Validation of a High-Throughput Screening Assay for Identification of Adjunctive and Directly Acting Antimicrobials Targeting Carbapenem-Resistant Enterobacteriaceae.
Validation of a High-Throughput Screening Assay for Identification of Adjunctive and Directly Acting Antimicrobials Targeting Carbapenem-Resistant Enterobacteriaceae.
复制标题
验证针对碳青霉烯类耐药肠杆菌的辅助和直接作用抗菌药物的高通量筛选测定。
DOI：
10.1089/adt.2016.701
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发表时间：
2016
影响因子：
1.8
通讯作者：
Kirby,JamesE
中科院分区：
文献类型：
--
作者：
Smith,KennethP;Kirby,JamesE
We describe development and validation of ahigh-throughput screen (HTS) for identifying small molecules that restore the efficacy of carbapenems (adjunctives) and/or directly inhibit growth of carbapenem-resistant Enterobacteriaceae (CRE). Our HTS assay is based on a screen–counterscreen approach using a representative multidrug-resistant CRE strain,Klebsiella pneumoniaeBIDMC12A. Specifically, we tested the ability of small molecules to inhibit bacterial growth in the presence (screen) or absence (counterscreen) of meropenem, a representative carbapenem antibiotic. Primary screening of 11,698 known bioactive compounds identified 14 with adjunctive activity and 79 with direct antimicrobial effect. Secondary screening identified triclosan as a strongly synergistic meropenem adjunctive (fractional inhibitory concentration = 0.48) and confirmed azidothymidine (AZT) (minimal inhibitory concentration [MIC] = 4 μg mL−1), NH125 (MIC = 4 μg mL−1), diphenyleneiodonium chloride (MIC = 8 μg mL−1), and spectinomycin (MIC = 32 μg mL−1) as potent direct antimicrobials.Spectrum of activity of AZT and spectinomycin was tested against a collection of 103 representative Enterobacteriaceae strains (≈50% CRE). AZT, a nucleoside analog used to treat human immunodeficiency virus, demonstrated an MIC50of 2 μg mL−1. Spectinomycin, an antibiotic used to treat gonorrhea, had an MIC50of 32 μg mL−1. Therefore, a significant percentage of CRE strains appeared relatively susceptible to these antimicrobials. These data identified AZT and spectinomycin as available agents warranting further study for potential treatment of multidrug-resistant CRE infection. Our results provide proof of principle and impetus for performing a large-scale HTS for discovery of novel, small-molecule adjunctives and antibacterial agents directly targeting CRE.
