Small Molecule Inhibitors of C. perfringens Epsilon-Toxin

描述（由申请人提供）：SBIR第一阶段提案的总体目标是找到抑制C的化合物。使用Innovative Biologics，Inc.开发的用于灭活成孔毒素的新方法，它基于用具有与孔本身相同对称性的分子阻塞目标孔。以前，我们证明了具有七重对称性的β-环糊精（β-CD）的全取代衍生物可以阻断由炭疽杆菌保护性抗原（PA）形成的七聚体孔，并且在基于细胞的测定和动物试验中对炭疽致死毒素作用具有保护作用。自从C.由于产气荚膜杆菌β-毒素形成类似于PA孔的七聚体跨膜孔，我们提出筛选我们的全取代β-CD衍生物文库以获得β-毒素活性的抑制剂。本I期研究的具体目标是：（1）建立和验证用于测试β-CD衍生物阻断C形成的孔的能力的测定方法。产气荚膜杆菌毒素并抑制其细胞毒活性。(2)测试来自全取代β-CD衍生物代表性库的化合物的阻断和抑制活性，并选择最有效的化合物作为抗毒素药物进行进一步开发。如果在这项可行性研究中发现了有效的C-毒素抑制剂，在第二阶段，我们将利用我们的初步测试数据和C-毒素的晶体结构信息。本发明涉及与计算化学协同使用产气荚膜杆菌β-毒素来设计对β-毒素孔具有增强的亲和力的另外的β-CD衍生物。设计的化合物将在体内和体外合成和测试，最终目标是找到新的治疗C。产气荚膜杆菌毒素。项目叙述：产气荚膜梭菌产生的埃帕霉素毒素是最致命的细菌毒素之一。它被视为一种潜在的生物武器，被列入B类优先物剂清单。目前，没有有效的治疗方法用于抗毒素介导的中毒;因此，非常需要开发针对这种生物防御毒素的治疗剂。

DESCRIPTION (provided by applicant): The overall goal of this SBIR Phase I proposal is to find compounds that inhibit C. perfringens epsilon-toxin action using a novel approach for the inactivation of pore-forming toxins developed at Innovative Biologics, Inc. It is based on the blocking of the target pore with molecules having the same symmetry as the pore itself. Previously, we demonstrated that per-substituted derivatives of beta-cyclodextrin (beta-CD) having a sevenfold symmetry could block the heptameric pore formed by Bacillus anthracis protective antigen (PA) and were protective against anthrax lethal toxin action in cell-based assays and in animals tests. Since C. perfringens epsilon -toxin forms a heptameric trans-membrane pore similar to the PA pore, we propose to screen our library of per-substituted beta-CD derivatives for inhibitors of epsilon - toxin's activity. The specific aims of this Phase I study are: (1) Establish and validate assays for testing the ability of beta -CD derivatives to block the pore formed by C. perfringens epsilon -toxin and to inhibit its cytotoxic activity. (2) Test blocking and inhibitory activity of compounds from a representative library of per-substituted beta -CD derivatives and select the most potent compounds for further development as anti-toxin drugs. Provided that effective inhibitors of epsilon -toxin are found in this feasibility study, in Phase II we will utilize our initial testing data and the crystal structure information available for C. perfringens epsilon -toxin in concert with computational chemistry to design additional beta-CD derivatives with enhanced affinity to the epsilon -toxin pore. The designed compounds will be synthesized and tested in vivo and in vitro with the eventual goal of finding new therapeutics against C. perfringens epsilon -toxin. Project Narrative: Epsilon toxin produced by Clostridium perfringens is one of the most lethal bacterial toxins. It is regarded as a potential biological weapon and is included in the list of category B priority agents. Currently, there is no effective treatment for the epsilon -toxin-mediated intoxication; therefore, a great need exists for the development of therapeutics against this biodefense toxin.