Endocannabinoid Active Sites as Therapeutic Targets

研究及相关-其他项目信息-项目摘要/摘要 该计划项目的目标是探索CB1和CB2大麻素的功能特性 通过开发合适的新型配体并通过获得结构和功能 关于这些受体的信息。在当前周期中，我们已经开发出高亲和力的共价和非亲和力 利用配基辅助蛋白研究共价CB1/CB2配体在受体上的“足迹” 结构(LAPS)方法，包括使用共价配体、受体突变体和LC/MS/MS 方法：研究方法。使用计算得出的受体模型进一步阐述了结果。到目前为止，我们 结果提供了不同类别的大麻能配体相互作用和 通过不同的结合基序激活/去激活CB1和CB2受体 配体-受体构象。在我们的配体和CB1/CB2结构工作的同时，我们的合作者 已经提供了证据表明CB1(Bohn)和CB2(Mackie)不同的激动剂通过 不同的信号通路(功能选择性)。我们假设个别配基- 我们已经确定的受体结合基序反过来可能与不同的可识别信号相关联 导致不同配体相关药理学特征的途径。本节目的一个中心主题 项目续签申请是CB1和CB2中确定的不同信令配置文件将允许我们 设计“功能选择性”配体。这种基于功能药效团的新药物设计应该 导致新的配体具有更好的药理特性。 本计划项目更新建议：1)设计和合成第一代和后代共价和 非共价配体用于探索关键类别的功能性质和结合基序 大麻能配体；2)利用受体突变体和 LC/MS/MS方法；3)确定最成功的配体的CB1和CB2信号谱；以及4)评估 在活体中，CB1和CB2配体使用小鼠四分体测试以及在神经病理性疼痛的动物模型中。 将获得的结果将作为开发成瘾和疼痛治疗方法的基础。

RESEARCH & RELATED - OTHER PROJECT INFORMATION - PROJECT SUMMARY/ABSTRACT The goal of the Program Project is to explore the functional properties of the CB1 and CB2 cannabinoid receptors through the development of suitable novel ligands and by obtaining structural and functional information on these receptors. During the current cycle, we have developed high-affinity covalent and non- covalent CB1/CB2 ligands to study their "foot-printing" at the receptor using a Ligand Assisted Protein Structure (LAPS) approach which includes the use of covalent ligands, receptor mutants and LC/MS/MS methods. The results are further elaborated using computationally-derived receptor models. To date, our results provide initial evidence that different classes of cannabinergic ligands interact and activate/deactivate the CB1 and CB2 receptors through distinct binding motifs associated with distinct ligand-receptor conformations. In parallel with our ligand and CB1/CB2 structural work, our collaborators have provided evidence that CB1 (Bohn) and CB2 (Mackie) different agonists activate these receptors through distinct signaling pathways (functional selectivity). We have hypothesized that the individual ligand- receptor binding motifs we have identified, in turn, may be associated with distinct identifiable signaling pathways leading to different ligand-associated pharmacological profiles. A central theme for this Program Project renewal application is that distinct signaling profiles identified within CB1 and CB2 will allow us to design "functionally selective" ligands. This novel functional pharmacophore-based drug design should lead to new ligands with improved pharmacological profiles. This Program Project renewal proposes to: 1) design and synthesize first and later generation covalent and non-covalent ligands to be used to probe the functional properties and binding motifs of the key classes of cannabinergic ligands; 2) identify the binding motifs of the covalent ligands by using receptor mutants and LC/MS/MS methods; 3) Identify CB1 and CB2 signaling profiles of the most successful ligands; and 4) evaluate in vivo the CB1 and CB2 ligands using the mouse tetrad test as well as in animal models for neuropathic pain. The results to be obtained will serve as a basis for the development for therapies for addiction and pain.