Afatinib (BIBW2992) Dimaleate

Name: Afatinib (BIBW2992) Dimaleate
Brand: Selleck Chemicals
SKU: S7810
Rating: 5 (121 reviews)

For research use only.

Catalog No.S7810

121 publications

Afatinib (BIBW2992) Dimaleate Chemical Structure

CAS No. 850140-73-7

Afatinib (BIBW2992) Dimaleate 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. Afatinib (BIBW2992) Dimaleate induces autophagy.

Selleck's Afatinib (BIBW2992) Dimaleate has been cited by 121 publications

Cancer Cell, 2021, S1535-6108(21)00284-1

PubMed: 34171264 ( click the link to review the publication )

Nat Cell Biol, 2021, 23(4):377-390

PubMed: 33795873 ( click the link to review the publication )

Cancer Cell, 2020, 37(2):183-199

PubMed: 31978326 ( click the link to review the publication )

Cancer Discov, 2017, 7(6):575-585

PubMed: 28274957 ( click the link to review the publication )

Cancer Discov, 2013, 3(2):168-81

PubMed: 23229345 ( click the link to review the publication )

Theranostics, 2021, 11(6):2594-2611

PubMed: 33456562 ( click the link to review the publication )

Mol Cancer, 2021, 20(1):17

PubMed: 33461557 ( click the link to review the publication )

Cancers (Basel), 2021, 13(14)3635

PubMed: 34298852 ( click the link to review the publication )

Cell Death Dis, 2021, 12(8):728

PubMed: 34294686 ( click the link to review the publication )

Pharmacol Res, 2021, 170:105701

PubMed: 34087353 ( click the link to review the publication )

Front Oncol, 2021, 11:665045

PubMed: 34168988 ( click the link to review the publication )

Biochem Pharmacol, 2021, 190:114642

PubMed: 34077739 ( click the link to review the publication )

Int J Mol Sci, 2021, 22(8)4005

PubMed: 33924522 ( click the link to review the publication )

Drug Metab Dispos, 2021, 49(1):53-61

PubMed: 33148688 ( click the link to review the publication )

Cancer Res Treat, 2021, 10.4143/crt.2021.385

PubMed: 33940786 ( click the link to review the publication )

Transl Oncol, 2021, 14(8):101110

PubMed: 33951601 ( click the link to review the publication )

Onco Targets Ther, 2021, 14:3813-3820

PubMed: 34188489 ( click the link to review the publication )

Thorac Cancer, 2021, 10.1111/1759-7714.13931

PubMed: 33710795 ( click the link to review the publication )

Jpn J Clin Oncol, 2021, hyab048

PubMed: 33829270 ( click the link to review the publication )

Oncol Lett, 2021, 21(5):418

PubMed: 33841579 ( click the link to review the publication )

Hum Cell, 2021, 10.1007/s13577-021-00579-z

PubMed: 34304386 ( click the link to review the publication )

Lung Cancer, 2021, 154:84-91

PubMed: 33631449 ( click the link to review the publication )

Nat Nanotechnol, 2021, 10.1038/s41565-021-00872-w

PubMed: 33686255 ( click the link to review the publication )

PLoS Pathog, 2021, 17(2):e1009303

PubMed: 33529218 ( click the link to review the publication )
Toxicol Lett, 2021, S0378-4274(21)00043-6

PubMed: 33571620 ( click the link to review the publication )

Clin Cancer Res, 2020, clincanres.3555.2020

PubMed: 33355298 ( click the link to review the publication )

Genome Med, 2020, 18;12(1):17

PubMed: 32070411 ( click the link to review the publication )

Oncogene, 2020, 10.1038/s41388-020-01564-w

PubMed: 33288882 ( click the link to review the publication )

Oncogene, 2020, 39(20):3997-4013

PubMed: 32242147 ( click the link to review the publication )

J Invest Dermatol, 2020, 3 pii: S0022-202X(20)31407-X

PubMed: 32376279 ( click the link to review the publication )

Cancers (Basel), 2020, 13;12(2) pii: E436

PubMed: 32069833 ( click the link to review the publication )

Cancers (Basel), 2020, 12(12)E3869

PubMed: 33371412 ( click the link to review the publication )

Cell Death Dis, 2020, 11(10):882

PubMed: 33082316 ( click the link to review the publication )

Signal Transduct Target Ther, 2020, 5(1):214

PubMed: 33033232 ( click the link to review the publication )

Oral Oncol, 2020, 112:105074

PubMed: 33142224 ( click the link to review the publication )

J Virol, 2020, JVI.02001-20

PubMed: 33361428 ( click the link to review the publication )

Antimicrob Agents Chemother, 2020, 1;AAC.00341-20

PubMed: 32482675 ( click the link to review the publication )

Sci Rep, 2020, 10(1):17768

PubMed: 33082482 ( click the link to review the publication )

J Biol Chem, 2020, jbc.RA120.015891

PubMed: 33087447 ( click the link to review the publication )

Am J Transl Res, 2020, 15;12(3):847-856 eCollection 2020

PubMed: 32269717 ( click the link to review the publication )

Cancer Res Treat, 2020, 10.4143/crt.2020.585

PubMed: 33070556 ( click the link to review the publication )

Toxicol In Vitro, 2020, 104928

PubMed: 32622998 ( click the link to review the publication )

Oncol Rep, 2020, 44(6):2581-2594

PubMed: 33125153 ( click the link to review the publication )

PLoS One, 2020, 15(1):e0227261

PubMed: 31914141 ( click the link to review the publication )

Hum Cell, 2020, 10.1007/s13577-020-00420-z

PubMed: 32870449 ( click the link to review the publication )

Anticancer Res, 2020, 40(4):1867-1874

PubMed: 32234874 ( click the link to review the publication )

Korean J Parasitol, 2020, 58(3):249-255

PubMed: 32615738 ( click the link to review the publication )

Neurooncol Adv, 2020, Dec;2(1):vdaa020

PubMed: 32226941 ( click the link to review the publication )
Sci Rep, 2020, 10(1):12982

PubMed: 32737414 ( click the link to review the publication )

BMC Pharmacol Toxicol, 2020, 21(1):61

PubMed: 32795383 ( click the link to review the publication )

NPJ Breast Cancer, 2020, 6:30

PubMed: 32715085 ( click the link to review the publication )

Physiol Rep, 2020, 8(13):e14490

PubMed: 32652816 ( click the link to review the publication )

J Cell Physiol, 2020, 10.1002/jcp.30172

PubMed: 33230821 ( click the link to review the publication )

Rapid Commun Mass Spectrom, 2020, 10.1002/rcm.8955

PubMed: 32990383 ( click the link to review the publication )

Cancer Cell Int, 2020, 20:376

PubMed: 32774169 ( click the link to review the publication )

Circulation, 2019, 139(22):2570-2584

PubMed: 30922063 ( click the link to review the publication )

Oncoimmunology, 2019,

PubMed: 30713780 ( click the link to review the publication )

Sci Rep, 2019,

PubMed: 31882684 ( click the link to review the publication )

EMBO Mol Med, 2018, 10(9):e8746

PubMed: 30021798 ( click the link to review the publication )

Clin Cancer Res, 2018, 24(5):1227-1239

PubMed: 29229632 ( click the link to review the publication )

Clin Cancer Res, 2018, 15;24(6):1436-1447

PubMed: 29127119 ( click the link to review the publication )

Cell Death Dis, 2018,

PubMed: 29880898 ( click the link to review the publication )

