Diterpines as Selective Kappa Opioid Receptor Agonists

描述(由申请者提供)：丹参甲素--致幻鼠尾草的有效成分--与丹参一起代表着美国和其他地方的新兴滥用药物。丹参甲素和丹参提取物会引起人类强烈而短暂的幻觉体验，这与经典的致幻剂LSD、裸盖菇素和梅斯卡林不同。2002年，我的实验室发现salvinorin A有效且特定地针对？-阿片受体(KOR)(Roth等人，PNAS 2002)；这些发现已被广泛复制。尽管Salvinorin A的作用引起了人们的极大兴趣，但我们仍然不完全了解在KOR中导致其精致效力和选择性的分子和原子机制。这些研究的目的是发现丹参甲素如何与KOR结合并激活KOR。为了实现这些目标，我们将进行两个具体的目标：(1)利用高通量分子生物学阐明对丹参素A的作用至关重要的β-阿片受体的结构特征；(2)通过直接生化方法阐明与丹参素A作用有关的β-阿片受体的结构特征。这些研究将阐明这种滥用药物如何在原子水平上发挥作用。这些研究的意义如下：概念：丹参甲素是一种新兴的滥用药物，它对人类的感知产生了深远的影响。丹参素A通过有效和选择性地激活β-阿片受体发挥这些作用。了解Salvinorin A如何实现其对Kors的显著选择性和效力是阐明Salvinorin A作用机制的重要第一步。技术：我们将利用我们发明的新颖且非常强大的基于酵母的技术(ArmBruster等人，2007a)来确定Salvinorin A与Kors结合并激活Kors的分子和原子机制。我们还将使用新合成的PM亲和力和不可逆萨尔维甲素A类似物来鉴定KOR上负责结合的残基。据我们所知，这将是在阿片受体领域进行的第一次生化鉴定结合口袋中残留物的研究。

DESCRIPTION (provided by applicant): Salvinorin A - the active ingredient of the hallucinogenic sage Salvia divinorum - along with Salvia divinorum represent emerging drugs of abuse in the US and elsewhere. Salvinorin A and extracts of Salvia divinorum induce an intense and short-lived hallucinatory experience in humans unlike that of the classical hallucinogens LSD, psilocybin and mescaline. In 2002, my lab discovered that salvinorin A potently and specifically targets ?-opioid receptors (KOR) (Roth et al., PNAS 2002); these findings have been widely replicated. Despite intensive interest in the actions of salvinorin A, we still do not fully understand the molecular and atomic mechanisms responsible for its exquisite potency and selectivity at KOR. The goal of these studies is to discover how salvinorin A binds to and activates KOR. To accomplish these goals we will conduct two specific aims: (1) elucidate the structural features of ?-opioid receptors essential for salvinorin A's actions using high-throughput molecular biology and (2) elucidate the structural features of ?-opioid receptors responsible for salvinorin A's actions via direct biochemical approaches. These studies will clarify how this drug of abuse exerts its actions at the atomic level. These studies are significant as follows: Conceptual: Salvinorin A is an emerging drug of abuse which has profound effects on human perception. Salvinorin A exerts these actions via potent and selective activation of ?-opioid receptors. Understanding how salvinorin A achieves its remarkable selectivity and potency for KORs is an essential first step toward elucidating the mechanism of action of salvinorin A. Technical: We will utilize novel and extraordinarily robust yeast-based technologies we have invented (Armbruster et al., 2007a) to identify the molecular and atomic mechanisms by which salvinorin A binds to and activates KORs. We will also employ newly synthesized pM-affinity and irreversible salvinorin A analogues to identify residues on KOR responsible for binding. To our knowledge, these will be the first studies in the opioid receptor field to biochemically identify residues in the binding pocket.