Magnetic beads-based neuraminidase enzyme microreactor as a drug discovery tool for screening inhibitors from compound libraries and fishing ligands from natural products.
Magnetic beads-based neuraminidase enzyme microreactor as a drug discovery tool for screening inhibitors from compound libraries and fishing ligands from natural products.
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基于磁珠的神经氨酸酶微反应器作为药物发现工具，用于从化合物库中筛选抑制剂和从天然产物中捕获配体
DOI：
10.1016/j.chroma.2018.07.031
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发表时间：
2018-09-21
期刊：
影响因子：
--
通讯作者：
Jiang ZJ
中科院分区：
文献类型：
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作者：
Zhao YM;Wang LH;Luo SF;Wang QQ;Moaddel R;Zhang TT;Jiang ZJ
神经氨酸酶（NA）是一种糖苷水解酶，已被提议作为流感的潜在治疗靶点。因此，鉴定调节NA活性的化合物可能具有重要的治疗意义。本研究的目的是开发一种药物发现工具，用于从化合物库和天然植物提取物中鉴定新型NA调节剂。将NA固定到磁珠表面，并表征了NA功能化磁珠的固有催化活性。以酶活性（水解率）为指标，对12种植物次生代谢产物的抑制活性进行了筛选，确定了黄酮类化合物具有较好的抗NA活性。使用由奥司他韦、石蒜碱和苦参碱组成的人工测试混合物优化用固定化酶的配体钓鱼，然后用红花的粗提取物进行概念验证实验。结合HPLC-MS/MS技术，初步鉴定了木犀草素-7-O-β-D-葡萄糖苷、木犀草素、3，5-二-O-咖啡酰奎尼酸和3，4-二-O-咖啡酰奎尼酸为紫花苜蓿中的神经氨酸酶抑制剂配体。这是关于使用神经氨酸酶功能化磁珠从植物基质中识别活性配体的第一份报告，它为从复杂的生物混合物中重新识别NA调节剂奠定了基础。
Neuraminidase (NA) is a glycoside hydrolase that has been proposed as a potential therapeutic target for influenza. Thus, the identification of compounds that modulate NA activity could be of great therapeutic importance. The aim of this study is to develop a drug discovery tool for the identification of novel modulators of NA from both compound libraries and natural plant extracts. NA was immobilized onto the surface of magnetic beads and the inherent catalytic activity of NA-functionalized magnetic beads was characterized. Based on the enzymatic activity (hydrolysis ratio), the inhibitory activities of 12 compounds from plant secondary metabolites were screened, and the desired anti-NA activities of flavonoids were certified. Ligand fishing with the immobilized enzyme was optimized using an artificial test mixture consisting of oseltamivir, lycorine and matrine prior to carrying out the proof-of-concept experiment with the crude extract of Flos Lonicerae. The combination of ligand fishing and HPLC-MS/MS identified luteolin-7-O-β-D-glucoside, luteolin, 3,5-di-O-caffeoylquinic acid and 3,4-di-O-caffeoylquinic acid as neuraminidase inhibitory ligands in Flos Lonicerae. This is the first report on the use of neuraminidase functionalized magnetic beads for the identification of active ligands from a botanical matrix, and it sets the basis for the de novo identification of NA modulators from complex biological mixtures.