STEF as a novel antitumor agent

STEF 作为新型抗肿瘤剂

• 批准号: 6958615
• 负责人: CARL A LUER
• 金额: $ 17.78万
• 依托单位: MOTE MARINE LABORATORY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-06-13 至 2007-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6958615
• 关键词: SDS polyacrylamide gel electrophoresis; animal extract; antineoplastics; apoptosis; biological products; cell cycle; cell line; cell transformation; chromatography; cytotoxicity; drug discovery /isolation; drug resistance; flow cytometry; high throughput technology; neoplastic cell; neoplastic growth; phenotype; protein sequence; proteomics; sharks

DESCRIPTION (provided by applicant): A natural product with potent anti-tumor activity against a variety of different tumor cell lines is present in media from short-term cultures of bonnethead shark (Sphyrna tiburo) epigonal cells. Preliminary data suggest that the inhibitory factor is a protein, tentatively termed Sphyrna tiburo epigonal protein-1 (STEP-1), and that it appears to act by blocking DNA synthesis and inducing apoptosis in proliferating cells. The broad, long-term Objectives of this project are to isolate the active factor and purify it to a form that can be produced through recombinant technology. The specific aims of the current project are to characterize the molecular mechanism of action of STEP-1 and to purify bioactive STEP-1. Once purified, the potency and molecular mechanism of action of STEP-1 will be characterized, and the toxicity, pharmacokinetics, and the in-vivo anti-tumor activity will be evaluated. Purification of bioactive STEP-1 will be accomplished by employing various biochemical and immunoaffinity purification strategies that will utilize a novel high throughput flow cytometric screening technique termed Flow Cytometric High Content Screening (FC-HCS). Protein sequence information of purified STEP-1 will also be obtained. Methods used to determine the mechanism of action of STEP-1 will include FACS, immunoblot, Northern blot and other molecular analyses of apoptosis and cell cycle associated proteins. Toxicity and pharmacokinetics of purified STEP-1 protein will be evaluated using hematopoietic progenitors in CFU assays and testing in a battery of standard preclinical toxicity assays following standard ISO methods. Pharmacologic studies will be performed in mice injected with STEP-1. If these studies demonstrate that STEP-1 has potent anti-tumor activity with acceptable toxicity and pharmacokinetic properties, it will provide the justification to proceed with future studies involving aimed at producing recombinant STEP-1 for further testing as an anti-cancer agent in phase I clinical trials.

描述（由申请人提供）： 培养基中，介质中存在着具有强大抗肿瘤活性的天然抗肿瘤活性的天然产物，来自邦纳特鲨鱼（Sphyrna tiburo）表位细胞的短期培养物。初步数据表明，抑制因子是一种蛋白质，暂定称为sphyrna tiburo表位蛋白-1（步骤1），并且似乎通过阻断DNA合成并在增殖细胞中诱导凋亡来起作用。该项目的广泛，长期目标是隔离活性因子，并将其纯化为可以通过重组技术产生的形式。当前项目的具体目的是表征步骤1的分子作用机理并净化生物活性步骤1。一旦纯化，将表征步骤1的作用的效力和分子机理，并将评估毒性，药代动力学和体内抗肿瘤活性。生物活性步骤1的纯化将通过采用各种生物化学和免疫亲和力纯化策略来实现，这些策略将利用一种新型的高通量流式细胞量筛选技术称为流式细胞仪高含量筛选（FC-HCS）。还将获得纯化步骤1的蛋白质序列信息。用于确定步骤1的作用机理的方法将包括FACS，免疫印迹，北印迹和其他分子分析和细胞周期相关蛋白的分子分析。将在CFU测定中使用造血祖细胞评估纯化的Step-1蛋白的毒性和药代动力学，并根据标准ISO方法进行一系列标准的临床前毒性测定法进行测试。在注射步骤1的小鼠中将进行药理学研究。如果这些研究表明，步骤1具有具有可接受的毒性和药代动力学特性的有效抗肿瘤活性，则将提供合理的理由，以进行未来的研究，涉及旨在在I期临床试验中作为抗癌剂进一步测试重组STEP-1。