Bioengineering of a New Antibody Drug Delivery Technology

新型抗体药物递送技术的生物工程

• 批准号: 8055209
• 负责人: RUBEN J. BOADO
• 金额: $ 33.48万
• 依托单位: ARMAGEN TECHNOLOGIES, INC.
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-15 至 2013-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8055209
• 关键词: Accounting; Acquired Immunodeficiency Syndrome; Active Immunization; Adult; Affinity; Affinity Chromatography; Alzheimer' s Disease; Amyloid beta-Protein; Animals; Anions; Antibodies; Appearance; Autopsy; Binding; Biochemical; Biological Assay; Biomedical Engineering; Bioreactors; Biotechnology; Blood; Blood - brain barrier anatomy; Body Weight; Bovine Spongiform Encephalopathy; Brain; Brain Diseases; Brain Injuries; Carbohydrates; Cations; Cell Line; Cells; Cerebrum; Chimeric Proteins; Chinese Hamster; Chinese Hamster Ovary Cell; Chromatography; Clinical Chemistry Tests; Cloning; Complex; DNA; Dementia; Development; Disease; Dose; Drug Delivery Systems; Drug Kinetics; Encephalitis; Endotoxins; Engineering; Enzyme-Linked Immunosorbent Assay; Exclusion; Filtration; Fluorescence Microscopy; Future; Genetic Engineering; Glial Fibrillary Acidic Protein; Growth; Hematoxylin and Eosin Staining Method; Hemorrhage; High Pressure Liquid Chromatography; Histocytochemistry; Human; Immunoglobulin G; Immunotherapy; Injection of therapeutic agent; Insulin Receptor; Isoelectric Focusing; Macaca mulatta; Malignant neoplasm of brain; Mediating; Monoclonal Antibodies; Multiple Sclerosis; Organ; Organ Weight; Ovary; Parkinson Disease; Passive Immunization; Peptide Mapping; Peptides; Perfusion; Peripheral; Pharmaceutical Preparations; Phase; Plasma; Primates; Process; Production; Progress Reports; Protein Binding; Proteins; Prussian blue; Publications; Recombinant Fusion Proteins; Reference Standards; Relative (related person); Reporting; Research; Senile Plaques; Serum-Free Culture Media; Site; Small Business Innovation Research Grant; Symptoms; Technology; Temperature; Testing; Therapeutic Monoclonal Antibodies; Therapeutic antibodies; Time; Tissue Stains; Toxic effect; Toxicology; Transgenic Mice; Urine; West Nile virus; Western Blotting; Work; amyloid peptide; base; brain tissue; drug development; fluoro jade; human INSR protein; immunocytochemistry; manufacturing process; molecular trojan horse; nano; neonatal Fc receptor; neuropathology; new technology; novel therapeutics; phase 1 study; phase 2 study; prevent; programs; receptor; receptor binding; uptake; vector

DESCRIPTION (provided by applicant): Monoclonal antibodies (MAb) are potential new therapeutics for many brain diseases, including Alzheimer's disease (AD), Parkinson's disease, mad cow disease, West Nile encephalitis, neuro- AIDS, brain injury, brain cancer, or multiple sclerosis. In almost all cases, it is necessary that the MAb therapeutic that is administered into the blood be able to access target sites within the brain. However, MAb's are large molecule drugs that do not cross the blood-brain barrier (BBB). The BBB problem prevents the brain drug development of antibody drugs. The proposed research will develop a new technology for antibody drug delivery to brain, which could also be used for other organs, and the new technology will be applied to AD. This work is based on the genetic engineering of a fusion protein comprised of 2 antibodies. One antibody is the therapeutic antibody against the Abeta amyloid peptide of AD, and the other antibody is a drug delivery system, which is directed at an endogenous transporter on the human BBB. The Phase I studies accomplished the following: (1) genetic engineering of a tandem vector expressing the hetero-tetrameric fusion protein, (2) cloning of a permanently transfected host cell line that expresses high levels of the fusion antibody in serum free medium, (3) biochemical and functional characterizion of the fusion antibody, and (4) determination of the plasma pharmacokinetics (PK) and brain uptake of the fusion protein in the adult Rhesus monkey. The phase II studies will accomplish the following: (1) growth of the host cell line in a 50L bioreactor, followed by 3-column downstream processing that can be replicated in a GMP lab; (2) biochemical analysis of the fusion protein with over 15 analytical tests; (3) dose finding PK and toxicity study in Rhesus monkeys. These studies will enable future submission of an IND for human testing of this new fusion protein for AD. PUBLIC HEALTH RELEVANCE: Monoclonal antibodies are powerful new therapeutic products of biotechnology. Antibody drugs could be applied to many serious brain disorders, such as Alzheimer's disease (AD), Parkinson's disease, mad cow disease, West Nile encephalitis, neuro-AIDS, brain injury, brain cancer, or multiple sclerosis. However, antibody drugs cannot be developed for these disorders, because the antibody drugs do not cross the blood-brain barrier (BBB). The present research will develop a new technology for the drug delivery of antibody drugs for the brain, which could be applied to diseases such as AD.

描述(申请人提供)：单抗(MAb)是许多脑部疾病的潜在新疗法，包括阿尔茨海默病(AD)、帕金森氏病、疯牛病、西尼罗河脑炎、神经艾滋病、脑损伤、脑癌或多发性硬化症。在几乎所有的情况下，注射到血液中的单抗治疗药物都必须能够进入大脑中的靶点。然而，单抗是不能通过血脑屏障(BBB)的大分子药物。血脑屏障问题阻碍了脑部药物抗体药物的开发。这项拟议的研究将开发一种将抗体药物输送到大脑的新技术，该技术也可以用于其他器官，新技术将应用于AD。这项工作是基于一种由两种抗体组成的融合蛋白的基因工程。一种抗体是针对AD的Aβ淀粉样多肽的治疗性抗体，另一种抗体是针对人血脑屏障上的内源性转运蛋白的药物递送系统。第一阶段的研究完成了以下工作：(1)表达异源四聚体融合蛋白的串联载体的基因工程；(2)在无血清介质中高水平表达融合抗体的永久性转基因宿主细胞系的克隆；(3)融合抗体的生化和功能特性；(4)成年恒河猴对融合蛋白的血浆药代动力学(PK)和脑摄取的测定。第二阶段的研究将完成以下工作：(1)宿主细胞系在50L生物反应器中的生长，随后是可在GMP实验室复制的3柱下游加工；(2)融合蛋白的生化分析，包括15项以上的分析测试；(3)恒河猴的剂量发现PK和毒性研究。这些研究将使今后提交IND以供人类测试这种新的AD融合蛋白成为可能。 与公共卫生相关：单抗是生物技术强有力的新治疗产品。抗体药物可用于许多严重的大脑疾病，如阿尔茨海默病(AD)、帕金森氏病、疯牛病、西尼罗河脑炎、神经艾滋病、脑损伤、脑癌或多发性硬化症。然而，抗体药物无法针对这些疾病开发，因为抗体药物不能跨越血脑屏障(BBB)。目前的研究将开发一种新的脑部抗体药物给药技术，可应用于AD等疾病。