TRPA1 and General Anesthetics

描述（由申请人提供）：全身麻醉剂（GAs）是一组不同的化学物质，具有诱导可逆无意识的共同能力。尽管这些药物已经彻底改变了外科手术，但它们仍然不够理想。特别是，许多这些药物在服用时引起疼痛/刺激。吸入性气体可刺激气道和激活交感神经。此外，越来越多的证据表明，有毒麻醉剂有可能加剧术后疼痛和炎症。全世界每年大约进行2亿例手术，因此，了解GAs如何与TRP通道相互作用是非常必要的。我们之前的数据表明，地氟醚/异氟醚通过选择性激活“芥油”受体（TRPA1）来刺激感觉神经。本提案的主要目标是阐明TRPA1中对麻醉激动作用至关重要的氨基酸。我们假设地氟醚与TRPA1的第5和第6跨膜结构域之间的空腔结合。我们将探讨两个具体目标。(1)利用遗传学方法，我们将确定地氟醚激活TRPA1所需的假定位点。我们的初步数据揭示了TRPA1跨膜5在麻醉感知中的关键作用，因此我们将重点研究该区域的氨基酸。此外，我们将建立地氟醚与TRPA1对接的模型，以验证我们的基因/功能数据，并验证地氟醚直接与TRPA1相互作用。(2)我们将测试地氟烷类似物（包括对映体）激活TRPA1的能力。这些类似物旨在探测地氟醚中预测与TRPA1特异性相互作用的甲氧基的空间和电子性质。

DESCRIPTION (provided by applicant): General anesthetics (GAs) are a diverse group of chemicals with the shared ability to induce reversible unconsciousness. Although these drugs have revolutionized surgery they are still less than ideal. In particular, many of these drugs provoke pain/irritation upon administration. Inhalant GAs can provoke airway irritation and sympathetic activation. Moreover, there is emerging evidence that noxious anesthetics have the potential to exacerbate post-surgical pain and inflammation. Approximately 200 million surgeries are performed worldwide each year, and therefore, understanding how GAs interacts with TRP channels is highly desirable. Our previous data show that desflurane/isoflurane excites sensory nerves by selectively activating the "mustard-oil" receptor (TRPA1). A major goal of this proposal is to elucidate amino acids in TRPA1 that are critical for anesthetic agonism. We hypothesize that desflurane binds to a cavity between the 5th and 6th transmembrane domains of TRPA1. We will explore 2 specific aims. (1) Using a genetic approach we will determine the putative sites on TRPA1 necessary for activation by desflurane. Our preliminary data reveal a critical role for transmembrane 5 of TRPA1 in anesthetic sensing, and thus we will focus on amino acids in this region. Further, we will model docking of desflurane to TRPA1 to validate our genetic/functional data and verify that desflurane directly interacts with TRPA1. (2) We will test the ability of desflurane analogs (including enantiomers) to activate TRPA1. These analogs are designed to probe the steric and electronic properties of the methoxy group in desflurane predicted to specifically interact with TRPA1.