BAY 43-9006 inhibition of oncogenic RET mutants
BAY 43-9006 inhibition of oncogenic RET mutants
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DOI：
10.1093/jnci/djj069
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发表时间：
2006-03-01
影响因子：
10.3
通讯作者：
Santoro, M
中科院分区：
文献类型：
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作者：
Carlomagno, F;Anaganti, S;Santoro, M
背景。甲状腺髓样癌和乳头状癌通常与 RET 酪氨酸激酶的致癌激活有关。我们评估了联芳基脲 BAY 43-9006（已知可抑制其他几种酪氨酸激酶）是否会阻断 RET 激酶功能和致癌活性。方法：我们通过使用免疫复合物激酶测定和磷酸特异性抗体的免疫印迹，在体外和致癌 RET 转染的 NIH3T3 成纤维细胞中的细胞 RET 信号传导中检测了 BAY 43-9006 抗致癌 RET 的活性。使用生长曲线和流式细胞术测定了 BAY 43-9006 对含有自发致癌 RET 等位基因的人 TPC1 和 TT 甲状腺癌细胞增殖的影响，以及对用致癌 RET 突变体（包括对其他化疗药物耐药的突变体）转化的 RAT1 成纤维细胞增殖的影响。测量了口服 BAY 43-9006 或载体处理的无胸腺小鼠中 TT 细胞衍生的异种移植肿瘤的生长。所有统计检验都是双面的。结果：BAY 43-9006 在 NIH3T3 细胞中以 50 nM 或更低的半数最大抑制浓度 (IC(50)s) 抑制致癌 RET 激酶活性。它还抑制了由致癌 RET 转化的 NIH3T3 和 RATI 成纤维细胞以及含有自发致癌 RET 等位基因的甲状腺癌细胞的生长。此外，BAY 43-9006 抑制携带 RET V804L（IC50 = 110 nM，95% 置信区间 [CI] = 88 至 133 nM）或 RET V804M（IC50 = 147 nM，95% CI = 123 nM 至 170 nM）的细胞的生长，这两种突变体均对苯胺喹唑啉具有抗性，并且吡唑并嘧啶类。 BAY 43-9006（60 mg/kg/天）口服治疗 3 周后，TT 细胞异种移植物（n = 7）的体积从 72.5 减少至 44 mm(3)（差异 = 28.5 mm(3)，95% CI = 7 mm(3) 至 50 mm(3)），而在载体治疗的小鼠（it = 7）中，平均肿瘤体积增加至第408章 mm(3)（差异 = 320 mm(3)，95% CI = 180 mm(3) 至 460 mm(3)；未治疗与治疗比较，P=.02）。这种抑制与 RET 磷酸化的减少平行。结论：BAY 43-9006 是 RET 激酶的强效抑制剂。它作为 RET 阳性甲状腺肿瘤（包括表达 V804 突变的甲状腺肿瘤）治疗工具的潜力值得研究。
Background. Medullary and papillary thyroid carcinomas are often associated with oncogenic activation of the RET tyrosine kinase. We evaluated whether the biaryl urea BAY 43-9006, which is known to inhibit several other tyrosine kinases, blocks RET kinase function and oncogenic activity. Methods: We examined BAY 43-9006 activity against oncogenic RET in vitro and in cellular RET signaling in oncogenic RET-transfected NIH3T3 fibroblasts by using immunocomplex kinase assays and immunoblotting with phospho-specific antibodies. The effects of BAY 43-9006 on proliferation of human TPC1 and TT thyroid carcinoma cells, which harbor spontaneous oncogenic RET alleles, and on RAT1 fibroblasts transformed with oncogenic RET mutants, including mutants that are resistant to other chemotherapeutic agents, were determined using growth curves and flow cytometry. Growth of TT cell-derived xenograft tumors in athymic mice treated orally with BAY 43-9006 or with vehicle was measured. All statistical tests were two-sided. Results: BAY 43-9006 inhibited oncogenic RET kinase activity at half-maximal inhibitory concentrations (IC(50)s) of 50 nM or less in NIH3T3 cells. It also arrested the growth of NIH3T3 and RATI fibroblasts transformed by oncogenic RET and of thyroid carcinoma cells that harbor spontaneous oncogenic RET alleles. Moreover, BAY 43-9006 inhibited the growth of cells carrying RET V804L (IC50 = 110 nM, 95% confidence interval [CI] = 88 to 133 nM) or RET V804M (IC50 = 147 nM, 95% CI = 123 nM to 170 nM), both mutants that are resistant to anilinoquinazolines and pyrazolopyrimidines. After 3 weeks of oral treatment with BAY 43-9006 (60 mg/kg/day), the volume of TT cell xenografts (n = 7) was reduced from 72.5 to 44 mm(3) (difference = 28.5 mm(3), 95% CI = 7 mm(3) to 50 mm(3)), whereas in vehicle-treated mice (it = 7), mean tumor volume increased to 408 mm(3) (difference = 320 mm(3), 95% CI = 180 mm(3) to 460 mm(3); untreated versus treated, P=.02). This inhibition paralleled a decrease in RET phosphorylation. Conclusions: BAY 43-9006 is a powerful inhibitor of the RET kinase. Its potential as a therapeutic tool for RET-positive thyroid tumors, including those expressing V804 mutations merits study.