Institute of Chemical Sciences and Engineering, Ecole Polytechnique Fédérale de Lausanne (EPFL), CH-1015, Lausanne, Switzerland.
Biomolecular Screening Facility, Ecole Polytechnique Fédérale de Lausanne (EPFL), Lausanne, Switzerland.
Nat Commun. 2022 Jul 2;13(1):3823. doi: 10.1038/s41467-022-31428-8.
Macrocycles have excellent potential as therapeutics due to their ability to bind challenging targets. However, generating macrocycles against new targets is hindered by a lack of large macrocycle libraries for high-throughput screening. To overcome this, we herein established a combinatorial approach by tethering a myriad of chemical fragments to peripheral groups of structurally diverse macrocyclic scaffolds in a combinatorial fashion, all at a picomole scale in nanoliter volumes using acoustic droplet ejection technology. In a proof-of-concept, we generate a target-tailored library of 19,968 macrocycles by conjugating 104 carboxylic-acid fragments to 192 macrocyclic scaffolds. The high reaction efficiency and small number of side products of the acylation reactions allowed direct assay without purification and thus a large throughput. In screens, we identify nanomolar inhibitors against thrombin (K = 44 ± 1 nM) and the MDM2:p53 protein-protein interaction (K MDM2 = 43 ± 18 nM). The increased efficiency of macrocycle synthesis and screening and general applicability of this approach unlocks possibilities for generating leads against any protein target.
大环化合物因其能够结合具有挑战性的靶标而具有巨大的治疗潜力。然而，由于缺乏用于高通量筛选的大型大环化合物库，针对新靶标的大环化合物的生成受到了阻碍。为了克服这一问题，我们通过将无数化学片段以组合方式连接到结构多样的大环支架的外围基团上来建立一种组合方法，所有这些都在纳升体积的皮摩尔规模下使用声控液滴喷射技术进行。在概念验证中，我们通过将 104 个羧酸片段连接到 192 个大环支架上来生成一个针对目标的 19968 个大环化合物库。酰化反应的高反应效率和少量副产物使得无需纯化即可直接进行测定，从而实现了高通量。在筛选中，我们鉴定出了针对凝血酶（K=44±1nM）和 MDM2：p53 蛋白-蛋白相互作用（KMDM2=43±18nM）的纳摩尔抑制剂。这种大环化合物合成和筛选效率的提高以及该方法的通用性为针对任何蛋白质靶标生成先导化合物提供了可能性。
