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.

一种专门用于阿尔茨海默病和癌症研究的H/F/X/Y Fast-MAS NMR探针 摘要 超过20%的现有药物（以及更大比例的开发中药物）是含氟的 （包括百忧解、立普妥和环丙沙星等畅销药）。过去十年的稳步进展 显示魔角旋转（MAS）固态NMR（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/19 F/X/2 H探针最近变得更容易获得，并且已经证明这些探针是非常有用的。 对于活性物质的鉴定和表征（使用1H/15 N、19 F/13 C和19 F/2 H/15 N方法）有价值 片段，它们与可溶性蛋白质的结合，以及它们对这种蛋白质-蛋白质相互作用的影响。问题 这种方法对不溶性蛋白质不起作用--例如聚集体和纤维，它们是蛋白质的核心， 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。的 β淀粉样蛋白（Aβ）级联假说开始使神经退行性疾病领域统一起来， 而是理解聚集体进展和初始种子阶段之后的治疗的关键工具 还没有。 适用于含氟药物及其相互作用的多通道研究的MAS探针 由于现有的射频电路在处理上的困难， 在高频率下接近共振。在第一阶段，我们证明了一种新颖的单线圈电路， 允许，第一次，有效的高场H/F/X/Y MAS，在500 MHz的NMR数据和台架实验 和800 MHz的全波详细建模。 第二阶段的提案寻求资金，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19 F检测进行了优化， 在任何或所有其他通道上进行辐照或检测，适用于7-28 T磁场的MAS，带转子 分析表明，在0.7-3 mm的直径范围内， 许多MAS实验是从J-耦合（这不是平均的MAS），以异核和自旋- 相关效应-热梯度、轴向振动和磁性。同时解耦的能力 19 F检测期间，在优化了电路的高分辨率的旋转器中使用快速MAS进行1H、2 H和13 C或15 N检测 和探头设计兼容B 0高达1200 MHz将允许显着增加频谱分辨率， 对例如淀粉样蛋白组装体及其前体聚集体中的19 F标记配体或19 F标记配体的敏感性 标记的DNA-致癌物加合物。 新的探针将允许强大的NMR采集和自动化结构套件 为溶液NMR开发的测定协议，主要依赖于间接检测的三重和四重， 共振方案，成功地应用于刚性氟化样品小于一毫克。探针 将与自动样品交换和样品温度从90 K到420 K兼容。此外，委员会认为， 它将基本上没有所有主要核素（1H，19 F，31 P，13 C，2 H， 15 N和17 O），并且它可以根据几乎所有感兴趣的组合进行调整，从而使其在以下方面也具有无价的价值 如新陈代谢、材料科学、催化和可持续能源等领域。 关键词：氟核磁共振，阿尔茨海默氏症，淀粉样蛋白，MAS，蛋白质聚集体，四共振。