IMB16-5 alleviates liver fibrosis by targeting MICALL2/β-catenin signaling

Liver fibrosis, which is driven by activated hepatic stellate cells (HSCs), lacks approved therapies. Here, we identify MICAL-like protein 2 (MICALL2) as a novel therapeutic target and elucidate the antifibrotic mechanism of IMB16-5, a first-in-class arylsulfonylamino-benzanilide derivative. IMB16-5 significantly attenuated liver fibrosis in carbon tetrachloride (CCl₄)-injured mice by reducing collagen deposition, inflammation, and HSC activation. Using limited proteolysis-small-molecule mapping (LiP-SMap) and molecular dynamics simulation, we discovered that IMB16-5 binds to the C-terminal domain of MICALL2. Mechanistically, MICALL2 prevents β-catenin from binding to GSK3β, thereby shielding β‑catenin from degradation and ultimately promoting Wnt signaling. IMB16-5 disrupts the MICALL2-β-catenin interaction, which suppresses β-catenin nuclear translocation. Consequently, IMB16-5 inhibits both glycolytic reprogramming and mitochondrial respiration in HSCs, promoting a quiescent phenotype. Further investigations revealed that HIF-1α acts downstream of the MICALL2/β-catenin signalingto integrate metabolic-inflammatory crosstalk. Critically, MICALL2 overexpression in vivo abolishes the therapeutic efficacy of IMB16-5, confirming target engagement. Our work establishes MICALL2 as a druggable target for liver fibrosis and identifies IMB16-5 as a promising therapeutic candidate that acts through dual metabolic suppression.