SOURCES OF NEUROACTIVE ANALGESIC N-ACYLETHANOLAMINES

性状（申请人摘要）：N-酰基乙醇胺（NAE）是有效的， 在哺乳动物中具有不同生理作用的生物活性脂质介质 包括但不限于疼痛起始的控制。Anandamide，a 以花生四烯酸为脂肪酸成分的NAE类型，是一种 哺乳动物脑中大麻素受体（CBI型）的内源性配体， 和短暂的细胞外积累的花生四烯酸酰胺（和其他NAE）， 参与神经元细胞中的“内源性大麻素”信号通路的一部分， 对疼痛的感知最近发现NAE富含 高等植物，我们建议在这种探索性的研究和发展， 阶段，以确定丰富的，天然来源的NAE混合物（结合 其它内源性大麻素脂质介质如酰基甘油）， 用于开发“天然止痛药”。“为了支持这一点 应用，准确的定量程序（气相色谱法与 质谱法），以确定这些脂质代谢产物的原油 植物提取物和工业精炼植物油馏分中。而且 采用快速筛选方法，可对各种提取物进行检测， 它们的神经活性。这种方法依赖于定量 动作电位模式的多通道电生理学测量 从微电极阵列（MEA）上培养的神经网络记录。的 几种内源性大麻素（包括一种种子） 混合NAE种类的提取物）对自发和荷包牡丹碱刺激的神经元的 网络活动有助于验证该系统的有效性。 CBI受体依赖性和非依赖性机制将通过 经典药理学方法（激动剂、拮抗剂、抑制剂等） 以及通过分子遗传学方法（来自CBI敲除的培养物 小鼠）。这项研究的结果应该针对新的植物来源， 开发神经活性治疗剂。

DESCRIPTION (APPLICANT'S ABSTRACT): N-Acylethanolamines (NAEs) are potent, bioactive lipid mediators with diverse physiological roles in mammals including, but not restricted to, the control of pain initiation. Anandamide, a type of NAE with arachidonic acid as the fatty acid constituent, is an endogenous ligand for the cannabinoid receptor (CBI type) in mammalian brain, and the transient extracellular accumulation of anandamide (and other NAEs) is part of the "endocannabinoid" signaling pathway in neuronal cells involved in the perception of pain. NAEs recently were found to be enriched in seeds of higher plants, and we propose in this exploratory research and development phase to identify abundant, natural sources of NAE-mixtures (combined with other endocannabinoid lipid mediators such as acylglycerols) which could be used in the development of "natural analgesics." In support of this application, accurate quantitative procedures (gas chromatography coupled with mass spectrometry) were developed to identify these lipid metabolites in crude plant extracts and in industrially-refined vegetable oil fractions. Moreover, a rapid screening method is in place by which various extracts can be tested for their neurological activities in vitro. This method relies on quantitative multi-channel electrophysiological measurements of action potential patterns recorded from neural networks cultured on microelectrode arrays (MEAs). The reproducible "quieting" effects of several endocannabinoids (including a seed extract of mixed NAE species) on spontaneous and bicuculline-stimulated neural network activity have helped to verify the efficacy of this system. CBI-receptor-dependent and -independent mechanisms will be identified by classical pharmacological approaches (agonists, antagonists, inhibitors, etc.) as well as by molecular genetic approaches (cultures derived from CBI-knockout mice). Results from this research should target new botanical sources for development as neuroactive therapeutics.