BIBW2992 阿法替尼

生物活性

Afatinib (BIBW2992)是一种不可逆EGFR/HER2抑制剂，对EGFR(wt)， EGFR(L858R)， EGFR(L858R/T790M)和HER2均有抑制作用，IC50分别为0.5 nM，0.4 nM，10 nM和14 nM；用于Gefitinib抗性的突变型EGFR (L858R-T790M )时，活性增强100倍。BIBW2992不可逆地抑制EGFR/HER2，在体外作用于野生型EGFR ,L858R突变型EGFR, L858R/T790M 双突变型EGFR，及HER2时IC50分别为0.5, 0.4, 10, 14 nM。在体外实验中，BIBW2992不管作用于野生型EGFR/HER2还是突变型EGFR/HER2都显示出强大的活力，与gefitinib作用于L858R突变型EGFR的活力差不多，但是要比gefitinib 作用于L858R-T790M双突变型EGFR的活力大100倍。在体外，BIBW2992有效抑制所有实验类型细胞的EGFR/HER磷酸化作用。

实验操作 来自于公开的文献，仅供相同实验参考（如实验材料、目的不同，请参考其他文献）

细胞实验
细胞系	NSCLC cells H1666, H3255 and NCI-H1975
方法	Growth inhibition of lung cancer cell lines by BIBW2992 and other EGFR and/or HER2 inhibitors (a) Sensitivity of NSCLC cell lines that express EGFR WT, Exon 19 Del, L858R/ T790M and HER2 INS776V to BIBW2992 and erlotinib. Dose-dependent growth inhibition was determined by MTS assay. Only HCC827 cells are sensitive to erlotinib whereas NCIH1781 and NCI-H1975 cells are also sensitive to BIBW2992. Points, average of two independent experiments, each done in triplicate; bars, s.d. *A549 cells harbor a Kras G12S mutation. See also in Supplementary Table 4. (b) Inhibition of anchorage independent cell proliferation of various lung cancer cell lines treated with BIBW2992, lapatinib, canertinib, gefitinib or erlotinib. The IC50 values were determined as described.
浓度	0~10µM
处理时间	72 h

动物实验
动物模型	A431 cells mouse bearing tumor xenograft model
配制	0.5% methocellulose-0.4% polysorbate-80 (Tween 80)
剂量	20 ml/kg body weight once daily
给药处理	oral gavage

不同实验动物依据体表面积的等效剂量转换表（数据来源于FDA指南）

	小鼠	大鼠	兔	豚鼠	仓鼠	狗
重量 (kg)	0.02	0.15	1.8	0.4	0.08	10
体表面积 (m2)	0.007	0.025	0.15	0.05	0.02	0.5
Km 系数	3	6	12	8	5	20

动物 A (mg/kg) = 动物 B (mg/kg) ×	动物 B的Km系数
	动物 A的Km系数

例如，依据体表面积折算法，将白藜芦醇用于小鼠的剂量22.4 mg/kg 换算成大鼠的剂量，需要将22.4 mg/kg 乘以小鼠的K_m系数（3），再除以大鼠的K_m系数（6），得到白藜芦醇用于大鼠的等效剂量为11.2 mg/kg。

化学数据

储备液配制

以下数据基于产品分子量，对于特殊产品，请参照COA中的储备液配制条件和说明进行操作。

Concentration / Solvent Volume / Mass	1 mg	5 mg	10 mg
1 mM	2.0579 mL	10.2893 mL	20.5787 mL
5 mM	0.4116 mL	2.0579 mL	4.1157 mL
10 mM	0.2058 mL	1.0289 mL	2.0579 mL

使用AbMole产品发表的文献

• 

  J Virol. 2020 Feb 14;94(5):e01791-19.

  Targeting Kaposi's Sarcoma-Associated Herpesvirus ORF21 Tyrosine Kinase and Viral Lytic Reactivation by Tyrosine Kinase Inhibitors Approved for Clinical Use
  BIBW2992购买自AbMole

客户产品验证及相关生物数据

	数据来源	Am J Cancer Res (2015). Figure 3. BIBW2992
	方法	Western Blot
	细胞系	HKESC-2 and EC-1 cell lines
	浓度	0.1 μM
	处理时间	24 h
	实验结果	In HKESC-2 and EC-1 cell lines, afatinib could suppress AKT phosphorylation in a dose and time dependent manner, in which the phosphorylation was almost completely abrogated when the cells were treated with concentrations around their corresponding IC50 (HKESC-2: 0.1 μM, EC-1: 1 μM) for 24 hours (Figure 3).

	数据来源	Am J Cancer Res (2015). Figure 1. BIBW2992
	方法	ELISA
	细胞系	ESCC cell lines
	浓度	0.1 μM
	处理时间	48 and 72 h
	实验结果	All 5 ESCC cell lines were sensitive to afatinib, in which over 95% of growth inhibition was observed after 48 and 72 hours of treatment (Figure 1B).

参考文献

A phase II trial to assess efficacy and safety of afatinib in extensively pretreated patients with HER2-negative metastatic breast cancer.
Schuler et al. Breast Cancer Res Treat. 2012 Aug;134(3):1149-59. PMID: 22763464.

A phase II study of afatinib (BIBW 2992), an irreversible ErbB family blocker, in patients with HER2-positive metastatic breast cancer progressing after trastuzumab.
Lin et al. Breast Cancer Res Treat. 2012 Jun;133(3):1057-65. PMID: 22418700.

Clinical activity of afatinib (BIBW 2992) in patients with lung adenocarcinoma with mutations in the kinase domain of HER2/neu.
De Grève et al. Lung Cancer. 2012 Apr;76(1):123-7. PMID: 22325357.