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Trace component fishing strategy based on offline two-dimensional liquid chromatography combined with PRDX3-surface plasmon resonance for Uncaria alkaloids

文献类型：期刊论文

作者Ni, Hui2,3; Zhang, Zijia3; Lu, Ye4; Liu, Yaowen2,3; Zhou, Yang3; Wu, Wenyong3; Kong, Xinqin1,3; Shen, Liling1,3; Chen, Sihan1,3; Long, Huali3
刊名JOURNAL OF PHARMACEUTICAL ANALYSIS
出版日期2025-09-01
卷号15期号:9页码:13
关键词Offline two-dimensional liquid chromatography Peroxiredoxin 3 Surface plasmon resonance Trace component fishing strategy Uncaria alkaloids
ISSN号2095-1779
DOI10.1016/j.jpha.2025.101244
英文摘要The rapid screening of bioactive constituents within traditional Chinese medicine (TCM) presents a significant challenge to researchers. Prevailing strategies for the screening of active components in TCM often overlook trace components owing to their concealment by more abundant constituents. To address this limitation, a fishing strategy based on offline two-dimensional liquid chromatography (2D-LC) combined with surface plasmon resonance (SPR) was utilized to screen bioactive trace components targeting peroxiredoxin 3 (PRDX3), using Uncaria alkaloids (UAs) as a case study. Initially, an orthogonal preparative offline 2D-LC system combining a positively charged C18 column and a conventional C18 column under disparate mobile phase conditions was constructed. To fully reveal the trace alkaloids, 13 2D fractions of UAs were prepared, and their components were characterized using mass spectrometry (MS). Subsequently, employing PRDX3 as the targeting protein, a SPR-based screening approach was established and rigorously validated with geissoschizine methyl ether (GSM) serving as a positive control for binding. Employing this refined strategy, 29 candidate binding alkaloids were fished from the 13 2D fractions. Notably, combining offline 2D-LC with SPR increased the yield of candidate binding components from 10 to 29 when compared to SPR-based screening alone. Subsequent binding affinity assays confirmed that PRDX3 was a direct binding target for the 12 fished alkaloids, with isovallesiachotamine (IV), corynoxeine N-oxide (CO-N), and cadambine (CAD) demonstrating the highest affinity for PRDX3. Their interactions were further validated through molecular docking analysis. Subsequent intracellular H2O2 measurement assays and transfection experiments confirmed that these three trace alkaloids enhanced PRDX3-mediated H2O2 clearance. In conclusion, this study introduced an innovative strategy for the identification of active trace components in TCM. This approach holds promise for accelerating research on medicinal components within this field. (c) 2025 The Authors. Published by Elsevier B.V. on behalf of Xi'an Jiaotong University. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
WOS关键词FLIGHT MASS-SPECTROMETRY ; PREPARATIVE ISOLATION ; IDENTIFICATION
资助项目National Key Research and Development Program of China[2022YFC3501704] ; National Key Research and Development Program of China[2023YFC3504205] ; Sanming Project of Medicine in Shenzhen, China[ZZYSM202106004]
WOS研究方向Pharmacology & Pharmacy
语种英语
WOS记录号WOS:001577737300001
出版者ELSEVIER
源URL[http://119.78.100.183/handle/2S10ELR8/321519]  
专题国家级研究中心_原创新药研究全国重点实验室
通讯作者Zhang, Hao; Hou, Jinjun; Wu, Wanying
作者单位1.Univ Chinese Acad Sci, Beijing 100049, Peoples R China
2.Nanjing Univ Chinese Med, Sch Chinese Mat Med, Nanjing 210029, Peoples R China
3.Chinese Acad Sci, Shanghai Inst Mat Med, Natl Engn Res Ctr TCM Standardizat Technol, Shanghai Res Ctr Modernizat Tradit Chinese Med, Shanghai 201203, Peoples R China
4.Shanghai Univ Tradit Chinese Med, Inst Interdisciplinary Integrat Med Res, Shanghai 201203, Peoples R China
5.Chinese Acad Sci, Shanghai Inst Mat Med, Chem Biol Res Ctr, State Key Lab Drug Res, Shanghai 201203, Peoples R China
推荐引用方式
GB/T 7714		 Ni, Hui,Zhang, Zijia,Lu, Ye,et al. Trace component fishing strategy based on offline two-dimensional liquid chromatography combined with PRDX3-surface plasmon resonance for Uncaria alkaloids[J]. JOURNAL OF PHARMACEUTICAL ANALYSIS,2025,15(9):13.
APA Ni, Hui.,Zhang, Zijia.,Lu, Ye.,Liu, Yaowen.,Zhou, Yang.,...&Wu, Wanying.(2025).Trace component fishing strategy based on offline two-dimensional liquid chromatography combined with PRDX3-surface plasmon resonance for Uncaria alkaloids.JOURNAL OF PHARMACEUTICAL ANALYSIS,15(9),13.
MLA Ni, Hui,et al."Trace component fishing strategy based on offline two-dimensional liquid chromatography combined with PRDX3-surface plasmon resonance for Uncaria alkaloids".JOURNAL OF PHARMACEUTICAL ANALYSIS 15.9(2025):13.

入库方式： OAI收割

来源：上海药物研究所

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