer Res 63:7891-9
Verdier-Pinard, Pascal; Wang, Fang; Burd, Berta et al. (2003) Direct analysis of tubulin expression in cancer cell lines by electrospray ionization mass spectrometry. Biochemistry 42:12019-27
Verdier-Pinard, Pascal; Wang, Fang; Martello, Laura et al. (2003) Analysis of tubulin isotypes and mutations from taxol-resistant cells by combined isoelectrofocusing and mass spectrometry. Biochemistry 42:5349-57
Martello, Laura A; Verdier-Pinard, Pascal; Shen, Heng-Jia et al. (2003) Elevated levels of microtubule destabilizing factors in a Taxol-resistant/dependent A549 cell line with an alpha-tubulin mutation. Cancer Res 63:1207-13

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