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耐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有力的要求。该提案提出了洛克菲勒大学三个互补小组（Vincent Fischetti博士、Alexander Tomasz博士和Erec Stebbins博士）的共同努力，以确定a类病原体（B类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。第一个中心研究项目是基于对噬菌体裂解酶（如炭疽杆菌的PlyG）的耐药性的观察，这是一种极其罕见的事件，由此我们假设这些酶的结合域靶向细菌细胞壁上的一个关键成分，以确保噬菌体释放（从某种意义上说，通过进化，噬菌体已经执行了高通量测定来识别该成分）。因此，这种壁成分的合成途径代表了抗生素开发的目标。因此，核心项目将侧重于裂解酶结合底物的鉴定、分离和表征。一旦确定，其合成途径将被确定。催化该途径的酶将被分离、纯化和结晶，并确定抑制剂。Alex Tomasz的细胞壁结构经验不仅为核心项目提供了信息，而且还通过对细胞壁的高分辨率分析和使用mariner型元素的体内转位分离条件致死突变体，将其扩展到寻找针对炭疽芽孢杆菌的其他新靶标。细胞壁在他的项目中占据中心位置，因为(i) PlyG的靶点在细胞壁中；（ii）细胞壁在炭疽芽孢杆菌感染引起的毁灭性感染性休克中起重要作用；（iii）炭疽芽孢杆菌的基因组包含与细胞壁相关的决定因素，这些决定因素在其他细菌中是毒力因子。eric Stebbins将提供蛋白质结构和结晶方面的专业知识，以及这些结构的小分子抑制剂的鉴定。他的项目扩展了新疗法的搜索，包括鼠疫杆菌和B类病原体，通过计算药物发现和结构确定的翻译组成部分。总的来说，这些努力的目标是关键生物战病原体的核心毒力因子。

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.