Discovery of Death Ligands Against Cancers

描述（由申请人提供）：最近，我们通过在OBOC库中的每个头的表面添加已知的细胞粘附（或细胞捕获）配体，改进了一头一化合物（OBOC）组合文库方法。当活细胞与这种新型的单头双化合物（OB2C）文库一起孵育时，捕获的细胞的细胞膜面向珠表面，暴露在每个珠上显示的文库化合物中。有了适当的报告系统，人们应该能够快速检测到可以引起特定生化或细胞反应的珠子（激动剂）。同样，如果细胞受到外源性激动剂的刺激，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）。在这项R33提案中，我们计划将重点放在使用新的OB2C技术来发现促凋亡细胞表面作用于血液和实体恶性肿瘤的分子上。本课题的具体目的如下：目的1：设计和合成OB2C组合文库，用于发现合成和细胞表面作用的促凋亡分子或死亡配体。目的2：筛选OB2C组合文库，以发现针对淋巴细胞癌、急性髓系白血病和实体肿瘤细胞的合成和细胞表面作用的死亡配体。先导化合物将通过重点OB2C组合库进一步优化。目的3：重新合成先导化合物，并评价其本身或结合到癌细胞表面靶向配体后的促凋亡功能。这些促凋亡药物的作用机制将被确定。影响：超高吞吐量OB2C库方法高效、经济。一旦优化，它可以很容易地被许多学术研究者应用到他们的研究中。本研究拟开发的死亡配体可发展为新型有效且毒性较低的癌症治疗药物。这些配体也可以作为生物活性探针用于基础癌症生物学和系统生物学的研究，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.