COMBINATORIAL CHEMISTRY APPROACHES TO TARGETING CELL CYCLE REGULATORS

项目1的主要目标将是开发产生生物活性分子结构多样性的新方法，并将这些化合物文库定向到细胞周期控制途径中的关键靶点。我们的首要主题将是从生物学靶标的知识或从噬菌体展示文库的筛选中鉴定出的多肽序列或蛋白质结构域，并通过系统地应用新的组合方法，将它们还原为高效、生物可用和体内活性的抗肿瘤药物。为此，我们将开发几种方法来方便地建立潜在活性化合物的大型组合文库，然后将每一种方法应用于鉴定用于干扰细胞周期调控中的关键步骤的前导结构；受体酪氨酸激酶(CDK)功能、信号转导和转录激活因子(STAT)信号和细胞周期蛋白依赖激酶(CDK)活性。我们对RTK抑制剂的方法将采用两种策略，涉及蛋白质表面结合剂的设计和2-苯基亚硫基吲哚小杂环衍生物文库的构建。在STAT领域，我们将使用固相平行合成来制备阻断STAT3的SH-2结构域的多肽模拟物文库。通过CDK，我们将开发天然抑制剂p16和p21表面关键螺旋结构域的模拟库，以及与ATP竞争结合到酶活性部位的小杂环衍生物。我们将与项目2、3和4合作，针对细胞周期信号通路中的这些关键步骤筛选文库。我们的主要重点将是鉴定和优化RTKs(包括血小板衍生生长因子受体、表皮生长因子受体和ErbB2)、STAT蛋白家族(特别是STAT3)和三种主要的细胞周期蛋白依赖激酶(CDK2、CDK4和CDK6)的新抑制剂。

The primary aims of project 1 will be to develop new approaches for generating structural diversity in bioactive molecules and to target these compound libraries to critical targets in cell cycle control pathways. Our overriding theme will be to take peptide sequences or protein domains that have been identified either from knowledge of the biological target or from screening of phage display libraries and, by systematic application of novel combinatorial methods, to reduce them to highly potent, bioavailable and in vivo active anti-tumor agents. To do this we will develop several approaches to the facile generation of large combinatorial libraries of potentially active compounds and then apply each of these to the identification of lead structures for disruption of thee key steps in cell cycle control; receptor tyrosine kinase (CDK) function, signal transducers, and activators of transcription (STAT) signaling and cyclin dependent kinase (CDK) activity. Our approach to RTK inhibitors will exploit two strategies involving the design of protein surface binding agents and the construction of libraries of small heterocyclic derivatives of 2-phenylsulfenylindole. In the STAT area we will use solid phase parallel synthesis to prepare libraries of peptidomimetics that block the SH-2 domain of STAT3. With the CDKs we will develop libraries of mimics of key helical domains on the surface of the natural inhibitors, p16 and p21, as well as small heterocyclic derivatives that compete with ATP for binding to the enzyme active site. Screening of the libraries against these key steps in the cell cycle signaling pathways will be carried out in collaboration with Projects 2, 3, and 4. Our primary focus will be the identification and optimization of novel inhibitors for RTKs (including platelet derived growth factor receptor, epidermal growth factor receptor and ErbB2), the STAT family of proteins (with particular focus on STAT3) and the three principal cyclin dependent kinases (CDK2, CDK4 and CDK6).