Afatinib (BIBW2992) Dimaleate

Catalog No.S7810

For research use only.

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.

Afatinib (BIBW2992) Dimaleate Chemical Structure

CAS No. 850140-73-7

Selleck's Afatinib (BIBW2992) Dimaleate has been cited by 134 publications

Purity & Quality Control

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Biological Activity

Description 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.
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 Concentration Incubation Time Formulation Activity Description PMID
HCC827 cell NV\5W|FoWHKxbHnm[ZJifGmxbjDhd5NigQ>? NVP6O5dISW62aYDyc4xq\mW{YYTpeoUh[WO2aY\peJkh[WejaX7zeEBpfW2jbjDIR2M5OjdiY3XscJMh[W[2ZYKgPVYhcHK|IHL5JG1VWyCjc4PhfUwhUUN3ME24JI5O MnHLNlA2PTB{MUK=
NCI-H1563 NEjBWJdIem:5dHigbY5pcWKrdHnvckBie3OjeR?= MYrJcohq[mm2aX;uJI9nKGi3bXHuJG5EUS2KMUW2N{Bk\WyuIHfyc5d1cCCrbjDhJINmdGxidnnhZoltcXS7IHHzd4F6NiCLQ{WwQVkvPzZyM{Gg{txO NUG4SJRVW0GQR1XS
NCI-H1573 NFvSd|hIem:5dHigbY5pcWKrdHnvckBie3OjeR?= NXvRTII{UW6qaXLpeIlwdiCxZjDoeY1idiCQQ1mtTFE2PzNiY3XscEBoem:5dHigbY4h[SClZXzsJJZq[WKrbHn0fUBie3OjeT6gTWM2OD1yLkezOlMh|ryP NXTOXGg2W0GQR1XS
NCI-H1581 NIS1V4VIem:5dHigbY5pcWKrdHnvckBie3OjeR?= MlnVTY5pcWKrdHnvckBw\iCqdX3hckBPS0lvSEG1PFEh[2WubDDndo94fGhiaX6gZUBk\WyuII\pZYJqdGm2eTDhd5NigS5iSVO1NF0zQS5|MUS1JO69VQ>? NIH4RYRUSU6JRWK=
NCI-H1648 NH\K[XVIem:5dHigbY5pcWKrdHnvckBie3OjeR?= M3rDd2lvcGmkaYTpc44hd2ZiaIXtZY4hVkOLLVixOlQ5KGOnbHyg[5Jwf3SqIHnuJIEh[2WubDD2bYFjcWyrdImgZZN{[XlwIFnDOVA:OC52NEG4NkDPxE1? NWHsUpdWW0GQR1XS
NCI-H1793 M1;XO2dzd3e2aDDpcohq[mm2aX;uJIF{e2G7 MlP1TY5pcWKrdHnvckBw\iCqdX3hckBPS0lvSEG3PVMh[2WubDDndo94fGhiaX6gZUBk\WyuII\pZYJqdGm2eTDhd5NigS5iSVO1NF0yPC5{MEi3JO69VQ>? MX7TRW5ITVJ?
NCI-H1838 NGPGO4tIem:5dHigbY5pcWKrdHnvckBie3OjeR?= NW\vO21IUW6qaXLpeIlwdiCxZjDoeY1idiCQQ1mtTFE5OzhiY3XscEBoem:5dHigbY4h[SClZXzsJJZq[WKrbHn0fUBie3OjeT6gTWM2OD1yLk[wNlk6KM7:TR?= Mon0V2FPT0WU
NCI-H1770 NVXDfWx7T3Kxd4ToJIlvcGmkaYTpc44h[XO|YYm= Mk\STY5pcWKrdHnvckBw\iCqdX3hckBPS0lvSEG3O|Ah[2WubDDndo94fGhiaX6gZUBk\WyuII\pZYJqdGm2eTDhd5NigS5iSVO1NF0xNjFyMkig{txO NWPFZ3BiW0GQR1XS
NCI-H1651 M13Pe2dzd3e2aDDpcohq[mm2aX;uJIF{e2G7 NWO5WWdyUW6qaXLpeIlwdiCxZjDoeY1idiCQQ1mtTFE3PTFiY3XscEBoem:5dHigbY4h[SClZXzsJJZq[WKrbHn0fUBie3OjeT6gTWM2OD1{NT61PVch|ryP NGDvXm9USU6JRWK=
