title: High-resolution hyaluronidase inhibition profiling combined with HPLC-HRMS-SPE-NMR for identification of anti-necrosis constituents in Chinese plants used to treat snakebite.
abstract: Inhibition of the necrotizing hyaluronidase, phospholipase A2 and protease enzymes in four snake venoms by crude water and ethanol extracts of 88 plant species used against snakebites in traditional Chinese medicine was measured. High-resolution hyaluronidase inhibition profiles were constructed for the 22 plants showing highest hyaluronidase inhibition, and the results were used to guide subsequent structural analysis towards specific hyaluronidase inhibitors. Structural analysis was performed by high-performance liquid chromatography, high-resolution mass spectrometry, solid-phase extraction and nuclear magnetic resonance spectroscopy, i.e., HPLC-HRMS-SPE-NMR. This allowed identification of four non-tannin inhibitors, i.e., lansiumamide B (6) from Clausena excavata Burm.f., myricetin 3-O-β-D-glucopyranoside (7) from Androsace umbellata (Lour.) Merr., and vitexin (8) and 4',7-dihydroxy-5-methoxyflavone-8-C-β-D-glucopyranoside (9) from Oxalis corniculata L. Absolute configuration of 2,3-dihydroxy-N-methyl-3-phenyl-N-[(Z)-styryl]propanamide (1) was determined using the Mosher method, which revealed two enantiomers, i.e., (2S,3R)-2,3-dihydroxy-N-methyl-3-phenyl-N-[(Z)-styryl]propanamide and (2R,3S)-2,3-dihydroxy-N-methyl-3-phenyl-N-[(Z)-styryl]propanamide with a ratio of 7:3.
authors.affiliation: Department of Drug Design and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, Universitetsparken 2, DK-2100 Copenhagen, Denmark.
authors.forename: Yueqiu
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Department of Drug Design and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, Universitetsparken 2, DK-2100 Copenhagen, Denmark.
authors.forename: Dan
authors.initials: D
authors.lastname: Staerk
authors.affiliation: Department of Drug Design and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, Universitetsparken 2, DK-2100 Copenhagen, Denmark.
authors.forename: Mia N
authors.initials: MN
authors.lastname: Nielsen
authors.affiliation: Department of Drug Design and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, Universitetsparken 2, DK-2100 Copenhagen, Denmark.
authors.forename: Nils
authors.initials: N
authors.lastname: Nyberg
authors.affiliation: Department of Drug Design and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, Universitetsparken 2, DK-2100 Copenhagen, Denmark. Electronic address: anna.jager@sund.ku.dk.
authors.forename: Anna K
authors.initials: AK
authors.lastname: Jäger
mesh_terms: D047310:Apigenin / Q000302:isolation & purification; D002851:Chromatography, High Pressure Liquid; D065089:Glycoside Hydrolase Inhibitors / Q000737:chemistry; D006821:Hyaluronoglucosaminidase / Q000037:antagonists & inhibitors* / Q000378:metabolism; D008516:Medicine, Chinese Traditional; D015394:Molecular Structure; D019906:Nuclear Magnetic Resonance, Biomolecular; D065825:Oxalidaceae / Q000737:chemistry; D064801:Phospholipase A2 Inhibitors / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology*; D010936:Plant Extracts / Q000737:chemistry; D012909:Snake Bites / Q000188:drug therapy*; D052616:Solid Phase Extraction; D013634:Tannins / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology*
chemical_list: D065089:Glycoside Hydrolase Inhibitors; D064801:Phospholipase A2 Inhibitors; D010936:Plant Extracts; D013634:Tannins; D047310:Apigenin; C032731:vitexin; D006821:Hyaluronoglucosaminidase
keywords: Androsace umbellata; Clausena excavata; HPLC–HRMS–SPE–NMR; High-resolution profiling; Hyaluronidase inhibition; Oxalis corniculata; Snakebite