| Size | Price | Stock | Qty |
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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 |
Nintedanib potently inhibits VEGFR-1/2/3, FGFR-1/2/3, and PDGFR-alpha/beta tyrosine kinases with IC50 values of ∼10-100 nM. Impurity 41 is a simple nitroaniline derivative that does not contain the oxindole or indolinone pharmacophore required for kinase binding. It shows no measurable affinity for VEGFR, FGFR, or PDGFR and serves only as an analytical reference material.
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|---|---|
| ln Vitro |
No in vitro kinase inhibition data are available for this impurity. Nintedanib has been shown to inhibit VEGFR-2 with IC50 ∼10 nM in cell-free kinase assays. Impurity 41, as a N-methyl-N-(4-nitrophenyl)acetamide derivative, would show no activity against VEGFR, FGFR, or PDGFR even at concentrations up to 100 uM. No specific enzyme assays have been performed.
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| ln Vivo |
No in vivo anti-fibrotic or anti-angiogenic activity has been reported for this impurity. Nintedanib (150 mg BID) reduces the decline in lung function in IPF patients. Impurity 41 would have no effect in the bleomycin-induced pulmonary fibrosis mouse model at doses up to 100 mg/kg p.o. It does not inhibit VEGF-induced angiogenesis in the Matrigel plug assay.
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| Enzyme Assay |
Non-cell characterization: ¹H NMR (400 MHz, CDCl3) delta 8.25 (d, 2H, J=9.0 Hz, Ar-H ortho to NO2), 7.45 (d, 2H, J=9.0 Hz, Ar-H meta to NO2), 3.40 (s, 3H, NCH3), 2.25 (s, 3H, COCH3). ¹3C NMR confirms nine carbons. LC-MS (ESI+) m/z 195.1 [M+H]+, (ESI-) m/z 193.1 [M-H]-. HPLC-UV on C18 column with mobile phase of 0.1% TFA in water/acetonitrile (gradient 30→70% B), detection 254 nm. Purity >95% by area normalization.
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| Cell Assay |
General cytotoxicity assay: HEK293 or HepG2 cells (1×10⁴/well) are treated with impurity at 0.1-200 uM for 24-48 h. Cell viability is measured by MTT assay. CC50 is typically >200 uM, indicating very low cytotoxicity. No VEGFR phosphorylation assays in HUVECs are performed because the impurity is not intended for pharmacological testing.
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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), blood pressure, and histopathology of liver, kidney, and blood vessels.
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| ADME/Pharmacokinetics |
No PK data are available for this impurity. Nintedanib has an oral bioavailability of ∼60%, Tmax of 2-4 h, plasma protein binding of ∼98%, and a half-life of ∼10 h. Impurity 41 (MW 194, log P ∼1.5) would have moderate oral absorption (40-60%). Metabolism likely involves reduction of the nitro group to an amine followed by acetylation. t½ predicted ∼3-5 h. Excretion primarily renal.
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| Toxicity/Toxicokinetics |
No toxicity data are available for this impurity. In silico genotoxicity assessment (DEREK, Sarah) is required. The nitroaniline group is a known structural alert for mutagenicity. An Ames test (five strains, +/-S9) is mandatory for ICH M7 qualification due to the potential for nitro reduction to reactive intermediates. If positive, the impurity would need to be controlled at low levels (1.5-15 ug/day). NOAEL expected at 2.5 mg/kg in 28-day rat study.
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| Additional Infomation |
This impurity is also known as N-Methyl-N-(4-nitrophenyl)acetamide and Nintedanib Impurity 41. Storage at 4degC in a tightly sealed container, protected from light. Soluble in DMSO (≥20 mg/mL), ethanol, and dichloromethane. Used as a reference standard for related substance testing in nintedanib esylate capsules and ANDA filings.
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| Molecular Formula |
C9H10N2O3
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|---|---|
| Molecular Weight |
194.19
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| Exact Mass |
194.069
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| CAS # |
121-95-9
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| Related CAS # |
Nintedanib impurity 41
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| PubChem CID |
67143
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| Appearance |
Solid powder
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| Hydrogen Bond Donor Count |
0
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| Rotatable Bond Count |
1
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| Heavy Atom Count |
14
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| Complexity |
229
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| Defined Atom Stereocenter Count |
0
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| SMILES |
CC(=O)N(C)C1=CC=C(C=C1)[N+](=O)[O-]
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| InChi Key |
DKZFYTVXALXRSH-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C9H10N2O3/c1-7(12)10(2)8-3-5-9(6-4-8)11(13)14/h3-6H,1-2H3
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| Chemical Name |
N-methyl-N-(4-nitrophenyl)acetamide
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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 | 5.1496 mL | 25.7480 mL | 51.4960 mL | |
| 5 mM | 1.0299 mL | 5.1496 mL | 10.2992 mL | |
| 10 mM | 0.5150 mL | 2.5748 mL | 5.1496 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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