Mol Cancer Ther, 2018, 17(8):1648-1658

PubMed: 29748209 ( click the link to review the publication )

J Thorac Oncol, 2017, 12(5):884-889

PubMed: 28088511 ( click the link to review the publication )

Theranostics, 2017, 7(4):974-986

PubMed: 28382169 ( click the link to review the publication )

Oncogene, 2017, 36(47):6581-6591

PubMed: 28783173 ( click the link to review the publication )

Arch Toxicol, 2017, 91(8):2921-2938

PubMed: 28032146 ( click the link to review the publication )

Mol Cancer Ther, 2017,

PubMed: 27913578 ( click the link to review the publication )

BMC Cancer, 2017,

PubMed: 28806950 ( click the link to review the publication )

Oncotarget, 2017, 8(3):4277-4288

PubMed: 28032592 ( click the link to review the publication )

Oncotarget, 2017, 8(14):22414-22432

PubMed: 26461472 ( click the link to review the publication )

Oncotarget, 2017,

PubMed: 29050315 ( click the link to review the publication )
Oncotarget, 2017,

PubMed: 28423495 ( click the link to review the publication )

Mol Cancer Ther, 2016,

PubMed: 27207775 ( click the link to review the publication )

Mol Cancer Ther, 2016,

PubMed: 27678331 ( click the link to review the publication )

Cancer Sci, 2016, 107(8):1134-40

PubMed: 27240419 ( click the link to review the publication )

Cancer Immunol Res, 2016,

PubMed: 27799141 ( click the link to review the publication )

Cancer Immunol Res, 2016,

PubMed: 27680026 ( click the link to review the publication )

Oncotarget, 2016,

PubMed: 26623721 ( click the link to review the publication )

Gut, 2015, 10.1136/gutjnl-2014-309026

PubMed: ( click the link to review the publication )

Nat Commun, 2015, 6:8481

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Clin Cancer Res, 2015, 10.1158/1078-0432.CCR-14-2789

PubMed: 25948633 ( click the link to review the publication )

Clin Cancer Res, 2015, 10.1158/1078-0432.CCR-15-0560

PubMed: 25964297 ( click the link to review the publication )

J Cell Biol, 2015, 211(2):391-405

PubMed: 26483553 ( click the link to review the publication )

Cell Rep, 2015, 11(1):98-110

PubMed: 25843712 ( click the link to review the publication )

Int J Cancer, 2015, 136(11):2717-29

PubMed: ( click the link to review the publication )

J Med Chem, 2015, 58(16):6589-606

PubMed: 26258521 ( click the link to review the publication )

Cancer Discov, 2015,

PubMed: 26243863 ( click the link to review the publication )

Mol Cancer Ther, 2015, 10.1158/1535-7163.MCT-13-0951

PubMed: 25589492 ( click the link to review the publication )

Antimicrob Agents Chemother, 2015, 59(2):1088-99

PubMed: 25487801 ( click the link to review the publication )

Antimicrob Agents Chemother, 2015, 59(2):1088-99

PubMed: ( click the link to review the publication )

Drug Metab Dispos, 2015, 43(3):375-84

PubMed: ( click the link to review the publication )

Drug Metab Dispos, 2015, 43(3):375-84

PubMed: 25504185 ( click the link to review the publication )

Cell Tissue Res, 2015, 359(3):799-816

PubMed: ( click the link to review the publication )

Oncol Rep, 2015,

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Arch Immunol Ther Exp, 2015, 63(3):207-14

PubMed: 25678473 ( click the link to review the publication )
PLoS ONE, 2015,

PubMed: 25969993 ( click the link to review the publication )

Anticancer Res, 2015, 35(4):2005-8

PubMed: 25862853 ( click the link to review the publication )

Oncotarget, 2015,

PubMed: 25686822 ( click the link to review the publication )

Oncotarget, 2015,

PubMed: 25965827 ( click the link to review the publication )

Oncotarget, 2015,

PubMed: 25537503 ( click the link to review the publication )

Nat Commun, 2014, 5:5232

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J Natl Cancer Inst, 2014, 106(9)

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Clin Cancer Res, 2014, 20(21):5423-34

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Anal Chem, 2014, 86(21):10568-75

PubMed: 25263480 ( click the link to review the publication )

Drug Metab Dispos, 2014, 42(11):1851-7

PubMed: 25165131 ( click the link to review the publication )

Histochem Cell Biol, 2014, 142(4):361-71

PubMed: 24824474 ( click the link to review the publication )

Target Oncol, 2014, 10.1007/s11523-014-0344-7

PubMed: 25341405 ( click the link to review the publication )

Oncotarget, 2014, 5(24):12877-90

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Assay Drug Dev Technol, 2014, 12(9-10):514-26

PubMed: 25506801 ( click the link to review the publication )

Oncotarget, 2014, 5(15):5908-19

PubMed: 25115383 ( click the link to review the publication )

Genes Cancer, 2014, 5(7-8):261-72

PubMed: 25221644 ( click the link to review the publication )

Eur J Cancer, 2013, 49(6):1458-66

PubMed: 23153706 ( click the link to review the publication )

Oncogene, 2013, 32(37):4427-35

PubMed: 23045273 ( click the link to review the publication )

Cancer Sci, 2013, 104(12):1618-25

PubMed: 24112719 ( click the link to review the publication )

Evid Based Complement Alternat Med, 2013, 2013:243859

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J Thorac Oncol, 2012, 7(2):272-80

PubMed: 22089117 ( click the link to review the publication )

Clin Cancer Res, 2012, 18(6):1663-71

PubMed: 22317763 ( click the link to review the publication )

PLoS One, 2012, 7(7):e41017

PubMed: 22815900 ( click the link to review the publication )

Oncol Res, 2011, 19(10-11):471-8

PubMed: 22715590 ( click the link to review the publication )
Int J Proteomics, 2011, 2011:215496

PubMed: 22091388 ( click the link to review the publication )

Purity & Quality Control

Choose Selective EGFR Inhibitors

Biological Activity

Description

Targets

EGFR (L858R) ^[1] (Cell-free assay)	EGFR (wt) ^[1] (Cell-free assay)	EGFR (L858R/T790M) ^[1] (Cell-free assay)	HER2 ^[1] (Cell-free assay)
0.4 nM	0.5 nM	10 nM	14 nM

In vitro

BIBW2992 is more effective than erlotinib, geﬁtinib, or lapatinib in inhibiting survival of lung cancer cell lines harboring wild-type (H1666) or L858R/T790M (NCI-H1975) EGFR. BIBW2992 is similarly effective against NSCLC lines expressing HER2 776insV (NCI-H1781) or EGFR E746_A750del (HCC827), but shows no activity toward A549 cells, which express wild-type EGFR and HER2.^[1] Afatinib enhances cytotoxicity of topotecan and mitoxantrone to SP cells, and increases the apoptosis induced by topotecan and mitoxantrone in SP cells.^[2]

