Target identification of small molecule LDN/OSU-215111

小分子LDN/OSU-215111的靶点鉴定

• 批准号: 9885609
• 负责人: CHIEN-LIANG GLENN LIN
• 金额: $ 45.68万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-03-01 至 2023-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9885609
• 关键词: Activities of Daily Living; Affinity; Alzheimer' s Disease; Alzheimer' s disease model; Alzheimer' s disease patient; Astrocytes; Binding; Cognitive; Disease; Event; Excitatory Amino Acid Transporter 2; Glutamate Transporter; Glutamates; Goals; In Vitro; Lead; Maintenance; Mediating; Memory; Messenger RNA; Nerve Degeneration; Neurofibrillary Tangles; Pathogenesis; Pathology; Phenotype; Play; Process; Protein Microchips; Proteins; Regulation; Role; Senile Plaques; Series; Structure; Synapses; System; Translational Activation; Translations; clinical candidate; cognitive function; drug discovery; extracellular; glial activation; improved; knock-down; mouse model; novel; novel therapeutics; pre-clinical; preclinical development; protein function; small molecule; tau phosphorylation

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.

谷氨酸转运蛋白EAAT 2在细胞外环境的稳态调节中起着关键作用。 谷氨酸水平EAAT 2在认知记忆功能中也起着重要作用。然而，损失 EAAT 2蛋白和功能在阿尔茨海默病（AD）患者中常见。我们有 发现并开发了一系列新型小分子，可以通过以下方式增加EAAT 2表达 一种新的翻译激活机制。这些分子已经被证明能够 正常化谷氨酸稳态失调并在两种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。但是我们最初的铅化合物 是用表型方法鉴定的。目前我们还不知道复合目标的 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，我们将利用串联的亲和探针下拉方法和蛋白质 微阵列方法来鉴定推定的化合物靶标。在目标2中，我们将调查已确定的 候选人，并确定负责LDN/OSU-215111介导的靶蛋白 EAAT 2诱导。我们将首先验证LDN/OSU-215111与鉴定的蛋白质的相互作用 体外一旦相互作用得到证实，我们将研究是否敲低候选蛋白质 导致LDN/OSU-215111介导的EAAT 2诱导丧失。目标是确定 复合目标