NCI-H1975 M2C0Wmdzd3e2aDDpcohq[mm2aX;uJIF{e2G7 NFnMXWxKdmirYnn0bY9vKG:oIHj1cYFvKE6FST3INVk4PSClZXzsJIdzd3e2aDDpckBiKGOnbHygeoli[mmuaYT5JIF{e2G7LjDJR|UxRTBwNkG3Nlgh|ryP NFLZT3hUSU6JRWK=
NCI-H1666 MX3Hdo94fGhiaX7obYJqfGmxbjDhd5NigQ>? MlPDTY5pcWKrdHnvckBw\iCqdX3hckBPS0lvSEG2OlYh[2WubDDndo94fGhiaX6gZUBk\WyuII\pZYJqdGm2eTDhd5NigS5iSVO1NF0yNjhyOUS4JO69VQ>? MkPXV2FPT0WU
NCI-H2009 NWfYOHlqT3Kxd4ToJIlvcGmkaYTpc44h[XO|YYm= NYHCeGF[UW6qaXLpeIlwdiCxZjDoeY1idiCQQ1mtTFIxODliY3XscEBoem:5dHigbY4h[SClZXzsJJZq[WKrbHn0fUBie3OjeT6gTWM2OD1|Lk[xNlE3KM7:TR?= NInKfGhUSU6JRWK=
NCI-H2126 M4LBUmdzd3e2aDDpcohq[mm2aX;uJIF{e2G7 Mn;VTY5pcWKrdHnvckBw\iCqdX3hckBPS0lvSEKxNlYh[2WubDDndo94fGhiaX6gZUBk\WyuII\pZYJqdGm2eTDhd5NigS5iSVO1NF0xNjVzOUK2JO69VQ>? MWrTRW5ITVJ?
NCI-H1693 MmHsS5Jwf3SqIHnubIljcXSrb36gZZN{[Xl? NEHMRXpKdmirYnn0bY9vKG:oIHj1cYFvKE6FST3INVY6OyClZXzsJIdzd3e2aDDpckBiKGOnbHygeoli[mmuaYT5JIF{e2G7LjDJR|UxRTNwOESxPFEh|ryP NX22UpF3W0GQR1XS
NCI-H2087 NXnIR|FCT3Kxd4ToJIlvcGmkaYTpc44h[XO|YYm= MlTKTY5pcWKrdHnvckBw\iCqdX3hckBPS0lvSEKwPFch[2WubDDndo94fGhiaX6gZUBk\WyuII\pZYJqdGm2eTDhd5NigS5iSVO1NF01QS57NkiyJO69VQ>? NF23T4dUSU6JRWK=
NCI-H2029 NX7wbXpuT3Kxd4ToJIlvcGmkaYTpc44h[XO|YYm= NIXGPZZKdmirYnn0bY9vKG:oIHj1cYFvKE6FST3INlAzQSClZXzsJIdzd3e2aDDpckBiKGOnbHygeoli[mmuaYT5JIF{e2G7LjDJR|UxRTR|LkW1PFEh|ryP MkD3V2FPT0WU
NCI-H2170 NHSyZZBIem:5dHigbY5pcWKrdHnvckBie3OjeR?= MmjGTY5pcWKrdHnvckBw\iCqdX3hckBPS0lvSEKxO|Ah[2WubDDndo94fGhiaX6gZUBk\WyuII\pZYJqdGm2eTDhd5NigS5iSVO1NF0xNjB|N{K4JO69VQ>? M4PL[HNCVkeHUh?=
NCI-H226 NEHzTGlIem:5dHigbY5pcWKrdHnvckBie3OjeR?= NHzXXY1KdmirYnn0bY9vKG:oIHj1cYFvKE6FST3INlI3KGOnbHyg[5Jwf3SqIHnuJIEh[2WubDD2bYFjcWyrdImgZZN{[XlwIFnDOVA:PDlwN{e4OUDPxE1? M2rO[XNCVkeHUh?=
NCI-H2030 NEfZeVRIem:5dHigbY5pcWKrdHnvckBie3OjeR?= MmHHTY5pcWKrdHnvckBw\iCqdX3hckBPS0lvSEKwN|Ah[2WubDDndo94fGhiaX6gZUBk\WyuII\pZYJqdGm2eTDhd5NigS5iSVO1NF0yPC54NES4JO69VQ>? M3jWdHNCVkeHUh?=
NCI-H2052 NIHmRoRIem:5dHigbY5pcWKrdHnvckBie3OjeR?= MYrJcohq[mm2aX;uJI9nKGi3bXHuJG5EUS2KMkC1NkBk\WyuIHfyc5d1cCCrbjDhJINmdGxidnnhZoltcXS7IHHzd4F6NiCLQ{WwQVIyNjh7NkOg{txO MkfmV2FPT0WU
NCI-H292 MVjHdo94fGhiaX7obYJqfGmxbjDhd5NigQ>? MYHJcohq[mm2aX;uJI9nKGi3bXHuJG5EUS2KMkmyJINmdGxiZ4Lve5RpKGmwIHGgZ4VtdCC4aXHibYxqfHliYYPzZZkvKEmFNUC9NE4xPjB7IN88US=> MkHVV2FPT0WU
