NOVEL MOLECULAR SITE FOR ANTIDOPAMINERGIC EFFECTS

具有抗多巴胺能作用的新型分子位点

• 批准号: 3378828
• 负责人: Richard B Mailman
• 金额: $ 15.41万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 1985
• 资助国家: 美国
• 起止时间: 1985-04-01 至 1992-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3378828
• 关键词: adenylate cyclase; affinity chromatography; amygdala; antiserum; autoradiography; brain metabolism; chemical binding; chemical structure function; computer graphics /printing; corpus striatum; cyclic AMP; drug design /synthesis /production; high performance liquid chromatography; immunoelectron microscopy; neural transmission; psychopharmacology; radiotracer; tranquilizer; tritium

These neuropharmacological studies application will study the biochemical and molecular mechanisms by which the D1 subclass of dopamine receptors cause their psychopharmacological effects. Several hypotheses will be tested: that there is biochemical heterogeneity of D1 receptors; that only a subset of these receptors are linked biochemically to cAMP synthesis; that a subset of these D1 dopamine receptors interact functionally with D2 receptors as part of the same multimolecular complex; and that it is possible to model this receptor class at the molecular level and to use such models to design drugs that have selectivity for certain subpopulations of D1 receptors. These hypotheses will be tested by studying the basis for apparent multiplicity of D1-like receptors, e.g., by comparing the occurrence of [3H]-SCH23390 binding sites versus dopamine-sensitive adenylate cyclase activity (DA-ACase) or the potency of selected drugs to cause dopaminergically-mediated behaviors. Lesioning and pharmacological studies will be used to compare limbic areas to striatum on the basis of these functional and receptor characteristics. Several series of rigid and semi-rigid dopamine agonists and antagonists will be compared using both in vivo and in vitro pharmacological methods. These data will be used in computer-assisted molecular modeling studies to model the active site of D1 receptors. Although initially these studies will assume a single receptor, it is assumed that the biological studies ultimately will provide data permitting at least two types of sites (e.g., one linked to stimulation of adenylate cyclase, and one not) to be modeled and defined. The molecular modeling studies will be modified continually to incorporate the data from receptor solubilization, purification, and characterization studies which will be a major emphasis of this research. As a consequence of our demonstration that SCH23390 binds tenaciously to its physiologically important receptor(s), the purification experiments will rely heavily on affinity chromatography using a 4-alkylphenyl-substituted analog of SCH23390 that we will synthesize. The availability of purified or partially purified receptor(s) will lead to raising of antibodies against these proteins, and to the initiation of molecular biological study of these proteins. We will perform immunohistochemical localization of these receptors, and also conduct immunoneutralization studies. The molecular biological studies will be aimed at the physiological mechanisms involved in expression and regulation of D1 receptors (e.g., synthesis, posttranslational modifications, sites of expression, etc.).

这些神经药理学研究应用将研究生物化学 以及多巴胺受体D1亚类 导致了它们的精神药理作用。 几个假设将是 测试：D1受体存在生物化学异质性;仅 这些受体的一个子集与cAMP合成生物化学连接; 这些D1多巴胺受体的一个子集在功能上与D2 受体作为同一个多分子复合物的一部分;它是 有可能在分子水平上模拟这种受体， 这些模型设计出的药物对某些 D1受体的亚群。 这些假设将由以下人员进行检验： 研究D1样受体表观多样性的基础，例如，通过 比较[3 H]-SCH 23390结合位点与 多巴胺敏感性腺苷酸环化酶活性（DA-ACase）或 选择药物来引起多巴胺能介导的行为。 损伤和 药理学研究将用于比较边缘区和纹状体， 这些功能和受体特征的基础。 几个系列 将比较刚性和半刚性多巴胺激动剂和拮抗剂 使用体内和体外药理学方法。 这些数据将 用于计算机辅助分子建模研究，以模拟活性 D1受体。 虽然这些研究最初假设 单受体，假设生物学研究最终将 提供允许至少两种类型的站点的数据（例如，一个连接至 刺激腺苷酸环化酶，和一个没有）进行建模和定义。 分子模拟研究将不断修改， 来自受体溶解、纯化和表征的数据 这将是本研究的重点。 因此 我们证明SCH 23390与其生理学上的 重要的受体，纯化实验将严重依赖于 使用4-烷基苯基取代的类似物进行亲和层析 SCH 23390，我们将合成。 纯化或 部分纯化的受体将导致产生抗 这些蛋白质，并启动分子生物学研究， 这些蛋白质。 我们将进行免疫组织化学定位这些 受体，并进行免疫中和研究。 分子 生物学研究的目标将是有关的生理机制 在D1受体的表达和调节中（例如，合成， 翻译后修饰、表达位点等）。