Antibodies
Afatinib (BIBW2992)
- 貨號 Cat. No.
- S1011
- 規格 Size
- 5~200mg
- 品牌 Brand
- Selleckchem
Description
Afatinib (BIBW2992)
Afatinib (BIBW2992) irreversibly inhibits EGFR/HER2 including EGFR(wt), EGFR(L858R), EGFR(L858R/T790M) and HER2 with IC50 of 0.5 nM, 0.4 nM, 10 nM and 14 nM, respectively; 100-fold more active against Gefitinib-resistant L858R-T790M EGFR mutant. Phase 3.
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Distributor Tel: +886-2-2827-1197jimmy@new.abscience.com.tw
Biological Activity
| Description | Afatinib (BIBW2992) irreversibly inhibits EGFR/HER2 including EGFR(wt), EGFR(L858R), EGFR(L858R/T790M) and HER2 with IC50 of 0.5 nM, 0.4 nM, 10 nM and 14 nM, respectively; 100-fold more active against Gefitinib-resistant L858R-T790M EGFR mutant. Phase 3. | |||||
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| Targets | EGFRwt | EGFRL858R | EGFR L858R/T790M | HER2 | ||
| IC50 | 0.5 nM | 0.4 nM | 10 nM | 14 nM [1] | ||
| In vitro | BIBW2992 shows potent activity against both wild-type and mutant forms of EGFR and HER2. It is similar to Gefitinib in potency against L858R EGFR, but about 100-fold more active against the Gefitinib resistant L858R-T790M EGFR double mutant. BIBW2992 exhibits potent effects on both EGFR and HER2 phosphorylation in vivo. It compares favorably to reference compounds (such as Lapatinib et al.) in all cell types tested, such as human epidermoid carcinoma cell line A431 expressing wt EGFR, murine NIH-3T3 cells transfected with wt HER2, as well as breast cancer cell line BT-474 and gastric cancer cell line NCI-N87, which express endogenous HER2. [1] | |||||
| In vivo | Daily oral administration of BIBW2992 at 20 mg/kg for 25 days results in dramatic tumor regression with a cumulative treated/control tumor volume ratio (T/C ratio) of 2%. Reduced phosphorylation of EGFR and AKT is confirmed by immunohistochemical staining of tissue sections. Therefore, like lapatinib and neratinib, BIBW2992 is a next generation tyrosine kinase inhibitor (TKI) that inhibits human epidermal growth factor receptor 2 (Her2) and epidermal growth factor receptor (EGFR) kinases irreversibly. BIBW2992 is not only active against EGFR mutations targeted by first generation TKIs like Erlotinib or Gefitinib, but also against those insensitive to these standard therapies. [1] | |||||
| Features | ||||||
Protocol(Only for Reference)
Kinase Assay:
| In vitro kinase activity assays | The wild type tyrosine kinase domain of the human EGFR as well as that of the EGFR L858R/T790M double mutant are fused to Glutathione-S-transferase (GST), and extracted. Enzyme activity is then assayed in the presence of the inhibitor BIBW2992, serially diluted in 50% DMSO. A random polymer pEY (4:1) is used as substrate and biotinylated pEY (bio-pEY) is added as a tracer substrate. The kinase domain of HER2 is cloned using the baculovirus system and extracted similarly to that of EGFR kinase. Details of assays for EGFR, HER2, SRC, BIRK and VEGFR2 kinase activity are available in Supplementary information. |
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Cell Assay:
| Cell lines | NSCLC cells |
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| Concentrations | 0-10 μM |
| Incubation Time | 1 hour |
