| Description | GW4869 (GW69A) is a selective and non-competitive inhibitor of neutral sphingomyelinase N-SMase (IC50=1 μM). GW4869 also inhibits exosome synthesis/release and is commonly used in exosome-related studies. |
| In vitro | 方法：人乳腺癌细胞 MCF-7 用 GW4869 (10 μM) 预处理 30 min，再加入 TNF (3 nM) 孵育 6-24 h，使用 E. coli diacylglycerol kinase assay 检测神经酰胺水平。结果：GW4869 显著抑制了 TNF 诱导的神经酰胺积累。GW4869 对 N-SMase 有抑制作用。[1]方法：巨噬细胞 RAW264.7 用 GW4869 (10-20 μM) 预处理 2 h，随后用 LPS (1μg/mL) 孵育 24 h，检测 AchE 活性。结果：用 10μM GW4869 预处理巨噬细胞后，LPS 触发的外泌体生成在巨噬细胞中显著减弱，AChE的 活性降低了 22%。用 20μM GW4869 处理进一步增强了这种衰减。[2] |
| In vivo | 方法：为检测体内活性，将 GW4869 (2.5 μg/g) 腹腔注射给 C57BL/6 小鼠，1 h 后注射 LPS (25 μg/g)。结果：GW4869 预处理小鼠可以减弱 LPS 触发的血液中外泌体和促炎细胞因子的产生，从而减少心肌炎症。[2]方法：为检测体内活性，将 GW4869 (200 μL 0.3 mg/mL，2-2.5 μg/g) 腹腔注射给 5XFAD 小鼠，每两天一次，持续六周。结果：GW4869 通过阻止外泌体分泌来减少体内淀粉样斑块的形成。[3] |
| Cell experiments | GW4869 is routinely stored at ?80?°C as a 1.5 mM stock suspension in Me2SO. Right before use, the suspension is solubilized by the addition of 5% methane sulfonic acid (MSA) (2.5 μl of 5% MSA in sterile double-distilled Water are added to 50 μL of GW4869 stock suspension). Cells are treated with GW4869 for 30 min and then TNF is added in 10 μL/well. At the indicated time points, 25 μL of MTT stock solution are added to each well and incubated at 37?°C in 5% CO2 for 3 h. The cell viability is using the MTT assay[1]. |
| Target activity | SMase:1 μM |
| Synonyms | GW554869A, GW 4869, GW69A |
| molecular weight | 577.5 |
| Molecular formula | C30H30Cl2N6O2 |
| CAS | 6823-69-4 |
| Storage | Powder: -20°C for 3 years | In solvent: -80°C for 1 year |
| Solubility | DMSO: < 1mg/ml (insoluble）, Sonication is recommended. |
| References | 1. Luberto C, et al. Inhibition of tumor necrosis factor-induced cell death in MCF7 by a novel inhibitor of neutral sphingomyelinase. 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. Dinkins MB, et al. Exosome reduction in vivo is associated with lower amyloid plaque load in the 5XFAD mouse model of Alzheimer's disease. Neurobiol Aging. 2014 Aug;35(8):1792-800. 4. . Heat injured stromal cells‐derived exosomal EGFR enhances prostatic wound healing after thulium laser resection through EMT and NF‐κB signaling. The Prostate. 2019 May 24. 5. Ge M, Qiao Z, Kong Y, et al. Exosomes mediate intercellular transfer of non-autonomous tolerance to proteasome inhibitors in mixed-lineage leukemia[J]. Cancer Science. 2020. 6. Ge M, Qiao Z, Kong Y, et al. Exosomes mediate intercellular transfer of non–autonomous tolerance to proteasome inhibitors in mixed‐lineage leukemia[J]. Cancer Science. 2020, 111(4): 1279. 7. Wang X, Chen Q Z, Zan Y X, et al. . Exosomal miR‐145‐5p derived from orthohantavirus‐infected endothelial cells inhibits HTNV infection. The FASEB Journal. 2020, 34(10): 13809-13825. |
| Citations | 1. Han X, de Dieu Habimana J, Li A L, et al. Transcription factor EB-mediated mesenchymal stem cell therapy induces autophagy and alleviates spinocerebellar ataxia type 3 defects in neuronal cells model. Cell death & disease. 2022, 13(7): 1-12. 2. Ge M, Qiao Z, Kong Y, et al. Exosomes mediate intercellular transfer of non-autonomous tolerance to proteasome inhibitors in mixed-lineage leukemia. Cancer Science. 2020 3. Wang X, Chen Q Z, Zan Y X, et al. Exosomal miR‐145‐5p derived from orthohantavirus‐infected endothelial cells inhibits HTNV infection. The FASEB Journal. 2020, 34(10): 13809-13825. 4. Shi F, Deng Z, Zhou Z, et al. Heat injured stromal cells‐derived exosomal EGFR enhances prostatic wound healing after thulium laser resection through EMT and NF‐κB signaling. The Prostate. 2019 May 24 5. Han C, Xia X, Jiao S, et al. Tripartite motif containing protein 37 involves in thrombin stimulated BV-2 microglial cell apoptosis and interleukin 1β release. Biochemical and biophysical research communications. 2019, 516(4): 1252-1257. 6. Cao Z, Li P, Li Y, et al.Encapsulation of Nano‐Bortezomib in Apoptotic Stem Cell‐Derived Vesicles for the Treatment of Multiple Myeloma.Small.2023: 2301748. 7. Wang W, Sun J C, Ye P, et al.miR‐22‐3p in the rostral ventrolateral medulla promotes hypertension through inhibiting β‐arrestin‐1.The Journal of Physiology.2023 |