PTPases as Therapeutic Targets

PTPases 作为治疗靶点

• 批准号: 6948210
• 负责人: Taolin Yi
• 金额: $ 27.23万
• 依托单位: CLEVELAND CLINIC LERNER COM-CWRU
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-09-30 至 2007-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6948210
• 关键词: antineoplastics; athymic mouse; carcinogenesis inhibitor; cell line; chemical group; drug discovery /isolation; enzyme activity; enzyme substrate complex; enzyme therapy; gene expression; gene mutation; in situ hybridization; mass spectrometry; metastasis; neoplasm /cancer therapy; nuclear magnetic resonance spectroscopy; pathologic process; pentamidine; phosphatase inhibitor; polymerase chain reaction; protein structure; protein tyrosine phosphatase

DESCRIPTION (provided by applicant): PRL family phosphatases (PRL-1, PRL-2 and PRL-3) are highly attractive targets for developing inhibitors as novel anti-cancer therapeutics because their over-expression has oncogenic effects and plays a potentially pathogenic role in human malignancies. Our recent studies provided the first evidence that pentamidine (PE), an anti-protozoan drug with an unknown mechanism of action, is an effective inhibitor of PRLs and has anti-cancer effects. The goal of this study is to test our hypothesis that PE has growth inhibitory effects against tumor cells via inactivating PRLs through direct binding to the PTPases and has therapeutic potential against malignancies associated with PRL over-expression. Aim 1. To evaluate the roles of PRL PTPases in mediating PE anti-cancer effects, we will 1) determine the capacities of Peinsensitive mutants of PRL-1, PRL-2 and PRL-3 individually and in combinations to confer resistance to Peinduced growth inhibition in WM9 human melanoma cell line to define the contributions of each PRL in PE effects against the cancer cells and in cancer cell growth; 2) determine PE activity to reverse transformed phenotypes induced by over-expression of PRLs or PTPalpha oncogene in NIH3T3 cells to further elucidate PE anti-cancer mechanism; 3) determine PE effects on lung metastasis of A549 transfectants of PRLs in nude mice to provide prove of concept evidence regarding PE anti-metastasis activity and the roles of individual PRLs in tumor metastasis; 4) identify human malignancies over-expressing PRLs by RNA in situ hybridization to assess the significance of individual PRLs as anti-cancer targets. Aim 2. To elucidate inhibition mechanism of PE against PRLs, we will 1) identify key chemical groups in PE essential for PRL inhibition and PTPase specificity by evaluating 10 PE-derivatives in which PE chemical groups have been selectively modified; 2) characterize PE / PRL complex formation and PE effects on PRL-1/substrate interaction by mass spectrometry and biochemical analysis; 3) analyze solution structures of PRLs and their complexes with PE by NMR. Results of the study will provide proof of principle evidence to establish PE as a PRL-targeted agent with anti-cancer potential, reveal the inhibition mechanism of PE against PRLs at molecular levels, provide insights for rational design and development of more specific and less toxic novel PRL inhibitors and identify human malignancies potentially responsive to PRL-targeted therapy.

描述（由申请人提供）：PRL家族磷酸酶（PRL-1，PRL-2和PRL-3）是将抑制剂作为新型抗癌治疗剂开发抑制剂的高度吸引力的靶标，因为它们的过表达具有致癌作用，并且在人类恶性肿瘤中起潜在的致病作用。我们最近的研究提供了第一个证据，即一种有效的PRL抑制剂，具有抗癌作用，是一种具有未知作用机理的抗植物药物。这项研究的目的是检验我们的假设，即通过直接结合PTPases，PE对肿瘤细胞具有生长抑制作用，并具有与PRL过表达相关的恶性肿瘤的治疗潜力。目的1。为了评估PRL PTPases在介导PE抗癌作用中的作用，我们将确定单独的PRL-1，PRL-2和PRL-3的Peinsentive突变体的能力，并组合以赋予对WM9人类瘤细胞中PE癌的癌细胞的生长抑制作用，以赋予对癌细胞的生长抑制作用，以定义PEL的癌症癌症效应，并在癌症中造成癌症效应，并在癌症中效应。 2）确定由NIH3T3细胞中PRLS或PTPALPHA癌基因过表达引起的PE活性，以逆转转化的表型，以进一步阐明PE抗癌机制； 3）确定对裸鼠A549转染剂的肺转移剂对肺转移的影响，以提供有关PE抗中端活性的概念证据，以及单个PRL在肿瘤转移中的作用； 4）通过RNA通过原位杂交确定过度表达PRL的人类恶性肿瘤，以评估单个PRL作为抗癌靶标的重要性。目的2。为了阐明PE对PRL的抑制机制，我们将通过评估10种选择性修饰PE化学基团的10个PE衍生品，确定PE中PE中的关键化学基团对于PRL抑制和PTPase特异性必不可少的； 2）通过质谱和生化分析表征PE / PRL复合物的形成和PE对PRL-1 /底物相互作用的影响； 3）分析NMR与PE的PRL及其复合物的溶液结构。该研究的结果将提供主要证据证明PE作为具有抗癌潜力的PRL靶向药物的证据，揭示了PE在分子水平上对PRL的抑制作用机制，为理性设计和开发更具体且毒性较小的新型PRL抑制剂提供了见解，并确定人类恶性肿瘤对PRL含量的治疗有潜在地响应。