Afatinib (BIBW2992) Dimaleate

For research use only. Not for use in humans.

目录号:S7810 中文名称:马来酸阿法替尼

Afatinib (BIBW2992) Dimaleate Chemical Structure

CAS No. 850140-73-7

Afatinib (BIBW2992) Dimaleate 不可逆地抑制 EGFR/HER2,包括 EGFR(wt)EGFR(L858R),EGFR(L858R/T790M) 和 HER2,IC50 分别为0.5 nM,0.4 nM,10 nM 和 14 nM;活性比对Gefitinib抵抗型L858R-T790M EGFR突变体高100倍。Afatinib (BIBW2992) Dimaleate 可诱导自噬。

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客户使用Selleck生产的Afatinib (BIBW2992) Dimaleate发表文献131篇:

产品安全说明书

EGFR抑制剂选择性比较

生物活性

产品描述 Afatinib (BIBW2992) Dimaleate 不可逆地抑制 EGFR/HER2,包括 EGFR(wt)EGFR(L858R),EGFR(L858R/T790M) 和 HER2,IC50 分别为0.5 nM,0.4 nM,10 nM 和 14 nM;活性比对Gefitinib抵抗型L858R-T790M EGFR突变体高100倍。Afatinib (BIBW2992) Dimaleate 可诱导自噬。
靶点
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
体外研究

在表达野生型(H1666)或L858R/T790M (NCI-H1975) EGFR的肺癌细胞系中,Afatinib比erlotinib、gefitinib和lapatinib能够更有效的抑制细胞生长。Afatinib同样对表达HER2 776insV (NCI-H1781) 或EGFR E746_A750del (HCC827)的NSCLC细胞系有效,但对表达野生型EGFR 和HER2的A549细胞无效。[1] Afatinib可增强topotecan和mitoxantrone对SP细胞的细胞毒性,并增强topotecan和mitoxantrone对SP细胞的诱导凋亡作用。[1]

Cell Data
Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
HCC827 cell NGKzcohRem:uaX\ldoF1cW:wIHHzd4F6 MWfBcpRqeHKxbHnm[ZJifGm4ZTDhZ5Rqfmm2eTDh[4FqdnO2IHj1cYFvKEiFQ{iyO{Bk\WyuczDh[pRmeiB7NjDodpMh[nliTWTTJIF{e2G7LDDJR|UxRThibl2= NETR[HMzODV3MEKxNi=>
NCI-H1563 MmqzS5Jwf3SqIHnubIljcXSrb36gZZN{[Xl? NGq1ZlJKdmirYnn0bY9vKG:oIHj1cYFvKE6FST3INVU3OyClZXzsJIdzd3e2aDDpckBiKGOnbHygeoli[mmuaYT5JIF{e2G7LjDJR|UxRTlwN{[wN|Eh|ryP M3jBOXNCVkeHUh?=
NCI-H1573 Mk\VS5Jwf3SqIHnubIljcXSrb36gZZN{[Xl? NF\6RVhKdmirYnn0bY9vKG:oIHj1cYFvKE6FST3INVU4OyClZXzsJIdzd3e2aDDpckBiKGOnbHygeoli[mmuaYT5JIF{e2G7LjDJR|UxRTBwN{O2N{DPxE1? MorhV2FPT0WU
NCI-H1581 MWnHdo94fGhiaX7obYJqfGmxbjDhd5NigQ>? Mnu4TY5pcWKrdHnvckBw\iCqdX3hckBPS0lvSEG1PFEh[2WubDDndo94fGhiaX6gZUBk\WyuII\pZYJqdGm2eTDhd5NigS5iSVO1NF0zQS5|MUS1JO69VQ>? NGO2bnBUSU6JRWK=
NCI-H1648 M3W2Rmdzd3e2aDDpcohq[mm2aX;uJIF{e2G7 MmDZTY5pcWKrdHnvckBw\iCqdX3hckBPS0lvSEG2OFgh[2WubDDndo94fGhiaX6gZUBk\WyuII\pZYJqdGm2eTDhd5NigS5iSVO1NF0xNjR2MUiyJO69VQ>? NIn0WYpUSU6JRWK=
NCI-H1793 MlLtS5Jwf3SqIHnubIljcXSrb36gZZN{[Xl? NFz3dYhKdmirYnn0bY9vKG:oIHj1cYFvKE6FST3INVc6OyClZXzsJIdzd3e2aDDpckBiKGOnbHygeoli[mmuaYT5JIF{e2G7LjDJR|UxRTF2LkKwPFch|ryP NV7vN21pW0GQR1XS
NCI-H1838 NX3sZlY4T3Kxd4ToJIlvcGmkaYTpc44h[XO|YYm= NXzvPIgxUW6qaXLpeIlwdiCxZjDoeY1idiCQQ1mtTFE5OzhiY3XscEBoem:5dHigbY4h[SClZXzsJJZq[WKrbHn0fUBie3OjeT6gTWM2OD1yLk[wNlk6KM7:TR?= M{DTbnNCVkeHUh?=
NCI-H1770 M3LneGdzd3e2aDDpcohq[mm2aX;uJIF{e2G7 M324eWlvcGmkaYTpc44hd2ZiaIXtZY4hVkOLLVixO|cxKGOnbHyg[5Jwf3SqIHnuJIEh[2WubDD2bYFjcWyrdImgZZN{[XlwIFnDOVA:OC5zMEK4JO69VQ>? M2\vfXNCVkeHUh?=
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NCI-H1975 NHjzdlRIem:5dHigbY5pcWKrdHnvckBie3OjeR?= NYLlUJlHUW6qaXLpeIlwdiCxZjDoeY1idiCQQ1mtTFE6PzViY3XscEBoem:5dHigbY4h[SClZXzsJJZq[WKrbHn0fUBie3OjeT6gTWM2OD1yLk[xO|I5KM7:TR?= MWfTRW5ITVJ?
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NCI-H441 NFfEblJIem:5dHigbY5pcWKrdHnvckBie3OjeR?= NXjJVpU2UW6qaXLpeIlwdiCxZjDoeY1idiCQQ1mtTFQ1OSClZXzsJIdzd3e2aDDpckBiKGOnbHygeoli[mmuaYT5JIF{e2G7LjDJR|UxRTNwNUm4OFEh|ryP NUSxeHA5W0GQR1XS
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NCI-H23 NIfZVlBIem:5dHigbY5pcWKrdHnvckBie3OjeR?= MUfJcohq[mm2aX;uJI9nKGi3bXHuJG5EUS2KMkOgZ4VtdCCpcn;3eIghcW5iYTDj[YxtKH[rYXLpcIl1gSCjc4PhfU4hUUN3ME2zO{41OTl4IN88US=> NFfmWJZUSU6JRWK=
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... Click to View More Cell Line Experimental Data

