Institute of Traditional Chinese Medicine & Natural Products, College of Pharmacy/Guangdong Province Key Laboratory of Pharmacodynamic Constituents of TCM and New Drugs Research/International Cooperative Laboratory of Traditional Chinese Medicine Modernization and Innovative Drug Development of Ministry of Education (MOE) of China, Jinan University, Guangzhou 510632, China.
Division of BioAnalytical Chemistry, Amsterdam Institute of Molecules, Medicines and Systems, Vrije Universiteit Amsterdam, 1081 HV Amsterdam, The Netherlands.
Molecules. 2022 Oct 15;27(20):6923. doi: 10.3390/molecules27206923.
How to rapidly and accurately screen bioactive components from complex natural products remains a major challenge. In this study, a screening platform for pancreatic lipase (PL) inhibitors was established by combining magnetic beads-based ligand fishing and high-resolution bioassay profiling. This platform was well validated using a mixture of standard compounds, i.e., (-)- epigallocatechin gallate (EGCG), luteolin and schisandrin. The dose-effect relationship of high-resolution bioassay profiling was demonstrated by the standard mixture with different concentrations for each compound. The screening of PL inhibitors from green tea extract at the concentrations of 0.2, 0.5 and 1.0 mg/mL by independent high-resolution bioassay profiling was performed. After sample pre-treatment by ligand fishing, green tea extract at the concentration of 0.2 mg/mL was specifically enriched and simplified, and consequently screened through the high-resolution bioassay profiling. As a result, three PL inhibitors, i.e., EGCG, (-)-Gallocatechin gallate (GCG) and (-)-Epicatechin gallate (ECG), were rapidly identified from the complex matrix. The established platform proved to be capable of enriching affinity binders and eliminating nonbinders in sample pre-treatment by ligand fishing, which overcame the technical challenges of high-resolution bioassay profiling in the aspects of sensitivity and resolution. Meanwhile, the high-resolution bioassay profiling possesses the ability of direct bioactive assessment, parallel structural analysis and identification after separation. The established platform allowed more accurate and rapid screening of PL inhibitors, which greatly facilitated natural product-based drug screening.
如何快速、准确地从复杂天然产物中筛选生物活性成分仍然是一个主要挑战。在本研究中，通过将基于磁珠的配体钓捕与高分辨生物测定谱相结合，建立了一种胰脂肪酶（PL）抑制剂的筛选平台。该平台通过混合标准化合物（即表没食子儿茶素没食子酸酯（EGCG）、木樨草素和五味子醇甲）进行了很好的验证。通过对具有不同浓度的每种化合物的标准混合物，证明了高分辨生物测定谱的剂量效应关系。通过独立的高分辨生物测定谱，在 0.2、0.5 和 1.0 mg/mL 浓度下从绿茶提取物中筛选 PL 抑制剂。在通过配体钓捕进行样品预处理后，在 0.2 mg/mL 的浓度下，绿茶提取物被特异性地浓缩和简化，然后通过高分辨生物测定谱进行筛选。结果，从复杂基质中快速鉴定出三种 PL 抑制剂，即 EGCG、(-)-Gallocatechin gallate (GCG) 和 (-)-Epicatechin gallate (ECG)。所建立的平台证明能够在配体钓捕的样品预处理中富集亲和结合物并消除非结合物，从而克服了高分辨生物测定谱在灵敏度和分辨率方面的技术挑战。同时，高分辨生物测定谱具有直接生物活性评估、平行结构分析和分离后的鉴定能力。所建立的平台允许更准确、快速地筛选 PL 抑制剂，从而极大地促进了基于天然产物的药物筛选。
