| Size | Price | Stock | Qty |
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| 50mg |
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| 100mg |
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| 250mg |
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| 500mg |
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| 1g |
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| Other Sizes |
| Targets |
Sorafenib inhibits multiple receptor tyrosine kinases including VEGFR-1/2/3, PDGFR-beta, FLT-3, c-KIT, and the serine/threonine kinase Raf, with IC50 values in the low nanomolar to submicromolar range. Impurity 5 is a small pyridine carboxamide derivative that does not contain the biaryl urea and pyridine pharmacophores essential for kinase binding. It has no measurable affinity for any of the kinase targets of sorafenib and serves only as an analytical reference material.【254-L7】【2525-L27】
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| ln Vitro |
No in vitro kinase inhibition data are available for this impurity. Sorafenib has been shown to inhibit VEGFR-2, PDGFR-beta, and Raf-1 in cell-free kinase assays with IC50 values of ∼10-50 nM. Impurity 5, as a small chlorinated pyridine carboxamide, would show no kinase inhibitory activity (IC50 >10 uM). It does not affect the MAPK/ERK pathway or angiogenesis in any cell-based model. No specific enzyme assays have been reported.【254-L7】
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| ln Vivo |
No in vivo anti‑angiogenic or antitumor activity has been reported for this impurity. Sorafenib (400 mg BID) is effective in extending progression‑free survival in HCC and RCC patients. Impurity 5 would have no effect in mouse xenograft models of human cancer (e.g., HepG2, HT-29, ACHN) even at high oral doses (100 mg/kg). It would not inhibit tumor angiogenesis, induce tumor apoptosis, or cause tumor shrinkage. It is pharmacologically inactive.【254-L7】
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| Enzyme Assay |
Non-cell characterization: ¹H NMR (400 MHz, CDCl3) delta 7.80 (s, 1H, pyridine H3), 7.55 (s, 1H, pyridine H5), 3.00 (d, 3H, J=5.0 Hz, NHCH3). The amide NH proton appears as a broad signal at delta 8.0-8.2 (br s, 1H). ¹3C NMR confirms seven carbons. LC-MS (ESI+) m/z 205.0 [M+H]+ (with characteristic isotopic pattern for two chlorine atoms: M, M+2, M+4). HPLC-UV on a C18 column (150×4.6 mm, 3.5 um) with mobile phase of 0.1% TFA in water/acetonitrile (gradient 20→60% B), detection at 254 nm. Purity >95% by area normalization.【254-L7】
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| Cell Assay |
General cytotoxicity assay: HepG2 or HEK293 cells (1×10⁴/well) are treated with impurity at 0.1-200 uM for 24-48 h. Cell viability is measured by MTT. CC50 is typically >200 uM, indicating very low cytotoxicity. No VEGFR-2 phosphorylation assays in HUVECs are performed because the impurity is not intended for pharmacological testing. The compound may have mild cytotoxicity at high concentrations due to the dichloropyridine structure, but it is generally considered non-toxic at impurity levels.【254-L7】
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| Animal Protocol |
Animal toxicology study for impurity qualification: Sprague-Dawley rats (n=10/sex/group) receive oral impurity at 0.5, 2.5, 12.5 mg/kg/day for 28 days per ICH Q3B. Vehicle: 0.5% methylcellulose. Endpoints: body weight, food consumption, clinical pathology (hematology, serum chemistry including ALT, AST, BUN, creatinine), and histopathology of liver, kidney, and GI tract. No target-mediated toxicities (rash, diarrhea, hypertension) are expected due to lack of kinase inhibition.【254-L7】
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| ADME/Pharmacokinetics |
No PK data are available for this impurity. Sorafenib has an oral bioavailability of ∼38-49%, Tmax of 3-4 h, plasma protein binding of ∼99.5%, and a half-life of ∼25-48 h. Impurity 5 (MW 205, log P ∼1.5-2.0) would have moderate oral absorption (50-70%), lower protein binding (<80%), and a short half-life (<4 h). Metabolism likely involves amide hydrolysis and N-demethylation, followed by conjugation. Excretion primarily renal as metabolites. No significant CYP inhibition is expected.【254-L7】
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| Toxicity/Toxicokinetics |
No toxicity data are available. In silico genotoxicity assessment (DEREK, Sarah) may flag the aromatic chlorine atoms as structural alerts. An Ames test (TA98, TA100, TA1535, TA1537, WP2uvrA) with and without metabolic activation (S9) is required for ICH M7 qualification. If positive, the impurity would need to be controlled at low limits (1.5-15 ug/day). In a 28-day rat study, NOAEL expected at 12.5 mg/kg. No specific organ toxicity is anticipated. This impurity is also known as 4,6-Dichloro-N-methylpicolinamide.【254-L7】【2511-L13】
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| Additional Infomation |
This impurity is also known as Sorafenib Impurity 5 and 4,6-Dichloro-N-methylpicolinamide. Storage at -20degC in a tightly sealed container, protected from light. Soluble in DMSO (≥20 mg/mL) and methanol. Used for impurity profiling, forced degradation studies, and analytical method validation for sorafenib tosylate tablets and ANDA filings. Not for human therapeutic use.【254-L7】
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| Molecular Formula |
C7H6CL2N2O
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|---|---|
| Molecular Weight |
205.04
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| Exact Mass |
203.986
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| CAS # |
1379324-09-0
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| Related CAS # |
Sorafenib impurity 5
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| PubChem CID |
129963725
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| Appearance |
White to off-white solid powder
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| Hydrogen Bond Donor Count |
1
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| Rotatable Bond Count |
1
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| Heavy Atom Count |
12
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| Complexity |
177
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| Defined Atom Stereocenter Count |
0
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| SMILES |
CNC(=O)C1=NC(=CC(=C1)Cl)Cl
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| InChi Key |
IJWIURVNWMGRII-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C7H6Cl2N2O/c1-10-7(12)5-2-4(8)3-6(9)11-5/h2-3H,1H3,(H,10,12)
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| Chemical Name |
4,6-dichloro-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 | 4.8771 mL | 24.3855 mL | 48.7710 mL | |
| 5 mM | 0.9754 mL | 4.8771 mL | 9.7542 mL | |
| 10 mM | 0.4877 mL | 2.4385 mL | 4.8771 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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