Isoprenoids are vital for all organisms, in which they maintain membrane stability and support core functions such as respiration1. IspH, an enzyme in the methyl erythritol phosphate pathway of isoprenoid synthesis, is essential for Gram-negative bacteria, mycobacteria and apicomplexans2,3. Its substrate, (E)-4-hydroxy-3-methyl-but-2-enyl pyrophosphate (HMBPP), is not produced in metazoans, and in humans and other primates it activates cytotoxic Vγ9Vδ2 T cells at extremely low concentrations4-6. Here we describe a class of IspH inhibitors and refine their potency to nanomolar levels through structure-guided analogue design. After modification of these compounds into prodrugs for delivery into bacteria, we show that they kill clinical isolates of several multidrug-resistant bacteria-including those from the genera Acinetobacter, Pseudomonas, Klebsiella, Enterobacter, Vibrio, Shigella, Salmonella, Yersinia, Mycobacterium and Bacillus-yet are relatively non-toxic to mammalian cells. Proteomic analysis reveals that bacteria treated with these prodrugs resemble those after conditional IspH knockdown. Notably, these prodrugs also induce the expansion and activation of human Vγ9Vδ2 T cells in a humanized mouse model of bacterial infection. The prodrugs we describe here synergize the direct killing of bacteria with a simultaneous rapid immune response by cytotoxic γδ T cells, which may limit the increase of antibiotic-resistant bacterial populations.

中文翻译：

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IspH 抑制剂杀死革兰氏阴性菌并调动免疫清除

类异戊二烯对所有生物体都至关重要，它们在其中维持膜稳定性并支持呼吸等核心功能1。IspH 是类异戊二烯合成的甲基赤藓糖醇磷酸途径中的一种酶，对革兰氏阴性菌、分枝杆菌和顶端复合菌至关重要 2,3。它的底物 (E)-4-hydroxy-3-methyl-but-2-enyl pyrophosphate (HMBPP) 在后生动物中不产生，在人类和其他灵长类动物中，它在极低浓度下激活细胞毒性 Vγ9Vδ2 T 细胞4-6。在这里，我们描述了一类 IspH 抑制剂，并通过结构引导的类似物设计将其效力提高到纳摩尔水平。在将这些化合物修饰为前体药物以输送到细菌中后，我们发现它们可以杀死几种耐多药细菌的临床分离物，包括不动杆菌属、假单胞菌属、克雷伯菌属、肠杆菌属、弧菌属、志贺氏菌属、沙门氏菌属、耶尔森氏菌属、分枝杆菌属和芽孢杆菌属——但对哺乳动物细胞相对无毒。蛋白质组学分析表明，用这些前药处理的细菌与条件性 IspH 敲低后的细菌相似。值得注意的是，这些前药还在细菌感染的人源化小鼠模型中诱导人 Vγ9Vδ2 T 细胞的扩增和活化。我们在此描述的前药协同直接杀死细菌和细胞毒性 γδ T 细胞的同时快速免疫反应，这可能会限制抗生素抗性细菌种群的增加。蛋白质组学分析表明，用这些前药处理的细菌与条件性 IspH 敲低后的细菌相似。值得注意的是，这些前药还在细菌感染的人源化小鼠模型中诱导人 Vγ9Vδ2 T 细胞的扩增和活化。我们在此描述的前药协同直接杀死细菌和细胞毒性 γδ T 细胞的同时快速免疫反应，这可能会限制抗生素抗性细菌种群的增加。蛋白质组学分析表明，用这些前药处理的细菌与条件性 IspH 敲低后的细菌相似。值得注意的是，这些前药还在细菌感染的人源化小鼠模型中诱导人 Vγ9Vδ2 T 细胞的扩增和活化。我们在此描述的前药协同直接杀死细菌和细胞毒性 γδ T 细胞的同时快速免疫反应，这可能会限制抗生素抗性细菌种群的增加。