Target Binding Properties and Cellular Activity of Afatinib (BIBW 2992), an Irreversible ErbB Family Blocker
Target Binding Properties and Cellular Activity of Afatinib (BIBW 2992), an Irreversible ErbB Family Blocker
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DOI：
10.1124/jpet.112.197756
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发表时间：
2012-11-01
影响因子：
3.5
通讯作者：
Adolf, Guenther R.
中科院分区：
文献类型：
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作者：
Solca, Flavio;Dahl, Goeran;Adolf, Guenther R.
禽红母细胞病病毒（AEV-H株）原癌基因B受体网络的解除管制被认为是上皮癌的致癌驱动因素。已经开发了几种靶向药物，包括抗体和小分子激酶抑制剂，每种药物都具有不同的ErbB受体相互作用模式。了解这些化合物的确切药理性质对于在临床实践中最佳使用是重要的。阿法替尼[BIBW 2992；N-[4-[（3-氯-4-氟苯基）氨基]-7-[[(3S)-四氢-3-呋喃基]氧]-6-喹唑啉基]4-（二甲氨基）-2-丁烯酰胺]是一种具有atp竞争性的苯胺喹唑啉衍生物，含有活性丙烯酰胺基团。它被设计成共价结合和不可逆地阻断酶活性ErbB受体家族成员。在这里，我们通过x射线晶体学显示了阿法替尼与野生型表皮生长因子受体（EGFR）的共价结合，并通过质谱分析显示了与EGFR、EGFR（L858R/T790M）、人表皮生长因子受体2 （HER2）和ErbB-4的共价相互作用。在浓度低于100 nM时，阿法替尼能有效抑制ErbB-4 （EC50 = 1 nM）的酶活性和多种ErbB受体畸变驱动的癌细胞的增殖。N-[4-[（3-氯-4-氟苯基）氨基]-7-[[(3S)-四氢-3-呋喃基]氧]-6-喹唑啉基]-4-（二甲氨基）-2-丁酰胺（BI 37781）是阿法替尼的类似物，缺乏丙烯酰胺基团，因此不能形成共价键，对EGFR或EGFR（L858R）驱动的细胞具有相似的效力，但对表达EGFR(L858R/T790M) HER2或ErbB-4的细胞活性较低或没有检测到活性。这些结果强调了丙烯酰胺基团的重要性，并表明阿法替尼不同于已批准的ErbB靶向药物，其不可逆地抑制所有ErbB家族成员的激酶活性。它们为该化合物独特的药理学特征提供了机制基础，并解释了一些由于继发性突变或ErbB受体“重编程”而对抗体或激酶抑制剂治疗产生耐药性的患者的临床活性。
Deregulation of the ErbB (proto-oncogene B of the avian erythroblastosis virus AEV-H strain) receptor network is well recognized as an oncogenic driver in epithelial cancers. Several targeted drugs have been developed, including antibodies and small-molecule kinase inhibitors, each of them characterized by distinct patterns of ErbB receptor interactions. Understanding the precise pharmacological properties of these compounds is important for optimal use in clinical practice. Afatinib [BIBW 2992; N-[4-[(3-chloro-4-fluorophenyl) amino]-7-[[(3S)-tetrahydro-3-furanyl]oxy]-6-quinazolinyl]4-(dimethylamino)-2-butenamide] is an ATP-competitive anilinoquinazoline derivative harboring a reactive acrylamide group. It was designed to covalently bind and irreversibly block enzymatically active ErbB receptor family members. Here, we show by X-ray crystallography the covalent binding of afatinib to wild-type epidermal growth factor receptor (EGFR) and by mass spectrometry the covalent interaction with EGFR, EGFR(L858R/T790M), human epidermal growth factor receptor 2 (HER2), and ErbB-4. Afatinib potently inhibits the enymatic activity of ErbB-4 (EC50 = 1 nM) and the proliferation of cancer cell lines driven by multiple ErbB receptor aberrations at concentrations below 100 nM. N-[4-[(3-chloro-4-fluorophenyl)amino]-7-[[(3S)-tetrahydro-3-furanyl]oxy]-6-quinazolinyl]-4-(dimethylamino)-2-butanamide (BI 37781), a close analog of afatinib lacking the acrylamide group and thus incapable of covalent bond formation, had similar potency on cells driven by EGFR or EGFR(L858R), but less or no detectable activity on cells expressing EGFR(L858R/T790M) HER2 or ErbB-4. These results stress the importance of the acrylamide group and show that afatinib differs from approved ErbB targeting agents by irreversibly inhibiting the kinase activity of all ErbB family members. They provide a mechanistic rationale for the distinct pharmacological features of this compound and explain the clinical activity seen in some patients who are resistant to antibody or kinase inhibitor therapy because of secondary mutations or ErbB receptor "reprogramming."