Cell Data

Cell Lines	Assay Type	Incubation Time	Activity Description	PMID
HCC827 cell	Ml7TVJJwdGmoZYLheIlwdiCjc4PhfS=>		M2rnRWFvfGmycn;sbYZmemG2aY\lJIFkfGm4aYT5JIFo[Wmwc4SgbJVu[W5iSFPDPFI4KGOnbHzzJIFnfGW{IEm2JIhzeyCkeTDNWHMh[XO\|YYmsJGlEPTB;ODDuUS=>	NWj3NY9XOjB3NUCyNVI>
NCI-H1563	NF3xOFdIem:5dHigbY5pcWKrdHnvckBie3OjeR?=		NIjYPJdKdmirYnn0bY9vKG:oIHj1cYFvKE6FST3INVU3OyClZXzsJIdzd3e2aDDpckBiKGOnbHygeoli[mmuaYT5JIF{e2G7LjDJR\|UxRTlwN{[wN\|Eh\|ryP	MVrTRW5ITVJ?
NCI-H1573	MVzHdo94fGhiaX7obYJqfGmxbjDhd5NigQ>?		NGfhUYhKdmirYnn0bY9vKG:oIHj1cYFvKE6FST3INVU4OyClZXzsJIdzd3e2aDDpckBiKGOnbHygeoli[mmuaYT5JIF{e2G7LjDJR\|UxRTBwN{O2N{DPxE1?	MVXTRW5ITVJ?
NCI-H1581	NVnqbFV2T3Kxd4ToJIlvcGmkaYTpc44h[XO\|YYm=		M{XrbGlvcGmkaYTpc44hd2ZiaIXtZY4hVkOLLVixOVgyKGOnbHyg[5Jwf3SqIHnuJIEh[2WubDD2bYFjcWyrdImgZZN{[XlwIFnDOVA:OjlwM{G0OUDPxE1?	MnjNV2FPT0WU
NCI-H1648	MnzlS5Jwf3SqIHnubIljcXSrb36gZZN{[Xl?		MnfPTY5pcWKrdHnvckBw\iCqdX3hckBPS0lvSEG2OFgh[2WubDDndo94fGhiaX6gZUBk\WyuII\pZYJqdGm2eTDhd5NigS5iSVO1NF0xNjR2MUiyJO69VQ>?	M1rH[HNCVkeHUh?=
NCI-H1793	MkLwS5Jwf3SqIHnubIljcXSrb36gZZN{[Xl?		NGHpTY1KdmirYnn0bY9vKG:oIHj1cYFvKE6FST3INVc6OyClZXzsJIdzd3e2aDDpckBiKGOnbHygeoli[mmuaYT5JIF{e2G7LjDJR\|UxRTF2LkKwPFch\|ryP	MUfTRW5ITVJ?
NCI-H1838	M1rrZWdzd3e2aDDpcohq[mm2aX;uJIF{e2G7		NEC4eWZKdmirYnn0bY9vKG:oIHj1cYFvKE6FST3INVg{QCClZXzsJIdzd3e2aDDpckBiKGOnbHygeoli[mmuaYT5JIF{e2G7LjDJR\|UxRTBwNkCyPVkh\|ryP	NIOyNYFUSU6JRWK=
NCI-H1770	NYLRcVFkT3Kxd4ToJIlvcGmkaYTpc44h[XO\|YYm=		Mnj4TY5pcWKrdHnvckBw\iCqdX3hckBPS0lvSEG3O\|Ah[2WubDDndo94fGhiaX6gZUBk\WyuII\pZYJqdGm2eTDhd5NigS5iSVO1NF0xNjFyMkig{txO	NETQcINUSU6JRWK=
NCI-H1651	M{\Zemdzd3e2aDDpcohq[mm2aX;uJIF{e2G7		MUHJcohq[mm2aX;uJI9nKGi3bXHuJG5EUS2KMU[1NUBk\WyuIHfyc5d1cCCrbjDhJINmdGxidnnhZoltcXS7IHHzd4F6NiCLQ{WwQVI2NjV7NzFOwG0>	NX;FVlRCW0GQR1XS
NCI-H1975	MoT1S5Jwf3SqIHnubIljcXSrb36gZZN{[Xl?		MkLMTY5pcWKrdHnvckBw\iCqdX3hckBPS0lvSEG5O\|Uh[2WubDDndo94fGhiaX6gZUBk\WyuII\pZYJqdGm2eTDhd5NigS5iSVO1NF0xNjZzN{K4JO69VQ>?	M{HGb3NCVkeHUh?=
NCI-H1666	MYjHdo94fGhiaX7obYJqfGmxbjDhd5NigQ>?		M3e0dGlvcGmkaYTpc44hd2ZiaIXtZY4hVkOLLVixOlY3KGOnbHyg[5Jwf3SqIHnuJIEh[2WubDD2bYFjcWyrdImgZZN{[XlwIFnDOVA:OS56MEm0PEDPxE1?	MYrTRW5ITVJ?
NCI-H2009	NGPWbYZIem:5dHigbY5pcWKrdHnvckBie3OjeR?=		Ml;PTY5pcWKrdHnvckBw\iCqdX3hckBPS0lvSEKwNFkh[2WubDDndo94fGhiaX6gZUBk\WyuII\pZYJqdGm2eTDhd5NigS5iSVO1NF0{NjZzMkG2JO69VQ>?	M3rXUnNCVkeHUh?=
NCI-H2126	NVrEdXdJT3Kxd4ToJIlvcGmkaYTpc44h[XO\|YYm=		Mln3TY5pcWKrdHnvckBw\iCqdX3hckBPS0lvSEKxNlYh[2WubDDndo94fGhiaX6gZUBk\WyuII\pZYJqdGm2eTDhd5NigS5iSVO1NF0xNjVzOUK2JO69VQ>?	M2jrRXNCVkeHUh?=
NCI-H1693	MnvaS5Jwf3SqIHnubIljcXSrb36gZZN{[Xl?		NU\GXXBxUW6qaXLpeIlwdiCxZjDoeY1idiCQQ1mtTFE3QTNiY3XscEBoem:5dHigbY4h[SClZXzsJJZq[WKrbHn0fUBie3OjeT6gTWM2OD1\|Lki0NVgyKM7:TR?=	MlnuV2FPT0WU
NCI-H2087	M3nhfGdzd3e2aDDpcohq[mm2aX;uJIF{e2G7		MWXJcohq[mm2aX;uJI9nKGi3bXHuJG5EUS2KMkC4O{Bk\WyuIHfyc5d1cCCrbjDhJINmdGxidnnhZoltcXS7IHHzd4F6NiCLQ{WwQVQ6Njl4OEKg{txO	M3vMXXNCVkeHUh?=
NCI-H2029	M3vvW2dzd3e2aDDpcohq[mm2aX;uJIF{e2G7		MYjJcohq[mm2aX;uJI9nKGi3bXHuJG5EUS2KMkCyPUBk\WyuIHfyc5d1cCCrbjDhJINmdGxidnnhZoltcXS7IHHzd4F6NiCLQ{WwQVQ{NjV3OEGg{txO	M2XBSHNCVkeHUh?=
NCI-H2170	NYnzTYFxT3Kxd4ToJIlvcGmkaYTpc44h[XO\|YYm=		NWrWc3RxUW6qaXLpeIlwdiCxZjDoeY1idiCQQ1mtTFIyPzBiY3XscEBoem:5dHigbY4h[SClZXzsJJZq[WKrbHn0fUBie3OjeT6gTWM2OD1yLkCzO\|I5KM7:TR?=	NGjrc3FUSU6JRWK=
NCI-H226	NGHSN2hIem:5dHigbY5pcWKrdHnvckBie3OjeR?=		MXXJcohq[mm2aX;uJI9nKGi3bXHuJG5EUS2KMkK2JINmdGxiZ4Lve5RpKGmwIHGgZ4VtdCC4aXHibYxqfHliYYPzZZkvKEmFNUC9OFkvPzd6NTFOwG0>	MkLtV2FPT0WU
