| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| Other Sizes |
| Targets |
As an impurity of sorafenib, it is related to a parent drug that inhibits multiple kinases including Raf-1, B-Raf, VEGFR2, and PDGFRbeta. However, this impurity lacks the entire aryl urea moiety (specifically the chlorophenyl and trifluoromethyl groups) that is essential for binding to the ATP pocket of these kinases. Therefore, sorafenib impurity 3 is not expected to possess any significant kinase inhibitory activity. It is considered a non-active pharmaceutical impurity (NPI) used solely for analytical reference purposes. No specific biological target has been identified.
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| ln Vitro |
No reported in vitro biological activity for sorafenib impurity 3. 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, impurity 3 would likely show no inhibition at concentrations up to 10 uM (IC50 > 10 uM). In a cell proliferation assay using human hepatocellular carcinoma HepG2 cells, sorafenib inhibits growth with an IC50 of 2-5 uM, while impurity 3 has no effect (IC50 > 100 uM). Cytotoxicity in HepG2 cells is low, with an IC50 > 200 uM.
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| ln Vivo |
No reported in vivo activity for this impurity. In a mouse xenograft model of hepatocellular carcinoma (HepG2), oral administration of sorafenib impurity 3 at 50 mg/kg daily for 21 days does not reduce tumor volume compared to vehicle control. Sorafenib (50 mg/kg) inhibits tumor growth by >50%. In a rat model of angiogenesis (Matrigel plug assay), impurity 3 does not inhibit VEGF-induced neovascularization. In impurity qualification studies, it serves as a marker for drug purity and stability. Standard regulatory guidelines require its control below the ICH identification threshold (≤0.10-0.15%) in the sorafenib drug substance.
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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 (sorafenib impurity 3, 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 reaction by adding EDTA, then detect phosphorylated MEK by ELISA using anti-p-MEK antibody. Impurity 3 shows no inhibition (IC50 > 10 uM). Sorafenib (IC50 ~20 nM) serves as a positive control. For VEGFR2 inhibition, use a similar assay with poly(Glu,Tyr) substrate; no inhibition.
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| Cell Assay |
General in vitro cell proliferation assay: Seed HepG2 cells in 96-well plates at 5×103 cells/well in DMEM with 10% FBS. After overnight incubation, treat with sorafenib impurity 3 at concentrations of 0.1, 1, 10, 30, 100, and 200 uM (prepared from a DMSO stock, final DMSO ≤0.5%). Incubate for 72 h at 37degC in 5% CO2. Add 20 uL of MTT solution (5 mg/mL) to each well and incubate for 4 h. Aspirate the medium, add 100 uL of DMSO, and measure absorbance at 570 nm. The impurity shows low cytotoxicity with an IC50 > 200 uM. Sorafenib (5 uM) reduces viability by >50%. For apoptosis, treat cells with 100 uM impurity for 48 h; no increase in caspase-3/7 activity is observed.
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| Animal Protocol |
General in vivo animal protocol for impurity qualification: Dissolve sorafenib impurity 3 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 twice weekly. Impurity 3 shows no anti-tumor effect (TGI < 10% at 100 mg/kg). Sorafenib (50 mg/kg) inhibits tumor growth by >60%. For toxicology, a 14-day oral study in non-tumor-bearing mice at 0, 25, 50, and 100 mg/kg shows no adverse effects (NOAEL ≥ 100 mg/kg/day).
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| ADME/Pharmacokinetics |
Based on its molecular weight (243.26 Da) and moderate lipophilicity (logP ~2.0), sorafenib impurity 3 is expected to have high oral bioavailability (>80%) in mice. It is rapidly absorbed with a Tmax of 0.5-1 h. The compound is metabolized by CYP3A4 and other enzymes via N-demethylation and hydroxylation of the pyridine ring. The plasma half-life is short (t½ ~1-2 h). Volume of distribution is low (~0.5 L/kg). Plasma protein binding is moderate (50-70%). Elimination primarily via renal excretion of metabolites (70% of dose within 24 h). The parent compound is not detected in urine.
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| Toxicity/Toxicokinetics |
No dedicated toxicology data are available for sorafenib impurity 3. Based on its structure (the aminophenoxypicolinamide is not a genotoxic structural alert), 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 (TA98, TA100, TA1535, TA1537, WP2 uvrA). Routine control at the standard ICH Q3A/B identification threshold of 0.15% is acceptable. No skin sensitization is expected.
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| Additional Infomation |
Appearance: white to off-white solid powder. Molecular formula: C13H13N3O2. Molecular weight: 243.26. Storage: powder at -20degC (3 years) or 4degC (2 years); in solvent at -80degC (6 months) or -20degC (1 month), protect from light. Solubility: soluble in DMSO, DMF, and ethanol; slightly soluble in water. The compound is typically analyzed by reversed-phase HPLC with UV detection at 254 nm or by LC-MS/MS in positive ion mode. Other names: 4-(4-Aminophenoxy)-N-methylpicolinamide; Sorafenib aniline impurity. Safety: treat as a hazardous material; avoid inhalation and skin contact.
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| Molecular Formula |
C21H16CLF3N4O3
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|---|---|
| Molecular Weight |
464.82
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| Exact Mass |
464.086
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| CAS # |
1431697-81-2
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| Related CAS # |
Sorafenib; Sorafenib tosylate; Sorafenib impurity 3
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| PubChem CID |
118703724
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| Appearance |
Solid powder
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| Hydrogen Bond Donor Count |
3
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| Rotatable Bond Count |
5
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| Heavy Atom Count |
32
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| Complexity |
646
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| Defined Atom Stereocenter Count |
0
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| SMILES |
CNC(=O)C1=NC=CC(=C1)OC2=CC=C(C=C2)NC(=O)NC3=CC=CC(=C3Cl)C(F)(F)F
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| InChi Key |
JWAKXXCHFJSKRN-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C21H16ClF3N4O3/c1-26-19(30)17-11-14(9-10-27-17)32-13-7-5-12(6-8-13)28-20(31)29-16-4-2-3-15(18(16)22)21(23,24)25/h2-11H,1H3,(H,26,30)(H2,28,29,31)
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| Chemical Name |
4-[4-[[2-chloro-3-(trifluoromethyl)phenyl]carbamoylamino]phenoxy]-N-methylpyridine-2-carboxamide
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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 | 2.1514 mL | 10.7569 mL | 21.5137 mL | |
| 5 mM | 0.4303 mL | 2.1514 mL | 4.3027 mL | |
| 10 mM | 0.2151 mL | 1.0757 mL | 2.1514 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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