Targeting the VEGF receptor with specific proteolysis

通过特异性蛋白水解作用靶向 VEGF 受体

• 批准号: 6740950
• 负责人: SANDRA W RUGGLES
• 金额: $ 10万
• 依托单位: CATALYST BIOSCIENCES
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-04-01 至 2005-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6740950
• 关键词: apoptosis; cell surface receptors; clinical research; combinatorial chemistry; enzyme linked immunosorbent assay; enzyme substrate; growth factor receptors; intracellular; peptide library; protein engineering; protein structure function; proteolysis; receptor binding; receptor expression; serine proteinases; technology /technique development; vascular endothelial growth factors

DESCRIPTION (provided by applicant): The goal of this project is to combine a powerful protease profiling technology with protein engineering to develop a designer protease with novel extended substrate specificity for the treatment of cancer. Inactivation of signaling through the VEGFR-2/FIk-1 receptor has a potent anti-angiogenic effect on tumor vasculature, leading to the systematic death of the tumor. Current small molecule and antibody efforts have been unable to fully inhibit the VEGF signaling in clinical trials. Thus the inhibition of angiogenic signaling through FIk-1 represents an underdeveloped therapeutic area. Due to their catalytic nature and smaller size, engineered proteases as a therapeutic modality have a number of advantages over competing platforms including better tumor penetration, better target saturation, higher effectiveness, and potentially lower dosing. By starting with natural proteases that have high serum half-lives and reduced inhibitor binding, we will design a serine protease with an in vitro specificity that matches the FIk-1 stalk over a six amino acid region using the powerful pairing of protein engineering techniques and a proprietary protease substrate profiling technology. Positional scanning synthetic combinatorial library (PSSCL) profiling allows one to generate a complete substrate specificity profile or "fingerprint" of a protease using a single 96-well plate. PSSCL can be used to track the change in the specificity profile of variant proteases. Therefore, therapeutically relevant second generation molecules can be identified which have enhanced specificity toward target substrates and diminished specificity towards alternate substrates. Variant proteases will be extensively tested for their ability to effectively hydrolyze FIk-1, abrogate intracellular signaling, and reduce VEGF levels through binding of the soluble receptor fragment. The best candidate for clinical development will be an engineered protease that irreversibly inactivates the target receptor leading to a potent anti-angiogenic effect.

描述（由申请人提供）：该项目的目的是将强大的蛋白酶分析技术与蛋白质工程结合起来，以开发具有新型扩展底物特异性以治疗癌症的设计师蛋白酶。通过VEGFR-2/FIK-1受体失活的信号传导对肿瘤脉管系统具有有效的抗血管生成作用，导致肿瘤的系统死亡。当前的小分子和抗体工作一直无法完全抑制临床试验中的VEGF信号传导。因此，通过FIK-1抑制血管生成信号是一个欠发达的治疗区域。由于其催化性质和较小的尺寸，工程蛋白酶作为治疗方式比竞争平台具有许多优势，包括更好的肿瘤渗透，更好的靶饱和度，更高的效率和潜在的剂量。 通过从具有高血清半衰期和降低抑制剂结合的天然蛋白酶开始，我们将使用体外特异性设计一种丝氨酸蛋白酶，该蛋白酶使用蛋白质工程技术的强大配对和专有的蛋白酶蛋白酶底物分析技术与六氨基酸区域的FIK-1茎相匹配。位置扫描合成组合库（PSSCL）谱分析使人们可以使用单个96孔板生成蛋白酶的完整底物特异性曲线或“指纹”。 PSSCL可用于跟踪变体蛋白酶的特异性曲线的变化。因此，可以鉴定出与治疗相关的第二代分子，从而增强了对靶标底物的特异性，并降低了对交替底物的特异性。变体蛋白酶将通过可溶性受体片段的结合来有效地水解Fik-1，废除细胞内信号传导并降低VEGF水平的能力进行广泛测试。临床发育的最佳候选者将是一种工程蛋白酶，可不可逆转地使靶受体失活，从而产生有效的抗血管生成作用。