Discovery of Death Ligands Against Cancers

描述(申请人提供)：最近，我们修改了一珠一化合物(OBOC)组合文库方法，在OBOC文库中的每个珠子的表面添加了已知的细胞黏附(或细胞捕获)配体。当活细胞与这种新型的一珠两化合物(OB2C)文库孵育时，面向珠面的捕获细胞的细胞膜暴露在每个珠上显示的库化合物中。有了合适的报告系统，人们应该能够快速检测到能够引起特定生化或细胞反应的珠子(激动剂)。同样，如果细胞受到外源激动剂的刺激，也可以用这种方法发现抑制特定生化或细胞反应的分子(拮抗剂)。在这份R33提案中，我们计划将我们的努力集中在使用新的OB2C技术来发现促凋亡细胞表面作用分子，以对抗血液系统和实体恶性肿瘤。该项目的具体目标如下：目标1：设计和合成OB2C组合文库，用于发现合成的和细胞表面作用的促凋亡分子或死亡配体。目的：筛选OB2C组合文库，寻找抗淋巴癌、急性髓系白血病和实体瘤细胞表面作用的死亡配体。先导化合物将通过聚焦的OB2C组合文库进行进一步优化。目的3：重新合成先导化合物，评价其自身或与肿瘤细胞表面靶向配体偶联后的促凋亡功能。这些促凋亡剂的作用机制将被确定。影响：超高吞吐量OB2C文库方法高效经济。一旦优化，许多学术研究人员就可以很容易地将其应用于他们的研究。在这项拟议的研究中将开发的死亡配体可以开发成新的有效但毒性较低的癌症治疗药物。这些配体还可以用作基础癌症生物学和系统生物学研究的生物活性探针，特别是在它们的作用机制被阐明之后。

DESCRIPTION (provided by applicant): Recently, we have modified the one-bead-one-compound (OBOC) combinatorial library method by adding a known cell adhesion (or cell capturing) ligand to the surface of every bead in the OBOC library. When live cells are incubated with such novel one-bead-two-compound (OB2C) libraries, the cell membranes of the captured cells facing the bead surface are exposed to the library compounds displayed on each bead. With an appropriate reporter system, one should be able to rapidly detect beads that can elicit a specific biochemical or cellular response (agonists). Similarly, if the cells are stimulatd by an exogenous agonist, molecules that suppress specific biochemical or cellular response (antagonists) can also be discovered with this approach. In this R33 proposal, we plan to focus our effort on using the novel OB2C technology to discover pro- apoptotic cell surface acting molecules against both hematologic and solid malignancies. Specific aims of this proposed project are as follows: Aim 1: To design and synthesize OB2C combinatorial libraries for the discovery of synthetic and cell surface acting pro-apoptotic molecules or death ligands. Aim 2: To screen OB2C combinatorial libraries for the discovery of synthetic and cell surface acting death ligands against lymphoid cancer, acute myeloid leukemia and solid tumor cells. The lead compounds will be further optimized with focused OB2C combinatorial libraries. Aim 3: To resynthesize the lead compounds and evaluate their pro-apoptotic functions by themselves or after conjugation to the cancer cell surface targeting ligands. The mechanisms of action of these pro-apoptotic agents will be determined. Impact: The ultra-high throughput OB2C library method is highly efficient and economical. Once optimized, it can be readily applied by many academic investigators to their research. The death ligands to be developed in this proposed research can be developed into novel effective but less toxic cancer therapeutics. These ligands may also be used as biologically active probes for basic cancer biology and systems biology research, particularly after their mechanisms of action have been elucidated.