NCI-H441 NVPiW2ZMT3Kxd4ToJIlvcGmkaYTpc44h[XO|YYm= M2TWXGlvcGmkaYTpc44hd2ZiaIXtZY4hVkOLLVi0OFEh[2WubDDndo94fGhiaX6gZUBk\WyuII\pZYJqdGm2eTDhd5NigS5iSVO1NF0{NjV7OESxJO69VQ>? MlvDV2FPT0WU
NCI-H2291 NH:yV2dIem:5dHigbY5pcWKrdHnvckBie3OjeR?= NIO5WWlKdmirYnn0bY9vKG:oIHj1cYFvKE6FST3INlI6OSClZXzsJIdzd3e2aDDpckBiKGOnbHygeoli[mmuaYT5JIF{e2G7LjDJR|UxRTVwOUi5OFMh|ryP M2DrOHNCVkeHUh?=
NCI-H358 NWnRNYFQT3Kxd4ToJIlvcGmkaYTpc44h[XO|YYm= NFTNb3dKdmirYnn0bY9vKG:oIHj1cYFvKE6FST3IN|U5KGOnbHyg[5Jwf3SqIHnuJIEh[2WubDD2bYFjcWyrdImgZZN{[XlwIFnDOVA:OC53M{KxJO69VQ>? M1zjb3NCVkeHUh?=
NCI-H460 Ml\VS5Jwf3SqIHnubIljcXSrb36gZZN{[Xl? MkHITY5pcWKrdHnvckBw\iCqdX3hckBPS0lvSES2NEBk\WyuIHfyc5d1cCCrbjDhJINmdGxidnnhZoltcXS7IHHzd4F6NiCLQ{WwQVQ3NjV{MzFOwG0> NX7RbmZwW0GQR1XS
NCI-H661 M3jiPGdzd3e2aDDpcohq[mm2aX;uJIF{e2G7 MlLSTY5pcWKrdHnvckBw\iCqdX3hckBPS0lvSE[2NUBk\WyuIHfyc5d1cCCrbjDhJINmdGxidnnhZoltcXS7IHHzd4F6NiCLQ{WwQVE3Njl4OUSg{txO MYjTRW5ITVJ?
NCI-H23 Mo\hS5Jwf3SqIHnubIljcXSrb36gZZN{[Xl? M13ob2lvcGmkaYTpc44hd2ZiaIXtZY4hVkOLLViyN{Bk\WyuIHfyc5d1cCCrbjDhJINmdGxidnnhZoltcXS7IHHzd4F6NiCLQ{WwQVM4NjRzOU[g{txO MXTTRW5ITVJ?
NCI-H630 NGXkcIVIem:5dHigbY5pcWKrdHnvckBie3OjeR?= MmPlTY5pcWKrdHnvckBw\iCqdX3hckBPS0lvSE[zNEBk\WyuIHfyc5d1cCCrbjDhJINmdGxidnnhZoltcXS7IHHzd4F6NiCLQ{WwQVEvPDV3OEKg{txO NWTOXmJYW0GQR1XS
NCI-H2342 NYHVUmRRT3Kxd4ToJIlvcGmkaYTpc44h[XO|YYm= MUjJcohq[mm2aX;uJI9nKGi3bXHuJG5EUS2KMkO0NkBk\WyuIHfyc5d1cCCrbjDhJINmdGxidnnhZoltcXS7IHHzd4F6NiCLQ{WwQVE4NjV3NzFOwG0> NYDqe|R2W0GQR1XS
NCI-H69 NET6TGNIem:5dHigbY5pcWKrdHnvckBie3OjeR?= MUXJcohq[mm2aX;uJI9nKGi3bXHuJG5EUS2KNkmgZ4VtdCCpcn;3eIghcW5iYTDj[YxtKH[rYXLpcIl1gSCjc4PhfU4hUUN3ME2wMlIyPDVzIN88US=> M1PtOHNCVkeHUh?=
NCI-H650 MYDHdo94fGhiaX7obYJqfGmxbjDhd5NigQ>? NXrLdml2UW6qaXLpeIlwdiCxZjDoeY1idiCQQ1mtTFY2OCClZXzsJIdzd3e2aDDpckBiKGOnbHygeoli[mmuaYT5JIF{e2G7LjDJR|UxRTFzLkK0NVgh|ryP M3fi[3NCVkeHUh?=
NCI-H747 Mlm3S5Jwf3SqIHnubIljcXSrb36gZZN{[Xl? MX\Jcohq[mm2aX;uJI9nKGi3bXHuJG5EUS2KN{S3JINmdGxiZ4Lve5RpKGmwIHGgZ4VtdCC4aXHibYxqfHliYYPzZZkvKEmFNUC9OU41OjR5MjFOwG0> NH7OU5VUSU6JRWK=
NCI-H2405 NVq1SolJT3Kxd4ToJIlvcGmkaYTpc44h[XO|YYm= NFLuWItKdmirYnn0bY9vKG:oIHj1cYFvKE6FST3INlQxPSClZXzsJIdzd3e2aDDpckBiKGOnbHygeoli[mmuaYT5JIF{e2G7LjDJR|UxRTF5LkG5O|Qh|ryP MW\TRW5ITVJ?
