A collaborative structural hub to promote Cancer Center science and drug design

摘要 该项目的目标是为SBPMDI癌症中心的推广提供一个协作性的结构性“枢纽” 科学，药物发现，基于结构的药物设计和抑制机制。要了解 已发现的探针/先导化合物的机理和/或为了增强其效力， 需要带有目标蛋白的探针。这种类型的工作通常需要故障排除和优化， 由于筛选得到的化合物往往与未知亲和力结合，存在溶解性问题， 使共结晶复杂化和/或在未知的变构位置结合。我已经在六个项目上取得了成功 与SBPMDI癌症中心的首席调查人员合作。这些项目中有四个 NCI资金(RXRα配体结合域、HIF2HIF2PASSB域、PLEKHA7PH域、Fas TE域)和 两个是目前正在审查的NCI申请的一部分(TA03激酶域和ALDOA)。RXRα工作正常 揭示了非常相似的先导化合物之间不同的抑制机制。HIF2αPSB工作 展示了严格抑制的结构基础。PLEKHA7 PH和TAO 3激活区是新发现的 具有结合配体的晶体结构；在第一种情况下，揭示了底物特异性的结构基础和 第二种情况，指出了一种增强抑制剂特异性的途径。ALDOA项目揭示了 新型变构共价抑制剂的结构基础，以及可能的S-亚硝化位点 确定亚细胞定位。所有六个目标都是新的(PLEKHA7、TA03和ALDOA)，或已建立 (RXRα，HIF2α和Fas TE)癌症靶点，我计划继续在它们的进一步开发上合作 从击球到领先。我还在列表中添加了CNK1的PH结构域，它是一种与KRAS相关的癌症靶点，具有独特的 属性。根据我不同的经验，我开发了一种成本效益高的方法 成功的可能性，采用生物物理分析、核磁共振、高通量结晶和精细聚焦 同步辐射。我还鼓励和培训博士后、学生、教职员工科学家和 希望参与结构研究的技术人员。NCI R50奖将让我有一个坚定的财务 在此基础上扩展当前研究并启动新的合作，以包括具有新的 发现了目标或探测器，但目前缺乏资金来支持我的协作工作。 需要了解新药候选药物的结构和机制方面才能移动受攻击的化合物 用于引导和临床测试漏斗，并帮助主要研究人员为他们的研究获得资金， 从而加快了基础科学发现从“长凳到床边”的移动。

ABSTRACT The goal of this project is to provide a collaborative structural "hub" to promote SBPMDI Cancer Center science, drug discovery, structure-based drug design and inhibitory mechanism. To understand the mechanism of a discovered probe/lead compound and/or to enhance its potency, a co-crystal structure of the probe with a target protein is required. This type of work typically requires troubleshooting and optimization, because compounds obtained by screening often bind with unknown affinity, have solubility issues which complicate co-crystallization and/or bind at unknown allosteric sites. I have already succeeded on six project fronts in collaboration with Principal Investigators at the SBPMDI Cancer Center. Four of these projects have NCI funding (RXRα ligand binding domain, Hif2α PASB domain, PLEKHA7 PH domain, FAS TE domain) and two are part of NCI applications currently under review (TAO3 kinase domain and ALDOA). The RXRα work revealed distinct inhibitory mechanisms between very similar lead compounds. The Hif2α PASB work demonstrated the structural basis of tight inhbition. The PLEKHA7 PH and TAO3 kinase domains are novel crystal structures with bound ligand; in the first case, revealing a structural basis for substrate specificity and in the second case, pointing to a pathway to enhance inhibitor specificity. The ALDOA project has revealed the structural basis of novel allosteric covalent inhibitors, as well as potential S-nitrosylation sites that may determine subcellular localization. All six targets are novel (PLEKHA7, TAO3 and ALDOA), or established (RXRα, Hif2α and FAS TE) cancer targets, and I plan to continue collaborations on their further development from hit-to-lead. I have also added to the list the PH domain of CNK1, a KRas-linked cancer target with unique properties. Based on my varied experiences, I have developed a cost efficient methodology with a high likelihood of success, employing biophysical analysis, NMR, high-throughput crystallization and fine-focus synchrotron radiation. I also encourage and provide training to postdocs, students, staff scientists and technicians, who wish to participate in structural studies. The NCI R50 award will allow me a firm financial basis on which to extend current studies and initiate new collaborations to include investigators with newly discovered targets or probes but who currently lack the funding to support my collaborative work. Understanding structural and mechanistic aspects of new drug candidates is required to move hit compounds toward leads and a clinical testing funnel, and aid Principal Investigators in obtaining funding for their research, thereby expediting the movement of basic science discoveries from "bench to bedside".