Diterpines as Selective Kappa Opioid Receptor Agonists

描述（由申请人提供）：Salvinorin A -致幻剂鼠尾草的活性成分-与鼠尾草一起代表了美国和其他地方滥用的新兴药物。Salvinorin A和鼠尾草提取物在人类中引起强烈而短暂的幻觉体验，这与经典致幻剂LSD、裸盖菇素和美斯卡灵不同。2002年，我的实验室发现salvinorin A能有效地特异性靶向？-阿片受体（KOR） （Roth et al., PNAS 2002）；这些发现已被广泛复制。尽管对salvinorin A的作用有浓厚的兴趣，但我们仍然不完全了解其在KOR中具有出色效力和选择性的分子和原子机制。这些研究的目的是发现salvinorin A如何结合并激活KOR。为了实现这些目标，我们将进行两个具体的目标：(1)阐明？-阿片受体对salvinorin A的作用至关重要，使用高通量分子生物学和(2)阐明？-阿片受体通过直接生化途径负责salvinorin A的作用。这些研究将阐明这种滥用药物是如何在原子水平上发挥作用的。这些研究具有以下重要意义：概念性：Salvinorin A是一种新兴的滥用药物，对人类的感知有深远的影响。Salvinorin A通过有效和选择性激活？阿片受体。了解salvinorin A如何实现其对KORs的卓越选择性和有效性是阐明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亲和和不可逆的salvinorin A类似物来鉴定KOR上负责结合的残基。据我们所知，这将是阿片受体领域首次对结合袋中的残基进行生物化学鉴定的研究。

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.