Development of strategies for high throughput DNA-encoded synthesis of heterocycles, and application to targeting inhibitors of apoptosis proteins (IAPs) as model systems

开发高通量 DNA 编码杂环合成策略，并将其应用于靶向凋亡蛋白 (IAP) 抑制剂作为模型系统

• 批准号: 439638519
• 负责人: Professor Dr. Andreas Brunschweiger
• 金额: --
• 依托单位: Laboratorium für Organische Chemie
• 依托单位国家: 德国
• 项目类别: Research Grants
• 资助国家: 德国
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/439638519?language=en
• 关键词: Development; strategies; throughput; DNA; encoded

The first goal of this research project is the development of a chemically stabilized genetic tag. It shall facilitate a broader scope of methods for designing libraries than the headpiece DNA bar-coding strategy, and a more efficient access to encoded libraries than the hexT approach. The new tagging strategy relies on the literature-known chemical stability of thymine-, cytosine-, and 7-desazaadenine nucleobases to Brønsted and Lewis acids.The second goal is the development of an approach to performing chemistry on chemically stabilized DNA barcodes in organic solvents. A chemically stabilized DNA will be conjugated to a polyethylene glycol polymer to solubilize this molecule in an organic solvent of choice. This strategy shall lead to a new headpiece design that enables a greater scope of reactions due to enhanced DNA stability and the option to perform reactions in organic solvents. Ongoing efforts for on-DNA chemistry on controlled pore glass (CPG) solid phase as previously published, will serve as a back-up plan. The two approaches will be compared in terms of product yields, DNA stability, and operational ease.A collaboration with Prof. Norbert Kockmann, TU Dortmund, takes steps to combine encoded compound library synthesis with laboratory automation and high-throughput experimentation. Optimization campaigns for compound synthesis will be performed with robotic support for dosing and dispension of reagents. The developed synthesis robot platform will be used to further develop DNA-coded chemistry with the opportunity of parallelization of successful synthesis routes. For the solid phase-based approaches, equipment and handling strategies will be designed that allow for repeating synthesis and washing operations in a single well plate. This will open up novel pathways for robust and high-yielding chemical syntheses in encoded library production.The suitability of the chemically stabilized code to synthesize a variety of DNA-tagged target molecules will be tested with methods that were compatible with the TiDEC strategy and through an ongoing collaboration with Prof. Jeffrey Bode to translate so-called SnAP- and OLA-chemistry to an encoded format (ETH Zurich).One library synthesis strategy will then be used to synthesize a proof-of-concept encoded library targeting the Inhibitor of Apoptosis Protein (IAP) family of ubiquitin E3 ligases.

这项研究项目的第一个目标是开发一种化学稳定的基因标签。它将促进比头戴式DNA条形码策略更广泛的库设计方法，并比Hext方法更有效地访问编码库。新的标记策略依赖于文献中已知的胸腺嘧啶、胞嘧啶和7-去氮腺嘌呤碱基对Brnsted酸和Lewis酸的化学稳定性。第二个目标是开发一种在有机溶剂中对化学稳定的DNA条形码进行化学处理的方法。化学稳定的DNA将连接到聚乙二醇聚合物上，使该分子在所选的有机溶剂中增溶。这一策略将导致一种新的头盔设计，由于增强了DNA的稳定性，并可以选择在有机溶剂中进行反应，因此可以实现更大范围的反应。正在进行的关于可控孔玻璃(CPG)固相上的DNA化学的努力，将作为后备计划。这两种方法将在产品产量、DNA稳定性和操作简易性方面进行比较。与诺伯特·科克曼教授合作，TU Dortmund采取措施将编码化合物文库合成与实验室自动化和高通量实验相结合。化合物合成的优化活动将在机器人的支持下进行，以进行试剂的配料和拆分。开发的合成机器人平台将用于进一步发展DNA编码化学，并有机会将成功的合成路线并行化。对于以固相为基础的方法，将设计设备和处理策略，允许在单个孔板中重复合成和洗涤操作。这将为编码库生产中稳健和高产的化学合成开辟新的途径。化学稳定的代码用于合成各种DNA标记的靶分子的适用性将通过与TiDEC策略兼容的方法进行测试，并通过与Jeffrey Bode教授的持续合作将所谓的Snap-and Ola化学转换为编码格式(ETH Zurich)。然后，一个文库合成策略将用于合成针对泛素E3连接酶的凋亡蛋白抑制物(IAP)家族的概念验证编码库。