Afatinib (BIBW2992)

製品コード:S1011

For research use only.

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

Afatinib (BIBW2992)化学構造

CAS No. 850140-72-6

サイズ 価格(税別)
10mM (1mL in DMSO) JPY 41800
JPY 21900
JPY 96600
JPY 212800
JPY 361600

代表番号: 03-5615-9297|電子メール:[email protected]
よく尋ねられる質問

文献中Selleckの製品使用例(336)

製品安全説明書

EGFR阻害剤の選択性比較

相関EGFR製品

生物活性

製品説明 Afatinib (BIBW2992) inhibits EGFR/ErbB irreversibly in vitro with IC50 of 0.5, 0.4, 10, 14, 1 nM for EGFRwt, EGFR L858R , EGFR L858R/T790M ErbB2 (HER2) and ErbB4 (HER4), respectively. Afatinib induces autophagy.
ターゲット
EGFR (L858R) [1]
(Cell-free assay)
EGFR (wt) [1]
(Cell-free assay)
ErbB4 [2]
(Cell-free assay)
EGFR (L858R/T790M) [1]
(Cell-free assay)
HER2 [1]
(Cell-free assay)
0.4 nM 0.5 nM 1 nM 10 nM 14 nM
体外試験

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]

細胞データ
Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
HDLM-2 NHHFe2xIem:5dHigTY5pcWKrdHnvckBCe3OjeR?= M3HjOmlEPTB;MD6wNFU4OiEQvF2= MUO8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:5d4eu[YJqNmGlLoXrM4Np\W2kbD;jc41xd3WwZG;y[ZBwenShY3Hy[E9EUEWPQlyxNVc{PjV3Lze+V2FPT0WUPD;hQi=>
KASUMI-1 M3rBdWdzd3e2aDDJcohq[mm2aX;uJGF{e2G7 M2Dm[2lEPTB;MD6wNFkyOSEQvF2= MkHoQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xd4f3MoVjcS6jYz71b{9kcGWvYnyvZ49ueG:3bnTfdoVxd3K2X3PhdoQwS0iHTVLMNVE4OzZ3NT:nQnNCVkeHUkyvZV4>
HCC2218 MYnHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? NEP2T|FKSzVyPUCuNFA6QDdizszN Ml;JQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xd4f3MoVjcS6jYz71b{9kcGWvYnyvZ49ueG:3bnTfdoVxd3K2X3PhdoQwS0iHTVLMNVE4OzZ3NT:nQnNCVkeHUkyvZV4>
KARPAS-45 NX:wNHJVT3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm= MYXJR|UxRTBwMEG2O|Eh|ryP MlHXQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xd4f3MoVjcS6jYz71b{9kcGWvYnyvZ49ueG:3bnTfdoVxd3K2X3PhdoQwS0iHTVLMNVE4OzZ3NT:nQnNCVkeHUkyvZV4>
HSC-4 NXTBSWJbT3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm= Mo[1TWM2OD1yLkCxO|Q5KM7:TR?= NFPLeGY9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;3e5cv\WKrLnHjMpVsN2OqZX3icE9kd22yb4Xu[H9z\XCxcoTfZ4Fz\C:FSFXNRmwyOTd|NkW1M{c,W0GQR1XSQE9iRg>?
CHL-1 NUTncIJ4T3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm= MVXJR|UxRTBwMEG4N|kh|ryP NFjxTo49[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;3e5cv\WKrLnHjMpVsN2OqZX3icE9kd22yb4Xu[H9z\XCxcoTfZ4Fz\C:FSFXNRmwyOTd|NkW1M{c,W0GQR1XSQE9iRg>?
KY821 M1jYZWdzd3e2aDDJcohq[mm2aX;uJGF{e2G7 MWLJR|UxRTBwMEKwOkDPxE1? MoD5QIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xd4f3MoVjcS6jYz71b{9kcGWvYnyvZ49ueG:3bnTfdoVxd3K2X3PhdoQwS0iHTVLMNVE4OzZ3NT:nQnNCVkeHUkyvZV4>
NCI-N87 NWPsdJJkT3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm= NH\uSlFKSzVyPUCuNFI4PzNizszN MUG8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:5d4eu[YJqNmGlLoXrM4Np\W2kbD;jc41xd3WwZG;y[ZBwenShY3Hy[E9EUEWPQlyxNVc{PjV3Lze+V2FPT0WUPD;hQi=>
DOK MnnlS5Jwf3SqIFnubIljcXSrb36gRZN{[Xl? MmnHTWM2OD1yLkCzNVk1KM7:TR?= MlKwQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xd4f3MoVjcS6jYz71b{9kcGWvYnyvZ49ueG:3bnTfdoVxd3K2X3PhdoQwS0iHTVLMNVE4OzZ3NT:nQnNCVkeHUkyvZV4>
DSH1 NWXZbmR[T3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm= NV3ycJp{UUN3ME2wMlA{OzF2IN88US=> M1\mOVxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4f3e{5m[mlwYXOueYsw[2inbXLsM4NwdXCxdX7kY5JmeG:{dG;jZZJlN0OKRV3CUFEyPzN4NUWvK|5USU6JRWK8M4E,
MDA-MB-175-VII Mm\pS5Jwf3SqIFnubIljcXSrb36gRZN{[Xl? NIjx[IFKSzVyPUCuNFM1OTZizszN MoS0QIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xd4f3MoVjcS6jYz71b{9kcGWvYnyvZ49ueG:3bnTfdoVxd3K2X3PhdoQwS0iHTVLMNVE4OzZ3NT:nQnNCVkeHUkyvZV4>
NCI-H2170 NXfJ[FM3T3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm= NXnNOGJQUUN3ME2wMlA{PzJ6IN88US=> Ml;pQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xd4f3MoVjcS6jYz71b{9kcGWvYnyvZ49ueG:3bnTfdoVxd3K2X3PhdoQwS0iHTVLMNVE4OzZ3NT:nQnNCVkeHUkyvZV4>
