GW4869
目录号 : GC19186
GW4869是一种非竞争性中性鞘脂酶抑制剂，IC50为1微摩尔。
Cas No.:6823-69-4
Sample solution is provided at 25 µL, 10mM.
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Related Biological Data
Autophagosome accumulation is critical for efficient RABV VLP budding ACCEPTED MANUSCRIPT in cells and viral virulence in mice. (G) N2a cells infected with RABV at an MOI of 0.01 for indicated time points, and treated with GW4869 (2.5/5 μM) for 24 h.
N2a cells treated with GW4869 (2.5/5 μM) (GLPBIO) for 24 h.
Autophagy just-accepted (2024). PMID: 38566321 IF: 13.3003 -
Related Biological Data
(H–I) IPEC-J2 cells pretreated with Fumonisin B (50 µM), Myriocin (20 µM), GW4869 (10 µM), or Amitriptyline (30 µM) for 12 h and then infected with PoRV (0.1 MOI) for 6 h.
In brief, IPEC-J2 cells cultured in 6-cm dishes were infected with PoRV or treated with inhibitors (Myriocin, 20 µM; Fumonisin B1, 50 µM; Amitriptyline HCl, 30 µM; or GW4869, 10 µM) (GLPBIO) for several hours.
Journal of Virology (2024): e00064-24. PMID: 38488360 IF: 5.3999 -
Related Biological Data
Effect of exosome transplantation on klotho activation and expression. A Fluorescent report plasmid containing klotho promoter and pRL-TK were co-transfected into 293 T cells.
GW4869 (GLPBIO, Montclair, CA,USA) was used as an inhibitor of exosome biogenesis and release.
Eur J Med Res 27.1 (2022): 1-9. PMID: 35820962 IF: 4.9809 -
Related Biological Data
DM circulating exosomes exacerbate mitochondrial dysfunction of chondrocytes. A, B The mitochondrial membrane potential (△Ψ) was determined by JC-1 staining in chondrocytes.
To inhibit exosome biogenesis, the neutral sphingomyelinase-2 inhibitor, GW4869 (2 mg/kg)（GLPBIO）, was intraperitoneally injected three times a week for 12 weeks in DM-OA-GW4869 mice.
Arch Biochem Biophys (2024): 109960. PMID: 38513770 IF: 3.9003
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| 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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References: [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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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].
References:
[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.
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]。
| Cas No. | 6823-69-4 | SDF | |
| 化学名 | 3,3'-(1,4-phenylene)bis[N-[4-(4,5-dihydro-1H-imidazol-2-yl)phenyl]-dihydrochloride-2-propenamide | ||
| Canonical SMILES | O=C(NC1=CC=C(C2=NCCN2)C=C1)/C=C/C3=CC=C(/C=C/C(NC4=CC=C(C5=NCCN5)C=C4)=O)C=C3.[H]Cl.[H]Cl | ||
| 分子式 | C30H30Cl2N6O2 | 分子量 | 577.5 |
| 溶解度 | Water : < 0.1 mg/mL (insoluble); DMSO : 2 mg/mL | 储存条件 | Store at -20°C, ready to use |
| General tips | 请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。 储备液的保存方式和期限：-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。 为了提高溶解度，请将管子加热至37℃，然后在超声波浴中震荡一段时间。 |
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| Shipping Condition | 评估样品解决方案：配备蓝冰进行发货。所有其他可用尺寸：配备RT，或根据请求配备蓝冰。 | ||
| 制备储备液 | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 1.7316 mL | 8.658 mL | 17.316 mL |
| 5 mM | 0.3463 mL | 1.7316 mL | 3.4632 mL |
| 10 mM | 0.1732 mL | 0.8658 mL | 1.7316 mL |
| 第一步：请输入基本实验信息（考虑到实验过程中的损耗，建议多配一只动物的药量） | ||||||||||
| 给药剂量 | mg/kg |  |
动物平均体重 | g | |
每只动物给药体积 | ul | |
动物数量 | 只 |
| 第二步：请输入动物体内配方组成（配方适用于不溶于水的药物；不同批次药物配方比例不同，请联系GLPBIO为您提供正确的澄清溶液配方） | ||||||||||
| % DMSO % % Tween 80 % saline | ||||||||||
| 计算重置 | ||||||||||
计算结果:
工作液浓度： mg/ml；
DMSO母液配制方法： mg 药物溶于 μL DMSO溶液（母液浓度 mg/mL，
体内配方配制方法：取 μL DMSO母液，加入 μL PEG300，混匀澄清后加入μL Tween 80，混匀澄清后加入 μL saline，混匀澄清。
1. 首先保证母液是澄清的；
2.