Assay
Methods Test Index PMID
Growth inhibition assay
Cell apoptosis; 

PubMed: 25537503     


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     


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     


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     


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     


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     


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
体内研究 在MDA-MB-453移植瘤模型中,Afatinib(20 mg/kg,口服给药)可显著诱导肿瘤衰退,累积治疗/对照肿瘤体积比(T/C比)为2%,并下调EGFR和AKT磷酸化水平。[1] 在A7、A431、FaDu、UT-SCC-14和UT-SCC-15移植瘤模型中,Afatinib(30 mg/kg,口服给药)可显著延长肿瘤生长时间。[3] 在HER2扩增移植瘤模型中,Afatinib(30 mg/kg,口服给药)可显著抑制肿瘤生长,并显著延长生存期。[4] 在HER2阳性胃癌NCI-N87移植瘤模型中,Afatinib(25 mg/kg)口服给药4天可显著减小肿瘤体积,给药21天几乎完全治愈肿瘤。[5]

推荐的实验操作(此推荐来自于公开的文献所以Selleck并不保证其有效性)

激酶实验:[1]
- 合并

体外激酶实验:

EGFR激酶:每100 μL酶反应体系含10 μl 溶于50% Me2SO的抑制剂,20 μL底物溶液(200Mm HEPES pH 7.4,50 mM乙酸镁,2.5 mg/mL聚(EY),5μg/mlbio-pEY)和20 μL酶。加入50μl 100 μM ATP(溶于10 mM MgCl2)开始酶促反应,室温下反应30分钟,加入50 μl终止液终止反应(250 mM EDTA溶于20 mM pH 7.4 HEPES)。吸取100 μL到链霉亲和素包被微孔中,室温温育60 min后,用200 μL/孔洗涤液(50 mM Tris,0.05% 吐温20)洗涤。每孔加入100 μL 250 ng/mL HRPO标记的anti-PY抗体(Py20H Anti-Ptyr:HRP)。反应60 min后,用200 μL洗涤液洗涤三次。加入100 μL TMB过氧化物酶溶液(A:B= 1:1)。反应10分钟后终止反应,ELISA检测器测定OD450 nm处吸光值。HER2-IC 酶:酶活性在50%Me2SO中,不同浓度的抑制剂存在或不存在下测定。每100μL酶反应体系和EGFR激酶检测体系类似,再加入1000 μM Na3VO4。在10mM醋酸镁中加入50 μL 500 μM ATP溶液开始酶促反应。酶需经过稀释,以含130 mM NaCl、0.05% Triton X-100、1 mM DTT和10% 甘油的20 mM pH 7.4 的HEPES稀释。室温反应30 min,加入50 μL终止液终止。Src激酶的测定:每100μL反应包含10 μL 含抑制剂的50% Me2SO,20 μL酶,20 μL 以1000 μM Na3VO4配置的底物。加入50 μL 1000 μM ATP溶液开始酶促反应。BIRK激酶测定:250 mM pH 7.4 Tris-HCl、10 mM DTT、2.5 mg/mL poly(EY)、5 mg/mL bio-pEY作为底物溶剂,加入由8 mM MnCl2和20 mM 醋酸镁配置的2 mM ATP开始酶促反应。VEGF2和HGFR 激酶实验:室温反应20 min,加入10 μL 5%磷酸终止。用96孔过滤收集板收集沉淀到GF/B过滤器中,经洗涤后,50℃干燥1 h,密封和结合放射性由闪烁计数用TopCount™或Microbeta b counter™检测放射性强度。
细胞实验:[2]
- 合并
