TRPV Pharmacophores from Cnidaria Venom

TRPV阳离子通道被认为在伤害性感受和神经源性炎症的诱导中起作用。 我们假设西尼达林后的剧烈疼痛和一系列神经炎症效应 人类的毒液是由TRPV通道传感器介导的，蛇毒含有新的 在TRPV通道上处于活动状态的组件。最近，我们已经证实了触手中的trpv1活性。 蛇床子属各主要类别的提取物。自从TRPV阳离子通道被认为在 伤害性感受和神经源性炎症的诱导，我们假设西尼达林的某些作用 毒液是由TRPV通道感受器介导的，蛇毒含有新的神经活性 以及在TRPV上活跃的免疫活性药效团。支持一个角色的另一个令人信服的论点 在蛇毒中的TRPV是已知的TRPVs配体之间显著的化学保守性 和蛇毒的成分。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)，寻求识别和表征新的 靶向毒蛇毒液中TRPV阳离子通道的药效团。我们的 实验计划采用生物测定指导的分离方法，以及组合光谱方法 用于检测、纯化和鉴定新发现的TRPV1药效团。 具体目的1.筛选翅毒中的TRPV活性成分。 低至中通量筛选协议，集成了新的生化和常规 电生理学TRPV法，将被用来鉴定三角帆蛇蛇毒中的TRPV1激动剂。 特定目的2.分离纯化并鉴定罗非鱼毒液中的TRPV药效团。 这些早期的后生动物TRPV药效团将通过成对的生化提纯/生物测定来分离 技术，并用高效液相色谱(HPLC)、气相色谱(GC)、 以及质谱学(MS)和核磁共振(核磁共振)谱。 我们专注于自然产物发现系统，最大限度地降低了这类天然产品发现工作中的固有风险。 也就是说，从这些古老的后生动物中寻找具有TRPV1活性的新结构的结果将不会 简单概括一下以前报道的生物活性化合物。这项工作的双重影响将是提供 迫切需要的，TRPV渠道的新药理学，并获得对病理的机械性见解 蛇毒的影响。鉴于最近显示出显著的治疗潜力的进展 TRPV1激动剂以及拮抗剂在治疗与慢性炎症和 在外科领域，来自蛇床子的新型TRPV1药物载体的潜在治疗效用非常高。TRPV蛋白是开发新止痛药的靶点。夏威夷盒子果冻的毒液 FISH含有潜在的针对TRPV通道的新型化合物。我们将探索化学多样性 以发现用于治疗神经疾病的新的先导化合物。

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.