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Afatinib

Manufactured by Chemietek
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Afatinib is a synthetic small-molecule tyrosine kinase inhibitor. It is designed to selectively and irreversibly bind to the epidermal growth factor receptor (EGFR) and human epidermal growth factor receptor 2 (HER2) proteins.

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Lab products found in correlation

Osimertinib, by Selleck Chemicals (2 mentions) Buparlisib, by Chemietek (1 mentions) Alpelisib, by Chemietek (1 mentions) Trametinib, by Chemietek (1 mentions) Palbociclib, by Chemietek (1 mentions) Bez235, by Chemietek (1 mentions) Rapamycin, by LC Laboratories (1 mentions) Celltiter glo luminescent cell viability assay, by Promega (1 mentions) Erlotinib, by Chemietek (1 mentions) Gefitinib, by Chemietek (1 mentions) Wz4002, by Chemietek (1 mentions) Dimedone, by Merck Group (1 mentions) Catalase, by Merck Group (1 mentions) Epidermal growth factor (egf), by BD (1 mentions) Sodium l ascorbate, by Merck Group (1 mentions) Pd168393, by Santa Cruz Biotechnology (1 mentions) Glucose oxidase, by Merck Group (1 mentions) Cuso4, by Merck Group (1 mentions) Gefitinib, by LC Laboratories (1 mentions) Erlotinib, by LC Laboratories (1 mentions)

5 protocols using afatinib

1

Anticancer Compound Screening in Mammary Cancer Cells

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Compounds for in vitro assays were obtained from Developmental Therapeutics Program at NCI (palbociclib, gedatolisib, afatinib, neratinib, buparlisib, alpelisib,), BEZ235 and trametinib from ChemieTek (Indianapolis, IN, USA), rapamycin from LC Laboratories (Woburn, MA, USA) and were dissolved in 100% DMSO. Primary mammary cancer cells were plated at 2,500 cells per well in opaque white 96-well plates. Single compound treatment was conducted at a range of concentrations between 0.1 and 50,000 nM in 0.5% DMSO. Compounds were added once to wells, 24 h after plating cells, and drug-treated wells were run in triplicate. At 72 h after drug exposure, cell viability was measured using CellTiter-Glo Luminescent Cell Viability Assay (Promega, Madison, WI, USA) as per the manufacturer’s instruction. DMSO-treated wells were considered as 100% viability for each treatment plate.
Szabova L., Gordon M.B., Lu L., Pate N., Bassel L., Iacovelli A.J., Karim B., Homan P.J., Householder D.B., Guerin T.M., Burkett S., Day A.M., Custer W, & Weaver Ohler Z. (2022). Loss of Brca1 and Trp53 in adult mouse mammary ductal epithelium results in development of hormone receptor-positive or hormone receptor-negative tumors, depending on inactivation of Rb family proteins. Breast Cancer Research : BCR, 24, 75.
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2

Tyrosine Kinase Inhibitor Evaluation

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Gefitinib, erlotinib, afatinib, and WZ4002 were purchased from Chemie Tek (Indianapolis, IN). CL-387,785 was purchased from AXXORA (San Diego, CA).
Yoshida T., Zhang G., Smith M.A., Lopez A.S., Bai Y., Li J., Fang B., Koomen J., Rawal B., Fisher K.J., Chen Y.A., Kitano M., Morita Y., Yamaguchi H., Shibata K., Okabe T., Okamoto I., Nakagawa K, & Haura E.B. (2014). Tyrosine phosphoproteomics identified both co-drivers and co-targeting strategies for T790M-related EGFR-TKI resistance in non-small cell lung cancer. Clinical cancer research : an official journal of the American Association for Cancer Research, 20(15), 4059-4074.
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3

Chemoproteomic Redox Profiling Workflow

Cited 1 time
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EGF (BD Biosciences) and H2O2 (Sigma) were prepared in ddH2O. DYn-2 (≥99% purity) was synthesized as previously described and prepared in DMSO (250 mM) (Paulsen, et al., 2012 (link)). Dimedone (250 mM, Sigma) was prepared in 70% DMSO/30% Bis-Tris (500 mM, pH 7.4). Catalase (20,000 U/ml, Sigma) and glucose oxidase (500 U/ml, Sigma) were prepared fresh in 50 mM Tris-HCl pH 7.4. Azide biotin (5 mM, Invitrogen) and TAMRA azide (5 mM, Invitrogen) were prepared in DMSO. BTTP (100 mM, Kerafast) was prepared in DMSO, and stock dilutions were prepared with ddH2O. Sodium L-ascorbate (Sigma) and CuSO4 (Sigma) were prepared fresh in ddH2O. Afatinib (10 mM, ChemieTek, ≥99% purity) and PD168393 (10 mM, Santa Cruz Biotechnology, ≥95% purity) stocks were prepared in DMSO.
Truong T.H., Ung P.M., Palde P.B., Paulsen C.E., Schlessinger A, & Carroll K.S. (2016). Molecular Basis for Redox Activation of Epidermal Growth Factor Receptor Kinase. Cell chemical biology, 23(7), 837-848.
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4

Kinase Inhibitors Comparative Analysis

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A list of the kinase inhibitors used in the experiments and the companies from which they were purchased is shown below. Afatinib (from Chemietek) was used at 0.03 nM–1 μM, Brigatinib (1–10 nM, from Shanghai Biochem), Cetuximab (10 μg/mL, from Merck), Dacomitinib (0.03 nM–1 μM, from ActiveBiochem), EAI-045 (1–10 nM, from Shanghai Biochem), Erlotinib (1–10 nM, from LC laboratories), Gefitinib (1–10 nM, from LC laboratories), Osimertinib (1–10 nM, from Selleck).
Yoshizawa T., Uchibori K., Araki M., Matsumoto S., Ma B., Kanada R., Seto Y., Oh-hara T., Koike S., Ariyasu R., Kitazono S., Ninomiya H., Takeuchi K., Yanagitani N., Takagi S., Kishi K., Fujita N., Okuno Y., Nishio M, & Katayama R. (2021). Microsecond-timescale MD simulation of EGFR minor mutation predicts the structural flexibility of EGFR kinase core that reflects EGFR inhibitor sensitivity. NPJ Precision Oncology, 5, 32.
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5

Inhibitor Compound Acquisition and Preparation

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Lorlatinib, crizotinib, gilteritinib, and brigatinib were purchased from Shanghai Biochempartner Co., Ltd. Alectinib was purchased from ActiveBiochem. Taletrectinib and entrectinib were synthesized at DaiichiSankyo Co., Ltd. Afatinib was obtained from ChemieTek. entrectinib was purchased from MedChemExpress. PP242 was bought from AdooQ Bioscience. Osimertinib was purchased from Selleck. Panitumumab was procured from Takeda Pharm. The human recombinants EGF, HB-EGF, and TGF-α were purchased from PeproTech. Brigatinib was dissolved in ethanol, and the other inhibitors were dissolved in dimethyl sulfoxide (DMSO) for the cell culture experiments.
Kondo N., Utsumi T., Shimizu Y., Takemoto A., Oh-hara T., Uchibori K., Subat-Motoshi S., Ninomiya H., Takeuchi K., Nishio M., Miyazaki Y, & Katayama R. (2023). MIG6 loss confers resistance to ALK/ROS1 inhibitors in NSCLC through EGFR activation by low-dose EGF. JCI Insight, 8(24), e173688.
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