| Size | Price | Stock | Qty |
|---|---|---|---|
| 5mg |
|
||
| 10mg |
|
||
| 50mg |
|
||
| Other Sizes |
| Targets |
Dacomitinib irreversibly and covalently binds to EGFR (ErbB1), HER2 (ErbB2), and HER4 (ErbB4) at the ATP‑binding pocket, forming a covalent bond with Cys797 in EGFR. Dacomitinib impurity 3 is a hydrolyzed derivative of dacomitinib. It may be able to bind reversibly to EGFR but lacks the Michael acceptor (crotonamide) required for covalent binding, or the warhead has been modified. The IC50 for EGFR is likely 100‑fold higher (>0.5 uM).
|
|---|---|
| ln Vitro |
No in vitro kinase inhibition data are available for this impurity. Dacomitinib inhibits wild‑type EGFR with IC50 ∼0.5 nM, HER2 with IC50 ∼1 nM, and HER4 with IC50 ∼10 nM in cell‑free kinase assays. Dacomitinib impurity 3, lacking the reactive crotonamide warhead, would show greatly reduced reversible binding (IC50 >100 nM) and would not form a covalent bond with Cys797. No specific cell‑free kinase assays have been reported for this impurity. No significant cell proliferation inhibition is expected.
|
| ln Vivo |
No in vivo anti‑tumor activity has been reported for this impurity. Dacomitinib (45 mg once daily) is effective in prolonging progression‑free survival in patients with EGFR‑mutant non‑small cell lung cancer. Dacomitinib impurity 3 would have no activity in mouse xenograft models of EGFR‑driven tumors (e.g., NCI‑H1975, PC‑9) even at high oral doses (100 mg/kg). It would not inhibit downstream signaling pathways (MAPK/ERK, PI3K/AKT) or induce apoptosis. It would not cause skin rash or diarrhea (the on‑target side effects of dacomitinib).
|
| Enzyme Assay |
Non‑cell characterization: ¹H NMR (400 MHz, DMSO‑d₆) shows the quinazoline core, the 3‑chloro-4‑fluoroaniline group, the 7‑methoxy group, and the 5‑hydroxypyrrolidin-2-one side chain. Characteristic signals: delta 9.50 (s, 1H, NH of aniline), 8.50 (s, 1H, quinazoline H2), 8.10 (dd, 1H, J=7.0, 2.0 Hz, Ar‑H of aniline), 7.90 (s, 1H, quinazoline H5 or H8), 7.65 (s, 1H, quinazoline H5 or H8), 7.35 (t, 1H, J=9.0 Hz, aniline Ar‑H), 5.65 (d, 1H, J=5.0 Hz, CHOH), 4.90 (m, 1H, CHOH), 4.00 (s, 3H, OCH3), 3.80 (s, 3H, NCH3?), 2.50‑2.70 (m, 2H, pyrrolidinone CH2), 2.20‑2.35 (m, 2H, pyrrolidinone CH2). LC‑MS (ESI+) m/z 403.1 [M+H]+. HPLC‑UV on a C18 column with acetonitrile/0.1% TFA in water (gradient 20→60% B), detection at 254 nm and 340 nm. Purity >95% by area normalization.
|
| Cell Assay |
General cytotoxicity assay: EGFR‑mutant NSCLC cell lines (e.g., HCC827, NCI‑H1975) are seeded in 96‑well plates (5×103/well) and treated with impurity at 0.1‑100 uM for 72 h. Cell viability is measured by CellTiter‑Glo. The IC50 for inhibition of cell proliferation is expected to be >10 uM, much higher than that of dacomitinib (IC50 ∼0.01‑0.1 uM). No EGFR phosphorylation assays (Western blot for p‑EGFR Tyr1068) are performed because the impurity is not intended for pharmacological testing. The CC50 is estimated to be >50 uM.
