DI-1548 is a potent, selective, irreversible covalent inhibitor of DCN1 (IC50 = 4.6 nM). It efficiently and selectively inhibits the neddylation of Cullin 3 at nanomolar concentrations without significantly affecting the neddylation of other Cullin proteins, such as Cullin 1, 2, 4A, 4B, and 5, and exhibits no cytotoxicity. The covalent bond formation between DI-1548 and DCN1 disrupts the DCN1-UBC12 interaction, leading to the disassembly of the neddylation complex and the subsequent inactivation of CRL3. This process results in the accumulation of NRF2 and the upregulation of its target genes, providing hepatoprotective effects in mouse models. DI-1548 is applicable in studies related to liver injury.

DI-1548 (1-1000 nM, 24 hours) effectively inhibits the neddylation of cullin 3 at concentrations as low as 1 nM in U2OS cells, showing approximately 1000 times greater potency than DI-591. In a range of cell lines, including U2OS, MDA-MB-231, KYSE70, and HCT116, DI-1548 (0.3-1000 nM, 24 hours) inhibits the neddylation of cullin 3 with selectivity over other cullin proteins by more than 1000-fold. It exhibits no cytotoxicity in U2OS, MDA-MB-231, HCT116, and KYSE70 cells at concentrations up to 1000 nM over 72 hours. At 30 nM for up to 48 hours, DI-1548 selectively and rapidly inhibits cullin 3 neddylation, without affecting cullin 1. DI-1548 (0.3-1000 nM, 1 hour) also stabilizes the intracellular DCN1 protein in a dose-dependent manner at nanomolar concentrations, with no significant effect on DCN3.

a single intraperitoneal dose of DI-1548 (25 mg/kg) can elevate NRF2 protein levels in the liver of C57BL/6 wild-type mice.