Oligodeoxynucleotide Synthesis Using Protecting Groups and a Linker Cleavable Und

使用保护基团和可切割连接体合成寡脱氧核苷酸

• 批准号: 8626130
• 负责人: Shiyue Fang
• 金额: $ 33.36万
• 依托单位: MICHIGAN TECHNOLOGICAL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-02-01 至 2017-09-19
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8626130
• 关键词: Acidity; Affinity; Aldehydes; Amides; Area; Aziridines; Biochemistry; Biology; Biomedical Research; Cell physiology; Cells; Chemicals; Chemistry; Cleaved cell; Collaborations; Column Chromatography; DNA; DNA Damage; DNA biosynthesis; DNA-Protein Interaction; Dimensions; Doctor of Philosophy; Drug usage; Education; Epoxy Compounds; Esters; Foundations; Generations; Goals; Health; High Pressure Liquid Chromatography; Learning; Link; Maleimides; Messenger RNA; Metals; Methodology; Methods; Modeling; Molecular Biology; N.I.H. Research Support; Nucleic Acids; Oligosaccharides; Organic Synthesis; Outcome; Peptides; Pharmaceutical Chemistry; Pharmaceutical Preparations; Play; Postdoctoral Fellow; Problem Solving; Procedures; Process; Proteins; RNA; Reaction; Reagent; Reporting; Research; Research Infrastructure; Research Personnel; Research Project Grants; Solid; Solutions; Students; Sulfides; Techniques; Technology; Therapeutic Agents; Training; Transition Elements; Ultraviolet Rays; analog; aptamer; base; career; chemical synthesis; combinatorial chemistry; commercialization; cost; covalent bond; drug development; gel electrophoresis; gene synthesis; hemiacetal; improved; instrument; interest; new technology; next generation; oxidation; phosphoramidite; programs; public health relevance; undergraduate student

Project summary Oligodeoxynucleotide Synthesis using Protecting Groups and a Linker Cleavable under Neutral Oxidative Conditions DNA analogs that contain latently reactive electrophilic functionalities can selectively form covalent bonds with target biomolecules such as DNA, mRNA, and protein through affinity induced reactions. As a result, they can be used as probes in research areas such as chemical biology, and have the potential to become a new class of therapeutic agents that have certain advantages over drugs based on small organic molecules and peptides. In addition, DNA derivatives that contain base-labile and electrophilic groups have been found in cells. They are results of important cellular processes and may play important cellular functions as well. Consequently, chemical synthesis of base-labile and electrophilic DNA analogs is important in health related research. Traditional DNA synthesis technologies use strongly basic and nucleophilic reagents, which are not compatible with base-labile and electrophilic groups, are not suitable for the purpose. A few reported methods intended to solve the problem have serious drawbacks including contamination of product by toxic transition metal, high cost of excessively used precious metal, damage of DNA by UV light, complicated post-DNA synthesis procedure, and narrow applications. The specific aim of this project is to develop a universally useful technology for the synthesis of DNA analogs that contain a wide range of base-labile and electrophilic functionalities. To achieve the aim, protecting groups and linkers based on the 1,3-dithian-2-yl-methoxy organic function will be employed during DNA synthesis. With these groups and linkers, the technology does not require using any strong base, nucleophile, transition metal, and UV light in the entire process. The technology does not need any tedious and complicated post-DNA synthesis manipulations either. As a result, it will be practically useful for the synthesis of DNA analogs containing base-labile and electrophilic groups. Our long-term objective is to develop a new generation of antisense drugs based on latently reactive electrophilic DNA analogs. Successful completion of this project will build the foundation for us to achieve the objective. Importantly, the new technology will be widely used by other biomedical researchers all over the world as well. The PI believes that cultivating next generation biomedical researchers is equally important as meritorious research itself. This project will help the PI to train one postdoc, two PhD students and about six undergraduate researchers in the area of nucleic acid chemistry. They will learn techniques including organic synthesis, flash column chromatography, HPLC, NMR, MS, automated DNA synthesis, and gel electrophoresis. With this project, undergraduate students majoring in our pharmaceutical chemistry, biochemistry & molecular biology, cheminformatics, and chemistry programs will have a chance to participate in NIH- supported research. Their interests in pursuing a career in biomedical research will be enhanced. Our Chemistry Department has required infrastructure and instruments for research and education. This project will help us to maintain and improve our ability to make continued contributions.

项目摘要 使用保护组和连接器的寡脱氧核苷酸合成 在中性氧化条件下可切合 包含潜在反应性亲电功能的DNA类似物可以选择形成共价键 通过亲和力诱导的反应，与靶向生物分子（例如DNA，mRNA和蛋白质）。结果，他们 可以用作化学生物学等研究领域的探针，并有可能成为新类 基于小的有机分子和肽的药物具有某些优势的治疗剂。 另外，在细胞中发现了包含基碱和亲电基团的DNA衍生物。他们 是重要的细胞过程的结果，也可能发挥重要的细胞功能。最后， 基本比例和亲电DNA类似物的化学合成在与健康相关的研究中很重要。 传统的DNA合成技术使用强碱性和亲核试剂，它们不兼容 具有碱基和亲电的基团，不适合该目的。一些报告的方法旨在 解决该问题具有严重的缺点，包括对产品污染有毒过渡金属，高 过度使用的贵金属的成本，紫外线损坏DNA，复杂的DNA合成 程序和狭窄的应用。该项目的具体目的是开发普遍有用的 用于合成DNA类似物的技术，这些DNA类似物包含多种基本和亲电的技术 功能。为了实现目标，请根据1,3-二硫代-2-基甲氧基保护组和接头 在DNA合成过程中将采用有机功能。对于这些组和链接器，技术确实 在整个过程中，不需要使用任何强大的碱，亲核器，过渡金属和紫外线。这 技术也不需要任何乏味且复杂的DNA合成操作。结果，它 实际上，将有助于合成含有碱基和亲电的DNA类似物。我们的 长期目的是开发基于潜在反应性亲电的新一代反义药物 DNA类似物。该项目的成功完成将为我们实现目标奠定基础。 重要的是，这项新技术也将被世界各地的其他生物医学研究人员广泛使用。 PI认为，培养下一代生物医学研究人员同样重要 研究本身。该项目将帮助PI培训一名博士后，两名博士学位学生和大约六个 核酸化学领域的本科研究人员。他们将学习包括有机的技术 合成，闪光柱色谱，HPLC，NMR，MS，自动DNA合成和凝胶电泳。 在这个项目中，主修我们的药物化学，生物化学和专业的本科生 分子生物学，化学信息学和化学计划将有机会参加NIH- 支持研究。他们对从事生物医学研究职业的兴趣将得到增强。我们的 化学部门需要研究和教育的基础设施和工具。这个项目 将帮助我们维持和提高我们做出持续贡献的能力。