Quality Evaluation and Characterization of Fractions with Biological Activity from Ephedra Herb Extract and Ephedrine Alkaloids-Free Ephedra Herb Extract

• Published in: Chemical & pharmaceutical bulletin Vol. 68; no. 2; pp. 140 - 149
• Main Authors: Yoshimura, Morio, Amakura, Yoshiaki, Hyuga, Sumiko, Hyuga, Masashi, Nakamori, Shunsuke, Maruyama, Takuro, Oshima, Naohiro, Uchiyama, Nahoko, Yang, Jinwei, Oka, Hideki, Ito, Hideyuki, Kobayashi, Yoshinori, Odaguchi, Hiroshi, Hakamatsuka, Takashi, Hanawa, Toshihiko, Goda, Yukihiro
• Format: Journal Article
• Language: English
• Published: Japan The Pharmaceutical Society of Japan 01.02.2020; Japan Science and Technology Agency
• Subjects: Alkaloids; Alkaloids - chemistry; Alkaloids - pharmacology; analgesia; Analgesics; Analgesics - chemistry; Analgesics - pharmacology; Animals; anti-influenza; Antiviral Agents - chemistry; Antiviral Agents - pharmacology; Bans; Biflavonoids - chemistry; Biflavonoids - pharmacology; Biological activity; c-Met; Catechin - chemistry; Catechin - pharmacology; Catechol; Cell Line, Tumor; Chromatography; Column chromatography; condensed tannin; Decomposition reactions; Dogs; Ephedra; Ephedra - chemistry; Ephedrine; Ephedrine - chemistry; Ephedrine - pharmacology; ephedrine alkaloids-free Ephedra Herb extract (EFE); gel permeation chromatography (GPC); Herbs; High performance liquid chromatography; Humans; Influenza; Kinases; Liquid chromatography; Madin Darby Canine Kidney Cells; Male; Mice; Molecular weight; NMR; Nuclear magnetic resonance; Plant Extracts - chemistry; Plant Extracts - pharmacology; Polystyrene; Polystyrene resins; Proanthocyanidins - chemistry; Proanthocyanidins - pharmacology; Protein Kinase Inhibitors - chemistry; Protein Kinase Inhibitors - pharmacology; Proto-Oncogene Proteins c-met - antagonists & inhibitors; Pyrogallol; Quality assessment; Tannins; gel permeation chromatography (GPC); condensed tannin; anti-influenza; c-Met; ephedrine alkaloids-free Ephedra Herb extract (EFE); analgesia
• ISSN: 0009-2363; 1347-5223
• DOI: 10.1248/cpb.c19-00761

Previously, we reported that the c-Met inhibitory effect of Ephedra Herb extract (EHE) is derived from ingredients besides ephedrine alkaloids. Moreover, analgesic and anti-influenza activities of EHE and ephedrine alkaloids-free Ephedra Herb extract (EFE) have been reported recently. In this study, we examined the fractions containing c-Met kinase inhibitory activity from EHE and the fractions with analgesic and anti-influenza activities from EFE, and elucidated the structural characteristics of the active fractions. Significant c-Met kinase activity was observed in 30, 40, and 50% methanol (MeOH) eluate fractions obtained from water extract of EHE using Diaion HP-20 column chromatography. Similarly, 20 and 40% MeOH, and MeOH eluate fractions obtained from water extract of EFE were found to display analgesic and anti-influenza activities. Reversed phase-HPLC analysis of the active fractions commonly showed broad peaks characteristic of high-molecular mass condensed tannin. The active fractions were analyzed using 13C-NMR and decomposition reactions; the deduced structures of active components were high-molecular mass condensed tannins, which were mainly procyanidin B-type and partly procyanidin A-type, including pyrogallol- and catechol-type flavan 3-ols as extension and terminal units. HPLC and gel permeation chromatography (GPC) analyses estimated that the ratio of pyrogallol- and catechol-type was approximately 9 : 2, and the weight-average molecular weight based on the polystyrene standard was >45000. Furthermore, GPC-based analysis was proposed as the quality evaluation method for high-molecular mass condensed tannin in EHE and EFE.