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) 在美国影响着 100 万人，是一种由黑质-纹状体束中脑细胞损失引起的退行性疾病。大脑该区域最有效的保护因子是神经营养蛋白，即神经胶质源性神经营养因子（GDNF）。然而，PD 中的 GDNF 药物开发失败了，因为神经营养蛋白不能穿过血脑屏障 (BBB)。因此，GDNF 通过经颅药物输送系统给予 PD 患者，但该系统不能有效地将药物输送到与 PD 相关的大脑部分。目前的研究将对 GDNF 进行基因改造，使其能够通过受体介导的转运穿过 BBB。 PI 此前已通过基因工程改造了一种针对小鼠转铁蛋白受体 (TfR) 的嵌合 MAb，命名为 cTfRMAb，该单克隆抗体通过小鼠 BBB TfR 上受体介导的转运以血脑方向穿过 BBB。此外，PI经过基因工程改造，表达了GDNF和cTfRMAb的融合蛋白，称为cTfRMAb-GDNF融合蛋白。融合蛋白的双功能在瞬时表达实验中得到验证。目前的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，该细胞大量分泌cTfRMAb-GDNF融合蛋白，从而可以生产200毫克的融合蛋白。然后，该融合蛋白将用于小鼠的药代动力学研究，以及使用颅内 6-羟基多巴因损伤的 C57Bl/6 小鼠在 PD 实验小鼠模型中进行时间反应和剂量反应功效研究。此外，还将进行一项毒性研究，其中雄性和雌性 C57Bl/6 小鼠长期接受 cTfRMAb-GDNF 融合蛋白治疗；然后将检查大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。实验性 PD 中的药物疗效将通过旋转行为测量和纹状体酪氨酸羟化酶测定来验证。这项研究将提供必要的临床前药理学，以支持使用基因工程形式的 GDNF（通过受体介导的转运穿过 BBB）治疗人类 PD 的 IND 申请。公共健康相关性：帕金森病 (PD) 在美国影响着 100 万人。帕金森病最有效的治疗方法是一种称为 GDNF 的神经营养蛋白；然而，GDNF 不能穿过血脑屏障 (BBB)。目前的研究将在小鼠实验性 PD 中测试新型 GDNF 的功效，其中神经营养素被重新设计为与单克隆抗体的融合蛋白，通过受体介导的转运穿过 BBB。