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：
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
项目类别：
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&apos 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

项目摘要

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在与衰老有关的神经退行性疾病中起促炎作用，例如阿尔茨海默氏病或帕金森氏病。生物TNF抑制剂（TNFI），例如TNF诱饵受体，无法用于脑部疾病，因为TNFI是不跨越血脑屏障（BBB）的大分子。目前的工作延续了BBB渗透生物TNFI的药物发展，这是人类II型TNF受体（TNFR）的重新设计形式，其中TNFR作为IgG融合蛋白生产。 IgG部分是针对人类胰岛素受体（HIR）的基因工程单克隆抗体（MAB）。 Hirmab-TNFR融合蛋白称为AGT-1110。 Hirmab-TNFR融合蛋白的Hirmab部分充当分子木马马，可通过内源性BBB胰岛素受体上的受体介导的转运在BBB穿越熔融的诱饵受体。在此II阶段SBIR项目中，将开发出适合生产临床试验的大规模生产Hirmab-TNFR融合蛋白的方法。稳定转染的宿主细胞将在50升生物反应器中培养，融合蛋白将用1升色谱柱或A蛋白A亲和色谱，阳离子交换色谱和阴离子交换色谱纯化。药物的身份，纯度，效能，安全性和杂质将通过> 15种测试方法进行评估。在最初的剂量范围研究中，将首次在成年恒河猴中首次对药代动力学，免疫反应，安全药理学和毒理学进行测试。如果成功，这项研究将为与FDA的预先开会，并将AGT-1110药物开发进入GLP药理学和GMP制造阶段的基础。这项工作的总体目的是发展脑穿透生物学TNFI，因此可以抑制TNFalpha对神经疾病的促炎作用。