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）：该研究项目将集中于卵巢癌细胞中新型Akt选择性小分子抑制剂的评估以及该抑制剂对AKT下游靶标的影响。 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蛋白具有三种同工型AKT1（PKBALPHA），AKT2（PKBBETA）和AKT3（PKBGAMMA），它们具有大于85％的序列身份并且具有相同的结构组织。 Akt2原始癌基因在卵巢癌中被扩增。在卵巢癌细胞和肿瘤中，还经常检测到AKT1磷酸化和Akt1激酶活性升高。 AKT改变的卵巢癌患者的预后似乎很差。由于AKT可能会提供一个生存信号，该信号可保护细胞免受抗癌药物或应力诱导的凋亡，因此靶向AKT的有效抑制剂的发展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，我们确定了AKT的有效和选择性的小分子抑制剂（称为API-59）。在此提案中，我们建议评估表达Akt活性升高的卵巢癌细胞中API-59的效力，选择性和分子机制。将研究以下特定目标以解决此概念： 1。评估API-59对卵巢癌细胞中AKT途径的影响。 1.1。 API-59的合成。 1.2。评估API-59在卵巢癌细胞系中诱导凋亡的诱导，这些细胞表达了组成型活性Akt。 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对缺乏组成性活性AKT的正常细胞的影响。 2。检查API-59与卵巢癌细胞中细胞毒性剂的作用。 2.1。测试API-59与顺铂联合诱导凋亡的影响并抑制卵巢癌细胞系中表达组成性活性AKT的AKT途径。 2.2。检查API-59与紫杉醇对诱导细胞凋亡的抑制作用，并抑制卵巢癌细胞中表达组成性活性AKT的AKT途径。 3。评估API-59在体内裸小鼠肿瘤模型中的疗效。 确定API-59治疗抑制已建立小鼠肿瘤中AKT活性升高的功效，并阻止裸鼠已建立的肿瘤的生长。