Angiociden, A New Angiogenesis Inhibitor

Angiociden，一种新的血管生成抑制剂

• 批准号: 6614101
• 负责人: GEORGE Paul TUSZYNSKI
• 金额: $ 10.72万
• 依托单位: TEMPLE UNIV OF THE COMMONWEALTH
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-07-01 至 2005-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6614101
• 关键词: SCID mouse; affinity chromatography; angiogenesis inhibitors; breast neoplasms; cell adhesion; cell proliferation; enzyme linked immunosorbent assay; extracellular matrix; growth factor; human tissue; immunocytochemistry; lung neoplasms; neoplasm /cancer chemotherapy; neoplasm /cancer invasiveness; nonhuman therapy evaluation; protein protein interaction; recombinant proteins; thrombospondins; vascular endothelium

DESCRIPTION (provided by applicant): We have cloned a novel high affinity novel thrombospondin-1 (TSP-1) binding protein from a prostate cancer cell library. The recombinant protein expressed in bacteria, termed angiocidin, inhibited endothelial cell viability and tube formation in vitro and prevented the growth of tumors in vivo. When injected intravenously into mice bearing Lewis Lung carcinoma, the protein inhibited the growth of the tumor by more than 500 percent as compared to the buffer control. As shown by immunohistochemical staining of human breast tumors, angiocidin was expressed in malignant ductal epithelium and tumor microvasculature while no significant expression was observed in normal and benign tissues. Our preliminary data indicate that angiocidin is posttranslationally modified in eukaryotic cells. Post-translational modification such as glycosylation are known to greatly affect the half-life of proteins in circulation and may therefore increase the bioavailability and half-life of angiocidin injected systemically. Thus, these post-translational changes may increase the anti-tumor activity of angiocidin as well as produce protein with no endotoxin contamination suitable for clinical development. In this supplemental proposal, we will evaluate which protein expression system is optimal for production of angiocidin suitable for administration in human clinical cancer trials. To accomplish this goal, the anti-tumor activity of recombinant angiocidin expressed in bacteria with angiocidin expressed in yeast, insect cells, and Chinese hamster ovary cells will be compared. Anti-tumor activity of the recombinant proteins expressed in the three expression systems will be evaluated in two animal models, an orthotopic and a syngeneic mouse model, as well as in an in vitro angiogenesis model. Criteria for choosing a protein expression will include specific anti-tumor activity, and protein yield. These experiments will develop the necessary protocols needed for commercial production of protein under GMP (good manufacturing practices) conditions suitable for use in clinical trials

描述（由申请人提供）：我们从前列腺癌细胞文库中克隆了一种新的高亲和力的新型血小板反应蛋白-1（TSP-1）结合蛋白。在细菌中表达的重组蛋白，称为angiocidin，在体外抑制内皮细胞活力和管形成，并在体内阻止肿瘤的生长。当静脉注射到荷刘易斯肺癌的小鼠中时，与缓冲液对照相比，该蛋白质抑制肿瘤生长超过500%。免疫组化结果显示，angiocidin在乳腺癌组织中表达于乳腺癌导管上皮和肿瘤微血管，而在正常乳腺组织和良性乳腺组织中无明显表达。我们的初步数据表明，angiocidin在真核细胞中是后修饰的。已知翻译后修饰（如糖基化）会极大地影响循环中蛋白质的半衰期，因此可能会增加全身注射血管杀菌素的生物利用度和半衰期。因此，这些翻译后的变化可能会增加血管杀菌素的抗肿瘤活性，以及产生适合临床开发的无内毒素污染的蛋白质。在本补充提案中，我们将评估哪种蛋白质表达系统最适合生产适用于人类临床癌症试验的血管杀菌素。为了实现这一目标，将比较在细菌中表达的重组血管杀菌素与在酵母、昆虫细胞和中国仓鼠卵巢细胞中表达的血管杀菌素的抗肿瘤活性。将在两种动物模型（原位和同基因小鼠模型）以及体外血管生成模型中评价在三种表达系统中表达的重组蛋白的抗肿瘤活性。选择蛋白质表达的标准将包括特异性抗肿瘤活性和蛋白质产率。这些实验将开发在GMP（良好生产规范）条件下商业化生产蛋白质所需的必要方案，适用于临床试验