An H/F/X/Y Fast-MAS NMR Probe Particularly for Alzheimer's and Cancer Research

特别适用于阿尔茨海默病和癌症研究的 H/F/X/Y Fast-MAS NMR 探针

• 批准号: 10224643
• 负责人: Francis DAVID Doty
• 金额: $ 98.33万
• 依托单位: DOTY SCIENTIFIC, INC.
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-04-01 至 2022-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10224643
• 关键词: 3-Dimensional; Adsorption; Alzheimer' s Disease; Amyloid beta-Protein; Area; Binding; Biological; Biological Availability; Biophysics; Caliber; Catalysis; Collaborations; Computer Simulation; DNA Adducts; Data; Deposition; Detection; Development; Diffusion; Disease; Excretory function; Failure; Fluorine; Fluorocarbon Polymers; Fluoxetine; Frequencies; Funding; Industrialization; Institutes; Label; Laboratories; Lead; Ligands; Liquid substance; Magic; Magnetism; Malignant Neoplasms; Measures; Mediating; Medical; Metabolism; Methods; Modeling; Molecular; Molecular Structure; Morphologic artifacts; Neurodegenerative Disorders; Nuclear Magnetic Resonance; Nuclear Proteins; Parkinson Disease; Peptides; Performance; Pharmaceutical Preparations; Pharmacologic Substance; Phase; Prions; Property; Proteins; Protocols documentation; RF coil; Research Personnel; Resolution; Roentgen Rays; Sampling; Scheme; Seeds; Signal Transduction; Silver; Site; Solvents; Speed; Structure; Techniques; Technology; Temperature; Testing; Time; Weight; Work; amyloid formation; analog; analytical tool; anticancer research; atorvastatin; base; design; design and construction; drug candidate; drug development; experimental study; improved; instrument; interest; irradiation; macromolecule; magnetic field; materials science; milligram; novel; peptide drug; protein aggregation; protein data bank; protein protein interaction; prototype; silicon nitride; simulation; solid state nuclear magnetic resonance; success; tool; vibration; virtual

An H/F/X/Y Fast-MAS NMR Probe Particularly for Alzheimer’s and Cancer Research Abstract More than 20% of current drugs (and a much greater fraction of those in development) are fluorinated (including such block-busters as Prozac, Lipitor, and Ciprobay). Steady progress over the past decade has shown magic angle spinning (MAS) solid-state NMR (ssNMR) to be arguably the most powerful analytical tool for studying macro-molecular structures and their dynamics. For solution NMR, four-channel multinuclear 1H/19F/X/2H probes have recently become more readily available, and such have proven to be extremely valuable for identification and characterization (using 1H/15N, 19F/13C, and 19F/2H/15N methods) of active fragments, their binding to soluble proteins, and their effects on such protein-protein interactions. The problem is that such methods don’t work with insoluble proteins – such as the aggregates and fibrils that are central to Alzheimer’s Disease (AD), Parkinson’s Disease (PD), and probably even prion-mediated diseases. The amyloid beta (Aβ) cascade hypothesis is beginning to bring unity to the field of neurodegenerative diseases, but a key tool for understanding aggregate progression and treatment beyond the stages of the initial seeds has not been available. MAS probes suitable for the needed multi-channel studies of fluorinated drugs and their interactions have not been available for high-field NMR instruments because of the difficulties of prior rf circuits in handling close resonances at high frequencies. During the Phase-I, we demonstrated that a novel single-coil circuit permits, for the first time, efficient high-field H/F/X/Y MAS, with NMR data at 500 MHz and bench experiments and full-wave detailed modeling at 800 MHz. This Phase-II proposal seeks funding to continue the development and testing of high-field H/F/X/Y fast-MAS probes based on a novel “single-coil” rf circuit optimized for 19F detection with simultaneous irradiation or detection on any or all of the other channels, and suitable for MAS at fields from 7-28 T, with rotor diameters from 0.7-3 mm. Analysis suggests that a substantial portion of the spectral line broadening seen in many MAS experiments is from J-couplings (which is not averaged by MAS) to heteronuclei and spinner- dependent effects – thermal gradients, axial vibration, and magnetism. The ability to simultaneously decouple 1H, 2H, and 13C or 15N during 19F detection with fast-MAS in a spinner optimized for high resolution with a circuit and probe design compatible with B0 up to 1200 MHz will permit a dramatic increase in spectral resolution and sensitivity on, for example, 19F-labeled ligands in amyloid assemblies and their precursor aggregates, or in 19F labeled DNA-carcinogen adducts. The novel probe would allow the powerful suite of NMR acquisition and automated structure determination protocols developed for solution NMR, which rely mostly on indirect-detected triple- and quad- resonance schemes, to be successfully applied to rigid fluorinated samples smaller than a milligram. The probe will be compatible with automated sample exchange and sample temperatures from 90 K to 420 K. Moreover, it will be essentially devoid of problematic background signals for all the primary nuclides (1H, 19F, 31P, 13C, 2H, 15N, and 17O), and it will be tunable to virtually all combinations of interest, thereby making it also invaluable in such areas as metabolism, materials science, catalysis, and sustainable energy. Key words: fluorine NMR, Alzheimer’s, amyloids, MAS, protein aggregates, quad-resonance.

