GSK717 is a cell-permeable NOD2 (nucleotide-binding oligomerization domain 2) inhibitor. GSK717 induces IL-8 secretion (IC50=400nM). GSK717 inhibits muramyl dipeptide-induced NOD2-mediated signaling and blocks NOD2 and TLR2 co-activation. GSK717 reduces pro-inflammatory cytokine release including IL-8, IL-6, IL-1β and TNFα by suppressing RIPK2/NF-κB and MAPK pathways. GSK717 can be used in studies on inflammatory bowel disease, infectious inflammation and NOD2-related immune disorders[1-4].
In vitro, A549, Huh7, U251, HEL-18 and ACE2-SK-N-SH cells infected with Zika virus were treated with 5-40μM GSK717 for 12-72h. GSK717 reduced viral titers, decreased the number of viral antigen-positive cells, upregulated ISG expression[5]. Human monocytes stimulated with MDP/TiO2 were treated with 5μM GSK717 for 24h. GSK717 inhibited MDP/TiO2-stimulated IL-1β production[6]. Human pulmonary microvascular endothelial cells were pretreated with 20μM GSK717 for 90min. GSK717 reduced IL-6, IL-8 and MCP-1 production[7].
In vivo, female C57BL/6 mice received a single intraperitoneal injection of 5mg/kg GSK717. The mice were orally gavaged with 100μg Lactobacillus rhamnosus GG extracellular vesicles every other day for 14 days and intravaginally inoculated with herpes simplex virus type 2. GSK717 abolished the inhibitory effect of Lactobacillus rhamnosus GG extracellular vesicles on herpes simplex virus type 2 infection, increased disease incidence, decreased survival rate, increased paralysis scores, elevated vaginal HSV-2 DNA copies[8]. Male C57BL/6JGpt mice on a chronic-plus-binge alcohol model were intraperitoneally injected with 10mg/kg GSK717 twice a week for 2 weeks (2 doses total) starting 2 weeks before heat-killed Lactobacillus johnsonii intervention. GSK717 blocked the ameliorative effect of heat-killed Lactobacillus johnsonii on alcohol-associated liver disease. GSK717 increased liver index. GSK717 upregulated ALT, AST and triglyceride levels[9]. Postnatal day 3 Sprague-Dawley rats with bilirubin encephalopathy received a single intraperitoneal injection of 10g/kg GSK717. GSK717 reduced hippocampal NOD2 expression, decreased pyroptosis-related components including IL-1β, GSDMD-N, Caspase-1 p20 and NLRP3[10].
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[5] Limonta D, Dyna-Dagman L, Branton W, et al. Nodosome inhibition as a novel broad-spectrum antiviral strategy against arboviruses, enteroviruses, and SARS-CoV-2. Antimicrob Agents Chemother. 2021 Aug;65(8):e00491-21.
[6] Svadlakova T, Kolackova M, Kulich P, et al. Human primary monocytes as a model for in vitro immunotoxicity testing: evaluation of the regulatory properties of TiO2 nanoparticles. Int J Nanomed. 2025;20:1171-89.
[7] DeWolf SE, Hawkes AA, Kurian SM, et al. Human pulmonary microvascular endothelial cells respond to DAMPs from injured renal tubular cells. Pulm Circ. 2024;14:e12379.
[8] Wang J, Chen H, Huang M, et al. Extracellular vesicles containing MDP derived from Lactobacillus rhamnosus GG inhibit HSV-2 infection by activating the NOD2-IFN-I signalling pathway. J Extracell Vesicles. 2025;14:e70152.
[9] Yin R, Wang T, Sun J, et al. Postbiotics From Lactobacillus Johnsonii Activates Gut Innate Immunity to Mitigate Alcohol-Associated Liver Disease. Adv Sci. 2025;12:2405781.
[10] Li J, Li S, Sun Q, et al. H3K18 lactylation-mediated nucleotide-binding oligomerization domain-2 (NOD2) expression promotes bilirubin-induced pyroptosis of astrocytes. J Neuroinflammation. 2025 Mar;22:76.