Novel Glioma uPA Inhibitor Design Guided by Intracellular signaling Pathways

细胞内信号通路引导的新型胶质瘤 uPA 抑制剂设计

基本信息

批准号：
7656163
负责人：
FREDRIC A GORIN
金额：
$ 36.13万
依托单位：
UNIVERSITY OF CALIFORNIA AT DAVIS
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-09-01 至 2011-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7656163
关键词：
Adult Affect Alkylating Agents Amiloride Animals Anoikis Apoptosis Apoptotic Autocrine Communication Autopsy Behavioral Blinded Body Weight Brain Brain Neoplasms Breast Cancer Cell Cancer cell line Cell Adhesion Cell Death Cell Line Cell Proliferation Cell Survival Cell membrane Cell surface Cells Cessation of life Coupled Cytostatics Data Dose Drug Delivery Systems Drug vehicle Effectiveness Equilibrium Evaluation FDA approved Glioma Growth Homeostasis Human Hypoxia Image Immunodeficient Mouse Implant In Vitro Individual Invaded Kinetics Lead Luciferases MAPK14 gene Malignant Glioma Malignant Neoplasms Malignant neoplasm of brain Measurement Measures Mediating Memory Messenger RNA Methods Modeling Molecular Monitor Motor Mus Normal tissue morphology Nude Mice Oligonucleotides Organ Outcome Pathology Pathway interactions Peptide Hydrolases Pharmaceutical Preparations Plasminogen Inactivators Poison Population Proliferating Property Proteins Protocols documentation RNA RNA Interference Radiation therapy Rattus Recurrence Research Resistance Seizures Series Signal Pathway Signal Transduction Small Interfering RNA Stress Subfamily lentivirinae Surface Surveys System Time Titrations Toxic effect Transfection Treatment Efficacy Treatment Protocols Urokinase Vascular Endothelial Growth Factors Xenograft Model Xenograft procedure abstracting base brain cell cancer cell cell type cytotoxic cytotoxicity design dosage drug candidate drug mechanism extracellular glioma cell line immunocytochemistry in vivo indexing inhibitor/antagonist killings migration mouse model neoplastic cell neovascularization neurobehavioral neuropathology neurotoxicity novel novel therapeutics prevent research study response small molecule synergism temozolomide tumor tumor growth

项目摘要

1. Abstract. We synthesized a novel set of non-toxic compounds that kill proliferating and nonproliferating glioma cells residing in perinecrotic tumor microenvironments. These hypoxic glioma populations have increased resistance to conventional radiation therapy coupled with alkylating agents such that cancer recurs in greater than 90% of individuals with high grade gliomas. The design and syntheses of these novel compounds were derived from our recent demonstration that urokinase plasminogen activator (uPA) becomes enzymatically activated intracellularly in hypoxic and acidotic high grade human gliomas. Amiloride is an FDA-approved drug that selectively inhibits uPA, but not other proteases. We synthesized and evaluated a series of amiloride-based compounds that inhibit either total or extracellular uPA activity in human glioma cells. Interestingly, forms of the drugs excluded from the cell interior and act only on surface uPA are cytostatic. Significantly, congeners that permeate the plasma membrane and also inhibit intracellular uPA trigger glioma cell death, yet do not affect normal brain cell types. Our pharmacological findings are consistent with RNA interference experiments demonstrating that inhibition of uPA mRNA initiates apoptosis of glioma cells by unknown cellular mechanisms. These observations point to the novel notion that infiltrative, hypoxic tumor cells can become reliant on intracellular uPA for their survival. Since intracellular uPA activation is not observed in normal tissue cell types or in normal adult brain, these observations suggest that intracellular uPA may represent an important drug target of malignant glioma cells that survive and recur in hypoxic-ischemic microenvironments. The selective anti-glioma cytotoxicities and absence of CNS toxicities of our lead compounds in rat orthotopic glioma xenografts are encouraging given the extremely poor outcome of most individuals having high grade gliomas. Aim 1 will investigate the in vivo efficacies of the lead compounds in a NOD/SCID-gamma murine intracerebral glioma xenograft model. We will investigate the efficacies of our lead compounds on {1} primary glial tumor growth kinetics and on {2} recurrent glioma following radiation therapy and temozolamide (TMZ) treatment. Because uPA inhibition impedes glioma neovascularization, we will also investigate {3} the potential synergism of our lead compounds to prevent or retard the previously described disseminated intracerebral growth of VEGF-depleted glioma cells in mice. Aim 2 will utilize these novel small molecule uPA inhibitors to identify the intracellular mechanisms contributing to their selective anti-glioma cytotoxicity. We will also target total uPA using RNA interference and compare resultant glioma cell death pathways to those identified using our small molecule inhibitors.

1。摘要。我们合成了一组新型的无毒化合物，这些化合物杀死了居住在围产肿瘤微环境中的增殖和非增殖神经胶质瘤细胞。这些低氧神经胶质瘤种群对常规放射疗法的抵抗力增加，并与烷基化剂相结合，使得大于90％的高级神经胶质瘤个体的癌症复发。这些新型化合物的设计和合成源自我们最近的证明，即尿激酶纤溶酶原激活剂（UPA）在缺氧和酸性高级人神经胶质瘤中酶细胞内酶活性。 Amiloride是一种由FDA批准的药物，有选择地抑制UPA，但没有其他蛋白酶。我们合成并评估了一系列基于阿米洛利的化合物，这些化合物抑制人神经胶质瘤细胞中的总体或细胞外UPA活性。有趣的是，从细胞内部排除的药物的形式仅在表面UPA上起作用。值得注意的是，渗透到质膜并抑制细胞内UPA触发神经胶质瘤细胞死亡的同源物，但不会影响正常的脑细胞类型。我们的药理发现与RNA干扰实验一致，表明抑制UPA mRNA会通过未知的细胞机制启动神经胶质瘤细胞的凋亡。这些观察结果表明，浸润性，低氧肿瘤细胞可以依靠细胞内UPA的生存依赖的新观念。由于在正常组织细胞类型或正常的成年大脑中未观察到细胞内UPA激活，因此这些观察结果表明，细胞内UPA可能代表了恶性神经胶质瘤细胞的重要药物靶标，该靶标在低氧 - 缺血性微环境中存活并复发。鉴于大多数患有高级别的神经胶质瘤的人的结果极差，我们在大鼠原位神经胶质瘤异种移植物中的选择性抗脱胶瘤细胞毒性和我们铅化合物的中枢神经系统毒性的缺失令人鼓舞。 AIM 1将研究点头/scid-gamma鼠内膜神经胶质瘤模型中铅化合物的体内效率。我们将研究铅化合物对{1}原发性神经胶质肿瘤生长动力学的有效性以及放射治疗和替莫唑胺（TMZ）治疗后{2}复发性神经胶质瘤的效率。因为UPA抑制阻碍了神经胶质瘤新血管形成，我们还将研究{3}我们的铅化合物的潜在协同作用，以预防或延迟小鼠VEGF缺失的神经胶质瘤细胞的前描述的脑内生长。 AIM 2将利用这些新型的小分子UPA抑制剂来识别有助于其选择性抗胶质瘤细胞毒性的细胞内机制。我们还将使用RNA干扰靶向总UPA，并将结果胶质瘤细胞死亡途径与使用我们的小分子抑制剂鉴定的胶质瘤细胞死亡途径。