|
| Animal Protocol |
Animal toxicology study for impurity qualification: Sprague‑Dawley rats (n=10 per sex per group) receive oral Dacomitinib impurity 3 at 0.5, 2.5, and 12.5 mg/kg/day for 28 days per ICH Q3B guidelines. Vehicle: 0.5% methylcellulose. Endpoints include body weight, food consumption, clinical signs, serum chemistry (ALT, AST, BUN, creatinine), hematology (CBC, differential), and histopathology of skin, gastrointestinal tract (esophagus, stomach, small intestine, colon), liver, kidney, and lungs. No target‑mediated toxicities (skin rash, diarrhea) are expected. The NOAEL is expected at 12.5 mg/kg/day.
|
| ADME/Pharmacokinetics |
Pharmacokinetic profile: Dacomitinib impurity 3 (MW 402.81, log P ∼3.0‑3.5) has moderate lipophilicity. Oral absorption is expected to be moderate (40‑60%). After oral administration, Tmax ∼2‑4 h. The impurity may be metabolized via O‑demethylation (CYP2D6) and hydroxylation. The 5‑hydroxypyrrolidin-2-one group is relatively stable. The terminal half‑life is estimated to be 6‑10 h. Plasma protein binding is high (∼90%). Excretion is primarily via the biliary route as metabolites. The compound is unlikely to cross the blood‑brain barrier efficiently.
|
| Toxicity/Toxicokinetics |
Preclinical toxicology: Dacomitinib impurity 3 has moderate acute oral toxicity (estimated LD50 >500 mg/kg). In silico genotoxicity assessment (DEREK, Sarah) is required. The quinazoline core is not a structural alert for genotoxicity. An Ames test (five strains, +/-S9) and an in vitro micronucleus assay are required for ICH M7 qualification and are expected to be negative. In a 28‑day repeat‑dose toxicity study in rats, the NOAEL is expected at 2.5‑12.5 mg/kg/day. No specific target organ toxicity is anticipated due to the lack of EGFR inhibition. However, the structural similarity to dacomitinib warrants careful evaluation.
|
| Additional Infomation |
Dacomitinib impurity 3 is also known as Dacomitinib Impurity 3 and 1-(4-((3‑Chloro-4‑fluorophenyl)amino)-7‑methoxyquinazolin-6-yl)-5‑hydroxypyrrolidin-2-one. Storage: -20degC in a tightly sealed container, protected from light and moisture. Soluble in DMSO (≥10 mg/mL). Used as a reference standard for impurity profiling, stability‑indicating HPLC methods, and ANDA filings for dacomitinib API and tablets. Not for human therapeutic use.
|
| Molecular Formula |
C19H16CLFN4O3
|
|---|---|
| Molecular Weight |
402.81
|
| Exact Mass |
402.089
|
| CAS # |
2190490-31-2
|
| PubChem CID |
155896510
|
| Appearance |
White to off-white solid powder
|
| Hydrogen Bond Donor Count |
2
|
| Rotatable Bond Count |
4
|
| Heavy Atom Count |
28
|
| Complexity |
573
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
COC1=C(C=C2C(=C1)N=CN=C2NC3=CC(=C(C=C3)F)Cl)N4C(CCC4=O)O
|
| InChi Key |
YYROAISCXBYHNT-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C19H16ClFN4O3/c1-28-16-8-14-11(7-15(16)25-17(26)4-5-18(25)27)19(23-9-22-14)24-10-2-3-13(21)12(20)6-10/h2-3,6-9,17,26H,4-5H2,1H3,(H,22,23,24)
|
| Chemical Name |
1-[4-(3-chloro-4-fluoroanilino)-7-methoxyquinazolin-6-yl]-5-hydroxypyrrolidin-2-one
|
| 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: This product requires protection from light (avoid light exposure) during transportation and storage. |
| 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 | 2.4826 mL | 12.4128 mL | 24.8256 mL | |
| 5 mM | 0.4965 mL | 2.4826 mL | 4.9651 mL | |
| 10 mM | 0.2483 mL | 1.2413 mL | 2.4826 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
