Discovery of First-In-Class Potent and Selective Tropomyosin Receptor Kinase Degraders

We report compounds 5 (CG416) and 6 (CG428) as two first-in-class tropomyosin receptor kinase (Trk) degraders that target the intracellular kinase domain of Trk. Degraders 5 and 6 reduced levels of the tropomyosin 3 (TPM3)-TRKA fusion protein in KM12 colorectal carcinoma cells and inhibited downstream PLCγ1 signaling at sub-nanomolar concentrations. Both degraders also degraded human wild-type TrkA with similar potency. Interestingly, both degraders, especially 6, showed selectivity for the degradation of endogenous TPM3-TRKA over ectopically expressed ATP/GTP binding protein-like 4 (AGBL4)-TRKB or ETS variant transcription factor 6 (ETV6)-TRKC fusion proteins in KM12 cells. Global proteomic profiling assays demonstrated that 5 is highly selective for the intended target. TPM3-TRKA protein degradation induced by 5 and 6 was further confirmed to be mediated through Cereblon and the ubiquitin-proteasome system. Compared with the parental Trk kinase inhibitor, both degraders exhibited higher potency for inhibiting growth of KM12 cells. Moreover, both 5 and 6 showed good plasma exposure levels in mice. Therefore, 5 and 6 are valuable chemical tool compounds for investigating the in vivo function of Trk fusion during tumorigenesis. Our study also paves the way for pharmacological degradation of Trk.