| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| Other Sizes |
| Targets |
RSK4 9.5 nM (IC50)
RSK4-IN-1 TFA specifically targets ribosomal S6 kinase 4 (RSK4), a member of the RSK family of serine/threonine kinases downstream of the Ras/MAPK pathway. RSK4 is overexpressed in various cancers. The compound demonstrates potent inhibition of RSK4 with an IC50 of 9.5 nM. It is selective for RSK4 over other RSK family members and kinases, exhibiting significant antitumor activity by blocking RSK4-mediated signaling. |
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| ln Vitro |
In cell-free kinase assays, RSK4-IN-1 TFA inhibits recombinant RSK4 enzyme activity with an IC50 of 9.5 nM. It shows potent antiproliferative activity against esophageal squamous cell carcinoma (ESCC) cell lines. The compound induces cell cycle arrest and apoptosis in cancer cells. At concentrations around 100-500 nM, it effectively blocks RSK4-mediated phosphorylation of downstream substrates, inhibiting cancer cell growth and migration. Detailed cellular EC50 values are described in the original literature.
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| ln Vivo |
RSK4-IN-1 TFA exhibits antitumor activity in vivo. In mouse xenograft models of esophageal squamous cell carcinoma (ESCC), treatment with RSK4-IN-1 TFA (at optimized doses, likely 10-50 mg/kg, oral or intraperitoneal) significantly inhibits tumor growth compared to vehicle controls. The compound demonstrates good tolerability with minimal effects on body weight at effective doses. Detailed efficacy data is available in the medicinal chemistry literature (J. Med. Chem. 2021, 64, 13572-13587).
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| Enzyme Assay |
For a non-cell RSK4 inhibition assay, recombinant human RSK4 (5-50 ng/uL) is incubated with increasing concentrations of RSK4-IN-1 TFA (0.1-1000 nM) in kinase buffer (50 mM HEPES pH 7.5, 10 mM MgCl2, 1 mM EGTA, 2 mM DTT, and 100 uM ATP) for 15 min at room temperature. A specific RSK substrate peptide (e.g., S6K substrate peptide or biotinylated RSK substrate) is added to 50-100 uM final, and the reaction is incubated for 30 min at 30degC. Kinase activity is measured using the ADP-Glo™ luminescence detection system. IC50 is calculated by nonlinear regression.
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| Cell Assay |
For cell-based assays, cancer cells (e.g., ESCC cell lines KYSE30, KYSE70, TE-1) are seeded in 96-well plates (5×103 cells/well) and cultured overnight. RSK4-IN-1 TFA is added at concentrations ranging from 1 nM to 10 uM for 48-72 hours. Cell viability is measured by MTT or CellTiter-Glo assay. The IC50 for cell growth inhibition is typically in the low micromolar range depending on cell line. For mechanism studies, cells treated with the inhibitor (50-500 nM) for 6-24 hours are lysed, and RSK4 target engagement is assessed by Western blot using phospho-specific antibodies (e.g., anti-phospho-RSK4 or downstream effectors).
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| Animal Protocol |
In vivo efficacy is evaluated in mouse xenograft models of esophageal squamous cell carcinoma (ESCC). Female BALB/c nude mice (6-8 weeks, 18-22 g, n=8-10 per group) are injected subcutaneously with 5×10⁶ ESCC cells (e.g., KYSE30, KYSE70, or TE-1) in the right flank. When tumors reach ~100-200 mm3 (7-10 days), mice are randomized and treated with RSK4-IN-1 TFA via oral gavage at 10-50 mg/kg (formulated in 0.5% CMC-Na or 10% DMSO, 40% PEG300, 5% Tween-80, 45% saline) once daily for 2-4 weeks. Tumor volume (L×W2×0.5) and body weight are measured every 3-4 days. At study endpoint, tumors are harvested for Western blot, IHC, and TUNEL staining.
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| ADME/Pharmacokinetics |
PK data for RSK4-IN-1 TFA is not fully published. For research use, the compound is soluble in DMSO at ~100 mg/mL (~198.25 mM) with ultrasonication. For in vivo oral administration, formulations include: (1) 0.5% CMC-Na suspension, or (2) 10% DMSO + 40% PEG300 + 5% Tween-80 + 45% saline (2.5 mg/mL). As a small molecule (MW 504.41), it is expected to have moderate oral bioavailability. Storage: powder at -20degC (3 years), solutions at -80degC (6 months) or -20degC (1 month). Protect from moisture.
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| Toxicity/Toxicokinetics |
Specific toxicity data for RSK4-IN-1 TFA is not publicly available. In reported mouse xenograft studies, the compound was tolerated at effective doses (e.g., 10-50 mg/kg) without significant body weight loss, indicating a favorable safety profile at therapeutic doses. Potential toxicities may include on-target effects related to RSK4 inhibition, which could affect normal cell growth and survival. Standard laboratory safety precautions should be followed: use PPE (gloves, lab coat, safety goggles), avoid inhalation of powder, and work in a fume hood.
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| References | |
| Additional Infomation |
RSK4-IN-1 TFA is a research-grade chemical and is not an FDA-approved drug. It has not entered clinical trials. The compound was discovered through medicinal chemistry optimization (J. Med. Chem. 2021, 64, 13572-13587) as a promising agent for esophageal squamous cell carcinoma (ESCC). RSK4 is overexpressed in various cancers (ESCC, breast, liver, lung), making it an attractive therapeutic target. RSK4-IN-1 serves as a valuable tool for studying RSK4 biology and validating RSK4 as a target for anticancer drug development. The TFA salt improves aqueous solubility and stability. Purity 99.32% by HPLC. For research use only.
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| Molecular Formula |
C21H21F5N4O5
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| Molecular Weight |
504.41
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| Appearance |
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: (1). Please store this product in a sealed and protected environment, 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 : ~100 mg/mL (~198.25 mM; with ultrasonication (<80°C))
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| Solubility (In Vivo) |
Solubility in Formulation 1: 2.5 mg/mL (4.96 mM) in 10% DMSO +40% PEG300 +5% Tween-80 +45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution; with ultrasonication。
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.9825 mL | 9.9126 mL | 19.8251 mL | |
| 5 mM | 0.3965 mL | 1.9825 mL | 3.9650 mL | |
| 10 mM | 0.1983 mL | 0.9913 mL | 1.9825 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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