Analytical Chemistry

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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.