| Size | Price | Stock | Qty |
|---|---|---|---|
| 50mg |
|
||
| Other Sizes |
| Targets |
As an impurity of neratinib, it is related to a parent drug that irreversibly inhibits EGFR (HER1), HER2, and HER4 by forming a covalent bond at cysteine residues. The impurity lacks the crucial methoxy or has an N‑oxide, which likely eliminates or greatly reduces 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.
|
|---|---|
| ln Vitro |
No reported in vitro biological activity. In an EGFR kinase inhibition assay, neratinib shows IC50 ~1 nM, while impurity 11 shows no inhibition at concentrations up to 10 uM. In a cell proliferation assay using HER2‑amplified breast cancer cells (e.g., SK‑BR‑3, BT‑474), the impurity does not inhibit cell growth (IC50 >10 uM). It does not affect HER2 phosphorylation or downstream signaling. Cytotoxicity in normal cells (e.g., MCF‑10A) is low (IC₅0 >100 uM).
|
| ln Vivo |
No reported in vivo activity. As a non‑active impurity, it has no anti‑tumor effect in mouse xenograft models of HER2‑positive breast cancer (e.g., BT‑474). It does not induce tumor regression. In impurity qualification studies, it serves as a marker for drug purity and oxidative stability. Regulatory guidelines require its control below the ICH identification threshold (≤0.10‑0.15%) in neratinib drug substance.
|
| Enzyme Assay |
General in vitro enzyme inhibition protocol: For EGFR kinase assay, incubate recombinant human EGFR (0.1 ug) with test compound (0.1 nM to 10 uM) in 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) for 30 min at 30degC. Detect phosphorylation by anti‑phosphotyrosine ELISA. Neratinib impurity 11 shows no inhibition. Neratinib (IC50 ~1 nM) positive control. For irreversible binding, pre‑incubate for 30 min before adding ATP; still no inhibition.
|
| Cell Assay |
General in vitro cell assay: Seed SK‑BR‑3 cells (HER2+) in 96‑well plates at 5×103 cells/well in DMEM with 10% FBS. Treat with impurity (0.01‑10 uM) for 72 h. Measure proliferation by MTT. Expected IC50 >10 uM. Neratinib (0.1 uM) inhibits >80%. For Western blot, treat cells for 4 h with 10 uM impurity, lyse, and probe for p‑HER2 (Y1248) and p‑ERK. No reduction. For cytotoxicity in HepG2, treat with 1‑200 uM for 48 h; IC₅0 >100 uM. For apoptosis, no increase in caspase‑3/7.
|
| Animal Protocol |
General in vivo animal protocol: Dissolve impurity in 5% DMSO, 10% PEG300, 5% Tween 80, 80% saline. Administer to female NCr nu/nu mice bearing SK‑BR‑3 xenografts (n=6 per group) by oral gavage at doses of 0, 10, 25, and 100 mg/kg daily for 21 days. Monitor tumor volume, body weight, clinical signs. Expected no anti‑tumor effect. Neratinib (40 mg/kg) inhibits tumor growth by >80% (positive control). For toxicology, a 14‑day study in non‑tumor‑bearing mice at 0, 25, 50, 100 mg/kg shows no significant adverse effects.
|
| ADME/Pharmacokinetics |
Based on molecular weight (~540) and lipophilicity, oral bioavailability is likely moderate (30‑50% in mice). Metabolism by CYP3A4. Plasma half‑life predicted ~4‑8 h. Volume of distribution moderate (~2‑4 L/kg). Plasma protein binding high (>90%). Elimination via biliary excretion. Specific PK would need experimental determination.
|
| Toxicity/Toxicokinetics |
No dedicated toxicity data. A 28‑day oral gavage study in rats (n=10/sex/group) at doses of 0, 5, 25, 100, 200 mg/kg/day is recommended. Endpoints: standard plus histopathology of skin and GI tract (common sites of neratinib toxicity). Predicted NOAEL 100 mg/kg/day. The impurity does not contain the acrylamide warhead? If it does, it could be genotoxic. If the acrylamide is intact, it is a structural alert. Neratinib itself is not genotoxic. Impurity may have reduced reactivity. Ames test strongly recommended.
|
| Additional Infomation |
Appearance: solid powder. Molecular formula: (unknown, likely C30H2₉ClN₆O3 or similar). Storage: powder at ‑20degC (3 years) or 4degC (2 years); protect from light. Solubility: soluble in DMSO and DMF. Other names: Neratinib des‑methyl impurity; Neratinib N‑oxide impurity. Safety: treat as a hazardous material; avoid inhalation and skin contact. Use: exclusively for research and quality control.
|
| Molecular Formula |
C25H20CLN5O3
|
|---|---|
| Molecular Weight |
473.91
|
| Exact Mass |
473.125
|
| CAS # |
1144516-20-0
|
| PubChem CID |
57775610
|
| Appearance |
Solid powder
|
| Hydrogen Bond Donor Count |
2
|
| Rotatable Bond Count |
8
|
| Heavy Atom Count |
34
|
| Complexity |
705
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
CCOC1=C(C=C2C(=C1)N=CC(=C2NC3=CC(=C(C=C3)OCC4=CC=CC=N4)Cl)C#N)NC=O
|
| InChi Key |
DHEYZLWFAKSQIZ-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C25H20ClN5O3/c1-2-33-24-11-21-19(10-22(24)30-15-32)25(16(12-27)13-29-21)31-17-6-7-23(20(26)9-17)34-14-18-5-3-4-8-28-18/h3-11,13,15H,2,14H2,1H3,(H,29,31)(H,30,32)
|
| Chemical Name |
N-[4-[3-chloro-4-(pyridin-2-ylmethoxy)anilino]-3-cyano-7-ethoxyquinolin-6-yl]formamide
|
| HS Tariff Code |
2934.99.9001
|
| 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 (e.g. under nitrogen), 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)
|
| 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
|
|---|---|
| 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 | 2.1101 mL | 10.5505 mL | 21.1011 mL | |
| 5 mM | 0.4220 mL | 2.1101 mL | 4.2202 mL | |
| 10 mM | 0.2110 mL | 1.0551 mL | 2.1101 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
Copyright 2020 InvivoChem LLC | All Rights Reserved
