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

基本信息

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
项目类别：
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

项目摘要

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

使用量子级联激光进行变构药物发现基于各向异性太赫兹显微镜 (QCL-ATM) 响应 NIH/NIGMS STTR PA-20-265 的提案该 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迄今为止，变构抑制剂主要是偶然发现的。显微光谱学 (ATM) 独特地测量作为一种机制的长程结构振动 ATM 提供了一种通过实验确定变构靶位点的无标签方法。目前没有任何商业方法可以提供用于建立 ATM 系统的信息。标准生化实验室无法使用该技术。在本 STTR 中，我们将开发一个紧凑的系统。学术和工业研究人员的统包操作将通过光学合作来实现。该系统需要具有独特专业知识的工程师和生物物理学家 1) 高功率可调。太赫兹光源；2）具有偏振控制的太赫兹光学系统；3）高灵敏度室；温度检测集成到显微光谱系统中；4) 简单的用户界面。长波光子学小组创新了高功率紧凑型太赫兹源（量子级联激光器，QCL） UB 的 Markelz 团队对 ATM 进行了创新。 QCL-ATM 系统将是一款交钥匙桌面仪器，在第一阶段，QCL-ATM 将直接开发。分子标准（例如蔗糖、果糖和葡萄糖以及蛋白质晶体）内的探针振动四方溶菌酶）使用我们的 QCL 源的 1.6 – 4.3 THz 范围内的偏振太赫兹辐射和测量吸光度随晶体分子样品的相对取向和偏振的变化通过集成 QCL-ATM 仪器的演示，我们将识别光学和轴。与将样品和探测器完全放置在容器内的需要相关的机械公差聚焦太赫兹光束的近场区域，作为第二阶段的准备，其中包括测量变构药物对蛋白质振动的影响和自动偏振控制的发展模块和自动多样品压板，可重复高精度样品对准第一阶段的具体目标是：目标 1. 构建并表征远场吞吐量 QCL-ATM 显微镜。目标 2. 使用 QCL-ATM 表征分子晶体标准品的各向异性吸光度。目标 3. 将近场热释电检测集成到 QCL-ATM 原型中并建立与通过测量蛋白质晶体光谱的现有系统