Heat Shock Protein Function for Drug Discovery

描述(由申请人提供)： 控制肿瘤细胞生长和存活的信号转导途径 为新型抗癌药物的开发提供一个有吸引力的、合理的目标 探员们。调节这些通路的一种特别有效的方法可能是 由改变热休克蛋白(HSP)功能的药物提供 例如Hsp90，而不是任何特定的信号分子。这些 在癌症中，普遍表达的分子伴侣不会发生突变。 相反，它们似乎在促进或促成方面发挥着至关重要的作用 通过增强多重链的稳定性和活性实现肿瘤转化 致癌生长因子受体、激酶和转录因子。完毕 在过去的5年里，已经鉴定了几种小分子天然产物。 作为潜在的抗癌剂，与热休克蛋白或其辅助伴侣相互作用。 其中一种药物，17-N-烯丙氨基-17-去甲氧基格尔达那霉素(17 MG) 进入I期临床试验，但17 mg和其他类别的 到目前为止发现的热休克蛋白结合药物受到严重的限制 临床用途，包括溶解性、新陈代谢、效力差等问题 和/或有限的特异性。实现热休克蛋白新颖化开发的目标 癌症化疗靶点，需要寻找更多具有 在体内调节它们复杂的细胞功能。 最近，一项鉴定和筛选生物活性的专利技术 化合物是由Telik，Inc.开发的，它依赖于分类 根据它们的结合特性，建立一个庞大的化合物文库 (“指纹”)来对抗一组蛋白质。与传统高中形成对比 吞吐量方法，这种与目标相关的亲和性分析(TRAP)方法 允许筛选相对较少数量的纯化合物， 大约200-400，以识别针对特定分子的活性分子 分子靶标或细胞途径。我们假设陷阱的应用 技术对基于记者构造的屏幕生成的数据将允许我们 鉴定新的具有临床应用潜力的热休克蛋白结合化合物 发展。为了验证这一假设，我们汇集了一个多学科的 拥有完成以下任务所需的专业知识和基础设施的团队 以下是具体目标： 1)使用两步法鉴定改变热休克蛋白功能的有效化合物 一系列中等吞吐量、基于细胞的报告分析和陷阱技术。 2)确定最有希望进一步开发的化合物，方法是 量化对细胞增殖、细胞周期进程、细胞凋亡和 特定分子靶点的变化，如erbB2、Akt、突变型P53和 类固醇激素受体。 3)评价新型热激活性化合物的潜在相互作用 用固相药物固定化技术固定特定的热休克蛋白和辅助伴侣 技巧。 在实践层面上，这些探索性研究将提供新的积极的热休克蛋白 可用于进一步验证HSP作为治疗靶点的药物 干预。我们确定的代理也可以作为以下目标的起点 在开发有用的抗癌药物方面进行领先的优化，具有独特的 行动机制。在基础科学层面上，我们的应用程序构成 一种创新的药理学方法来定义 像Hsp90这样的伴侣促进肿瘤的发生。因此，这些见解 在这个项目中取得的成果可能会对 更有效地防治癌症。

DESCRIPTION (provided by applicant): The signal transduction pathways that control tumor cell growth and survival provide an attractive, rational target for the development of new anticancer agents. A particularly effective approach to modulating these pathways could be provided by drugs which alter the function of heat shock proteins (Hsp) such as Hsp90 instead of anyone specific signaling molecule. These ubiquitously expressed molecular chaperones are not mutated in cancer. Instead, they appear to play an essential role in facilitating or enabling neoplastic transformation by enhancing the stability and activity of multiple oncogenic growth factor receptors, kinases and transcription factors. Over the past 5 years, several small molecule natural products have been identified as potential anticancer agents that interact with Hsp or their co-chaperones. One of these agents, 17 -N-allylamino-17 -demethoxygeldanamycin ( 17 MG ) has entered phase I clinical trials, but both 17 MG, and the other classes of Hsp-binding drugs identified so far suffer from serious limitations to their clinical utility including problems with solubility, metabolism, poor potency and/or limited specificity. To achieve the goal of exploiting Hsp as novel cancer chemotherapeutic targets, a need exists to find additional agents with which to modulate their complex cellular functions in vivo. Recently, a proprietary technology for characterizing and screening bioactive compounds has been developed by TELIK, Inc. that relies on the classification of a vast library of compounds according to their binding characteristics ("fingerprints") against a panel of proteins. In contrast to traditional high throughput methods, this Target-Related Affinity Profiling (TRAP) approach allows the screening of a relatively small number of pure compounds, approximately 200-400, to identify active molecules against a specific molecular target or cellular pathway. We hypothesize that application of TRAP technology to data generated by reporter construct-based screens will allow us to identify new Hsp-binding compounds with the potential for clinical development. To test this hypothesis, we have assembled a multidisciplinary team with the expertise and infrastructure required to accomplish the following specific aims: 1) To identify potent compounds that alter Hsp function using a two stage series of moderate throughput, cell-based reporter assays and TRAP technology. 2) To identify the most promising compounds for further development by quantitating effects on proliferation, cell cycle progression, apoptosis and alterations in specific molecular targets such as erbB2, Akt, mutant p53 and steroid hormone receptors. 3) To evaluate the potential interaction of novel heat shock-active compounds with specific Hsp and co-chaperones using solid phase drug immobilization techniques. At a practical level these exploratory studies will provide new Hsp-active agents that can be used to further validate Hsp as targets for therapeutic intervention. The agents we identify can also serve as starting points for lead optimization in the development of useful anticancer agents with unique mechanisms of action. At a basic scientific level our application constitutes an innovative pharmacological approach to defining the mechanisms by which chaperones such as Hsp90 facilitate oncogenesis. Consequently, the insights gained in this project are likely to have significant implications for the more effective prevention and cure of cancer.