| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| 10mg |
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| 25mg | |||
| Other Sizes |
| Targets |
The primary targets of 5-Iodo-indirubin-3'-monoxime are the kinases GSK-3beta, CDK5/P25, and CDK1/cyclin B. It acts as an ATP-competitive inhibitor, binding to the ATP-binding pocket of these enzymes. The compound exhibits potent inhibitory activity with IC50 values of 9 nM for GSK-3beta, 20 nM for CDK5/P25, and 25 nM for CDK1/cyclin B. By inhibiting these kinases, it can modulate various signaling pathways, including those involved in cell cycle progression, neuronal survival, and glycogen metabolism.
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| ln Vitro |
In vitro, 5-Iodo-indirubin-3'-monoxime is a potent inhibitor of GSK-3beta, CDK5/P25, and CDK1/cyclin B. Its inhibitory activity is characterized by IC50 values of 9 nM for GSK-3beta, 20 nM for CDK5/P25, and 25 nM for CDK1/cyclin B. It acts as a competitive inhibitor of ATP, binding to the catalytic site of the kinase. This potent and selective inhibition makes it a useful tool for studying the roles of these specific kinases in cell cycle control, neurodegenerative diseases, and other cellular processes. It is often used in biochemical assays to validate kinase targets.
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| ln Vivo |
In vivo, 5-Iodo-indirubin-3'-monoxime is primarily used as a research tool to study the effects of inhibiting GSK-3beta, CDK5, and CDK1 in animal models of disease. Its potent inhibition of CDK1 and CDK5 makes it useful for studying cell cycle regulation and neurodegenerative conditions. By inhibiting GSK-3beta, it can modulate pathways involved in neuroprotection and metabolic regulation. However, specific in vivo efficacy data for this compound are limited, and it is primarily used in preclinical research to probe kinase function and validate therapeutic targets.
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| Enzyme Assay |
Cell-free enzyme assays for 5-Iodo-indirubin-3'-monoxime typically use recombinant human kinases, such as GSK-3beta, CDK5/p25, or CDK1/cyclin B. A standard protocol involves incubating the kinase with a specific peptide substrate, ATP, and varying concentrations of the inhibitor in a kinase buffer. The reaction is carried out at 30degC for 30-60 minutes. The amount of phosphorylated substrate is measured using a luminescent ATP detection assay or by using radiolabeled ATP (e.g., [gamma-33P]-ATP) followed by scintillation counting. The IC50 is determined by plotting the percentage of kinase activity remaining against the inhibitor concentration.
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| Cell Assay |
For in vitro cellular experiments, cell lines of interest (e.g., cancer cells or neurons) are cultured in appropriate media. Cells are treated with 5-Iodo-indirubin-3'-monoxime at various concentrations (typically 0.1-100 uM) for a specified duration (e.g., 24-72 hours). After treatment, cells are harvested, and the effects on kinase activity are assessed. This can be done by Western blot analysis using phospho-specific antibodies against the substrates of the target kinases (e.g., phospho-GSK-3beta, phospho-CDK substrates). Cell viability, proliferation, and apoptosis are also assessed using MTT or flow cytometry.
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| Animal Protocol |
In vivo animal experiments with 5-Iodo-indirubin-3'-monoxime are typically conducted in mouse models of disease. A common protocol involves administering the compound via intraperitoneal (IP) or oral (PO) injection at doses ranging from 1-50 mg/kg. The compound is usually formulated in a vehicle such as DMSO, PEG300, or saline. After treatment, tissues are collected, and target kinase activity is assessed by Western blotting or kinase activity assays. Disease progression is monitored using appropriate endpoints, such as tumor size in cancer models or behavioral tests in neurodegenerative models.
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| ADME/Pharmacokinetics |
5-Iodo-indirubin-3'-monoxime has a molecular weight of 403.17 g/mol and a molecular formula of C16H10IN3O2. It is soluble in DMSO at 66.7 mg/mL (165.43 mM). For in vivo studies, it can be formulated in a vehicle such as 5% DMSO + 30% PEG300 + 5% Tween 80 + 60% ddH2O. The powder form is stable for up to 3 years when stored at -20degC. Solutions should be stored at -80degC for up to 1 year to prevent degradation.
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| Toxicity/Toxicokinetics |
The toxicity profile of 5-Iodo-indirubin-3'-monoxime is not fully characterized in the available literature. As a potent kinase inhibitor, it has the potential for off-target effects and toxicity. However, in vitro studies have shown it to be a potent inhibitor with good selectivity for its targets. In vivo toxicity would need to be assessed in animal models. The compound should be handled with standard laboratory precautions, including the use of personal protective equipment. It is for research use only and not for human therapeutic use.
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| References | |
| Additional Infomation |
5-Iodo-indirubin-3'-monoxime is a potent indirubin derivative that acts as an ATP-competitive inhibitor of GSK-3beta (IC50 = 9 nM), CDK5/P25 (IC50 = 20 nM), and CDK1/cyclin B (IC50 = 25 nM). It is a valuable research tool for studying the roles of these kinases in various biological processes and diseases. Its high potency and defined selectivity make it useful for target validation and mechanistic studies. It is available as a research compound and is not approved for clinical use.
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| Molecular Formula |
C16H10N3O2I
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|---|---|
| Molecular Weight |
403.1735
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| Exact Mass |
402.981
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| CAS # |
331467-03-9
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| PubChem CID |
403176
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| Appearance |
Pink to red solid powder
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| LogP |
3.8
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| Hydrogen Bond Donor Count |
3
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| Hydrogen Bond Acceptor Count |
3
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| Rotatable Bond Count |
1
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| Heavy Atom Count |
22
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| Complexity |
438
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| Defined Atom Stereocenter Count |
0
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| SMILES |
O/N=C1/C2C(=CC=CC=2)N/C/1=C1\C2C(=CC=C(I)C=2)NC\1=O
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| InChi Key |
WCCYEJYTLNWYCC-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C16H10IN3O2/c17-8-5-6-12-10(7-8)13(16(21)19-12)15-14(20-22)9-3-1-2-4-11(9)18-15/h1-7,18-19,21H
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| Chemical Name |
5-iodo-3-(3-nitroso-1H-indol-2-yl)-1H-indol-2-ol
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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 : ~67.5 mg/mL (~167.42 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.25 mg/mL (5.58 mM) (saturation unknown) 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.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 22.5 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. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.4803 mL | 12.4017 mL | 24.8034 mL | |
| 5 mM | 0.4961 mL | 2.4803 mL | 4.9607 mL | |
| 10 mM | 0.2480 mL | 1.2402 mL | 2.4803 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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