Structure-Activity Relationship of Sulfonyl Piperazine LpxH Inhibitors Analyzed by an LpxE-Coupled Malachite Green Assay.
Structure-Activity Relationship of Sulfonyl Piperazine LpxH Inhibitors Analyzed by an LpxE-Coupled Malachite Green Assay.
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DOI：
10.1021/acsinfecdis.8b00364
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发表时间：
2019-02
影响因子：
5.3
通讯作者：
Minhee Lee;Jinshi Zhao;Seung‐Hwa Kwak;J. Cho;M. Lee;R. Gillespie;D. Kwon;Hyunji Lee;Hyun-Ju Park;Qinglin Wu;P. Zhou;Jiyong Hong
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文献类型：
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作者：
Minhee Lee;Jinshi Zhao;Seung‐Hwa Kwak;J. Cho;M. Lee;R. Gillespie;D. Kwon;Hyunji Lee;Hyun-Ju Park;Qinglin Wu;P. Zhou;Jiyong Hong
在脂质A生物合成的Raetz途径中的UDP-2，3-二酰基葡糖胺焦磷酸酶LpxH是绝大多数革兰氏阴性病原体中的必需酶，并且是极好的新型抗生素靶标。32 P-放射自显影薄层色谱法已被广泛用于LpxH活性的分析，但它是不方便的评价大量的LpxH抑制剂在一个较长的时间段。在这里，我们报告了一个耦合，非放射性LpxH检测，利用最近发现的风产液菌脂质A 1-磷酸酶LpxE的定量去除的1-磷酸盐从脂质X，LpxH催化的产物;随后释放的无机磷酸盐定量的比色孔雀石绿色测定，允许监测的LpxH催化。使用这样一个耦合的酶法测定，我们报告了一系列的磺酰基哌嗪LpxH抑制剂的生化特性。我们的分析建立了这类化合物的初步结构-活性关系，并揭示了两个芳环，两个疏水基团和一个氢键受体的药效团。我们希望我们的研究结果将有助于开发更有效的LpxH抑制剂作为潜在的抗菌剂。
The UDP-2,3-diacylglucosamine pyrophosphatase LpxH in the Raetz pathway of lipid A biosynthesis is an essential enzyme in the vast majority of Gram-negative pathogens and an excellent novel antibiotic target. The 32P-radioautographic thin-layer chromatography assay has been widely used for analysis of LpxH activity, but it is inconvenient for evaluation of a large number of LpxH inhibitors over an extended time period. Here, we report a coupled, nonradioactive LpxH assay that utilizes the recently discovered Aquifex aeolicus lipid A 1-phosphatase LpxE for quantitative removal of the 1-phosphate from lipid X, the product of the LpxH catalysis; the released inorganic phosphate is subsequently quantified by the colorimetric malachite green assay, allowing the monitoring of the LpxH catalysis. Using such a coupled enzymatic assay, we report the biochemical characterization of a series of sulfonyl piperazine LpxH inhibitors. Our analysis establishes a preliminary structure-activity relationship for this class of compounds and reveals a pharmacophore of two aromatic rings, two hydrophobic groups, and one hydrogen-bond acceptor. We expect that our findings will facilitate the development of more effective LpxH inhibitors as potential antibacterial agents.