  • Cell lines: SP和NSP细胞系
  • Concentrations: 1 μM
  • Incubation Time: 48 h
  • Method: 采用MTT法测定细胞毒性。IC50值为导致50%细胞死亡的药物浓度。用Bliss法拟合剂量应答曲线和计算IC50值。
    (Only for Reference)
动物实验:[4]
- 合并
  • Animal Models: 负荷ARK2移植瘤的SCID小鼠
  • Dosages: 25 mg/kg
  • Administration: 口服给药
    (Only for Reference)

溶解度 (25°C)

体外 DMSO 100 mg/mL (139.43 mM)
Water Insoluble
Ethanol Insoluble
体内 从左到右依次将纯溶剂加入产品,现配现用(数据来自Selleck实验检测而非文献):
5% DMSO+30% PEG 300+ddH2O
28mg/mL

* 溶解度检测是由Selleck技术部门检测的,可能会和文献中提供的溶解度有所差异,这是由于生产工艺和批次不同产生的正常现象。请按照顺序依次加入各个纯溶剂。

化学数据

分子量 717.18
化学式

C24H25ClFN5O3.2C4H4O4

CAS号 850140-73-7
储存条件 粉状
溶于溶剂
别名 N/A

动物体内配方计算器 (澄清溶液)

第一步:请输入基本实验信息(考虑到实验过程中的损耗,建议多配一只动物的药量)
给药剂量 mg/kg 动物平均体重 g 每只动物给药体积 ul 动物数量
第二步:请输入动物体内配方组成(配方适用于不溶于水的药物;不同批次药物配方比例不同,请联系Selleck为您提供正确的澄清溶液配方)
% DMSO % % Tween 80 % ddH2O
计算重置

计算器

摩尔浓度计算器

摩尔浓度计算器

本计算器可帮助您计算出特定溶液中溶质的质量、溶液浓度和体积之间的关系,公式为:

质量 (mg) = 浓度 (mM) x 体积 (mL) x 分子量 (g/mol)

摩尔浓度计算公式

  • 质量
    浓度
    体积
    分子量

*在配置溶液时,请务必参考Selleck产品标签上、SDS / COA(可在Selleck的产品页面获得)批次特异的分子量使用本工具。

稀释计算器

稀释计算器

用本工具协助配置特定浓度的溶液,使用的计算公式为:

开始浓度 x 开始体积 = 最终浓度 x 最终体积

稀释公式

稀释公式一般简略地表示为: C1V1 = C2V2 ( 输入 输出 )

  • C1
    V1
    C2
    V2

在配置溶液时,请务必参考Selleck产品标签上、SDS / COA(可在Selleck的产品页面获得)批次特异的分子量使用本工具。.

连续稀释计算器方程

  • 连续稀释

  • 计算结果

  • C1=C0/X C1: LOG(C1):
    C2=C1/X C2: LOG(C2):
    C3=C2/X C3: LOG(C3):
    C4=C3/X C4: LOG(C4):
    C5=C4/X C5: LOG(C5):
    C6=C5/X C6: LOG(C6):
    C7=C6/X C7: LOG(C7):
    C8=C7/X C8: LOG(C8):
分子量计算器

分子量计算器

通过输入化合物的化学式来计算其分子量:

总分子量:g/mol

注:化学分子式大小写敏感。C10H16N2O2 c10h16n2o2

摩尔浓度计算器

质量 浓度 体积 分子量
计算

临床试验信息

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

技术支持

在订购、运输、储存和使用我们的产品的任何阶段,您遇到的任何问题,均可以通过拨打我们的热线电话400-668-6834,或者技术支持邮箱tech@selleck.cn,直接联系到我们。我们会在24小时内尽快联系您。

操作手册

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Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID