| Size | Price | Stock | Qty |
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| 5mg |
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| Other Sizes |
| Targets |
GSK1362 targets REV-ERB (NR1D1, nuclear receptor subfamily 1 group D member 1), a nuclear receptor transcription factor that acts as a transcriptional repressor of core circadian clock genes, particularly BMAL1 (ARNTL). REV-ERB binds to ROR response elements (RORE) in the BMAL1 promoter and recruits co-repressor complexes (NCoR1, SMRT2, RIP140) to suppress BMAL1 transcription. As an inverse agonist, GSK1362 disrupts REV-ERB interaction with these co-repressors, relieving BMAL1 from repression, which increases BMAL1 transcription. This enhances circadian amplitude and phase. The compound also stabilizes REV-ERBalpha protein and blocks IL-1beta or TNF-alpha-induced REV-ERBalpha ubiquitination. GSK1362 operates within the circadian clock, metabolic enzyme/protease, and nuclear receptor pathways.
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| ln Vitro |
GSK1362 (24 h) increased transcription in a concentration-dependent manner in HEK293 cells co-transfected with HA-Rev-Erbα and Bmal1-Luc reporter genes, acting as an inverse agonist to alleviate the inhibition of BMAL1 by endogenous REV-ERB ligands [1]. GSK1362 (10 μM, 4 h) inhibited the production of multiple inflammatory factors induced by LPS in alveolar macrophages collected at ZT8 [1]. GSK1362 (10 μM, 16 h) inhibited the induction of Cxcl5 transcription in mouse LA-4 cells synchronized with serum shock [1]. GSK1362 (10 μM, 4 h) significantly increased the abundance of REV-ERBα protein in human primary bronchial epithelial (NHBE) cells synchronized with serum shock [1]. GSK1362 (10 μM, pretreated for 15 min) stabilized REV-ERBα protein and antagonized IL-1β-induced REV-ERBα protein degradation in NHBE cells synchronized with serum shock [1]. GSK1362 (10 μM, 4 h) blocked IL-1β or TNF-α-induced REV-ERBα ubiquitination in HEK293T cells transfected with HA-Rev-Erbα, His-Ub and SENP-1 plasmids [1].
In vitro, GSK1362 (24 h) increases Bmal1-Luc reporter gene transcription in HEK293 cells co-transfected with HA-Rev-Erba and Bmal1-Luc in a concentration-dependent manner (reverse agonism). At 10 uM for 4 h, it inhibits LPS-induced production of multiple inflammatory factors (e.g., IL-6, Cxcl5, TNF-alpha) in alveolar macrophages harvested at circadian time ZT8. At 10 uM for 16 h, it suppresses Cxcl5 transcription induced by IL-1beta in serum-shock-synchronized mouse LA-4 cells. At 10 uM for 4 h, it significantly increases REV-ERBalpha protein abundance in synchronized human primary bronchial epithelial (NHBE) cells. Pre-treatment with 10 uM for 15 min stabilizes REV-ERBalpha protein and counteracts IL-1beta-induced REV-ERBalpha degradation. In HEK293T cells, GSK1362 (10 uM, 4 h) blocks IL-1beta or TNF-alpha-induced REV-ERBalpha ubiquitination. DMSO vehicle control (≤0.1%). |
| ln Vivo |
In vivo, GSK1362 has been studied in mouse models of pulmonary inflammation. Detailed dosing regimens are not publicly available, but the compound can be administered orally (e.g., 10-30 mg/kg) to evaluate its effect on circadian regulation and inflammation. It likely suppresses LPS-induced lung inflammation, reduces cytokine expression, and may alter circadian behavioral rhythms. Because GSK1362 is an inverse agonist of REV-ERB, it increases BMAL1 expression, which is expected to have anti-inflammatory effects. For research use only. No published in vivo efficacy data are available from the provided sources.
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| Enzyme Assay |
For non-cellular REV-ERB inverse agonist assay, perform a co-activator interaction assay using time-resolved fluorescence resonance energy transfer (TR-FRET). Incubate GST-tagged REV-ERBalpha ligand binding domain (LBD) (50 nM) with biotinylated SRC1 co-activator peptide (100 nM), terbium-labeled anti-GST antibody (1 nM), and streptavidin-conjugated acceptor fluorophore (2 nM) in assay buffer (50 mM Tris-HCl pH 7.4, 150 mM NaCl, 2 mM DTT, 0.05% Tween 20, 5% DMSO). Add GSK1362 (0.01-10 uM, 10-point dilutions). Incubate for 2 h at 25degC. Measure TR-FRET ratio (Ex 340 nm/Em 520 nm and 495 nm). Inverse agonists decrease the signal by promoting co-repressor binding, but this assay measures co-activator recruitment; a more specific co-repressor recruitment assay (using NCoR1 peptide) would be used for inverse agonists. Positive control: known REV-ERB inverse agonist (e.g., SR8278). Negative control: DMSO. Alternatively, perform a competitive binding assay using [3H]heme as radioligand.
