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

项目摘要 药物滥用直接导致了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发展为艾滋病的一个重要风险因素，而 越来越多的证据揭示了Mu阿片类药物(MOR)和CCR5之间可能的协同作用 趋化因子受体参与这一病理过程。因此，对神经通路的彻底了解 可能参与阿片类药物对艾滋病毒感染的增强是必不可少的。Mu阿片类药物的二聚化反应 CCR5受体独一无二地影响免疫细胞功能，它们的分子相互作用可能是其 中枢神经系统有明显的协同效应。二价配体已被证明是一种强大的分子工具 G蛋白偶联受体(GPCR)蛋白-蛋白相互作用的特征，以干扰正常 与这些相互作用相关的功能，甚至通过靶向这些相互作用来治疗疾病。作为证明- 概念，我们最近开发的携带MOR-CCR5双拮抗剂药效团的二价配体 与人巨噬细胞和星形胶质细胞相比，对HIV-1侵袭的抑制作用显著增强 两个对手的简单混合体。我们的假设是，含有两个MOR的二价配体 拮抗剂和CCR5拮抗剂可作为研究这些受体相互作用的化学探针。 关于因滥用阿片类药物而加剧的艾滋病毒感染。我们认为这种配体可以作为一种 有助于阐明阿片类药物滥用加剧艾滋病毒感染的分子机制的药理学探测，以及 帮助建立这种不断进化的蛋白质相互作用模型，作为治疗干预的潜在靶点 阿片类药物滥用加剧了神经艾滋病。这项提议的具体目的是：1)设计和合成小说 同时含有MOR拮抗剂和CCR5拮抗剂作为双重药效团的二价配体 应用配体结合的MOR和CCR5蛋白的晶体结构、分子模拟和化学 合成；2)用受体结合和功能分析表征这些二价配体；以及3)检查 二价配体通过CCR5和CCR5-MOR阻断HIV进入和传染性的相对有效性 互动。我们相信这样的努力将为理解如此复杂的生物学奠定基础。 过程，确定治疗神经艾滋病的新的治疗目标，并促进未来的治疗开发 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.