SOURCES OF NEUROACTIVE ANALGESIC N-ACYLETHANOLAMINES

说明(申请人摘要)：N-乙酰乙醇胺(NAE)是一种在哺乳动物中具有多种生理作用的有效的生物活性脂质介体 包括但不限于对疼痛起始的控制。阿南达胺，一种 以花生四烯酸为脂肪酸成分的NAE类型，是一种 哺乳动物大脑中大麻素受体(CBI型)的内源性配体， 而阿南达胺(和其他NAE)的瞬时胞外蓄积是 神经细胞中“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中含有丰富的 高等植物，我们建议在这一探索性的研究和开发 阶段，以确定NAE混合物的丰富自然来源(与 其他内源性大麻类脂质介体，如酰甘油)，可能是 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(气相色谱与 质谱仪)用于鉴定原油中的这些脂类代谢物 植物提取物和工业精炼植物油馏分中。此外，一个 建立了快速筛选方法，可对各种提取物进行检测 他们在体外的神经活动。这种方法依赖于定量的 动作电位模式的多通道电生理测量 记录自培养在微电极阵列(MEA)上的神经网络。这个 几种内源性大麻素(包括一粒种子)可重现的“镇静”作用 混合NAE提取物)对自发神经和荷包牡丹碱刺激的神经的影响 网络活动有助于验证这一系统的有效性。 CBI受体依赖和非依赖的机制将通过 经典药理方法(激动剂、拮抗剂、抑制剂等) 以及分子遗传学方法(来源于CBI-基因敲除的培养 老鼠)。这项研究的结果应该针对新的植物学来源 作为神经活性疗法的发展。

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.