LN-1-255, a penicillanic acid sulfone able to inhibit the class D carbapenemase OXA-48.
LN-1-255, a penicillanic acid sulfone able to inhibit the class D carbapenemase OXA-48.
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LN-1-255，一种青霉烷酸砜，能够抑制 D 类碳青霉烯酶 OXA-48。
DOI：
10.1093/jac/dkw105
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发表时间：
2016
期刊：
影响因子：
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通讯作者：
Beceiro,Alejandro
中科院分区：
文献类型：
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作者：
Vallejo,JuanA;Martínez-Guitián,Marta;Vázquez-Ucha,JuanC;González-Bello,Concepción;Poza,Margarita;Buynak,JohnD;Bethel,ChristopherR;Bonomo,RobertA;Bou,German;Beceiro,Alejandro
目的碳青霉烯酶是肠杆菌科细菌对碳青霉烯类抗生素耐药的重要机制。在碳青霉烯酶中，OXA-48具有独特的挑战，因为它对β-内酰胺类抑制剂具有耐药性。在此，我们测试了化合物LN-1-255对碳青霉烯酶OXA-48活性的抑制能力。LN-1-255是一种6-亚烷基-2‘-取代青霉酸砜。方法克隆OXA-48基因并在肺炎克雷伯菌和大肠埃希菌中表达，用克拉维酸、他唑巴坦、舒巴坦和LN-1-255诱导MICs。纯化了OXA-48，用LN-1-255和他唑巴坦进行了稳态动力学研究。通过对接实验研究了LN-1-255与OXA-48的共价结合方式。结果4 mg/L LN-1-255和16 mg/L LN-1-255分别使OXA-48临床株和转化株对碳青霉烯类抗生素的敏感性增加2~32倍和4倍。动力学实验表明，LN-1-255能抑制OxA-48，其酰化效率(K_2/K)为10±1×10~4M−_1·S−_1，慢去酰化速率(K_(OFF))为7±1×10~(-1)−_4 S−_1。最后，LN-1-255的kcat/kinact比他唑巴坦低500倍。结论在本研究中，碳青霉烯类抗生素与LN-1-255联合使用对碳青霉烯酶OXA-48有效，OXA-48是一种重要的抗生素耐药机制。这为进一步研究青霉素对携带D类质粒的肠杆菌科碳青霉烯酶的抑制作用提供了动力。
Objectives Carbapenemases are the most important mechanism responsible for carbapenem resistance in Enterobacteriaceae. Among carbapenemases, OXA-48 presents unique challenges as it is resistant to β-lactam inhibitors. Here, we test the capacity of the compound LN-1-255, a 6-alkylidene-2′-substituted penicillanic acid sulfone, to inhibit the activity of the carbapenemase OXA-48. Methods The OXA-48 gene was cloned and expressed in Klebsiella pneumoniae and Escherichia coli in order to obtain MICs in the presence of inhibitors (clavulanic acid, tazobactam and sulbactam) and LN-1-255. OXA-48 was purified and steady-state kinetics was performed with LN-1-255 and tazobactam. The covalent binding mode of LN-1-255 with OXA-48 was studied by docking assays. Results Both OXA-48-producing clinical and transformant strains displayed increased susceptibility to carbapenem antibiotics in the presence of 4 mg/L LN-1-255 (2–32-fold increased susceptibility) and 16 mg/L LN-1-255 (4–64-fold increased susceptibility). Kinetic assays demonstrated that LN-1-255 is able to inhibit OXA-48 with an acylation efficiency (k2/K) of 10±1× 104 M− 1 s− 1 and a slow deacylation rate (koff) of 7±1× 10− 4 s− 1. IC50 was 3 nM for LN-1-255 and 1.5 μM for tazobactam. Lastly, kcat/kinact was 500-fold lower for LN-1-255 than for tazobactam. Conclusions In these studies, carbapenem antibiotics used in combination with LN-1-255 are effective against the carbapenemase OXA-48, an important emerging mechanism of antibiotic resistance. This provides an incentive for further investigations to maximize the efficacy of penicillin sulfone inhibition of class D plasmid-carried Enterobacteriaceae carbapenemases.