ARMET LIGANDS IN ANTI-CANCER DRUG DEVELOPMENT

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 Armet是一种双功能蛋白，存在于所有多细胞动物中。在细胞内，Armet是一个相对未被研究和相对最近的未折叠蛋白质对ER应激的反应的组成部分，我们的工作重点就是Armet的这一方面。 我们正在将人类Armet作为药物开发的靶点，使用非标记筛选来寻找紧密结合的配体，然后对初步筛选的命中进行详细的结合研究，并研究这些选定化合物对细胞的影响。 这项工作的基本原理是，Armet作为处理ER应激的细胞机制的一部分，如果受到紧密结合配体的干扰，将使细胞更容易发生凋亡，这是ER应激的典型结果，但癌细胞通常会避开这种命运。 在KU高通量筛选实验室中，去年春天对10，000种化合物的验证库进行了筛选，其中大部分具有生物活性，这些化合物是在我的实验室中表达和纯化的。我们有一组大约20个来自该屏幕的“命中”，其中几个现在正在我在堪萨斯州立大学的实验室进行积极的研究，在与Armet结合的二级分析中获得了有利的结果。 目前的提案有以下目标：在KU HTS实验室使用即将到达该实验室的ICx技术仪器筛选另外100，000种化合物;通过表面等离子体共振对选定的配体进行详细的结合研究。（KSU的Biacore 3000），（棉酚）结合母体化合物的一组变体;并在HTS实验室和KSU研究最紧密结合的配体对具有广泛恶性特性的人类细胞的影响，特别是寻找使癌细胞易于凋亡的化合物。 反过来，这将导致在未来的工作中，合成一种或多种先导化合物的化学变体，寻求与Armet更紧密的结合，同时保留对癌细胞的预期效果。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Armet is a bifunctional protein apparently present in all multicellular animals. Inside cells, Armet is a relatively unstudied and relatively recently component of the Unfolded Protein Response to ER stress, and it is this aspect of Armet that our work focuses on. We are working on human Armet as a target for drug development, using non-labelled screening to find tight binding ligands, followed by detailed binding studies of hits from the primary screening and studies of the effects of these selected compounds on cells. The rationale underlying this work is that the action of Armet as a part of the cell's machinery for dealing with ER stress, if interfered with by tight-binding ligands, would render cells more susceptible to the apoptosis, a typical result of ER stress, but a fate often circumvented by cancer cells. In the KU High Throughput Screening Laboratory, a validation library of 10,000 compounds, most of which are biologically active, was screened last spring against human Armet, as expressed and purified in my laboratory. We have a group of approximately 20 "hits" from that screen, several of which are under active investigation now in my laboratory at Kansas State University, with favorable results in that secondary analysis of binding to Armet. The current proposal has the following aims: to screen an additional 100,000 compounds at the KU HTS Lab, using an ICx Technology instrument that will soon arrive in that lab; to conduct detailed binding studies of selected ligands by Surface Plasmon Resonance (Biacore 3000 at KSU) and in one case (gossypol) binding of a set of variants of the parent compound; and to examine, both in the HTS Lab and at KSU, the effect of tightest binding ligands on human cells having a wide range of malignant properties, looking specifically for compound that render cancer cells vulnerable to apoptosis. This will lead, in turn, in future work, to synthesis of chemical variants of a lead compound or compounds, seeking tighter binding to Armet while retaining desired effects on cancer cells.