Analytical Chemistry

分析化学

• 批准号: 10204120
• 负责人: BRUCE D HAMMOCK
• 金额: $ 45.58万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-01 至 2023-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10204120
• 关键词: ADME Study; Acute; Analytical Chemistry; Animals; Anti-Inflammatory Agents; Antibodies; Anticonvulsants; Biochemical; Biological; Biological Assay; Biological Markers; Biosensor; Brain; Chemicals; Cholinesterase Inhibitors; Complex; Detection; Dose; Drug Kinetics; Ensure; Formulation; Goals; Immunoassay; Intoxication; Isoflurophate; Laboratories; Mass Fragmentography; Methods; Mus; Neuroprotective Agents; Organophosphates; Oryctolagus cuniculus; Paraoxon; Pharmaceutical Preparations; Pharmacologic Substance; Picrotoxin; Quality Control; Rattus; Reagent; Recombinants; Resources; Seizures; Signal Pathway; Site; Soman; System; Techniques; Time; Training; Treatment Efficacy; Work; advanced analytics; base; biomarker identification; chemical threat; design; detection assay; detection method; improved; liquid chromatography mass spectrometry; medical countermeasure; metabolomics; nanobodies; neuropathology; neurosteroids; neurotoxic; novel; novel therapeutics; prevent; reagent standard; receptor; small molecule; tetramethylenedisulfotetramine; therapeutic candidate; therapeutic evaluation

Project Summary – Analytical Chemistry Core – Core A The overall goal of the UC Davis CounterACT Center of Excellence is to identify and advance improved medical countermeasures for stopping seizures and preventing long-term consequences resulting from acute intoxication with chemical threat agents, specifically organophosphate cholinesterase inhibitors like diisopropylfluorophosphate (DFP), paraoxon and soman, or GABAA receptor blockers like tetramethylene- disulfotetramine (TETS) or picrotoxin. The Analytical Chemistry Core (Core A) is a central resource designed to provide state-of-the-art, comprehensive analytical support to Projects 1, 2, and 3, and the Probe and Pharmaceutical Optimization Core (Core B). Specifically, Core A will develop methods for the detection of target compounds and their metabolites by liquid chromatography–mass spectrometry (LC-MS) or gas chromatography–mass spectrometry (GC-MS), and provide quality control (QC) analysis of standard solutions prior to their use in projects. Core A will use these methods to perform mouse and rat ADME (absorption, distribution, metabolism, and excretion) studies for antiseizure drugs, anti-inflammatories and neuroprotectants to assist all Center projects in dose selection and time of administration post-exposure, and Core B in the optimization of novel therapeutic candidates. Core A will work with Project 2 and Project 3 to identify biomarkers of seizures and neuropathology to subsequently be used as a biochemical test of therapeutic efficacy. Metabolomics techniques, both targeted and global, will be employed. Targeted metabolomics will focus on oxylipins and neurosteroids, since expression of these signaling pathways are altered after a seizure, while global metabolomics will be employed as needed to identify broader biomarkers of seizure and therapy. Additionally, Core A will continue improving methods for TETS detection since the currently existing methods are too insensitive for pharmacokinetics (PK) analysis. During the first project period, Core A developed a rabbit polyclonal immunoassay for TETS, which will be optimized for high throughput laboratory use and for field detection. Core A has more recently initiated work to develop a nanobody- based assay. These small, heat stable reagents are inexpensive to produce and often provide valuable assays for the detection of small molecules in complex biological matrices. Immunoassays using both the polyclonal and the nanobody system will be optimized, and packaged in biosensor formats in a field deployable platform for on- site detection. When there is a clear need from the projects, immunoassays to other biomarkers of exposure and effect, including other neurotoxic chemicals, and their metabolites, will be created. Having a specialized core providing analytical support for all projects and cores improves efficiency and ensures consistency across all components of the CounterACT Center.

项目摘要 – 分析化学核心 – 核心 A 加州大学戴维斯分校 CounterACT 卓越中心的总体目标是确定并推进改进的医疗 停止癫痫发作和预防急性中毒造成的长期后果的对策 使用化学威胁剂，特别是有机磷酸酯胆碱酯酶抑制剂，例如 二异丙基氟磷酸盐 (DFP)、对氧磷和梭曼，或 GABAA 受体阻滞剂，如四亚甲基- 二磺四胺 (TETS) 或印防己毒素。 分析化学核心（核心 A）是一个中心资源，旨在提供最先进的、 为项目 1、2 和 3 以及探针和药物优化核心提供全面的分析支持 （核心B）。具体来说，Core A 将开发检测目标化合物及其代谢物的方法 通过液相色谱-质谱（LC-MS）或气相色谱-质谱（GC-MS）， 并在标准解决方案用于项目之前对其进行质量控制 (QC) 分析。核心A将使用这些 进行小鼠和大鼠 ADME（吸收、分布、代谢和排泄）研究的方法 抗癫痫药、抗炎药和神经保护剂，协助所有中心项目进行剂量选择和 暴露后的给药时间，以及核心 B 在优化新型候选治疗药物中的作用。核心A将 与项目 2 和项目 3 合作，确定随后使用的癫痫和神经病理学生物标志物 作为治疗效果的生化测试。有针对性的和全球性的代谢组学技术将 受雇。靶向代谢组学将重点关注氧脂质和神经类固醇，因为这些信号的表达 癫痫发作后通路会发生改变，同时将根据需要采用全局代谢组学来识别更广泛的 癫痫发作和治疗的生物标志物。此外，Core A 将继续改进 TETS 检测方法 因为目前现有的方法对于药代动力学（PK）分析太不敏感。第一次期间 项目期间，Core A 开发了一种针对 TETS 的兔多克隆免疫测定法，该测定法将针对高 高通量实验室使用和现场检测。 Core A 最近启动了纳米抗体的开发工作 为基础的测定。这些小型、热稳定的试剂生产成本低廉，并且通常提供有价值的检测 用于检测复杂生物基质中的小分子。使用多克隆和 纳米抗体系统将被优化，并以生物传感器格式封装在可现场部署的平台中，以用于现场部署 现场检测。当项目有明确需要时，对其他暴露和生物标志物进行免疫测定 将会产生影响，包括其他神经毒性化学物质及其代谢物。拥有专业的核心 为所有项目和核心提供分析支持，提高效率并确保所有项目的一致性 CounterACT 中心的组成部分。