Identification of macrocyclic molecules with antibacterial activity

项目摘要 我们开发了一种新的方法来鉴定具有抗菌活性的分子 快速区分不同的作用机制(MOA)。这种方法， 细菌细胞学分析(BCP)，使用定量荧光显微镜 测量抗生素治疗对单个细胞的影响。以抗生素为目标的 不同的细胞途径和途径中的不同步骤产生独特的 细胞学特征，允许识别可能的新化合物的MOA 几个小时。Linneeus Bioscience Inc.是一家初创公司，成立目的是将 这项技术并使其可在收费的基础上向制药商提供服务 工业界和科学界。在这个第一阶段的项目中，Linneeus将筛选 最初开发的包含6,000多种大环化合物的高度多样化的库 作为激酶抑制剂，以识别靶向多药耐药的新分子 (MDR)革兰氏阴性菌。这一系列独特的化学支架从未 已经对抗菌化合物进行了筛选，初步数据显示它是 富含特定MOA的抗菌分子。我们将对这些藏品进行筛选 抗野生型大肠杆菌。我们将使用BCP来确定所有点击的可能的MOA 在确定优先顺序之前，并快速消除那些不特定或不受欢迎的 活性，如对膜的洗涤剂样作用。我们将进行反筛选 针对真核细胞系的细胞毒性，消除剧毒分子。我们会 使用经典的分子遗传学和生化方法来鉴定准确的 优先命中的分子靶点，并评估对 浮出水面。我们将确定最易处理的化学优先热门系列并生成 早期通过药物化学努力取得领先地位。这些研究的目标是确定 可以进一步发展的早期铅分子。这些研究将证明 BCP是一种基于全细胞成像的健壮方法，能够通过 工业规模的抗菌性能。这个平台将被广泛地制作 以服务费形式提供。

Project Summary We have developed a new method to identify molecules with antibacterial activities that rapidly discriminates between different mechanisms of action (MOA). This approach, bacterial cytological profiling (BCP), uses quantitative fluorescence microscopy to measure the effects of antibiotic treatment on individual cells. Antibiotics that target different cellular pathways and different steps within a pathway generate unique cytological profiles, allowing identification of the likely MOA of new compounds within a few hours. Linnaeus Bioscience Inc. is a start up company founded to commercialize this technology and make it accessible on a fee for service basis to the pharmaceutical industry and the scientific community. During this Phase 1 project, Linnaeus will screen a highly diverse library of more than 6,000 macrocyclic compounds originally developed as kinase inhibitors in order to identify new molecules targeting multidrug resistant (MDR) Gram negative bacteria. This collection of unique chemical scaffolds has never been screened for antibacterial compounds and our preliminary data shows that it is enriched for antibacterial molecules with specific MOAs. We will screen this collection against wild type E. coli. We will employ BCP to identify the likely MOA for all hits before prioritization, and to quickly eliminate those with nonspecific or undesired activities, such as detergent-like effects on the membrane. We will counter screen against eukaryotic cell lines for cytotoxicity to eliminate highly toxic molecules. We will use classical molecular genetic and biochemical approaches to identify the precise molecular target of the prioritized hits, and to evaluate the potential for resistance to emerge. We will identify the most chemically tractable priority hit series and generate early leads through medicinal chemistry efforts. The goal of these studies is to identify early lead molecules that can be developed further. These studies will demonstrate that BCP is a robust whole cell imaging based method capable of identifying molecules with antibacterial properties on an industrial scale. This platform will be made widely available on a fee for service basis.