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.

项目概要 使用保护基团和接头合成寡脱氧核苷酸 在中性氧化条件下可裂解 含有潜在反应性亲电子功能的 DNA 类似物可以选择性地形成共价键 通过亲和力诱导反应与目标生物分子（例如 DNA、mRNA 和蛋白质）发生反应。结果，他们 可作为化学生物学等研究领域的探针，有潜力成为新一类 与基于有机小分子和肽的药物相比具有某些优势的治疗剂。 此外，在细胞中还发现了含有碱不稳定基团和亲电基团的DNA衍生物。他们 是重要细胞过程的结果，也可能发挥重要的细胞功能。最后， 碱不稳定和亲电子 DNA 类似物的化学合成在健康相关研究中非常重要。 传统DNA合成技术使用强碱性和亲核试剂，两者不兼容 具有碱不稳定和亲电子基团，不适合该目的。一些报道的方法旨在 解决该问题具有严重的缺点，包括有毒过渡金属污染产品、高 过度使用贵金属的成本、紫外线对 DNA 的损伤、复杂的 DNA 后合成 程序和狭窄的应用程序。该项目的具体目标是开发一种普遍有用的 用于合成含有多种碱不稳定和亲电子的 DNA 类似物的技术 功能。为了实现这一目标，基于 1,3-二硫杂环己烷-2-基-甲氧基的保护基团和连接基 DNA合成过程中将使用有机功能。有了这些基团和连接体，该技术就可以 整个过程不需要使用任何强碱、亲核试剂、过渡金属和紫外光。这 技术也不需要任何繁琐和复杂的 DNA 合成后操作。结果，它 对于含有碱不稳定基团和亲电基团的 DNA 类似物的合成将具有实际用途。我们的 长期目标是开发基于潜在反应性亲电子的新一代反义药物 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与立功同等重要 研究本身。该项目将帮助PI培养一名博士后、两名博士生和约六名 核酸化学领域的本科研究人员。他们将学习包括有机技术在内的技术 合成、快速柱色谱、HPLC、NMR、MS、自动 DNA 合成和凝胶电泳。 通过这个项目，我们药物化学、生物化学和 分子生物学、化学信息学和化学项目将有机会参加 NIH- 支持的研究。他们从事生物医学研究事业的兴趣将会增强。我们的 化学系拥有研究和教育所需的基础设施和仪器。这个项目 将帮助我们保持和提高持续贡献的能力。