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分别。拉帕替尼治疗导致EGFR和过表达ErbB-2的肿瘤细胞系的IC50值≤0.16μM[2]。",{"key":254,"value":255},"体内研究","拉帕替尼(GW2016)有效抑制BT474和HN5人肿瘤异种移植物的生长。两种模型的剂量反应性抑制发生在每天两次口服30和100mg\u002Fkg拉帕替尼的荷瘤小鼠的治疗中。在100mg\u002Fkg剂量下观察到肿瘤生长的完全抑制。在该剂量下,在21天治疗过程中,治疗动物体重减轻\u003C10％。拉帕替尼治疗以剂量敏感的方式抑制HN5和BT474细胞的肿瘤异种移植物生长,口服30和100mg\u002Fkg,每日两次,在较高剂量下完全抑制肿瘤生长[1]。拉帕替尼(100 mg\u002Fkg \u002F天,口服强饲法)在大鼠心脏组织中诱导严重的氧化损伤[3]。",{"key":257,"value":258},"激酶实验","从杆状病毒表达系统中纯化EGFR，ErbB-2和ErbB4的细胞内激酶结构域。 EGFR，ErbB-2和ErbB-4反应在96孔聚苯乙烯圆底板中进行，终体积为45μL。反应混合物含有50mM 4-吗啉丙磺酸（pH 7.5），2mM MnCl2,10μMATP，1μCi[γ-33P] ATP \u002F反应，50μM肽A [生物素 - （氨基己酸）-EEEEYFELVAKKK-CONH2 ]，1mM二硫苏糖醇和1μL含有拉帕替尼连续稀释液的DMSO，起始浓度为10μM。通过添加指定的纯化的1型受体细胞内结构域来引发反应。加入的酶量为1pmol \u002F反应（20nM）。通过在水中加入45μL的0.5％磷酸，在23℃下10分钟后终止反应。将终止的反应混合物（75μL）转移至磷酸纤维素滤板。将板过滤并用200μL的0.5％磷酸洗涤三次。向每个孔中加入闪烁混合物（50μL），并通过在Packard Topcount中计数来定量测定[1]。",{"key":260,"value":261},"细胞实验","将细胞以下列密度接种于培养基中的96孔板中：HFF和HN5,1000细胞\u002F孔和BT474,5000细胞\u002F孔。 24小时后，将细胞暴露于载体（0.3％DMSO）或拉帕替尼（1nM，10nM，100nM，1μM，10μM和100μM）。 72小时后从细胞中除去拉帕替尼，并用含有10％FBS和50μg\u002F mL庆大霉素（HFF和HN5）的DMEM或含有10％FBS和50μg\u002F mL庆大霉素（BT474）的RPMI替换。亚甲蓝染色在总时间为16天的时间点进行[1]。",{"key":263,"value":264},"动物实验","小鼠[1] CD-1裸雌性小鼠用于HN5人肿瘤异种移植物，其通过注射PBS：Matrigel（1：1）中的细胞悬浮液而起始。 CB-17 SCID雌性小鼠用于BT474人肿瘤异种移植物，其通过从已建立的肿瘤植入肿瘤片段（20-100mg）而起始。通过皮下注射右侧腹部植入肿瘤细胞和碎片。用卡尺测量sc肿瘤，并且每周称重两次小鼠。使用该公式从肿瘤体积估计肿瘤重量：长度×宽度2\u002F2 =肿瘤体积（mm 3）。当肿瘤可触及，直径3-5毫米时开始治疗。拉帕替尼（30和100mg \u002F kg）在磺基丁基醚-β-环糊精10％水溶液（CD10）的载体中每天两次给药，持续21天。将大鼠[3] Wistar大鼠（12周龄白化雄性）随机分为三组：对照（C，n = 8），曲妥珠单抗（T，n = 8）和拉帕替尼（L，n = 8）治疗。对照动物未经治疗，但T组和拉帕替尼组中的其他动物与化疗药物一起给药。曲妥珠单抗在研究的第一天通过腹膜内注射以10mg \u002F kg \u002F天的剂量递送一次。拉帕替尼每天以100mg \u002F kg \u002F天的剂量通过口服强饲法连续给药7天。所选剂量与诊所使用的剂量相当。在第8天，通过单次腹膜内注射氯胺酮和甲苯噻嗪（分别为50和5mg \u002F kg）诱导麻醉。收集血液样品并取出心脏用于生化分析。",{"key":266,"value":267},"参考文献","[1].\nRusnak DW, et al. The effects of the novel, reversible epidermal growth factor receptor\u002FErbB-2 tyrosine kinase inhibitor, GW2016, on the growth of human normal and tumor-derived cell lines in vitro and in vivo. Mol Cancer Ther. 2001 Dec;1(2):85-94\n[2].\nO'Neill F, et al. Gene expression changes as markers of early lapatinib response in a panel of breast cancer cell lines. Mol Cancer. 2012 Jun 18;11(1):41.\n[3].\nEryilmaz U, et al. S100A1 as a Potential Diagnostic Biomarker for Assessing Cardiotoxicity and Implications for the Chemotherapy of Certain Cancers. PLoS One. 2015 Dec 18;10(12):e0145418.\n[4].\nNi J, et al. Combination inhibition of PI3K and mTORC1 yields durable remissions in mice bearing orthotopic patient-derived xenografts of HER2-positive breast cancer brain metastases. Nat Med. 2016 Jul;22(7):723-6.\n[5].\nOncogene. 2016 Jun 9;35(23):2961-70. doi: 10.1038\u002Fonc.2015.377. 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