BT-474 M3rxXmdzd3e2aDDJcohq[mm2aX;uJGF{e2G7 M361dmlEPTB;MD6wOFk5PiEQvF2= NIP5XIY9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;3e5cv\WKrLnHjMpVsN2OqZX3icE9kd22yb4Xu[H9z\XCxcoTfZ4Fz\C:FSFXNRmwyOTd|NkW1M{c,W0GQR1XSQE9iRg>?
EFO-27 M4Tye2dzd3e2aDDJcohq[mm2aX;uJGF{e2G7 M4DDXGlEPTB;MD6wOVE5OiEQvF2= MWq8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:5d4eu[YJqNmGlLoXrM4Np\W2kbD;jc41xd3WwZG;y[ZBwenShY3Hy[E9EUEWPQlyxNVc{PjV3Lze+V2FPT0WUPD;hQi=>
NCI-H720 NHLPcFJIem:5dHigTY5pcWKrdHnvckBCe3OjeR?= MWnJR|UxRTBwMEWzNlUh|ryP NWH6e3M{RGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:ve5d4NmWkaT7hZ{52cy:laHXtZoww[2:vcH;1coRgemWyb4L0Y4NiemRxQ1jFUWJNOTF5M{[1OU8oRlODTlfFVlww[T5?
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KYSE-450 M2jBb2dzd3e2aDDJcohq[mm2aX;uJGF{e2G7 NYjvZm57UUN3ME2wMlA2QTd6IN88US=> MofLQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xd4f3MoVjcS6jYz71b{9kcGWvYnyvZ49ueG:3bnTfdoVxd3K2X3PhdoQwS0iHTVLMNVE4OzZ3NT:nQnNCVkeHUkyvZV4>
BPH-1 NHnjTHJIem:5dHigTY5pcWKrdHnvckBCe3OjeR?= NH;1NXRKSzVyPUCuNFU6QDhizszN NYK1UGhvRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:ve5d4NmWkaT7hZ{52cy:laHXtZoww[2:vcH;1coRgemWyb4L0Y4NiemRxQ1jFUWJNOTF5M{[1OU8oRlODTlfFVlww[T5?
NCI-H292 M2XUSWdzd3e2aDDJcohq[mm2aX;uJGF{e2G7 NHj1VHRKSzVyPUCuNFYxQSEQvF2= MUC8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:5d4eu[YJqNmGlLoXrM4Np\W2kbD;jc41xd3WwZG;y[ZBwenShY3Hy[E9EUEWPQlyxNVc{PjV3Lze+V2FPT0WUPD;hQi=>
H9 NGPRVIZIem:5dHigTY5pcWKrdHnvckBCe3OjeR?= M2n0WWlEPTB;MD6wO|E{OiEQvF2= M122T|xiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4f3e{5m[mlwYXOueYsw[2inbXLsM4NwdXCxdX7kY5JmeG:{dG;jZZJlN0OKRV3CUFEyPzN4NUWvK|5USU6JRWK8M4E,
NCI-H526 MYPHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? NXXJOpdwUUN3ME2wMlA4QDJ|IN88US=> MoCzQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xd4f3MoVjcS6jYz71b{9kcGWvYnyvZ49ueG:3bnTfdoVxd3K2X3PhdoQwS0iHTVLMNVE4OzZ3NT:nQnNCVkeHUkyvZV4>
NCI-H1623 MVzHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? M17PemlEPTB;MD6wPFAzPSEQvF2= NF2xflg9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;3e5cv\WKrLnHjMpVsN2OqZX3icE9kd22yb4Xu[H9z\XCxcoTfZ4Fz\C:FSFXNRmwyOTd|NkW1M{c,W0GQR1XSQE9iRg>?
KYSE-140 NFf3OFZIem:5dHigTY5pcWKrdHnvckBCe3OjeR?= MVfJR|UxRTBwMEizN{DPxE1? M4LFN|xiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4f3e{5m[mlwYXOueYsw[2inbXLsM4NwdXCxdX7kY5JmeG:{dG;jZZJlN0OKRV3CUFEyPzN4NUWvK|5USU6JRWK8M4E,
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ZR-75-30 M4jCVWdzd3e2aDDJcohq[mm2aX;uJGF{e2G7 MUPJR|UxRTBwMEm1OFUh|ryP MYC8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:5d4eu[YJqNmGlLoXrM4Np\W2kbD;jc41xd3WwZG;y[ZBwenShY3Hy[E9EUEWPQlyxNVc{PjV3Lze+V2FPT0WUPD;hQi=>
NCI-H1770 MlzDS5Jwf3SqIFnubIljcXSrb36gRZN{[Xl? NVLoTmlSUUN3ME2wMlExOjhizszN MWG8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:5d4eu[YJqNmGlLoXrM4Np\W2kbD;jc41xd3WwZG;y[ZBwenShY3Hy[E9EUEWPQlyxNVc{PjV3Lze+V2FPT0WUPD;hQi=>
LB2241-RCC MVXHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? MYLJR|UxRTBwMUC4PFQh|ryP NHi4XZE9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;3e5cv\WKrLnHjMpVsN2OqZX3icE9kd22yb4Xu[H9z\XCxcoTfZ4Fz\C:FSFXNRmwyOTd|NkW1M{c,W0GQR1XSQE9iRg>?
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アッセイ
Methods Test Index PMID
Western blot CIP2A / p-AKT / AKT / PARP ; p-EGFR / p-IGF1R / p-ERK / p-AKT ; p-VEGFR2 / VEGFR2 / JAG1 / DLL4 / NICD1 25537503 26052929 28533486
Growth inhibition assay Cell apoptosis ; Cell viability 25537503 26515464
Immunofluorescence p65 ; vacuoles 26317651 29713246
体内試験 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]

