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Neurotrophin Drug Development for Parkinson's Disease

帕金森病的神经营养蛋白药物开发

基本信息

批准号：
7877345
负责人：
William M Pardridge
金额：
$ 33.69万
依托单位：
UNIVERSITY OF CALIFORNIA LOS ANGELES
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-02-01 至 2013-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7877345
关键词：
Affect Affinity Affinity Chromatography Age-Months Amphetamines Antibodies Apomorphine Area Behavior Biochemical Biological Assay Bioreactors Blood Blood - brain barrier anatomy Blood capillaries Brain Brain Part Brain region C57BL/6 Mouse COS Cells Cations Cell Line Cells Cephalic Cerebellum Chemistry Chimeric Proteins Chinese Hamster Chinese Hamster Ovary Cell Chromatography Chronic Clinical Clinical Pharmacology Cloning Complementary DNA Control Groups Corpus striatum structure Cytomegalovirus DNA Data Degenerative Disorder Development Dihydrofolate Reductase Dose Drug Delivery Systems Drug Kinetics Engineering Enzyme-Linked Immunosorbent Assay Equus caballus Euthanasia Experimental Parkinsonism Feasibility Studies Female Filtration G-substrate Genes Genetic Engineering Hepatitis B Virus Histology Human Hybridomas IgG1 Immunoglobulin G Immunoglobulin Variable Region Inbred BALB C Mice Insulin Receptor Introns Label Laboratories Lesion Light Macaca mulatta Measurement Measures Mediating Methods Methotrexate Monoclonal Antibodies Mus Nerve Degeneration Nerve Growth Factor Receptors Neurotoxins Organ Organ Weight Ovary Oxidopamine Parkinson Disease Patients Pharmaceutical Preparations Pharmacodynamics Plasma Proteins Public Health RNA Splicing Radio Rattus Reaction Time Research Rodent Rotation Series Serum Serum-Free Culture Media Simian virus 40 System Testing Therapeutic Therapeutic Index Toxic effect Toxin Transferrin Transferrin Receptor Tyrosine 3-Monooxygenase Western Blotting analog behavior measurement behavior test bovine growth hormone brain cell capillary drug development drug discovery drug efficacy enzyme activity fusion gene gene cloning human INSR protein immunocytochemistry immunogenicity in vivo light microscopy male molecular trojan horse mouse model nervous system disorder neurotrophic factor novel novel therapeutics plasmid DNA pre-clinical promoter public health relevance receptor receptor binding research study response sex uptake

项目摘要

DESCRIPTION (provided by applicant): Parkinson's disease (PD) affects 1 million people in the U.S., and is a degenerative disease caused by the loss of brain cells in the nigral-striatal tract. The most potent protective factor for this area of the brain is the neurotrophin, glial derived neurotrophic factor (GDNF). However, GDNF drug development in PD has failed, because the neurotrophin does not cross the blood-brain barrier (BBB). Consequently, the GDNF was administered to patients with PD via a trans-cranial drug delivery system that does not effectively deliver the drug to the part of the brain involved in PD. The present research will genetically engineer a new form of GDNF that is enabled to cross the BBB via receptor-mediated transport. The PI has previously genetically engineered a chimeric MAb against the mouse transferrin receptor (TfR), designated cTfRMAb, that crosses the BBB in the blood-to-brain direction via receptor-mediated transport on the mouse BBB TfR. In addition, the PI has genetically engineered, and expressed a fusion protein of GDNF and the cTfRMAb, which is designated the cTfRMAb-GDNF fusion protein. The bi-functionality of the fusion protein was verified in transient expression experiments. The present research will produce a permanently transfected host cell line using Chinese hamster ovary (CHO) cells that secrete the cTfRMAb-GDNF fusion protein in high amounts so that 200 mg of the fusion protein can be produced. This fusion protein will then be used for pharmacokinetics studies in the mouse, for time response and dose response efficacy studies in an experimental mouse model of PD using C57Bl/6 mice lesioned with intra-cranial 6- hydroxydopmaine. In addition a toxicity study will be performed where male and female C57Bl/6 mice are treated chronically with the cTfRMAb-GDNF fusion protein; the histology of brain and other major organs will then be examined. The formation of anti-fusion protein antibodies will be examined using a novel sandwich ELISA. The drug efficacy in experimental PD will be validated with rotation behavior measurements, and assays of striatal tyrosine hydroxylase. This research will provide the necessary pre-clinical pharmacology to support an IND filing for the treatment of humans with PD using genetically engineered forms of GDNF that cross the BBB via receptor-mediated transport. PUBLIC HEALTH RELEVANCE: Parkinson's disease (PD) affects 1 million people in the U.S. The most potent form of treatment of PD is a neurotrophin called GDNF; however, GDNF does not cross the blood-brain barrier (BBB). The present research will test in experimental PD in mice the efficacy of a new form of GDNF, wherein the neurotrophin is re-engineered as a fusion protein with a monoclonal antibody that crosses the BBB via receptor-mediated transport.

描述（由申请人提供）：帕金森病（PD）影响美国100万人，并且是由黑质-纹状体束中的脑细胞损失引起的退行性疾病。对大脑的这个区域最有效的保护因子是神经营养因子，胶质源性神经营养因子（GDNF）。然而，GDNF在PD中的药物开发已经失败，因为神经营养因子不能穿过血脑屏障（BBB）。因此，GDNF通过经颅药物递送系统施用于患有PD的患者，该系统不能有效地将药物递送到参与PD的大脑部分。目前的研究将通过基因工程改造一种新形式的GDNF，使其能够通过受体介导的运输穿过BBB。PI先前已对针对小鼠转铁蛋白受体（TfR）的嵌合MAb（命名为cTfRMAb）进行了基因工程改造，该嵌合MAb通过小鼠BBB TfR上的受体介导的转运以血液至脑的方向穿过BBB。此外，PI已经遗传工程化，并表达GDNF和cTfRMAb的融合蛋白，其被指定为cTfRMAb-GDNF融合蛋白。在瞬时表达实验中验证了融合蛋白的双功能性。本研究将使用中国仓鼠卵巢（CHO）细胞产生永久转染的宿主细胞系，所述细胞以高量分泌cTfRMAb-GDNF融合蛋白，使得可以产生200 mg融合蛋白。然后将该融合蛋白用于小鼠中的药代动力学研究，用于使用颅内6-羟基多巴胺损伤的C57 B1/6小鼠的PD实验小鼠模型中的时间响应和剂量响应功效研究。此外，将进行毒性研究，其中雄性和雌性C57 B1/6小鼠用cTfRMAb-GDNF融合蛋白长期治疗;然后将检查脑和其他主要器官的组织学。将使用新型夹心ELISA检查抗融合蛋白抗体的形成。将通过旋转行为测量和纹状体酪氨酸羟化酶测定来验证实验性PD中的药物疗效。这项研究将提供必要的临床前药理学，以支持IND申请，用于使用通过受体介导的转运穿过BBB的GDNF基因工程形式治疗PD患者。公共卫生相关性：帕金森氏病（PD）影响着美国100万人。PD最有效的治疗形式是一种称为GDNF的神经营养因子;然而，GDNF不会穿过血脑屏障（BBB）。本研究将在小鼠实验性PD中测试GDNF的新形式的功效，其中神经营养因子被重新设计为与单克隆抗体的融合蛋白，其通过受体介导的转运穿过BBB。