Novel Analgesics from Australian Funnel-Web Spider Venom

来自澳大利亚漏斗网蜘蛛毒液的新型镇痛药

• 批准号: 7844819
• 负责人: Michael Nitabach
• 金额: $ 20.69万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-05-15 至 2012-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7844819
• 关键词: Acids; Adaptor Signaling Protein; Affinity; Analgesics; Area; Axon; Binding; Cell membrane; Chemicals; Complementary DNA; Complex; Conscious; Development; Drosophila genus; Economics; Family; Fiber; Future; Gated Ion Channel; Genomics; Goals; HTR3A gene; Heating; Home environment; Homologous Gene; Human; Inflammation Mediators; Inflammatory; Injury; Insecta; Institution; Internet; Ion Channel; Laboratories; Lead; Libraries; Malignant Neoplasms; Mass Spectrum Analysis; Mechanics; Messenger RNA; Modeling; Modification; National Institute of Neurological Disorders and Stroke; Neuraxis; Neurons; Neuropathy; Nociceptors; Oocytes; P2X-receptor; Pain; Peptides; Perception; Peripheral; Pharmaceutical Preparations; Pharmacologic Substance; Phosphorylation; Postoperative Pain; Proteomics; RNA Splicing; Research; Screening procedure; Sensory; Sensory Ganglia; Serotonin; Signal Transduction; Site; Source; Spiders; Stimulus; System; Testing; Toxin; Translational Research; Variant; Venoms; Vertebrates; Work; Xenopus laevis; Xenopus oocyte; combinatorial; in vivo; member; novel; pre-clinical; prevent; receptor; response; reversed phase chromatography; spider toxin FTX; transmission process; voltage

Pain caused by activation of pain-sensing peripheral neurons ("nociceptors") is a major source of human suffering and economic loss. Activation of nociceptors and the transmission of pain signals to the central nervous system, where they give rise to the conscious perception of pain, requires the coordinated participation of a variety of stimulus- and voltage-gated ion channels. Our long-term goal is the development of pharmaceutical agents for interfering with the functions of ion channels in mammalian nociceptors to ameliorate human pain. We will employ as a source of candidate agents a newly-discovered naturally-occurring combinatorial library of diverse peptide ion channel toxins ("atracotoxins") derived from the venom of Australian funnel-web spiders. To identify members of this library with desirable activity, we will screen them for activity against ion channels known to be important for nociceptor function. The first step in the transduction of painful stimuli-such as extreme heat or cold, noxious chemicals, mechanical injury, or inflammatory mediators-is the activation of stimulus-gated ion channels in the plasma membrane of nociceptors. Once a painful stimulus is transduced by stimulus-gated ion channels into an electrochemical signal, it must be transmitted to the CNS as an electrical signal down the axon of the nociceptor. The transmission of pain signals to the CNS requires activation of voltage-gated ion channels in the endings and axons of nociceptors. We will identify blockers of stimulus- and voltage-gated ion channels in the peripheral terminals of nociceptors by screening atracotoxins against cloned nociceptor ion channels expressed in Xenopus laevis oocytes. Once we have identified a subset of atracotoxins that are active against cloned nociceptor ion channels, we will test each of them against the corresponding native channels in acutely cultured sensory ganglion nociceptors. This secondary screen is essential for excluding from future in vivo studies of candidate analgesics those atracotoxins that are inactive against ion channels in their native state in the nociceptor plasma membrane. These studies will identify lead pharmaceutical agents for use as novel treatments for severe human pain with significant advantages over those that are currently available. In particular, by targeting ion channels present in the peripheral terminals of nociceptors, we open up the possibility for local drug application to the site of peripheral pain. Once these lead agents are identified, we will collaborate with established experts to test these agents in accepted in vivo preclinical mammalian models of pain.

由疼痛感测外周神经元（“伤害感受器”）的激活引起的疼痛是人类痛苦和经济损失的主要来源。伤害感受器的激活和疼痛信号向中枢神经系统的传递，在中枢神经系统中它们引起对疼痛的有意识感知，需要各种刺激门控和电压门控离子通道的协调参与。我们的长期目标是开发用于干扰哺乳动物伤害感受器中的离子通道功能以改善人类疼痛的药剂。我们将采用一个新发现的天然存在的组合库的不同肽离子通道毒素（“peptide ion channel toxins”），来自澳大利亚漏斗网蜘蛛的毒液作为候选药物的来源。为了鉴定该文库中具有所需活性的成员，我们将筛选它们对已知对伤害感受器功能重要的离子通道的活性。疼痛刺激（如极热或极冷、有毒化学物质、机械损伤或炎症介质）传导的第一步是激活伤害感受器质膜上的刺激门控离子通道。一旦疼痛刺激被刺激门控离子通道转换成电化学信号，它必须作为电信号沿着伤害感受器的轴突传递到CNS。疼痛信号传递到中枢神经系统需要激活伤害感受器末端和轴突中的电压门控离子通道。我们将通过筛选非洲爪蟾卵母细胞中表达的克隆伤害感受器离子通道的肉毒毒素来确定伤害感受器外周末端刺激和电压门控离子通道的阻断剂。一旦我们确定了一个亚组的肉毒毒素，对克隆的伤害感受器离子通道的活性，我们将测试他们对相应的天然通道，在急性培养的感觉神经节伤害感受器。这种二次筛选对于从候选镇痛剂的未来体内研究中排除那些在伤害感受器质膜中的天然状态下对离子通道无活性的肉毒毒素是必不可少的。这些研究将确定用于治疗严重人类疼痛的新型药物，这些药物比目前可用的药物具有显着优势。特别是，通过靶向离子通道存在于外周末梢的伤害感受器，我们打开了局部药物应用于外周疼痛的网站的可能性。一旦确定了这些先导药物，我们将与知名专家合作，在公认的临床前哺乳动物疼痛体内模型中测试这些药物。