Basic and Translational Research on General Anesthetics

项目摘要。全身麻醉，神经功能的可逆药理抑制 意识、意识、记忆和伤害性运动反应，是一项重要的医疗干预措施。 显然需要具有可预测动力学和降低毒性的新型麻醉药物，这将 促进医疗程序创新、功效、效率和可及性。我为这些目标做出了贡献 使用两种严格且互补的策略。首先，我拥有严格先进的基础科学知识 确定目标（如 GABAA 受体）的分子麻醉机制。这项研究有 揭示了不同全身麻醉化学类型（依托咪酯）之间出乎意料的特异性相互作用 衍生物、甲基苯基烯丙基巴比妥酸盐或 MPAB、神经类固醇如 alphaxalone 和苯甲酰醇） 以及典型突触 GABAA 受体中不同的跨膜亚基间位点组。我还介绍过 Monod-Wyman-Changeux (MWC) 二态协同激动剂模型，用于定量分析麻醉效果 这些受体。其次，认识到 GABAA 受体特异性药物作为单一药物已被证明不能令人满意。 临床全身麻醉剂，我是最早采用斑马鱼作为无偏见药效学的人之一 模型来发现可能通过多个分子靶标起作用的新催眠化学型，并开发 转基因系加速机制研究。 R35 (MIRA) 的广泛长期目标 未来五年及以后的资助将进一步增进我们对麻醉的理解 分子水平的机制，发现具有镇静催眠活性的新化学家族， 并创造新的转基因斑马鱼来测试特定药物受体相互作用的作用 麻醉作用。该项目将涉及的分子知识领域包括但不包括 仅限于提高 GABAA 受体麻醉位点定位的精度，开发 MWC 模型 解释不同位点选择性麻醉剂的不同激动剂与 GABA 调节效应 化学型，探测其他重要（例如突触外）GABAA 的亚基排列和结构 使用亚位点特异性麻醉剂和选择性影响其作用的突变的受体同种型，以及 将这些结构功能方法扩展到其他麻醉敏感的五聚配体门控离子 渠道。我还将应用该平台结合斑马鱼幼虫和多达96个的实时视频分析 一次对动物进行识别，以识别药物库中新的镇静催眠化学类型并评估各种 镇静催眠药物的相互作用。新的催眠化学类型将在一组中表征其效果 使用电生理学和药理学工具的分子麻醉目标。我也会开发新的 麻醉靶蛋白中具有敲除或敲入突变的转基因斑马鱼系作为模型 测试这些目标是否介导已建立或发现的镇静催眠药的行为效应。

Project Summary. General anesthesia, the reversible pharmacologic inhibition of neural functions underlying consciousness, awareness, memory, and nociceptive motor responses, is an essential medical intervention. There is a clear need for new anesthetic drugs with predictable kinetics and reduced toxicity, which will facilitate medical procedural innovation, efficacy, efficiency, and accessibility. I have contributed to these goals using two rigorous and complementary strategies. First, I have rigorously advanced basic science knowledge of molecular anesthetic mechanisms in established targets such as GABAA receptors. This research has revealed unexpectedly specific interactions between different general anesthetic chemotypes (etomidate derivatives, methyl-phenyl allyl barbiturates or MPABs, neurosteroids like alphaxalone, and benzoyl alcohols) and distinct sets of transmembrane inter-subunit sites in typical synaptic GABAA receptors. I also introduced Monod-Wyman-Changeux (MWC) two-state co-agonist models for quantitative analysis of anesthetic effects in these receptors. Second, recognizing that GABAA receptor-specific drugs have proven unsatisfactory as sole clinical general anesthetic agents, I am among the first to adopt zebrafish as an unbiased pharmacodynamic model to discover new hypnotic chemotypes that may act via multiple molecular targets, and to develop transgenic lines to accelerate mechanisms research. The broad long-term objectives of this R35 (MIRA) grant during the next five years and beyond are to further advance our understanding of anesthetic mechanisms at the molecular level, to discover new chemical families with sedative-hypnotic activity, and to create new transgenic zebrafish to test the roles of specific drug-receptor interactions in anesthetic effects. Molecular knowledge areas that will be addressed by this project include, but are not limited to improving the precision of anesthetic site mapping in GABAA receptors, developing MWC models that account for differential agonist versus GABA modulation effects of the distinct site-selective anesthetic chemotypes, probing the subunit arrangement and structures of other important (e.g. extra-synaptic) GABAA receptor isotypes using subsite-specific anesthetics and mutations that selectively affect their actions, and extending these structure-function approaches to other anesthetic-sensitive pentameric ligand-gated ion channels. I will also apply the platform combining zebrafish larvae and real-time video analysis of up to 96 animals at a time to identify new sedative-hypnotic chemotypes in drug libraries and assess a variety of sedative-hypnotic drug interactions. New hypnotic chemotypes will be characterized for effects in a panel of molecular anesthetic targets using electrophysiology and pharmacologic tools. I will also develop new transgenic zebrafish lines with knock-out or knock-in mutations in anesthetic target proteins as models for testing if these targets mediate the behavioral effects of established or discovered sedative-hypnotics.