Analytical Chemistry

分析化学

• 批准号: 10907371
• 负责人: Christopher Arthur LeClair
• 金额: $ 250.06万
• 依托单位: NATIONAL CENTER FOR ADVANCING TRANSLATIONAL SCIENCES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10907371
• 关键词: Acceleration; Acoustics; Address; Adopted; Adoption; Aliquot; Analytical Chemistry; Antiviral Agents; Area; Bar Codes; Binding Proteins; Biological; Biology; Biotin; COVID-19 pandemic; COVID-19 treatment; Calibration; Chemicals; Chemistry; Cholesterol; Chromatography; Collaborations; Collection; Communities; Computer software; Coronavirus; Custom; Data; Data Analyses; Data Storage and Retrieval; Detection; Development; Disease; Emerging Technologies; Ensure; Enzymes; Equipment; Equipment and supply inventories; Extramural Activities; Family; Goals; Heart Diseases; High Pressure Liquid Chromatography; Hybrids; Industry Standard; Informatics; Intellectual Property; Kidney Diseases; Laboratories; Libraries; Lipids; Liquid Chromatography; Liquid substance; Maintenance; Management Information Systems; Manuals; Mass Fragmentography; Mass Spectrum Analysis; Medicine; Mentorship; Methods; Mission; Modality; Monitor; NMR Spectroscopy; National Center for Advancing Translational Sciences; Nuclear Magnetic Resonance; Oral; Peptides; Performance; Pharmaceutical Chemistry; Phosphatidylcholine-Sterol O-Acyltransferase; Phosphotransferases; Preparation; Process; Productivity; Proteins; Protocols documentation; Publications; Reaction; Reproducibility; Research; Research Personnel; Research Project Grants; Resolution; Resources; Sampling; Schedule; Scientist; Services; Site; Supercritical Fluid Chromatography; System; Techniques; Technology; Therapeutic; Time; Translational Research; Tube; Vendor; Vibrational Circular Dichroism; Viral; Virus; Water; Work; advanced analytics; analytical method; antiviral drug development; assay development; coronavirus disease; data acquisition; data management; density; design; drug development; drug discovery; experimental analysis; high throughput screening; improved; innovation; instrument; instrumentation; interest; lipidomics; liquid chromatography mass spectrometry; meetings; member; metabolomics; milligram; multiple omics; novel; pandemic disease; pandemic potential; pre-clinical; preclinical development; prevent; process optimization; productivity loss; professional atmosphere; programs; screening; screening program; small molecule; small molecule inhibitor; success; telework; tool; translational scientist

