Inhibitor of neutral sphingomyelinase
GW4869 is a noncompetitive neutral sphingomyelinase inhibitor with an IC50 of 1 uM.
GW4869 (10 uM) partially inhibits TNF-induced sphingomyelin (SM) hydrolysis, and 20 uM of the compound is protected completely from the loss of SM. The addition of 10-20 uM GW4869 completely inhibits the initial accumulation of ceramide, whereas this effect is partially lost at later time points (24 h). The action of GW4869 occurs downstream of the drop in glutathione. GW4869 is able, in a dose-dependent manner, to significantly protect from cell death. These protective effects are accompanied by significant inhibition of cytochrome c release from mitochondria and caspase 9 activation, therefore localizing N-SMase activation upstream of mitochondrial dysfunction[1].
Pre-treatment with GW4869 significantly impairs release of both exosomes and pro-inflammatory cytokines (TNF-α, IL-1β, IL-6) in RAW264.7 macrophages. At 12 h after LPS treatment or CLP surgery, WT mice pretreated with GW4869 displays lower amounts of exosomes and pro-inflammatory cytokines in the serum than control PBS-injected mice. Accordingly, GW4869 treatment diminishes the sepsis-induced cardiac inflammation, attenuates myocardial depression and prolongs survival[2].
GW4869是一种非竞争性中性鞘脂酶抑制剂，IC50为1微摩尔。
GW4869是一种化合物，浓度为10微摩尔时部分抑制TNF诱导的鞘磷脂（SM）水解，而20微摩尔的化合物完全保护免受SM损失。添加10-20微摩尔的GW4869完全抑制了神经酰胺初始积累，但在后期时间点（24小时）这种效果部分丧失。 GW4869的作用发生在谷胱甘肽下降之后。 GW4869能够以剂量依赖方式显着保护细胞免受死亡。这些保护效应伴随着线粒体细胞色素c释放和半胱氨酸天冬氨酸蛋白酶9激活的显着抑制，因此将N-SMase激活定位在线粒体功能障碍上游[1]。
使用GW4869进行预处理显著影响了RAW264.7巨噬细胞释放外泌体和促炎性细胞因子（TNF-α、IL-1β、IL-6）。在LPS处理或CLP手术后12小时，与对照组PBS注射的小鼠相比，接受GW4869预处理的WT小鼠血清中外泌体和促炎性细胞因子数量较低。因此，GW4869治疗减轻了败血症引起的心脏炎症反应，缓解了心肌抑制，并延长了生存时间[2]。
参考文献:
[1]. Luberto C, et al. Inhibition of tumor necrosis factor-induced cell death in MCF7 by a novel inhibitor of neutralsphingomyelinase. J Biol Chem. 2002 Oct 25;277(43):41128-39.
[2]. Essandoh K, et al. Blockade of exosome generation with GW4869 dampens the sepsis-induced inflammation and cardiac dysfunction. Biochim Biophys Acta. 2015 Nov;1852(11):2362-71.
[3]. Nakamura H, et al. Sphingomyelin Regulates the Activity of Secretory Phospholipase A2 in the Plasma Membrane. J Cell Biochem. 2015 Sep;116(9):1898-907.
| Cell experiment [1]: | |
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Cell lines |
MCF7 human breast cancer cells |
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Preparation method |
GW-4869 was routinely stored at ?80 °C as a 1.5 mm stock suspension in DMSO. Right before use, the suspension was solubilized by the addition of 5% methane sulfonic acid (MSA) (2.5 μl of 5% MSA in sterile double-distilled H2O were added to 50 μl of GW4869 stock suspension; therefore, the concentration of the GW-4869 stock solution at the time of the experiments was 1.43 mm). The suspension was mixed and warmed up at 37 °C until clear. |
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Reaction Conditions |
30 min |
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Applications |
GW4869 is a noncompetitive neutral sphingomyelinase (N-SMase) inhibitor with an IC50 of 1 μM.GW4869 (10 μm) partially inhibited TNF-induced sphingomyelin (SM) hydrolysis, and 20 μm of the compound was protected completely from the loss of SM. The action of GW4869 occurred downstream of the drop in glutathione, which was shown previously to occur upstream of the activation of N-SMase. GW4869 does not interfere with other key TNF-mediated signaling effects. GW4869 was able, in a dose-dependent manner, to significantly protect from cell death as measured by nuclear condensation, caspase activation, PARP degradation, and trypan blue uptake. |
| Animal experiment [2]: | |
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Animal models |
Male wild-type C57BL/6 mice |
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Dosage form |
GW 4869, dissolved in DMSO (0.005%), was intraperitoneally (i.p.) injected at one dose of 2.5μg/g before sham or CLP surgery. |
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Applications |
Pre-treatment with GW 4869 significantly impaired release of both exosomes and pro-inflammatory cytokines (TNF-α, IL-1β, IL-6) in RAW264.7 macrophages. WT mice pretreated with GW4869 displayed lower amounts of exosomes and pro-inflammatory cytokines in the serum than control PBS-injected mice. Accordingly, GW4869 treatment diminished the sepsis-induced cardiac inflammation, attenuated myocardial depression and prolonged survival. |
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参考文献: [1]. Luberto, Chiara, et al. "Inhibition of tumor necrosis factor-induced cell death in MCF7 by a novel inhibitor of neutral sphingomyelinase." Journal of Biological Chemistry 277.43 (2002): 41128-41139. [2]. Essandoh, Kobina, et al. "Blockade of exosome generation with GW4869 dampens the sepsis-induced inflammation and cardiac dysfunction." Biochimica et Biophysica Acta (BBA)-Molecular Basis of Disease 1852.11 (2015): 2362-2371. |
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