Development of a transformative approach to the rapid detection, isolation, and i

开发一种快速检测、隔离和检测的变革性方法

• 批准号: 8418699
• 负责人: Birgit Ursula Jaki
• 金额: $ 19.49万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-02-15 至 2015-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8418699
• 关键词: AIDS/HIV problem; Actinobacteria class; Affect; Agar; Antibiotics; Antitubercular Agents; Back; Bacterial Infections; Biological; Biological Factors; Categories; Cephalosporins; Chemical Structure; Chromatography; Collaborations; Collection; Communities; Data; Databases; Detection; Developing Countries; Development; Diffusion; Disease Management; Drug resistance; Drug resistance in tuberculosis; Erythromycin; Evaluation; Extreme drug resistant tuberculosis; Fermentation; Fingerprint; Fractionation; Glean; Goals; Growth; Investigation; Knowledge; Lead; Libraries; Magnetism; Maps; Methodology; Methods; Microbiology; Minor; Molecular Weight; Multi-Drug Resistance; Mycobacterium tuberculosis; Natural Product Drug; Nature; Nuclear; Organism; Performance; Pharmaceutical Preparations; Population; Procedures; Protocols documentation; Public Health; Regimen; Research; Rifampin; Sampling; Scheme; Signal Transduction; Soil; Solutions; South Korea; Spectrum Analysis; Spottings; Staging; Standardization; Streptomycin; Structure; Technology; Testing; Tetracyclines; Thin Layer Chromatography; Time; Tuberculosis; United States National Institutes of Health; Universities; antimicrobial; base; chemical property; drug development; drug discovery; fungus; global health; innovative technologies; interest; luminescence; microorganism; new technology; novel; novel strategies; pathogen; rapid detection; resistant strain; screening; success; treatment duration; tuberculosis drugs; two-dimensional; vector

DESCRIPTION (provided by applicant): The challenging disease management of tuberculosis (TB), a bacterial disease caused by Mycobacterium tuberculosis, and the increasing abundance of multi/extensive drug-resistant strains has precipitated to a severe public health problem. While global this mostly affects developing countries and regions with prevalent HIV/AIDS problems, and leads to an urgent need to find novel anti-TB lead compounds. The lack of success in finding such compounds by screening synthetic libraries directs the focus of interest back to the chemically diverse natural products. The constraints of classical bioactivity guided isolation are labor intensity, time commitment, and most important, the difficulty in detecting, isolating, and characterizing bioactive minor constituents. The present exploratory project will lay the groundwork for a transformative concept to the rapid detection, isolation, and identification of anti-tuberculosis natural products. The technology will be developed on extracts of cultured actinomycete strains and will be a prerequisite for a large-scale anti- TB drug discovery project on a panel of pre-selected anti-TB active actinomycete strains. A novel TLC-MS-(TB)-bioautography method will be developed for the straight-forward identification of active key compounds in an analytical setting that allows the mass spectrometric analysis of planar chromatographic sample separations and in parallel the evaluation of their biological activity profile. Combining the mass spectroscopic with biological activity data will generate two-dimensional maps that directly correlate the anti-TB activity with structural information. The isolation procedure can significantly be shortened by using the knowledge of the chemical properties of the active principle in an early stage of the separation procedure. A specific protocol can be tailored with counter current chromatography based on the polarity based GUESS principle and/or to directly upscale to preparative TLC. State-of-the-art technology in planar chromatography (high-performance TLC, automated "spray-on" sampling, gradient elution, semi-automatic extraction), microbiology (TB agar-overlay with luminescence detection), and MS spectroscopy (LC-MS2) will be used to target the isolation of lead compounds. Known chemical structures will be dereplicated based on their molecular weight, as gleaned from MS (and LC-MS2) and EI database (NIST, Wiley) search. Full structure elucidation of new or otherwise important isolated active principles will utilize state-of-the-art 1D/2D NMR- and MS-based structural information and will be supported by novel methodology like cryoprobe NMR, nuclear fingerprinting and purity-activity relationships based on quantitative NMR and modern anti-TB activity assessment. TLC-MS-(TB)-bioautography will be compared with established technologies, optimized, and validated. The concept can generally be applied in natural product drug discovery to handle large quantities of samples and will in our case be applied to pre-selected anti-TB active actinomycete strains. We expect that application of this novel technology will result in the discovery of a plethora of new, heretofore unidentified anti-TB natural products and that one or more of these will be a viable lead compound for TB drug development.

