Hangke Ma, Peng Chen, Siyue Chen, Han Zhang, Fanming Zeng, Zhijun Han, Li Chen, Ayun Luo, Huanqing Zhang, Zhaomei Sun, Kehan Zhou, Lijun Xue, Kexin Yang, Yun Jin Hu

ACS Medicinal Chemistry Letters

DOI: 10.1021/acsmedchemlett.6c00278

Abstract

High-throughput screening (HTS) and DNA-encoded library (DEL) selection are cornerstones of drug discovery but suffer from high operational infrastructure requirements or constraints to affinity-only selections. Herein we report a robust on-DNA compound screening method that bridges this gap, enabling direct functional evaluation of DNA-linked small molecules while bypassing plate-based or selection-wash limitations. To validate this platform, a focused collection of trisubstituted benzamides was synthesized and screened directly on-DNA against p38α MAPK. This methodology rapidly prioritized advanced structures, culminating in the seamless identification of compound I-13. Upon off-DNA synthesis, I-13 was confirmed as a potent p38α inhibitor (IC50 = 65 nM), demonstrating 4-fold selectivity over p38β and excellent selectivity over p38γ and p38δ (>461 fold). While achieving reasonable p38α/β isoform selectivity remains a subject for future optimization, this platform offers a powerful, low-barrier alternative to traditional HTS and DEL for rapid, functional hit-to-lead discovery.