The ACC continued to navigate the logistical challenge of providing core analytical chemistry services to DPI researchers within the framework of the current hybrid work environment. To accommodate the varied onsite and telework schedules of DPI staff, ACC team members worked together to ensure the continuous operability of all service lines and fulfillment of all responsibilities to maintain productivity. As such, the ACC successfully maintained resources and provided analytical support for scientists across the 3 DPI scientific branches (ETB, TDB, and CGB). Even with the end of the COVID pandemic, the ACC has maintained involvement in COVID-related projects primarily through support of the Antiviral Program for Pandemics (APP) team whose mission is to accelerate antiviral development through early discovery and preclinical development of safe and effective oral antivirals. The initial priority for the APP is to develop treatments for SARS-CoV-2 and other coronaviruses, with the program expanding to address other virus families with pandemic potential. APP chemists within DPI are tasked with the discovery and development potential small molecule antiviral treatments for a range of viruses that have pandemic potential. Utilizing the Sample Management and Resource Tracking (SMART) Centralized Sample Purification and Processing Platform, the ACC purified, isolated, identified, and analyzed a wide array of chemical modalities as part of drug discovery efforts. The purification employs semi-preparative liquid chromatography systems utilizing UV- and mass-directed detection and collection to purify a wide range of small molecules and peptides in the milligrams to grams scale. The SMART laboratory information management system (LIMS) continues to be utilized for sample submission, compound purification and processing, sample registration, compound inventory, sample tracking, and data retrieval and management. This process for the rapid progression of compounds from Medicinal Chemistry to Compound Management (CoMa) has an average 5-day cycle time and resulted in >2,500 new compounds synthesized by DPI chemistry being added to the NCATS compound library. With the acquisition of new instrumentation to maintain, upgrade, and expand analytical chemistry technology and capabilities, as well as meet the increased demand for purification and analysis, we renovated and reorganized several lab areas including the ACC Chiral Separation Lab and the ACC General Instrument Lab. This renovation included the acquisition of custom laboratory benches, installation of high-density storage shelves, and the surplus of obsolete equipment to maximize usage of the space and improve operational efficiency. We completed the installation and calibration of the newly acquired instrumentation, which included an Agilent G6546 LC-QTOF system, a Waters UPLC-MS system, a Waters AutoPurification system, and an SPT Labtech Lab2Lab pneumatic tubing system. The site preparation for installation of a Waters UPC2-QDa supercritical fluid chromatography (SFC) instrument that will provide additional separation capabilities has also been finished. The ACC is redesigning the NMR Spectroscopy Lab to expand the space for consolidation of instrumentation. This new NMR Lab will include space for six NMR instruments, several automated sample handlers, an automated liquid handling platform, a wet chemistry lab, a meeting room, and an office area for up to 8 staff and trainees. An old NMR instrument has already been decommissioned with subsequent installation of a more advanced Bruker Avance Neo 400 MHz system with minimal loss of productivity. The renovation is estimated to be completed by the end of fiscal year 2024. In anticipation of the new lab, the development of our NMR-based fragment screening program has been steadily progressing. The ACC and CoMa have been creating workflows for the storage and usage of the stock solutions of the chemically diverse fragment library. This includes processes for the transfer and dilution of sample aliquots for screening. The experimental protocols are being validated by performing ligand-protein binding studies to identify reversible small molecule binders of lecithin-cholesterol acyltransferase (LCAT), an enzyme involved in the clearance of excess cholesterol. The goal is to develop activators of LCAT that will improve cholesterol elimination as a means of preventing heart and kidney disease. A key aspect of ACC research is the utilization of LCMS analysis throughout all stages of the workflow for reaction monitoring, identity verification, and purity analysis of the desired compounds. The ACC and ASPIRE have completed the installation of the SPT Labtech Lab2Lab pneumatic sample transport system, which will improve the efficiency and increase productivity of LCMS analysis by rapidly and efficiently sending 2D-barcoded minitubes from five labs across two buildings to a centralized analytical instrumentation hub. This enables ease of oversight and maintenance, sample tracking capabilities and maximization of instrument usage, and freeing of limited laboratory space. This initiative has required the collaboration of two LCMS vendors to enable integration of their instrumentation with the Lab2Lab system. We are also working with the DPI medicinal chemists to change their lab practices and start utilizing the Lab2Lab system. In our continued efforts to advance the ACC sample purification and processing workflow, we finalized the design of our stand-alone automated purification platform and construction of the system has begun. This initiative involves the integration of varied components and software applications, which requires the coordination of various vendors and internal partners. Once completed the automated purification platform will eliminate numerous manual steps in workflow accelerating the drug discovery process and enabling chromatography staff to concurrently work on other research tasks. The design and development of the fully automated purification module that will be integrated within the ASPIRE automated chemistry platform has commenced. Additionally, the ACC and ASPIRE are collaborating in the development of an automated quantitative NMR (qNMR) platform from sample preparation to data acquisition to data analysis. This innovative platform will enable fast and efficient determination of sample concentration, purity analysis, mixture identification, and reaction screening for optimization of conditions. Mass Spectrometry research efforts have been proceeding with great success. The ACC has screened >150,000 compounds as part of numerous Agilent RapidFire MS-based high-throughput assays providing excellent data that has identified small molecule inhibitors and activators for a number of disease-related protein targets. The ACC has continued to work with the DPI Informatics Core (IFX) on a multi-omics pipeline focused on leveraging DPI resources for end-to-end research projects (assay development, experimentation, and data analysis). Advances have been made to adapt our automated high-throughput sample preparation platform to accommodate samples for metabolomics and lipidomics. We are exploring the use of Covaris adaptive focused acoustics (AFA) technology in the homogenization of biological samples, as well as reduction in material amounts used. Finally, the ACC and ChemTech are collaborating to develop an MS-based kinase profiling platform utilizing biotin-tagged ADP and ATP probes.

