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

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

GNE684 (20 μM; 20 hours) potently inhibits RIP1 kinase-driven cell death in multiple human and mouse cell lines [1]. GNE684 (20 μM; 0-60 minutes) disrupts TBZ (2 μM BV6, 20 ng/ml TNF, 20 μM zVAD)-induced RIP1 autophosphorylation, interactions between RIP1 and RIP3, RIP3 autophosphorylation, and phosphorylation of MLKL by RIP3 [1]. Cell Viability Assay [1] Cell Line: L929 cells, Jurkat cells, MEFs Concentration: 20 μM Incubation Time: 20 hours Result: Inhibited RIP1 kinase driven cell death effectively in several human and mouse cell lines. Western Blot Analysis [1] Cell Line: HT-29 cells, J774A.1 cells Concentration: 0 μM, 20 μM Incubation Time: 0 minute, 15 minutes, 60 minutes Result: Disrupted TBZ (2 μM BV6, 20 ng/ml TNF, 20μM zVAD)-induced RIP1 autophosphorylation, interactions between RIP1 and RIP3, RIP3 autophosphorylation, and phosphorylation of MLKL by RIP3.

GNE684 did not impact on overall survival or tumor growth in the KPP or KPR (LSL-Kras G12D/+; p16/p19 fl/wt; Trp53 R270H/wt; Pdx1-cre) PDAC models [1]. GNE684 (50mg/kg; p.o. twice daily) inhibits colitis and ileitis caused by NEMO deficiency in intestinal epithelial cells (IECs) [1]. Animal Model: Nemo fl/fl Villin.creERT2 mice (NEMO IEC-KO) [1] Dosage: 50 mg/kg Administration: Oral administration; twice daily; from days 2–6 treated with tamoxifen Result: Almost completely protected the NEMO-deficient intestines from colitis and ileitis.