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Planta Med 2021; 87(05): 375-382
DOI: 10.1055/a-1272-3997
Biological and Pharmacological Activity
Original Papers

Rapid Screening for EGFR Inhibitor in Rhei Radix et Rhizoma by HTRF Assay Coupled with HPLC Peak Fractionation

Yu Fu‡
1   School of Pharmacy, Henan University of Chinese Medicine, Zhengzhou, Henan, China
3   Collaborative Innovation Center for Respiratory Disease Diagnosis and Treatment & Chinese Medicine Development of Henan Province, Henan University of Chinese Medicine, Zhengzhou, Henan, China
,
Zhishen Xie‡
2   Center for Scientific Research, Henan University of Chinese Medicine, Zhengzhou, Henan, China
,
Peng Zhao
3   Collaborative Innovation Center for Respiratory Disease Diagnosis and Treatment & Chinese Medicine Development of Henan Province, Henan University of Chinese Medicine, Zhengzhou, Henan, China
,
Shuangshuang Lv
1   School of Pharmacy, Henan University of Chinese Medicine, Zhengzhou, Henan, China
,
Suiqing Chen 
1   School of Pharmacy, Henan University of Chinese Medicine, Zhengzhou, Henan, China
3   Collaborative Innovation Center for Respiratory Disease Diagnosis and Treatment & Chinese Medicine Development of Henan Province, Henan University of Chinese Medicine, Zhengzhou, Henan, China
› Author AffiliationsSupported by: National Natural Science Foundation of China 81703680
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Abstract

In this paper, an HPLC peak fractionation approach combined with homogeneous time-resolved fluorescence analysis is proposed for screening epidermal growth factor receptor inhibitors from Rhei Radix et Rhizoma. With this approach, the amount of sample used for a single HPLC run is sufficient for performing a multiple assay due to the miniaturization ability of the homogeneous time-resolved fluorescence technology. This allows for improving the stability and repeatability of the activity assay for each fraction. From a total of 26 fractions collected from the Rhei Radix et Rhizoma extract, 13 fractions exhibit inhibitory activity against the epidermal growth factor receptor. The structures of activity compounds were determined by HPLC-LTQ-Orbitrap MS, revealing the presence of gallic acid, rhein, and emodin with IC50 values of 21.5, 5.29, and 10.2 µM, respectively. The ligand epidermal growth factor receptor interactions were explored by molecular docking simulations, and the inhibitory effects of the three compounds on A549 cell growth were tested in vitro by an MTT assay. This study demonstrates the suitability of the present screening method for drug discovery in natural products.

Key words

Peak fractionation - HTRF - EGFR - Rheum palmatum - Polygonaceae

Supporting Information

    The purity of each peak collected from HPLC and profiles of RRR are available as Supporting Information.

  • Supporting Information


Publication History

Received: 05 April 2020

Accepted after revision: 21 September 2020

Article published online:
11 November 2020

© 2020. Thieme. All rights reserved.

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