Interaction of 4-Arylcoumarin Analogues of Combretastatins with Microtubule Network of HBL100 Cells and Binding to Tubulin
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- 作者：Catherine Rappl ; Pascale Barbier ; Vé ; ronique Bourgarel-Rey ; Catherine Gré ; goire ; Robert Gilli ; Manon Carre ; Sé ; bastien Combes ; Jean-Pierre Finet ; Vincent Peyrot
- 刊名：Biochemistry
- 出版年：2006
- 出版时间：August 1, 2006
- 年：2006
- 卷：45
- 期：30
- 页码：9210 - 9218
- 全文大小：327K
- 年卷期：v.45,no.30(August 1, 2006)
- ISSN：1520-4995
文摘
The synthesis of different 4-arylcoumarin analogues of combretastatin A-4 led to theidentification of two new compounds (1 and 2) with potent cytotoxic activity on a CEM leukemia cellline and a third one completely inactive (compound 3). It was suggested that the cytotoxicity of compounds1 and 2 may be related to their interaction with microtubules and tubulin, since these compounds inhibitmicrotubule formation from purified tubulin in vitro [Bailly et al. (2003) J. Med. Chem. 46 (25), 5437-5444]. In the present study, tubulin was identified as the main target of these molecules. We studiedstructure-activity relationships of these compounds using biological experiments specific for tubulinbinding. The modification of cell cycle progression induced by compounds 1 and 2 was characterized byan apoptotic induction on human breast cells (HBL100). In addition, these two molecules disturbed cellsurvival by depolymerizing the microtubule network, leading to a mitotic block. We then determined thethermodynamic parameters of their interaction with purified tubulin by fluorescence spectroscopy andisothermal microcalorimetry. These results, together with a superimposition of the molecule on colchicinein the X-ray-determined three-dimensional structure model of tubulin-colchicine complex, allowed us toidentify the pharmacophore of the combretastatin A-4 analogues responsible for their biological activity.