TRPV Pharmacophores from Cnidaria Venom

TRPV阳离子通道被认为在伤害感受和神经源性炎症的诱导中起作用。 我们假设刺胞动物注射后的剧烈疼痛和神经炎症效应 人类中的毒液注入是由TRPV通道传感器介导的，并且刺胞动物毒液含有新的 在TRPV通道上活跃的组件。最近，我们已经证明了TRPV1活性在tennis 所有主要刺胞纲的提取物。由于TRPV阳离子通道已被赋予的作用， 伤害感受和神经源性炎症的诱导，我们假设刺胞动物的某些作用 注毒是由TRPV通道传感器介导的，刺胞动物毒液含有新的神经活性物质， 和对TRPV有活性的免疫活性药效团。另一个令人信服的论点是 TRPVs在刺胞动物毒液蛰入中的作用是TRPVs的已知配体之间的显著化学保守性 和刺胞动物毒液的成分。这份修订后的探索性研究提案是为了回应 PAR-07 - 048（神经系统疾病的药物发现），旨在鉴定和表征新的 靶向刺胞动物Carybdea阿拉塔毒液中TRPV阳离子通道的药效团。我们 实验计划采用生物测定指导的分馏方法，并结合光谱方法 用于新发现的TRPV1药效团的检测、纯化和表征。 具体目标1。筛选C.阿拉塔毒液。 低至中等通量筛选方案，整合了新的生物化学和传统的生物技术， 电生理学TRPV测定将用于鉴定C.阿拉塔毒液。 具体目标2。纯化并表征来自C.阿拉塔毒液。 这些早期后生动物TRPV药效团将使用成对的生化纯化/生物测定分离 技术并通过高效液相色谱法（HPLC），气相色谱法（GC）， 质谱（MS）和核磁共振（NMR）光谱。 我们对刺胞系统的关注最大限度地减少了这种天然产物发现工作的固有风险。 也就是说，从这些古老的后生动物中探索具有TRPV1活性的新结构的结果将不会 简单概括了以前报道生物活性化合物。这项工作的双重影响将是提供 迫切需要的，新的药理学TRPV通道，并获得机制的见解，病理 刺胞动物毒液的影响。鉴于最近的进展表明， TRPV1激动剂以及拮抗剂在治疗与慢性炎症相关的疼痛中的应用， 在外科手术中，来自刺胞动物的新型TRPV 1药效团的潜在治疗效用非常高。TRPV蛋白是开发新止痛药的靶点。夏威夷盒水母的毒液 鱼类含有潜在的靶向TRPV通道的新化合物。我们将探索化学多样性 研究这种毒液的目的是为了发现新的先导化合物来治疗神经系统疾病。

TRPV cation channels have been ascribed roles in nocioception and the induction of neurogenic inflammation. We hypothesized that the intense pain and constellation of neuro-inflammatory effects following cnidarian envenomation in humans are mediated by TRPV channel-sensors, and that cnidarian venoms contain novel components that are active on TRPV channels. Recently, we have demonstrated TRPV1 activity in tentacle extracts of all major classes of Cnidaria. Since TRPV cation channels have been ascribed roles in nocioception and the induction of neurogenic inflammation, we hypothesized that certain effects of cnidarian envenomation are mediated by TRPV channel-sensors and that cnidarian venoms contain novel neuroactive and immunoactive pharmacophores which are active at TRPVs. Another compelling argument in favor of a role for TRPVs in cnidarian envenomation is the marked chemical conservation between known ligands for TRPVs and components of cnidarian venoms. This revised exploratory research proposal, submitted in response to PAR-07-048 (Drug Discovery for Nervous System Disorders), seeks to identify and characterize novel pharmacophores that target TRPV cation channels in the venom of the cnidarian, Carybdea alata. Our experimental plan employs bioassay-directed fractionation methods, and combined spectroscopic approaches for the detection, purification and characterization of newfound TRPV1 pharmacophores. Specific Aim 1. Screen for TRPV-active compounds in C. alata venom. Low- to medium-throughput screening protocols, which integrate novel biochemical and conventional electrophysiological TRPV assays, will be employed to identify TRPV1 agonists in C. alata venom. Specific Aim 2. Purify and characterize TRPV pharmacophores from C. alata venom. These early metazoan TRPV pharmacophores will be isolated using paired biochemical purification/bioassay techniques and characterized by high-performance liquid chromatography (HPLC), gas chromatography (GC), and mass spectrometry (MS), and nuclear magnetic resonance (NMR) spectroscopy. Our focus on the cnidarian system minimizes the inherent risk in this type of natural product discovery effort. That is, the outcome from probing these ancient metazoans for novel structures with TRPV1 activity will not simply recapitulate previously reported bioactive compounds. The dual impact of this work will be to provide much-needed, novel pharmacology for TRPV channels, and to gain mechanistic insights into the pathological effects of cnidarian venoms. In view of recent advances which demonstrate the marked therapeutic potential of TRPV1 agonists, as well as antagonists, in the treatment of pain associated with chronic inflammation and surgery, the potential therapeutic utility of novel TRPV1 pharmacophores from cnidaria is extremely high. TRPV proteins are targets for the development of new pain medications. The venom of the Hawaiian box jelly fish contains potentially novel compounds which target TRPV channels. We will explore the chemical diversity of this venom to discover new lead compounds for the management of neurological disorders.