title: An analysis method for simultaneous screening of deoxyribonucleic acid-binding active compounds and investigating their mechanisms by ultra-fast liquid chromatography tandem mass spectrometry coupled with fluorescence detection technology.
abstract: DNA has been known as the cellular target for many cytotoxic anticancer agents over the years. Discovering DNA-binding compounds has become an active research area, while various DNA-binding mechanisms make the drug discovery even more difficult. In this article, we present a novel analysis method to rapidly identify specific DNA-binding compounds from Pyrrosia lingua (Thunb.) using DNA-dual-fluorescent probes, ethidium bromide and Hoechst 33258, with the technology of ultra-fast liquid chromatography-diode array detector-tandem mass spectrometry and dual-wavelength fluorescence detector (UFLC-DAD-MS(n)-DFLD). Sixty-two compounds were identified, of which 22 were found to be active in DNA-binding. After investigation of their dose-response behaviors and structure-activity relationships, chlorogenic acids and flavonoid glycosides were found to be DNA-binders via both minor groove-binding and intercalation modes. The precision, reproducibility and stability of this method were validated by vitexin. The established system was sensitive, precise, and reliable to be used for both screening of DNA-binding compounds and investigating of their mechanisms.
authors.affiliation: School of Pharmaceutical Sciences, Peking University, Beijing 100191, China. Electronic address: zcm899@163.com.
authors.forename: Cangman
authors.initials: C
authors.lastname: Zhang
authors.affiliation: School of Pharmaceutical Sciences, Peking University, Beijing 100191, China.
authors.forename: Biao
authors.initials: B
authors.lastname: Ren
authors.affiliation: Department of Pharmaceutical Sciences, University of Tennessee Health Science Center, Memphis, TN 38163, USA.
authors.forename: Zongtao
authors.initials: Z
authors.lastname: Lin
authors.affiliation: School of Pharmaceutical Sciences, Peking University, Beijing 100191, China.
authors.forename: Lingkun
authors.initials: L
authors.lastname: Tong
authors.affiliation: School of Pharmaceutical Sciences, Peking University, Beijing 100191, China. Electronic address: hw9505@bjmu.edu.cn.
authors.forename: Hong
authors.initials: H
authors.lastname: Wang
authors.affiliation: School of Pharmaceutical Sciences, Peking University, Beijing 100191, China. Electronic address: chenshizhong66@163.com.
authors.forename: Shizhong
authors.initials: S
authors.lastname: Chen
mesh_terms: D006690:Bisbenzimidazole / Q000737:chemistry; D002726:Chlorogenic Acid / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004247:DNA / Q000737:chemistry*; D004996:Ethidium / Q000737:chemistry*; D005419:Flavonoids / Q000737:chemistry; D005453:Fluorescence; D005456:Fluorescent Dyes / Q000737:chemistry; D006027:Glycosides / Q000737:chemistry; D007364:Intercalating Agents / Q000737:chemistry; D010936:Plant Extracts / Q000737:chemistry*; D018515:Plant Leaves / Q000737:chemistry; D029621:Polypodiaceae / Q000737:chemistry*; D015203:Reproducibility of Results; D013329:Structure-Activity Relationship; D053719:Tandem Mass Spectrometry / Q000379:methods
chemical_list: D005419:Flavonoids; D005456:Fluorescent Dyes; D006027:Glycosides; D007364:Intercalating Agents; D010936:Plant Extracts; D002726:Chlorogenic Acid; D004247:DNA; D004996:Ethidium; D006690:Bisbenzimidazole
keywords: DNA-binding; Ethidium bromide; Hoechst 33258; Pyrrosia lingua (Thunb.); UFLC-MS