GW806742X, an ATP mimetic and a potent MLKL (Mixed Lineage Kinase Domain-Like protein) inhibitor, binds the MLKL pseudokinase domain with a KD of 9.3μM[1]. GW806742X binds to the ATP pocket of MLKL at the pseudokinase domain by forming two hydrogen bonds, thereby inhibiting necroptosis without impacting apoptosis[2]. GW806742X has been widely used in preclinical models to alleviate acute lung injury in mice and reduce airway inflammation[3].
In vitro, GW806742X treatment for 48 hours inhibited the viability of MDA-MB-231, MDA-MB-468, and MCF 10A cells, with IC50 values of 4.024µM, 3.67µM, and 18.7µM, respectively[4]. 1µM of GW806742X pretreatment for 30 minutes suppressed the phosphorylation of MLKL in L929 cells and reduced autophagy of the cells[5]. Pretreatment with 5µM GW806742X for 1 hour reduced monosodium urate (MSU) crystal-induced necrosis of mouse macrophages and prevented inflammasome activation[6]. Treatment with 0.1µM GW806742X for 24 hours inhibited the maturation and CXCL10 secretion in mouse bone marrow-derived dendritic cells (BMDCs)[7].
In vivo, a single intraperitoneal injection of 100µl of 100µM GW806742X reduced lung bacterial burden and alleviated pathological features associated with Serratia marcescens hemorrhagic pneumonia in mice after 24 hours[8]. A single intraperitoneal injection of GW806742X at a dose of 2mg/kg alleviated cecal ligation and puncture (CLP)-induced multiple organ dysfunction and systemic inflammation and improved survival in mice after 24 hours[9].
References:[1] Hildebrand J M, Tanzer M C, Lucet I S, et al. Activation of the pseudokinase MLKL unleashes the four-helix bundle domain to induce membrane localization and necroptotic cell death[J]. Proceedings of the National Academy of Sciences, 2014, 111(42): 15072-15077.
[2] Zhou Y, Cai Z, Zhai Y, et al. Necroptosis inhibitors: mechanisms of action and therapeutic potential[J]. Apoptosis, 2024, 29(1): 22-44.
[3] Kang Z, Xuan N, Zhou Q, et al. Targeting alveolar epithelial cells with lipid micelle-encapsulated necroptosis inhibitors to alleviate acute lung injury[J]. Communications Biology, 2025, 8(1): 573.
[4] Liu W, Xu J, Chen L, et al. Discovery of a novel BCL-2 inhibitor GW806742X for the treatment of TNBC[J]. Biochemical Pharmacology, 2025, 237: 116925.
[5] Zhan Q, Jeon J, Li Y, et al. CAMK2/CaMKII activates MLKL in short-term starvation to facilitate autophagic flux[J]. Autophagy, 2022, 18(4): 726-744.
[6] Zhong C, Zeng B, Qiu J, et al. Gout-associated monosodium urate crystal-induced necrosis is independent of NLRP3 activity but can be suppressed by combined inhibitors for multiple signaling pathways[J]. Acta Pharmacologica Sinica, 2022, 43(5): 1324-1336.
[7] Jensen T S, Laursen M F, Schort L, et al. STING and Nonnecroptotic MLKL‐Mediated Mechanisms Improve Dendritic Cell Maturation and Killing of Cancer Cells[J]. European Journal of Immunology, 2025, 55(9): e70044.
[8] González-Juarbe N, Gilley R P, Hinojosa C A, et al. Pore-forming toxins induce macrophage necroptosis during acute bacterial pneumonia[J]. PLoS pathogens, 2015, 11(12): e1005337.
[9] Zhang X, Wu J, Liu Q, et al. RIPK3–MLKL necroptotic signalling amplifies STING pathway and exacerbates lethal sepsis[J]. Clinical and translational medicine, 2023, 13(7): e1334.