一定要按照顺序依次将溶剂加入，进行下一步操作之前必须保证上一步操作得到的是澄清的溶液，可采用涡旋、超声或水浴加热等物理方法助溶。
3. 以上所有助溶剂都可在 GlpBio 网站选购。
Blockade of exosome generation with GW4869 dampens the sepsis-induced inflammation and cardiac dysfunction
Biochim Biophys Acta2015 Nov;1852(11):2362-71.PMID: 26300484DOI: 10.1016/j.bbadis.2015.08.010
Sepsis is an infection-induced severe inflammatory disorder that leads to multiple organ failure. Amongst organs affected, myocardial depression is believed to be a major contributor to septic death. While it has been identified that large amounts of circulating pro-inflammatory cytokines are culprit for triggering cardiac dysfunction in sepsis, the underlying mechanisms remain obscure. Additionally, recent studies have shown that exosomes released from bacteria-infected macrophages are pro-inflammatory. Hence, we examined in this study whether blocking the generation of exosomes would be protective against sepsis-induced inflammatory response and cardiac dysfunction. To this end, we pre-treated RAW264.7 macrophages with GW4869, an inhibitor of exosome biogenesis/release, followed by endotoxin (LPS) challenge. In vivo, we injected wild-type (WT) mice with GW4869 for 1h prior to endotoxin treatment or cecal ligation/puncture (CLP) surgery. We observed that pre-treatment with GW4869 significantly impaired release of both exosomes and pro-inflammatory cytokines (TNF-α, IL-1β, IL-6) in RAW264.7 macrophages. At 12h after LPS treatment or CLP surgery, WT mice pre-treated 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. Together, our findings indicate that blockade of exosome generation in sepsis dampens the sepsis-triggered inflammatory response and thereby, improves cardiac function and survival.
Blockade of lncRNA-ASLNCS5088-enriched exosome generation in M2 macrophages by GW4869 dampens the effect of M2 macrophages on orchestrating fibroblast activation
FASEB J2019 Nov;33(11):12200-12212.PMID: 31373848DOI: 10.1096/fj.201901610
In hypertrophic scar (HS) formation, the type 2 immune response induces the alternatively activated macrophages (M2), which manipulate fibroblasts to differentiate into myofibroblasts with active biologic functions and proliferation. Myofibroblasts express α-smooth muscle actin (α-SMA) and synthesize and produce additional collagen type I and collagen type III, inducing HS formation. However, studies on the mechanism of M2 macrophage modulation are only based on the recognition of profibrotic factors such as TGF-β1 secreted by macrophages. The influence of exosomes from M2 macrophages on scar formation is still unknown. Both M2 macrophages and myofibroblasts highly express glutaminases (GLSs). GLS is a critical enzyme in glutaminolysis and is important for M2 macrophage and fibroblast polarization. In this study, we found that in a TGF-β1-stimulated coculture system, a long noncoding RNA (lncRNA) named lncRNA-ASLNCS5088 was enriched in M2 macrophage-derived exosomes. This lncRNA could be transferred with high efficiency to fibroblasts and acted as an endogenous sponge to adsorb microRNA-200c-3p, resulting in increased GLS and α-SMA expression. Pretreatment with GW4869, which impairs M2 macrophage exosome synthesis, ameliorated these pathologic changes in fibroblasts in vitro. Local injection in the late scar formation period with GW4869 reduced α-SMA+ fibroblasts and alleviated the fibrosis of tissue after wound healing in vivo.-Chen, J., Zhou, R., Liang, Y., Fu, X., Wang, D., Wang, C. Blockade of lncRNA-ASLNCS5088-enriched exosome generation in M2 macrophages by GW4869 dampens the effect of M2 macrophages on orchestrating fibroblast activation.
Cancer-associated fibroblast exosomes regulate survival and proliferation of pancreatic cancer cells
Oncogene2017 Mar 30;36(13):1770-1778.PMID: 27669441DOI: 10.1038/onc.2016.353
Cancer-associated fibroblasts (CAFs) comprise the majority of the tumor bulk of pancreatic ductal adenocarcinomas (PDACs). Current efforts to eradicate these tumors focus predominantly on targeting the proliferation of rapidly growing cancer epithelial cells. We know that this is largely ineffective with resistance arising in most tumors following exposure to chemotherapy. Despite the long-standing recognition of the prominence of CAFs in PDAC, the effect of chemotherapy on CAFs and how they may contribute to drug resistance in neighboring cancer cells is not well characterized. Here, we show that CAFs exposed to chemotherapy have an active role in regulating the survival and proliferation of cancer cells. We found that CAFs are intrinsically resistant to gemcitabine, the chemotherapeutic standard of care for PDAC. Further, CAFs exposed to gemcitabine significantly increase the release of extracellular vesicles called exosomes. These exosomes increased chemoresistance-inducing factor, Snail, in recipient epithelial cells and promote proliferation and drug resistance. Finally, treatment of gemcitabine-exposed CAFs with an inhibitor of exosome release, GW4869, significantly reduces survival in co-cultured epithelial cells, signifying an important role of CAF exosomes in chemotherapeutic drug resistance. Collectively, these findings show the potential for exosome inhibitors as treatment options alongside chemotherapy for overcoming PDAC chemoresistance.