Inhibitors of Dynamin as Novel Anti-Cancer Therapeutics

描述（由申请人提供）：正如本RFA所认识到的，需要确定用于癌症药物发现的新型分子靶标。其中将是直接调节内在凋亡途径的分子。我们最近发现，大的GT3发动蛋白-2（dyn 2）通过p53信号触发凋亡，特别是在快速分裂的细胞中。低至5倍的内源性野生型dyn 2过表达将激活p53依赖性细胞凋亡，这是绝对依赖于GTP结合。发动蛋白内在GAP活性缺陷的突变甚至更有效。因此，抑制发动蛋白的GTP酶活性或其内在GAP的化合物可能是有效的促凋亡抗癌药物。为了鉴定这些化合物，我们开发了一种用于发动蛋白GT3活性的简单而稳健的高通量测定法，并与领先的化学家巴里·夏普勒斯建立了合作关系，他开创了一种基于叠氮基的“点击化学”新方法，该方法有助于生成相关化合物的复杂文库，并且最重要且独特地允许在靶分子表面上选择功能部分的组装以产生定制的、高亲和力、高特异性的抑制剂。我们提出了以下具体目标，这将使我们能够应用点击化学来识别针对调节性GTdR，发动蛋白的潜在抗癌药物。 1.鉴定、表征和优化抑制发动蛋白基础GT3活性的高亲和力、高特异性化合物。 2.鉴定、表征和优化高亲和力、高特异性的化合物，其选择性抑制发动蛋白的组装刺激的GT3活性。 3.应用点击化学原位选择对发动蛋白本身具有很高亲和力、很高特异性的抑制剂。

DESCRIPTION (provided by applicant): As realized by this RFA, there is a need to identify novel molecular targets for cancer drug discovery. Among these will be molecules that directly regulate the intrinsic apoptotic pathway. We recently discovered that the large GTPase dynamin-2 (dyn2) signals through p53 to trigger apoptosis, specifically in rapidly dividing cells. As little as five-fold overexpression of endogenous wild-type dyn2 will activate p53-dependent apoptosis and this is absolutely dependent on GTP binding. A mutation defective in dynamin's intrinsic GAP activity is even more potent. Thus, compounds that inhibit dynamin's GTPase activity or its intrinsic GAP may be effective proapoptotic, anti-cancer drugs. To identify these compounds we have developed a simple and robust high-throughput assay for dynamin GTPase activity and established a collaboration with Barry Sharpless, a leading chemist who has pioneered a new azido-based method of "click chemistry" that facilitates the generation of complex libraries of related compounds, allows for rapid modular SAR of identified leads and most importantly and uniquely allows the assembly of functional moieties to be selected for on the surface of the target molecule to generate customized, high-affinity, high-specificity inhibitors. We propose the following Specific Aims that will allow us to apply click chemistry to identify potential anti-cancer drugs targeted against the regulatory GTPase, dynamin. 1. To identify, characterize and optimize high affinity, high specificity compounds that inhibit dynamin's basal GTPase activity. 2. To identify, characterize and optimize high affinity, high specificity compounds that selectively inhibit dynamin's assembly-stimulated GTPase activity. 3. To apply click chemistry for in situ selection of very high affinity, very high specificity inhibitors on dynamin, itself.