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生物恐怖主义的6种最高风险威胁因子之一(A类制剂)，因为它们具有极强的效力和致命性，易于生产和运输，需要长期的重症监护。流氓国家制造和武器化了核弹，恐怖组织部署了核弹。由于这些威胁，需要特定的药剂来预防和治疗中毒。唯一在动物和人类中得到验证的治疗肉毒杆菌中毒的近期产品是抗毒素。在之前的NIAID资助下，我们已经为A型、B型和E型BONT产生了一种有效的人类相容的重组单抗抗毒素，目前正在单独的NIAID资助下开发用于人体临床试验的BONT/A重组抗毒素。这项建议的总体目标是产生可用于预防和治疗由BONT血清型C、D、F和G型引起的肉毒杆菌中毒的新型人类相容单抗(MAbs)。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)生成针对这四种BONT血清型的广泛中和的人类相容mAb；以及3)关于血清型交叉反应性和毒素中和作用的mAb的体外和体内表征。这些目标将由在该领域工作超过13年的肉毒杆菌中毒专家组成的跨机构团队实现：詹姆斯·D·马克斯、抗体工程和伦纳德·史密斯、BONT疫苗开发、毒素和毒素片段的产生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微生物遗传特征专家凯伦·希尔的协助，他将确定毒素多样性的程度，以确保抗体中和所有可能的毒素亚型。在资助期结束时，我们预计已经产生了针对C、D、F和G型BONT的重组抗毒素，可以转移到制造和后期开发过程中。结合以前的结果，这将使安全、有效的人类相容抗毒素适用于所有七种BONT血清型。

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.