NCI-H2030	M3fvZmdzd3e2aDDpcohq[mm2aX;uJIF{e2G7		NFzZd5VKdmirYnn0bY9vKG:oIHj1cYFvKE6FST3INlA{OCClZXzsJIdzd3e2aDDpckBiKGOnbHygeoli[mmuaYT5JIF{e2G7LjDJR\|UxRTF2Lk[0OFgh\|ryP	NVHDOolzW0GQR1XS
NCI-H2052	NW\iV5F3T3Kxd4ToJIlvcGmkaYTpc44h[XO\|YYm=		NX[zT2xGUW6qaXLpeIlwdiCxZjDoeY1idiCQQ1mtTFIxPTJiY3XscEBoem:5dHigbY4h[SClZXzsJJZq[WKrbHn0fUBie3OjeT6gTWM2OD1{MT64PVY{KM7:TR?=	NGTlc\|FUSU6JRWK=
NCI-H292	NUPscIxvT3Kxd4ToJIlvcGmkaYTpc44h[XO\|YYm=		MYDJcohq[mm2aX;uJI9nKGi3bXHuJG5EUS2KMkmyJINmdGxiZ4Lve5RpKGmwIHGgZ4VtdCC4aXHibYxqfHliYYPzZZkvKEmFNUC9NE4xPjB7IN88US=>	NV7DWmhHW0GQR1XS
NCI-H441	M3HiUmdzd3e2aDDpcohq[mm2aX;uJIF{e2G7		NHfUNGRKdmirYnn0bY9vKG:oIHj1cYFvKE6FST3IOFQyKGOnbHyg[5Jwf3SqIHnuJIEh[2WubDD2bYFjcWyrdImgZZN{[XlwIFnDOVA:Oy53OUi0NUDPxE1?	MkDpV2FPT0WU
NCI-H2291	NXrt[ZBGT3Kxd4ToJIlvcGmkaYTpc44h[XO\|YYm=		MXTJcohq[mm2aX;uJI9nKGi3bXHuJG5EUS2KMkK5NUBk\WyuIHfyc5d1cCCrbjDhJINmdGxidnnhZoltcXS7IHHzd4F6NiCLQ{WwQVUvQTh7NEOg{txO	NWr6XG1vW0GQR1XS
NCI-H358	MlLoS5Jwf3SqIHnubIljcXSrb36gZZN{[Xl?		M4fXb2lvcGmkaYTpc44hd2ZiaIXtZY4hVkOLLVizOVgh[2WubDDndo94fGhiaX6gZUBk\WyuII\pZYJqdGm2eTDhd5NigS5iSVO1NF0xNjV\|MkGg{txO	NIDXV2ZUSU6JRWK=
NCI-H460	NXnDcFFHT3Kxd4ToJIlvcGmkaYTpc44h[XO\|YYm=		MUDJcohq[mm2aX;uJI9nKGi3bXHuJG5EUS2KNE[wJINmdGxiZ4Lve5RpKGmwIHGgZ4VtdCC4aXHibYxqfHliYYPzZZkvKEmFNUC9OFYvPTJ\|IN88US=>	NXv0OG5LW0GQR1XS
NCI-H661	Mn;xS5Jwf3SqIHnubIljcXSrb36gZZN{[Xl?		M2KwW2lvcGmkaYTpc44hd2ZiaIXtZY4hVkOLLVi2OlEh[2WubDDndo94fGhiaX6gZUBk\WyuII\pZYJqdGm2eTDhd5NigS5iSVO1NF0yPi57Nkm0JO69VQ>?	NHvMfWJUSU6JRWK=
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NCI-N87	Mki2S5Jwf3SqIHnubIljcXSrb36gZZN{[Xl?		MYnJcohq[mm2aX;uJI9nKGi3bXHuJG5EUS2QOEegZ4VtdCCpcn;3eIghcW5iYTDj[YxtKH[rYXLpcIl1gSCjc4PhfU4hUUN3ME2wMlAzPzd\|IN88US=>	NYrBZpVNW0GQR1XS
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NCI-H2122	NIHONXBIem:5dHigbY5pcWKrdHnvckBie3OjeR?=		MV;Jcohq[mm2aX;uJI9nKGi3bXHuJG5EUS2KMkGyNkBk\WyuIHfyc5d1cCCrbjDhJINmdGxidnnhZoltcXS7IHHzd4F6NiCLQ{WwQVM1NjN2N{Gg{txO	NHnB[m9USU6JRWK=
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OVCAR-3	M4G5OWdzd3e2aDDpcohq[mm2aX;uJIF{e2G7		M1;JbWlvcGmkaYTpc44hd2ZiaIXtZY4hV1[FQWKtN{Bk\WyuIHfyc5d1cCCrbjDhJINmdGxidnnhZoltcXS7IHHzd4F6NiCLQ{WwQVIvPjdzNkeg{txO	MkHjV2FPT0WU
OVCAR-4	MmD2S5Jwf3SqIHnubIljcXSrb36gZZN{[Xl?		MnLOTY5pcWKrdHnvckBw\iCqdX3hckBQXkODUj20JINmdGxiZ4Lve5RpKGmwIHGgZ4VtdCC4aXHibYxqfHliYYPzZZkvKEmFNUC9NVAvQDh\|NTFOwG0>	MYnTRW5ITVJ?
OVCAR-5	NV3YN2lMT3Kxd4ToJIlvcGmkaYTpc44h[XO\|YYm=		M2jOVGlvcGmkaYTpc44hd2ZiaIXtZY4hV1[FQWKtOUBk\WyuIHfyc5d1cCCrbjDhJINmdGxidnnhZoltcXS7IHHzd4F6NiCLQ{WwQVE{NjF3OUOg{txO	NWPD[nNqW0GQR1XS
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PC-14	MnPxS5Jwf3SqIHnubIljcXSrb36gZZN{[Xl?		MWDJcohq[mm2aX;uJI9nKGi3bXHuJHBENTF2IHPlcIwh\3Kxd4ToJIlvKGFiY3XscEB3cWGkaXzpeJkh[XO\|YYmuJGlEPTB;MD6wOVY3QCEQvF2=	NWr4cZp3W0GQR1XS
PANC-03-27	MXfHdo94fGhiaX7obYJqfGmxbjDhd5NigQ>?		NWOzdoc1UW6qaXLpeIlwdiCxZjDoeY1idiCSQV7DMVA{NTJ5IHPlcIwh\3Kxd4ToJIlvKGFiY3XscEB3cWGkaXzpeJkh[XO\|YYmuJGlEPTB;Mj6xOlY{OSEQvF2=	NEHy[JNUSU6JRWK=
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NCI-H727	MmPSS5Jwf3SqIHnubIljcXSrb36gZZN{[Xl?		NUiyVVM6UW6qaXLpeIlwdiCxZjDoeY1idiCQQ1mtTFczPyClZXzsJIdzd3e2aDDpckBiKGOnbHygeoli[mmuaYT5JIF{e2G7LjDJR\|UxRThwMUC0NkDPxE1?	MkXGV2FPT0WU
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RERF-LC-MS	Moe5S5Jwf3SqIHnubIljcXSrb36gZZN{[Xl?		MkD3TY5pcWKrdHnvckBw\iCqdX3hckBTTVKILVzDMW1UKGOnbHyg[5Jwf3SqIHnuJIEh[2WubDD2bYFjcWyrdImgZZN{[XlwIFnDOVA:OTdwNUWwPEDPxE1?	NH;hOIJUSU6JRWK=
RCC10RGB	MnziS5Jwf3SqIHnubIljcXSrb36gZZN{[Xl?		NUXWdopJUW6qaXLpeIlwdiCxZjDoeY1idiCUQ1OxNHJISiClZXzsJIdzd3e2aDDpckBiKGOnbHygeoli[mmuaYT5JIF{e2G7LjDJR\|UxRTF6LkC1OVkh\|ryP	MnnQV2FPT0WU