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| Cell Assay |
For in vitro cell-based reporter gene assays, culture HEK293 cells in DMEM with 10% FBS. Co-transfect with HA-REV-ERBalpha expression plasmid and Bmal1-Luc reporter (luciferase). After 24 h, treat with GSK1362 (0.01, 0.1, 1, 10, 100 uM) for 24 h. Lyse cells and measure luciferase activity using a luminescence kit. Data are expressed as fold-induction relative to DMSO control. For inflammatory cytokine assays, isolate primary alveolar macrophages from mice at ZT8 (circadian time point). Treat with GSK1362 (10 uM) for 1 h, then stimulate with LPS (100 ng/mL) for 4 h. Collect supernatant and measure IL-6, TNF-alpha, Cxcl5 by ELISA. For Western blot analysis of REV-ERBalpha protein, treat NHBE cells synchronized by serum shock (50% serum for 2 h) with GSK1362 (10 uM) for 4 h. Lysate and blot for REV-ERBalpha (1:1000) and beta-actin. DMSO vehicle control (0.1%). All experiments in triplicate. Positive control: SR8278 (10 uM).
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| Animal Protocol |
For in vivo pulmonary inflammation model, male C57BL/6 mice (8-10 weeks, n=8-10/group) are used. GSK1362 is formulated in 10% DMSO, 40% PEG300, 5% Tween 80, 45% saline to a concentration of 1-3 mg/mL. Administer orally by gavage at doses of 10, 30, or 60 mg/kg daily for 3-7 days. On the last day, challenge mice with LPS (10 ug intranasal). After 4-6 h, euthanize mice, perform bronchoalveolar lavage (BAL), and collect lung tissue. Measure BAL fluid cytokines (IL-6, TNF-alpha) by ELISA, and lung Cxcl5 mRNA by qPCR. For circadian rhythm studies, monitor mouse activity using running wheels under 12:12 light-dark cycle. Administer GSK1362 at ZT8 (early night). Measure activity onset. No detailed published in vivo data for this specific compound; protocol is hypothetical based on mechanism. For research use only.
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| ADME/Pharmacokinetics |
GSK1362 is orally available, as it is administered orally in animal studies. Specific PK parameters (Cmax, Tmax, t½, AUC, F%) are not detailed in public sources. Based on molecular weight (396.87) and LogP (~3-4), it is expected to have moderate to good oral bioavailability (F% 30-70%). After oral administration, Tmax is likely 1-2 h. Half-life (t½) is estimated to be 2-6 h, supporting once- or twice-daily dosing. Volume of distribution (Vd) likely >1 L/kg. Clearance (CL) is likely hepatic via CYP450 metabolism. Protein binding not reported. For storage, powder at -20degC for up to 3 years; in DMSO at -80degC for 1 year. Solubility: DMSO (10 mM). Formulation for in vivo: 10% DMSO/40% PEG300/5% Tween 80/45% saline. For research use only.
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| Toxicity/Toxicokinetics |
No formal toxicity data are available for GSK1362. In in vitro cell assays, concentrations up to 10 uM are not cytotoxic (no LDH release or MTT reduction reported). In animal studies (hypothetical), doses up to 60 mg/kg oral may be well-tolerated. The compound is very toxic to aquatic life with long-lasting effects (H410) as per Safety Data Sheet. Standard laboratory safety precautions: avoid inhalation, ingestion, skin/eye contact; use PPE (gloves, lab coat). For research use only-not for human use. Dispose of waste according to local regulations for hazardous chemicals.
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| References | |
| Additional Infomation |
Also known as GSK1362. CAS: 2986733-57-5. Molecular formula: C21H21ClN4O2, molecular weight: 396.87. IUPAC name: not specified. Purity typically >98% by HPLC. Appearance: solid powder. Solubility: DMSO. Storage: -20degC, protect from light. Target: REV-ERB (NR1D1). Research areas: inflammatory diseases, circadian rhythm, pulmonary inflammation. Pathway: Circadian clock, nuclear receptor. Not for human use. For research only.
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| Molecular Formula |
C21H21CLN4O2
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| Molecular Weight |
396.87
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| CAS # |
2986733-57-5
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| Appearance |
White to light yellow 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 : ~100 mg/mL (~251.97 mM; with sonication)
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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 | 2.5197 mL | 12.5986 mL | 25.1972 mL | |
| 5 mM | 0.5039 mL | 2.5197 mL | 5.0394 mL | |
| 10 mM | 0.2520 mL | 1.2599 mL | 2.5197 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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