Long Acting VEGF Inhibitor for Treating RA

用于治疗 RA 的长效 VEGF 抑制剂

• 批准号: 6832460
• 负责人: MARY S. ROSENDAHL
• 金额: $ 10万
• 依托单位: BOLDER BIOTECHNOLOGY, INC.
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-15 至 2005-09-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6832460
• 关键词: angiogenesis inhibitors; arthritis therapy; biotechnology; biotherapeutic agent; cell line; cell proliferation; chemical substitution; drug design /synthesis /production; growth factor receptors; intravenous administration; laboratory rat; molecular cloning; peptide analog; pharmacokinetics; polyethylene glycols; protein engineering; protein purification; recombinant proteins; rheumatoid arthritis; site directed mutagenesis; slow release drug; vascular endothelial growth factors; vascular endothelium

DESCRIPTION (provided by applicant): Rheumatoid arthritis (RA) is a chronic inflammatory disease that causes progressive joint destruction, deformities and disability. One of the earliest observed features of RA is the development of a new vascular network within the synovium that allows for the delivery of cells and nutrients to the invading pannus. This formation of new blood vessels (also known as angiogenesis) is a highly regulated process under the influence of the host microenvironment and a number of secreted mediators. Many proangiogeneic mediators are expressed in RA. One of these mediators in particular, vascular endothelial growth factor (VEGF) appears to play a critical role in the differentiation of endothelial cells and the development of the vascular system in the synovial lining of the joints and therefore, is a reasonable target for therapeutic intervention in RA patients. Our overall goals are to create a long acting VEGF inhibitor and test its efficacy in animal models of RA. Using the published structural information for soluble VEGF Receptor I (also known as sFlt-1), we will rationally design polyethylene glycol (PEG)-sFlt-1 conjugates using cysteine-reactive PEGs. We will introduce a new "free" cysteine using site-directed mutagenesis in regions of sFlt-1 that are believed to be non-essential for biological activity. The "free" cysteine residue will serve as the site for the covalent modification of the protein using a thiol-reactive PEG. This technology allows for the creation of novel, fully active PEG-Cys-sFlt-1 analogues of defined structure and overcomes the problems of reduced bioactivity and heterogeneity when proteins are modified using standard amine-reactive PEGs. During Phase I we will identify sites in sFlt-1 that can be modified without affecting the protein's in vitro bioactivity. We will also perform a pharmacokinetic study to verify that PEGylation extends the circulating half-life of sFlt-1. During Phase II, we develop a cost effective manufacturing process and produce sufficient quantities of PEGylated recombinant sFlt-1 for testing in animal disease models and biochemical characterization.

描述（由申请人提供）： 类风湿性关节炎 (RA) 是一种慢性炎症性疾病，会导致进行性关节破坏、畸形和残疾。 RA 最早观察到的特征之一是滑膜内新血管网络的形成，该网络允许将细胞和营养物质输送到侵入的血管翳。新血管的形成（也称为血管生成）是在宿主微环境和许多分泌介质的影响下高度调控的过程。许多促血管生成介质在 RA 中表达。尤其是这些介质之一，血管内皮生长因子（VEGF）似乎在内皮细胞的分化和关节滑膜内层血管系统的发育中发挥着关键作用，因此，它是 RA 患者治疗干预的合理目标。我们的总体目标是创造一种长效 VEGF 抑制剂并在 RA 动物模型中测试其疗效。利用已发表的可溶性 VEGF 受体 I（也称为 sFlt-1）的结构信息，我们将使用半胱氨酸反应性 PEG 合理设计聚乙二醇 (PEG)-sFlt-1 缀合物。我们将在 sFlt-1 被认为对生物活性非必需的区域中使用定点诱变引入一种新的“游离”半胱氨酸。 “游离”半胱氨酸残基将作为使用硫醇反应性 PEG 对蛋白质进行共价修饰的位点。该技术可以创建具有明确结构的新型、完全活性的 PEG-Cys-sFlt-1 类似物，并克服了使用标准胺反应性 PEG 修饰蛋白质时生物活性降低和异质性降低的问题。在第一阶段，我们将确定 sFlt-1 中可以进行修饰而不影响蛋白质体外生物活性的位点。我们还将进行药代动力学研究，以验证聚乙二醇化可延长 sFlt-1 的循环半衰期。在第二阶段，我们开发了一种具有成本效益的制造工艺，并生产足够数量的聚乙二醇化重组 sFlt-1，用于动物疾病模型测试和生化表征。