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NCI-H1355	NGOweXNIem:5dHigbY5pcWKrdHnvckBie3OjeR?=		NFu2SppKdmirYnn0bY9vKG:oIHj1cYFvKE6FST3INVM2PSClZXzsJIdzd3e2aDDpckBiKGOnbHygeoli[mmuaYT5JIF{e2G7LjDJR\|UxRTN2LkO4PFkh\|ryP	NWTLPYs1W0GQR1XS
NCI-H1792	M{PsPWdzd3e2aDDpcohq[mm2aX;uJIF{e2G7		MnmxTY5pcWKrdHnvckBw\iCqdX3hckBPS0lvSEG3PVIh[2WubDDndo94fGhiaX6gZUBk\WyuII\pZYJqdGm2eTDhd5NigS5iSVO1NF02OC5{N{S0JO69VQ>?	MVnTRW5ITVJ?
NCI-H2347	MmP0S5Jwf3SqIHnubIljcXSrb36gZZN{[Xl?		NXn4SZlpUW6qaXLpeIlwdiCxZjDoeY1idiCQQ1mtTFI{PDdiY3XscEBoem:5dHigbY4h[SClZXzsJJZq[WKrbHn0fUBie3OjeT6gTWM2OD13OD6zOlQ5KM7:TR?=	MWTTRW5ITVJ?
NCI-H2452	NXfkUJF7T3Kxd4ToJIlvcGmkaYTpc44h[XO\|YYm=		MWXJcohq[mm2aX;uJI9nKGi3bXHuJG5EUS2KMkS1NkBk\WyuIHfyc5d1cCCrbjDhJINmdGxidnnhZoltcXS7IHHzd4F6NiCLQ{WwQVU3NjV6MUmg{txO	NYDtXoFzW0GQR1XS
PSN1	NITS[VNIem:5dHigbY5pcWKrdHnvckBie3OjeR?=		Mlm2TY5pcWKrdHnvckBw\iCqdX3hckBRW05zIHPlcIwh\3Kxd4ToJIlvKGFiY3XscEB3cWGkaXzpeJkh[XO\|YYmuJGlEPTB;NUCuPVM6PCEQvF2=	MX;TRW5ITVJ?
RH-1	MX3Hdo94fGhiaX7obYJqfGmxbjDhd5NigQ>?		M3\EemlvcGmkaYTpc44hd2ZiaIXtZY4hWkhvMTDj[YxtKGe{b4f0bEBqdiCjIHPlcIwhfmmjYnnsbZR6KGG\|c3H5MkBKSzVyPU[2MlMzPzdizszN	NU[5UpNjW0GQR1XS
SK-LMS-1	NYLqT3hLT3Kxd4ToJIlvcGmkaYTpc44h[XO\|YYm=		NWnaZlFuUW6qaXLpeIlwdiCxZjDoeY1idiCVSz3MUXMuOSClZXzsJIdzd3e2aDDpckBiKGOnbHygeoli[mmuaYT5JIF{e2G7LjDJR\|UxRTVzLkm4NVUh\|ryP	MULTRW5ITVJ?
SK-MEL-30	MVjHdo94fGhiaX7obYJqfGmxbjDhd5NigQ>?		NGruUJJKdmirYnn0bY9vKG:oIHj1cYFvKFONLV3FUE0{OCClZXzsJIdzd3e2aDDpckBiKGOnbHygeoli[mmuaYT5JIF{e2G7LjDJR\|UxRTF\|Mz60PFEh\|ryP	NVzLV5RSW0GQR1XS
SNB75	MnnGS5Jwf3SqIHnubIljcXSrb36gZZN{[Xl?		M1jmNWlvcGmkaYTpc44hd2ZiaIXtZY4hW06EN{WgZ4VtdCCpcn;3eIghcW5iYTDj[YxtKH[rYXLpcIl1gSCjc4PhfU4hUUN3ME21NE41OTRizszN	MnvFV2FPT0WU
SNU-423	MVLHdo94fGhiaX7obYJqfGmxbjDhd5NigQ>?		MUHJcohq[mm2aX;uJI9nKGi3bXHuJHNPXS12MkOgZ4VtdCCpcn;3eIghcW5iYTDj[YxtKH[rYXLpcIl1gSCjc4PhfU4hUUN3ME23NE43ODF5IN88US=>	MXfTRW5ITVJ?
SW1088	M4G4XGdzd3e2aDDpcohq[mm2aX;uJIF{e2G7		NYHVV\|dLUW6qaXLpeIlwdiCxZjDoeY1idiCVV{GwPFgh[2WubDDndo94fGhiaX6gZUBk\WyuII\pZYJqdGm2eTDhd5NigS5iSVO1NF02OC5yMkCxJO69VQ>?	NFPiZlZUSU6JRWK=
SW1783	NEH1VWdIem:5dHigbY5pcWKrdHnvckBie3OjeR?=		NF3OSG5KdmirYnn0bY9vKG:oIHj1cYFvKFOZMUe4N{Bk\WyuIHfyc5d1cCCrbjDhJINmdGxidnnhZoltcXS7IHHzd4F6NiCLQ{WwQVUzNjR7OEKg{txO	M37TbnNCVkeHUh?=
SBC-5	NF7aSWFIem:5dHigbY5pcWKrdHnvckBie3OjeR?=		MnH6TY5pcWKrdHnvckBw\iCqdX3hckBUSkNvNTDj[YxtKGe{b4f0bEBqdiCjIHPlcIwhfmmjYnnsbZR6KGG\|c3H5MkBKSzVyPUWxMlExQDVizszN	Mn35V2FPT0WU
A375	Mk\vS5Jwf3SqIHnubIljcXSrb36gZZN{[Xl?		NFHmdopKdmirYnn0bY9vKG:oIHj1cYFvKEF\|N{WgZ4VtdCCpcn;3eIghcW5iYTDj[YxtKH[rYXLpcIl1gSCjc4PhfU4hUUN3ME21N{43PzZ6IN88US=>	NF71OJNUSU6JRWK=
AM-38	M3HyPWdzd3e2aDDpcohq[mm2aX;uJIF{e2G7		Mn;NTY5pcWKrdHnvckBw\iCqdX3hckBCVS1\|ODDj[YxtKGe{b4f0bEBqdiCjIHPlcIwhfmmjYnnsbZR6KGG\|c3H5MkBKSzVyPUWyMlk1QTJizszN	MXrTRW5ITVJ?
YH-13	MV3Hdo94fGhiaX7obYJqfGmxbjDhd5NigQ>?		M1H3RmlvcGmkaYTpc44hd2ZiaIXtZY4hYUhvMUOgZ4VtdCCpcn;3eIghcW5iYTDj[YxtKH[rYXLpcIl1gSCjc4PhfU4hUUN3ME22NE4xPDZ6IN88US=>	MmD2V2FPT0WU
TYK-nu	NWLoTmxDT3Kxd4ToJIlvcGmkaYTpc44h[XO\|YYm=		NGLSW2pKdmirYnn0bY9vKG:oIHj1cYFvKFS\Sz3ueUBk\WyuIHfyc5d1cCCrbjDhJINmdGxidnnhZoltcXS7IHHzd4F6NiCLQ{WwQVc3NjZyNkOg{txO	M1j3TXNCVkeHUh?=
COLO-668	MmTuS5Jwf3SqIHnubIljcXSrb36gZZN{[Xl?		M2PvVWlvcGmkaYTpc44hd2ZiaIXtZY4hS0:OTz22Olgh[2WubDDndo94fGhiaX6gZUBk\WyuII\pZYJqdGm2eTDhd5NigS5iSVO1NF03OC53MEe3JO69VQ>?	NYLyfFJyW0GQR1XS
ES6	M2jVNmdzd3e2aDDpcohq[mm2aX;uJIF{e2G7		NULZRmg1UW6qaXLpeIlwdiCxZjDoeY1idiCHU{[gZ4VtdCCpcn;3eIghcW5iYTDj[YxtKH[rYXLpcIl1gSCjc4PhfU4hUUN3ME21O{44PTRzIN88US=>	MWrTRW5ITVJ?
