PHARMACOCHEMICAL STUDIES OF OPIATE NARCOTICS

描述（来自申请人摘要）： 我们继续对阿片类药物进行跨学科研究的总体目标是： 减少呼吸的行为选择性镇痛药的设计 抑郁和身体依赖倾向。我们的工作假设 所有提出的研究都是， 三种克隆的阿片受体（μ、δ和κ），但具有不同的 三种受体的激活和抑制特性的组合 可能是很有前途的行为选择剂为了确定这些候选人， 两大主题将继续：1）继续在体外和体内 活化选择性配体的表征。这些研究的目的是 为了鉴定在所述浓度下激动剂和拮抗剂活性的特定组合， 三个克隆受体，导致镇痛与减少呼吸 抑郁和奖励反应。2)继续描述 在每个克隆的阿片样物质上识别和激活的要求 受体（μ、δ和κ）。这些研究的目的是设计新的 具有所需的激活和抑制组合的治疗剂 三种受体的特性。在这些跨学科的努力中， 将在转染的细胞中进行亲和力和活化的体外评估。 含有克隆的小鼠μ、δ和κ受体的细胞。所有体内 将使用野生型和相应的Mu、Delta和 三种阿片类药物基因都有功能障碍kappa“敲除”小鼠 受体。选择用于体外评估的配体将是具有以下结构的那些： 活化选择性的现有证据。选择用于体内的配体 评估将包括那些在以下条件下已验证活化选择性的物质： 转染细胞研究。识别和识别的所有分子要求 通过计算方法开发的受体的激活将用于 克隆的小鼠μ、δ和κ受体。具体来说，普通分子 识别所有三种受体的决定因素， 将获得它们中的每一个的激活的决定因素。机械论 激活要求的含义将通过模拟配体来探讨 受体与我们构建的阿片受体3D模型的相互作用 实验室从这些研究中确定的重要残基突变 将进行活化并测定其药理作用。 使用3D模型明确表征配体受体复合物 将弥合结构和功能之间的差距， 单独研究配体。以小鼠及其三个克隆为重点 受体将最大限度地提高所获得的结果的一致性， 机械论和治疗论的推论。

DESCRIPTION (from applicant's abstract): The overall goal of our continuing interdisciplinary studies of opioids is the design of behaviorally selective analgesics with diminished respiratory depression and physical dependence liability. The working hypothesis underlying all the proposed studies is that ligands that bind with significant affinity to the three cloned opioid receptors (mu, delta and kappa) but with different combinations of activation and inhibition properties at the three receptors could be promising behaviorally selective agents. To identify such candidates, two major themes will be pursued: 1) continued in vitro and in vivo characterization of activation selective ligands. The goal of these studies is to identify the specific combinations of agonist and antagonist activity at the three cloned receptors that lead to analgesia with diminished respiratory depression and reward response. 2) continued characterization of the requirements of recognition and activation at each of the cloned opioid receptors (mu, delta and kappa). The goal of these studies is to design new therapeutic agents with the desired combinations of activation and inhibition properties at the three receptors. In these interdisciplinary efforts, all in vitro assessment of affinities and activation will be performed in transfected cells containing cloned mouse, mu, delta and kappa receptors. All in vivo studies will be carried out using wild type and the corresponding mu, delta and kappa "knockout" mice with dysfunctinal genes for each of the three opioid receptors. Ligands selected for the in vitro assessment will be those with existing evidence for activation selectivity. Ligands selected for in vivo assessment will those for which activation selectivity has been verified in the transfected cells studies. All molecular requirements for recognition and activation of receptors developed by computational methods will be made for the cloned mouse mu, delta and kappa receptors. Specifically, common molecular determinants for recognition of all three receptors and possibly unique determinants of activation of each of them will be obtained. The mechanistic implication of the activation requirements will be probed by simulating ligand receptor interactions with 3D models of the opioid receptors constructed in our laboratory. Mutation of residues identified from these studies to be important in activation will be carried out and their pharmacological effects determined. Use of the 3D models for explicit characterization of ligand receptor complexes will bridge the gap between structure and function that cannot be filled by studies of the ligands alone. Focusing on the mouse and its three cloned receptors will maximize the consistency of the results obtained and the mechanistic and therapeutic inferences draw from them.