Identification of natural compound inhibitors for multidrug efflux pumps of Escherichia coli and Pseudomonas aeruginosa using in silico high-throughput virtual screening and in vitro validation.
Identification of natural compound inhibitors for multidrug efflux pumps of Escherichia coli and Pseudomonas aeruginosa using in silico high-throughput virtual screening and in vitro validation.
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DOI：
10.1371/journal.pone.0101840
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Hopper W
中科院分区：
文献类型：
--
作者：
Aparna V;Dineshkumar K;Mohanalakshmi N;Velmurugan D;Hopper W
铜绿假单胞菌和大肠杆菌对多种抗生素具有耐药性，使得感染的治疗非常困难。阻力的主要机制是外排泵的功能。 MexAB-OprM 和 AcrAB-TolC 是三联外排泵组件，分别负责铜绿假单胞菌和大肠杆菌的多重耐药性。预计更容易流出的底物具有共同的药效团特征图。本研究采用了排除具有类似外排底物特征的化合物的新标准，从而细化了选择过程并丰富了抑制剂的识别过程。使用针对 AcrB 和 MexB 蛋白的高通量虚拟筛选创建和筛选植物化学物质的内部数据库，并通过与使用已知外排底物生成的常见药效团模型（AADHR、ADHNR、AAHNR、AADHN、AADNR、AAADN、AAADR、AAANR、AAAHN、AAADD 和 AAADH）匹配进行过滤。与任何一种或多种外排底物模型匹配的植物化学命中被排除在研究之外。使用 XP 对接对 AcrB 和 MexB 蛋白对接不具有与外排底物模型相似的特征的命中。 XP 对接的最佳命中通过棋盘协同测定和溴化乙锭积累测定对其外排抑制效力进行了验证。根据协同潜力过滤毛花苷 C 和二黄酮，并使用溴化乙锭积累研究验证其外排抑制效力。这些化合物表现出增加细菌细胞内溴化乙锭积累的能力，如化合物存在时荧光的增加所证明的那样。由于计算机筛选与体外阳性外排抑制活性之间存在良好的相关性，毛花苷 C 和二黄酮这两种化合物可能成为有前景的外排泵抑制剂，并可有效用于针对铜绿假单胞菌和大肠杆菌耐药菌株的联合治疗。
Pseudomonas aeruginosa and Escherichia coli are resistant to wide range of antibiotics rendering the treatment of infections very difficult. A main mechanism attributed to the resistance is the function of efflux pumps. MexAB-OprM and AcrAB-TolC are the tripartite efflux pump assemblies, responsible for multidrug resistance in P. aeruginosa and E. coli respectively. Substrates that are more susceptible for efflux are predicted to have a common pharmacophore feature map. In this study, a new criterion of excluding compounds with efflux substrate-like features was used, thereby refining the selection process and enriching the inhibitor identification process. An in-house database of phytochemicals was created and screened using high-throughput virtual screening against AcrB and MexB proteins and filtered by matching with the common pharmacophore models (AADHR, ADHNR, AAHNR, AADHN, AADNR, AAADN, AAADR, AAANR, AAAHN, AAADD and AAADH) generated using known efflux substrates. Phytochemical hits that matched with any one or more of the efflux substrate models were excluded from the study. Hits that do not have features similar to the efflux substrate models were docked using XP docking against the AcrB and MexB proteins. The best hits of the XP docking were validated by checkerboard synergy assay and ethidium bromide accumulation assay for their efflux inhibition potency. Lanatoside C and diadzein were filtered based on the synergistic potential and validated for their efflux inhibition potency using ethidium bromide accumulation study. These compounds exhibited the ability to increase the accumulation of ethidium bromide inside the bacterial cell as evidenced by these increase in fluorescence in the presence of the compounds. With this good correlation between in silico screening and positive efflux inhibitory activity in vitro, the two compounds, lanatoside C and diadzein could be promising efflux pump inhibitors and effective to use in combination therapy against drug resistant strains of P. aeruginosa and E. coli.
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影响因子：
3.2
作者：
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通讯作者：
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影响因子：
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作者：
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DOI：
10.3402/ljm.v6i0.5870
发表时间：
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期刊：
The Libyan journal of medicine
影响因子：
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通讯作者：
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DOI：
10.1016/j.ijantimicag.2009.06.007
发表时间：
2009-10-01
影响因子：
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作者：
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影响因子：
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作者：
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