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.

激活疼痛感应周围神经元（“伤害感受器”）引起的疼痛是人类苦难和经济损失的主要来源。伤害感受器的激活以及将疼痛信号传播到中枢神经系统，在此引起了对疼痛的有意识的感知，就需要对各种刺激和电压门控离子通道进行协调的参与。我们的长期目标是开发用于干扰哺乳动物伤害感受器的离子通道功能以减轻人类疼痛的功能的药物。我们将用作候选代理的来源，新发现的自然存在的肽离子通道毒素（“阿抗毒素”）的自然存在组合图书馆，该库是从澳大利亚漏斗 - Web-Web蜘蛛的毒液中得出的。为了确定具有理想活动的该库的成员，我们将针对已知对伤害感受器功能很重要的离子通道进行筛查。疼痛刺激的转导的第一步，例如极高或冷热，有害化学物质，机械损伤或炎症介质 - 是伤害感受器质膜中刺激门控离子通道的激活。一旦通过刺激门控离子通道转导疼痛的刺激到电化学信号中，就必须将其传输到中枢神经系统，作为沿伤害感受器轴突的电信号。疼痛信号向中枢神经系统的传播需要激活伤害感受器的结尾和轴突中的电压门控通道。我们将通过筛选在Xenopus laevis卵母细胞中表达的克隆的伤害感受器离子通道来鉴定伤害感受器外周末端的刺激和电压门控离子通道的阻滞剂。一旦我们确定了针对克隆的伤害感受器离子通道活性的亚抗毒素的子集，我们将对急性培养的感觉神经节伤害感受器中的相应天然通道进行测试。该次要筛选对于将未来的候选镇痛药研究排除在外，这些镇痛药的体内研究是那些无活性的，这些毒素与Nocteptor质膜中的天然状态无活性。这些研究将确定铅药物用作严重人类疼痛的新型治疗方法，其优势与当前可用的那些疗法相比。特别是，通过靶向伤害感受器外围末端中存在的离子通道，我们为外周疼痛部位开放了局部药物的可能性。一旦确定了这些主要代理，我们将与已建立的专家合作，以在接受体内临床前哺乳动物模型的疼痛模型中测试这些代理。