| Size | Price | |
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| 5mg | ||
| Other Sizes |
| Targets |
Receptor-interacting serine/threonine protein kinase 1 (RIP1, also known as RIPK1). RIP1 is a key regulator of cell death and inflammation, mediating both apoptosis and necroptosis. RIP1 kinase activity is involved in the activation of NF-kappaB and MAPK pathways, contributing to inflammatory responses. In pancreatic cancer, RIP1 kinase drives macrophage-mediated adaptive immune tolerance, allowing tumors to evade immune detection. GSK547 is a selective RIP1 inhibitor that blocks its kinase activity without affecting other RIP family members.
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| ln Vitro |
GSK547 is a potent and selective receptor-interacting serine/threonine protein kinase 1 (RIP1) inhibitor that can suppress macrophage-mediated adaptive immune tolerance in pancreatic cancer. In vitro, GSK547 inhibits RIP1 kinase activity with high potency (IC50 in the low nanomolar range) and shows selectivity over other kinases. Treatment of macrophages with GSK547 blocks RIP1-dependent inflammatory cytokine production and alters macrophage polarization. (Rac)-GSK547 is the racemic form and can be used as a research tool or experimental control.
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| ln Vivo |
In vivo, GSK547 has shown efficacy in the KPC (LSL-KrasG12D/+; LSL-Trp53R172H/+; Pdx-1-Cre) mouse model of pancreatic ductal adenocarcinoma (PDA). In this model, GSK547 treatment suppresses macrophage-mediated adaptive immune tolerance and promotes immunogenic macrophage differentiation, leading to adaptive immune activation and tumor protection. RIP1 inhibition in pancreatic cancer results in reduced tumor growth and enhanced anti-tumor immunity. GSK547 has also been used in the EAE (experimental autoimmune encephalomyelitis) mouse model of multiple sclerosis, where RIP1 inhibition reduces neuroinflammation and disease severity. (Rac)-GSK547 is used as a control compound in these studies.
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| Enzyme Assay |
A RIP1 kinase activity assay is performed in 384-well format using purified recombinant human RIP1 kinase domain. The enzyme is incubated with ATP (1-10 uM) and a fluorescently-labeled peptide substrate or biotinylated substrate in assay buffer (50 mM HEPES, pH 7.4, 10 mM MgCl2, 1 mM DTT, 0.01% Triton X-100) for 30-60 minutes at room temperature. Varying concentrations of GSK547 (0.01 nM to 10 uM) are included to determine IC50 values. The reaction is stopped, and phosphorylation is detected by fluorescence polarization, TR-FRET, or AlphaScreen. IC50 values are calculated from dose-response curves using a four-parameter logistic regression. (Rac)-GSK547 is used as a control.
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| Cell Assay |
For cellular RIP1 activity assays, macrophages (e.g., bone marrow-derived macrophages, BMDMs, or cell lines like THP-1) are seeded in 96-well plates. Cells are primed with TNFalpha (10-50 ng/mL) to activate RIP1-dependent signaling, then treated with varying concentrations of GSK547 or the (Rac)-form as control (0.1 nM to 10 uM) for 1-4 hours. Cell lysates are analyzed by Western blot for phosphorylation of RIP1 substrates (e.g., MLKL) or for NF-kappaB activation markers (p-IkappaB, p-p65). Cytokine production (TNFalpha, IL-6, IL-1beta) is measured by ELISA. Cell viability in necroptosis assays is measured by propidium iodide uptake or LDH release. (Rac)-GSK547 is used as a negative control.
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| Animal Protocol |
For in vivo efficacy in pancreatic cancer, the KPC (LSL-KrasG12D/+; LSL-Trp53R172H/+; Pdx-1-Cre) genetically engineered mouse model is used. KPC mice develop spontaneous pancreatic tumors that recapitulate human PDA. GSK547 is administered orally at doses of 10-100 mg/kg once or twice daily for 2-4 weeks. Tumor growth is monitored by palpation, ultrasound imaging, or at endpoint by tumor weight measurement. Tumor-infiltrating immune cells are analyzed by flow cytometry (macrophage polarization markers M1: CD86, iNOS; M2: CD206, Arg1). For the EAE model, C57BL/6 mice are immunized with MOG35-55 peptide and pertussis toxin to induce EAE. GSK547 (25-100 mg/kg) is administered orally once daily starting at disease onset. Clinical scores (paralysis severity) are assessed daily, and spinal cords are collected for histopathological analysis of demyelination and immune cell infiltration. (Rac)-GSK547 is used as a control compound.
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| ADME/Pharmacokinetics |
(Rac)-GSK547 is the racemate of GSK547 and can be used as a research tool or experimental control. GSK547 is soluble in DMSO at ≥125 mg/mL (315.35 mM). For in vivo studies, GSK547 can be formulated in a vehicle such as 10% DMSO in saline, PEG400/saline, or a mixture of DMSO/PEG300/Tween-80/saline. GSK547 has a LogP of 1.9 and tPSA of 85, indicating favorable drug-like properties. The compound is stable when stored at -20degC under nitrogen; in solvent, it can be stored at -80degC for 6 months or at -20degC for 1 month (stored under nitrogen).
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| Toxicity/Toxicokinetics |
In preclinical studies, GSK547 is well-tolerated at therapeutic doses (10-100 mg/kg) in mice, with no significant body weight loss or overt signs of toxicity. RIP1 inhibitors as a class have been shown to have manageable safety profiles in animal studies. Since RIP1 kinase is involved in inflammatory signaling, inhibition may affect normal immune function, but no significant immunosuppression or increased susceptibility to infections has been reported in short-term studies. Comprehensive toxicology data for GSK547 is not publicly available. (Rac)-GSK547 is for research use only.
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| References | |
| Additional Infomation |
GSK547 was developed by GlaxoSmithKline (GSK) as a potent and selective RIP1 kinase inhibitor for cancer immunotherapy. The discovery and characterization of GSK547 were published in Cancer Cell in 2018, where it was shown that RIP1 kinase drives macrophage-mediated adaptive immune tolerance in pancreatic cancer and that RIP1 inhibition with GSK547 promotes anti-tumor immunity and suppresses tumor growth. GSK547 has become a valuable research tool for studying RIP1-dependent cell death, inflammation, and immune modulation in oncology and autoimmune diseases. This product is for research use only and is not FDA-approved for human therapy.
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| Molecular Formula |
C20H18F2N6O
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|---|---|
| Molecular Weight |
396.39
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| Related CAS # |
GSK547;2226735-55-1
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| Appearance |
White to off-white solid powder
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture. |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
DMSO :≥ 125 mg/mL (~315.35 mM)
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| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.5228 mL | 12.6138 mL | 25.2277 mL | |
| 5 mM | 0.5046 mL | 2.5228 mL | 5.0455 mL | |
| 10 mM | 0.2523 mL | 1.2614 mL | 2.5228 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution 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 stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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