In Vitro Core

该中心将应用最先进的基因组学和代谢组学方法，指导发现生物多样性资源开发不足产生的高价值线索。我们将专注于通过微生物和宿主之间的共生通信的调解而进化为功能性的化合物。体外核心（核心B）科学家将与Bugni教授的实验室合作，生产和纯化中心所有三个项目提供的天然产品。核心B科学家将通过利用目前在UW小分子筛选和合成设施（SMSSF）和Bugni教授实验室提供的高通量检测基础设施和专业知识，评估提取物，馏分和化合物对耐药目标病原体的抗菌活性。将评价抗微生物测定中的剂量反应特性，以优先选择具有广泛杀灭活性的最有效化合物，该化合物可导致培养物中微生物细胞死亡。然后将评估具有抗微生物活性的化合物对原代人类细胞的细胞毒性，以便在项目早期消除有毒化合物。核心B科学家还将负责使用Bugni教授实验室开发的优化发酵方法和生产规模纯化基础设施扩大纯化合物的生产（由中心PI和项目负责人选择）。化合物将通过与华盛顿大学药学院药物实验站的协调来评估其稳定性、溶解度和配方。将向核心C和核心D（作用机制核心）提供足量的纯化合物，以测试化合物在小鼠中的疗效和安全性。核心B将提供药物化学专业知识，以评估简化类似物和/或半合成衍生化的潜在合成策略，以改善支架的药代动力学或确定构效关系。

The Center will apply state-of-the-art genomics and metabolomics approaches to guide the discovery of high value leads produced by under-explored sources of biological diversity. We will focus on compounds that have evolved to be functional through mediation of symbiotic communication between microbes and hosts. The In vitro Core (Core B) scientists will work collaboratively with Professor Bugni's laboratory to produce and purify natural products provided by all three projects in the Center. Core B scientists will assess the anti-microbial activities of extracts, fractions and compounds against drug resistant target pathogens by leveraging the high throughput assay infrastructure and expertise currently available in the UW Small Molecule Screening and Synthesis Facility (SMSSF) and Professor Bugni's laboratory. Doseresponse properties in the antimicrobial assays will be evaluated to prioritize the selection ofthe most potent compounds with broad, cidal activity that causes the death of microbial cells in culture. Compounds with antimicrobial activity then will be assessed for cytotoxicity on primary human cells in order to eliminate toxic compounds early in the projects. Core B scientists also will be responsible for scale-up production of pure compounds (selected by the Center PI and Project leaders) using optimized fermentation approaches developed in Professor Bugni's laboratory and production-scale purification infrastructure. Compounds will be evaluated for stability, solubility and formulation through coordination with the UW School of Pharmacy Pharmaceutical Experiment Station. Sufficient quantities ofthe pure compounds will be provided to Core C for testing the efficacy and safety of the compounds in mice and to Core D, the Mechanism of Action Core. Core B will provide medicinal chemistry expertise to evaluate potential synthetic strategies for simplified analogs and/or for semisynthetic derivatization to improve the pharmacokinetics of scaffolds or determine structure-activity-relationships.