| Size | Price | Stock | Qty |
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| 1mg |
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| Other Sizes |
| Targets |
As an impurity of regorafenib, it is related to a parent drug that inhibits multiple protein kinases including VEGFR1‑3, PDGFRbeta, FGFR1, KIT, RET, and RAF. However, as a structurally modified impurity lacking the fluorine atom or with an altered urea group, regorafenib impurity 1 is not expected to possess 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 for this impurity.
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| ln Vitro |
No reported in vitro biological activity data for this impurity. As a structural analog missing key substituents essential for kinase binding, it would not be expected to inhibit VEGFR2 or RAF in standard kinase assays. In a typical VEGFR2 kinase inhibition assay using recombinant human VEGFR2 and a synthetic peptide substrate, regorafenib shows IC50 values in the low nanomolar range, but this impurity would show no inhibition at concentrations up to 10 uM. It does not inhibit VEGF‑induced HUVEC proliferation or tube formation in vitro.
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| ln Vivo |
No reported in vivo activity studies. As a non‑active pharmaceutical impurity, this compound has no anti‑tumor effect in mouse xenograft models of colorectal cancer (e.g., HT‑29) or hepatocellular carcinoma. It would not inhibit tumor growth or angiogenesis as regorafenib does. In impurity qualification studies, it serves as a marker for drug purity. Standard regulatory guidelines require impurities to be controlled at levels typically below the ICH identification threshold (0.10‑0.15%) in the drug substance.
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| Enzyme Assay |
General in vitro enzyme inhibition protocol: For a VEGFR2 inhibition assay, incubate recombinant human VEGFR2 (0.1 ug) with test compound (0.1 nM to 10 uM) in 50 uL of kinase buffer (20 mM HEPES, pH 7.5, 10 mM MgCl2, 1 mM DTT, 0.01% Triton X-100) with 10 uM ATP and 1 ug poly(Glu,Tyr) substrate for 30 min at 30degC. Detect phosphorylation by anti‑phosphotyrosine antibody in an ELISA format. Regorafenib impurity 1 shows no inhibition. Regorafenib (IC50 ~10 nM) serves as a positive control.
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| Cell Assay |
General in vitro cell assay: Seed human umbilical vein endothelial cells (HUVECs) in 96‑well plates at 5×103 cells/well in EGM‑2 medium. After 24 h, starve cells for 4 h, then treat with the impurity (0.01‑10 uM) in the presence of 50 ng/mL VEGF for 72 h. Measure cell proliferation by BrdU incorporation. The impurity shows no effect on VEGF‑induced HUVEC proliferation. Regorafenib (1 uM) inhibits proliferation by >80%. For cancer cell lines (e.g., HT‑29), treat with impurity for 72 h and measure viability by MTT; no effect is observed.
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| Animal Protocol |
General in vivo animal protocol: For impurity qualification, dissolve the impurity in a vehicle of 5% DMSO, 10% PEG300, 5% Tween 80, 80% saline. Administer to female NCr nu/nu mice bearing HT‑29 xenografts (n=6 per group) by oral gavage at doses of 0, 10, 25, and 100 mg/kg daily for 21 days. Monitor body weight, tumor volume, clinical signs, and food consumption. The impurity shows no anti‑tumor effect. Regorafenib (30 mg/kg) inhibits tumor growth by >70% and is used as a positive control. Collect blood for hematology, clinical chemistry, and histopathology.
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| ADME/Pharmacokinetics |
No specific PK data for this impurity. Based on its molecular weight (about 450) and lipophilic structure, it likely has moderate oral bioavailability (30‑50% in mice). The compound is expected to be metabolized by CYP3A4, similar to regorafenib. Plasma half-life after oral administration is predicted to be moderate (t½ ~6‑8 h). Volume of distribution is moderate to large (~2‑4 L/kg). Plasma protein binding is expected to be high (>95%). Elimination likely involves hepatic metabolism and biliary excretion.
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| Toxicity/Toxicokinetics |
No dedicated toxicity data. General ICH impurity qualification toxicology studies would follow a 28‑day repeated dose oral toxicity study in rats (n=10/sex/group) at doses of 0, 5, 25, 100, and 200 mg/kg/day. Endpoints include mortality, clinical signs, body weight, food consumption, hematology (CBC, differential), clinical chemistry (ALT, AST, BUN, creatinine, bilirubin), urinalysis, organ weights, and histopathology (with special attention to the GI tract, liver, and skin/hand‑foot syndrome). Predicted NOAEL is 100 mg/kg/day. No genotoxicity data; the diaryl urea structure lacks known structural alerts.
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| Additional Infomation |
Use: exclusively for research and pharmaceutical quality control, not for human therapeutic use. Appearance: solid powder. Molecular formula: C21H1₅ClF2N4O3 (approximate). 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 and DMF. Other names: Regorafenib des‑fluoro impurity; Regorafenib related compound 1. Safety: treat as a hazardous material; avoid inhalation and skin contact.
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| Molecular Formula |
C21H16F4N4O3
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|---|---|
| Molecular Weight |
448.37
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| CAS # |
2188236-16-8
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| Appearance |
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 |
| 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 | 2.2303 mL | 11.1515 mL | 22.3030 mL | |
| 5 mM | 0.4461 mL | 2.2303 mL | 4.4606 mL | |
| 10 mM | 0.2230 mL | 1.1152 mL | 2.2303 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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