Endocannabinoid Active Sites as Therapeutic Targets

研究及相关-其他项目信息-项目概要/摘要 该计划项目的目标是探索CB 1和CB 2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上的 这些受体的信息。在目前的周期中，我们已经开发了高亲和力的共价和非共价的， 共价CB 1/CB 2配体，使用配体辅助蛋白研究它们在受体上的“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.使用计算衍生的受体模型的结果进一步阐述。迄今为止，我们的 结果提供了初步证据，表明不同类型的大麻能配体相互作用， 通过不同的结合基序激活/失活CB 1和CB 2受体， 配体-受体构象。与我们的配体和CB 1/CB 2结构工作平行，我们的合作者 已经提供了证据表明，CB 1（Bohn）和CB 2（Mackie）不同的激动剂通过以下途径激活这些受体： 不同的信号通路（功能选择性）。我们假设单个配体- 我们已经鉴定的受体结合基序，反过来，可能与不同的可识别的信号传导有关。 导致不同配体相关药理学特征的途径。本方案的一个中心主题是 项目更新应用的一个重要方面是，在CB 1和CB 2中识别出的不同信号特征将使我们能够 设计“功能选择性”配体。这种新型的基于功能性药效团的药物设计应该 从而产生具有改进的药理学特性的新配体。 本项目更新建议：1）设计和合成第一代和第二代共价键， 非共价配体用于探测关键类别的功能特性和结合基序， 大麻能配体; 2）通过使用受体突变体鉴定共价配体的结合基序， LC/MS/MS方法; 3）鉴定最成功的配体的CB 1和CB 2信号传导谱;和4）评估 使用小鼠四分体试验以及在神经性疼痛的动物模型中体内测定CB 1和CB 2配体。 获得的结果将作为开发成瘾和疼痛疗法的基础。

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.