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.

项目概要-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和推进改善医疗 制止癫痫发作和预防急性中毒造成的长期后果的对策 化学威胁剂，特别是有机磷酸胆碱酯酶抑制剂， 二异丙基氟磷酸盐（DFP）、对氧磷和梭曼，或GABAA受体阻断剂，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）是一个中央资源，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 (Core B）。具体而言，核心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-质谱法（LC-MS）或气相色谱-质谱法（GC-MS）， 并在项目中使用标准溶液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 抗癫痫药物，抗炎药和神经保护剂，以协助所有中心项目的剂量选择和 暴露后的给药时间，以及核心B在新的治疗候选物的优化中。核心A将 与项目2和项目3合作，以确定随后使用的癫痫发作和神经病理学的生物标志物 作为治疗效果的生化测试。代谢组学技术，无论是有针对性的和全球性的，将是 就业。靶向代谢组学将集中在氧脂素和神经类固醇，因为这些信号的表达 在癫痫发作后，代谢途径发生了改变，而全球代谢组学将根据需要用于确定更广泛的 癫痫发作和治疗的生物标志物。此外，核心A将继续改进TETS检测方法 因为目前存在的方法对于药代动力学（PK）分析太不敏感。在第一 在项目期间，Core A开发了一种用于TETS的兔多克隆免疫测定法，该方法将针对高 生产量实验室使用和现场检测。核心A最近开始了开发纳米抗体的工作- 基于分析这些小的、热稳定的试剂生产成本低，并且通常提供有价值的测定 用于检测复杂生物基质中的小分子。使用多克隆抗体和 纳米抗体系统将被优化，并以生物传感器的形式包装在现场可部署的平台上， 现场检测当项目有明确的需要时，对其他暴露生物标志物进行免疫测定， 包括其他神经毒性化学物质及其代谢物。具有专门的核心 为所有项目和核心提供分析支持可提高效率，并确保所有项目和核心之间的一致性。 CounterACT中心的组成部分。