Synthesis of Base-Labile and Electrophilic Oligodeoxynucleotides

项目摘要 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分子形成共价键， 生物分子如DNA、mRNA和蛋白质。因此，它们可以用作探针 在研究领域，如化学生物学，并有可能成为一类新的治疗剂， 与基于小有机分子、肽和DNA类似物的药物相比，这些药物缺乏这样的功能。在 此外，在细胞中发现了含有碱不稳定和亲电子基团的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， 衍生物的可用性对于成功至关重要。因此，化学合成碱不稳定和亲电的 DNA类似物在健康相关研究中很重要。传统的DNA合成技术使用强碱性和 与碱不稳定和亲电子基团不相容的亲核试剂不适用于该目的。 一些报道的旨在解决该问题的方法具有严重的缺点，包括产品被有毒物质污染。 过渡金属、贵金属过度使用的高成本、UV光对DNA的损伤、复杂的后DNA 合成方法和狭窄的应用。该项目的目标是开发一种普遍适用的技术， 合成含有广泛的碱不稳定和亲电子官能团的DNA类似物。实现 目的，在本发明的实施方案中，将使用基于1，3-二噻烷-2-基-甲氧基有机官能团的保护基和连接基。 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、过渡金属或它们的组合。 金属和紫外线在整个过程。该技术不需要任何繁琐和复杂的后DNA合成 操纵也是。因此，它将在实际中用于合成含有碱基不稳定的DNA类似物， 亲电子基团在上一个资助期间，我们已证明，通过合成天然 非亲核条件下的DNA。在下一个资助期内，我们的具体目标包括评估 合成含有不同亲电基团的DNA类似物的技术，并进一步推进 技术提升到一个新的水平，使其更方便使用，并可能具有更广泛的基板范围。我们还将研究 在小分子合成的背景下，本项目中发明的保护基团。我们的长期目标是发展一个 基于潜在活性亲电子DNA类似物的新一代反义药物。成功完成本 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 研究人员与有价值的研究同样重要。本项目将帮助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研究人员。他们将学习技术，包括有机 自动化DNA合成等等。通过该项目，我们制药专业的本科生 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，这将提高他们在生物医学领域从事职业的兴趣和资格。

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.