High throughput infrastructure for reaction screening and bioassays

用于反应筛选和生物测定的高通量基础设施

• 批准号: 10447513
• 负责人: Robert Graham Cooks
• 金额: $ 89.83万
• 依托单位: PURDUE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-01 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10447513
• 关键词: Acetylcholinesterase; Address; Air; Bacteria; Biological; Biological Assay; Biological Testing; Cancer Biology; Chemicals; Collaborations; Collection; Computer software; Deposition; Development; Directed Molecular Evolution; Effectiveness; Electrospray Ionization; Enzyme Kinetics; Enzymes; Evaluation; Extramural Activities; In Vitro; Infrastructure; Ions; Joints; Laboratories; Lead; Libraries; Malignant neoplasm of prostate; Mass Spectrum Analysis; Measurement; Measures; Methods; National Center for Advancing Translational Sciences; Organic Synthesis; Outcome; Performance; Phase; Positioning Attribute; Preparation; Property; Qualitative Methods; Reaction; Reagent; Research; Research Personnel; Route; Sampling; Scientist; Solvents; Spectrometry, Mass, Electrospray Ionization; Speed; Sulfate; Surface; System; Techniques; Technology; Technology Transfer; Testing; Training; Use Effectiveness; analog; analytical method; base; bioactive scaffold; chemotherapeutic agent; cholesterol sulfotransferase; cost; data acquisition; design; drug candidate; drug discovery; experimental study; high throughput screening; improved; insight; instrument; instrumentation; interest; ion source; ionization technique; machine learning method; mass spectrometer; medical countermeasure; milligram; nanoscale; nerve agent; new technology; next generation; novel; novel therapeutics; reaction rate; robotic system; scaffold; screening; therapeutic development

High throughput infrastructure for reaction screening and bioassays Mass spectrometry (MS) is a powerful and widely applicable analytical method for qualitative and quantitative analysis of compounds of all types and sizes. Desorption electrospray ionization (DESI) is an ambient ionization method in which samples are analyzed in the open air by impact of primary droplets. Given the ability to position an array of samples relative to the mass spectrometer, DESI-MS becomes a high throughput (HT) chemical analysis method. The power of MS as an analytical technique is well known but it is less commonly realized that MS can also serve as a preparative method, e.g. it can be used to deposit mass-selected ions on surfaces to create new materials in vacuo. Of significant interest to organic synthesis, a unique feature of DESI is that, upon impact, the spray of solvent used to analyze a reaction mixture generates secondary microdroplets in which reactions may be accelerated en route to the mass spectrometer. It is this remarkable feature that makes DESI- MS a powerful synthetic method combined with a built-in analytical capability. With support of DARPA, we built a high throughput system at Purdue capable of automated reaction screening at a rate greater than 1 reaction mixture per second. We now propose an intramural - extramural collaboration between Purdue and the NCATS ASPIRE laboratory. The UG3 component of the collaboration will focus on designing, fabricating, and testing an improved high throughput system for reaction screening based on DESI-MS. The system will replicate the capabilities of the existing Purdue system and also include new capabilities for small-scale synthesis combined with high throughput bioassays. In the UH3 phase of the proposed study, the system will be transferred to NCATS and used in collaboration with the intramural group. As an initial demonstration of the new high throughput platform capabilities, we will pursue the discovery of novel therapeutics for advanced-stage prostate cancer, for which current chemotherapeutic agents show limited effectiveness. Specifically, this effort will entail large-scale screening and synthesis of potential cholesterol sulfotransferase (SULT2B1b, a currently undrugged biological target) inhibitory compounds, together with late-stage functionalization of bioactive scaffolds to generate a diverse range of analogs. Through this effort, the system will be established as an all-in-one next-generation drug discovery platform, with integrated screening, synthesis, and biological assay capabilities. During the latter stages of the UH3 phase, the versatility of the system will be tested in several other biological applications, including directed evolution and functionalization of acetylcholinesterase reactivators. Successful completion of these tasks will demonstrate the newly constructed high throughput DESI-MS platform to be an efficient method for the discovery and expansion of chemical space towards currently undrugged biological targets.

用于反应筛选和生物测定的高通量基础设施 质谱 (MS) 是一种强大且广泛适用的定性和定量分析方法 分析所有类型和大小的化合物。解吸电喷雾电离 (DESI) 是一种环境电离 在露天通过初级液滴的撞击对样品进行分析的方法。考虑到定位能力 相对于质谱仪的一系列样品，DESI-MS 成为高通量 (HT) 化学品 分析方法。 MS 作为一种分析技术的强大功能众所周知，但很少有人意识到 MS 也可以作为一种制备方法，例如它可用于在表面上沉积质量选择的离子 在真空中创造新材料。 DESI 对有机合成具有重要意义，其独特之处在于， 影响，用于分析反应混合物的溶剂喷雾会产生次级微滴，其中 反应在前往质谱仪的途中可能会加速。正是这个显着的特点使得 DESI- MS 是一种与内置分析功能相结合的强大合成方法。在 DARPA 的支持下，我们建造了 普渡大学的高通量系统能够以大于 1 个反应的速率进行自动反应筛选 每秒的混合物。我们现在提议普渡大学和 NCATS 之间进行校内校外合作 ASPIRE实验室。合作的 UG3 部分将重点关注设计、制造和测试 改进的基于 DESI-MS 的反应筛选高通量系统。系统将复制 现有普渡系统的功能，还包括小规模合成的新功能 具有高通量生物测定。在拟议研究的UH3阶段，系统将转移到NCATS 并与校内小组合作使用。作为新高吞吐量的初步演示 平台能力，我们将致力于发现晚期前列腺癌的新疗法， 目前的化疗药物显示出有限的效果。具体来说，这项工作将需要大规模 筛选和合成潜在的胆固醇磺基转移酶（SULT2B1b，一种目前未成药的生物酶） 目标）抑制性化合物，以及生物活性支架的后期功能化，以产生 各种类似物。通过这一努力，该系统将成为一个一体化的下一代 药物发现平台，具有集成的筛选、合成和生物测定功能。后者期间 UH3 阶段，系统的多功能性将在其他几个生物应用中进行测试， 包括乙酰胆碱酯酶再激活剂的定向进化和功能化。顺利完成 这些任务将证明新构建的高通量 DESI-MS 平台是一种有效的方法 发现和扩展化学空间以实现目前未药物化的生物目标。