STEF as a novel antitumor agent

STEF 作为新型抗肿瘤剂

• 批准号: 7140093
• 负责人: CARL A LUER
• 金额: $ 15.41万
• 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/7140093
• 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）： 一种对多种不同肿瘤细胞系具有强效抗肿瘤活性的天然产物存在于短平头鲨（Sphyrna tiburo）上生殖细胞短期培养物的培养基中。初步数据表明，抑制因子是一种蛋白质，暂时称为Sphyrna tiburo epigonal蛋白-1（STEP-1），它似乎通过阻断DNA合成和诱导增殖细胞凋亡发挥作用。这个项目的广泛的长期目标是分离活性因子并将其纯化成可以通过重组技术生产的形式。目前项目的具体目标是表征STEP-1的分子作用机制并纯化具有生物活性的STEP-1。纯化后，将表征STEP-1的效力和分子作用机制，并评价毒性、药代动力学和体内抗肿瘤活性。生物活性STEP-1的纯化将通过采用各种生物化学和免疫亲和纯化策略来完成，这些策略将利用称为流式细胞术高含量筛选（FC-HCS）的新型高通量流式细胞术筛选技术。还将获得纯化STEP-1的蛋白质序列信息。用于确定STEP-1作用机制的方法包括流式细胞术、免疫印迹、北方印迹和细胞凋亡和细胞周期相关蛋白的其他分子分析。纯化STEP-1蛋白的毒性和药代动力学将在CFU测定中使用造血祖细胞进行评价，并按照标准ISO方法在一系列标准临床前毒性测定中进行测试。将在注射STEP-1的小鼠中进行药理学研究。如果这些研究证明STEP-1具有有效的抗肿瘤活性，具有可接受的毒性和药代动力学特性，则将为继续进行未来研究提供理由，这些研究旨在生产重组STEP-1，以在I期临床试验中作为抗癌剂进行进一步测试。