Inhibition of the β-Lactamase BlaMab by Avibactam Improves the In Vitro and In Vivo Efficacy of Imipenem against Mycobacterium abscessus
Inhibition of the β-Lactamase BlaMab by Avibactam Improves the In Vitro and In Vivo Efficacy of Imipenem against Mycobacterium abscessus
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DOI：
10.1128/aac.02440-16
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发表时间：
2017-04-01
影响因子：
4.9
通讯作者：
Mainardi, Jean-Luc
中科院分区：
文献类型：
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作者：
Lefebvre, Anne-Laure;Le Moigne, Vincent;Mainardi, Jean-Luc
分枝杆菌肺部感染用大环内酯类（克拉霉素或阿奇霉素）、氨基糖苷类（阿米卡星）和β-内酰胺类（头孢西丁或亚胺培南）治疗。三联组合在没有任何β-内酰胺酶抑制剂的情况下使用，即使M。该菌产生广谱β-内酰胺酶Bla（Mab）。我们确定阿维巴坦对BlaMab的抑制是否能提高亚胺培南对M.你好在肉汤和人巨噬细胞中测定药物组合的杀菌活性。通过监测感染的斑马鱼胚胎的存活来测试药物的体内功效。通过定量逆转录-PCR和Western印迹法比较肉汤和巨噬细胞中的BlaMab产生水平。亚胺培南（8或32 μ g/ml）、阿米卡星（32 μ g/ml）和阿维巴坦（4 μ g/ml）的三联组合在肉汤中具有杀菌作用（存活率< 0.1%），当亚胺培南以8和32 μ g/ml使用时，72 h时CFU数量分别减少3.2和4.3 log（10）。三重组合实现了显著的细胞内杀伤，低剂量（8 μ g/ml）和高剂量（32 μ g/ml）亚胺培南的细菌存活率分别为54%和7%。阿维巴坦对BlaMab的体内抑制改善了亚胺培南处理的斑马鱼胚胎的存活率。在感染的巨噬细胞中诱导编码BlaMab的基因的表达（20倍）。阿维巴坦对BlaMab的抑制可改善亚胺培南对M.在体外，巨噬细胞，和斑马鱼胚胎中，表明这种β-内酰胺酶抑制剂应进行临床评估。亚胺培南的体外评价可能低估了BlaMab的影响，因为β-内酰胺酶的产生在巨噬细胞中是可诱导的。
Mycobacterium abscessus pulmonary infections are treated with a macrolide (clarithromycin or azithromycin), an aminoglycoside (amikacin), and a beta-lactam (cefoxitin or imipenem). The triple combination is used without any beta-lactamase inhibitor, even though M. abscessus produces the broad-spectrum beta-lactamase Bla(Mab). We determine whether inhibition of BlaMab by avibactam improves the activity of imipenem against M. abscessus. The bactericidal activity of drug combinations was assayed in broth and in human macrophages. The in vivo efficacy of the drugs was tested by monitoring the survival of infected zebrafish embryos. The level of BlaMab production in broth and in macrophages was compared by quantitative reverse transcription-PCR and Western blotting. The triple combination of imipenem (8 or 32 mu g/ml), amikacin (32 mu g/ml), and avibactam (4 mu g/ml) was bactericidal in broth (< 0.1% survival), with 3.2- and 4.3-log(10) reductions in the number of CFU being achieved at 72 h when imipenem was used at 8 and 32 mu g/ml, respectively. The triple combination achieved significant intracellular killing, with the bacterial survival rates being 54% and 7% with the low (8 mu g/ml) and high (32 mu g/ml) dosages of imipenem, respectively. In vivo inhibition of BlaMab by avibactam improved the survival of zebrafish embryos treated with imipenem. Expression of the gene encoding BlaMab was induced (20-fold) in the infected macrophages. Inhibition of BlaMab by avibactam improved the efficacy of imipenem against M. abscessus in vitro, in macrophages, and in zebrafish embryos, indicating that this beta-lactamase inhibitor should be clinically evaluated. The in vitro evaluation of imipenem may underestimate the impact of BlaMab, since the production of the beta-lactamase is inducible in macrophages.