Bivalent Ligands as Chemical Probes to Study Opioid Abuse-enhanced HIV Infection

项目摘要 药物滥用直接导致美国三分之一的艾滋病毒感染。流行病学数据 已经证明阿片类药物滥用是艾滋病毒感染和进展为艾滋病的一个重要风险因素， 越来越多的证据显示μ阿片样物质（莫尔）和CCR 5之间可能存在协同作用 趋化因子受体在这个病理过程中。因此，彻底了解神经通路 可能参与阿片类药物增强艾滋病毒感染是必不可少的。μ阿片样物质的假定二聚化 和CCR 5受体独特地影响免疫细胞功能，并且它们的分子相互作用可能是它们的 在中枢神经系统中有明显的协同作用。二价配体已被证明是用于以下的有力分子工具： G蛋白偶联受体（GPCR）蛋白-蛋白相互作用的表征，以干扰正常的 与这些相互作用相关的功能，甚至通过靶向这种相互作用来治疗疾病。作为证据 概念，我们最近开发的携带MOR-CCR 5双重拮抗剂药效团的二价配体， 在人类巨噬细胞和星形胶质细胞中， 两种拮抗剂的简单混合物。我们的假设是含有两个莫尔的二价配体 拮抗剂和CCR 5拮抗剂可作为化学探针来研究这些受体的相互作用 阿片类药物滥用导致的艾滋病毒感染。我们相信这种配体可以作为 药理学探针，以帮助澄清阿片类药物滥用增强艾滋病毒感染的分子机制， 有助于建立这种不断发展的蛋白质-蛋白质相互作用模型，作为治疗干预的潜在靶点， 阿片类药物滥用增强了神经艾滋病。本课题的具体目标是：1）设计合成新型的 含有莫尔拮抗剂和CCR 5拮抗剂作为双药效团的二价配体， 应用配体结合的莫尔和CCR 5蛋白的晶体结构、分子建模和化学分析， 合成; 2）用受体结合和功能测定表征这些二价配体;和3）检查 二价配体阻断HIV通过CCR 5和CCR 5-莫尔进入和感染的相对功效 交互.我们相信这样的努力将为理解如此复杂的生物学过程奠定基础。 过程，定义一个新的治疗靶点来治疗神经艾滋病，并促进未来的治疗发展， 这种疾病

Project Summary Drug abuse directly contributes to one-third of all HIV infections in the United States. Epidemiological data have demonstrated that opioid abuse is a significant risk factor for HIV infection and progression to AIDS while accumulating evidence reveals possible synergistic interactions between the mu opioid (MOR) and CCR5 chemokine receptors in this pathologic process. Therefore, a thorough understanding of the neural pathways likely involved in opioid enhancement of HIV infection is essential. The putative dimerization of the mu opioid and CCR5 receptors uniquely affects immune cell function and their molecular interactions may underlie their apparently synergistic effects in the CNS. Bivalent ligands have been shown to be powerful molecular tools for characterization of G-protein coupled receptor (GPCR) protein-protein interactions, to interfere with normal function related to these interactions, or even to treat diseases by targeting such interactions. As a proof-of- concept, our recently developed bivalent ligand carrying MOR-CCR5 dual antagonist pharmacophores has shown significantly higher inhibitory effect on HIV-1 invasion in human macrophages and astrocytes compared to a simple mixture of the two antagonists. Our hypothesis is that bivalent ligands containing both an MOR antagonist and a CCR5 antagonist may serve as chemical probes to study the interaction of these receptors with respect to HIV infection enhanced by opioid abuse. We believe a ligand of this kind may serve as a pharmacological probe to help clarify the molecular mechanism of opioid abuse enhanced HIV infection, and help establish this evolving protein-protein interaction model as a potential target for therapeutic intervention in opioid abuse enhanced neuroAIDS. The specific aims of this proposal are to: 1) design and synthesize novel bivalent ligands containing both an MOR antagonist and a CCR5 antagonist as dual pharmacophores by applying crystal structures of ligand bound MOR and CCR5 proteins, molecular modeling, and chemical synthesis; 2) characterize these bivalent ligands with receptor binding and functional assays; and 3) examine the relative efficacy of bivalent ligands in blocking HIV entry and infectivity via CCR5, and CCR5-MOR interactions. We believe such an effort will build the foundation to understand such a complicated biological process, define a novel therapeutic target to treat neuroAIDS, and facilitate future treatment development for the disease.