Manufacturing of Trojan Horse-TNFR Decoy Receptor Fusion Protein

特洛伊木马-TNFR诱饵受体融合蛋白的制造

基本信息

批准号：
8307104
负责人：
RUBEN J. BOADO
金额：
$ 14.85万
依托单位：
ARMAGEN TECHNOLOGIES, INC.
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-04-01 至 2012-09-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8307104
关键词：
A Mouse Affinity Affinity Chromatography Alzheimer&apos s Disease Anions Binding Binding Sites Biochemical Bioreactors Blood - brain barrier anatomy Brain Brain Diseases Cations Cell Line Cells Cerebrum Chimeric Proteins Chinese Hamster Chinese Hamster Ovary Cell Chromatography Clinical Trials Cloning Development Dose Drug Kinetics Engineering Epitopes Equus caballus Etanercept Extracellular Domain Future Goals Human Human Engineering Immunoglobulin G Inflammatory Insulin Insulin Receptor Macaca mulatta Mediating Mental Depression Methodology Monoclonal Antibodies Ovary Parkinson Disease Peripheral Pharmaceutical Preparations Pharmacology Phase Plasma Play Primates Production Proteins Receptors, Tumor Necrosis Factor, Type II Research Role Safety Scheme Sepharose Serum-Free Culture Media Small Business Innovation Research Grant Staging Structure Surface Plasmon Resonance TNF gene Testing Therapeutic Tissues Toxicology Tumor Necrosis Factor Receptor Tumor Necrosis Factor-alpha Validation Work cytokine design drug development human INSR protein human TNF protein in vivo inhibitor/antagonist intravenous administration meetings molecular trojan horse mouse model novel receptor receptor binding scale up small molecule uptake

项目摘要

DESCRIPTION (provided by applicant): Tumor necrosis factor (TNF)-¿ plays a pro-inflammatory role in brain diseases. The biologic TNF inhibitors (TNFI), such as the TNF decoy receptor cannot be developed for brain diseases, because the TNFIs are large molecules that do not cross the blood-brain barrier (BBB). The present work continues the drug development of a re-engineered form of the human type II TNF receptor (TNFR), wherein the TNFR 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-TNFR fusion protein acts as a molecular Trojan horse to ferry the fused decoy receptor 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-TNFR fusion protein. The proposed phase I SBIR research will develop a manufacturing scheme for production of the HIRMAb-TNFR 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 small scale manufacturing methodology developed in phase I will then be scaled up in phase II to the 50L bioreactor stage for manufacturing of the HIRMAb-TNFR fusion protein at production levels that can support future clinical trials. PUBLIC HEALTH RELEVANCE: Biologic tumor necrosis factor inhibitors (TNFI), such as the tumor necrosis factor (TNF) decoy receptor, cannot be developed for brain diseases, because these large molecule drugs do not cross the blood-brain barrier (BBB). The present research will re-engineer the human type II TNF receptor (TNFR) as an IgG-TNFR fusion protein, where the IgG part is a genetically engineered monoclonal antibody that crosses the BBB via transport on the endogenous insulin receptor. The IgG acts as a molecular Trojan horse to ferry across the BBB the TNFI.

描述（由申请人提供）：肿瘤坏死因子（TNF）-¿ TNF 诱饵受体等生物 TNF 抑制剂 (TNFI) 在脑部疾病中发挥促炎作用，因为 TNFI 是不能穿过血脑屏障 (BBB) 的大分子。目前的工作继续进行重新设计的人类 II 型 TNF 受体 (TNFR) 的药物开发，因此 TNFR 是作为 IgG 融合蛋白生产的。IgG 部分是一种基因工程单克隆抗体。 HIRMAb-TNFR 融合蛋白的 HIRMAb 部分充当分子特洛伊木马，通过内源 BBB 胰岛素受体上的受体介导的转运将融合的诱饵受体运送穿过 BBB。本研究的预 SBIR 可行性阶段描述了 HIRMAb-TNFR 融合蛋白的工程、表达、生化验证以及恒河猴体内血浆药代动力学和 BBB 转运。拟议的 I 期 SBIR 研究将开发 HIRMAb-TNFR 融合蛋白的生产方案。该生产方案旨在生产符合 FDA 在纯度、效力、安全性和杂质方面的规格的治疗产品，以便生产。可以在未来融合蛋白的 GMP 生产中复制用于临床试验，第一阶段开发的小规模生产方法将在第二阶段扩大到 50L 生物反应器阶段，用于生产 HIRMAb-TNFR。生产水平的融合蛋白可以支持未来的临床试验。公共健康相关性：生物肿瘤坏死因子抑制剂 (TNFI)，例如肿瘤坏死因子 (TNF) 诱饵受体，不能用于治疗脑部疾病，因为这些大分子药物不能穿过现有的血脑屏障 (BBB)。研究将人类 II 型 TNF 受体 (TNFR) 重新设计为 IgG-TNFR 融合蛋白，其中 IgG 部分是一种基因工程单克隆抗体， IgG 通过内源性胰岛素受体转运穿过 BBB，充当分子特洛伊木马，运送 TNFI 穿过 BBB。