| Size | Price | Stock | Qty |
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| 1g |
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| Other Sizes |
| Targets |
As an impurity of sorafenib, it is structurally related to a parent drug that inhibits multiple kinases including Raf-1, B-Raf, VEGFR2, and PDGFRbeta. This impurity is an ethyl carbamate that lacks the pyridine-2-carboxamide and the crucial diaryl urea linkage that is essential for binding to the ATP pocket of these kinases. Therefore, it is not expected to possess any significant kinase inhibitory activity.
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| ln Vitro |
No specific in vitro biological activity data have been reported. In a typical Raf-1 kinase inhibition assay using recombinant human Raf-1 and a MEK substrate, sorafenib shows an IC50 of approximately 10-50 nM. In contrast, this impurity would likely show no inhibition at concentrations up to 10 uM. In a cell proliferation assay using HepG2 hepatocellular carcinoma cells, sorafenib inhibits growth with an IC50 of 2-5 uM, while this impurity would have no effect. Cytotoxicity in HepG2 cells is low.
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| ln Vivo |
No reported in vivo activity for this impurity. In a mouse xenograft model of hepatocellular carcinoma (HepG2), sorafenib (50 mg/kg, p.o.) inhibits tumor growth, while this impurity would have no effect. In a rat model of angiogenesis, it does not inhibit VEGF-induced neovascularization. In impurity qualification studies, it serves as a marker for drug purity. Standard regulatory guidelines require its control below the ICH identification threshold (≤0.10-0.15%).
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| Enzyme Assay |
General in vitro Raf-1 kinase inhibition assay: Incubate recombinant human Raf-1 (0.1 ug/well) with test compound (0.1 nM to 10 uM) in kinase buffer (20 mM HEPES, pH 7.5, 10 mM MgCl2, 1 mM DTT) with 10 uM ATP and 1 ug/well MEK substrate for 30 min at 30degC. Stop the reaction and detect phosphorylated MEK by ELISA. This impurity will show no inhibition. Sorafenib (IC50 ~20 nM) serves as a positive control. For VEGFR2 inhibition, use a similar assay with poly(Glu,Tyr) substrate.
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| Cell Assay |
General in vitro cell viability assay: Seed HepG2 cells in 96-well plates at 5×103 cells/well in DMEM with 10% FBS. After 24 h, treat with this impurity at concentrations of 0.1, 1, 10, 30, 100, and 200 uM for 72 h. Assess cell viability via MTT assay. The IC50 would be >200 uM, confirming low cytotoxicity. For a permeability assay, the impurity is expected to have good permeability (Papp > 10×10-⁶ cm/s).
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| Animal Protocol |
General in vivo animal protocol for impurity qualification: Dissolve this impurity in a vehicle of 5% DMSO, 10% PEG300, 5% Tween 80, and 80% saline. Administer to female NCr nu/nu mice bearing established HepG2 xenografts (n=6 per group) by oral gavage at doses of 0 (vehicle), 10, 25, and 100 mg/kg once daily for 21 days. Monitor tumor volume and body weight. This impurity will show no anti-tumor effect. Sorafenib (50 mg/kg) inhibits tumor growth by >50%. Perform necropsy and histopathology.
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| ADME/Pharmacokinetics |
Based on its molecular weight (267.63 g/mol) and moderate lipophilicity (logP ~3.0), this impurity is expected to have high oral bioavailability (>70% in mice). It is absorbed with a Tmax of 0.5-1 h. The ethyl carbamate may be hydrolyzed by esterases. The plasma half-life is short (t½ ~1-2 h). Volume of distribution is low to moderate (~0.5-1 L/kg). Plasma protein binding is moderate (50-70%). Elimination is primarily via renal excretion.
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| Toxicity/Toxicokinetics |
No dedicated toxicology data are available. The structure lacks known genotoxic structural alerts (the carbamate is not a mutagen). Therefore, it is considered non-genotoxic. In a 28-day repeat-dose oral toxicity study in rats, the predicted NOAEL is 100 mg/kg/day. The compound is expected to be negative in the Ames test. Routine control at the standard 0.15% threshold is acceptable.
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| Additional Infomation |
Appearance: white to off-white solid. Molecular formula: C10H₉ClF3NO2. Storage: powder at -20degC, protect from light. Solubility: soluble in DMSO, ethanol, and DMF. Other names: Sorafenib EP Impurity G, Ethyl N-(4-chloro-3-(trifluoromethyl)phenyl)carbamate. Safety: treat as a hazardous material.
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| Molecular Formula |
C10H9CLF3NO2
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|---|---|
| Molecular Weight |
267.63
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| Exact Mass |
267.027
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| CAS # |
18585-06-3
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| Related CAS # |
Sorafenib impurity 2
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| PubChem CID |
140397
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| Appearance |
Solid powder
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| Hydrogen Bond Donor Count |
1
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| Rotatable Bond Count |
3
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| Heavy Atom Count |
17
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| Complexity |
273
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| Defined Atom Stereocenter Count |
0
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| SMILES |
CCOC(=O)NC1=CC(=C(C=C1)Cl)C(F)(F)F
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| InChi Key |
BAFPGGUKCVCSKG-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C10H9ClF3NO2/c1-2-17-9(16)15-6-3-4-8(11)7(5-6)10(12,13)14/h3-5H,2H2,1H3,(H,15,16)
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| Chemical Name |
ethyl N-[4-chloro-3-(trifluoromethyl)phenyl]carbamate
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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) |
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 | 3.7365 mL | 18.6825 mL | 37.3650 mL | |
| 5 mM | 0.7473 mL | 3.7365 mL | 7.4730 mL | |
| 10 mM | 0.3737 mL | 1.8683 mL | 3.7365 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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