Protection against sarin-induced neurotoxicity via an in vivo caspase inhibitor

通过体内半胱天冬酶抑制剂防止沙林引起的神经毒性

• 批准号: 8019732
• 负责人: DAVID R COOL
• 金额: $ 33.82万
• 依托单位: WRIGHT STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-30 至 2012-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8019732
• 关键词: Acetylcholinesterase; Address; Animal Model; Anti-Inflammatory Agents; Anti-inflammatory; Anticonvulsants; Antidotes; Astrocytes; Atropine; Attenuated; Biological; Biological Markers; Brain; Brain Injuries; Caspase; Caspase Inhibitor; Cause of Death; Cell Death; Cells; Chemical Warfare; Chemical Warfare Agents; Chemicals; Confocal Microscopy; Convulsions; Cytokine Activation; Development; Diazepam; Disease; Drug Combinations; Drug Design; Encephalitis; Enzymes; Exposure to; Future; Goals; Health; Health Benefit; Hour; Image; Immune; Inflammation; Inflammatory; Inflammatory Response; Injection of therapeutic agent; Length; Mass Spectrum Analysis; Medical; Microglia; Military Personnel; Mus; Muscarinic Acetylcholine Receptor; Natural regeneration; Nerve Degeneration; Nervous System Trauma; Neurons; Neuropathy; Organophosphates; Outcome; Pathway interactions; Pharmaceutical Preparations; Population; Proteins; Regimen; Reporting; Research Project Grants; Risk; Sarin; Seizures; Societies; Stroke; Survivors; System; Technology; Testing; Time; Tissues; Toxic effect; Trauma; Treatment Protocols; base; brain cell; brain tissue; cell killing; cytokine; design; drug development; in vivo; in vivo Model; mouse model; nerve agent; neuroinflammation; neuron loss; neurotoxicity; novel; novel therapeutics; prevent; protective effect; quinoline-val-asp(OMe)-CH2-OPH; response; synthetic polymer Bioplex; toxic organophosphate insecticide exposure

DESCRIPTION (provided by applicant): Prolonged and progressive neuroinflammation and neurodegeneration are correlated with low-level exposure to chemical warfare agents (CWAs), such as sarin. Civilians surviving terrorist attacks with sarin are likely to be at most risk of these conditions due to the length of time required to administer treatment and the type of treatment currently available. Current US military treatment for CWA organophosphate exposure is a specialized drug combination containing; a drug to regenerate acetylcholinesterase activity; and two drugs to prevent seizures. This treatment must be administered within 40 minutes, after which there is no protection against seizures and progressive neurological damage occurs, including neuroinflammation. An understanding of the underlying causes of the neuroinflammatory response and ensuing cell death are currently lacking. Without this information, drug design and treatment are limited to similar types of drugs. The goal of this proposal is to test the ability of a novel broad-spectrum caspase inhibitor, Q-VD-OPh, to attenuate neuroinflammation and neurodegeneration in sarin treated mice when administered 30 minutes after exposure to sarin. Efficacy will be compared at 48 hours and 14 days with and without the anticonvulsant, diazepam and the negative cell death control, Q-VE-OPh. The first aim of the study will be to establish neuroinflammatory and cell death markers of sarin exposure in the mice. Aims two and three will be to determine if Q-VD-OPh can attenuate the progressive sarin-induced cell death and neuroinflammation. Since seizures and neurodegeneration are not mutually exclusive, in aim, 4, we will determine if Q-VD-OPh and diazepam together can be additive in preventing long-term sarin damage. The brain tissues will be analyzed for cytokine expression by multiplex (BioPlex) bead-based and by MALDI-Imaging mass spectrometry for localization of sarin and Q-VD-OPh. Confocal microscopy and immunostaining will be used to analyze tissue sections for specific caspases and other biomarkers of cell death. The medical and health benefits to society of this type of therapy may extend beyond a countermeasure for sarin-exposure for civilians to military applications as well as potentially for stroke, convulsive disorders and other neuropathies that result from trauma, chemical or biological insults.) PUBLIC HEALTH RELEVANCE: Terrorist attacks with sarin and other chemical warfare nerve agents are an ever present threat around the world. Civilian survivors of such attacks are at increased risk of problems associated with inflammation of the brain as well as brain cell death. Treatment of civilians will require development of drugs that can act within a half-hour to hours to days after sarin exposure. The research project outlined here is designed to test a novel drug (Q-VD-OPh) that can inhibit enzymes responsible for killing cells, i.e., caspases, and reduce inflammation by immune cells in the brain. Positive outcomes of the project would be less brain cell death and inflammation at both 48 hours and 14 days following sarin exposure. Further benefits will be the development of an array of inflammatory proteins and specific enzymes of cell death that can be used to drive the discovery of new drugs.

描述（由申请人提供）：长期和进行性神经炎症和神经变性与低水平接触化学战剂（CWAS）（例如Sarin）相关。由于执行治疗所需的时间和当前可用的治疗类型，对萨林遭受恐怖袭击的平民可能会处于这些状况的风险。美国目前对CWA有机磷酸盐暴露的军事疗法是一种专门的药物组合；再生乙酰胆碱酯酶活性的药物；和两种防止癫痫发作的药物。这种治疗必须在40分钟内给予这种治疗方法，之后没有防止癫痫发作的保护，并且发生了渐进的神经系统损害，包括神经炎症。目前缺乏对神经炎症反应和随之而来的细胞死亡的根本原因的理解。没有这些信息，药物设计和治疗仅限于类似类型的药物。该提案的目的是测试新型的广谱caspase抑制剂Q-VD-OPH的能力，在暴露于沙林后30分钟后，在Sarin处理的小鼠中衰减神经炎症和神经变性。有或没有抗惊厥药，地西epa和阴性细胞死亡控制，Q-VE-o-o-o-o-o-o-o-o-pe-o-o-o-o-48小时14天，将在48小时14天进行疗效。该研究的第一个目的是建立小鼠沙林暴露的神经炎症和细胞死亡标记。目的是两个和三个是确定Q-VD-OPH是否可以减弱进行性Sarin诱导的细胞死亡和神经炎症。由于癫痫发作和神经变性不是相互排斥的，因此，在AIM中，我们将确定Q-VD-o-o-o-ph和地西ep剂是否可以加在一起，以防止长期的沙林损害。将通过基于多路复用（Bioflex）珠子和MALDIMI-IMAGGING质谱法分析脑组织的细胞因子表达，以定位于Sarin和Q-VD-OPH。共聚焦显微镜和免疫染色将用于分析特定caspase和其他细胞死亡生物标志物的组织切片。这种类型的疗法对社会的医疗和健康益处可能超出了对平民对军事应用的侵害的对策，以及中风，抽搐疾病和其他因创伤，化学或生物学侮辱而导致的神经病的可能性。 公共卫生相关性：与沙林和其他化学战神经毒剂的恐怖袭击是世界各地的威胁。这种攻击的平民幸存者有增加与大脑炎症以及脑细胞死亡有关的问题的风险。平民的治疗将需要开发出可以在沙林暴露后半小时到几个小时内起作用的药物。此处概述的研究项目旨在测试一种新型药物（Q-VD-OPH），该药物可以抑制负责杀死细胞的酶，即caspase，并减少大脑中免疫细胞的炎症。该项目的积极结果将减少脑细胞死亡和在沙林暴露后48小时14天的炎症。进一步的好处将是一系列炎症蛋白和细胞死亡的特定酶的发展，可用于推动新药的发现。