Glutamate Vesicular Transport:Pharmacophore Development

谷氨酸囊泡运输：药效团开发

• 批准号: 6724779
• 负责人: charles mark thompson
• 金额: $ 33.25万
• 依托单位: UNIVERSITY OF MONTANA
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-09-01 至 2007-03-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6724779
• 关键词: affinity labeling; chemical synthesis; drug design /synthesis /production; drug receptors; excitatory aminoacid; glutamate receptor; glutamate transporter; high performance liquid chromatography; high throughput technology; infrared spectrometry; laboratory rat; mass spectrometry; model design /development; molecular site; neurotransmitter antagonist; nuclear magnetic resonance spectroscopy; physical model; protein structure function; quinoline; receptor binding; synaptic vesicles; thin layer chromatography; ultraviolet spectrometry

DESCRIPTION (provided by applicant): The primary goals of this competing renewal application are to define the structural requirements for selective and potent binding to the glutamate synaptic vesicular transporter (VGLUT) and to advance our understanding of the structural and functional roles of VGLUT. The long-range goals of this study are to: (a) develop pharmacophore and structural models of VGLUT that will aid the development of selective molecules that can regulate vesicular storage, uptake and release of glutamate and, (b) define the role and contribution of VGLUTs and related vesicular transporter proteins in vesicle function. The proposed strategy will optimize VGLUT inhibition and binding selectivity using results from our current project combined with novel chemical, computational, biochemical and pharmacological approaches. A concurrent strategy will seek to obtain a more global understanding of VGLUT structure and function. Using integrative strategies, we will increase our project activity and broaden our efforts to define the structural, functional and pharmacological roles of VGLUT. For the proposed grant period, we will address the following objectives: Specific Aim 1: Design and synthesize inhibitor structures that selectively bind VGLUT. Emphasis will be placed on developing inhibitors that satisfy the elements of a dynamic pharmacophore model, for example, a cyclic platform with regio- and stereochemically defined placement of acid and lipophilic substituents. Specific Aim 2: Test the activity of compounds prepared in SA 1 as inhibitors of VGLUT. The ability of these compounds to bind EAA receptors and cellular transporters also will be determined to assess the specificity at VGLUT. Specific Aim 3: Generate a computationally-derived pharmacophore model of VGLUT utilizing the chemical, biochemical and pharmacological data generated in SA's 1 & 2, The pharmacophore model will be used to refine and iterate inhibitor design (SA1), affinity agents (SA4), protein probe structures (SA's 4 and 5), and to create protein 3D structural hypotheses consistent with the probe and cross-linking results (SA4 & 5). Specific Aim 4: Characterize the transmembrane domain regions of VGLUT and neighboring architectures that contribute to VGLUT structure and function. Using N-term VGLUT antibodies, affinity tags, tethered- and cross-linking probes, VGLUT residues and structures proximal to VGLUT will be isolated and identified by mass spectrometry.

描述（由申请人提供）：这种竞争性更新应用的主要目标是定义与谷氨酸突触囊泡转运蛋白（VGLUT）的选择性和有效结合的结构要求，并促进我们对VGLUT的结构和功能作用的理解。这项研究的远距离目标是：（a）开发VGLUT的药理和结构模型，这些模型将有助于开发选择性分子，这些分子可以调节水泡储存，摄取和谷氨酸的摄取和释放，以及（（b）定义vglut和vglut和相关的Vesicular Transporter蛋白在vesicle函数中的作用和贡献。 提出的策略将使用我们当前项目的结果与新型化学，计算，生化和药理学方法相结合，优化VGLUT抑制和结合选择性。并发策略将寻求对VGLUT结构和功能的更全球化的理解。使用综合策略，我们将增加项目活动，并扩大我们定义VGLUT的结构，功能和药理作用的努力。在拟议的赠款期间，我们将解决以下目标： 特定目标1：设计和合成抑制剂结构，这些抑制剂结构有选择地结合VGLUT。重点将放在开发满足动态药效团模型元素的抑制剂上，例如，具有区域和立体定义的酸和亲脂性取代基的循环平台。 具体目标2：测试Sa 1中作为VGLUT抑制剂制备的化合物的活性。这些化合物结合EAA受体和细胞转运蛋白的能力也将确定以评估VGLUT的特异性。 具体目的3：生成一种使用化学，生化和药理学数据的计算衍生的vglut药效团模型，在SA的1和2中产生的药理数据将使用药效团模型来完善和迭代抑制剂设计（SA1），亲和力（SA4），亲和力（SA4），蛋白质探针结构（SA的4和5）和蛋白质结构（SA的4和5）和蛋白质（SA的4和5），以及蛋白质3D，3D。交联结果（SA4和5）。 特定目标4：表征有助于VGLUT结构和功能的VGLUT和相邻体系结构的跨膜结构域区域。使用N-期限VGLUT抗体，亲和力标签，束缚和交联探针，近端与VGLUT的VGLUT残基和结构通过质谱分离和鉴定。