| Method | 1 × 104 NSCLC cells are transferred into each well of a 96-well plate and cultured overnight in serum-free media for the EGFR phosphorylation assay. After addition of BIBW2992 on the next day, the plates are incubated at 37 °C for 1 hour. EGF-stimulation is done using 100 ng/mL for 10 min at room temperature. Cells are washed with ice cold PBS, extracted with 120 μL HEPEX buffer per well and shaken at room temperature for 1 hour. In all 2 × 104 cells per well is used for the HER2 phosphorylation assay. Streptavidin precoated plates are coated with anti-EGFR-biotin at 1:100 dilution in blocking buffer and c-erb2/HER2 oncoprotein Ab-5(Clone N24)-Biotin. Cell extracts is then transferred to the antibody-coated wells and incubated at room temperature for 1 hour. Extinction is measured at 450 nm. |
Animal Study:
| Animal Models | Athymic NMRI-nu/nu female mice |
|---|---|
| Dosages | 20 mg/kg |
| Administration | Oral administration |
| Solubility | 0.5% methylcellulose/0.2% Tween 80, 30 mg/mL |
References
Clinical Trial Information( data from http://clinicaltrials.gov)
| NCT Number | Recruitment | Conditions | Sponsor /Collaborators | Start Date | Phases |
|---|---|---|---|---|---|
| NCT01836341 | Recruiting | Non-small Cell Lung Cancer | Memorial Sloan-Kettering Cancer Center|Boehringer Ingelheim Pharmaceuticals|National Comprehensive Cancer Network | 2013-04 | Phase 1 |
| NCT01814553 | Recruiting | Carcinoma, Non-Small-Cell Lung | Boehringer Ingelheim Pharmaceuticals | 2013-04 | Phase 3 |
| NCT01856478 | Recruiting | Head and Neck Neoplasms | Boehringer Ingelheim Pharmaceuticals | 2013-05 | Phase 3 |
| NCT01861223 | Not yet recruiting | NSCLC | Samsung Medical Center | 2013-05 | Phase 1|Phase 2 |
| NCT01853826 | Not yet recruiting | Carcinoma, Non-Small-Cell Lung | Boehringer Ingelheim Pharmaceuticals | 2013-06 | Phase 3 |
Chemical Information
Download Afatinib (BIBW2992) SDF
| Molecular Weight (MW) | 485.94 |
|---|---|
| Formula | C24H25ClFN5O3 |
| CAS No. | 439081-18-2 |
| Storage | 3 years -20℃Powder |
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| 6 months-80℃in DMSO | |
| Syonnyms | Tomtovok |
| Solubility (25°C) * | In vitro | DMSO | 97 mg/mL (199 mM) |
|---|---|---|---|
| Water | <1 mg/mL (<1 mM) | ||
| Ethanol | 15 mg/mL (30 mM) | ||
| In vivo | 0.5% methylcellulose/0.2% Tween 80, | 30 mg/mL | |
| * <1 mg/ml means slightly soluble or insoluble. * Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations. | |||
| Chemical Name | (S,E)-N-(4-(3-chloro-4-fluorophenylamino)-7-(tetrahydrofuran-3-yloxy)quinazolin-6-yl)-4-(dimethylamino)but-2-enamide |
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Preparing Stock Solutions
| Stock Solution (1ml DMSO) | 1mM | 10mM | 20mM | 30mM |
|---|---|---|---|---|
| Mass(mg) | 0.48594 | 4.8594 | 9.7188 | 14.5782 |
Customer Reviews (3)
 Click to enlarge | Rating | |
| Source | Int J Proteomics 2011 2011, Article ID 215496. Afatinib (BIBW2992) purchased from Selleck | |
| Method | CEER assay | |
| Cell Lines | H460/A549/RFU cell lines | |
| Concentrations | 0-10 μM | |
| Incubation Time | 4 h | |
| Results | This data showed that a potent dose-dependent inhibition of the HER1 and HER2 pathways in the HCC827 cells was observed by treatment with BIBW-2992. Other HER1 and/or HER2 pathway-activated cell lines, H1975 and H1650, were likewise had these pathway activations blocked by the treatment with BIBW-2992. |
Product Citations (10)
Non-Covalent Wild-Type-Sparing Inhibitors of EGFR T790M. [Lee HJ, et al. Cancer Discov 2013;3(2):168-81]
PubMed: 23229345
Met kinase inhibitor E7050 reverses three different mechanisms of hepatocyte growth factor-induced tyrosine kinase inhibitor resistance in EGFR mutant lung cancer. [Wang W, et al. Clin Cancer Res 2012;18(6):1663-71]
PubMed: 22317763
Akt kinase-interacting protein1, a novel therapeutic target for lung cancer with EGFR-activating and gatekeeper mutations. [Yamada T , et al. Oncogene 2013;32(37):4427-35]
PubMed: 23045273
