Mixed lineage kinase domain-like (MLKL) is the terminal protein in the pro-inflammatory necroptotic cell death program. RIPK3-mediated phosphorylation is thought to initiate MLKL oligomerization, membrane translocation and membrane disruption[3]. MLKL promotes vascular inflammation by regulating the expression of adhesion molecules ICAM1, VCAM1, and E-selectin in endothelial cells (EC). MLKL deficiency suppresses the expression of these adhesion molecules, thereby reducing EC-leukocyte interaction in vitro and in vivo[4]. MLKL-induced membrane depolarization and cell death exhibit a positive correlation to its channel activity. The Mg(2+)-preferred permeability and five transmembrane segment topology distinguish MLKL from previously identified Mg(2+)-permeable channels and thus establish MLKL as a novel class of cation channels[5].Mice deficient in MLKL (Mlkl-/-) had reduced neuronal death (24 h) and BBB permeability at 24 h but not 30d, and improved post-injury rotarod performance vs. WT[6]. MLKL activation is sufficient to induce the processing and release of bioactive IL-1β. MLKL activation triggers potassium efflux and assembly of the NLRP3 inflammasome, which is required for the processing and activity of IL-1β released during necroptosis. Notably, MLKL activation also causes cell membrane disruption, which allows efficient release of IL-1β independently of the recently described pyroptotic effector gasdermin-D[7]. GW806742X, an ATP mimetic and a potent MLKL inhibitor, binds the MLKL pseudokinase domain with a Kd of 9.3 μM. GW806742X has activity against VEGFR2 (IC50=2 nM). GW806742X retards MLKL membrane translocation and inhibits necroptosis[8].
GW806742X 细胞实验
Cell Line
Concentration
Treated Time
Description
References
MEF cells
1 µM
1 hour
GW806742X, as an MLKL inhibitor, was used to study its role in complement-dependent cytotoxicity. Results showed that GW806742X failed to inhibit CDC in Bid KO MEFs.
Front Immunol. 2018 Feb 23;9:306
murine bone marrow-derived macrophages
1 μM
6 hours
inhibited MSU crystal-induced necrosis
Acta Pharmacol Sin. 2022 May;43(5):1324-1336
K-562 leukemia cells
1 µM
72 hours
GW806742X significantly reduced AURKA and AURKB expression and induced apoptosis and G2/M phase cell cycle arrest.
Sci Rep. 2020 Dec 4;10(1):21272
RAW264.7 cells
2 μM
6 hours
GW806742X significantly reduced LDH release and the proportion of necrotic cells in E. faecalis-infected RAW264.7 cells and decreased the expression of inflammatory cytokines.
Microbiol Spectr. 2022 Aug 31;10(4):e0104522
U937 monocytes
1 µM
24 hours
In high glucose conditions, GW806742X, as an MLKL inhibitor, prevented cell death.
Int J Mol Sci. 2023 May 11;24(10):8609
AsPC-1 cells
1 µM
48 hours
Inhibited MLKL-driven necroptosis, reduced levels of necroptosis markers p-MLKL and MLKL oligomers
Nat Commun. 2024 Jul 18;15(1):6043
PANC-1 cells
1 µM
48 hours
Inhibited MLKL-driven necroptosis, reversed growth arrest and increased necroptosis level (p-MLKL level) in T1M1 PDOs
Nat Commun. 2024 Jul 18;15(1):6043
SW620 cells
1 μM
1.5 hours
To evaluate the inhibitory effect of GW806742X on EBI-induced necroptosis in SW620 cells
Front Pharmacol. 2023 Jun 16;14:1219362
GW806742X 动物实验
Species
Animal Model
Administration
Dosage
Frequency
Description
References
C57BL/6 mice
Orthotopic xenograft model and liver metastasis model
Intravenous injection
100 µM in 50 µl
3 times per week for 4 weeks
GW treatment reduced the incidence of liver metastasis without affecting primary tumor lesions, and the combination regimen with anti-CD47 showed the best tumor control and survival improvement
Nat Commun. 2024 Jul 18;15(1):6043
Mice
S. marcescens pneumonia model
Intraperitoneal injection
100 μl of 100 μM
Every 4h for the first 12h post-infection
Reduced bacterial burden in the lungs and protection of alveolar macrophages
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