DEVELOPMENT OF NOVEL SMALL MOLECULE INHIBITORS OF AURORA B KINASE SIGNALING

该子项目是利用 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得主要资金， 因此可以在其他CRISP条目中表示。列出的机构是 中心，不一定是研究者的机构。 在细胞分裂期间维持染色体倍性需要染色体分离和胞质分裂的精确协调，使得在所有复制的姐妹染色单体正确地附着于有丝分裂纺锤体并且后期已经开始之前不发生分裂。一个保守的多亚基因子称为染色体乘客复合体（CPC）通过其在有丝分裂早期的动粒和胞质分裂期间的纺锤体中间区的功能在协调有丝分裂和胞质分裂中起着核心作用。它在细胞分裂中的重要作用使得CPC的催化亚基Aurora B激酶成为一个有吸引力的化疗靶点，目前有几种Aurora抑制剂正在临床试验中。这些抑制剂靶向高度保守的ATP结合位点，因此仅显示有限的特异性，从而限制了它们在研究和临床应用中的适用性。与高度保守的催化位点相反，Aurora B与其活化支架INCENP之间的相互作用是这两种蛋白质所特有的，并且代表了潜在的但未开发的干预靶点。在本申请中，我们将利用实验室开发的试剂和测定来筛选Aurora B活性的新型变构干扰物，以用作研究和临床应用的探针。形成本提案特定目标的实验路线将：1）开发高通量、基于流式细胞术的筛选，以鉴定破坏Aurora B/INCENP相互作用的候选小分子; 2）应用化学生物学方法优化这些分子的体外和体内功效。这些实验将与NM-INBre化学生物学和筛选合作核心（CBSC）和UNM分子库筛选中心合作进行，该中心已开发出高通量流式细胞术用于化学库的生化和细胞筛选。从这些研究中，预期不仅将开发用于研究细胞分裂的新型试剂，而且这些试剂将潜在地转化到临床竞技场中，其中靶向该关键结构代表了用于化学治疗干预的极好靶标。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Maintenance of chromosomal ploidy during cell division requires a precise coordination of chromosome segregation and cytokinesis such that the cleavage does not occur before all replicated sister chromatids are correctly attached to the mitotic spindle and anaphase has been initiated. A conserved multi-subunit factor termed the Chromosomal Passenger Complex (CPC) plays a central role in coordinating mitosis and cytokinesis through its functions at the kinetechore early in mitosis and at the spindle midzone during cytokinesis. Its essential role in cell division have made the catalytic subunit of the CPC, Aurora B kinase, an attractive chemotherapeutic target, and there are currently several Aurora inhibitors in clinical trials. These inhibitors target the highly conserved ATP-binding site and consequentially display only limited specificity, thus restricting their applicability for both research- and clinical applications. In contrast to the highly conserved catalytic site, the interaction between Aurora B and its activating scaffold, INCENP, is unique to these two proteins and represents a potential- but unexploited target for intervention. In this application we will take advantage of reagents and assays developed in the laboratory to screen for novel, allosteric disrupters of Aurora B activity for use as probes for research and clinical applications. The lines of experimentation that form the Specific Aims of this proposal will: 1) develop a high-=throughput, flow-cytometry-based screen to identify candidate small molecules that disrupt Aurora B/INCENP interations; and 2) apply a chemical biological approach to optimize the efficacy of these molecules both in vitro and in vivo. These experiments will be performed in collaboration wit the NM-INBre Chemical Biology & Screen Collaborative Core (CBSC), and the UNM Molecular Libraries Screening Center, where high-throughput flow cytometry has been developed for biochemical- and cell-based screening of chemical libraries. From these studies, it is anticipated that not only will novel reagents be developed for the study of cell division, but that these reagents will potentially translate into the clinical arena, where targeting of this critical structure represents an excellent target for chemotherapeutic intervention.