| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| 10mg | |||
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| Targets |
EGFR (Epidermal Growth Factor Receptor, also known as ErbB1/HER1) Tyrosine Kinase. Gefitinib D8 is an anilinoquinazoline derivative that acts as a reversible, competitive ATP-binding inhibitor of the EGFR tyrosine kinase. By blocking the ATP-binding pocket in the intracellular domain of EGFR, it prevents receptor autophosphorylation and the subsequent activation of downstream signaling pathways (Ras-MAPK, PI3K-Akt, and JAK-STAT). This inhibits cell proliferation, angiogenesis, and induces apoptosis in EGFR-dependent cancer cells.
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| ln Vitro |
Gefitinib inhibits EGFR tyrosine kinase in cell-free assays with an IC50 of 2-37 nM in NR6wtEGFR cells (cells overexpressing wild-type EGFR). It is moderately selective for EGFR over other ErbB receptors (ErbB2/HER2). It is also an inhibitor of the ABCG2 (BCRP) efflux transporter at micromolar concentrations (IC50 ≈ 3 uM). Gefitinib induces G1 cell cycle arrest and apoptosis in NSCLC cell lines (e.g., PC9, HCC827) harboring EGFR-activating mutations (exon 19 deletions, L858R). In a BT-474 human breast carcinoma xenograft model, oral gefitinib (200 mg/kg) inhibits tumor growth significantly. In a murine model of rheumatoid arthritis, gefitinib reduces joint inflammation and damage. Gefitinib D8 is an internal standard for bioanalysis and is not tested for in vitro or in vivo activity in functional assays.
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| ln Vivo |
EGFR kinase activity is measured using a homogeneous time-resolved fluorescence (HTRF) or ELISA-based assay. Recombinant human EGFR kinase domain (0.5-5 nM) is incubated with varying concentrations of Gefitinib D8 (0.1-1000 nM) in reaction buffer (50 mM HEPES pH 7.5, 10 mM MgCl2, 2 mM MnCl2, 1 mM DTT, 0.01% BSA) containing 10-100 microM ATP for 30-60 minutes at 30degC. The reaction is initiated by adding a biotinylated peptide substrate (e.g., biotin-poly-GAT). After termination with EDTA, the reaction mixture is transferred to a streptavidin-coated plate. Phosphorylation is detected with an anti-phosphotyrosine antibody conjugated to Eu3+-cryptate (TR-FRET donor) and a fluorophore-labeled acceptor (e.g., XL665). Fluorescence is measured at 665 nm and 620 nm, and the ratio (665/620) is calculated. IC50 values are determined.
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| Enzyme Assay |
Not applicable (Gefitinib D8 is a reference standard for analytical assays, not a test article in cell-based efficacy studies). However, for the unlabeled parent: NR6wtEGFR cells (wild-type EGFR overexpressing) or A431 human epidermoid carcinoma cells (EGFR-dependent) are seeded in 96-well plates (5 × 103 cells/well) in DMEM with 1% FBS. After overnight serum starvation, cells are treated with varying concentrations of Gefitinib (unlabeled) (0.1-1000 nM) for 60 minutes, then stimulated with EGF (10-100 ng/mL) for 5-15 minutes. Cells are lysed, and EGFR phosphorylation (pEGFR, Tyr1068) is quantified by ELISA using a phospho-EGFR-specific kit (R&D Systems). The IC50 for inhibition of pEGFR is calculated. The D8-labeled version is not used in cell assays.
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| Cell Assay |
Not applicable (Gefitinib D8 is not administered in animal efficacy studies). In mouse PK studies for internal standard validation: female CD-1 mice (20-25 g, n=3 per time point) are administered gefitinib orally (10-100 mg/kg) formulated in 0.5% methylcellulose or DMSO/PEG300/Tween 80. Blood samples are collected by cardiac puncture at 0.5, 1, 2, 4, 8, and 24 h post-dose. Plasma is protein-precipitated with acetonitrile containing Gefitinib D8 as internal standard. The supernatant is injected onto an LC-MS/MS system, and gefitinib concentrations are calculated from the peak area ratio. PK parameters (Cmax, Tmax, AUC, t½) are calculated.
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| Animal Protocol |
Gefitinib D8 is a stable isotope internal standard (SIL-IS) used for LC-MS/MS quantification. The parent drug gefitinib has an oral bioavailability of approximately 50-60% in humans. It is highly protein bound (~90%), primarily to albumin and alpha1-acid glycoprotein. Gefitinib is extensively metabolized by CYP3A4, CYP2D6, and CYP2C19 to several metabolites (e.g., O-desmethyl gefitinib). The terminal elimination half-life is approximately 48 hours, allowing for once-daily dosing. The D8-labeled internal standard corrects for matrix effects and extraction losses.
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| ADME/Pharmacokinetics |
Gefitinib D8 is a research standard and not for human consumption. The parent drug gefitinib (Iressa) carries a boxed warning for hepatotoxicity and interstitial lung disease (ILD) (fatal cases reported). Common AEs (≥20%): diarrhea (87%), skin rash (acneiform, 75%), dry skin, nausea, vomiting, anorexia, and stomatitis. Less common: ocular toxicity (conjunctivitis, blepharitis), alopecia, and asthenia.
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| Toxicity/Toxicokinetics |
Gefitinib (Iressa®) was the first EGFR-TKI approved for the treatment of non-small cell lung cancer (NSCLC) (FDA approval in 2003, later restricted). Gefitinib D8 is a research internal standard used in LC-MS/MS bioanalysis for pharmacokinetic (PK) studies, bioequivalence testing, therapeutic drug monitoring (TDM), and quality control (QC) of gefitinib in pharmaceutical formulations. It is also used for the simultaneous quantification of gefitinib and its metabolites.
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| References |
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| Molecular Formula |
C₂₂H₁₆D₈CLFN₄O₃
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|---|---|
| Molecular Weight |
454.95
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| Exact Mass |
454.202
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| CAS # |
857091-32-8
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| Related CAS # |
Gefitinib;184475-35-2
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| PubChem CID |
49849312
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| Appearance |
White to off-white solid powder
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| LogP |
4.286
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
8
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| Rotatable Bond Count |
8
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| Heavy Atom Count |
31
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| Complexity |
545
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| Defined Atom Stereocenter Count |
0
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| SMILES |
[2H]C1(C(OC(C(N1CCCOC2=C(C=C3C(=C2)C(=NC=N3)NC4=CC(=C(C=C4)F)Cl)OC)([2H])[2H])([2H])[2H])([2H])[2H])[2H]
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| InChi Key |
XGALLCVXEZPNRQ-IHGLQNJRSA-N
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| InChi Code |
InChI=1S/C22H24ClFN4O3/c1-29-20-13-19-16(12-21(20)31-8-2-5-28-6-9-30-10-7-28)22(26-14-25-19)27-15-3-4-18(24)17(23)11-15/h3-4,11-14H,2,5-10H2,1H3,(H,25,26,27)/i6D2,7D2,9D2,10D2
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| Chemical Name |
N-(3-chloro-4-fluorophenyl)-7-methoxy-6-[3-(2,2,3,3,5,5,6,6-octadeuteriomorpholin-4-yl)propoxy]quinazolin-4-amine
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| Synonyms |
Gefitinib D8 Gefitinib D8
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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 | 2.1980 mL | 10.9902 mL | 21.9804 mL | |
| 5 mM | 0.4396 mL | 2.1980 mL | 4.3961 mL | |
| 10 mM | 0.2198 mL | 1.0990 mL | 2.1980 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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