Opiate Bivalent Ligands:Structure/Function Studies

描述(由申请人提供)：有越来越多的证据表明至少 阿片类药物的某些药理作用是通过阿片类药物介导的 以二聚体或寡聚体形式组织的受体。此外，它现在是 在某些情况下，不同类型的阿片受体可以关联 相互作用形成杂二聚体。异二聚体可能会 调节与介导的信号转导不同的信号转导途径 通过单体或同二聚体的阿片受体是一种有趣的可能性 可能与耐受性和身体依赖有关。出于这些原因， 该项目的广泛、长期目标是调查阿片类药物的作用 通过多学科、协调的方法进行受体二聚化。在……里面 项目1，将设计和合成药理工具，以确定 Mu-Delta阿片受体二聚体的存在。设计方法涉及到 同时占用相邻的MU和Delta识别位置 通过含有MU激动剂和Delta的单一二价配体形成的异二聚体 拮抗剂药效团将对绑定和功能进行优化 通过改变连接药效团的间隔物的长度来实现。 在项目2中，u和增量阿片受体之间的相互作用将是 在培养细胞中进行了研究。Mu受体的活动将是 检测不同水平表达的Delta受体。的能力 MU选择性阿片激动剂抑制细胞内cAMP或 对ERK1/2活性的刺激将在表达时确定 三角洲阿片受体处于激活或失活状态。慕慕大的能力 阿片激动剂诱导细胞适应性反应，如脱敏 或在MU阿片受体激活期间受体内化或 将检查失活情况。项目3涉及分子模拟 不同二聚体界面中的不同二聚体结构。每一座都将被建造 并根据从蛋白质中提取的理论评分函数进行评估 同源和长时间尺度分子动力学计算。二价配体 已被实验发现在阿片类药物识别部位之间架起桥梁 阿片受体二聚体将被用作分子尺子，为 关于结合位点之间距离的信息。长期测试 目标化合物将在小鼠身上进行，以确定是否在体内存在 与项目1和2中获得的体外数据相关联。

DESCRIPTION (provided by applicant): There is burgeoning evidence that at least some of the pharmacological effects of opioids are mediated via opioid receptors that are organized as dimers or oligomers. Moreover, it is now established that in some cases different types opioid receptors can associate with one another to form heterodimers. The possibility that heterodimers may modulate signal transduction pathways that are not identical to those mediated by monomeric or homodimeric opioid receptors is an intriguing possibility that may have a bearing on tolerance and physical dependence. For these reasons, the broad, long-term objective of this project is to investigate the role of opioid receptor dimerization through a multidisciplinary, coordinated approach. In project 1, pharmacologic tools will be designed and synthesized to identify the presence of mu-delta opioid receptor dimers. The design approach involves the simultaneous occupation of neighboring mu and delta recognition sites in a heterodimer by a single bivalent ligand that contains mu agonist and delta antagonist pharmacophores. Optimization of binding and function will be accomplished by varying the length of the spacer that links the pharmacophores. In project 2, the interaction between mu and delta opioid receptors will be investigated in cultured cells. The activities of the mu receptors will be examined at different levels of expressed delta receptors. The ability of mu-selective opioid agonists to inhibit the production of intracellular CAMP or stimulation of the ERK1/2 activities will be determined when the expressed delta opioid receptors are being activated or inactivated. The ability of mu opioid agonists to elicit cellular adaptive responses such as desensitization or receptor internalization during mu opioid receptor activation or inactivation will be examined. Project 3 involves the molecular simulation of different dimer structures in a variety of dimer interfaces. Each will be built and evaluated according to theoretical scoring functions taken from protein homology and long time-scale molecular dynamics calculations. Bivalent ligands that have been found experimentally to bridge the opioid recognition sites in opioid receptor dimers will be employed as "molecular rulers" to provide information on the distance between binding sites. Chronic testing of the target compounds will be carried out in mice to determine if there are in vivo correlates with the in vitro data obtained in projects 1 and 2.