NCI-H520 NWXPb45JT3Kxd4ToJIlvcGmkaYTpc44h[XO|YYm= MnLhTY5pcWKrdHnvckBw\iCqdX3hckBPS0lvSEWyNEBk\WyuIHfyc5d1cCCrbjDhJINmdGxidnnhZoltcXS7IHHzd4F6NiCLQ{WwQVI1Njh6NkGg{txO NUDDPIxKW0GQR1XS
NCI-H810 NYDNWVVLT3Kxd4ToJIlvcGmkaYTpc44h[XO|YYm= NIrmXYRKdmirYnn0bY9vKG:oIHj1cYFvKE6FST3IPFExKGOnbHyg[5Jwf3SqIHnuJIEh[2WubDD2bYFjcWyrdImgZZN{[XlwIFnDOVA:OzRwN{K3PEDPxE1? M4HDUHNCVkeHUh?=
NCI-H720 M1eyTmdzd3e2aDDpcohq[mm2aX;uJIF{e2G7 M1izVGlvcGmkaYTpc44hd2ZiaIXtZY4hVkOLLVi3NlAh[2WubDDndo94fGhiaX6gZUBk\WyuII\pZYJqdGm2eTDhd5NigS5iSVO1NF0xNjB3M{K1JO69VQ>? MV7TRW5ITVJ?
NCI-SNU-1 MUfHdo94fGhiaX7obYJqfGmxbjDhd5NigQ>? Mnf1TY5pcWKrdHnvckBw\iCqdX3hckBPS0lvU17VMVEh[2WubDDndo94fGhiaX6gZUBk\WyuII\pZYJqdGm2eTDhd5NigS5iSVO1NF0{NjZ7N{O1JO69VQ>? NY\TXpNwW0GQR1XS
NH-12 M1LrO2dzd3e2aDDpcohq[mm2aX;uJIF{e2G7 M1nkWWlvcGmkaYTpc44hd2ZiaIXtZY4hVkhvMUKgZ4VtdCCpcn;3eIghcW5iYTDj[YxtKH[rYXLpcIl1gSCjc4PhfU4hUUN3ME23MlczQDB6IN88US=> M{XJN3NCVkeHUh?=
NCI-SNU-5 MX;Hdo94fGhiaX7obYJqfGmxbjDhd5NigQ>? NW\XZ|NPUW6qaXLpeIlwdiCxZjDoeY1idiCQQ1mtV25WNTViY3XscEBoem:5dHigbY4h[SClZXzsJJZq[WKrbHn0fUBie3OjeT6gTWM2OD1zLkKzPUDPxE1? M4XL[HNCVkeHUh?=
NKM-1 NVXyOGNqT3Kxd4ToJIlvcGmkaYTpc44h[XO|YYm= M4rUXmlvcGmkaYTpc44hd2ZiaIXtZY4hVkuPLUGgZ4VtdCCpcn;3eIghcW5iYTDj[YxtKH[rYXLpcIl1gSCjc4PhfU4hUUN3ME2zMlE5PzR2IN88US=> M2jUUXNCVkeHUh?=
NCI-H526 MnLtS5Jwf3SqIHnubIljcXSrb36gZZN{[Xl? NXrNW4lQUW6qaXLpeIlwdiCxZjDoeY1idiCQQ1mtTFUzPiClZXzsJIdzd3e2aDDpckBiKGOnbHygeoli[mmuaYT5JIF{e2G7LjDJR|UxRTBwMEe4NlMh|ryP MVrTRW5ITVJ?
NTERA-S-cl-D1 MXjHdo94fGhiaX7obYJqfGmxbjDhd5NigQ>? NF3RPXdKdmirYnn0bY9vKG:oIHj1cYFvKE6WRWLBMXMu[2xvREGgZ4VtdCCpcn;3eIghcW5iYTDj[YxtKH[rYXLpcIl1gSCjc4PhfU4hUUN3ME21MlU1PTN5IN88US=> MYfTRW5ITVJ?
NMC-G1 MnvsS5Jwf3SqIHnubIljcXSrb36gZZN{[Xl? MnOxTY5pcWKrdHnvckBw\iCqdX3hckBPVUNvR{GgZ4VtdCCpcn;3eIghcW5iYTDj[YxtKH[rYXLpcIl1gSCjc4PhfU4hUUN3ME2xNk46OjN2IN88US=> NGn3RZFUSU6JRWK=
NCI-H82 M3j6V2dzd3e2aDDpcohq[mm2aX;uJIF{e2G7 MV\Jcohq[mm2aX;uJI9nKGi3bXHuJG5EUS2KOEKgZ4VtdCCpcn;3eIghcW5iYTDj[YxtKH[rYXLpcIl1gSCjc4PhfU4hUUN3ME2xMlU1ODB4IN88US=> MXzTRW5ITVJ?
NCI-H1623 NYO0UXJtT3Kxd4ToJIlvcGmkaYTpc44h[XO|YYm= M37jcWlvcGmkaYTpc44hd2ZiaIXtZY4hVkOLLVixOlI{KGOnbHyg[5Jwf3SqIHnuJIEh[2WubDD2bYFjcWyrdImgZZN{[XlwIFnDOVA:OC5yOECyOUDPxE1? NXzsNYt2W0GQR1XS
