Allosteric Drug Discovery using Quantum Cascade Laser based Anisotropic THz Microscope (QCL-ATM)

使用基于量子级联激光的各向异性太赫兹显微镜 (QCL-ATM) 进行变构药物发现

• 批准号: 10259392
• 负责人: Alan Lee
• 金额: $ 25.66万
• 依托单位: LONGWAVE PHOTONICS, LLC
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-01 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10259392
• 关键词: Active Sites; Allosteric Site; Binding; Binding Sites; Biochemical; Biological; Buffaloes; Collaborations; Crystallization; Detection; Development; Dihydrofolate Reductase Inhibitor; Elements; Engineering; Ensure; Fingerprint; Frequencies; Fructose; Glucose; HIV-1; Human; Industrialization; Lasers; Life; Measurement; Measures; Mechanics; Methods; Microscope; Microscopy; Modeling; Molecular; Muramidase; Mutation; National Institute of General Medical Sciences; Noise; Optics; Parasites; Pharmaceutical Preparations; Phase; Polishes; Preparation; Protein Dynamics; Proteins; Radiation; Research; Research Personnel; Resistance development; Reverse Transcriptase Inhibitors; Sampling; Signal Transduction; Site; Small Business Technology Transfer Research; Source; Specificity; Spectrum Analysis; Structure; Sucrose; System; Techniques; Temperature; United States National Institutes of Health; Work; absorption; base; detector; drug development; drug discovery; inhibitor/antagonist; innovation; instrument; operation; photonics; prototype; quantum; response; small molecule; thymidylate synthase-dihydrofolate reductase; transmission process; vibration

Allosteric Drug Discovery using Quantum Cascade Laser based Anisotropic THz Microscope (QCL-ATM) Proposal in Response to NIH/NIGMS STTR PA-20-265 This STTR will result in a commercially viable instrument that will enable critical research in allosteric drugs and protein dynamics. To date allosteric inhibitors are largely found serendipitously. Anisotropic THz microspectroscopy (ATM) uniquely measures the long range structural vibrations which serve as a mechanism for allosteric control. ATM provides a tagless means to experimentally determine allosteric target sites. There are NO commercial methods that provide this information currently. ATM systems used to establish the technique are not accessible to a standard biochemical lab. In this STTR we will develop a compact system for turnkey operation by academic and industrial researchers. This will be achieved by a collaboration of optical engineers and biological physicists with unique expertise required. The system requires 1) high power tunable THz source; 2) THz optical system for micro spectroscopy with polarization control; 3) high sensitivity room temperature detection integrated into the microspectroscopy system; and 4) easy user interface. The LongWave Photonics group has innovated high power compact THz sources (quantum cascade lasers, QCL’s) and turn-key measurement systems based on these sources. The Markelz group at UB has innovated ATM. The system, QCL-ATM, will be a turnkey tabletop instrument. In phase I the QCL-ATM will be developed to directly probe vibrations within molecular standards (e.g. sucrose, fructose, and glucose and the protein crystal tetragonal lysozyme) using polarized THz radiation within the 1.6 – 4.3 THz range of our QCL source and measure the change in absorbance as the relative orientation of the crystal molecular samples and polarization axis is varied. Using this demonstration of an integrated QCL-ATM instrument, we will identify the optical and mechanical tolerances associated with the need to place both the sample and the detector entirely within the near-field region of a focused THz beam as preparation for Phase II which will include measurement of the effect of allosteric drugs on protein vibrations and the development of an automated polarization control module and automated multi-sample platen with repeatable high-precision sample alignment to the interrogating THz beam. The specific aims for Phase I are: Aim 1. Construct and Characterize throughput QCL-ATM Microscope in the Far-field. Aim 2. Characterize anisotropic absorbance with the QCL-ATM for molecular crystal standards. Aim 3. Integrate near field pyroelectric detection into the QCL-ATM prototype and establish equivalence to existing systems by measuring spectra of protein crystal

量子级联激光用于变构药物的发现 基于各向异性太赫兹显微镜(QCL-ATM) 针对NIH/NIGMS STTRPA-20-265的建议 这一STTR将产生一种商业上可行的仪器，使变构药物的关键研究成为可能 和蛋白质动力学。迄今为止，变构抑制剂在很大程度上是偶然发现的。各向异性太赫兹 显微光谱学(ATM)独一无二地测量长程结构振动，作为一种机制 用于变构控制。ATM提供了一种无标记的方法来实验确定变构靶点。那里 目前还没有提供这种信息的商业方法。ATM系统用于建立 这项技术是标准的生化实验室无法获得的。在本STTR中，我们将开发一个紧凑的系统，用于 学术和工业研究人员的交钥匙操作。这将通过光学公司的合作来实现 需要具有独特专业知识的工程师和生物物理学家。系统需要1)高功率可调 太赫兹光源；2)偏振控制的太赫兹显微光谱光学系统；3)高灵敏度房间 将温度检测集成到显微光谱系统中；以及4)方便的用户界面。这个 长波光子组发明了高功率紧凑型太赫兹光源(量子级联激光器，QCL) 以及基于这些来源的交钥匙测量系统。UB的Markelz团队发明了自动取款机。这个 QCL-ATM系统将是一个交钥匙的桌面工具。在第一阶段，QCL-ATM将发展为直接 分子标准(如蔗糖、果糖、葡萄糖和蛋白质晶体)内的探针振动 四方溶菌酶)在我们的QCL源的1.6-4.3太赫兹范围内使用偏振太赫兹辐射 测量晶体分子样品的相对取向和偏振时吸光度的变化 轴是不同的。使用此集成QCL-ATM仪器的演示，我们将识别光学和 与需要将样品和检测器完全放置在 聚焦太赫兹光束的近场区域，作为第二阶段的准备，该阶段将包括测量 变构药物对蛋白质振动的影响及自动偏振控制的发展 模块和自动多样品压板，具有可重复的高精度样品对齐 询问太赫兹波束。第一阶段的具体目标是： 目的1.构建和表征QCL-ATM远场通过量显微镜。 目的2.用QCL-ATM表征分子晶体标准品的各向异性吸光度。 目标3.将近场热释电探测集成到QCL-ATM原型中，并建立等价于 现有的通过测量蛋白质晶体光谱的系统