Aerosol Delivery of a Recombinant Butyrylcholinesterase "BioShield" to Protect Ag

气溶胶递送重组丁酰胆碱酯酶“BioShield”以保护 Ag

基本信息

批准号：
7612606
负责人：
Yvonne J Rosenberg
金额：
$ 28.8万
依托单位：
PLANTVAX, INC.
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-07-08 至 2012-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7612606
关键词：
Ache Acute Aerosol Drug Therapy Aerosols Agarose Chromatography Agriculture Animals Antibodies Antidotes Area Atropine Award Binding Biological Availability Biomanufacturing Bioshield Biotechnology Blood Blood Circulation Breathing Businesses Butyrylcholinesterase Cell Line Cells Cessation of life Chemicals Chlorpyrifos Chronic Deposition Development Devices Diazinon Disadvantaged Dose Drug Kinetics Enzymes Ethics Exhibits Feasibility Studies Functional disorder Genes Glycoproteins Grant Human In Situ Individual Industry Inhalation Exposure Injection of therapeutic agent Insecticides Institutes Lethal Dose 50 Liquid substance Lung Macaca Maryland Measures Mediating Medicine Modeling Monkeys Morbidity - disease rate Mus National Institute of Neurological Disorders and Stroke Neuromuscular Junction Neuropathy Neurotoxins Organophosphates Organophosphorus Compounds Oximes Paraoxon Pattern Pharmaceutical Preparations Phase Physiological Plasma Poison Poisoning Polysaccharides Powder dose form Procainamide Process Production Proteins Protocols documentation Public Health Radionuclide Imaging Recombinant Proteins Recombinants Route Safety Sampling Scanning Sepharose Serum Structure of parenchyma of lung Supportive care Symptoms System Technetium 99m Testing Therapeutic Time Toxic effect Translational Research Treatment Protocols Universities Veterinarians Work Writing aerosolized bioscavenger carbamate insecticide cell bank cholinergic cholinergic synapse efficacy testing experience expression cloning immunogenic immunogenicity innovation killings meetings neuromuscular neurotoxic neurotoxicity nonhuman primate organophosphate poisoning pesticide exposure prevent public health relevance respiratory response safety study safety testing stoichiometry therapeutic protein user-friendly

项目摘要

DESCRIPTION (provided by applicant): Organophosphorus (OPs) compound are potent neurotoxic chemicals that are widely used in medicine, industry and agriculture; most notably as insecticides. The neurotoxicity which is primarily a result of AChE inhibition, may take the form of cholinergic crisis and death as a consequence of acute exposure or psychiatric symptoms and delayed neuropathy following chronic exposure. Usually, treatment of insecticide poisoning consists of supportive care and specific therapy e.g. atropine and oximes, which often fail to prevent morbidity or death. Currently, recombinant (r) BChE is a leading pre-exposure treatment candidate for OP toxicity due to its potent bioscavenging ability but because of its 1:1 stoichiometry with OP, large doses will be required if delivered by i.m. or subcutaneously routes. In order to reduce the dose required for systemic approaches for delivering BChE, we plan to create a protective "BChE bioshield" of aerosolized tetrameric rBChE into the lungs to detoxify incoming (inhaled) OPs in situ, thus preventing the OP's entry into the systemic circulation and avoiding the respiratoty usually associated with inhalation exposure. Phase I represents a feasibility study in mice to examine (i) the patterns of deposition of either liquid or powdered PEG-rMaBChE radioaerasols delivered to the lungs by Microsprayer or insufflator respectively (ii) the persistence of the "bioshield" and (iii) the degree of protection it affords against 50% LD50 of the liquid aerosolized OP insecticide paraoxon as measured by percent inhibition of RBC AChE and serum BChE. In Phase II, the efficacy of the aerosolized PEG-rMaBChE to protect against OP toxicity will be tested in a homologous macaque model. A homologous system, which does not make anti-BChE antibodies, is critical for accurate efficacy, pharmacokinetics and safety testing of a recombinant therapeutic protein which may be required as a multiple administrations. During this time, Master and Working Cell Banks of CHO-K1 producing rHuBChE will be generated and process development begun on the production, purification and PEG-ylation protocols. The efficacy of an aerasolized PEG-rHuBChE "bioshield" to prevent paraoxon toxicity will be tested in macaques and compared with the homologous rMaBChE enzyme. It is anticipated that aerosol delivery will reduce the dose required for protection because of its concentration in the same areas as the inhaled insecticide. The development of an innovative "Microsprayer" type device would permit a user friendly treatment to be delivered before the known use/release of insecticides. Public Health Relevance: Many glycoproteins which are potent antidotes, exhibit very important physiological functions but cannot be used as therapeutic treatments because they are quickly removed from the circulation or their ability to protect requires large doses if given i.m. The aim of this project is to butyrylcholinesterase as an aerosol in a pulmonary delivery device to that it deposits in the lung and forms a "bioshield" and detoxifies inhaled indecticides poisons in the lung and prevernts them from reaching the blood and the neuromuscular junctions where they can quickly cause severe toxicity.

