Structure-based design, synthesis, and biochemical and pharmacological characterization of novel salvinorin A analogues as active state probes of the kappa-opioid receptor.
Structure-based design, synthesis, and biochemical and pharmacological characterization of novel salvinorin A analogues as active state probes of the kappa-opioid receptor.
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基于结构的设计，合成以及新型Salvinorin A类似物作为Kappa-阿片受体的活性态探针的生化和药理表征。
DOI：
10.1021/bi900605n
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发表时间：
2009-07-28
期刊：
影响因子：
2.9
通讯作者：
Roth, Bryan L.
中科院分区：
文献类型：
--
作者：
Yan, Feng;Bikbulatov, Ruslan V.;Mocanu, Viorel;Dicheva, Nedyalka;Parker, Carol E.;Wetsel, William C.;Mosier, Philip D.;Westkaemper, Richard B.;Allen, John A.;Zjawiony, Jordan K.;Roth, Bryan L.
鼠尾草素A（Salvinorin A）是最强的天然致幻剂，自从κ-阿片受体（k-opioid receptor，KOR）被确定为主要的作用靶点后，其作用越来越受到人们的关注。在这里，我们报告的设计，合成和生化表征的新的，不可逆的，鼠尾草素A衍生的配体适合作为活性状态的探针的KOR。基于先前的取代的半胱氨酸可及性和分子建模研究，选择C3157.38作为共价标记鼠尾草素A衍生配体的潜在锚定点。一系列潜在的共价结合配体的自动对接表明，卤代乙酸酯部分或其他类似的亲电基团可以不可逆地与C3157.38结合。22-发现硫氰酸根鼠尾草素A（RB-64）和22-氯鼠尾草素A（RB-48）在体外和体内都是非常有效和选择性的KOR激动剂。如基于分子建模研究所预测的，RB-64诱导结合的耐洗性抑制，其严格要求结合口袋中或附近的游离半胱氨酸。利用合成KOR肽和RB-64的质谱（MS）研究支持锚定残基为C3157.38的假设，并提出了共价结合的一种生化机制。这些研究提供了在KOR的激动剂结合状态下存在游离半胱氨酸的直接证据，并提供了对鼠尾草素A结合并激活KOR的机制的新见解。
Salvinorin A, the most potent naturally occurring hallucinogen, has gained increasing attention since the κ-opioid receptor (KOR) was identified as its principal molecular target by us. Here we report the design, synthesis and biochemical characterization of novel, irreversible, salvinorin A-derived ligands suitable as active state probes of the KOR. Based on prior substituted cysteine accessibility and molecular modeling studies, C3157.38 was chosen as a potential anchoring point for covalent labeling of salvinorin A-derived ligands. Automated docking of a series of potential covalently-bound ligands suggested that either a haloacetate moiety or other similar electrophilic groups could irreversibly bind with C3157.38. 22-thiocyanatosalvinorin A (RB-64) and 22-chlorosalvinorin A (RB-48) were both found to be extraordinarily potent and selective KOR agonists in vitro and in vivo. As predicted based on molecular modeling studies, RB-64 induced wash-resistant inhibition of binding with a strict requirement for a free cysteine in or near the binding pocket. Mass spectrometry (MS) studies utilizing synthetic KOR peptides and RB-64 supported the hypothesis that the anchoring residue was C3157.38 and suggested one biochemical mechanism for covalent binding. These studies provide direct evidence for the presence of a free cysteine in the agonist-bound state of KOR and provide novel insights into the mechanism by which salvinorin A binds to and activates KOR.
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影响因子：
56.9
作者：
Cohen, MS;Zhang, C;Taunton, J
通讯作者：
Taunton, J
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作者：
MANDA, S;LERNERMARMAROSH, N;ABOOD, LG
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16.8
作者：
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作者：
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