TAS-301, a New Synthetic Inhibitor of Neointimal Thickening after Balloon Injury, Inhibits Calcium-Dependent Signal Transduction and Cytoskeletal Reorganization

We previously demonstrated that a recently synthesized drug, TAS-301 [3-bis(4-methoxyphenyl)methylene-2-indolinone], inhibited neointimal thickening after single-balloon injury to the rat common carotid artery by inhibiting both the migration and proliferation processes of vascular smooth muscle cells (VSMCs). The purpose of this current study was to elucidate the possible mechanism of action for its inhibition of the migration process of VSMCs. We also determined the efficacy of TAS-301 on second neointimal formation 14 days after a double-balloon injury to the rat common carotid artery. Neointimal thickening, 14 days after second balloon injury, was reduced by the oral administration of TAS-301 in a dose-dependent manner. In in vitro assays using rat VSMCs, Western blot analysis showed that TAS-301 inhibited platelet-derived growth factor (PDGF)-induced tyrosine phosphorylation of both focal adhesion kinase and paxillin. Tyrosine phosphorylation of these proteins depended on the increment of intracellular calcium concentration ([Ca2+]i). The PDGF-induced elevation of [Ca2+]i and activation of Ca2+/calmodulin-dependent protein kinase II (CaM kinase II) were also inhibited by TAS-301. Additionally, TAS-301 at 10 micromol/l reduced the extent of F-actin stress fiber depolymerization observed in response to PDGF. These results indicate that TAS-301 reduced the intimal thickening after denudation to a pre-existing lesion to the same extent as it reduced that after a single-balloon injury to the normal artery. Furthermore, the results of our in vitro experiments suggest that antimigratory mechanisms of TAS-301 that contribute to preventing the intimal thickening might be mediated by drug inhibition of Ca2+ -dependent signal molecules and the following cytoskeletal depolymerization.