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检测的高通量基础设施 质谱学是一种功能强大、应用广泛的定性和定量分析方法。 对所有类型和大小的化合物进行分析。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。考虑到有能力定位 相对于质谱计的一系列样品，DESI-MS成为高通量(HT)化学物质 分析方法。MS作为一种分析技术的力量是众所周知的，但很少有人意识到 MS也可以作为一种制备方法，例如，它可以用来在表面沉积质量选择的离子 在真空中创造新材料。对有机合成非常重要的是，DESI的一个独特特征是，在 撞击，用于分析反应混合物的溶剂喷雾产生二次微滴，其中 在到达质谱计的途中，反应可能会加速。正是这一显著的特点使Desi- MS是一种强大的合成方法，结合了内置的分析能力。在DARPA的支持下，我们建立了 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 每秒混合。我们现在提议普渡大学和NCATS之间的校内校外合作 ASPIRE实验室。协作的UG3组件将专注于设计、制造和测试 改进的基于DESI-MS的高通量反应筛选系统系统将复制 现有普渡系统的能力，还包括结合小规模合成的新能力 用高通量生物检测。在拟议研究的UH3阶段，该系统将移交给NCATS 并与内部小组合作使用。作为新的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， 目前哪些化疗药物显示出有限的疗效。具体地说，这一努力将需要大规模 潜在胆固醇磺基转移酶(SULT2B1b)的筛选和合成 靶标)抑制性化合物，与生物活性支架的后期功能化一起产生 种类繁多的类比。通过这一努力，该系统将被建立为下一代一体化系统 药物发现平台，具有集成的筛选、合成和生物检测能力。在后者期间 在UH3阶段的阶段，该系统的通用性将在几个其他生物应用中进行测试， 包括乙酰胆碱酯酶再活化剂的定向进化和功能化。成功完成 这些任务将证明新构建的高吞吐量Desi-MS平台是一种有效的方法 用于发现和扩大目前未被下药的生物目标的化学空间。