Mantell Mark A, Pahil Karanbir S, Holman Karli R, Alper Joshua D, Jones Chloe M, Yao Gang, Marcaurelle Lisa A
Encoded Technologies, Molecular Modalities Discovery, GSK, Cambridge, Massachusetts 02140, United States.
J Med Chem. 2025 Sep 25;68(18):19598-19606. doi: 10.1021/acs.jmedchem.5c01885. Epub 2025 Sep 10.
DNA-encoded libraries (DELs) are used throughout small-molecule drug discovery to identify new lead compounds for protein targets. DEL hits are traditionally evaluated via off-DNA resynthesis and subsequent biological testing. This approach can be time- and resource-intensive, limiting the number of putative hits selected for follow-up. Additionally, these hits often fail to confirm off-DNA, leading to lost time and effort. Here, we introduce GSK's on-DNA hit resynthesis and binder confirmation workflow, which increases throughput and emulates the original library synthesis, thereby enabling identification of side product binders and increasing confidence in DEL hits. We utilized thermal shift, microscale thermophoresis, activity, and compound-immobilized SPR assays to develop a robust platform for the derisking of DEL hits. This is followed by a mass-spectrometry-based assay to identify specific binders in mixtures of on-DNA compounds. We share GSK's application of on-DNA binder confirmation to both evaluate and expand hits from DEL screens.
DNA编码文库（DELs）在小分子药物发现过程中被广泛用于识别针对蛋白质靶点的新先导化合物。传统上，通过非DNA重新合成及后续生物学测试来评估DEL筛选出的活性分子。这种方法可能耗费大量时间和资源，限制了后续跟进的假定活性分子数量。此外，这些活性分子在非DNA条件下往往无法得到验证，导致时间和精力的浪费。在此，我们介绍了葛兰素史克公司基于DNA的活性分子重新合成及结合物确认工作流程，该流程提高了通量并模拟了原始文库合成过程，从而能够识别副产物结合物并增强对DEL筛选出的活性分子的信心。我们利用热迁移、微量热泳动、活性及化合物固定化表面等离子体共振分析来开发一个强大的平台，用于降低DEL筛选出的活性分子的风险。接下来是基于质谱的分析，以识别DNA上化合物混合物中的特异性结合物。我们分享了葛兰素史克公司基于DNA的结合物确认方法在评估和扩展DEL筛选出的活性分子方面的应用。
