Merkin Institute of Transformative Technologies in Healthcare, Broad Institute of MIT and Harvard, Cambridge, MA, USA.
Howard Hughes Medical Institute, Harvard University, Cambridge, MA, USA.
Nat Chem. 2018 Jul;10(7):704-714. doi: 10.1038/s41557-018-0033-8. Epub 2018 Apr 2.
DNA-encoded libraries have emerged as a widely used resource for the discovery of bioactive small molecules, and offer substantial advantages compared with conventional small-molecule libraries. Here, we have developed and streamlined multiple fundamental aspects of DNA-encoded and DNA-templated library synthesis methodology, including computational identification and experimental validation of a 20 × 20 × 20 × 80 set of orthogonal codons, chemical and computational tools for enhancing the structural diversity and drug-likeness of library members, a highly efficient polymerase-mediated template library assembly strategy, and library isolation and purification methods. We have integrated these improved methods to produce a second-generation DNA-templated library of 256,000 small-molecule macrocycles with improved drug-like physical properties. In vitro selection of this library for insulin-degrading enzyme affinity resulted in novel insulin-degrading enzyme inhibitors, including one of unusual potency and novel macrocycle stereochemistry (IC = 40 nM). Collectively, these developments enable DNA-templated small-molecule libraries to serve as more powerful, accessible, streamlined and cost-effective tools for bioactive small-molecule discovery.
DNA 编码文库已成为发现生物活性小分子的广泛应用资源，与传统小分子文库相比具有显著优势。在这里，我们开发并简化了 DNA 编码和 DNA 模板库合成方法的多个基本方面，包括计算鉴定和实验验证 20×20×20×80 组正交密码子，用于增强库成员结构多样性和类药性的化学和计算工具，高效的聚合酶介导的模板库组装策略，以及文库分离和纯化方法。我们整合了这些改进的方法，生成了具有改进的类药性物理性质的第二代 256000 个小分子大环的 DNA 模板库。该文库针对胰岛素降解酶亲和力的体外选择产生了新型胰岛素降解酶抑制剂，包括一种具有不寻常效力和新型大环立体化学（IC=40nM）的抑制剂。总的来说，这些发展使 DNA 模板小分子文库成为更强大、更易于获取、更精简和更具成本效益的生物活性小分子发现工具。
