Nuclease-Resistant Aptamers to Botox and Anthrax Toxins

肉毒杆菌和炭疽毒素的核酸酶抗性适体

基本信息

批准号：
6736672
负责人：
John G Bruno
金额：
$ 9.99万
依托单位：
OPERATIONAL TECHNOLOGIES CORPORATION
依托单位国家：
美国
项目类别：
财政年份：
2004
资助国家：
美国
起止时间：
2004-04-15 至 2004-10-14
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/6736672
关键词：
anthrax toxin binding sites biotechnology bioterrorism /chemical warfare botulinum toxins drug design /synthesis /production endonuclease exonuclease genetic library nucleic acid inhibitor nucleic acid structure oligonucleotides polymerase chain reaction protein binding

项目摘要

DESCRIPTION (provided by investigator): Botulinum toxin and anthrax are two likely bioweapons of choice in military attacks or terrorism against the United States. As such, effective post-exposure therapies, such as antitoxins, to both agents are required. Operational Technologies (OpTech) has extensive experience in the area of DNA aptamer development for the detection of biowarfare agents and biotoxins via its Senior Scientist. OpTech proposes to team with Prof. James Chambers of the Biochemistry Department at the University of Texas at San Antonio (UTSA). Together, the OpTech-UTSA team will develop 2'-pyrimidine modified nuclease-resistant "shielded" DNA aptamers by the SELEX process to botulinum toxin (BOTOX) and the three anthrax toxin components (edema factor, lethal factor, and protective antigen). "Shielded aptamers" are well suited to eventual in vivo therapeutic use and would be of low immunogenicity. In Phase I, OpTech will generate shielded aptamers to several serotypes (at least A, B and E) of BOTOX and will determine their relative binding affinities for the intact "derivative" toxin as well as the separate heavy (100 kD) and light 50 kD) chains. In Phase II, OpTech will develop and test anti-anthrax toxin component shielded aptamers as well as clone and sequence all promising aptamers reactive against either toxin family. Development of "shielded" (nuclease-resistant) aptamers as toxin inhibitors with greater affinity than antibodies or other antidotes, in general, guarantees the owner of the technology entry into a growing biodefense market. If successful in this project, OpTech will use the knowledge gained to develop improved inhibitors of other bacterial toxins, such as cholera toxin, as well as snake and spider venoms. The therapeutic market for such products would be immense, because DNA is nontoxic and hypoallergenic (does not induce a significant immune response), thus avoiding "serum sickness" or anaphylaxis upon secondary use. Such reagents could also be used in environmental biosensors or diagnostic test kits for detection of BOTOX and anthrax toxins in body fluids or environmental samples.

描述（调查员提供）：肉毒杆菌毒素和炭疽病是军事袭击或针对美国恐怖主义的两个可能选择的生物武器。因此，需要对两种药物等有效的暴露后疗法，例如抗毒素。操作技术（OPTECH）在DNA适体开发领域具有丰富的经验，可通过其高级科学家检测生物处理剂和生物毒素。 Optech建议与得克萨斯大学圣安东尼奥分校（UTSA）的生物化学系的詹姆斯·钱伯斯教授团队合作。 Optech-UTSA团队将通过SELEX过程开发2'-吡啶定修饰的核酸酶耐药性“屏蔽” DNA适体，到肉毒杆菌毒素（肉毒杆菌毒素）和三种炭疽毒素成分（表育素性因子，致死因子和保护性抗原）。 “屏蔽的适体”非常适合最终的体内治疗用途，并且免疫原性低。在第一阶段，Optech将对肉毒杆菌毒素的几种血清型（至少A，B和E）产生屏蔽的适体，并将确定其对完整“衍生物”毒素以及独立的重量（100 kd）和50 kD的相对结合亲和力。在第二阶段，Optech将开发和测试抗蒽毒素成分屏蔽适体以及克隆和序列和序列所有有望对任何一种毒素家族反应反应的适体。通常，与抗体或其他解毒剂相比，“屏蔽”（耐核酸酶的）适体的发展，通常可以保证技术进入增长的生物化市场。如果在该项目中取得成功，Optech将利用获得的知识来发展其他细菌毒素的抑制剂，例如霍乱毒素，以及蛇和蜘蛛毒液。此类产品的治疗市场将是巨大的，因为DNA是无毒的和低过敏性的（不会诱导明显的免疫反应），因此避免了二次使用后“血清疾病”或过敏反应。此类试剂也可以用于环境生物传感器或诊断测试试剂盒中，以检测体液或环境样品中的肉毒杆菌毒素和炭疽毒素。