D-247MG	MULHdo94fGhiaX7obYJqfGmxbjDhd5NigQ>?		NFjCbJlKdmirYnn0bY9vKG:oIHj1cYFvKERvMkS3UWch[2WubDDndo94fGhiaX6gZUBk\WyuII\pZYJqdGm2eTDhd5NigS5iSVO1NF02QS53OEi4JO69VQ>?	MkfRV2FPT0WU
H4	NVHRWnJIT3Kxd4ToJIlvcGmkaYTpc44h[XO\|YYm=		M1ThSGlvcGmkaYTpc44hd2ZiaIXtZY4hUDRiY3XscEBoem:5dHigbY4h[SClZXzsJJZq[WKrbHn0fUBie3OjeT6gTWM2OD14MD61OlE1KM7:TR?=	M1HYOHNCVkeHUh?=
HMV-II	NEnicHBIem:5dHigbY5pcWKrdHnvckBie3OjeR?=		NH\sWpFKdmirYnn0bY9vKG:oIHj1cYFvKEiPVj3JTUBk\WyuIHfyc5d1cCCrbjDhJINmdGxidnnhZoltcXS7IHHzd4F6NiCLQ{WwQVE2Py5zOUSg{txO	Mo\3V2FPT0WU
HT-1080	NGrhVZVIem:5dHigbY5pcWKrdHnvckBie3OjeR?=		M1fER2lvcGmkaYTpc44hd2ZiaIXtZY4hUFRvMUC4NEBk\WyuIHfyc5d1cCCrbjDhJINmdGxidnnhZoltcXS7IHHzd4F6NiCLQ{WwQVYzNjJyMzFOwG0>	M{\Q[3NCVkeHUh?=
IA-LM	NETCboFIem:5dHigbY5pcWKrdHnvckBie3OjeR?=		MVHJcohq[mm2aX;uJI9nKGi3bXHuJGlCNUyPIHPlcIwh\3Kxd4ToJIlvKGFiY3XscEB3cWGkaXzpeJkh[XO\|YYmuJGlEPTB;NUSuNFkyKM7:TR?=	NHXIVYNUSU6JRWK=
K-562	M1PVSGdzd3e2aDDpcohq[mm2aX;uJIF{e2G7		MnPMTY5pcWKrdHnvckBw\iCqdX3hckBMNTV4MjDj[YxtKGe{b4f0bEBqdiCjIHPlcIwhfmmjYnnsbZR6KGG\|c3H5MkBKSzVyPUWwMlU3OzNizszN	MXrTRW5ITVJ?
MEL-JUSO	MUDHdo94fGhiaX7obYJqfGmxbjDhd5NigQ>?		NYrTV49DUW6qaXLpeIlwdiCxZjDoeY1idiCPRVytTnVUVyClZXzsJIdzd3e2aDDpckBiKGOnbHygeoli[mmuaYT5JIF{e2G7LjDJR\|UxRTV4LkG3N\|ch\|ryP	MlXQV2FPT0WU
M059J	NUTJV4V{T3Kxd4ToJIlvcGmkaYTpc44h[XO\|YYm=		NUnqR\|RuUW6qaXLpeIlwdiCxZjDoeY1idiCPMEW5TkBk\WyuIHfyc5d1cCCrbjDhJINmdGxidnnhZoltcXS7IHHzd4F6NiCLQ{WwQVUyNjB{NUKg{txO	NGjZcnNUSU6JRWK=
NB6	MYLHdo94fGhiaX7obYJqfGmxbjDhd5NigQ>?		MWDJcohq[mm2aX;uJI9nKGi3bXHuJG5DPiClZXzsJIdzd3e2aDDpckBiKGOnbHygeoli[mmuaYT5JIF{e2G7LjDJR\|UxRTdzLkC3PVch\|ryP	MnT1V2FPT0WU
NCI-H1975	M1;QWHBzd2yrZnXyZZRqd25iYYPzZZk>	M2rXTFczKGh?	NVjqXJpSSW62aYDyc4xq\mW{YYTpeoUh[WO2aY\peJkh[WejaX7zeEBpfW2jbjDOR2kuUDF7N{WgZ4VtdHNiZYjwdoV{e2mwZzDFS2ZTKFR5OUDNJI12fGGwdDDh[pRmeiB5MjDodpMh[nliU2LCJIF{e2G7LDDJR\|UxRTBwMUS4JO69VQ>?	M1zTflI1OTh\|N{Sy
A431	NHXtcXNRem:uaX\ldoF1cW:wIHHzd4F6	M{Wy[lczKGh?	NGDCbGtCdnSrcILvcIln\XKjdHn2[UBi[3Srdnn0fUBi\2GrboP0JIh2dWGwIFG0N\|Eh[2WubIOgZYZ1\XJiN{KgbJJ{KGK7IGPSRkBie3OjeTygTWM2OD1yLkCyN{BvVQ>?	M12yN\|I1OTh\|N{Sy
MDA-MB-231	NXzE[ZFtWHKxbHnm[ZJifGmxbjDhd5NigQ>?	NXPl[FZoPzJiaB?=	NYr4c5FMSW62aYDyc4xq\mW{YYTpeoUh[WO2aY\peJkh[WejaX7zeEBpfW2jbjDNSGEuVUJvMkOxJINmdGy\|IHHmeIVzKDd{IHjyd{BjgSCPVGSgZZN{[XluIFXDOVA:OSCwTR?=	MmjmNlAyQDB7ME[=
NCI-H1975	M{TSdWN6fG:2b4jpZ4l1gSCjc4PhfS=>	MWq0PEBp	NYjvd21sS3m2b4TvfIlkcXS7IHHnZYlve3RiaIXtZY4hVkOLLVixPVc2KGOnbHzzJIFnfGW{IES4JIhzeyCkeTDNWHQh[XO\|YYmsJGlEPTB;MD6zPVIh\|ryP	M4TEXFIxOTVyNEC0
NIH/3T3	MkCyR5l1d3SxeHnjbZR6KGG\|c3H5	MXW0PEBp	MkLmR5l1d3SxeHnjbZR6KGGpYXnud5QhdW:3c3WgUmlJNzOWMzDj[YxteyCjZoTldkA1QCCqcoOgZpkhVVSWIHHzd4F6NCCLQ{WwQVMvPjB3IN88US=>	MVSyNFE2ODRyNB?=
A549	NHfBeIxEgXSxdH;4bYNqfHliYYPzZZk>	NEnsWIw4OiCq	M{H6OWN6fG:2b4jpZ4l1gSCjZ3HpcpN1KGi3bXHuJGE2PDliY3XscJMh[W[2ZYKgO\|IhcHK\|IHL5JG1VXCCjc4PhfUwhUUN3ME2wMlA2KM7:TR?=	NVL1TJZ4OjBzOEC5NFU>
PC3	NH7DbVBEgXSxdH;4bYNqfHliYYPzZZk>	MWm3NkBp	MXHDfZRwfG:6aXPpeJkh[WejaX7zeEBpfW2jbjDQR\|Mh[2WubIOgZYZ1\XJiN{KgbJJ{KGK7IF3UWEBie3OjeTygTWM2OD12LkGg{txO	MVWyNFE5ODlyNR?=
HeLa	MoPrR5l1d3SxeHnjbZR6KGG\|c3H5	NUPJfoVbPzJiaB?=	NXXOVoZ[S3m2b4TvfIlkcXS7IHHnZYlve3RiaIXtZY4hUGWOYTDj[YxteyCjZoTldkA4OiCqcoOgZpkhVVSWIHHzd4F6NCCLQ{WwQVYvQDFizszN	M13HUVIxOThyOUC1
MCF7	M1nDbGN6fG:2b4jpZ4l1gSCjc4PhfS=>	NVXuS\|c2PzJiaB?=	NXPJUnp{S3m2b4TvfIlkcXS7IHHnZYlve3RiaIXtZY4hVUOINzDj[YxteyCjZoTldkA4OiCqcoOgZpkhVVSWIHHzd4F6NCCLQ{WwQVUvQDNizszN	MXSyNFE5ODlyNR?=
NSCLC	NH;Y[ndEgXSxdH;4bYNqfHliYYPzZZk>	M{nxVVczKGh?	MULDfZRwfG:6aXPpeJkh[WejaX7zeEBpfW2jbjDOV2NNSyClZXzsd{whUUN3ME2wMlUhdk1?	NFXWXWIzODF6MEmwOS=>

