Macrocyclic compounds are an attractive class of therapeutic ligands against challenging targets, such as protein–protein interactions. However, the development of macrocycles as drugs is hindered by the lack of large combinatorial macrocyclic libraries, which are cumbersome, expensive, and time consuming to make, screen, and deconvolute. Here, we established a strategy for synthesizing and screening combinatorial libraries on a picomolar scale by using acoustic droplet ejection to combine building blocks at nanoliter volumes, which reduced the reaction volumes, reagent consumption, and synthesis time. As a proof-of-concept, we assembled a 2700-member target-focused macrocyclic library that we could subsequently assay in the same microtiter synthesis plates, saving the need for additional transfers and deconvolution schemes. We screened the library against the MDM2–p53 protein–protein interaction and generated micromolar and sub-micromolar inhibitors. Our approach based on acoustic liquid transfer provides a general strategy for the development of macrocycle ligands.

中文翻译：

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大环化合物库的皮摩尔级声液转移合成与筛选

大环化合物是一类有吸引力的治疗配体，可以对抗具有挑战性的目标，例如蛋白质-蛋白质相互作用。然而，大环化合物作为药物的发展受到缺乏大型组合大环化合物库的阻碍，这些组合大环化合物库的制造、筛选和解卷积繁琐、昂贵且耗时。在这里，我们建立了一种在皮摩尔规模上合成和筛选组合库的策略，通过使用声学液滴喷射来组合纳升体积的构建块，从而减少了反应体积、试剂消耗和合成时间。作为概念验证，我们组装了一个 2700 成员的以目标为中心的大环库，随后我们可以在相同的微量滴定合成板中对其进行分析，从而节省了对额外转移和解卷积方案的需要。我们针对 MDM2-p53 蛋白质-蛋白质相互作用筛选了文库，并生成了微摩尔和亚微摩尔抑制剂。我们基于声学液体转移的方法为大环配体的开发提供了一般策略。