Size | Price | Stock | Qty |
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10mg |
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50mg |
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100mg |
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250mg |
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Other Sizes |
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Purity: ≥98%
Afatinib dimaleate (the maleate salt form of Afatinib, also know as BIBW-2992; BIBW2992) is a potent, covalent/irreversible, and orally bioavailable dual EGFR/HER2 receptor tyrosine kinase (RTK) inhibitor with potential antineoplastic activity. Afatinib is an FDA-approved anticancer medication used to treat lung cancer that is not small cell (NSCLC). In the USA, Gilotrif is the brand name under which it is sold. It is 100 times more active against the Gefitinib-resistant L858R-T790M EGFR mutant. It irreversibly binds to and inhibits EGFR/HER2, including EGFR(wt), EGFR(L858R), EGFR(L858R/T790M), and HER2. In cell-free assays, its IC50 values are 0.5 nM, 0.4 nM, 10 nM, and 14 nM, respectively.
Targets |
EGFRL858R (IC50 = 0.4 nM); EGFR (IC50 = 0.5 nM); EGFRL858R/T790M (IC50 = 10 nM); HER2 (IC50 = 14 nM); HER3
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ln Vitro |
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ln Vivo |
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Enzyme Assay |
EGFR kinase: 10 μL of inhibitor in 50% Me2SO, 20 μL of substrate solution (200 mM HEPES pH 7.4, 50 mM Mg-acetate, 2.5 mg/mL poly (EY), 5 μg/mL bio-pEY), and 20 µL enzyme preparation were included in each 100 µL enzyme reaction. The addition of 50 µL of a 100 µM ATP solution prepared in 10 mM MgCl2 initiates the enzymatic reaction. After 30 minutes of assaying at room temperature, 50 µL of stop solution (250 mM EDTA in 20 mM HEPES pH 7.4) is added to end the assay. 100 µL are added to a microtiterplate coated with streptavidin, and after 60 minutes of room temperature incubation, the plate is cleaned with 200 µL of wash solution (50 mM Tris, 0.05% Tween20). The wells are filled with a 100 µL aliquot of PY20H Anti-Ptyr:HRP, a 250 ng/mL HRPO-labeled anti-PY antibody. Following a 60-minute incubation period, the plate is three times cleaned using a 200 µL wash solution. Following that, 100µL of TMB Peroxidase Solution (A:B=1:1) is used to develop the samples. After ten minutes, the reaction is stopped. After the plate is placed in an ELISA reader, the extinction at OD450nm is calculated. The enzyme HER2-IC: The assay of enzyme activity is conducted in 50% Me2SO with or without serial inhibitor dilutions. Similar components as described for the EGFR kinase assay are included in each 100 µL reaction, along with the addition of 1000 µM Na3VO4. The addition of 50µL of a 500 µM ATP solution prepared in 10 mM magnesium acetate initiates the enzymatic reaction. The enzyme is diluted to the point where the amount of enzyme and the amount of time it takes for phosphate to be incorporated into bio-pEY are linear. The mixture of 20 mM HEPES pH 7.4, 130 mM NaCl, 0.05% Triton X-100, 1 mM DTT, and 10% glycerol is used to dilute the enzyme preparation. After 30 minutes of assaying at room temperature, 50 µL of stop solution is added to end the procedure. Src kinase assays: 10 µL of inhibitor in 50% Me2SO, 20 µL of enzyme preparation, and 20 µL of substrate solution enhanced with 1000 µM Na3VO4 were included in each 100 µL reaction. The addition of 50 µL of a 1000 µM ATP solution prepared in 10 mM Mg-acetate initiates the enzymatic reaction. Assay for BIRK kinase: 50 µL of a 2 mM ATP solution prepared in 8 mM MnCl2 and 20 mM Mg-acetate is added to 250 mM Tris pH 7.4, 10 mM DTT, 2.5 mg/mL poly(EY), and 5 mg/mL bio-pEY as the substrate solution to initiate the enzymatic reaction. HGFR kinase and VEGF2 assays: The assay is completed by adding 10 µL of 5% H3PO4 after it has been running at room temperature for 20 minutes. The precipitate is then collected using a 96 well filter mate universal harvester and trapped onto GF/B filters. The filter plate is thoroughly cleaned, dried for one hour at 50°C, sealed, and the radioactivity is measured using scintillation counting with either a TopCountTM or a Microbeta b counterTM.