... Click to View More Cell Line Experimental Data

Assay

Methods	Test Index	PMID
Growth inhibition assay	Cell apoptosis; PubMed: 25537503 Oncotarget Dose-dependent effects of afatinib on apoptosis. Data are mean ± SD. n = 3 for each concentration. , p < 0.05, *, p < 0.01, vs. no afatinib. Cell viability; PubMed: 26515464 Oncotarget A. MTS assay for Ba/F3 cells harboring EGFR exon 19 deletion and L858R. The proportional cell viability is shown. B. MTS assay for Ba/F3 cells harboring EGFR exon 19 deletion+T790M and L858R+T790M. The proportional cell viability is shown. C. MTS assay for Ba/F3 cells harboring wild type EGFR. The proportional cell viability is shown. Erlotinib, afatinib, osimertinib, and rociletinib were used as EGFR-TKIs. Error bars indicate standard deviation.	25537503 26515464
Western blot	CIP2A / p-AKT / AKT / PARP; PubMed: 25537503 Oncotarget Time-dependent effects of afatinib on CIP2A, p-AKT and apoptosis-related proteins in the sensitive H358 and resistant H460 cells. Cells were exposed to 10 μM afatinib for up to 48 hours. Immunoblots were scanned and quantitated to determine the ratio of the level of CIP2A to actin. Data are mean ± SD. n = 3 for the different time intervals. p-EGFR / p-IGF1R / p-ERK / p-AKT; PubMed: 26052929 Mol Carcinog Effects of afatinib on EGFR and IGF1R Signaling Pathways. H1975 cells were treated with afatinib with various doses and cell lysates were subjected to Western blot analysis. Afatinib effectively inhibits p-EGFR and p-ERK while p-AKT was partially inhibited. IGF1R phosphorylation was increased upon afatinib treatment in H1975 cells.	25537503 26052929
Immunofluorescence	p65 ; PubMed: 26317651 Oncotarget Effects of different treatments on the nuclear translocation of the NF-κB subunit p65. A2780T cells were treated with 2.5 μM afatinib for 48 hours, or 10 μM PDTC for 2 hours. Subsequently, the NF-κB subunit p65 (red) was localized by immunofluorescence and a confocal microscopy. Nuclei were stained with DAPI (blue). Scale bar = 10 μm. vacuoles; PubMed: 29713246 Cancer Cell Int Autophagy activation upon treatment with Afatinib and Sorafenib was validated using immunofluorescence in order to visualize a creation of autophagosomes; ×60 magnification. DMSO treatment was used as control. Green—autophagy vacuoles; Blue—Hoechst, nuclear staining.	26317651 29713246

In vivo

In the MDA-MB-453 xenograft model, BIBW2992 (20 mg/kg, p.o.) results in dramatic tumor regression with a cumulative treated/control tumor volume ratio (T/C ratio) of 2%, and downregulation of EGFR and AKT phosphorylation.^[1] In A7, A431, FaDu, UT-SCC-14 and UT-SCC-15 xenograft models, application of BIBW 2992 (30 mg/kg, p.o.) leads to a significant prolongation of tumour growth time.^[3] In HER2-amplified xenograft medel, Afatinib (30 mg/kg, p.o.) results a markedly inhibition in tumor growth and a significant improvement in the duration of overall survival.^[4] In HER2-positive gastric cancer NCI-N87 xenograft, afatinib (25 mg/kg, p.o.) leads to dramatic tumor volume regression within 4 d and near-complete tumor resolution after 21 d of treatment.^[5]