NOMO-1 Mke1S5Jwf3SqIHnubIljcXSrb36gZZN{[Xl? M1SzdmlvcGmkaYTpc44hd2ZiaIXtZY4hVk:PTz2xJINmdGxiZ4Lve5RpKGmwIHGgZ4VtdCC4aXHibYxqfHliYYPzZZkvKEmFNUC9N|QvPTdyMzFOwG0> NH;SWo5USU6JRWK=
NCI-N87 MWnHdo94fGhiaX7obYJqfGmxbjDhd5NigQ>? NIrEXZZKdmirYnn0bY9vKG:oIHj1cYFvKE6FST3OPFch[2WubDDndo94fGhiaX6gZUBk\WyuII\pZYJqdGm2eTDhd5NigS5iSVO1NF0xNjB{N{ezJO69VQ>? MYrTRW5ITVJ?
NCI-H1650 Mn;1S5Jwf3SqIHnubIljcXSrb36gZZN{[Xl? MmryTY5pcWKrdHnvckBw\iCqdX3hckBPS0lvSEG2OVAh[2WubDDndo94fGhiaX6gZUBk\WyuII\pZYJqdGm2eTDhd5NigS5iSVO1NF06NjZyOUS2JO69VQ>? MUHTRW5ITVJ?
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NCI-H209 M2O5dGdzd3e2aDDpcohq[mm2aX;uJIF{e2G7 NVPQd3lzUW6qaXLpeIlwdiCxZjDoeY1idiCQQ1mtTFIxQSClZXzsJIdzd3e2aDDpckBiKGOnbHygeoli[mmuaYT5JIF{e2G7LjDJR|UxRTBwMkC0OkDPxE1? Ml6xV2FPT0WU
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NY MnewS5Jwf3SqIHnubIljcXSrb36gZZN{[Xl? M{flfmlvcGmkaYTpc44hd2ZiaIXtZY4hVlliY3XscEBoem:5dHigbY4h[SClZXzsJJZq[WKrbHn0fUBie3OjeT6gTWM2OD1|LkWzOlU2KM7:TR?= NF7GRohUSU6JRWK=
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NCI-H1975 MX7DfZRwfG:6aXPpeJkh[XO|YYm= MWW0PEBp M16ybGN6fG:2b4jpZ4l1gSCjZ3HpcpN1KGi3bXHuJG5EUS2KMUm3OUBk\WyuczDh[pRmeiB2ODDodpMh[nliTWTUJIF{e2G7LDDJR|UxRTBwM{myJO69VQ>? M1vZdFIxOTVyNEC0
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PC3 NX\1R|BQS3m2b4TvfIlkcXS7IHHzd4F6 MojnO|IhcA>? M{GzSWN6fG:2b4jpZ4l1gSCjZ3HpcpN1KGi3bXHuJHBEOyClZXzsd{Bi\nSncjC3NkBpenNiYomgUXRVKGG|c3H5MEBKSzVyPUSuNUDPxE1? NY\0bXQxOjBzOEC5NFU>
HeLa NFHBRmdEgXSxdH;4bYNqfHliYYPzZZk> NG\NZVc4OiCq NIXScYdEgXSxdH;4bYNqfHliYXfhbY5{fCCqdX3hckBJ\UyjIHPlcIx{KGGodHXyJFczKGi{czDifUBOXFRiYYPzZZktKEmFNUC9Ok45OSEQvF2= NIPDdlIzODF6MEmwOS=>
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NSCLC NUTCO4p6S3m2b4TvfIlkcXS7IHHzd4F6 M1rF[VczKGh? NInuRWtEgXSxdH;4bYNqfHliYXfhbY5{fCCqdX3hckBPW0OOQzDj[YxteyxiSVO1NF0xNjVibl2= NGO3eoQzODF6MEmwOS=>
Assay
Methods Test Index PMID
Growth inhibition assay Cell apoptosis ; Cell viability 25537503 26515464
Western blot CIP2A / p-AKT / AKT / PARP ; p-EGFR / p-IGF1R / p-ERK / p-AKT 25537503 26052929
Immunofluorescence p65 ; vacuoles 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 (from reference)

