Evaluation of a novel AKT inhibitor in ovarian cancer

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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.