お薦めの試験操作(参考用のみ)

キナーゼ試験:

[1]

  • 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.

細胞試験:

[1]

  • 細胞株:NSCLC cells
  • 濃度: 0-10 μM
  • 反応時間:1 hour
  • 実験の流れ:

    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.

動物試験:

[1]

  • 動物モデル:Athymic NMRI-nu/nu female mice
  • 投薬量:20 mg/kg
  • 投与方法:Oral administration

溶解度 (25°C)

体外

体内

左から(NMPから)右の順に溶剤を製品に加えます(文献ではなく、Selleckの実験によるデータ):
2% DMSO+30% PEG 300+5% Tween 80+ddH2O
混合させたのち直ちに使用することを推奨します。

10mg/mL

化学情報

分子量 485.94
化学式

C24H25ClFN5O3

CAS No. 850140-72-6
Storage 3年 -20°C
2年 -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

投与溶液組成計算機(クリア溶液)

ステップ1:実験データを入力してください。(実験操作によるロスを考慮し、動物数を1匹分多くして計算・調製することを推奨します)

mg/kg g μL

ステップ2:投与溶媒の組成を入力してください。(ロット毎に適した溶解組成が異なる場合があります。詳細については弊社までお問い合わせください)

% DMSO % % Tween 80 % ddH2O
%DMSO %

計算結果:

投与溶媒濃度: mg/ml;

DMSOストック溶液調製方法: mg 試薬を μL DMSOに溶解する(濃度 mg/mL, 注:濃度が当該ロットのDMSO溶解度を超える場合はご連絡ください。 )

投与溶媒調製方法:Take μL DMSOストック溶液に μL PEG300,を加え、完全溶解後μL Tween 80,を加えて完全溶解させた後 μL ddH2O,を加え完全に溶解させます。

投与溶媒調製方法:μL DMSOストック溶液に μL Corn oil,を加え、完全溶解。

注意:1.ストック溶液に沈殿、混濁などがないことをご確認ください;
2.順番通りに溶剤を加えてください。次のステップに進む前に溶液に沈殿、混濁などがないことを確認してから加えてください。ボルテックス、ソニケーション、水浴加熱など物理的な方法で溶解を早めることは可能です。

便利ツール

質量 濃度 体積 分子量

臨床試験

NCT Number Recruitment Interventions Conditions Sponsor/Collaborators Start Date Phases
NCT05298176 Recruiting Drug: Afatinib Osimertinib Carboplatin and Pemetrexed Non-Small Cell Lung Cancer Amsterdam UMC location VUmc January 4 2022 Phase 2
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 Active not recruiting Drug: Afatinib Non-squamous Non-Small Cell Lung Cancer Boehringer Ingelheim May 20 2020 --

(data from https://clinicaltrials.gov, updated on 2022-08-01)

技術サポート

ストックの作り方、阻害剤の保管方法、細胞実験や動物実験の際に注意すべき点など、製品を取扱う時に問い合わせが多かった質問に対しては取扱説明書でお答えしています。

Handling Instructions

他に質問がある場合は、お気軽にお問い合わせください。

* 必須

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よくある質問(FAQ)

質問1:
What’s the recommendation about reconstitution of the compound for in vivo animal study?

回答
For in vivo experiment of afatinib, we recommend 0.5% methylcellulose/0.2% Tween 80 as vehicle.

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