ACC 继续应对在当前混合工作环境框架内为 DPI 研究人员提供核心分析化学服务的后勤挑战。为了适应 DPI 员工不同的现场和远程工作安排，ACC 团队成员共同努力，确保所有服务线的持续运行并履行所有职责以保持生产力。因此，ACC 成功维护了资源，并为 DPI 3 个科学分支（ETB、TDB 和 CGB）的科学家提供了分析支持。即使新冠病毒大流行已经结束，ACC 主要通过大流行病抗病毒计划 (APP) 团队的支持，继续参与与新冠病毒相关的项目，该团队的使命是通过安全有效的口服药物的早期发现和临床前开发来加速抗病毒药物的开发。抗病毒药物。该 APP 的首要任务是开发针对 SARS-CoV-2 和其他冠状病毒的治疗方法，随后该计划将扩展到解决其他具有大流行潜力的病毒家族。 DPI 内的 APP 化学家的任务是发现和开发针对一系列具有大流行潜力的病毒的潜在小分子抗病毒治疗方法。 ACC 利用样品管理和资源跟踪 (SMART) 集中样品纯化和处理平台，纯化、分离、鉴定和分析了多种化学模式，作为药物发现工作的一部分。纯化采用半制备型液相色谱系统，利用紫外和质量定向检测和收集来纯化毫克至克级的各种小分子和肽。 SMART 实验室信息管理系统 (LIMS) 继续用于样品提交、化合物纯化和处理、样品注册、化合物库存、样品跟踪以及数据检索和管理。该化合物从药物化学到化合物管理 (CoMa) 的快速进展过程平均需要 5 天的周期时间，导致 DPI 化学合成的超过 2,500 种新化合物被添加到 NCATS 化合物库中。 随着购买新仪器来维护、升级和扩展分析化学技术和能力，并满足日益增长的纯化和分析需求，我们对多个实验室区域进行了翻新和重组，包括 ACC 手性分离实验室和 ACC 通用仪器实验室。此次改造包括购买定制实验室工作台、安装高密度储物架以及剩余的陈旧设备，以最大限度地利用空间并提高运营效率。我们完成了新购置仪器的安装和校准，其中包括 Agilent G6546 LC-QTOF 系统、Waters UPLC-MS 系统、Waters AutoPurification 系统和 SPT Labtech Lab2Lab 气动管路系统。安装沃特世 UPC2-QDa 超临界流体色谱 (SFC) 仪器的现场准备工作也已完成，该仪器将提供额外的分离能力。 ACC 正在重新设计核磁共振波谱实验室，以扩大仪器整合的空间。这个新的 NMR 实验室将包括可容纳六台 NMR 仪器的空间、几个自动样品处理机、一个自动液体处理平台、一个湿化学实验室、一间会议室以及一个最多可容纳 8 名员工和学员的办公区。旧的 NMR 仪器已退役，随后安装了更先进的 Bruker Avance Neo 400 MHz 系统，生产力损失最小。改造预计将于 2024 财年年底完成。在新实验室建成的同时，我们基于 NMR 的片段筛选项目的开发一直在稳步推进。 ACC 和 CoMa 一直在创建用于存储和使用化学多样化片段库储备溶液的工作流程。这包括用于筛选的样品等分试样的转移和稀释过程。通过进行配体-蛋白质结合研究来验证实验方案，以确定卵磷脂胆固醇酰基转移酶（LCAT）的可逆小分子结合物，LCAT是一种参与清除过量胆固醇的酶。目标是开发 LCAT 激活剂，改善胆固醇消除，从而预防心脏和肾脏疾病。 ACC 研究的一个关键方面是在工作流程的所有阶段利用 LCMS 分析来进行反应监测、身份验证和所需化合物的纯度分析。 ACC 和 ASPIRE 已完成 SPT Labtech Lab2Lab 气动样品传输系统的安装，该系统将通过快速高效地将 2D 条形码微型管从两栋大楼的五个实验室发送到集中分析仪器中心，提高 LCMS 分析的效率和生产率。这使得监督和维护变得容易，样品跟踪能力和仪器使用最大化，并释放有限的实验室空间。该计划需要两家 LCMS 供应商的合作，以实现其仪器与 Lab2Lab 系统的集成。我们还与 DPI 药物化学家合作，改变他们的实验室实践并开始使用 Lab2Lab 系统。 在我们不断努力推进 ACC 样品纯化和处理工作流程的过程中，我们完成了独立自动化纯化平台的设计，并且系统的建设已经开始。该举措涉及各种组件和软件应用程序的集成，需要各个供应商和内部合作伙伴的协调。一旦完成，自动化纯化平台将消除工作流程中的大量手动步骤，加速药物发现过程，并使色谱工作人员能够同时从事其他研究任务。将集成到 ASPIRE 自动化化学平台中的全自动纯化模块的设计和开发已经开始。此外，ACC 和 ASPIRE 正在合作开发自动化定量 NMR (qNMR) 平台，从样品制备到数据采集再到数据分析。这一创新平台将能够快速有效地测定样品浓度、纯度分析、混合物鉴定和反应筛选以优化条件。 质谱研究工作一直在取得巨大成功。 ACC 已经筛选了超过 150,000 种化合物，作为众多基于安捷伦 RapidFire MS 的高通量测定的一部分，提供了出色的数据，识别了许多疾病相关蛋白质靶标的小分子抑制剂和激活剂。 ACC 继续与 DPI 信息学核心 (IFX) 合​​作开发多组学管道，重点是利用 DPI 资源进行端到端研究项目（测定开发、实验和数据分析）。我们的自动化高通量样品制备平台已取得进展，可以适应代谢组学和脂质组学的样品。我们正在探索使用 Covaris 自适应聚焦声学 (AFA) 技术对生物样品进行均质化，并减少材料用量。最后，ACC 和 ChemTech 正在合作开发一个基于 MS 的激酶分析平台，利用生物素标记的 ADP 和 ATP 探针。

