Large‐Scale Profiling of Kinase Degradation by Using Norbornene‐Based Hydrophobic Tag (HyT) Strategy

Abstract

ABSTRACT Hydrophobic tags (HyTs) are promising bifunctional protein degraders that mimic misfolded proteins to trigger quality control‐mediated target degradation, offering key advantages over traditional heterobifunctional degraders such as proteolysis‐targeting chimeras (PROTACs). However, the scope and generality of this targeted protein degradation (TPD) strategy across the human kinome remain unexplored. In this study, we first addressed this gap by developing two general HyTs on the basis of a pan‐kinase scaffold capable of large‐scale proteome‐wide studies of kinase degradation by using quantitative chemoproteomics. We subsequently mapped the degradable kinome landscape by using a HyT‐based strategy, leading to the successful identification of 169 HyT‐degradable human kinases. Leveraging this comprehensive kinase degradome map, we next rationally designed norbornene‐based HyT degraders against ABL and AURKA, obtaining two optimized HyTs with potent degradation capabilities. We further elucidated the detailed mechanistic insight of these novel degraders. To overcome the inherent poor water solubility of HyTs, we next engineered HyT‐loaded, tumor microenvironment (TME)‐responsive self‐assembled nanoparticles (NPs), which showed improved tumor accumulation and therapeutic efficacy in vivo. With key advantages including rapid target identification and a modular NP‐based delivery platform, our work herein provides a comprehensive framework for future development of potential kinase therapeutics based on HyT degraders.