Novel therapeutic approaches to treatment of botulinum neurotoxin poisoning

治疗肉毒杆菌神经毒素中毒的新方法

基本信息

批准号：
8253685
负责人：
Konstantin Ichtchenko
金额：
$ 123.84万
依托单位：
NEW YORK UNIVERSITY SCHOOL OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-04-15 至 2016-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8253685
关键词：
Affect Amino Acid Sequence Antidotes Binding Binding Sites Biological Assay Bioterrorism Blood Circulation Bontoxilysin Botulinum Toxin Type A Botulism Breathing Bypass Caring Cells Centers for Disease Control and Prevention (U.S.)Cessation of life Clinical Coupled Cytoplasm Cytosol Data Development Disease Outbreaks Dose Drug Formulations Drug Kinetics Effectiveness Elements Endosomes Epithelial Exposure to Failure Food General Population Genetic Goals Hospitals Hour Human Immune Immunization Immunoglobulin Therapy Immunoglobulins In Vitro Ingestion Intestines Intoxication Intravenous Laboratories Lead Length Licensing Life Light Mechanical Ventilators Metalloproteases Methodology Methods Modality Modeling Molecular Motor Neurons Mus Neurons Oral Passive Immunotherapy Pharmacodynamics Physiological Poisoning Preparation Process Production Program Development Property Proteolysis Recombinants Recovery Research Respiratory Muscles Route S-nitro-N-acetylpenicillamine SNAP receptor Safety Serotyping Site Specificity System Testing Therapeutic Therapeutic Agents Time Toxic effect Toxicity Tests Toxin Treatment Efficacy Vaccination Vaccines Wound Infection base biodefense botulinum botulinum toxin type B design disorder prevention disulfide bond high risk in vivo inhibitor/antagonist intraperitoneal meetings neurotransmitter release novel novel therapeutic intervention novel therapeutics peptidomimetics pharmacokinetic characteristic pre-clinical protein aminoacid sequence response syntaxin therapeutic development therapeutic evaluation trafficking uptake

项目摘要

DESCRIPTION (provided by applicant): Human botulism can result from the ingestion of food contaminated with botulinum neurotoxin (BoNT), from the infection of wounds or intestine, or from direct inhalation of the toxin, which can be transcytosed across epithelial barriers as a physiologically active molecule. Intoxication, if untreated, may lead to death due to respiratory muscle failure, while recovery from botulism is a prolonged process (up to several months) and may require intensive and expensive hospital care. Due to their extreme toxicity and ease of production, BoNTs are classified as one of the six highest-risk threat agents for bioterrorism by the US Center for Disease Control and Prevention (CDC). Developing antidotes to botulinum neurotoxins is a priority goal of the Biodefense Research Agenda. Our laboratory has developed genetic constructs and expression systems based on recombinant, full- length, atoxic BoNT derivatives that preserve key structural features of the wt BoNT/A required for physiological trafficking, and enable the development of novel anti-BoNT therapeutics (ABTs) that will be effective in a post-exposure setting by oral or inhalational routes. The construction and expression of these atoxic BoNT derivatives provide molecular delivery vehicles that target motor neurons, and can contain specific sites for enzymatic attachment of a wide variety of cargo and tracker molecules. The genetic constructs are modular by design, and enable the facile production of diverse candidate ABTs for evaluation. In this application we will test two approaches for delivering therapeutic moieties to target neurons: a) For the ABT-1 approach, therapeutic moieties effective against all BoNT serotypes will be incorporated directly into the amino acid sequence of the BoNT atoxic light chain (LC) metalloprotease, which will be expressed in combination with the BoNT/A heavy chain (HC) as full-length ABT-1; b) For the ABT-2 approach, a therapeutic moiety specifically active against wt BoNT/A will be coupled to the full length ABT-2 by site- selective enzymatic attachment to the ABT-2 precursor, carried by the BoNT/A heavy chain. The full-length BoNT derivatives we propose are expected to have unique advantages with respect to their ability to compete with wild type toxin for neuronal binding sites, and to mimic the endosome-to-cytosol channeling required for delivery of ABTs to the neuronal cytosol. Our current methods can produce sufficient quantities for in vitro and in vivo testing. These ABTs can be administered in a post-exposure setting by oral or inhalational routes.

描述（由申请人提供）：人类肉毒杆菌症可能是由于摄入被肉毒杆菌神经毒素（BONT）污染的食物，受伤或肠的感染或直接吸入毒素的食物，这些食物可以跨表皮屏障作为物理活性分子跨性别。如果未经治疗，中毒可能会导致呼吸道肌肉衰竭导致死亡，而从肉毒杆菌中恢复是一个长时间的过程（长达几个月），可能需要大量且昂贵的医院护理。由于它们的极端毒性和易于生产，因此BONT被归类为美国疾病控制与预防中心（CDC）的六种最高风险威胁药物之一。开发对肉毒肉毒神经毒素的解毒剂是生物形式研究议程的优先目标。我们的实验室已经开发了基于重组，全长，毒性的BONT衍生物的遗传构建体和表达系统，这些衍生物保留了生理运输所必需的WT BONT/A的关键结构特征，并能够开发新型的抗BONT治疗学（ABT），这些抗Bont疗法（ABT）将在口头或毛线途径后有效地暴露于后的情况下有效。这些毒性BONT衍生物的构造和表达提供了靶向运动神经元的分子递送车，并且可以包含特定位点，以酶促连接各种货物和跟踪器分子。遗传构建体是通过设计模块化的，可以使各种候选ABT的便利产生进行评估。在此应用中，我们将测试两种用于将治疗性部分传递到靶向神经元的方法： b）对于ABT-2方法，针对WT BONT/A专门活跃的治疗部分将通过位点选择性酶促连接到ABT-2前体，由BONT/A重链携带。我们提出的全长BONT衍生物有望具有与野生型毒素在神经元结合位点竞争的能力，并模仿abt递送到神经元细胞质所需的内体到胞质溶液。我们当前的方法可以产生足够的体外和体内测试。这些ABT可以通过口服或吸入路线在暴露后设置中给药。