GW4869 |
| Catalog No.GC19186 |
GW4869 is a noncompetitive neutral sphingomyelinase(N-SMase) inhibitor with an IC50 of 1 uM.
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 6823-69-4
Sample solution is provided at 25 µL, 10mM.
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 of gw4869 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 the 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 display lower amounts of exosomes and pro-inflammatory cytokines in the serum than control PBS-injected mice. Accordingly, GW4869 treatment diminishes 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 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]. 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]: | |
Cell lines | MCF7 human breast cancer cells |
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. |
Reaction Conditions | 30 min |
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]: | |
Animal models | Male wild-type C57BL/6 mice |
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. |
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. |
References: | |
| Cas No. | 6823-69-4 | SDF | |
| Chemical Name | 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 | ||
| Formula | C30H30Cl2N6O2 | M.Wt | 577.5 |
| Solubility | Water : < 0.1 mg/mL (insoluble); DMSO : 2 mg/mL | Storage | Store at -20°C, ready to use |
| General tips | Please select the appropriate solvent to prepare the stock solution according to the
solubility of the product in different solvents; once the solution is prepared, please store it in
separate packages to avoid product failure caused by repeated freezing and thawing.Storage method
and period of the stock solution: When stored at -80°C, please use it within 6 months; when stored
at -20°C, please use it within 1 month. To increase solubility, heat the tube to 37°C and then oscillate in an ultrasonic bath for some time. |
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| Shipping Condition | Evaluation sample solution: shipped with blue ice. All other sizes available: with RT, or with Blue Ice upon request. | ||
| Prepare stock solution | |||
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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 |
Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)
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Working concentration: mg/ml;
Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid concentration mg/mL, Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug. )
Method for preparing in vivo formulation: Take μL DMSO master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O, mix and clarify.
Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.
Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such as vortex, ultrasound or hot water bath can be used to aid dissolving.
3. All of the above co-solvents are available for purchase on the GlpBio website.
Quality Control & SDS
- View current batch:
- Purity: >98.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
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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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