Basic and Translational Research on General Anesthetics

项目摘要。全身麻醉，对潜在神经功能的可逆性药物抑制 意识、意识、记忆和伤害性运动反应是一项基本的医学干预措施。 显然需要具有可预测的动力学和降低毒性的新麻醉药，这将 促进医疗程序创新、疗效、效率和可及性。我为这些目标做出了贡献 使用两个严格和互补的战略。首先，我有严谨的基础科学知识 已建立的靶点，如GABAA受体中的分子麻醉机制。这项研究已经 发现不同全麻化学类型(依托咪酯)之间出人意料的特定相互作用 衍生物、甲基苯基烯丙基巴比妥酸盐或MPAB、神经类固醇，如α-黄酮类和苯甲酰醇) 以及典型突触GABAA受体中不同的跨膜间亚单位位点。我还介绍了 Monod-Wyman-Changeux(MWC)双状态共激动剂模型用于麻醉效应的定量分析 这些受体。其次，认识到针对GABAA受体的药物已被证明是唯一不令人满意的 临床全身麻醉剂，我是最早采用斑马鱼作为无偏药效学的人之一 发现可能通过多个分子靶点作用的新催眠化学类型的模型，并开发 加快机制研究的转基因品系。这款R35(MIRA)的广泛长期目标 未来五年及以后的拨款将进一步促进我们对麻醉剂的理解 分子水平上的机制，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-受体相互作用在 麻醉效果。这个项目将涉及的分子知识领域包括，但不是 仅限于提高GABAA受体麻醉部位定位的精确度，开发MWC模型 不同部位选择性麻醉剂对GABA调节作用的差异解释 化学类型，探测其他重要的(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型，以及 将这些结构功能途径扩展到其他麻醉剂敏感的五聚体配基门控离子 频道。我还将应用斑马鱼幼体和实时视频相结合的平台分析高达96 在药物库中识别新的镇静-催眠化学类型，并评估各种 镇静剂和催眠药的相互作用。新的催眠化学类型将在一组 使用电生理学和药理学工具的分子麻醉药靶点。我还将开发新的 麻醉剂靶蛋白基因敲除或敲入突变的转基因斑马鱼模型 测试这些靶点是否调节已有或已发现的镇静催眠药的行为效果。

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.