Re-Engineering Blood-Borne Erythropoietin for Targeted Delivery

说明(申请人提供)：促红细胞生成素(EPO)是一种潜在的治疗脑血管疾病(包括中风)的新药。然而，EPO是一种大分子药物，不能穿过脑内毛细血管内皮壁，形成血脑屏障。目前的工作继续药物开发的重新设计形式的EPO，其中EPO是生产的一种免疫球蛋白融合蛋白。Ig G部分是一种针对人胰岛素受体(HIR)的基因工程单抗(MAb)。HIRMAb-EPO融合蛋白的HIRMAb部分作为一个分子特洛伊木马，通过内源性BBB胰岛素受体上的受体介导的转运，将融合的EPO运送到BBB上。本研究的Pre-SBIR可行性阶段描述了HIRMAb-EPO融合蛋白的工程设计、表达、生化验证、体内血浆药代动力学和BBB在恒河猴体内的转运。拟议的第一阶段SBIR研究将开发一种生产HIRMAb-EPO融合蛋白的制造方案。这种制造将被设计成生产一种在纯度、效力、安全性和杂质方面符合FDA规范的治疗产品，以便这种制造可以在未来用于临床试验的融合蛋白的GMP生产中复制。在第一阶段产生的融合蛋白将在第二阶段用于HIRMAb-EPO融合蛋白在恒河猴体内的活性和毒理学。第二阶段的目标是生产一个功效/毒理学数据包，可以提交给FDA，用于设计这种新的免疫球蛋白-促红细胞生成素融合蛋白的未来GLP毒理学。 公共卫生相关性：促红细胞生成素(EPO)是一种组织保护剂，可开发为治疗脑血管疾病(包括中风)的新药。然而，外周给药后EPO不能穿透大脑，因为EPO不能穿过脑内皮细胞壁，形成血脑屏障(BBB)。本研究将开发一种新的免疫球蛋白-促红细胞生成素融合蛋白，用于治疗包括中风在内的脑血管疾病。

DESCRIPTION (provided by applicant): Erythropoietin (EPO) is a potential new pharmaceutical to treat vascular disease of the brain, including stroke. However, EPO is a large molecule pharmaceutical that does not cross the capillary endothelial wall in brain, which forms the blood-brain barrier (BBB). The present work continues the drug development of a re-engineered form of EPO, wherein the EPO is produced as an IgG fusion protein. The IgG part is a genetically engineered monoclonal antibody (MAb) against the human insulin receptor (HIR). The HIRMAb part of the HIRMAb-EPO fusion protein acts as a molecular Trojan horse to ferry the fused EPO across the BBB via receptor-mediated transport on the endogenous BBB insulin receptor. The pre-SBIR feasibility stage of this research describes the engineering, expression, biochemical validation, and in vivo plasma pharmacokinetics and BBB transport in the Rhesus monkey of the HIRMAb-EPO fusion protein. The proposed phase I SBIR research will develop a manufacturing scheme for production of the HIRMAb-EPO fusion protein. This manufacturing will be designed to produce a therapeutic product that meets FDA specifications with regard to purity, potency, safety, and impurities, so that the manufacturing can be replicated in future GMP production of the fusion protein for clinical trials. The fusion protein produced in phase I will then be used in phase II for in vivo activity and toxicology of the HIRMAb- EPO fusion protein in Rhesus monkeys. The goal of phase II is to produce an efficacy/toxicology data package that can be presented to the FDA for design of future GLP toxicology of this new IgG-EPO fusion protein. PUBLIC HEALTH RELEVANCE: Erythropoietin (EPO) is a tissue-protective agent that could be developed as a new drug for the treatment of vascular disorders of the brain, including stroke. However, EPO cannot penetrate the brain following peripheral administration, because EPO does not cross the endothelial wall in brain, which forms the blood-brain barrier (BBB). This research will develop a new IgG-EPO fusion protein for the treatment of vascular disease of the brain including stroke.