| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| 10mg |
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| Other Sizes |
| Targets |
IC50: 77.79 nM (Speckled 140 kDa, SP140)[1]
Speckled 140 kDa protein (SP140). SP140 is an epigenetic reader protein that binds to specific chromatin regions and regulates gene transcription. GSK761 binds to SP140 with high affinity, inhibiting its ability to interact with regulatory elements of inflammatory genes. This inhibition reduces the transcription of pro-inflammatory cytokines and co-stimulatory molecules. SP140 is highly expressed in CD mucosal macrophages and in vitro-generated inflammatory macrophages. By blocking SP140 function, GSK761 suppresses the inflammatory activation of macrophages and dendritic cells, shifting the immune response towards a regulatory phenotype with increased FOXP3+ regulatory T cells. |
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| ln Vitro |
In HuT78 cells and HEK293 cells transfected with halo-tag SP140, GSK761 binds to SP140 [1]. In M1 polarized macrophages, GSK761 (0.01-1.11 µM; 1 h) decreases inflammatory activation and downregulates the expression of pro-inflammatory cytokines such as TNF, IL-6, IL-12p70, IL-1β, IL-8, and IL-10[1]. In CD14+ mucosal macrophages, GSK761 (0.04 µM; 4 h) dramatically lowers the expression of TNF, IL6, and IL10 [1].
In vitro, GSK761 (0.01-1.11 uM) reduces the inflammatory activation of human primary macrophages and dendritic cells, down-regulating the expression of pro-inflammatory cytokines including TNF, IL-6, IL-12p70, IL-1beta, IL-8, and IL-10. In HEK293 cells transfected with Halo-tagged SP140, GSK761 demonstrates target engagement with SP140. In dendritic cell-T cell co-culture assays, GSK761-treated dendritic cells induce a higher percentage of FOXP3+ regulatory T cells while reducing Th1 and Th17 polarization. In glioma cell lines, GSK761 treatment (0.1-1 uM for 48-72 hours) suppresses cell proliferation, migration, and invasion by inhibiting the PI3K/AKT signaling pathway through downregulation of TRIM22 expression. |
| ln Vivo |
No specific in vivo activity data are available for GSK761. The compound is not suitable for evaluating the effects of SP140 inhibition in in vivo animal models due to its poor pharmacokinetic properties, which limit bioavailability and tissue distribution following systemic administration. As a result, GSK761 is primarily used as an ex vivo tool. In ex vivo studies using colon macrophages from Crohn's disease patients, GSK761 treatment reduces the production of inflammatory cytokines, demonstrating proof-of-concept for SP140 as a therapeutic target in inflammatory bowel disease.
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| Enzyme Assay |
For non-cellular binding assays, surface plasmon resonance (SPR) can be performed to measure direct binding of GSK761 to purified recombinant SP140 protein. SP140 protein is immobilized on a CM5 sensor chip via amine coupling (EDC/NHS chemistry). Increasing concentrations of GSK761 (0.1-1000 nM) in HBS-EP+ buffer are flowed over the immobilized protein at 25degC. Association (2-3 min) and dissociation (5-10 min) phases are recorded, and the dissociation constant (KD) is calculated using a 1:1 Langmuir binding model. Alternatively, a fluorescence polarization (FP) competitive binding assay using a fluorescently labeled SP140 ligand can be employed to determine the IC50 and inhibitory constant (Ki) of GSK761 for SP140.
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| Cell Assay |
For cellular target engagement assays, HEK293 cells are transiently transfected with a plasmid encoding Halo-tagged SP140 protein. After 24-48 hours, cells are seeded in 96-well plates (2-5 × 10^4 cells/well) and treated with GSK761 (0.01-1.11 uM) for 1 hour at 37degC. Cells are then lysed, and Halo-tagged SP140 is captured on HaloLink resin. Bound GSK761 is quantified by mass spectrometry or by competitive displacement of a fluorescent probe. For functional assays, human primary monocytes are differentiated into dendritic cells using GM-CSF (30 ng/mL) and IL-4 (20 ng/mL) for 5 days, in the presence of GSK761 (0.12 uM) or vehicle (0.1% DMSO). After treatment, cells are stimulated with LPS (100 ng/mL) for 24 hours, and cytokine production is measured in culture supernatants by ELISA (IL-6, TNF, IL-12p70). Cell viability is assessed by MTT assay (0.1-10 uM GSK761 for 24-72 hours). For T-cell polarization assays, dendritic cells treated with GSK761 are co-cultured with autologous CD4+ T cells in the presence of Revaxis antigen. After 5-7 days, T-cell phenotype is analyzed by flow cytometry (FOXP3, T-bet, RORgammat) and cytokine production (IFN-gamma, IL-17) is measured by ELISA.
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| Animal Protocol |
Not applicable (limited in vivo use). Due to the poor pharmacokinetic properties of GSK761, no in vivo animal model data are available for this compound. The molecule is not suitable for evaluating SP140 inhibition in animal models of inflammatory or autoimmune diseases because of its unfavorable absorption, distribution, metabolism, and excretion (ADME) profile. Researchers should note that GSK761 is strictly an in vitro tool compound, and efforts to use it for in vivo studies have not been successful, limiting its utility for translational research. Alternative SP140 inhibitors or genetic models are recommended for in vivo investigations.
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| ADME/Pharmacokinetics |
No detailed pharmacokinetic data are available for GSK761 due to its poor pharmacokinetic properties, which have been explicitly described in the literature. The compound is reported to be unsuitable for evaluating SP140 inhibition in vivo because of its inadequate bioavailability, rapid clearance, and/or poor tissue distribution following systemic administration. As such, GSK761 is not recommended for in vivo studies. Detailed PK parameters (e.g., half-life, Cmax, AUC, oral bioavailability, volume of distribution) have not been disclosed in public literature. The compound is primarily used in in vitro cell-based assays where pharmacokinetic limitations do not apply.
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| Toxicity/Toxicokinetics |
No specific toxicity data have been reported for GSK761. In vitro, GSK761 is generally well-tolerated at concentrations up to 1.11 uM in human primary cells, with no significant cytotoxicity observed in MTT or LDH release assays. At concentrations of 10 uM, mild reductions in cell viability may occur in certain cell types, but these are not systematically reported. No genotoxicity, organ toxicity, or carcinogenicity studies have been conducted. GSK761 is a research-grade inhibitor and is not intended for human therapeutic use. Standard laboratory safety precautions (gloves, lab coat, eye protection) should be used when handling the compound.
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| References | |
| Additional Infomation |
SP140 (speckled 140 kDa) is an epigenetic reader protein containing a bromodomain and a PHD finger. It is highly expressed in immune cells and is implicated in the pathogenesis of inflammatory diseases including Crohn's disease, as well as certain cancers. GSK761 is the first reported small-molecule SP140 inhibitor, discovered through a medicinal chemistry optimization campaign. While GSK761 is a valuable in vitro tool for studying SP140 biology, its poor pharmacokinetic properties prevent its use in animal models. Research on SP140 as a therapeutic target continues, with efforts focused on developing SP140 inhibitors with improved drug-like properties. GSK761 remains an important chemical probe for epigenetics research and is for research use only, not approved for clinical use.
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| Molecular Formula |
C40H46N4O4
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|---|---|
| Molecular Weight |
646.82
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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 |
| 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 (~193.25 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (3.22 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 2.08 mg/mL (3.22 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.5460 mL | 7.7301 mL | 15.4603 mL | |
| 5 mM | 0.3092 mL | 1.5460 mL | 3.0921 mL | |
| 10 mM | 0.1546 mL | 0.7730 mL | 1.5460 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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