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

这个研究项目的第一个目标是开发一种化学稳定的遗传标签。它将比目前的DNA条形码策略提供更广泛的文库设计方法，并比hexT方法更有效地访问编码文库。新的标记策略依赖于文献中已知的胸腺嘧啶-、胞嘧啶-和7-去氮杂腺嘌呤核碱基对Brønsted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将与聚乙二醇聚合物结合，在选择的有机溶剂中溶解该分子。这一策略将导致一种新的头套设计，由于增强的DNA稳定性和在有机溶剂中进行反应的选择，使反应范围更大。目前正在进行的对受控孔玻璃（CPG）固相的dna化学研究将作为后备计划。这两种方法将在产品收率，DNA稳定性和操作简易性方面进行比较。与多特蒙德工业大学的Norbert Kockmann教授合作，采取措施将编码化合物库合成与实验室自动化和高通量实验相结合。优化活动的化合物合成将执行与机器人支持试剂的剂量和分配。开发的合成机器人平台将用于进一步开发dna编码化学，并有机会并行化成功的合成路线。对于基于固相的方法，设备和处理策略将被设计为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。化学稳定代码合成各种dna标记目标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-和ola -化学转化为编码格式（苏黎世联邦理工学院）。一种文库合成策略将用于合成一个概念验证的编码文库，目标是泛素E3连接酶的凋亡抑制蛋白（IAP）家族。

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.