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: 6703347
• 负责人: Jiayuh Lin
• 金额: $ 13.39万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 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 2004-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6703347
• 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）：该研究项目将重点评估一种新型 AKT 选择性小分子抑制剂在卵巢癌细胞中的作用以及该抑制剂对 AKT 下游靶点的影响。 AKT 或蛋白激酶 B (PKB) 是一种丝氨酸/苏氨酸激酶，可响应生长因子或细胞因子而激活。 AKT 蛋白具有 AKT1 (PKBalpha)、AKT2 (PKBbeta) 和 AKT3 (PKBgamma) 三种同工型，它们具有大于 85% 的序列同一性并具有相同的结构组织。 AKT2 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 AKT1 磷酸化和 AKT1 激酶活性升高。 AKT 改变的卵巢癌患者似乎预后较差。由于 AKT 可以提供保护细胞免受抗癌药物或应激诱导的细胞凋亡的生存信号，因此开发针对 AKT 的有效选择性抑制剂是治疗卵巢癌的一种有吸引力的治疗策略。通过基于结构的设计和筛选，我们鉴定出了一种有效且选择性的 AKT 小分子抑制剂（称为 API-59）。在本提案中，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。为了解决这一概念，将研究以下具体目标： 1.评价API-59对卵巢癌细胞AKT通路的影响。 1.1. API-59的合成。 1.2.评估 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的情况。 1.3.确定 API-59 对 AKT 激酶活性和 AKT 下游靶标的影响。 1.4.评估 API-59 对缺乏组成型活性 AKT 的正常细胞的影响。 2.检查API-59与细胞毒剂联合对卵巢癌细胞的作用。 2.1.测试 API-59 联合顺铂在表达组成型活性 AKT 的卵巢癌细胞系中诱导细胞凋亡和抑制 AKT 通路的效果。 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 AKT 通路的抑制作用。 3. API-59在裸鼠肿瘤模型体内的疗效评价。 确定 API-59 治疗抑制已形成小鼠肿瘤中 AKT 活性升高并延缓裸鼠已形成肿瘤生长的功效。