Screening for Specific Inhibitors of Src Family Kinases

筛选 Src 家族激酶的特异性抑制剂

基本信息

批准号：
63870106
负责人：
IBA Hideo
金额：
$ 5.89万
依托单位：
Institute of Medical Science, University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for Developmental Scientific Research (B).
财政年份：
1988
资助国家：
日本
起止时间：
1988 至 1990
项目状态：
已结题

项目摘要

1. In 1987, we reported that Staurosporine can inhibit several species of protein kinases at very low concentrations (nMorder), we started this project by synthesizing more than 50 derivatives of Staurosporine. Using v-src kinase as well as abl kinase synthesized in E. coli, as the tyrosine kinases, we had screened these derivatives but non of them showed specific inhibition at a low concentration (about 100nM).2. Genistein and quercetin that were previously reported as the inhibitors specific to tyrosine kinases were tested in our system. IR^<50> values we determined were, however, were much higher than reported previously. We have also shown that these two reagents have DNA cleaving activity that is DNA topoisomerase dependent. These results indicate that these regents have clear limitation to be used as specific kinase inhibitor in vivo.3. In this project, we isolated a specific inhibitor of protein kinase C from microbiral extract, which was named as UCN1028. We further purified and found that the specific compound, Calphostin C, in UCN1028 is responsible for the specific inhibition. Calphostin C showed very strong antitumor activity when injected into tumor bearing mouse.4. We recently isolated new fos related gene, fra-2. This gene was found to be one member of immediate early genes and to have transforming activity. Induction of the mRNA of this gene was shown to be mediated by several kinds of tyrosine kinase or serine kinase and its gene product, Fra-2 was further shown to be phosphorylated by serum inducible protein kinase activity. This Fra-2 expression as well as modification is expected to be sensitive and effective indicators for the screening of new kinase inhibitors in vivo.

1。在1987年，我们报告说，星形孢菌素可以以非常低浓度（Nmorder）抑制几种蛋白质激酶，我们通过合成50多个星形孢菌素的衍生物来启动该项目。我们使用大肠杆菌中的V-SRC激酶以及ABL激酶合成的ABL激酶，作为酪氨酸激酶，我们筛选了这些衍生物，但其中未在低浓度（约100nm）时显示出特定的抑制作用。2。先前报道为酪氨酸激酶特有的抑制剂的染料黄酮和槲皮素在我们的系统中进行了测试。但是，我们确定的IR^<50>值比以前报道的要高得多。我们还表明，这两种试剂具有DNA裂解活性，即DNA拓扑异构酶依赖性。这些结果表明，这些摄政量在体内具有明显的限制，可以用作特定的激酶抑制剂3。在这个项目中，我们从微病毒提取物中分离出蛋白激酶C的特异性抑制剂，该蛋白激酶C被称为UCN1028。我们进一步纯化，发现UCN1028中的特定化合物Calphostin C负责特定的抑制作用。当注射到肿瘤轴承小鼠中时，钙粘蛋白C表现出非常强的抗肿瘤活性。4。我们最近分离了新的相关基因FRA-2。该基因被发现是立即基因的成员，并且具有转化活性。该基因的mRNA诱导被证明是由几种酪氨酸激酶或丝氨酸激酶及其基因产物介导的，FRA-2进一步证明是通过血清诱导蛋白激酶活性磷酸化的。这种FRA-2表达以及修饰有望是敏感有效的指标，用于筛选新的激酶抑制剂体内。