Target identification of small molecule LDN/OSU-215111

谷氨酸转运体EAAT2在胞外动态平衡调节中的重要作用 谷氨酸水平。EAAT2在认知记忆功能中也起着至关重要的作用。然而，损失 EAAT2蛋白及其功能常见于阿尔茨海默病(AD)患者。我们有 发现并开发了一系列新的小分子，可以通过 一种新的翻译激活机制。这些分子已被证明能够 使谷氨酸代谢紊乱正常化，并在两种AD小鼠模型中提供显著益处。 该项目目前处于临床前开发阶段。然而，我们最初的先导化合物 用表型方法进行了鉴定。在这一点上，我们仍然不知道复合目标。这个 本研究的重点是确定LDN/OSU-215111的靶蛋白，这是我们的临床前研究 候选人。在目标1中，我们将同时使用亲和探针下拉方法和一种蛋白质 微阵列技术用于鉴定可能的化合物靶标。在目标2中，我们将调查已确认的 候选并确定负责低密度脂蛋白/OSU-215111介导的靶蛋白 EAAT2诱导。我们将首先验证LDN/OSU-215111与鉴定的蛋白质的相互作用 在试管中。一旦相互作用得到确认，我们将调查是否击倒候选蛋白质 结果失去了LDN/OSU-215111介导的EAAT2诱导。目标是确定 复合目标。

Glutamate transporter EAAT2 plays a critical role in the homeostatic regulation of extracellular glutamate levels. EAAT2 also plays an essential role in cognitive memory functions. However, loss of EAAT2 protein and function is commonly found in Alzheimer’s Disease (AD) patients. We have discovered and developed a novel series of small molecules that can increase EAAT2 expression via a novel translational activation mechanism. These molecules have been proved to be capable of normalizing glutamate dyshomeostasis and providing significant benefits in two mouse models of AD. This project is currently at the pre-clinical development stage. However, our original lead compound was identified with a phenotypic approach. At this point, we still don't know the compound target. The focus of this study is to identify the target protein of LDN/OSU-215111, which is our pre-clinical candidate. In Aim 1, we will utilize, in tandem, an affinity probe pull-down approach and a protein microarray approach to identify putative compound targets. In Aim 2, We will investigate the identified candidates and determine the target protein that is responsible for LDN/OSU-215111-mediated EAAT2 induction. We will first validate the interaction of LDN/OSU-215111 with the identified proteins in vitro. Once the interaction is confirmed, we will investigate if knockdown of the candidate protein results in loss of LDN/OSU-215111-mediated EAAT2 induction. The goal is to determine the compound target.