Discovery of analgesic diterpenoid alkaloids from medicinal Aconitum plants using a metabolomic approach

项目摘要 阿片类药物危机已经表明，对不针对阿片类药物的镇痛药物存在巨大需求。 阿片受体植物是开发这些药物的理想来源，因为它们产生各种各样的药物。 具有镇痛活性的天然产物，如水杨酸、辣椒素和大麻二酚。这些化合物作用于 动物的不同疼痛反应系统，比如内源性大麻素系统。一种植物属， 毛茛科，是世界各地不同传统药用系统中常用的植物，常以其止痛 和/或抗炎特性。在传统的中国医学中，冬虫夏草被用于 其生物活性主要与其二萜生物碱含量有关。而 这些化合物已被证明是有效的镇痛剂，有些还具有心脏毒性。尽管他们的相似 药用，美国的仙人掌物种还没有在化学方面进行彻底的研究， 生物活性我们研究的长期目标是识别、表征和量化治疗成分 作为新药来源的传统医药系统中使用的植物。该项目的总体目标是 分析北美的甘草，特别是关于它的镇痛二萜生物碱， 心脏毒性相比，更好地研究亚洲物种。我们最初的研究表明， 选择的二萜生物碱的化学结构和数量的变化，我们已经确定了fuziline 和来自亚洲红毛忍冬的新碱，它们具有镇痛活性而没有心脏毒性。其他 研究发现这些二萜生物碱靶向大麻素受体。因此，我们假设， 在美洲藜属物种中的一小组二萜生物碱将具有镇痛特性， 伴随心脏毒性。我们将通过实现三个目标来检验这一假设。在目标1中，我们将收集和 使用非靶向LC-QToF-MS检查八种蒲公英并确定二萜生物碱的模式 代谢组学研究。在目标2中，我们将评估从植物中提取的植物提取物、级分和纯化的二萜生物碱。 在高通量斑马鱼试验中鉴定靶向疼痛的生物活性化合物 但没有心脏毒性。在目标3中，我们将使用靶向LC-TQD-MS分析来定量靶向 镇痛化合物的八个物种，并加入代谢组学和生物学数据， 目的1和2一起通过新的代谢组学平台，如NP分析，以创建一个分子网络 的镇痛化合物产生的镇痛化合物，没有心脏毒性在一起，完成这三个 AIMS将产生一组镇痛化合物，这些化合物将优先用于进一步的临床前研究。这 该项目还将培训来自现代教育中代表性不足群体的本科生和研究生。 天然产物化学的应用和技术，并为他们提供就业机会 通过在会议上发言和撰写手稿来发展。

Project Summary The opioid crisis has made it apparent that there is a tremendous need for analgesic drugs that do not target the opioid receptors. Plants are an ideal source for developing these drugs, as they produce a diverse range of natural products with analgesic activity, like salicylic acid, capsaicin, and cannabidiol. These compounds act on different pain response systems in animals, like the endocannabinoid system. Aconitum, a genus of plants in the Ranunculaceae family, is used in different traditional medicine systems around the world, often for its analgesic and/or anti-inflammatory properties. In traditional Chinese medicine, Aconitum carmichaelii has been used for thousands of years, and the bioactivity has been associated mainly with its diterpenoid alkaloid content. While these compounds have been shown to be potent analgesics, some are cardiotoxic as well. Despite their similar medicinal use, American Aconitum species have not been as thoroughly studied in terms of chemistry or bioactivity. The long-term goal of our research is to identify, characterize, and quantify therapeutic components in plants used in traditional medicine systems as sources of new drugs. The overall objective of this project is to analyze Aconitum in North America, specifically regarding its analgesic diterpenoid alkaloids that lack cardiotoxicity as compared to the better-studied Asian species. Our initial studies on Aconitum showed significant variation in chemical structures and amounts of selected diterpenoid alkaloids, and we have identified fuziline and neoline from Asian Aconitum carmichaelii as having analgesic activity without being cardiotoxic. Other studies have found that these diterpenoid alkaloids target the cannabinoid receptors. Thus, we hypothesize that a small group of diterpenoid alkaloids in American Aconitum species will have analgesic properties without the concomitant cardiotoxicity. We will test this hypothesis by carrying out three aims. In Aim 1, we will collect and examine eight Aconitum species and determine patterns of diterpenoid alkaloids using untargeted LC-QToF-MS metabolomic studies. In Aim 2, we will assess plant extracts, fractions, and purified diterpenoid alkaloids from American Aconitum species in high-throughput zebrafish assays to identify bioactive compounds that target pain but do not have cardiotoxicity. In Aim 3, we will use a targeted LC-TQD-MS analysis to quantify the target analgesic compounds in the eight Aconitum species, and join the metabolomic and biological data obtained in Aims 1 and 2 together through novel metabolomic platforms, such as NP analyst, to create a molecular network of analgesic compounds produced by Aconitum that lack cardiotoxicity Together, the completion of these three aims will result in a group of analgesic compounds that will be prioritized for further preclinical studies. This project will also train undergraduate and graduate students from underrepresented groups in modern applications and technologies of natural product chemistry and provide them with opportunities for career development by presenting at conferences and authoring manuscripts.