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，有必要为抗癌药物的发现确定新的分子靶点。其中包括直接调节固有的细胞凋亡途径的分子。我们最近发现，大的GTP酶Dynamin-2(Dyn2)通过P53信号触发细胞凋亡，特别是在快速分裂的细胞中。仅5倍的内源性野生型dyn2过表达就会激活p53依赖的细胞凋亡，这完全依赖于GTP的结合。动力蛋白固有的间隙活性存在缺陷的突变甚至更有效。因此，抑制Dynamin的GTPase活性或其固有间隙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，我们开发了一种简单而可靠的动力蛋白GTP酶活性高通量分析方法，并与领先的化学家巴里·夏普莱斯建立了合作关系，他开创了一种基于叠氮的新方法，该方法有助于生成相关化合物的复杂文库，允许对已识别的先导进行快速模块化SAR，最重要的是，最重要的是，唯一允许在目标分子表面为其选择组装的功能部分，以生成定制的、高亲和力、高特异性的抑制剂。我们提出了以下具体目标，使我们能够应用点击化学来确定针对调节GTP酶Dynamin的潜在抗癌药物。 1.鉴定、鉴定和优化抑制Dynamin基础GTP酶活性的高亲和力、高特异性化合物。 2.鉴定、鉴定和优化选择性抑制Dynamin组装刺激的GTP酶活性的高亲和力、高特异性化合物。 3.应用点击化学原位筛选对Dynamin本身具有极高亲和力、极高特异性的抑制剂。

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.