DC120, a Novel and Potent Inhibitor of AKT Kinase, Induces Tumor Cell Apoptosis and Suppresses Tumor Growth
DC120, a Novel and Potent Inhibitor of AKT Kinase, Induces Tumor Cell Apoptosis and Suppresses Tumor Growth
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DC120 是一种新型强效 AKT 激酶抑制剂，可诱导肿瘤细胞凋亡并抑制肿瘤生长
DOI：
10.1124/mol.111.077271
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发表时间：
2012-08-01
影响因子：
3.6
通讯作者：
Zhu, Xiao-Feng
中科院分区：
文献类型：
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作者：
Deng, Rong;Yang, Fen;Zhu, Xiao-Feng
蛋白激酶 B/AKT 激酶是磷脂酰肌醇 3-激酶/AKT 信号通路的核心成分，该信号通路在人类癌症中经常过度激活。我们基于 AKT 的 ATP 结合位点设计并合成了一系列 2-嘧啶基-5-酰胺噻唑化合物，其中最有效的化合物 (S)-N-(1-氨基-3-(2,4-二氯苯基)丙-2-基)-2-(2-(甲基氨基)嘧啶-4-基)噻唑-5-甲酰胺 (DC120) 被鉴定可在体外抑制 AKT 活性，基于荧光共振能量转移的 Z'-LYTE 测定的 EC50 为 153 nM。 DC120的抗肿瘤作用在人CNE2和MDA-MB-453细胞系以及CNE2异种移植模型上进行了测试。结果表明，DC120通过诱导细胞凋亡，明显抑制CNE2和MDA-MB-453细胞的增殖，亚G1和膜联蛋白V阳性细胞增多，细胞核内出现典型的细胞凋亡形态变化，并裂解caspase-3。进一步研究表明，转染组成型激活的AKT1的MDA-MB-453细胞对DC120更敏感，而通过短发夹RNA敲低AKT1表达的CNE2细胞对DC120更耐药。更重要的是，DC120 以剂量依赖性和时间依赖性的方式部分减弱叉头转录因子 (FKHR)、FKHRL1、糖原合成酶激酶 3β 和哺乳动物雷帕霉素靶标的磷酸化水平，并导致癌细胞中外源 FKHR 的核积累增加。此外，20 mg/kg/天的 DC120 抑制 CNE2 异种移植肿瘤生长，治疗组/对照组比率为 38.1%，同时肿瘤样品中末端脱氧核苷酸转移酶 UTP 缺口末端标记阳性细胞增加。此外，DC120诱导反馈环路激活丝裂原激活蛋白激酶途径，并且用丝裂原激活蛋白激酶激酶抑制剂1,4-二氨基-2,3-二氰基-1,4-双(甲硫基)丁二烯(U0126)和DC120治疗可协同诱导癌细胞凋亡。这些数据为开发 DC120 来治疗 AKT 水平升高的癌症提供了验证。
Protein kinase B/AKT kinase is the core component of the phosphatidylinositol 3-kinase/AKT signaling pathway, which is frequently hyperactivated in human cancers. We designed and synthesized a series of 2-pyrimidyl-5-amidothiazole compounds based on the ATP binding site of AKT, and the most potent compound, (S)-N-(1-amino-3-(2,4-dichlorophenyl)propan-2-yl)-2-(2-(methylamino)pyrimidin-4-yl)thiazole-5-carboxamide (DC120), was identified to inhibit AKT activity in vitro with an EC50 of 153 nM by a fluorescence resonance energy transfer-based Z′-LYTE assay. The antitumor effect of DC120 was tested on human CNE2 and MDA-MB-453 cell lines and the CNE2 xenograft model. The results showed that DC120 could obviously inhibit the proliferation of CNE2 and MDA-MB-453 cells via induction of apoptosis, with the evidence of increases in sub-G1 and annexin V-positive cells, characteristic morphologic changes of apoptosis in the nucleus, and cleaved caspase-3. Further study showed that MDA-MB-453 cells transfected with constitutively activated AKT1 were more sensitive to DC120,whereas CNE2 cells with knockdown of AKT1 expression by short hairpin RNA were more resistant to DC120. Of more importance, DC120 partially attenuated the phosphorylation levels of forkhead transcription factor (FKHR), FKHRL1, glycogen synthase kinase 3β, and mammalian target of rapamycin in a dose-dependent and time-dependent fashion and led to an increase in the nuclear accumulation of exogenous FKHR in cancer cells. In addition, DC120 at 20 mg/kg/day inhibited the CNE2 xenograft tumor growth with a treated group/control group ratio of 38.1%, accompanied by increasing terminal deoxynucleotidyl transferasedUTP nick-end labeling-positive cells in the tumor sample. In addition, DC120 induced a feedback loop to activate the mitogen-activated protein kinase pathway and treatment with mitogen-activated protein kinase kinase inhibitor 1,4-diamino-2,3-dicyano-1,4-bis(methylthio)butadiene (U0126) and DC120 synergistically induced cancer cell apoptosis. These data provide validation for the development of DC120 to treat cancers displaying elevated levels of AKT.