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Cell Assay |
The MTT assay is used to determine cytotoxicity. The drug concentration that causes 50% of cells to die is known as the IC 50 value. The Bliss method is used to calculate the IC50 as well as the fitted sigmoidal dose response curve.
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Animal Protocol |
SCID mice harbouring ARK2 xenografts
25 mg/kg p.o. |
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References |
Molecular Formula |
C32H33CLFN5O11
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Molecular Weight |
717.18
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Exact Mass |
717.1849127
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Elemental Analysis |
C, 53.52; H, 4.63; Cl, 4.94; F, 2.65; N, 9.75; O, 24.51
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CAS # |
850140-73-7
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Related CAS # |
Afatinib;850140-72-6;Afatinib-d6 dimaleate;Afatinib oxalate;1398312-64-5
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Appearance |
White to off-white solid powder
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SMILES |
CN(C/C=C/C(=O)NC1=C(C=C2N=CN=C(C2=C1)NC3=CC(=C(C=C3)F)Cl)O[C@@H]4COCC4)C.C(=C\C(=O)O)\C(=O)O.C(=C\C(=O)O)\C(=O)O
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InChi Key |
USNRYVNRPYXCSP-JUGPPOIOSA-N
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InChi Code |
InChI=1S/C24H25ClFN5O3.2C4H4O4/c1-31(2)8-3-4-23(32)30-21-11-17-20(12-22(21)34-16-7-9-33-13-16)27-14-28-24(17)29-15-5-6-19(26)18(25)10-15;2*5-3(6)1-2-4(7)8/h3-6,10-12,14,16H,7-9,13H2,1-2H3,(H,30,32)(H,27,28,29);2*1-2H,(H,5,6)(H,7,8)/b4-3+;2*2-1-/t16-;;/m0../s1
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Chemical Name |
(Z)-but-2-enedioic acid;(E)-N-[4-(3-chloro-4-fluoroanilino)-7-[(3S)-oxolan-3-yl]oxyquinazolin-6-yl]-4-(dimethylamino)but-2-enamide
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Synonyms |
Afatinib dimaleate; BIBW 2992; BIBW-2992; BIBW2992. Afatinib free base; trade name: Gilotrif, Tomtovok and Tovok
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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 Note: Please store this product in a sealed and protected environment, 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)
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Solubility (In Vitro) |
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Solubility (In Vivo) |
Solubility in Formulation 1: 100 mg/mL (139.26 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with sonication.
Solubility in Formulation 2: 5% DMSO+30% PEG 300+ddH2O: 28 mg/mL  (Please use freshly prepared in vivo formulations for optimal results.) |
Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
1 mM | 1.3944 mL | 6.9718 mL | 13.9435 mL | |
5 mM | 0.2789 mL | 1.3944 mL | 2.7887 mL | |
10 mM | 0.1394 mL | 0.6972 mL | 1.3944 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.
NCT Number | Recruitment | interventions | Conditions | Sponsor/Collaborators | Start Date | Phases |
NCT04439136 | Active Recruiting |
Drug: Afatinib Dimaleate | Advanced Lymphoma Refractory Lymphoma |
National Cancer Institute (NCI) |
August 12, 2015 | Phase 2 |
NCT02438722 | Active Recruiting |
Drug: Afatinib Dimaleate Biological: Cetuximab |
Recurrent Non-Small Cell Lung Carcinoma Stage IV Non-Small Cell Lung Cancer |
SWOG Cancer Research Network | May 7, 2015 | Phase 2 Phase 3 |
NCT02465060 | Active Recruiting |
Drug: Afatinib Drug: Afatinib Dimaleate |
Bladder Carcinoma Glioma |
National Cancer Institute (NCI) |
August 17, 2015 | Phase 2 |
NCT03083678 | Active Recruiting |
Drug: Afatinib | Chordoma | Leiden University Medical Center | June 21, 2018 | Phase 2 |
NCT03827070 | Active Recruiting |
Drug: Afatinib Drug: Talcum powder |
Non Small Cell Lung Cancer | Center Trials & Treatment Europe | March 5, 2019 | Phase 1 |
td> |
Afatinib covalently binds to cysteine number 797 of the epidermal growth factor receptor (EGFR) via a Michael addition (IC50 = 0.5 nM).Schubert-Zsilavecz, M, Wurglics, M,Neue Arzneimittel Frühjahr 2013.(in German) td> |