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

• 批准号: 8627527
• 负责人: RUBEN J. BOADO
• 金额: $ 58.95万
• 依托单位: ARMAGEN TECHNOLOGIES, INC.
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-03-01 至 2016-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8627527
• 关键词: Adult; Affinity Chromatography; Alzheimer' s Disease; Anion Exchange Resins; Anions; Antibodies; Arthritis; Binding; Biochemical; Bioreactors; Blood - brain barrier anatomy; Brain; Brain Diseases; Cation Exchange Resins; Cations; Cells; Cerebrospinal Fluid; Chimeric Proteins; Chinese Hamster; Chinese Hamster Ovary Cell; Chromatography; Chronic; Clinical Trials; Development; Disease; Dose; Drug Impurity; Drug Kinetics; Encephalitis; Engineering; Enzyme-Linked Immunosorbent Assay; Equus caballus; Etanercept; Extracellular Domain; Future; Genetic Engineering; Glucose; Goals; Head; Human; IgG1; Immune response; Immunoglobulin G; Inflammation; Inflammatory; Insulin Receptor; Laboratories; Macaca mulatta; Measures; Mediating; Methodology; Methods; Mono-S; Monoclonal Antibodies; Names; Neurodegenerative Disorders; Organ; Ovary; Parkinson Disease; Peripheral; Pharmaceutical Preparations; Pharmacology; Pharmacology and Toxicology; Phase; Plasma; Play; Primates; Production; Proteins; Receptors, Tumor Necrosis Factor, Type II; Reporting; Research; Role; Safety; Series; Signal Transduction Pathway; Small Business Innovation Research Grant; Staging; Structure; TNF gene; Tail; Temperature; Testing; Time; Toxicology; Tumor Necrosis Factor Receptor; Tumor Necrosis Factor-alpha; Urine; Work; age related; base; cytokine; drug development; glucose tolerance; human INSR protein; human TNF protein; inhibitor/antagonist; manufacturing scale-up; meetings; molecular trojan horse; nano; neuropathology; programs; public health relevance; receptor; relating to nervous system; small molecule

DESCRIPTION (provided by applicant): Tumor necrosis factor (TNF)-alpha plays a pro-inflammatory role in aging-related neurodegenerative diseases, such as Alzheimer's disease or Parkinson's disease. 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 BBB-penetrating biologic TNFI, which is 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-TNFR fusion protein is named AGT-110. 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. In this phase II SBIR project, the methodology for manufacturing of the HIRMAb-TNFR fusion protein at a large scale suitable for production of clinical trial lots of study drug will be developed. The stably transfected host cell will be cultured in a 50L bioreactor, and the fusion protein will be purified with 1 liter columns or protein A affinity chromatography, cation exchange chromatography, and anion exchange chromatography. The identity, purity, potency, safety, and impurities of the drug product will be evaluated by >15 test methods. The pharmacokinetics, immune response, safety pharmacology, and toxicology will be tested for the first time in adult Rhesus monkeys in an initial dose-ranging study. If successful, this research will provide the basis for a Pre-IND Meeting with the FDA, and entry of AGT-110 drug development into the phases of GLP pharmacology and GMP manufacturing. The overall goal of this work is the development of brain penetrating biologic TNFI, so that the pro-inflammatory effects of TNFalpha in neural disease can be suppressed.

描述（由申请人提供）：肿瘤坏死因子（TNF）-α在与衰老相关的神经退行性疾病（例如阿尔茨海默病或帕金森病）中发挥促炎作用。 TNF诱饵受体等生物TNF抑制剂（TNFI）不能用于治疗脑部疾病，因为TNFI是大分子，不能穿过血脑屏障（BBB）。目前的工作继续进行血脑屏障穿透性生物TNFI的药物开发，该TNFI是人II型TNF受体(TNFR)的重新设计形式，其中TNFR以IgG融合蛋白的形式产生。 IgG 部分是针对人胰岛素受体 (HIR) 的基因工程单克隆抗体 (MAb)。 HIRMAb-TNFR 融合蛋白被命名为 AGT-110。 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。在这个 II 期 SBIR 项目中，将开发大规模生产 HIRMAb-TNFR 融合蛋白的方法，适合生产临床试验批次的研究药物。将稳定转染的宿主细胞培养在50L生物反应器中，用1升柱或Protein A亲和层析、阳离子交换层析、阴离子交换层析纯化融合蛋白。药品的特性、纯度、效力、安全性和杂质将通过超过 15 种测试方法进行评估。在初始剂量范围研究中，将首次在成年恒河猴中测试药代动力学、免疫反应、安全药理学和毒理学。如果成功，这项研究将为与 FDA 举行 IND 前会议以及 AGT-110 药物开发进入 GLP 药理学和 GMP 制造阶段提供基础。这项工作的总体目标是开发脑穿透性生物TNFI，从而抑制TNFα在神经疾病中的促炎作用。