Screening epidermal growth factor receptor antagonists from Radix et Rhizoma Asari by two-dimensional liquid chromatography.

Radix et Rhizoma Asari is a traditional Chinese medicine, and has many pharmacological effects, such as calming, analgesia, anti-inflammation, antiarrhythmic, antihypertensive, antivirus, etc. But few studies have screened the active compounds from extracts of Radix et Rhizoma Asari for tumor therapy. In this study, a two-dimensional liquid chromatography system was built to screen active compounds acting on epidermal growth factor receptor (EGFR) from Radix et Rhizoma Asari. The screening result showed that asarinin from Radix et Rhizoma Asari was the targeted component that could act on EGFR specificity. The competitive binding assay and molecular docking assay results showed asarinin binding with EGFR in similar manner as with gefitinib, which was used as a positive control drug. Then the antitumor effect of asarinin was studied through cell growth assay in vitro. The results showed that gefitinib and asarinin could inhibit highly expressed EGFR cell growth in a dose-dependent manner in the range of dose from 0.10 to 102.4 μM. This two-dimensional liquid chromatography system will be a useful method in drug discovery from natural medicinal herbs for searching potential antitumor candidates.

[1]  Langchong He,et al.  Investigation of calcium antagonist–L-type calcium channel interactions by a vascular smooth muscle cell membrane chromatography method , 2010, Analytical and bioanalytical chemistry.

[2]  R. Sheridan,et al.  New methods in computer-aided drug design , 1987 .

[3]  K. Gibson,et al.  ZD1839 (Iressa): an orally active inhibitor of epidermal growth factor signaling with potential for cancer therapy. , 2002, Cancer research.

[4]  Qinhua Chen,et al.  Screening active anti-breast cancer compounds from Cortex Magnolia officinalis by 2D LC-MS. , 2013, Journal of separation science.

[5]  C. Sherr,et al.  Principles of Tumor Suppression , 2004, Cell.

[6]  R. A. Shalliker,et al.  Comprehensive two-dimensional separations of complex mixtures using reversed-phase reversed-phase liquid chromatography. , 2004, Journal of chromatography. A.

[7]  Ruimiao Chang,et al.  Screening vasoconstriction inhibitors from traditional Chinese medicines using a vascular smooth muscle/cell membrane chromatography-offline-liquid chromatography-mass spectrometry. , 2011, Journal of separation science.

[8]  Jie Zhang,et al.  Applications of HPLC/MS in the analysis of traditional Chinese medicines , 2011, Journal of pharmaceutical analysis.

[9]  Establishment of A431 cell membrane chromatography-RPLC method for screening target components from Radix Caulophylli. , 2011, Journal of separation science.

[10]  Dwight R Stoll,et al.  Fast, comprehensive two-dimensional liquid chromatography. , 2007, Journal of chromatography. A.

[11]  N. Yang,et al.  Antiproliferative and antimetastatic effects of the ethanolic extract of Phellinus igniarius (Linnearus: Fries) Quelet. , 2008, Journal of ethnopharmacology.

[12]  J. Farndon,et al.  EPIDERMAL-GROWTH-FACTOR RECEPTOR STATUS AS PREDICTOR OF EARLY RECURRENCE OF AND DEATH FROM BREAST CANCER , 1987, The Lancet.

[13]  F. Regnier,et al.  Stoichiometric displacement of solvent by non-polar solutes in reversed-phase liquid chromatography. , 1985, Journal of chromatography.

[14]  Yanhong Song,et al.  High-expression β(1) adrenergic receptor/cell membrane chromatography method based on a target receptor to screen active ingredients from traditional Chinese medicines. , 2014, Journal of separation science.

[15]  Sicen Wang,et al.  Screening active components acting on α1A adrenergic receptors from agrimony using a Sprague-Dawley rat prostate cell membrane chromatography online coupled HPLC/MS method , 2012 .

[16]  Sicen Wang,et al.  Screening of target compounds from Fructus Piperis using high α₁A adrenoreceptor expression cell membrane chromatography online coupled with high performance liquid chromatography tandem mass spectrometry. , 2013, Journal of pharmaceutical and biomedical analysis.

[17]  C. James,et al.  Identical splicing of aberrant epidermal growth factor receptor transcripts from amplified rearranged genes in human glioblastomas. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[18]  Yanmin Zhang,et al.  A new A431/cell membrane chromatography and online high performance liquid chromatography/mass spectrometry method for screening epidermal growth factor receptor antagonists from Radix sophorae flavescentis. , 2010, Journal of chromatography. A.

[19]  Jae-Hoon Kim,et al.  Crystal Structure of the Complex of Human Epidermal Growth Factor and Receptor Extracellular Domains , 2002, Cell.

[20]  Jie Yu,et al.  Studies on morphology and aristolochic acid analogue constituents of Asarum campaniflorum and a comparison with two official species of Asari Radix et Rhizoma , 2010, Journal of Natural Medicines.

[21]  S. Shin,et al.  Rapid Mycobacterial Liquid Culture-Screening Method for Mycobacterium avium Complex Based on Secreted Antigen-Capture Enzyme-Linked Immunosorbent Assay , 2009, Clinical and Vaccine Immunology.

[22]  Sicen Wang,et al.  Screening target components from Radix salviae miltiorrhiae using an EGFR/CMC-online-HPLC/MS method , 2012 .

[23]  N. Saijo,et al.  Antitumor activity of the selective epidermal growth factor receptor‐tyrosine kinase inhibitor (EGFR‐TKI) Iressa® (ZD1839) in an EGFR‐expressing multidrug‐resistant cell line in vitro and in vivo , 2002, International journal of cancer.

[24]  Meng Sun,et al.  Simultaneous screening of four epidermal growth factor receptor antagonists from Curcuma longa via cell membrane chromatography online coupled with HPLC-MS. , 2013, Journal of separation science.

[25]  Shengli Han,et al.  Screening active compounds acting on the epidermal growth factor receptor from Radix scutellariae via cell membrane chromatography online coupled with HPLC/MS. , 2012, Journal of pharmaceutical and biomedical analysis.

[26]  Fanbo Jing,et al.  Design, synthesis and primary activity evaluation of l-arginine derivatives as amino-peptidase N/CD13 inhibitors , 2009, Bioorganic & Medicinal Chemistry.

[27]  S. Schreiber,et al.  Target-oriented and diversity-oriented organic synthesis in drug discovery. , 2000, Science.

[28]  Jennifer Griffiths,et al.  A brief history of mass spectrometry. , 2008, Analytical chemistry.

[29]  Ming-Jin Liang,et al.  Screening, analysis and in vitro vasodilatation of effective components from Ligusticum Chuanxiong. , 2005, Life sciences.