Oligodeoxynucleotide Synthesis Using Protecting Groups and a Linker Cleavable Und

项目总结 利用保护基团和连接物合成寡核苷酸 在中性氧化条件下可解离 含有潜在反应性亲电官能团的DNA类似物可以选择性地形成共价键 通过亲和诱导反应与靶生物分子如DNA、mRNA和蛋白质发生反应。因此，他们 可用作化学生物学等研究领域的探针，并有可能成为一种新的类别 与以有机小分子和多肽为基础的药物相比，具有一定优势的治疗剂。 此外，在细胞中还发现了含有不稳定碱性基团和亲电性基团的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。因此， 碱基不稳定和亲电的DNA类似物的化学合成在与健康相关的研究中非常重要。 传统的DNA合成技术使用的是强碱性和亲核性试剂，它们是不相容的 具有不稳定碱性和亲电性的基团，不适合该用途。一些已报道的方法旨在 解决问题有严重的缺点，包括产品受到有毒过渡金属的污染， 过度使用贵金属的代价，紫外线对DNA的破坏，复杂的DNA后合成 程序，和狭窄的应用程序。这个项目的具体目标是开发一种普遍适用的 合成含有多种碱基不稳定和亲电性DNA类似物的技术 功能。为了达到目的，基于1，3-二硫-2-甲氧基保护基团和连接物 在DNA合成过程中将使用有机功能。有了这些群和链接器，这项技术就做到了 在整个过程中不需要使用任何强碱、亲核、过渡金属和紫外光。这个 技术也不需要任何乏味和复杂的DNA合成后操作。因此，它 对于合成含有不稳定碱性基团和亲电基团的DNA类似物具有实际应用价值。我们的 长期目标是开发基于潜伏性亲电反应的新一代反义药物 DNA类似物。这一项目的顺利完成将为我们实现这一目标奠定基础。 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。 国际医学会认为，培养下一代生物医学研究人员与功勋同等重要 研究本身。该项目将帮助PI培训一名博士后、两名博士生和大约六名博士后 核酸化学领域的本科生研究人员。他们将学习包括有机技术在内的技术 合成、闪光柱层析、高效液相、核磁共振、质谱学、DNA自动合成和凝胶电泳。 通过这个项目，我们的药物化学、生物化学和化学专业的本科生 分子生物学、化学信息学和化学项目将有机会参加NIH- 受资助的研究。他们从事生物医学研究的兴趣将会增强。我们的 化学系需要研究和教育所需的基础设施和仪器。这个项目 将有助于我们保持和提高继续作出贡献的能力。

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.