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Cat. No.: GW4869-1 (for 5mM×0.2ml)

Cat. No.: GW4869-5 (for 5mg)

Cat. No.: GW4869-25 (for 25mg)

Description

Exosome Inhibitor (GW4869) is a cell-permeable, non-competitive inhibitor of neutral sphingomyelinase (N-SMase) that blocks the ceramide-mediated inward budding of multivesicular bodies (MVBs) and the release of mature exosomes from MVBs, thus inhibiting exosome synthesis and release.

Exosomes are membrane-bound extracellular vesicles (EVs) with diameters of approximately 40-160 nm and a lipid bilayer structure. They naturally occur in blood, urine, cerebrospinal fluid, and the supernatant of cultured cells. Almost all cell types can produce and release exosomes. As shown in Figure 1, the cell membrane undergoes endocytosis, sequentially forming early sorting endosomes (ESE), late sorting endosomes (LSE), and MVBs, which contain intraluminal vesicles (ILVs). MVBs fuse with the cell membrane to form exosomes. These exosomes carry various components from their parent cells, including nucleic acids, proteins, lipids, carbohydrates, and metabolites, and release them into the extracellular matrix. Exosomes can be recognized and fused by nearby or distant cells, acting as important mediators of intercellular regulation. They participate in the pathogenesis of various diseases, including cancer, neurodegenerative diseases, and inflammatory diseases, affecting many cellular functions.

Chemical Information

• Chemical Name: 4',4''-Di-2-imidazolin-2-yl-p-benzenediacrylanilide dihydrochloride
• Abbreviation: GW4869
• Aliases: GW69A, GW554869A, GW4869 dihydrochloride, GW4869 2HCl, N-SMase Inhibitor
• Chemical Formula: C30H30Cl2N6O2
• Molecular Weight: 577.50
• CAS Number: 6823-69-4
• Purity: ≥97%
• Solvents/Solubility: DMSO ≥3mg/ml (requires ultrasonic treatment); insoluble in water; insoluble in 0.1M HCl
• Note: GW4869 is usually formulated as a suspension.
• Solution Preparation: Dissolve 5mg in 1.73ml DMSO, using ultrasonic treatment to prepare a 5mM solution.

In Vitro Studies

GW4869 (10 μM) can partially inhibit TNF (Tumor Necrosis Factor)-induced hydrolysis of sphingomyelin, and GW4869 (20 μM) can completely reverse the effect of TNF. GW4869 can dose-dependently reduce TNF-induced mitochondrial cytochrome C release and caspase 9 activation, thereby enhancing cell viability. Exosomes released by bacteria-infected macrophages are pro-inflammatory. GW4869 pretreatment significantly inhibited LPS-induced release of exosomes and pro-inflammatory cytokines (TNF-α, IL-1β, IL-6) from RAW264.7 macrophages. Cancer-associated fibroblasts (CAFs) in pancreatic ductal adenocarcinoma (PDAC) show a high prevalence and certain resistance to Gemcitabine (the standard chemotherapy for PDAC). After treatment with Gemcitabine, the release of exosomes from CAFs significantly increased, whereas the combination of GW4869 and Gemcitabine significantly reduced their survival rate and inhibited their resistance.

In Vivo Studies

In endotoxin-treated or cecal ligation/puncture (CLP)-induced sepsis mouse models, GW4869 pretreatment showed lower serum levels of exosomes and pro-inflammatory cytokines compared to control mice. This significantly alleviated sepsis-induced cardiac inflammation and myocardial depression, extending the survival of mice. In a high-fat diet (HFD)-induced fatty liver mouse model, GW4869 effectively blocked exosome secretion and alleviated the progression of fatty liver disease. In hypertrophic scar formation, M2 macrophages regulate fibroblasts to differentiate into myofibroblasts with active biological functions and proliferation, expressing α-smooth muscle actin (α-SMA). Local injection of GW4869 during the late stage of scar formation significantly inhibited the expression of CD63 and TSG101, reduced the number of α-SMA-positive fibroblasts, and lessened tissue fibrosis after wound healing.

Characteristics

GW4869 acts as a non-competitive inhibitor of N-SMase in vitro but does not inhibit acid sphingomyelinase (A-SMase) even at concentrations up to 150 μM.

Storage

Store at -20ºC:

• Solutions: valid for one year.
• Powders: valid for three years.
• For longer storage, store solutions at -80ºC.

Precautions

• The powder is extremely difficult to dissolve when preparing the solution, requiring prolonged ultrasonic treatment. It is recommended to purchase the pre-made solution.
• This product is intended for scientific research use only by professionals and must not be used for clinical diagnosis or treatment, food or drugs, or stored in a regular household.
• For your safety and health, wear a lab coat and disposable gloves while handling.

Only for research and not intended for treatment of humans or animals

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SBS Genetech is a long-term sponsor of Cold Spring Harbor Laboratory