描述（由申请人提供）：有机磷（OPs）化合物是强效神经毒性化学物质，广泛应用于医药、工业和农业；最引人注目的是作为杀虫剂。主要由乙酰胆碱酯酶抑制引起的神经毒性，可能会因急性接触或慢性接触后出现精神症状和迟发性神经病变而导致胆碱能危象和死亡。通常，杀虫剂中毒的治疗包括支持治疗和特定治疗，例如药物治疗。阿托品和肟通常无法预防发病或死亡。目前，重组 (r) BChE 因其强大的生物清除能力而成为 OP 毒性的主要暴露前治疗候选者，但由于其与 OP 的化学计量为 1:1，如果通过肌内递送，则需要大剂量。或皮下途径。为了减少全身方法输送 BChE 所需的剂量，我们计划创建一个保护性“BChE 生物盾”，将雾化四聚体 rBChE 送入肺部，以在原位对传入（吸入）的 OP 进行解毒，从而防止 OP 进入体循环并避免通常与吸入暴露相关的呼吸。第一阶段代表在小鼠中进行的可行性研究，以检查（i）分别通过微喷雾器或吹入器输送到肺部的液体或粉末状 PEG-rMaBChE 放射性气溶胶的沉积模式（ii）“生物防护罩”的持久性和（iii）根据 RBC AChE 和血清 BChE 的抑制百分比来测量，它对液体雾化 OP 杀虫剂对氧磷的 LD50 为 50%。在第二阶段，将在同源猕猴模型中测试雾化 PEG-rMaBChE 预防 OP 毒性的功效。不产生抗 BChE 抗体的同源系统对于重组治疗性蛋白的准确功效、药代动力学和安全性测试至关重要，而重组治疗性蛋白可能需要多次给药。在此期间，将生成生产 rHuBChE 的 CHO-K1 主细胞库和工作细胞库，并开始生产、纯化和 PEG 化方案的工艺开发。雾化 PEG-rHuBChE“生物盾”预防对氧磷毒性的功效将在猕猴中进行测试，并与同源 rMaBChE 酶进行比较。预计气雾剂输送将减少保护所需的剂量，因为其浓度与吸入杀虫剂相同。创新的“微喷雾器”型装置的开发将允许在已知的使用/释放杀虫剂之前提供用户友好的处理。公共健康相关性：许多糖蛋白是有效的解毒剂，表现出非常重要的生理功能，但不能用作治疗方法，因为它们会很快从循环中清除，或者如果肌内注射，它们的保护能力需要大剂量。该项目的目的是将丁酰胆碱酯酶作为肺部输送装置中的气雾剂，沉积在肺部并形成“生物盾”，对肺部吸入的杀虫剂毒物进行解毒，并防止它们到达血液和神经肌肉接头。能很快引起严重的毒性。