Neurotrophin Drug Development for Parkinson's Disease

帕金森病的神经营养蛋白药物开发

• 批准号: 8214521
• 负责人: William M Pardridge
• 金额: $ 33.01万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-02-01 至 2013-06-27
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8214521
• 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; Body Weight; 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; GDNF receptors; Genes; Genetic Engineering; Hepatitis B Virus; Histology; Human; Hybridomas; IgG1; Immunoglobulin G; Immunoglobulin Variable Region; Inbred BALB C Mice; Insulin Receptor; Introns; Label; 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; The Jackson Laboratory; 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

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）：帕金森氏病（PD）影响了美国的100万人，这是由于ni骨纹状体中脑细胞丧失引起的一种退化性疾病。大脑该区域的最有效的保护因子是神经营养蛋白，神经营养衍生的神经营养因子（GDNF）。然而，PD中的GDNF药物开发失败了，因为神经营养蛋白没有跨越血脑屏障（BBB）。因此，通过跨颅药物输送系统对PD患者进行了GDNF，该药物没有有效地将药物输送到参与PD的大脑部分。本研究将在基因上设计一种新形式的GDNF，该形式可以通过受体介导的运输跨越BBB。 PI先前已经针对指定的CTFRMAB的小鼠转铁蛋白受体（TFR）进行了基因设计的嵌合MAB，该受体通过受体介导的小鼠BBB BBB TFR在血液到脑方向上穿越BBB。此外，PI具有基因设计，并表达了GDNF和CTFRMAB的融合蛋白，该蛋白被指定为CTFRMAB-GDNF融合蛋白。在瞬态表达实验中验证了融合蛋白的双功能。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，该细胞分泌CTFRMAB-GDNF融合蛋白，以便产生200 mg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，用于使用C57BL/6小鼠在PD的实验小鼠模型中进行时间响应和剂量反应疗效研究，该模型使用颅内6-羟基羟基脑的病变。另外，将在雄性和雌性C57BL/6小鼠用CTFRMAB-GDNF融合蛋白长期治疗的情况下进行毒性研究。然后将检查大脑和其他主要器官的组织学。将使用新型的三明治ELISA检查抗融合蛋白抗体的形成。实验PD中的药物功效将通过旋转行为测量和纹状体酪氨酸羟化酶的测定验证。这项研究将提供必要的临床前药理学，以使用基因工程形式的GDNF的GDNF进行IND申请，以通过受体介导的转运跨越BBB。 公共卫生相关性：帕金森氏病（PD）在美国影响100万人，最有效的PD治疗形式是一种称为GDNF的神经营养素；但是，GDNF不会越过血脑屏障（BBB）。本研究将在小鼠实验性PD中测试一种新形式的GDNF的功效，其中神经营养蛋白被重新设计为具有单克隆抗体的融合蛋白，该抗体通过受体介导的转运穿越BBB。

项目成果

Brain penetrating IgG-erythropoietin fusion protein is neuroprotective following intravenous treatment in Parkinson's disease in the mouse.

脑穿透性 IgG-促红细胞生成素融合蛋白对小鼠帕金森病进行静脉注射治疗后具有神经保护作用。

• DOI: 10.1016/j.brainres.2011.01.061
• 发表时间: 2011
• 期刊: Brain research
• 影响因子: 2.9
• 作者: Zhou,Qing-Hui;Hui,EricKa-Wai;Lu,JeffZhiqiang;Boado,RubenJ;Pardridge,WilliamM
• 通讯作者: Pardridge,WilliamM

Neuroprotection with a brain-penetrating biologic tumor necrosis factor inhibitor.

使用脑穿透性生物肿瘤坏死因子抑制剂进行神经保护。

• DOI: 10.1124/jpet.111.185876
• 发表时间: 2011
• 期刊: The Journal of pharmacology and experimental therapeutics
• 作者: Zhou,Qing-Hui;Sumbria,Rachita;Hui,EricKa-Wai;Lu,JeffZhiqiang;Boado,RubenJ;Pardridge,WilliamM
• 通讯作者: Pardridge,WilliamM

Intravenous treatment of experimental Parkinson's disease in the mouse with an IgG-GDNF fusion protein that penetrates the blood-brain barrier.

• DOI: 10.1016/j.brainres.2010.06.059
• 发表时间: 2010-09-17
• 期刊: Brain research
• 影响因子: 2.9
• 作者: Fu A;Zhou QH;Hui EK;Lu JZ;Boado RJ;Pardridge WM
• 通讯作者: Pardridge WM