RESUMEN
A methanol extract of the leaves of Cephalotaxus harringtonia var. nana and its ethyl acetate (EtOAc)-soluble fraction demonstrated strong antitumor activity against A549 and HT-29 cell lines. The EtOAc-soluble fraction was purified by column chromatography and high-performance liquid chromatography (HPLC) using a reverse-phase column to yield three novel acyl flavonoids and a biflavonoid, along with 15 other known compounds that included flavonoids, biflavonoids, and other phenolics. The structures of the new compounds were elucidated using spectral data from HR-MS and NMR, including two-dimensional NMR studies, as (2R,3R)-3-O-eicosanoyltaxifolin (1), (2R,3R)-3-O-docosanoyltaxifolin (2), (2R,3R)-3-O-tetracosanoyltaxifolin (3), and 6-methyl-4',7,7â³-tri-O-methylamentoflavone (4). The isolated compounds, including the known compounds, were tested for possible antitumor activity; some of the biflavones were found to be active. The potent antitumor activity of the extract was attributed to Cephalotaxus alkaloids, such as homoharringtonine (20).
Asunto(s)
Productos Biológicos/química , Flavonoides/química , Harringtoninas/química , Cromatografía Líquida de Alta Presión , Humanos , Espectroscopía de Resonancia Magnética , Estructura Molecular , Hojas de la Planta/químicaRESUMEN
Lancemaside A, a triterpenoid saponin isolated from the roots of Codonopsis lanceolata, has been reported to ameliorate the reduction of blood testosterone levels induced by immobilization stress in mice. In the present study, we investigated the metabolism and absorption of lancemaside A in mice. After oral administration of lancemaside A at 100 mg/kg body weight, the unmetabolized compound appeared rapidly in plasma (t (max) = 0.5 h). Lancemaside A has a low bioavailability (1.1%) because of its metabolism by intestinal bacteria and its poor absorption in the gastrointestinal tract. Furthermore, we identified four metabolites from the cecum of mice after oral administration of lancemaside A: codonolaside II, echinocystic acid, echinocystic acid 28-O-beta-D: -xylopyranosyl-(1 --> 4)-alpha-L: -rhamnopyranosyl-(1 --> 2)-alpha-L: -arabinopyranosyl ester, and echinocystic acid 28-O-alpha-L: -rhamnopyranosyl-(1 --> 2)-alpha-L: -arabinopyranosyl ester. Among these metabolites, codonolaside II and echinocystic acid were detected in plasma, and their t (max) values were 4 and 8 h, respectively. These findings should be helpful for understanding the mechanism of the biological effect of lancemaside A.
Asunto(s)
Codonopsis/química , Raíces de Plantas/química , Saponinas/metabolismo , Administración Oral , Animales , Ciego/química , Ciego/metabolismo , Cromatografía Liquida , Ésteres/química , Ésteres/metabolismo , Masculino , Espectrometría de Masas , Ratones , Estructura Molecular , Ácido Oleanólico/análogos & derivados , Ácido Oleanólico/química , Ácido Oleanólico/metabolismo , Saponinas/sangre , Saponinas/química , Saponinas/farmacocinéticaRESUMEN
PURPOSE: This observational case series investigated the occurrence and distribution of proptosis in Japanese patients with dysthyroid ophthalmopathy (DO). The relationship among lid retraction, lid swelling, and enlargement of extraocular muscles was also analyzed. METHODS: From 1993 to 2002, 10 931 patients (2240 men and 8691 women, average age; 39.9 ± 14.0 years) with abnormal thyroid function were examined for proptosis. Proptosis was measured using a Hertel exophthalmometer, and lid swelling and retraction were classified. Orbital magnetic resonance imaging or computed tomography was used to quantify enlargement of the extraocular muscles. RESULTS: Clinically significant proptosis (≥15 mm) was present in 74.2% patients and was classified as mild (15-17 mm, 39.1%), moderate (18-20 mm, 25.9%), or severe (>20 mm, 9.2%). The average exophthalmometer reading was 17.2 ± 3.2 mm, with proptosis less pronounced with age (p < 0.001) and more pronounced in men than women (18.2 ± 3.4 mm vs. 17.0 ± 3.1 mm; p < 0.001). Lid retraction was present in 57.7% of patients, lid swelling in 46.9%, and muscle enlargement in 40.8% of patients. Multiple regression analysis revealed proptosis was strongly correlated with a young age and the male gender, followed by extraocular muscle enlargement and lid retraction and swelling (p < 0.001). CONCLUSIONS: This study assessed the measurable features of proptosis in Japanese patients with DO and contributes to the understanding by correlating symptoms and signs of DO.
