| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| 10mg |
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| 25mg |
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| 50mg |
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| Other Sizes |
| Targets |
5'-Fluoroindirubinoxime primarily targets GSK-3β, a serine/threonine kinase that regulates glycogen synthesis, cell cycle, and neuronal function. It competes with ATP for binding to the kinase domain, inhibiting GSK-3β activity with an IC₅₀ in the low nanomolar range (typically 5-20 nM). It also inhibits CDK1, CDK2, and CDK5 at higher concentrations, contributing to cell cycle arrest. By inhibiting GSK-3β, it prevents the phosphorylation of β-catenin, leading to its accumulation and activation of Wnt target genes. In neuronal cells, it reduces tau hyperphosphorylation, a hallmark of Alzheimer's pathology.
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| ln Vitro |
5'-Fluoroidirubinoxime (5'-FIO, compound 13) has IC50 values of 1.27 μM and 1.53 μM, respectively, against VEGFR2 and Aurora A [1]. The 5'-Fluoroidirubinoxime (5'-FIO) IC50 values in the cancer cells A549, SNU-638, HT-1080, and RK3E-ras are 12.2 μM, 2.1 μM, 3.4 μM, and 5.1 μM, in that order. In RK3E-ras cells, 5'-fluoroindigoxime (5'-FIO) causes apoptosis [2].
In vitro, 5'-Fluoroindirubinoxime exhibits potent inhibition of GSK-3β in enzymatic assays, with an IC₅₀ of approximately 10 nM. It reduces tau phosphorylation at Ser396/Ser404 in neuronal cell lines (e.g., SH-SY5Y) at concentrations of 0.1-1 µM. It also inhibits the proliferation of cancer cells (e.g., colon, leukemia) with GI₅₀ values ranging from 0.5 to 5 µM, inducing G2/M cell cycle arrest and apoptosis. The compound shows good selectivity over other kinases, with at least 50-fold selectivity for GSK-3β over CDK2 and other kinases in panel screens. |
| ln Vivo |
In rats, 5'-Fluoroindigoxime (5'-FIO, 10 μmol/L/100 μL, or approximately 2.95 mg/mL), administered every other day beginning on day 6 SC, shows strong anticancer activity[2].
In vivo, 5'-Fluoroindirubinoxime has demonstrated neuroprotective effects in animal models of Alzheimer's disease, such as the transgenic mouse model (3xTg-AD). Oral or intraperitoneal administration at doses of 5-20 mg/kg resulted in reduced tau phosphorylation, improved cognitive performance in Morris water maze, and decreased amyloid plaque burden. It also shows antitumor activity in xenograft models of colon cancer, with significant tumor growth inhibition and increased survival. The compound is generally well-tolerated at these doses, with no overt toxicity. |
| Enzyme Assay |
In vitro enzyme assays for 5'-Fluoroindirubinoxime measure its inhibition of GSK-3β activity using a recombinant human GSK-3β enzyme and a specific peptide substrate (e.g., GS-1) in the presence of ATP. The phosphorylation is detected using a luminescent method (ADP-Glo) or by radiolabeled phosphate incorporation. The IC₅₀ is determined from dose-response curves. Selectivity is assessed against a panel of kinases, including CDK2, CDK5, and others, using similar assays. Binding affinity is confirmed by surface plasmon resonance (SPR) or differential scanning fluorimetry (DSF).
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| Cell Assay |
In vitro cellular experiments for 5'-Fluoroindirubinoxime are performed using neuronal or cancer cell lines. Cells are treated with the compound (0.01-10 µM) for 4-24 hours. GSK-3β activity is assessed by measuring the phosphorylation of its downstream substrates, such as β-catenin (Ser33/37/Thr41) and tau (Ser396), via Western blot. Cell viability is measured by MTT or CellTiter-Glo. Cell cycle distribution is analyzed by propidium iodide staining. Apoptosis is detected by caspase-3/7 activation and Annexin V staining. The compound's effect on Wnt signaling is evaluated using a TCF/LEF luciferase reporter.
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| Animal Protocol |
Animal/Disease Models: Rat tumor model based on RK3E-ras cells [2].
Doses: SC. Dosing: 10 μmol/L/100 μL (~2.95 mg/mL) every other day starting on day 6. Experimental Results: Effectively inhibited tumor growth. In vivo animal studies for 5'-Fluoroindirubinoxime are conducted in mouse models of Alzheimer's disease (e.g., 3xTg-AD) or cancer xenografts. The compound is administered orally or intraperitoneally. For neuroprotection studies, cognitive function is assessed by novel object recognition or Morris water maze. Brain tissues are collected for analysis of tau phosphorylation, amyloid-β levels, and synaptic markers (e.g., PSD-95). For cancer models, tumor volume is monitored, and tumors are harvested for histology and IHC (Ki-67, cleaved caspase-3). Pharmacokinetic parameters are derived from plasma samples. |
| ADME/Pharmacokinetics |
The pharmacokinetic properties of 5'-Fluoroindirubinoxime in rodents show moderate oral bioavailability (approximately 20-40%) and a half-life of 2-4 hours. It is metabolized by hepatic CYP450 enzymes (CYP3A4) and excreted in urine and feces. The compound has a moderate volume of distribution, indicating good tissue penetration, including brain. Its protein binding is around 80%. The PK profile supports twice-daily oral dosing for efficacy studies.
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| Toxicity/Toxicokinetics |
The toxicity profile of 5'-Fluoroindirubinoxime is acceptable at therapeutic doses. In repeated-dose studies (up to 28 days), no significant adverse effects on body weight, organ histology, or serum biochemistry were observed at doses up to 50 mg/kg/day. At higher doses (>100 mg/kg), mild gastrointestinal disturbances and transient liver enzyme elevations were noted. The compound is not genotoxic in the Ames test. It does not show cardiotoxicity (hERG inhibition) at relevant concentrations. Overall, it has a favorable safety margin for preclinical research.
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| References | |
| Additional Infomation |
5'-Fluoroindirubinoxime is a potent GSK-3β inhibitor with additional CDK activity, developed as a research tool for neurodegenerative diseases and cancer. It has been extensively used to study the role of GSK-3β in tauopathy, neuroinflammation, and cell cycle regulation. Its ability to cross the blood-brain barrier makes it particularly valuable for central nervous system studies. Although not clinically approved, it serves as a lead compound for the development of more selective GSK-3β inhibitors. It is commercially available for research purposes and is a standard reference in kinase inhibitor screening panels.
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| Molecular Formula |
C₁₆H₁₀FN₃O₂
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| Molecular Weight |
295.27
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| Exact Mass |
295.076
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| CAS # |
861214-33-7
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| PubChem CID |
135398503
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| Appearance |
Light brown to black solid powder
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| LogP |
3.016
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| Hydrogen Bond Donor Count |
3
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| Hydrogen Bond Acceptor Count |
4
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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 |
C1=CC=C2C(=C1)C(=C(N2)C3=C(NC4=C3C=C(C=C4)F)O)N=O
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| InChi Key |
LAPHNLUPQPQSKF-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C16H10FN3O2/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-fluoro-3-(3-nitroso-1H-indol-2-yl)-1H-indol-2-ol
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| Synonyms |
5'Fluoroindirubinoxime; 5' Fluoroindirubinoxime
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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 : ~83.33 mg/mL (~282.22 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (7.04 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 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. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.3867 mL | 16.9337 mL | 33.8673 mL | |
| 5 mM | 0.6773 mL | 3.3867 mL | 6.7735 mL | |
| 10 mM | 0.3387 mL | 1.6934 mL | 3.3867 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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