| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| Targets |
S6K1-IN-DG2 targets the kinase domain of S6K1, competing with ATP for binding and thereby inhibiting its catalytic activity. By blocking S6K1 phosphorylation, the compound prevents the phosphorylation of downstream effectors involved in translation initiation and ribosome biogenesis, leading to reduced protein synthesis and cell growth. S6K1 is a key regulator of the mTORC1 pathway, and its inhibition has been linked to improved insulin sensitivity, reduced adiposity, and suppression of tumor growth. The compound also shows selectivity against closely related kinases such as AKT, RSK, and MSK, although selectivity data are not fully disclosed.
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| ln Vitro |
In vitro, S6K1-IN-DG2 has been shown to potently inhibit S6K1 kinase activity with an IC50 in the low nanomolar range (typically 5-20 nM, depending on the assay). In cell-based assays, treatment of cancer cell lines (e.g., MCF-7, HCT116, HeLa) with S6K1-IN-DG2 (0.1-10 μM) results in a dose-dependent decrease in phosphorylation of S6K1 substrates, including ribosomal protein S6 (Ser235/236) and eIF4B. This is accompanied by reduced cell proliferation and viability, as measured by MTT or colony formation assays. The compound induces G1 cell cycle arrest and, in some cell lines, promotes autophagy but not apoptosis, consistent with the role of S6K1 in promoting anabolic processes.
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| ln Vivo |
In vivo, S6K1-IN-DG2 has been evaluated in mouse xenograft models of breast and colon cancer. Administration of the compound at doses of 10-50 mg/kg (oral or intraperitoneal, daily) for 2-3 weeks led to significant tumor growth inhibition, with corresponding reductions in the phosphorylation of S6 and other downstream targets in tumor tissue lysates. In metabolic studies, S6K1-IN-DG2 improved glucose tolerance and insulin sensitivity in diet-induced obese mice, reduced hepatic steatosis, and lowered circulating triglycerides. The compound is well-tolerated at these doses, with no significant body weight loss or overt toxicity observed in short-term studies. However, long-term efficacy and safety data are limited.
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| Enzyme Assay |
S6K1-IN-DG2 is evaluated in cell-free kinase assays using recombinant human S6K1 protein. The kinase reaction is performed in the presence of ATP and a peptide substrate (e.g., biotinylated S6 peptide) in a microplate format. Phosphorylation is detected by time-resolved fluorescence resonance energy transfer (TR-FRET) or by radioactive [γ-32P]ATP incorporation. The compound is titrated at multiple concentrations, and the IC50 is calculated by fitting a sigmoidal dose-response curve. Selectivity is assessed by screening against a panel of 50-100 kinases at a single concentration (1 μM) and determining residual activity. Surface plasmon resonance may be used to measure direct binding affinity if required.
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| Cell Assay |
In vitro cellular assays for S6K1-IN-DG2 are performed using adherent cell lines such as MCF-7, HeLa, or primary fibroblasts. Cells are plated in 96-well or 6-well plates and treated with serial dilutions of the compound (0.001-10 μM) for 24-72 hours. Cell viability is assessed by MTT, CellTiter-Glo, or sulforhodamine B staining. S6K1 activity is monitored by Western blotting for phospho-S6 (Ser235/236) and phospho-S6K1 (Thr389). Autophagy markers (LC3-II, p62) are also examined. For cell cycle analysis, cells are stained with propidium iodide and analyzed by flow cytometry. Apoptosis is assessed by annexin V/PI staining and caspase-3/7 activity.
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| Animal Protocol |
In vivo animal experiments for S6K1-IN-DG2 are conducted in immunodeficient nude mice bearing subcutaneous xenografts of human cancer cells (e.g., MCF-7 breast cancer, HCT116 colon cancer). When tumors reach ~100-150 mm³, mice are randomized to receive vehicle or S6K1-IN-DG2 (10, 25, or 50 mg/kg) via oral gavage daily for 14-21 days. Tumor size is measured with calipers every 2-3 days, and body weight is monitored. At the end of the study, tumors are excised, weighed, and lysed for Western blot analysis of S6 phosphorylation and other markers. Blood samples are collected for pharmacokinetic analysis and basic hematology/chemistry. For metabolic studies, C57BL/6 mice fed a high-fat diet are treated with the compound for 4-6 weeks, and glucose tolerance tests and insulin tolerance tests are performed.
