Development of botulinum neurotoxin immunotherapy, serotypes C, D, F, and G

描述（由申请人提供）：肉毒杆菌神经毒素（BoNTs）被疾病控制中心列为6种最高风险生物恐怖主义威胁制剂（“A类制剂”）之一，因为它们具有极强的效力和致命性，易于生产和运输，并且需要长期的重症监护。流氓国家生产和武器化了导弹，恐怖组织部署了导弹。由于这些威胁，需要特定的药物来预防和治疗中毒。治疗肉毒杆菌中毒的唯一近期产品已在动物和人类中得到验证是抗毒素。在NIAID之前的资助下，我们已经为BoNT血清型a、B和e产生了一种有效的人类兼容重组单克隆抗体抗毒素。BoNT/ a重组抗毒素目前正在NIAID的单独资助下进行人体临床试验。本提案的总体目标是产生新的人类兼容单克隆抗体（mab），可用于预防和治疗由BoNT血清型C、D、F和G引起的肉毒中毒。实现这一目标需要三个组成部分：1)确定这4种毒素血清型（C、D、F和G）的BoNT基因多样性程度；2)对四种BoNT血清型产生广泛中和的人兼容单克隆抗体；3)单抗血清型交叉反应性和毒素中和的体内外鉴定。这些目标将由在该领域工作超过13年的肉毒杆菌专家组成的机构间团队实现：James D. Marks，抗体工程和Leonard Smith， BoNT疫苗开发，毒素和毒素片段生成以及毒素中和的动物模型。他们将得到洛斯阿拉莫斯国家实验室（Los Alamos National Laboratory）微生物遗传特征专家凯伦·希尔（Karen Hill）的协助，她将确定毒素多样性的程度，以确保抗体能中和所有可能的毒素亚型。在融资期结束时，我们预计将产生针对BoNT血清型C、D、F和G的重组抗毒素，该抗毒素可以转移到生产和后期开发途径。结合以往的结果，这将为所有7种BoNT血清型提供一种安全、有效的人类兼容抗毒素。

DESCRIPTION (provided by applicant): Botulinum neurotoxins (BoNTs) are classified by the Centers for Disease Control as one of the 6 highest-risk threat agents for bioterrorism (the "Category A agents"), due to their extreme potency and lethality, ease of production and transport, and need for prolonged intensive care. BoNTs have been produced and weaponized by rogue nations and deployed by terrorist groups. As a result of these threats, specific pharmaceutical agents are needed for prevention and treatment of intoxication. The only near term product for treatment of botulism that has been validated in animals and humans is antitoxin. Under previous NIAID funding, we have generated a potent human compatible recombinant monoclonal antibody based antitoxin for BoNT serotypes A, B, and E. The BoNT/A recombinant antitoxin is currently being developed under separate NIAID funding for human clinical trials. The overall goal of this proposal is to generate novel human compatible monoclonal antibodies (mAbs) that can be used to prevent and treat botulism caused by BoNT serotypes C, D, F, and G. Achievement of this goal requires three components: 1) determination of the extent of BoNT gene diversity of these 4 toxin serotypes (C, D, F, and G); 2) generation of panels of broadly neutralizing human compatible mAbs to the four BoNT serotypes; and 3) in vitro and in vivo characterization of mAbs with respect to serotype cross reactivity and toxin neutralization. These goals will be achieved by an inter-institutional team of botulism experts who have worked in this field for more than 13 years: James D. Marks, antibody engineering and Leonard Smith, BoNT vaccine development, toxin and toxin fragment generation and animal models of toxin neutralization. They will be assisted by an expert in microbial genetic characterization, Karen Hill, at Los Alamos National Laboratory, who will define the extent of toxin diversity to ensure that antibodies neutralize all possible toxin subtypes. At the end of the funding period we anticipate having generated a recombinant antitoxin for BoNT serotypes C, D, F, and G that can be transferred into the manufacturing and late stage development pathway. Combined with previous results, this will make available a safe, efficacious human compatible antitoxin for all seven BoNT serotypes.