Gram-negative bacterial infections are a significant public health concern, and the lack of new drug classes for these pathogens is linked to the inability of most drug leads to accumulate inside Gram-negative bacteria1-7. Here, we report the development of a web application-eNTRyway-that predicts compound accumulation (in Escherichia coli) from its structure. In conjunction with structure-activity relationships and X-ray data, eNTRyway was utilized to re-design Debio-1452-a Gram-positive-only antibiotic8-into versions that accumulate in E. coli and possess antibacterial activity against high-priority Gram-negative pathogens. The lead compound Debio-1452-NH3 operates as an antibiotic via the same mechanism as Debio-1452, namely potent inhibition of the enoyl-acyl carrier protein reductase FabI, as validated by in vitro enzyme assays and the generation of bacterial isolates with spontaneous target mutations. Debio-1452-NH3 is well tolerated in vivo, reduces bacterial burden in mice and rescues mice from lethal infections with clinical isolates of Acinetobacter baumannii, Klebsiella pneumoniae and E. coli. This work provides tools for the facile discovery and development of high-accumulating compounds in E. coli, and a general blueprint for the conversion of Gram-positive-only compounds into broad-spectrum antibiotics.

中文翻译：

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渗透规则的实施导致 FabI 抑制剂具有抗革兰氏阴性病原体的活性。

革兰氏阴性细菌感染是一个重要的公共卫生问题，缺乏针对这些病原体的新药物类别与大多数药物无法在革兰氏阴性细菌内部积聚有关1-7。在这里，我们报告了一个 Web 应用程序 eNTRyway 的开发，该应用程序从其结构预测化合物积累（在大肠杆菌中）。结合构效关系和 X 射线数据，eNTRyway 被用来将 Debio-1452-一种仅革兰氏阳性抗生素8-重新设计成在大肠杆菌中积累并具有针对高优先级革兰氏阳性菌的抗菌活性的版本。阴性病原体。先导化合物 Debio-1452-NH3 作为抗生素通过与 Debio-1452 相同的机制发挥作用，即有效抑制烯酰基-酰基载体蛋白还原酶 FabI，通过体外酶分析和具有自发靶点突变的细菌分离株的产生进行验证。Debio-1452-NH3 在体内具有良好的耐受性，可减少小鼠的细菌负担，并使小鼠免受鲍曼不动杆菌、肺炎克雷伯菌和大肠杆菌临床分离株的致命感染。这项工作为在大肠杆菌中轻松发现和开发高积累化合物提供了工具，并为将仅革兰氏阳性化合物转化为广谱抗生素提供了总体蓝图。