SYNTHESIS OF ISOTOPE-LABELED CB LIGANDS FOR NMR STUDIES

描述（申请人摘要）： 由于两种大麻素受体亚型（CB 1，CB 2）已被确定， 作为Gi蛋白偶联的七跨膜蛋白的亚组， 受体超家族，一个巨大的努力已被导向了解 CB受体及其配体之间的分子水平相互作用，如 以及设计具有治疗作用的新型大麻模拟物配体， 有用的生物学特性，但没有精神作用， 大麻 然而，许多关于分子水平的未解之谜 受体和大麻模拟配体之间的相互作用， 受体活性位点的结构及其三维结构 仍有待解决。 本研究计划的目的是 获得大麻模拟剂的分子特性的信息， 膜（为了模拟受体环境）通过组合的 使用核磁共振（NMR）和计算机建模方法。 等 信息对于设计新的类似物是至关重要的， 潜在的治疗价值 一个明智的构象性质 所选的一组类似物与三种已知的大麻模拟物基团相关，即 花生四烯基乙醇酰胺（AEA）、氨基烷基吲哚（AAI）和吡唑 （PRZ）将使用NMR方法进行研究，然后使用计算机进行改进 建模方法。 1)将作出特别努力，研究 这些分子在膜环境中的构象。 的3D 膜中配体的结构将使用多种 核磁共振技术，包括脉冲多维核磁共振实验 梯度、转移NOE和旋转回波双共振实验。 2)计算机建模将用于改进NMR确定的 构象 比较分子场分析（CoMFA）和活性 类比法将被应用于检查药效团的定义 并确定活性位点以及绘制受体体积。 的 通过NMR实验和理论计算确定的药效模型 可作为设计新型CB配体和预测 其他未经测试的化合物的生物学行为。 已制定的程序 最终将用于直接研究受体结合的构象 配体在未来。 总的来说，这些研究将提供对 新一代具有增强生物活性的配体的设计 活动

DESCRIPTION (Applicant's Abstract): Since the two cannabinoid receptor subtypes (CB1, CB2) have been identified as a subgroup of the Gi protein-coupled seven-transmembrane-spanning receptor superfamily, a great effort has been directed toward understanding the molecular-level interactions between the CB receptor and its ligand, as well as designing novel cannabimimetic ligands possessing therapeutically useful biological properties but devoid of the psychotropic effects of marijuana. However, many unanswered questions about the molecular-level interactions between the receptor and the cannabimimetic ligands, the nature of the receptor active site(s), and their three-dimensional structures remain to be addressed. The objective of this research proposal is to obtain information on the molecular properties of cannabimimetic agents in membranes (in order to mimic the receptor environment) through the combined use of nuclear magnetic resonance (NMR) and computer modeling methods. Such information is of critical importance for the design of novel analogs of potential therapeutic value. The conformational properties of a judiciously chosen group of analogs related to three known cannabimimetic groups, namely arachidonylethanolamides (AEAs), aminoalkylindoles (AAIs), and pyrazoles (PRZs) will be studied using NMR methods and then refined using computer modeling approaches. 1) A special effort will be made to study the conformations of these molecules in a membrane environment. The 3D structure of the ligand in the membrane will be obtained using a variety of NMR techniques including multidimensional NMR experiments with pulse gradients, Transfer NOE and Rotational Echo Double Resonance experiments. 2) Computer modeling will be used to refine the NMR-determined conformations. Comparative Molecular Field Analysis (CoMFA) and the Active Analog Approach will be applied to examine the definition of pharmacophores and to define the active site as well as to map receptor volume. The pharmacophoric model defined by NMR experiments and theoretical calculations can be used as a guide for designing novel CB ligands and predicting the biological behavior of other untested compounds. The procedures developed will eventually be used to directly study the conformation of receptor-bound ligands in the future. Overall, the studies will provide insights into the design of a new generation of ligands possessing enhanced biological activity.