Formaat : 10mg
Synonyms: GW69A, GW554869A, GW 4869
| 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.Usually prepared as a suspension for experiments. |
| Targets & IC50 | SMase:1 μM |
| In vitro | METHODS: Human breast cancer cells MCF-7 were pretreated with GW4869 (10 μM) for 30 min, then incubated with TNF (3 nM) for 6-24 h. Ceramide levels were detected by E. coli diacylglycerol kinase assay. RESULTS: GW4869 significantly inhibited TNF-induced ceramide accumulation, and GW4869 inhibited N-SMase. [1] METHODS: Macrophage RAW264.7 was pretreated with GW4869 (10-20 μM) for 2 h, and then incubated with LPS (1 μg/mL) for 24 h. AchE activity was measured. RESULTS: After pretreatment of macrophages with 10 μM GW4869, LPS-triggered exocytosis was significantly attenuated in macrophages, and the activity of AChE was reduced by 22%. Treatment with 20 μM GW4869 further enhanced this attenuation. [2] |
| In vivo | METHODS: To detect in vivo activity, GW4869 (2.5 μg/g) was injected intraperitoneally into C57BL/6 mice, and LPS (25 μg/g) was injected 1 h later. RESULTS: Pretreatment of mice with GW4869 attenuated LPS-triggered production of exosomes and pro-inflammatory cytokines in the blood, thereby reducing myocardial inflammation. [2] METHODS: To assay in vivo activity, GW4869 (200 μL of 0.3 mg/mL, 2-2.5 μg/g) was administered intraperitoneally to 5XFAD mice every two days for six weeks. RESULTS: GW4869 reduced amyloid plaque formation in vivo by blocking exosome secretion. [3] |
| Synonyms | GW69A, GW554869A, GW 4869 |
| Kinase Assay | B. cereu sphosphatidylcholine-PLC is incubated in the presence or absence of 10 μM GW4869 in a reaction mixture containing 100 mM Tris, pH 7.2, 25% glycerol, 20 mM p-nitrophenyl/phosphorylcholine, and production of p-nitrophenol is quantified spectrophotometrically at 410 nm. Protein phosphatase 2A from bovine kidney is incubated in the presence or absence of GW4869 in buffer containing 50 mM Tris, pH 7.4, 1 mM dithiothreitol, 100 μM MnCl2, and 20% glycerol, and phosphatase activity is measured[1]. |
| Cell Research | 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]. |
| Molecular Weight | 577.5 |
| Formula | C30H30Cl2N6O2 |
| Cas No. | 6823-69-4 |
| Smiles | Cl.Cl.O=C(Nc1ccc(cc1)C1=NCCN1)\C=C\c1ccc(\C=C\C(=O)Nc2ccc(cc2)C2=NCCN2)cc1 |
| Relative Density. | no data available |
| Storage | Keep away from moisture, Powder: -20°C for 3 years | In solvent: -80°C for 1 year Shipping with blue ice/Shipping at ambient temperature. | ||||||||||||||||||||
| Solubility Information | DMSO: 5.78 mg/mL (10.01 mM), Sonication is recommended.  | ||||||||||||||||||||
| In Vivo Formulation | PBS: < 1 mg/mL (insoluble) Please add the solvents sequentially, clarifying the solution as much as possible before adding the next one. Dissolve by heating and/or sonication if necessary. Working solution is recommended to be prepared and used immediately. The formulation provided above is for reference purposes only. In vivo formulations may vary and should be modified based on specific experimental conditions. | ||||||||||||||||||||
Solution Preparation Table | |||||||||||||||||||||
DMSO
Note : The dilution table applies only to solid products. For liquid products, please calculate the stock solution based on the stated concentration and/or density. | |||||||||||||||||||||
For example, if the intended dosage is 10 mg/kg for animals weighing 20 g , with a dosing volume of 100 μL per animal, and a total of 10 animals are to be administered, using a formulation of 
10% DMSO+ 40% PEG300+ 5% Tween 80+ 45% Saline/PBS/ddH2O , the resulting working solution concentration would be 2 mg/mL.Dissolve 2 mg of the compound in 100 μL DMSO to obtain a stock solution at a concentration of 20 mg/mL . If the required concentration exceeds the compound's known solubility, please contact us for technical support before proceeding.
1) Add 100 μL of the DMSO stock solution to 400 µL PEG300 and mix thoroughly until the solution becomes clear.
2) Add 50 µL Tween 80 and mix well until fully clarified.
3) Add 450 µL Saline,PBS or ddH2O and mix thoroughly until a homogeneous solution is obtained.