特别适用于阿尔茨海默病和癌症研究的 H/F/X/Y Fast-MAS NMR 探针 抽象的 超过 20% 的现有药物（以及更大比例的正在开发的药物）是氟化的 （包括百忧解、立普妥和 Ciprobay 等重磅药物）。过去十年的稳步进展 表明魔角旋转 (MAS) 固态核磁共振 (ssNMR) 可以说是最强大的分析工具 用于研究大分子结构及其动力学。对于溶液 NMR，四通道多核 1H/19F/X/2H 探头最近变得更加容易获得，事实证明此类探头非常适合 对于活性物质的识别和表征（使用 1H/15N、19F/13C 和 19F/2H/15N 方法）非常有价值 片段，它们与可溶性蛋白质的结合，以及它们对蛋白质-蛋白质相互作用的影响。问题 是这样的方法不适用于不溶性蛋白质——例如对于蛋白质至关重要的聚集体和原纤维 阿尔茨海默病 (AD)、帕金森病 (PD)，甚至可能是朊病毒介导的疾病。这 β 淀粉样蛋白 (Aβ) 级联假说开始为神经退行性疾病领域带来统一， 而是了解初始种子阶段之外的总体进展和治疗的关键工具 尚未可用。 MAS 探针适用于氟化药物及其相互作用所需的多通道研究 由于先前的射频电路在处理方面存在困难，因此尚未可用于高场核磁共振仪器 高频下的紧密共振。在第一阶段，我们证明了一种新颖的单线圈电路 首次允许高效的高场 H/F/X/Y MAS，具有 500 MHz 的 NMR 数据和台架实验 以及 800 MHz 的全波详细建模。 该第二阶段提案寻求资金以继续开发和测试高场 H/F/X/Y 基于新型“单线圈”射频电路的快速 MAS 探头，针对 19F 检测进行了优化，同时 任何或所有其他通道上的辐照或检测，适用于 7-28 T 场的 MAS，带转子 直径从0.7-3毫米。分析表明，光谱线展宽的很大一部分出现在 许多 MAS 实验是从 J 耦合（未通过 MAS 平均）到异核和自旋- 相关效应——热梯度、轴向振动和磁性。同时解耦的能力 19F 检测期间的 1H、2H 和 13C 或 15N，在旋转器中使用快速 MAS，针对电路的高分辨率进行了优化 与高达 1200 MHz 的 B0 兼容的探头设计将大幅提高光谱分辨率和 例如，淀粉样蛋白组装体及其前体聚集体中或 19F 中 19F 标记的配体的敏感性 标记的DNA-致癌物加合物。 新型探针将提供强大的 NMR 采集和自动化结构套件 为溶液核磁共振开发的测定方案，主要依赖于间接检测的三重和四重 共振方案，成功应用于小于一毫克的刚性氟化样品。探头 将与自动样品交换和 90 K 至 420 K 的样品温度兼容。此外， 对于所有主要核素（1H、19F、31P、13C、2H、 15N 和 17O），并且它几乎可以调整到所有感兴趣的组合，从而使其在以下领域也具有无价的价值： 新陈代谢、材料科学、催化和可持续能源等领域。 关键词：氟核磁共振，阿尔茨海默病，淀粉样蛋白，MAS，蛋白质聚集体，四重共振。