Kinase Assay:[1]
  • 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]
  • 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]
  • Animal Models: SCID mice harbouring ARK2 xenografts
  • Dosages: 25 mg/kg
  • Administration: p.o.
  • (Only for Reference)

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

Molecular Weight 717.18
Formula

C24H25ClFN5O3.2C4H4O4

CAS No. 850140-73-7
Storage 3 years -20°C powder
2 years -80°C in solvent
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

In vivo Formulation Calculator (Clear solution)

Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)

mg/kg g μL

Step 2: Enter the in vivo formulation (This is only the calculator, not formulation. Please contact us first if there is no in vivo formulation at the solubility Section.)

% DMSO % % Tween 80 % ddH2O
%DMSO %

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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.

Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.

Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such
as vortex, ultrasound or hot water bath can be used to aid dissolving.

Molarity Calculator

Mass Concentration Volume Molecular Weight

Clinical Trial Information

NCT Number Recruitment Interventions Conditions Sponsor/Collaborators Start Date Phases
NCT04795245 Active not recruiting Drug: afatinib Non-squamous Non-Small Cell Lung Cancer Boehringer Ingelheim August 26 2021 --
NCT04497584 Recruiting Drug: Afatinib + Prednisone Advanced Squamous Non Small Cell Lung Cancer University of Texas Southwestern Medical Center August 4 2021 Phase 2
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

(data from https://clinicaltrials.gov, updated on 2022-01-17)

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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