RNA aptamers as novel antidotes against botulism

RNA适体作为肉毒中毒的新型解毒剂

• 批准号: 7388510
• 负责人: SHUOWEI CAI
• 金额: $ 20.44万
• 依托单位: UNIVERSITY OF MASSACHUSETTS DARTMOUTH
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-12-15 至 2009-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7388510
• 关键词: Adverse effects; Affinity; Antidotes; Antitoxins; Binding; Biological; Biological Assay; Bioterrorism; Blood Circulation; Bontoxilysin; Botulinum Toxin Type A; Botulism; Cells; Characteristics; Chemicals; Class; Classification; Communities; Diagnostic; Disease; Endopeptidases; Ensure; Environmental air flow; Equus caballus; Evolution; Gangliosides; Generations; Goals; Health; In Vitro; Intensive Care; Lead; Libraries; Ligands; Methods; National Institute of Allergy and Infectious Disease; Neuroblastoma; Neurons; Norepinephrine; Nucleic Acids; Oligonucleotides; PC12 Cells; Paralysed; Patients; Pharmaceutical Preparations; Poison; Principal Investigator; Prophylactic treatment; Proteins; Public Health; RNA; RNA library; Range; Reporting; Research; Research Project Grants; Risk; Security; Serotyping; Specificity; Symptoms; Therapeutic; Therapeutic Agents; Toxic effect; Toxin; Validation; aptamer; base; biodefense; combinatorial; cost; design; immunogenicity; man; novel; pathogen; programs; prophylactic; receptor; small molecule

DESCRIPTION (provided by applicant): Botulinum neurotoxins (BoNTs) are the most toxic substances known to man, and cause the deadly neuroparalytic disease, botulism. The flaccid paralysis caused by botulism lasts for months. BoNTs can be used as potential bioterrorism agents, and therefore, pose great threat to homeland security and public health. Currently no effective and safe antidote exists for prophylactic and therapeutic treatment for botulism. The only approved antidotes are equine antitoxins, which can only neutralize the toxin in the blood circulation, and have a very short treatment window. In addition, the equine antitoxins can cause severe side effects, excluding them as effective prophylactics. The only alternative for patients remains to be put on artificial ventilation under intensive care for several months. In this R21 exploratory research grant, we propose a new generation of antidotes against botulism, aptamers. Aptamers are unique oligonucleotides that have high affinity for their targets, ranging from small molecules to proteins. Aptamers have low toxicity, no reported immunogenicity, low cost to manufacture, and excellent storage stability. Those characteristics make aptamers ideal as antidotes against botulism. The goal of this research is to develop aptamers as antidotes against botulism. The proposed antidotes have potential to be both effective prophylactic treatment, and effective therapeutic drugs to treat botulism patients with symptoms. The specific aims to be pursued in the next two years are: (1) Selection and characterization of aptamers against BoNT. We will use SELEX to screen random RNA libraries, isolate the aptamers against BoNT, and prioritize aptamers based on binding affinity to BoNT. (2) Validation of anti-BoNT activity of aptamers. We will determine the inhibitory effects on both endopeptidase activity and toxin binding to its receptors, using both in vitro and cell-based assays. The aptamers identified from this study, therefore, will be used as lead compounds for the designing of more potent antidotes against botulism. This study will lead to a new class of antidotes against botulism. The research approach can be readily applicable to all seven serotypes of BoNT and other priority pathogens on biodefense agenda. In addition, the research proposed here will have significant impact on the rapid diagnostics of biodefense agents. The ever increasing health risks due to bioterrorism worldwide have placed a large burden on the scientific community to rapidly develop safe prophylactic and therapeutic treatments for biological and chemical toxins. This task has been a high priority for public health. There is lack of therapeutic and prophylactic drugs for botulism and most of the other biological toxins (as listed in NIAID priority pathogen list). Our project will result in aptamer-based drug candidates against botulism, a new class of antidotes against botulism, with many advantages over currently available antitoxin.

描述（由申请人提供）：肉毒神经毒素（BoNT）是人类已知的毒性最大的物质，可导致致命的神经麻痹性疾病肉毒中毒。肉毒杆菌中毒引起的弛缓性麻痹会持续几个月。肉毒毒素可作为潜在的生物恐怖制剂，对国土安全和公众健康构成巨大威胁。目前不存在用于肉毒中毒的预防性和治疗性治疗的有效且安全的解毒剂。唯一批准的解毒剂是马抗毒素，它只能中和血液循环中的毒素，并且治疗时间很短。此外，马抗毒素可引起严重的副作用，排除它们作为有效的促炎剂。病人唯一的选择仍然是在重症监护下进行人工通气几个月。在R21探索性研究基金中，我们提出了新一代肉毒杆菌中毒解毒剂，适体。适体是对其靶标具有高亲和力的独特寡核苷酸，其范围从小分子到蛋白质。适体具有低毒性、无报道的免疫原性、低制造成本和优异的储存稳定性。这些特性使适体成为肉毒杆菌中毒的理想解毒剂。本研究的目的是开发适体作为肉毒中毒的解毒剂。所提出的解毒剂有可能成为有效的预防性治疗和有效的治疗药物来治疗有症状的肉毒中毒患者。未来两年的具体目标是：（1）针对BoNT的适体的选择和表征。我们将使用SELEX来筛选随机RNA文库，分离针对BoNT的适体，并基于与BoNT的结合亲和力对适体进行优先排序。(2)适体的抗BoNT活性的验证。我们将使用体外和基于细胞的测定来确定对内肽酶活性和毒素与其受体结合的抑制作用。因此，从这项研究中鉴定的适体将被用作设计更有效的肉毒中毒解毒剂的先导化合物。这项研究将导致一种新的肉毒杆菌中毒解毒剂。该研究方法可以很容易地适用于所有七种血清型的BoNT和生物防御议程上的其他优先病原体。此外，这里提出的研究将对生物防御剂的快速诊断产生重大影响。 全世界生物恐怖主义造成的健康风险不断增加，给科学界带来了巨大负担，需要迅速开发生物和化学毒素的安全预防和治疗方法。这项任务一直是公共卫生的高度优先事项。肉毒杆菌中毒和大多数其他生物毒素（如NIAID优先病原体清单所列）缺乏治疗和预防药物。我们的项目将导致基于适体的肉毒中毒候选药物，肉毒中毒解毒剂的新类别，与目前可用的抗毒素有许多优势。