DESIGN OF LIGANDS SELECTIVE FOR THE DAG SUPERFAMILY

DAG超家族选择性调节剂的发现 细胞中的佛波酯受体，目前包括PKC， 嵌合体和UNC-13对于确定 生理过程中的个别蛋白质。这样的发现 DAG超家族的小分子调节剂在很大程度上是一种未被满足的 药理上的需要。这项提案的主要目标是创造 针对特定DAG家族成员的新型非肿瘤促进配体。 目前，我们知道配体如何与DAG位点结合，我们知道如何 可以对配体进行修饰以实现DAG之间的选择性 超级家庭成员。改良型DAG位点调节子的发现 选择性不仅有助于阐明 生物过程中的单个蛋白质，但最终应该 促进发现治疗疾病的新疗法 癌症和阿尔茨海默氏症等疾病。Bryostatin提供了一种 DAG超家族调制器的一个引人注目的例子已经找到了用途 临床上用于治疗癌症。在这项研究过程中， 我们将实现以下目标： .1.ILV、ITS相互作用的分子模拟研究 苯唑内坦模拟和选择PKC CRD内的合成靶点， 激活剂结合域，以及 将进行嵌合体和UNC-13。这些研究将指导 可能显示DAG位点的新合成靶点的设计和选择 选择性。将开展结构-活性关系研究 揭示对结合亲和力和 选择性，从而指导新的合成结构的设计。 .2.加成苯并内酰胺类化合物的合成 可以增强同工酶选择性的取代基将是 使用我们已经开发的化学方法进行的。 同时，我们还将探索新型火员环的活性。 ILV的类似物[2-吡咯烷酮]；初步实验显示 化合物与PKCalpha具有良好的亲和力。 .3.测定新合成的DAG家族的选择性 材料，体外结合研究将使用 包括突变体在内的相关重组PKC同工酶以及 嵌合体和UNC-13。以评估改变的影响 配体疏水侧链对其促肿瘤活性的研究 将进行皮肤增生症的治疗。

The discovery of selective modulators of the DAG superfamily, the major phorbel ester receptors in cells, which presently includes PKC, the chimaerins, and unc-13 is crucial to ascertaining the role of the individual proteins in physiological processes. The discovery of such small molecule modulators of the DAG superfamily is largely an unmet pharmacological need. The major objective of this proposal is to create novel, non-tumor promoting ligands for specific DAG family members. Presently, we know how ligands bind to the DAG site, and we know how ligands might be modified to achieve selectivity among the DAG superfamily members. The discovery of DAG-site modulators of improve selectivity will not only aid the elucidation of the roles of the individual proteins in biological processes, but should eventually foster the discovery of novel therapeutics for the treatment of disease states such as cancer and Alzheimer's disease. Bryostatin provides a compelling example of a DAG superfamily modulator that has found use clinically in the treatment of cancer. During the course of this study, we will undertake the following aims: .1. Molecular modeling studies of the interactions of ILV, its benzolactam mimic, and selected synthetic targets within the PKC CRDs, the activator-binding domains, as well as the DAG binding site of the chimaerins and unc-13 will be conducted. These studies will guide the design and selection of new synthetic targets likely to exhibit DAG-site selectivity. Structure-activity relationship studies will be carried out to uncover structural features important to both binding affinity and selectivity, thus guiding the design of novel structures of synthesis. .2. The synthesis of additional benzolactam analogues bearing substituents that may confer enhanced isozyme selectivity will be carried out using chemical methods already developed by us. Simultaneously, we will explore the activity of novel fire-membered ring analogues [2-pyrrolidones] of ILV; preliminary experiments reveal these compounds to bind with good affinity to PKCalpha. .3. To determine the DAG family selectivity of the newly synthesized materials, in vitro binding studies will be carried out using the relevant recombinant PKC isozymes including mutants as well as the chimaerins and unc-13. To assess the effect of alterations in the ligand's hydrophobic side chain on its tumor promoting activity, studies of skin hyperplasia will be conducted.