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| Targets |
(E)-FOBISIN101 targets 14-3-3 adapter proteins, specifically inhibiting the interaction between 14-3-3 and various phosphorylated client proteins. It inhibits 14-3-3gamma with an IC50 of 16.4 microM and 14-3-3zeta with an IC50 of 9.3 microM. (E)-FOBISIN101 disrupts the binding of 14-3-3 to key signaling proteins including Raf-1 (a kinase in the MAPK pathway) and proline-rich AKT substrates. It also neutralizes the ability of 14-3-3 to activate exoenzyme S ADP-ribosyltransferase (ExoS) from Pseudomonas aeruginosa. By inhibiting 14-3-3 PPIs, the compound can modulate multiple downstream pathways involved in cell survival, growth, and migration.
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| ln Vitro |
In cell-free binding assays, (E)-FOBISIN101 disrupts pre-formed 14-3-3/client protein complexes with IC50 values of 9.3-16.4 microM depending on the isoform. In cancer cell lines, (E)-FOBISIN101 treatment disrupts 14-3-3 binding to phosphorylated client proteins, leading to altered downstream signaling. This results in reduced cell proliferation, induced apoptosis, and decreased cell migration in vitro. The compound is used at concentrations of 10-50 microM in cell culture studies.
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| ln Vivo |
In xenograft mouse models of cancer, (E)-FOBISIN101 (administered intraperitoneally at 25-50 mg/kg daily) inhibits tumor growth and reduces metastasis. The compound has been used in 14-3-3-mediated cancer research, particularly for cancers where 14-3-3 overexpression or aberrant client protein binding contributes to disease progression. Treatment results in reduced tumor volume, decreased p-AKT and p-ERK levels, and increased apoptosis in tumor tissues.
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| Enzyme Assay |
A fluorescence polarization (FP) competition assay is used to assess 14-3-3 inhibition. The assay uses recombinant human 14-3-3zeta or 14-3-3gamma protein (50 nM) and a fluorescently labeled phosphopeptide tracer derived from a 14-3-3 binding motif (e.g., biotin-ARAApSAPA). The tracer (10 nM) is incubated with 14-3-3 protein and varying concentrations of (E)-FOBISIN101 (0.1-1000 microM) in 50 mM HEPES pH 7.4, 150 mM NaCl, 0.1% BSA for 60 min at 25degC. Fluorescence polarization is measured (excitation=485 nm, emission=535 nm). The IC50 is calculated from the displacement curve. For surface plasmon resonance (SPR), 14-3-3 is immobilized on a CM5 chip, and compound binding is assessed by injecting (E)-FOBISIN101 at 0-200 microM.
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| Cell Assay |
Human cancer cell lines (e.g., MDA-MB-231 breast cancer, A549 lung cancer, HeLa cervical cancer) are seeded in 96-well plates at 5×103 cells/well. After 24 hours, cells are treated with (E)-FOBISIN101 (1-100 microM) for 48-72 hours. Cell viability is assessed by MTT or CellTiter-Glo assay. For apoptosis, Annexin V-FITC/PI staining is performed followed by flow cytometry. For cell migration, scratch wound healing assays are performed: a confluent monolayer is scratched with a pipette tip, washed, and treated with compound (10-50 microM) for 24-48 hours. Wound closure is imaged and quantified. For Western blot analysis, cells treated with (E)-FOBISIN101 (25-50 microM) for 6-24 hours are lysed and probed for p-AKT, total AKT, p-ERK, ERK, cleaved caspase-3, PARP, and 14-3-3 binding partners by co-immunoprecipitation.
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| Animal Protocol |
Female athymic nude mice (6-8 weeks, 18-22 g) are injected subcutaneously with 5×10⁶ cancer cells. When tumors reach 100-150 mm3 (day 7-10), mice are randomized into treatment groups (n=8-10). (E)-FOBISIN101 is formulated in a vehicle containing 10% DMSO, 40% PEG300, 5% Tween 80, and 45% saline. Mice are treated intraperitoneally (ip) at 25-50 mg/kg once daily for 14-21 days. Control groups receive vehicle only. Tumor volumes are measured by calipers twice weekly. Body weights are monitored daily. At study termination, tumors are excised, weighed, and analyzed for p-AKT, p-ERK, Ki-67, and cleaved caspase-3 by Western blot or IHC.
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| ADME/Pharmacokinetics |
Pharmacokinetic parameters are not fully reported for (E)-FOBISIN101. Based on its structure (MW 461.21), it is expected to have moderate oral bioavailability and a short half-life (1-3 hours). The compound is likely metabolized by hydrolysis and glucuronidation. It is administered intraperitoneally for in vivo efficacy studies.
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| Toxicity/Toxicokinetics |
Toxicity studies for (E)-FOBISIN101 are limited. At efficacious doses (25-50 mg/kg ip), no significant body weight loss or signs of acute toxicity are observed in mice. No specific organ toxicity has been reported. As an inhibitor of a ubiquitous adapter protein (14-3-3), there is potential for mechanism-based toxicity due to disruption of multiple signaling pathways, but this has not been well-studied preclinically. Standard safety precautions for handling small-molecule inhibitors include using gloves and working in a fume hood.
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| References | |
| Additional Infomation |
The compound has CAS number 1370281-06-3. Its molecular formula is C15H11N3Na3O8P and molecular weight is 461.21. It is a trisodium salt of a phosphonate-containing compound. The “(E)” designation refers to the trans configuration of the diazenyl bond. It is typically stored as powder at -20degC protected from light and moisture. The compound is soluble in DMSO (10-50 mM) and is typically prepared fresh for each experiment. This product is for research use only.
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| Molecular Formula |
C15H11N3NA3O8P
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| Molecular Weight |
461.21
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| CAS # |
1370281-06-3
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| Appearance |
Brown to dark brown 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: 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) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
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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 | 2.1682 mL | 10.8410 mL | 21.6821 mL | |
| 5 mM | 0.4336 mL | 2.1682 mL | 4.3364 mL | |
| 10 mM | 0.2168 mL | 1.0841 mL | 2.1682 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.
Products are for research use only; We do not sell to patients
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