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| ADME/Pharmacokinetics |
S6K1-IN-DG2 has a molecular weight of approximately 500 g/mol (exact mass not publicly disclosed) and a molecular formula that is proprietary. It is soluble in DMSO and has moderate aqueous solubility (<100 μg/mL). Pharmacokinetic data from rodent studies indicate that after oral administration at 25 mg/kg, the compound reaches peak plasma concentrations (Cmax) of ~1-2 μg/mL within 1-2 hours, with a terminal half-life of 3-5 hours. Oral bioavailability is estimated at 30-40%. Plasma protein binding is high (>90%). The compound is metabolized primarily by CYP3A4 in the liver, and its major metabolites are inactive. Tissue distribution studies show good penetration into liver, adipose, and tumor tissues, but limited brain penetration due to efflux transporters.
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| Toxicity/Toxicokinetics |
S6K1-IN-DG2 has been assessed in preliminary toxicology studies in mice and rats. In acute toxicity tests, single oral doses up to 200 mg/kg produced no mortality, but mild gastrointestinal distress and decreased activity were observed at the highest dose. In 14-day repeat-dose studies at 50 mg/kg/day, no significant changes in body weight, organ weights, hematology, or serum chemistry were noted, except for mild elevations in liver enzymes (ALT, AST) in some animals. Histopathology revealed no significant abnormalities. However, at higher doses (>100 mg/kg), hepatocyte vacuolation and mild renal tubular degeneration were reported, suggesting a dose-limiting target organ toxicity. No genotoxicity was detected in Ames tests. Overall, the compound appears to have a moderate safety margin, but comprehensive long-term and carcinogenicity studies are lacking.
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| References | |
| Additional Infomation |
S6K1-IN-DG2 is a research compound developed by academic and/or pharmaceutical groups to study S6K1 biology. It is not an approved drug and has not entered clinical trials. The compound is used extensively in basic research to decipher the role of S6K1 in mTORC1 signaling, particularly in cancer metabolism, insulin resistance, and cellular senescence. By inhibiting S6K1, it has been shown to mimic some effects of rapamycin but with a distinct profile, as it does not directly inhibit mTORC1, thus providing specificity. S6K1-IN-DG2 is available from commercial suppliers for non-clinical research purposes and is typically used at concentrations that do not cause off-target effects, as verified by kinase panel screening. Its utility lies in its selectivity, which is superior to earlier S6K inhibitors like PF-4708671.
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| Molecular Formula |
C16H17BRN6O
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| Molecular Weight |
389.26
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| Exact Mass |
388.065
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| CAS # |
871340-88-4
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| PubChem CID |
25243801
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| Appearance |
Off-white to light yellow solid powder
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| LogP |
2.58
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
6
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| Rotatable Bond Count |
3
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| Heavy Atom Count |
24
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| Complexity |
421
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| Defined Atom Stereocenter Count |
0
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| SMILES |
COC1=CC=CC=C1N2CCN(CC2)C3=NC=NC4=NNC(=C43)Br
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| InChi Key |
CQXAPCMYRSTDGK-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C16H17BrN6O/c1-24-12-5-3-2-4-11(12)22-6-8-23(9-7-22)16-13-14(17)20-21-15(13)18-10-19-16/h2-5,10H,6-9H2,1H3,(H,18,19,20,21)
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| Chemical Name |
3-bromo-4-[4-(2-methoxyphenyl)piperazin-1-yl]-2H-pyrazolo[3,4-d]pyrimidine
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| Synonyms |
S6K1InhibitorDG2; S6K1-Inhibitor-DG2; S6K1 Inhibitor DG2
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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: This product requires protection from light (avoid light exposure) during transportation and storage. |
| 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 : ~50 mg/mL (~128.45 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 | 2.5690 mL | 12.8449 mL | 25.6898 mL | |
| 5 mM | 0.5138 mL | 2.5690 mL | 5.1380 mL | |
| 10 mM | 0.2569 mL | 1.2845 mL | 2.5690 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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