Novel Chemical Probes for Study of 5-HT2R Balance and Function

转化成瘾科学中心（TASC）专注于了解被破坏的 5-羟色胺（5-HT）信号通过5-HT 2A受体（S-HTZAR）和5-HT 2cR在复发中的作用 面对可卡因相关线索时的冲动行为和渴望（线索反应）。的机制 以及5-HT 2AR：5-HT 2CR平衡控制这些行为的神经回路 在项目1和2中进行了探索，积累的数据表明，将5- HT 2AR拮抗剂/5-HT 2cR激动剂特性将被证明特别有效地抑制冲动， 提示反应性并减少复发。我们最近首次证明了5-HT 2A +2CR 在体外和离体中发现了异聚体。这些有趣的数据提出了完全原创的可能性， 两种受体可以直接二聚化并显示出明显不同于 它的各个组成部分。为了探索这些假设，需要新的药理学工具 研究5-HT 2AR：5-HT 2CR信号传导的作用，并阐明5-HT 2 R的潜在功能影响。 二聚化。我们的主要目标是开发新的分子，（1）具有双重活性， 5-HT 2AR拮抗剂和S-HTacR激动剂，和（2）能够进行5-HT 2 R的结构和功能分析 二聚化。项目3采用新的方法设计和合成5-HT 2AR拮抗剂 系于5-HT 2cR激动剂;很少有不同受体配体 被拴在一起，试图刺激一个受体，同时阻断另一个受体。的分子 在本项目中合成的蛋白质将用于研究这些蛋白质的生物学， 与特异性二聚体受体结合并选择性改变其效应途径（项目2，核心B）。 还将合成生物素化和荧光标记的分子，以使位置可视化， 在体外和离体形成受体二聚体。除了合成选择性的二价配体外， Bench是一种体内合成方法，使用受体二聚体作为模板的反应， 探针受体二聚化并提供新的、新的二价配体。实现这些目标将 为我们提供了必要的新颖和创新的药理学工具，以进行详细的分析， 5-HT 2A +2CR异二聚化对5-HT 2 R功能平衡的影响，并测试总体的 假设5-HT 2AR：5-HT 2CR平衡药理学恢复将使有害的 促进可卡因依赖复发的行为。 R E L E V A N C E（参见说明）： 对新工具和治疗开发原理验证的需求至关重要，我们将 将这些努力导向产生具有全新的靶向肾上腺素能分子， 治疗可卡因成瘾的作用机制。

The Translational Addiction Sciences Center (TASC) is focused on understanding the role of disrupted serotonin (5-HT) signaling through the 5-HT2A receptor (S-HTZAR) and 5-HT2cR in relapse precipitated by impulsive behavior and craving in the face of cocaine-associated cues (cue reactivity). The mechanisms and neural circuitry through which the 5-HT2AR:5-HT2CR balance controls these behaviors are being explored in Projects 1 and 2, and accumulating data indicate that strategies incorporating combined 5- HT2AR antagonist/5-HT2cR agonist properties will prove particularly effective to suppress impulsivity and cue reactivity and reduce relapse. We have recently demonstrated for the first time that a 5-HT2A+2CR heteromer is found in vitro and ex vivo. These intriguing data raise the wholly-original possibility that these two receptors may directly dimerize and exhibit biochemical properties that are demonstrably distinct from those of its individual components. To explore these hypotheses, new pharmacological tools are required to study the role of 5-HT2AR:5-HT2CR signaling and to elucidate the potential functional impact of 5-HT2R dimerization. Our primary objectives are to develop novel molecules which (1) possess dual activity as a 5-HT2AR antagonist and S-HTacR agonist, and (2) enable structural and functional analyses of 5-HT2R dimerization. Project 3 is taking the novel approach of designing and synthesizing a 5-HT2AR antagonist tethered to a 5-HT2cR agonist; there have been very few cases in which different receptor ligands have been tethered together in an attempt to stimulate one receptor while blocking the other. The molecules synthesized in this Project will be used to study the biology of these proteins and will have the ability to bind to specific dimeric receptors and alter their effector pathways selectively (Project 2, Core B). Biotinylated and fluorescently-labeled molecules will also be synthesized to visualize the location and formation of receptor dimers in vitro and ex vivo. In addition to synthesizing selective bivalent ligands at the bench, an in vivo synthetic approach, using reactions templated by receptor dimers, will be developed to probe receptor dimerization and provide novel, new bivalent ligands. Achievement of these Aims will provide us with novel and innovative pharmacological tools necessary to conduct detailed analyses of the impact of 5-HT2A+2CR heterodimerization on the balance of 5-HT2R function, and to test the overarching hypothesis that pharmacological restoration of 5-HT2AR:5-HT2CR balance will minimize deleterious behaviors that promote relapse in cocaine dependence. R E L E V A N C E (See instructions): The need for new tools and proof-of-principle for therapeutics development is vitally translational and we will direct these efforts toward the creation of targeted serotonergic molecules with fundamentally new mechanisms of action for treatment of cocaine addiction.