Synthesis and Structural Optimization of ATG4B Inhibitors for the Attenuation of Treatment-Induced Autophagy in Glioblastoma

Glioblastoma, a prevalent malignant CNS tumor, presents a therapeutic challenge because of resistance to standard treatments including radiation therapy and temozolomide. Both modalities induce autophagy, paradoxically promoting tumor survival. The cysteine protease ATG4B is implicated in this cellular process, highlighting the enzyme as a viable therapeutic target for glioblastoma. We have developed streamlined syntheses for ATG4B inhibitor NSC185058, its derivatives, and fluorogenic ATG4B substrate pim-FG-PABA-AMC. We leveraged these findings to rapidly identify novel compound MJO445, which demonstrates markedly greater potency biochemically and in cells.