项目成果

Discovery and Optimization of 2H-1λ2-Pyridin-2-one Inhibitors of Mutant Isocitrate Dehydrogenase 1 for the Treatment of Cancer.

突变异柠檬酸脱氢酶 1 的 2H-1α2-Pyridin-2-one 抑制剂的发现和优化用于治疗癌症。

• 发表时间: 2021
• 影响因子: 7.3
• 作者: Rohde, Jason M;Karavadhi, Surendra;Pragani, Rajan;Liu, Li;Fang, Yuhong;Zhang, Weihe;McIver, Andrew;Zheng, Hongchao;Liu, Qingyang;Davis, Mindy I;Urban, Daniel J;Lee, Tobie D;Cheff, Dorian M;Hollingshead, Melinda;Henderson, Mark J;Martinez, N
• 通讯作者: Martinez, N

High-throughput protein modification quantitation analysis using intact protein MRM and its application on hENGase inhibitor screening.

使用完整蛋白 MRM 进行高通量蛋白修饰定量分析及其在 hENGase 抑制剂筛选中的应用。

• 发表时间: 2021-08-15
• 影响因子: 6.1
• 作者: Tao, Dingyin;Xu, Miao;Farkhondeh, Atena;Burns, Andrew P;Rodems, Steven;Might, Matthew;Zheng, Wei;LeClair, Christopher A
• 通讯作者: LeClair, Christopher A

Facile High-Performance Liquid Chromatography Mass Spectrometry Method for Analysis of Cyclocreatine and Phosphocyclocreatine in Complex Mixtures of Amino Acids.

用于分析氨基酸复杂混合物中环肌酸和磷酸环肌酸的简便高效液相色谱质谱法。

• 发表时间: 2019-06-26
• 影响因子: 6.1
• 作者: Tao, Dingyin;Leister, William;Huang, Wenwei;Alimardanov, Asaf;LeClair, Christopher A
• 通讯作者: LeClair, Christopher A

A versatile polypharmacology platform promotes cytoprotection and viability of human pluripotent and differentiated cells.

多功能多药理学平台可促进人类多能和分化细胞的细胞保护和活力。

• 发表时间: 2021
• 影响因子: 48
• 作者: Chen, Yu;Tristan, Carlos A;Chen, Lu;Jovanovic, Vukasin M;Malley, Claire;Chu, Pei;Ryu, Seungmi;Deng, Tao;Ormanoglu, Pinar;Tao, Dingyin;Fang, Yuhong;Slamecka, Jaroslav;Hong, Hyenjong;LeClair, Christopher A;Michael, Sam;Austin, Christoph
• 通讯作者: Austin, Christoph

Discovery of Novel Small-Molecule Scaffolds for the Inhibition and Activation of WIP1 Phosphatase from a RapidFire Mass Spectrometry High-Throughput Screen.

通过 RapidFire 质谱高通量筛选发现新型小分子支架，用于抑制和激活 WIP1 磷酸酶。

• DOI: 10.1021/acsptsci.2c00147
• 发表时间: 2022-09-28
• 期刊: ACS pharmacology & translational science
• 作者: V. Clausse;Yuhong Fang;Dingyin Tao;Harich;ra D. Tagad;ra;Hongmao Sun;Yuhong Wang;Surendra Karavadhi;Kelly C. Lane;Zhen‐Dan Shi;O. Vasalatiy;Christopher A. LeClair;R. Eells;Min Shen;S. Patnaik;E. Appella;N. Coussens;M. Hall;D. Appella
• 通讯作者: D. Appella