Basic and Translational Research on General Anesthetics

项目摘要。全身麻醉，可逆的药理学抑制神经功能的基础 意识、意识、记忆和伤害性运动反应是必要的医学干预。 显然需要具有可预测的动力学和降低的毒性的新麻醉药物，其将 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，我有严格先进的基础科学知识 分子麻醉机制在建立的目标，如GABAA受体。本研究 揭示了不同的全身麻醉化学类型（依托咪酯）之间意外的特异性相互作用 衍生物、甲基-苯基烯丙基巴比妥酸盐或MPAB、神经甾体如阿法沙龙和苯甲酰基醇） 和典型的突触GABAA受体中不同的跨膜亚基间位点。我也介绍 Monod-Wyman-Changeux（MWC）双状态共激动剂模型用于定量分析麻醉剂对大鼠的作用 这些受体。第二，认识到GABAA受体特异性药物已被证明不能作为唯一的 临床全身麻醉剂，我是第一个采用斑马鱼作为一个公正的药效学 模型，以发现新的催眠化学型，可能通过多个分子靶点发挥作用，并开发 转基因株系，以加速机理研究。R35（MIRA）的长期目标 在未来五年及以后的时间里，我们将进一步推进我们对麻醉剂的理解。 在分子水平上的机制，发现新的化学家族与镇静催眠活性， 并创造新的转基因斑马鱼，以测试特定药物-受体相互作用在 麻醉效果本项目将涉及的分子知识领域包括但不限于 局限于提高GABAA受体麻醉部位定位的精度，开发MWC模型， 解释了不同部位选择性麻醉剂的不同激动剂与GABA调制效应 化学型，探测其他重要（如突触外）GABAA的亚基排列和结构 受体同种型，使用亚单位特异性麻醉剂和选择性影响其作用的突变，和 将这些结构-功能方法扩展到其他麻醉敏感的五聚体配体门控离子 渠道我还将应用平台结合斑马鱼幼虫和实时视频分析， 动物在同一时间，以确定新的镇静催眠药化学型的药物库，并评估各种 镇静催眠药物的相互作用新的催眠化学型将在一组 使用电生理学和药理学工具的分子麻醉剂靶点。我还将开发新的 在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 测试这些目标是否介导了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.