Synthesis of Base-Labile and Electrophilic Oligodeoxynucleotides

碱不稳定和亲电子寡脱氧核苷酸的合成

• 批准号: 9376083
• 负责人: Shiyue Fang
• 金额: $ 42.67万
• 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 2022-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9376083
• 关键词: Acids; Affinity; Amination; Amines; Aniline; Area; Aziridines; Biochemistry; Biology; Biomedical Research; Carboxylic Acids; Cell physiology; Cells; Chemicals; Chemistry; Cleaved cell; DNA; DNA biosynthesis; Data; Doctor of Philosophy; Epoxy Compounds; Esters; Fluorides; Foundations; Funding; Generations; Goals; Health; Hydrogenation; Learning; Light; Maleimides; Messenger RNA; Metals; Methods; Molecular Biology; Nucleic Acids; Organic Chemistry; Organic Synthesis; Peptides; Pharmaceutical Chemistry; Pharmaceutical Preparations; Phenols; Play; Postdoctoral Fellow; Problem Solving; Procedures; Process; Proteins; Reaction; Reagent; Reporting; Research; Research Personnel; Research Project Grants; Research Support; Side; Structure; Students; Techniques; Technology; Therapeutic Agents; Time; Training; Transition Elements; UV induced DNA damage; Ultraviolet Rays; United States National Institutes of Health; Work; analog; base; career; chemical synthesis; cost; covalent bond; drug development; interest; monomer; new technology; next generation; oxidation; phosphoramidite; programs; reactivation from latency; small molecule; success; thioester; ultraviolet damage; undergraduate student

Project summary 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. Therefore, 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 advantages over drugs based on small organic molecules, peptides and DNA analogs that lack such functionalities. In addition, DNA derivatives that contain base-labile and electrophilic groups have been found in cells. They are intermediates of important cellular processes and may play important cellular functions. To study these processes and functions, the availability of the derivatives can be crucial for success. 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 objective 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 objective, 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. In the previous funding period, we have proven that the objective is achievable by synthesizing natural DNA under non-nucleophilic conditions. In the next funding period, our specific aims include evaluating the scope of the technology for the synthesis of DNA analogs that contain different electrophilic groups and further advancing the technology to a new level so that it is more convenient to use and potentially has broader substrate scope. We will also study the protecting groups invented in this project in the context of small molecule synthesis. Our long-term goal 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 goal. The PI believes that cultivating next generation biomedical researchers is equally important as meritorious research. This project will help the PI to train one postdoc, at least one PhD student and about seven undergraduate researchers in nucleic acid chemistry. They will learn techniques including organic synthesis, automated DNA synthesis, and more. With this project, undergraduate students majoring in our pharmaceutical chemistry, biochemistry & molecular biology, and other programs will have a chance to participate in NIH-supported research, which will enhance their interest and qualification in pursuing a career in biomedical field.

项目总结 含有潜在反应性亲电官能团的DNA类似物可以选择性地与靶标形成共价键 通过亲和诱导的反应，形成DNA、mRNA和蛋白质等生物分子。因此，它们可以用作探测器 在化学生物学等研究领域，并有可能成为一类新的治疗剂，具有 与基于缺乏此类功能的有机小分子、多肽和DNA类似物的药物相比，具有优势。在……里面 此外，在细胞中还发现了含有不稳定碱性基团和亲电基团的DNA衍生物。它们是中间体。 在重要的细胞过程中发挥重要作用，并可能发挥重要的细胞功能。为了研究这些过程和功能， 衍生品的可用性可能是成功的关键。因此，化学合成的碱不稳定性和亲电性 DNA类似物在健康相关研究中很重要。传统的DNA合成技术使用强基础和 亲核试剂与不稳定的碱性基团和亲电基团不相容，不适合用于这一目的。 一些已报道的解决问题的方法有严重的缺点，包括有毒物质污染产品 过渡金属，过度使用贵金属的高成本，紫外线对DNA的损伤，复杂的后DNA 合成程序，应用范围较窄。该项目的目标是开发一种普遍适用的技术 合成含有广泛的不稳定碱基和亲电功能的DNA类似物。要实现 目的：基于1，3-二硫-2-甲氧基有机官能团的保护基团和连接物将在 DNA合成。有了这些基团和连接物，该技术不需要使用任何强碱、亲核、过渡 金属，以及紫外光在整个过程中。这项技术不需要任何繁琐和复杂的DNA后合成 操控也是如此。因此，它将在合成含有碱基不稳定的DNA类似物和 亲电基团。在上一个资助期，我们已经证明了这一目标是可以通过合成天然的 DNA在非亲核条件下。在下一个资助期，我们的具体目标包括评估 含有不同亲电基团的DNA类似物的合成技术和进一步推进 技术达到了一个新的水平，使其使用更加方便，并潜在地具有更广阔的衬底范围。我们还将研究 本项目在小分子合成的背景下发明的保护基团。我们的长期目标是开发一种 基于潜在反应的亲电DNA类似物的新一代反义药物。成功完成这项工作 该项目将为我们实现这一目标奠定基础。国际医学会认为，培养下一代生物医学 研究人员与功勋卓著的研究同等重要。该项目将帮助PI培养一名博士后，至少一名博士后 学生和大约七名核酸化学的本科生。他们将学习包括有机技术在内的技术 合成、自动DNA合成等。有了这个项目，我们药学专业的本科生 化学、生物化学和分子生物学等专业将有机会参加由美国国立卫生研究院资助的 研究，这将提高他们在生物医学领域从事职业生涯的兴趣和资格。