Discovery of a novel inhibitor of NAD(P)(+)-dependent malic enzyme (ME2) by high-throughput screening
文献类型：期刊论文
| 作者 | Wen, Yi1; Xu, Lei2; Chen, Fang-lei2; Gao, Jing1; Li, Jing-ya2 ; Hu, Li-hong2; Li, Jia2
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| 刊名 | ACTA PHARMACOLOGICA SINICA
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| 出版日期 | 2014-05 |
| 卷号 | 35期号:5页码:674-684 |
| 关键词 | malic enzyme ME2 inhibitor natural product NPD389 high-throughput screening structure-activity relationship enzyme kinetics thermal shift assay |
| ISSN号 | 1671-4083 |
| DOI | 10.1038/aps.2013.189 |
| 文献子类 | Article |
| 英文摘要 | Aim: Malic enzymes are oxidative decarboxylases with NAD(+) or NAD(P)(+) as cofactor that catalyze the conversion of L-malate to pyruvate and CO2. The aim of this study was to discover and characterize a potent inhibitor of human NAD(P)(+)-dependent malic enzyme 2 (ME2). Methods: Recombinant human ME2-His-Tag fusion protein was overexpressed in E coli and purified with Ni-NTA resin. A high-throughput screening (HTS) assay was developed to find ME2 inhibitors. Detergent Brij-35 was used to exclude false positives. The characteristics of the inhibitor were analyzed with enzyme kinetics analysis. A thermal shift assay for ME2 was carried out to verify the binding of the inhibitor with the enzyme. Results: An HTS system for discovering ME2 inhibitors was established with a Z' factor value of 0.775 and a signal-to-noise ratio (S/N) of 9.80. A library containing 12 683 natural products was screened. From 47 hits, NPD387 was identified as an inhibitor of ME2. The primary structure-activity relationship study on NPD387 derivatives showed that one derivative NPD389 was more potent than the parent compound NPD387 (the IC50 of NPD389 was 4.63 +/- 0.36 mu mol/L or 5.59 +/- 0.38 mu mol/L, respectively, in the absence or presence of 0.01% Brij-35 in the assay system). The enzyme kinetics analysis showed that NPD389 was a fast-binding uncompetitive inhibitor with respect to the substrate NAD(+) and a mixed-type inhibitor with respect to the substrate L-malate. Conclusion: NPD389 is a potent ME2 inhibitor that binds to the enzyme in a fast-binding mode, acting as an uncompetitive inhibitor with respect to the substrate NAD(+) and a mixed-type inhibitor with respect to the substrate L-malate. |
| WOS关键词 | THERMAL SHIFT ASSAYS ; STATISTICAL PARAMETER ; QUALITY-CONTROL ; DRUG DISCOVERY ; RNAI SCREENS ; PROGRESSION ; MECHANISM ; HIT ; ATP |
| 资助项目 | National Natural Science Foundation of China[81125023] ; National Natural Science Foundation of China[81021062] ; National Science and Technology Major Projects for Major New Drugs Innovation and Development[2013ZX09507002] ; Shanghai Commission of Science and Technology[11DZ2292200] ; Shanghai Commission of Science and Technology[13DZ2290300] |
| WOS研究方向 | Chemistry ; Pharmacology & Pharmacy |
| 语种 | 英语 |
| CSCD记录号 | CSCD:5142658 |
| WOS记录号 | WOS:000335447000012 |
| 出版者 | ACTA PHARMACOLOGICA SINICA |
| 源URL | [http://119.78.100.183/handle/2S10ELR8/277101]  |
| 专题 | 国家新药筛选中心 中科院受体结构与功能重点实验室 新药研究国家重点实验室 药物安全性评价中心 |
| 通讯作者 | Li, Jia |
| 作者单位 | 1.Jiangsu Univ, Sch Pharm, Zhenjiang 212013, Peoples R China; 2.Chinese Acad Sci, State Key Lab Drug Res, Shanghai Inst Mat Med, Shanghai 201203, Peoples R China |
| 推荐引用方式 GB/T 7714 | Wen, Yi,Xu, Lei,Chen, Fang-lei,et al. Discovery of a novel inhibitor of NAD(P)(+)-dependent malic enzyme (ME2) by high-throughput screening[J]. ACTA PHARMACOLOGICA SINICA,2014,35(5):674-684. |
| APA | Wen, Yi.,Xu, Lei.,Chen, Fang-lei.,Gao, Jing.,Li, Jing-ya.,...&Li, Jia.(2014).Discovery of a novel inhibitor of NAD(P)(+)-dependent malic enzyme (ME2) by high-throughput screening.ACTA PHARMACOLOGICA SINICA,35(5),674-684. |
| MLA | Wen, Yi,et al."Discovery of a novel inhibitor of NAD(P)(+)-dependent malic enzyme (ME2) by high-throughput screening".ACTA PHARMACOLOGICA SINICA 35.5(2014):674-684. |
入库方式： OAI收割
来源：上海药物研究所
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