Project 3 - In vivo pharmacology of cannabinoid receptor probes

项目摘要 内源性大麻素系统的治疗潜力尚未转化为安全有效。 药物。翻译鸿沟，或“死亡之谷”，将有希望的临床前研究与 一种新批准的药物的鉴定。功能选择性(或有偏向的激动症)是最近受到重视的 CB1和CB2受体信号传导的特性。临床翻译面临的挑战包括 信号通路最能预测治疗效果。因此，有必要确定候选人 配体参与治疗所需的特定信号通路，而理想情况下， 绕过那些会产生不良副作用的途径。有偏向的激动症和变构 调制代表了利用内源性大麻素治疗潜力的替代策略 没有直接作用的CB1激动剂的有害影响的信号系统。信号通路 CB1和CB2治疗效果的必要和可有可无仍然知之甚少。架起这座桥梁 如果我们要避免临床翻译中代价高昂的失败，知识差距是至关重要的。项目3将使用新颖的CB1 和CB2探针，在项目1中开发并在项目2中体外表征，以阐明CB1如何 偏向激动剂、探针特异性CB1正性变构调节和CB2选择性激动剂对体内的影响 药理学。目的1定义具有功能选择性的CB1探针的体内药理学。我们将为您介绍 功能选择性CB1激动剂以及紧密结合(慢Koff)和探针特异性CB1 PAM(即 在我们的初步体内研究中显示出高效力)对于CB1激活的主要迹象，抑制能力 神经性伤害性感觉以及导致耐受和身体依赖的倾向。目标2将定义 CB2激动剂对CB2的选择性高于CB1的体内药理学。我们会确定 是否可以通过掺入高亲和力的CB2来改善体内药理特性和选择性 缺乏CB1激动症的激动症。我们还将确定体内药理学特征是否可以 通过将CB1拮抗剂(中性、反向激动剂、部分激动剂)与高亲和力CB2结合来增强 激励性。这一目的将比较表现出这些性质的配体的光谱以实现大麻仿效， 抗痛觉异常的功效和耐受性，以及诱发身体依赖和负性反应的能力 情感状态。该项目将验证改进的CB1和CB2探针，而不是开发药物。 阐明体内效应和副作用所必需和不可缺少的信号通路 将改进以大麻素为基础的疗法，并打破成功临床翻译的障碍。

Project Summary The therapeutic potential of the endocannabinoid system has yet to translate into safe and effective medicines. The translation gap, or “valley of death” separates promising preclinical research from identification of a newly approved drug. Functional selectivity (or biased agonism) is a recently appreciated property of CB1 and CB2 receptor signalling. Challenges for clinical translation include the question of what signalling pathway best predicts therapeutic efficacy. It is, therefore, necessary to establish that candidate ligands engage the specific signalling pathways necessary for therapeutic benefit, while, ideally, circumventing those pathways responsible for unwanted side effects. Both biased agonism and allosteric modulation represent alternative strategies to harness the therapeutic potential of the endocannabinoid signalling system without the unwanted effects of direct acting CB1 agonists. The signalling pathways necessary and dispensable for CB1 and CB2 therapeutic efficacy remain poorly understood. Bridging this gap in knowledge is critical if we are to avoid costly failures in clinical translation. Project 3 will use novel CB1 and CB2 probes that are developed in Project 1 and characterized in vitro in Project 2 to elucidate how CB1 biased agonism, probe specific CB1 positive allosteric modulation and CB2 selective agonism impact in vivo pharmacology. Aim 1 will define the in vivo pharmacology of functionally selective CB1 probes. We will profile functionally selective CB1 agonists as well as tight binding (slow Koff) and probe-specific CB1 PAMs (that exhibit high potency in our preliminary in vivo studies) for cardinal signs of CB1 activation, ability to suppress neuropathic nociception as well as propensity to induce tolerance and physical dependence. Aim 2 will define the in vivo pharmacology of CB2 agonists with improved selectivity for CB2 over CB1. We will ascertain whether in vivo pharmacological profiles and selectivity can be improved by incorporating high affinity CB2 agonism with a lack of CB1 agonism. We will also determine whether in vivo pharmacological profiles can be enhanced by incorporating CB1 antagonism (neutral, inverse agonism, partial agonism) with high affinity CB2 agonism. This Aim will compare a spectrum of ligands exhibiting these properties for cannabimimetic effects, anti-allodynic efficacy and tolerance, as well as capacity to induce physical dependence and negative affective states. This project will validate improved CB1 and CB2 probes, not to develop a medication. Elucidation of signalling pathways necessary and dispensable for in vivo efficacy and unwanted side effects will improve cannabinoid-based therapeutics and break down barriers to successful clinical translation.