描述（由申请人提供）：结核病（TB）是一种由结核分枝杆菌引起的细菌性疾病，其疾病管理具有挑战性，并且多重/广泛耐药菌株的数量不断增加，已导致严重的公共卫生问题。虽然在全球范围内，这主要影响艾滋病毒/艾滋病问题普遍存在的发展中国家和地区，并导致迫切需要寻找新型抗结核先导化合物。通过筛选合成文库未能成功找到此类化合物，这使得人们的兴趣焦点又回到了化学多样性的天然产物上。经典生物活性引导分离的限制是劳动强度、时间投入，最重要的是检测、分离和表征生物活性次要成分的困难。目前的探索性项目将为快速检测、分离和鉴定抗结核天然产物的变革性概念奠定基础。该技术将在培养的放线菌菌株的提取物上进行开发，并将成为针对一组预先选择的抗结核活性放线菌菌株进行大规模抗结核药物发现项目的先决条件。将开发一种新型 TLC-MS-(TB)-生物自记录方法，用于在分析环境中直接鉴定活性关键化合物，该分析环境允许对平面色谱样品分离进行质谱分析，并并行评估其生物活性概况。将质谱与生物活性数据相结合将生成二维图谱，将抗结核活性与结构信息直接关联起来。通过在分离过程的早期阶段利用活性成分的化学性质的知识，可以显着缩短分离过程。可以使用基于极性的 GUESS 原理的逆流色谱法定制特定的方案和/或直接升级为制备型 TLC。平面色谱（高性能薄层色谱、自动“喷雾”采样、梯度洗脱、半自动提取）、微生物学（带发光检测的 TB 琼脂覆盖层）和 MS 光谱（LC -MS2) 将用于目标分离先导化合物。已知的化学结构将根据其分子量进行重复，如从 MS（和 LC-MS2）和 EI 数据库（NIST、Wiley）搜索中收集到的。新的或其他重要的分离活性成分的完整结构阐明将利用最先进的基于 1D/2D NMR 和 MS 的结构信息，并将得到冷冻探针 NMR、核指纹和基于纯度-活性关系等新颖方法的支持。定量核磁共振和现代抗结核活性评估。 TLC-MS-(TB)-生物自显影将与现有技术进行比较、优化和验证。该概念通常可应用于天然产物药物发现以处理大量样品，并且在我们的案例中将应用于预先选择的抗结核活性放线菌菌株。我们预计，这项新技术的应用将导致发现大量迄今为止尚未鉴定的新型抗结核天然产物，并且其中一种或多种将成为结核病药物开发的可行先导化合物。

项目成果

Bioautography with TLC-MS/NMR for Rapid Discovery of Anti-tuberculosis Lead Compounds from Natural Sources.

使用 TLC-MS/NMR 进行生物自显影，可快速发现天然来源的抗结核先导化合物。

• 发表时间: 2016-04-08
• 影响因子: 5.3
• 作者: Grzelak, Edyta M;Hwang, Changhwa;Cai, Geping;Nam, Joo;Choules, Mary P;Gao, Wei;Lankin, David C;McAlpine, James B;Mulugeta, Surafel G;Napolitano, José G;Suh, Joo;Yang, Seung Hwan;Cheng, Jinhua;Lee, Hanki;Kim, Jin;Cho, Sang
• 通讯作者: Cho, Sang