Ali Nermin, Yoshizumi Masanori, Fujita Yoshiko, Izawa Yuki, Kanematsu Yasuhisa, Ishizawa Keisuke, Tsuchiya Koichiro, Yano Seiji, Sone Saburo, Tamaki Toshiaki
Department of Pharmacology, The University of Tokushima Graduate School Institute of Health Biosciences, Japan.
J Pharmacol Sci. 2005 Jun;98(2):130-41. doi: 10.1254/jphs.fp0040850. Epub 2005 Jun 4.
Vascular endothelial growth factor (VEGF) was reported to be a potent proangiogenic factor that plays a pivotal role in both physiological and pathological angiogenesis. M475271, 4-quinazolinamine, N-(2-chloro-5-methoxyphenyl)-6-methoxy-7-[(1-methyl-4-piperidinyl) methoxy]-(9Cl), is a new anilinoquinazoline derivative that showed selective inhibition of Src kinase activity and tumor growth in vivo. Here, we examined the effect of M475271 on VEGF-induced human umbilical vein endothelial cell (HUVEC) proliferation and migration and their intracellular mechanisms. Our findings showed that M475271 pretreatment resulted in a significant inhibition of VEGF-induced HUVEC proliferation, [(3)H]thymidine incorporation, and migration. M475271 inhibited VEGF-induced Flk-1 and Src phosphorylation and their association. Confocal laser microscopic examination confirmed the inhibitory effect of M475271 on VEGF-induced Flk-1/Src association. M475271 inhibited VEGF-induced extracellular signal-regulated kinase1/2 (ERK1/2) and p38 but not Akt activation in a concentration-dependent manner. M475271, PI3-K inhibitor, and p38 inhibitor inhibited VEGF-induced HUVEC proliferation and migration. However, a MEK1/2 inhibitor inhibited VEGF-induced proliferation but not migration. These findings suggest that M475271 attenuates VEGF-induced HUVEC proliferation and migration through the inhibition of signaling pathways involving Src, ERK1/2, and/or p38. Taken together, these data indicate that M475271 may be a useful candidate for inhibition of endothelial cell proliferation and migration relevant to angiogenesis.
血管内皮生长因子（VEGF）据报道是一种强效的促血管生成因子，在生理和病理血管生成中均起关键作用。M475271，即4-喹唑啉胺，N-（2-氯-5-甲氧基苯基）-6-甲氧基-7-[(1-甲基-4-哌啶基)甲氧基]-(9Cl)，是一种新型苯胺基喹唑啉衍生物，在体内表现出对Src激酶活性和肿瘤生长的选择性抑制作用。在此，我们研究了M475271对VEGF诱导的人脐静脉内皮细胞（HUVEC）增殖和迁移及其细胞内机制的影响。我们的研究结果表明，M475271预处理可显著抑制VEGF诱导的HUVEC增殖、[³H]胸苷掺入及迁移。M475271抑制VEGF诱导的Flk-1和Src磷酸化及其相互作用。共聚焦激光显微镜检查证实了M475271对VEGF诱导的Flk-1/Src相互作用的抑制作用。M475271以浓度依赖性方式抑制VEGF诱导的细胞外信号调节激酶1/2（ERK1/2）和p38激活，但不抑制Akt激活。M475271、PI3-K抑制剂和p38抑制剂均抑制VEGF诱导的HUVEC增殖和迁移。然而，MEK1/2抑制剂抑制VEGF诱导的增殖但不抑制迁移。这些研究结果表明，M475271通过抑制涉及Src、ERK1/2和/或p38的信号通路来减弱VEGF诱导的HUVEC增殖和迁移。综上所述，这些数据表明M475271可能是抑制与血管生成相关的内皮细胞增殖和迁移的有用候选药物。
