Pathogen-specific drug targets for weaponized bacteria

描述(由申请人提供)：对当前抗生素治疗具有抗药性的工程细菌生物武器的威胁为开发新型治疗化合物提供了强大的授权。这项提案是洛克菲勒大学三个互补小组(文森特·费舍蒂博士、亚历山大·托马兹博士和埃里克·斯特宾斯博士)共同努力，以确定A类病原体、炭疽杆菌和鼠疫杆菌以及B类食品和水安全威胁的新型抑制剂。第一个也是中心的研究项目是基于这样的观察，即对噬菌体裂解酶(例如炭疽杆菌的PlyG)的抗药性是极其罕见的事件，由此我们假设这些酶的结合区域针对细菌细胞壁中的一个关键成分以确保噬菌体的释放(从某种意义上说，噬菌体通过进化进行了高通量分析来鉴定这个成分)。因此，合成这种壁组分的途径代表了抗生素开发的目标。因此，核心项目将集中于裂解酶结合底物的鉴定、分离和表征。一旦确定，其合成的途径将被确定。这些催化该途径的酶将被分离、纯化和结晶，并确定抑制物。Alex Tomasz的细胞壁结构经验为核心项目提供了素材，但也将其扩大到通过对细胞壁的高分辨率分析和通过分离条件致命突变体-使用水手类型元素的体内转座-来寻找其他针对炭疽杆菌的新靶点。细胞壁在他的项目中占据了中心舞台，因为(I)PlyG的目标是在壁中；(Ii)细胞壁在炭疽杆菌感染引起的毁灭性败血症休克中发挥重要作用，以及(Iii)炭疽杆菌基因组包含与壁相关的决定因素，这些决定因素在其他细菌中是毒力因子。EREC Stebbins将提供蛋白质结构和结晶方面的专业知识，以及识别这些结构的小分子抑制剂。他的项目通过在计算药物发现与结构确定相结合的计算中的翻译成分，将新的疗法搜索扩展到包括鼠疫杆菌和B类病原体。总体而言，这些努力针对的是关键生物武器病原体的中心毒力因子。

DESCRIPTION (provided by applicant): The threat of engineered bacterial bioweapons that are resistant to current antibiotic treatments provides a powerful mandate to develop novel classes of therapeutic compounds. This proposal presents a concerted effort of three complementary groups at the Rockefeller University (Dr. Vincent Fischetti, Dr. Alexander Tomasz, and Dr. Erec Stebbins) to identify novel inhibitors of the Category A pathogens, B. anthracis and Y. pestis, as well as Category B food and water safety threats. The first and central research project is based on the observation that resistance to bacteriophage lytic enzymes, such as PlyG for B. anthracis, is an extremely rare event, from which we hypothesize that the binding domain of these enzymes targets a critical component in the bacterial cell wall to assure phage release (in a sense through evolution, the phage have performed the high throughput assay to identify this component). Thus, the pathway to the synthesis of this wall component represents a target for antibiotic development. The core project will therefore focus on the identification, isolation and characterization of the lytic enzyme binding substrate. Once identified, the pathway towards its synthesis will be determined. Those enzymes catalyzing the pathway will be isolated, purified and crystallized, and inhibitors identified. The cell wall structural experience of Alex Tomasz, feeds into the core project but also broadens it to a search for additional novel targets against B. anthracis by high resolution analysis of the cell walls and by the isolation of conditional lethal mutants - using in vivo transposition of a mariner type element. Cell walls occupy center stage in his project since (i) the target of PlyG is in the wall; (ii) cell walls play an important role in the devastating septic shock caused by B. anthracis infections and (iii) the genome of B. anthracis contains wall-related determinants that are virulence factors in other bacteria. Erec Stebbins will provide the expertise in protein structure and crystallization along with the identification of small molecule inhibitors for these structures. His project expands the novel therapeutics search to include Y. pestis and Category B pathogens through a translational component in computation drug discovery coupled to structural determination. In total, these efforts target central virulence factors in critical biowarfare pathogens.