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.

项目概要 含有潜在反应性亲电子功能的 DNA 类似物可以选择性地与靶标形成共价键 通过亲和力诱导反应来检测 DNA、mRNA 和蛋白质等生物分子。因此，它们可以用作探针 在化学生物学等研究领域，有潜力成为一类新型治疗剂 与基于缺乏此类功能的有机小分子、肽和 DNA 类似物的药物相比，该药物具有优势。在 此外，在细胞中还发现了含有碱不稳定基团和亲电子基团的DNA衍生物。它们是中间体 重要的细胞过程并可能发挥重要的细胞功能。为了研究这些过程和功能， 衍生品的可用性对于成功至关重要。因此，碱不稳定和亲电子的化学合成 DNA 类似物在健康相关研究中非常重要。传统的 DNA 合成技术使用强碱性和 与碱不稳定基团和亲电子基团不相容的亲核试剂不适合该目的。 一些报道的旨在解决该问题的方法存在严重缺陷，包括产品被有毒物质污染 过渡金属、贵金属过度使用成本高、紫外线损伤DNA、复杂的DNA后处理 合成过程和狭窄的应用。该项目的目标是开发一种普遍适用的技术 含有多种碱不稳定和亲电功能的 DNA 类似物的合成。为了实现 目的，在过程中将使用基于 1,3-二噻吩-2-基-甲氧基有机官能团的保护基团和连接体 DNA合成。有了这些基团和连接基，该技术不需要使用任何强碱、亲核试剂、过渡剂 整个过程中使用金属和紫外线。该技术不需要任何繁琐复杂的DNA后合成 操纵。因此，它对于合成含有碱基不稳定的 DNA 类似物和 亲电基团。在之前的资助期间，我们已经证明通过合成天然物质可以实现该目标 DNA在非亲核条件下。在下一个资助期间，我们的具体目标包括评估项目的范围 含有不同亲电子基团的DNA类似物的合成技术，并进一步推进 将技术提升到一个新的水平，使其使用更加方便，并且潜在的底物范围更广。我们也会学习 该项目在小分子合成的背景下发明了保护基团。我们的长期目标是发展 基于潜在反应性亲电子 DNA 类似物的新一代反义药物。顺利完成本次 项目将为我们实现目标奠定基础。 PI相信培养下一代生物医学 研究人员与有价值的研究同样重要。该项目将帮助PI培养一名博士后，至少一名博士 学生和大约七名核酸化学本科生研究人员。他们将学习包括有机技术在内的技术 合成、自动化 DNA 合成等。通过这个项目，我们制药专业的本科生 化学、生物化学和分子生物学等项目将有机会参与NIH支持的项目 研究，这将提高他们在生物医学领域从事职业的兴趣和资格。