Evaluation of a novel AKT inhibitor in ovarian cancer

新型 AKT 抑制剂治疗卵巢癌的评价

• 批准号: 7017651
• 负责人: Jiayuh Lin
• 金额: $ 12.78万
• 依托单位: RESEARCH INST NATIONWIDE CHILDREN'S HOSP
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-04-01 至 2006-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7017651
• 关键词: antineoplastics; apoptosis; athymic mouse; biological signal transduction; cisplatin; enzyme activity; enzyme inhibitors; ovary neoplasms; paclitaxel; phosphorylation; protein isoforms; protein kinase; protooncogene; small molecule; technology /technique development; terminal nick end labeling; western blottings

DESCRIPTION (provided by applicant): This research project will focus on the evaluation of a novel AKT-selective small molecule inhibitor in ovarian cancer cells and the effect of this inhibitor on AKT downstream targets. AKT or Protein Kinase B (PKB) is a serine/threonine kinase that is activated in response to growth factor or cytokine. The AKT protein has three isoforms AKT1 (PKBalpha), AKT2 (PKBbeta) and AKT3 (PKBgamma) that have greater than 85% sequence identity and have the same structural organization. The AKT2 proto-oncogene is amplified in ovarian carcinoma. The elevated AKT1 phosphorylation and AKT1 kinase activity have also been detected frequently in ovarian cancer cells and tumors. Ovarian cancer patients with AKT alterations appear to have a poor prognosis. Since AKT may provide a survival signal that protects cells from apoptosis induced by anti-cancer drugs or stresses, development of potent and selective inhibitors targeting AKT is an attractive therapeutic strategy for treating ovarian carcinoma. Through structure-based design and screening, we have identified a potent and selective small molecule inhibitor of AKT (termed API-59). In this proposal, we propose to evaluate the potency, selectivity and molecular mechanism of API-59 in ovarian cancer cells that express elevated AKT activity. The following specific aims will be studied to address this concept: 1. Evaluation of the effect of API-59 on AKT pathway in ovarian cancer cells. 1.1. Synthesis of API-59. 1.2. Evaluate induction of apoptosis by API-59 in ovarian cancer cell lines that express constitutively active AKT. 1.3. Determine the effect of API-59 on AKT kinase activity and AKT downstream targets. 1.4. Evaluate the effect of API-59 on normal cells lacking constitutively active AKT. 2. Examination of the effect of API-59 in combination with cytotoxic agents in ovarian cancer cells. 2.1. Test the effect of API-59 in combination with cisplatin on inducing apoptosis and inhibiting the AKT pathway in ovarian cancer cell lines that express constitutively active AKT. 2.2. Examine the inhibitory effect of API-59 in combination with taxol on inducing apoptosis and inhibiting the AKT pathway in ovarian cancer cells that express constitutively active AKT. 3. Evaluation of the efficacy of API-59 in a nude mouse tumor model in vivo. Determine the efficacy of API-59 treatment to inhibit elevated AKT activity in established mice tumors and retard the growth of established tumors in nude mice.

描述（由申请人提供）：本研究项目将重点评估卵巢癌细胞中新型AKT选择性小分子抑制剂以及该抑制剂对AKT下游靶点的影响。AKT或蛋白激酶B（PKB）是响应于生长因子或细胞因子而被激活的丝氨酸/苏氨酸激酶。AKT蛋白具有三种亚型AKT 1（PKB α）、AKT 2（PKB β）和AKT 3（PKB γ），它们具有大于85%的序列同一性并且具有相同的结构组织。AKT 2原癌基因在卵巢癌中扩增。在卵巢癌细胞和肿瘤中也经常检测到AKT 1磷酸化和AKT 1激酶活性升高。AKT改变的卵巢癌患者似乎预后不良。由于AKT可以提供保护细胞免受抗癌药物或应激诱导的凋亡的存活信号，因此开发针对AKT的有效和选择性抑制剂是治疗卵巢癌的有吸引力的治疗策略。通过基于结构的设计和筛选，我们已经确定了一个有效的和选择性的AKT小分子抑制剂（称为API-59）。在这个提议中，我们建议评估API-59在表达升高的AKT活性的卵巢癌细胞中的效力、选择性和分子机制。为落实这一概念，将研究以下具体目标： 1.评价API-59对卵巢癌细胞中AKT通路的作用。 1.1.合成API-59。 1.2.评价API-59在表达组成型活性AKT的卵巢癌细胞系中诱导细胞凋亡。 1.3.确定API-59对AKT激酶活性和AKT下游靶标的影响。 1.4.评价API-59对缺乏组成型活性AKT的正常细胞的作用。 2.检查API-59与细胞毒性剂组合在卵巢癌细胞中的作用。 2.1.测试API-59与顺铂组合对诱导细胞凋亡和抑制表达组成型活性AKT的卵巢癌细胞系中的AKT途径的作用。 2.2.检查API-59与紫杉醇组合对诱导细胞凋亡和抑制表达组成型活性AKT的卵巢癌细胞中的AKT途径的抑制作用。 3.体内裸鼠肿瘤模型中API-59的功效评价。 确定API-59处理抑制已建立的小鼠肿瘤中升高的AKT活性和延缓裸鼠中已建立的肿瘤生长的功效。