TRPV Pharmacophores from Cnidaria Venom

刺胞动物毒液 TRPV 药效团

基本信息

批准号：
7532828
负责人：
ANGEL ANNE YANAGIHARA
金额：
$ 24.06万
依托单位：
UNIVERSITY OF HAWAII AT MANOA
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-07-01 至 2010-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7532828
关键词：
Afferent Neurons Agonist Amines Anti Inflammatory Analgesics Biochemical Biological Assay Biological Availability Biological Factors Boxing Cannabinoids Carybdea Cations Chemical Structure Chemicals Chronic Class Clinical Cnidaria Cnidarian Venoms Complex Data Detection Development Eligibility Determination Evaluation Family Fishes Fourier Transform Fractionation Freezing Future G Protein-Coupled Receptor Genes Gas Chromatography Goals Hawaiian population High Pressure Liquid Chromatography Human Inflammation Inflammatory Inflammatory Response Lead Ligands Lipids Liquid substance Mass Chromatography Mass Spectrum Analysis Mediating Methodology Methods Molecular NMR Spectroscopy Neurogenic Inflammation Neurons Nitrogen Nociception Nuclear Magnetic Resonance Operative Surgical Procedures Organ Outcome Pain Peripheral Personal Satisfaction Pharmaceutical Preparations Pharmacology Property Proteins Reporting Research Research Project Grants Research Proposals Respiratory Failure Risk Role Screening procedure Sonication Source Spectrometry Sting Injury Structure System TRPV channel TRPV1 gene Techniques Testing Therapeutic Therapeutic Agents Therapeutic Studies Tissues Venoms Visceral Work aqueous base capsule cell type concept cross reactivity drug discovery evaporation human tissue infrared spectroscopy insight nervous system disorder novel pharmacophore research study response sensor small molecule

项目摘要

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.

TRPV 阳离子通道在伤害感受和神经源性炎症的诱导中发挥作用。我们假设刺胞动物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，并且刺胞动物的毒液含有新的 TRPV 通道上处于活动状态的组件。最近，我们在触手中证明了 TRPV1 活性刺胞动物所有主要类别的提取物。由于 TRPV 阳离子通道已被赋予以下作用：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毒液由 TRPV 通道传感器介导，刺胞动物的毒液含有新型神经活性物质以及在 TRPV 上活跃的免疫活性药效团。支持角色的另一个令人信服的论据对于刺胞动物毒液中的 TRPV 而言，TRPV 的已知配体之间具有显着的化学保守性和刺胞动物毒液的成分。这项修订后的探索性研究提案是为了回应 PAR-07-048（治疗神经系统疾病的药物发现），旨在识别和表征新颖的药物刺胞动物 Carybdea alata 毒液中靶向 TRPV 阳离子通道的药效团。我们的实验计划采用生物测定导向的分馏方法和组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. alata 毒液中的 TRPV 活性化合物。低到中通量筛选方案，整合了新型生化和常规筛选方案电生理 TRPV 测定将用于鉴定 C. alata 毒液中的 TRPV1 激动剂。具体目标 2. 从 C. alata 毒液中纯化和表征 TRPV 药效团。这些早期后生动物 TRPV 药效团将使用配对生化纯化/生物测定进行分离技术并通过高效液相色谱（HPLC）、气相色谱（GC）进行表征，和质谱（MS）和核磁共振（NMR）光谱。我们对刺胞动物系统的关注最大限度地降低了此类天然产品发现工作的固有风险。也就是说，探索这些古老后生动物具有 TRPV1 活性的新结构的结果不会简单概括一下之前报道的生物活性化合物。这项工作的双重影响将是提供 TRPV 通道急需的新型药理学，并获得病理机制的见解刺胞动物毒液的影响。鉴于最近的进展表明了显着的治疗潜力 TRPV1 激动剂以及拮抗剂在治疗与慢性炎症相关的疼痛和手术中，来自刺胞动物的新型 TRPV1 药效团的潜在治疗效用非常高。 TRPV 蛋白是开发新止痛药的目标。夏威夷果冻的毒液鱼含有针对 TRPV 通道的潜在新型化合物。我们将探索化学多样性利用这种毒液来发现治疗神经系统疾病的新先导化合物。