GNE684 is a potent inhibitor of receptor interacting protein 1 (RIP1) with average Ki app values of 21 nM for human, 189 nM for mouse, and 691 nM for rat RIP1, respectively [1].

RIP1 (mouse):189 nM , RIP1 (human):21 nM , RIP1 (rat):691 nM

GNE684, at a concentration of 20 μM for 20 hours, effectively inhibits RIP1 kinase-driven cell death across various human and mouse cell lines, as evidenced by cell viability assays in L929, Jurkat, and MEF cells. Furthermore, within a timeframe of 0 to 60 minutes, GNE684 disrupts TBZ-induced processes in HT-29 and J774A.1 cells, including RIP1 autophosphorylation, the interaction between RIP1 and RIP3, RIP3 autophosphorylation, and RIP3-mediated phosphorylation of MLKL, at the same concentration without altering conditions. These findings are supported by Western blot analysis, demonstrating GNE684's potent inhibitory effects on key mechanisms contributing to cell death and signaling pathways.

GNE684 did not affect overall survival or tumor growth in the KPP or KPR (LSL-Kras G12D/+; p16/p19 fl/wt; Trp53 R270H/wt; Pdx1-cre) PDAC models. However, GNE684 (50 mg/kg; p.o. twice daily) inhibits colitis and ileitis caused by NEMO deficiency in intestinal epithelial cells (IECs) [1]. In the Nemo fl/fl Villin.creERT2 mice (NEMO IEC-KO) model, oral administration of 50 mg/kg twice daily from days 2–6 with tamoxifen almost completely protected the NEMO-deficient intestines from colitis and ileitis.