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.

项目摘要 包含潜在反应性亲电功能的DNA类似物可以选择性地形成共价键 通过亲和力诱导的反应等生物分子，例如DNA，mRNA和蛋白质。因此，它们可以用作探针 在化学生物学等研究领域，有可能成为具有新的治疗剂 基于缺乏此类功能的小有机分子，肽和DNA类似物的药物优于药物的优点。在 此外，在细胞中发现了含有碱基和亲电的DNA衍生物。他们是中间体 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， 衍生物的可用性对于成功至关重要。因此，碱基和亲电的化学合成 DNA类似物在健康相关研究中很重要。传统的DNA合成技术使用强大的基础和 与碱性和亲电的基团不兼容的亲核试剂不适合该目的。 旨在解决该问题的一些报告的方法具有严重的缺点，包括对产品污染的污染 过渡金属，过度使用的贵金属的高成本，紫外线损坏DNA，复杂的DNA 合成程序和狭窄的应用。该项目的目的是开发一种普遍有用的技术 DNA类似物的合成，包含多种基本和亲电的功能。实现 目的，将在1,3-DITHIAN-2-甲氧基有机功能中保护组和接头。 DNA合成。对于这些组和接头，该技术不需要使用任何强大的基础，亲核，过渡 金属和紫外线在整个过程中。该技术不需要任何乏味且复杂的DNA合成 操纵。结果，它实际上对于合成包含碱基的DNA类似物和 亲电基团。在上一个资金期间，我们已经证明，通过综合自然可以实现目标 DNA在非核粉条件下。在下一个资金期间，我们的具体目标包括评估范围 合成包含不同亲电基团的DNA类似物的技术，并进一步推进 技术达到新的水平，因此使用更方便，并且可能具有更广泛的底物范围。我们还将学习 该项目在小分子合成的背景下发明的保护组。我们的长期目标是开发 新一代的反义药物基于潜在反应性电力DNA类似物。成功完成 项目将为我们实现目标奠定基础。 PI认为培养下一代生物医学 研究人员与功绩性研究同样重要。该项目将帮助PI培训一个博士后，至少一位博士学位 学生和大约七名核酸化学研究人员。他们将学习包括有机的技术 合成，自动DNA合成等等。有了这个项目，我们的药品专业的本科生 化学，生物化学和分子生物学以及其他程序将有机会参加NIH支持的 研究将增强他们从事生物医学领域职业的兴趣和资格。