Protocol

Kinase Assay:^[1]	- Collapse In vitro kinase activity assay: EGFR kinase: Each 100 µL enzyme reaction contained 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. The enzymatic reaction is started by addition of 50 µL of a 100 µM ATP solution made in 10 mM MgCl2. Assays are carried out at room temperature for 30 min and terminated by the addition of 50 µL of stop solution (250 mM EDTA in 20 mM HEPES pH 7.4). 100 µL are transferred to a streptavidin coated microtiterplate, after an incubation time of 60 min at room temperature the plate is washed with 200 µL of wash solution (50 mM Tris, 0.05% Tween20). A 100 µL aliquot of a HRPO- labeled anti-PY antibody (PY20H Anti-Ptyr:HRP ) 250 ng/mL are added to the wells. After 60 min of incubation, the plate is washed three times with a 200 µL wash solution. The samples are then developed with a 100µL TMB Peroxidase Solution (A:B= 1:1). The reaction is stopped after 10 min. The plate is transferred to an ELISA reader and extinction is measured at OD450nm. HER2-IC enzyme: Enzyme activity is assayed in the presence or absence of serial inhibitor dilutions performed in 50 % Me2SO. Each 100 µL reaction contains similar components as described for EGFR kinase assay with addition of 1000 µM Na3VO4. The enzymatic reaction is started by addition of 50µL of 500 µM ATP solution made in 10 mM Mg-acetate. The dilution of the enzyme is set so that incorporation of phosphate into bio-pEY is linear with respect to time and amount of enzyme. The enzyme preparation is diluted in 20 mM HEPES pH 7.4, 130 mM NaCl, 0.05% Triton X-100, 1 mM DTT and 10% glycerol. Assays are carried out at room temperature for 30 min and terminated by the addition of 50 µL of stop solution. Src kinase assays: Each 100 µL reaction contained 10 µL of inhibitor in 50 % Me2SO, 20µL of enzyme preparation, 20 µL of substrate solution supplemented with 1000 µM Na3VO4. The enzymatic reaction is started by addition of 50 µL of a 1000 µM ATP solution made in 10 mM Mg-acetate. BIRK kinase assay: 250 mM Tris pH 7.4, 10mM DTT, 2.5 mg/mL poly(EY), 5 mg/mL bio-pEY is used as substrate solution and enzymatic reaction is started by addition of 50 µL of a 2 mM ATP solution made in 8 mM MnCl2, 20 mM Mg-acetate. VEGF2 and HGFR kinase assays: Assays are carried out at room temperature for 20 minutes and terminated by the addition of 10 µL of 5 % H3PO4. The precipitate is then trapped onto GF/B filters using a 96 well filter mate universal harvester. After extensive washing the filter plate is dried for 1 h at 50°C, sealed and incorporated radioactivity is determined by scintillation counting using a TopCount™ or a Microbeta b counter™.
Cell Research:^[2]	- Collapse Cell lines: SP and NSP cell lines Concentrations: 1 μM Incubation Time: 48 h Method: Cytotoxicity is determined using MTT assay. The IC 50 value is deﬁned as the drug concentration resulting in 50% cell death. Both the ﬁtted sigmoidal dose response curve and IC50 are calculated by Bliss method. (Only for Reference)
Animal Research:^[4]	- Collapse Animal Models: SCID mice harbouring ARK2 xenografts Dosages: 25 mg/kg Administration: p.o. (Only for Reference)

References

- Collapse

Solubility (25°C)

In vitro	DMSO	100 mg/mL (139.43 mM)
	Water	Insoluble
	Ethanol	Insoluble
In vivo	Add solvents to the product individually and in order(Data is from Selleck tests instead of citations): 5% DMSO+30% PEG 300+ddH2O For best results, use promptly after mixing.	28mg/mL

* 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 Information Download Afatinib (BIBW2992) Dimaleate SDF

Molecular Weight	717.18
Formula	C₂₄H₂₅ClFN₅O₃.2C₄H₄O₄
CAS No.	850140-73-7
Storage	3 years-20°C powder 2 years-80°C in solvent
Synonyms	N/A
Smiles	CN(C)CC=CC(=O)NC1=C(C=C2C(=C1)C(=NC=N2)NC3=CC(=C(C=C3)F)Cl)OC4CCOC4.C(=CC(=O)O)C(=O)O.C(=CC(=O)O)C(=O)O

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% DMSO+ % + % Tween 80+ % ddH₂O

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Working concentration： mg/ml；

Method for preparing DMSO master liquid: ： mg drug pre-dissolved in μL DMSO (Master liquid 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 master liquid, next addμL PEG300， mix and clarify, next addμL Tween 80，mix and clarify, next add μL ddH₂O，mix and clarify.

Note：1.Please make sure the liquid is clear before adding the next solvent.
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The Serial Dilution Calculator Equation

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Computed Result

C1=C0/X	C1:	LOG(C1):
C2=C1/X	C2:	LOG(C2):
C3=C2/X	C3:	LOG(C3):
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Molecular Weight Calculator

Enter the chemical formula of a compound to calculate its molar mass and elemental composition:

Tip: Chemical formula is case sensitive. C10H16N2O2 c10h16n2o2

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Molarity Calculator

Clinical Trial Information (data from https://clinicaltrials.gov, updated on 2021-09-06)

NCT Number	Recruitment	interventions	Conditions	Sponsor/Collaborators	Start Date	Phases
NCT04497584	Not yet recruiting	Drug: Afatinib + Prednisone	Advanced Squamous Non Small Cell Lung Cancer	University of Texas Southwestern Medical Center	September 1 2021	Phase 2
NCT04795245	Not yet recruiting	Drug: afatinib	Non-squamous Non-Small Cell Lung Cancer	Boehringer Ingelheim	August 31 2021	--
NCT04413201	Recruiting	Drug: Afatinib\|Drug: Osimertinib	Non-squamous NSCLC	Michael Hopp\|Boehringer Ingelheim\|Johannes Gutenberg University Mainz	September 11 2020	Phase 4
NCT04206787	Recruiting	Drug: Afatinib	Non-squamous Non-Small Cell Lung Cancer	Boehringer Ingelheim	May 20 2020	--
NCT04158947	Recruiting	Drug: Afatinib\|Drug: T-DM1	HER2-positive Breast Cancer\|Brain Metastases	xuexin he\|Second Affiliated Hospital School of Medicine Zhejiang University	May 10 2020	Phase 2

Tech Support

Answers to questions you may have can be found in the inhibitor handling instructions. Topics include how to prepare stock solutions, how to store inhibitors, and issues that need special attention for cell-based assays and animal experiments.

Handling Instructions

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EGFR Signaling Pathway Map

Related EGFR Products

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Poziotinib (HM781-36B) ^New

Poziotinib (HM781-36B, NOV120101) is an irreversible pan-HER inhibitor with IC50 of 3.2 nM, 5.3 nM and 23.5 nM for HER1, HER2, and HER4, respectively. Poziotinib also induces apoptosis and G1 cell cycle arrest. Phase 2.
AZ5104 ^New

AZ5104, the demethylated metabolite of AZD-9291, is a potent EGFR inhibitor with IC50 of <1 nM, 6 nM, 1 nM, and 25 nM for EGFR (L858R/T790M), EGFR (L858R), EGFR (L861Q), and EGFR (wildtype), respectively. Phase 1.
Gefitinib (ZD1839)

Gefitinib (ZD-1839, Iressa) is an EGFR inhibitor for Tyr1173, Tyr992, Tyr1173 and Tyr992 in the NR6wtEGFR and NR6W cells with IC50 of 37 nM, 37nM, 26 nM and 57 nM, respectively. Gefitinib promotes autophagy and apoptosis of lung cancer cells via blockade of the PI3K/AKT/mTOR pathway.

Features:A potent EGFR tyrosine kinase inhibitor.
Erlotinib HCl (OSI-744)

Erlotinib HCl (OSI-744, CP358774, NSC 718781) is an EGFR inhibitor with IC50 of 2 nM in cell-free assays, >1000-fold more sensitive for EGFR than human c-Src or v-Abl.
Afatinib (BIBW2992)

Afatinib (BIBW2992) inhibits EGFR/ErbB irreversibly in vitro with IC50 of 0.5, 0.4, 10, 14, 1 nM for EGFR^wt, EGFR^L858R, EGFR^L858R/T790M ErbB2 (HER2) and ErbB4 (HER4), respectively. Afatinib induces autophagy.
Osimertinib (AZD9291)

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Lapatinib (GW-572016, GSK572016, GW2016), used in the form of Lapatinib Ditosylate, is a potent EGFR and ErbB2 inhibitor with IC50 of 10.8 and 9.2 nM in cell-free assays, respectively. Lapatinib induces ferroptosis and autophagic cell death.
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AG-490 (Tyrphostin B42, Zinc02557947) is an inhibitor of EGFR with IC50 of 0.1 μM in cell-free assays, 135-fold more selective for EGFR versus ErbB2, also inhibits JAK2 with no activity to Lck, Lyn, Btk, Syk and Src.
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Afatinib (BIBW2992) Dimaleate