| Size | Price | Stock | Qty |
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| Targets |
IGF1R, CDK2/CycE, VEGFR-2, Src kinase, and the Stat3 pathway. By inhibiting these targets, E804 disrupts multiple hallmarks of cancer, including cell cycle progression, survival signaling, and new blood vessel formation. Its ability to block constitutive Stat3 activation is particularly relevant in breast and prostate cancers.
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| ln Vitro |
Water-soluble isatin derivative indirubin derivative E804 has an IC50 of 0.65 μM and is a strong inhibitor of the insulin-like growth factor 1 receptor (IGF1R). Additionally, CDK2/CycE is inhibited by Indirubin Derivative E804 with an EC50 of 0.23 μM[1].
In vitro, E804 potently inhibits IGF1R autophosphorylation and downstream Akt/ERK activation. It also inhibits CDK2 activity, causing G1/S arrest. It blocks VEGF-induced VEGFR-2 phosphorylation and endothelial tube formation. In cancer cell lines, it reduces Stat3 phosphorylation and downregulates anti-apoptotic genes, leading to caspase-dependent apoptosis. IC50 values vary by cell line, typically in sub-micromolar range. |
| ln Vivo |
In vivo, E804 has shown tumor growth inhibition in mouse xenograft models of breast and prostate cancer when administered orally or intraperitoneally. Tumor microvessel density is reduced, indicating anti-angiogenic effects. Biomarker analyses confirm target inhibition in tumor tissues. However, detailed pharmacokinetic and dose-response data are limited in public literature; further studies are needed to establish optimal dosing and efficacy schedules.
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| Enzyme Assay |
Cell-free kinase assays use purified recombinant kinases (IGF1R, CDK2/CycE, VEGFR-2, Src). Reactions contain ATP, peptide substrate, and varying E804 concentrations. Phosphorylation is measured by radiometric (33P-ATP) or fluorescence-based (ADP-Glo) methods. IC50 values are derived from dose-response curves. For Stat3 inhibition, a DNA-binding ELISA or electrophoretic mobility shift assay can be used with purified Stat3 protein.
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| Cell Assay |
Cancer cells (e.g., MCF-7, PC-3, HCT116) are treated with 0.1–10 µM E804 for 24–72 h. Cell viability is measured by MTT or SRB. Apoptosis is quantified by Annexin V/PI flow cytometry and caspase-3/7 activity. Western blotting assesses phospho-IGF1R, phospho-Stat3, and downstream targets. Cell cycle analysis is performed by propidium iodide staining. The anti-angiogenic effect is evaluated in HUVEC tube formation assays.
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| Animal Protocol |
In vivo efficacy studies use female athymic nude mice bearing subcutaneous xenografts of breast or prostate cancer. When tumors reach ~100 mm³, mice receive E804 orally (e.g., 50 mg/kg) daily for 2–3 weeks. Tumor volume is measured twice weekly. At study end, tumors are excised for immunohistochemistry (CD31 for microvessels, phospho-Stat3, Ki67) and Western blotting. Plasma drug levels may be analyzed by LC-MS/MS for pharmacokinetic correlation.
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| ADME/Pharmacokinetics |
Molecular formula C20H19N3O4, MW 365.38. Appearance: brown to reddish-brown solid. Solubility: DMSO ~125 mg/mL (~342 mM). For in vivo, formulations may include 10% DMSO + 40% PEG300 + 5% Tween-80 + 45% saline. Storage: powder at -20°C for 3 years; solution at -80°C for 1 year. Purity ≥98% by HPLC.
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| Toxicity/Toxicokinetics |
No comprehensive toxicology data are publicly available. As a multi-kinase inhibitor, it may have off-target effects on normal tissues. In vitro cytotoxicity in non-malignant cells (e.g., MCF-10A) should be assessed to determine selectivity. Standard genotoxicity and cardiovascular safety assays (hERG) are recommended before further development. In animal studies, weight loss and gastrointestinal disturbances may occur at high doses.
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| References | |
| Additional Infomation |
E804 is a research compound, not approved for human use. It is a valuable pharmacological probe for studying IGF1R, CDK2, and Stat3 signaling in cancer. It is more water-soluble than parent indirubin, facilitating formulation. It has been used in combination with other targeted agents in preclinical models. No clinical trials have been reported. Its multi-targeted profile makes it useful for investigating cross-talk between growth factor and cytokine pathways.
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| Molecular Formula |
C20H19N3O4
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| Molecular Weight |
365.382564783096
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| Exact Mass |
365.137
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| CAS # |
854171-35-0
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| PubChem CID |
135398511
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| Appearance |
Brown to reddish brown solid powder
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| LogP |
2.4
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| Hydrogen Bond Donor Count |
4
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| Hydrogen Bond Acceptor Count |
6
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| Rotatable Bond Count |
6
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| Heavy Atom Count |
27
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| Complexity |
582
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| Defined Atom Stereocenter Count |
0
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| SMILES |
O(CCC(CO)O)/N=C1/C2C=CC=CC=2N=C/1C1=C(NC2C=CC=CC1=2)O
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| InChi Key |
TWOSIFOFWWXXIG-PTGBLXJZSA-N
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| InChi Code |
InChI=1S/C20H19N3O4/c24-11-12(25)9-10-27-23-18-14-6-2-4-8-16(14)21-19(18)17-13-5-1-3-7-15(13)22-20(17)26/h1-8,12,22,24-26H,9-11H2/b23-18+
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| Chemical Name |
4-[(E)-[2-(2-hydroxy-1H-indol-3-yl)indol-3-ylidene]amino]oxybutane-1,2-diol
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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 : ~125 mg/mL (~342.11 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (5.69 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 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. Solubility in Formulation 2: 2.08 mg/mL (5.69 mM) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication. 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 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline 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 | 2.7369 mL | 13.6844 mL | 27.3688 mL | |
| 5 mM | 0.5474 mL | 2.7369 mL | 5.4738 mL | |
| 10 mM | 0.2737 mL | 1.3684 mL | 2.7369 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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