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: 8246989
• 负责人: RUBEN J. BOADO
• 金额: $ 50.81万
• 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/8246989
• 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; Public Health; 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; public health relevance; 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（由申请人提供）：单克隆抗体（MAb）是许多脑疾病的潜在新治疗剂，包括阿尔茨海默病（AD）、帕金森病、疯牛病、西尼罗河脑炎、神经- AIDS、脑损伤、脑癌或多发性硬化.在几乎所有情况下，给予血液的单克隆抗体治疗剂必须能够进入脑内的靶位点。然而，MAb是不穿过血脑屏障（BBB）的大分子药物。血脑屏障问题阻碍了抗体药物的脑药物开发。这项研究将开发一种新的抗体药物递送技术，该技术也可用于其他器官，并将应用于AD。这项工作是基于由2种抗体组成的融合蛋白的基因工程。一种抗体是针对AD的A β淀粉样肽的治疗性抗体，另一种抗体是药物递送系统，其针对人BBB上的内源性转运蛋白。I期研究完成了以下内容：（1）表达异源四聚体融合蛋白的串联载体的基因工程，（2）克隆在无血清培养基中表达高水平融合抗体的永久转染的宿主细胞系，（3）融合抗体的生物化学和功能表征，和（4）测定所述融合蛋白在成年恒河猴中的血浆药代动力学（PK）和脑摄取。II期研究将完成以下工作：（1）宿主细胞系在50 L生物反应器中生长，然后进行可在GMP实验室中重复的3柱下游处理;（2）融合蛋白的生化分析（超过15项分析试验）;（3）恒河猴剂量确定PK和毒性研究。这些研究将使未来能够提交IND，用于对这种新的AD融合蛋白进行人体试验。 公共卫生相关性：单克隆抗体是生物技术的强大的新治疗产品。抗体药物可以应用于许多严重的脑部疾病，如阿尔茨海默病（AD）、帕金森病、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，不能开发用于这些疾病的抗体药物，因为抗体药物不能穿过血脑屏障（BBB）。目前的研究将开发一种用于大脑的抗体药物递送的新技术，该技术可应用于AD等疾病。