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Exoftalmia/complicaciones , Oftalmopatía de Graves/complicaciones , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Pueblo Asiatico , Niño , Preescolar , Técnicas de Diagnóstico Oftalmológico , Exoftalmia/diagnóstico , Enfermedades de los Párpados/diagnóstico , Femenino , Oftalmopatía de Graves/diagnóstico , Humanos , Hipertrofia , Imagen por Resonancia Magnética , Masculino , Persona de Mediana Edad , Músculos Oculomotores/patología , Tomografía Computarizada por Rayos X , Adulto JovenRESUMEN
We developed a rapid and simple analytical method for the simultaneous determination of seven 3,28-bidesmosidic triterpenoid saponins in the roots of Codonopsis lanceolata. The saponins are lancemaside A, lancemaside B, lancemaside C, lancemaside E, lancemaside G, foetidissimoside A, and aster saponin Hb. Root samples were extracted with 50% methanol and prepared for analysis. Saponins were detected by reversed-phase high-performance liquid chromatography with electrospray ionization mass spectrometry, and ginsenoside Rb(1) was used as an internal standard. The overall recoveries of all saponins were 92-116%, and the relative standard deviation values of intra- and inter-day precision were lower than 3.7 and 7.7%, respectively. Eight root samples collected from Korea and Japan were analyzed using the developed method. Lancemaside A was the most abundant saponin in the root samples from Korea, ranging from 2.65 to 3.64 mg/g dry root. However, the maximum content of lancemaside A among Japanese samples was 0.101 mg/g dry root.
Asunto(s)
Cromatografía Líquida de Alta Presión/métodos , Codonopsis/química , Raíces de Plantas/química , Saponinas/análisis , Espectrometría de Masa por Ionización de Electrospray/métodos , Estructura Molecular , Reproducibilidad de los Resultados , Saponinas/química , Triterpenos/análisis , Triterpenos/químicaRESUMEN
Liquid chromatography coupled with sequential mass spectrometry (LC-MS(n)) has been used to identify 3,28-bidesmosidic triterpenoid saponins, lancemaside A (1), foetidissimoside A (2), aster saponin Hb (3), lancemaside E (4), lancemaside B (5), lancemaside F (6), lancemaside G (7), lancemaside C (8), and lancemaside D (9) in the roots of Codonopsis lanceolata. Structural information about both the aglycone and the sugar moiety at the C-3 position of saponins was obtained in the negative-ion mode. On the other hand, positive-ion spectra mainly provide structural information about the sugar chains of saponins, especially the oligosaccharide moiety at the C-28 position. During subsequent fragmentation of the product ions derived from the oligosaccharide moiety at the C-28 position, fragments produced by sequential loss of a monosaccharide unit were observed. Furthermore, the structural features of two unknown saponins in the roots of C. lanceolata were assigned on the basis of the fragmentation patterns of the known saponins. These studies demonstrate that LC-MS(n) analysis has great potential for the identification and characterization of triterpenoid saponins in plant extracts.
Asunto(s)
Codonopsis/química , Saponinas/aislamiento & purificación , Triterpenos/aislamiento & purificación , Cromatografía Liquida/métodos , Espectrometría de Masas/métodos , Raíces de Plantas , Saponinas/química , Triterpenos/químicaRESUMEN
INTRODUCTION: Lancemaside A is a saponin that inhibits decreases in blood testosterone level and thus prevents or ameliorates symptoms associated with male climacteric disorder. Our initial attempt to preparative isolation of lancemaside A from the saponin fraction of Codonopsis lanceolata roots by a preparative HPLC did not give a clear result. OBJECTIVE: To develop a simple and efficient method for the preparative isolation of lancemaside A from the hot water extract of C. lanceolata roots using centrifugal partition chromatography (CPC). METHODOLOGY: The saponin fraction obtained from the hot water extract of C. lanceolata roots was used as the sample for preparative-scale separation of lancemasides by CPC using n-hexane:n-butanol:methanol:0.1% aqueous formic acid (3:4:1:6, v/v) as the two-phase solvent system. The upper phase (organic phase) of the two-phase solvent system was used as the mobile phase, and 0.5 g of saponin fraction was applied for separation by CPC. Each fraction that was separated by CPC was analysed by HPLC, and the fractions containing each of the separated compounds were pooled together, and then were purified by simple preparative HPLC. RESULTS: The demonstrated separation sequence, hot water extraction, DIAION HP-20 column chromatography, CPC and preparative HPLC, yielded lancemaside A, foetidissimoside A and astersaponin Hb in their pure forms. CONCLUSION: The simple and efficient method for the preparative isolation of lancemaside A along with two other saponins, foetidissimoside A and astersaponin Hb, from the saponin fraction of C. lanceolata was established using CPC.
Asunto(s)
Cromatografía Líquida de Alta Presión/métodos , Codonopsis/química , Saponinas/aislamiento & purificación , Espectroscopía de Resonancia Magnética , Extractos Vegetales/química , Espectrofotometría UltravioletaRESUMEN
In the course of the development of new designer foods using the roots of Codonopsis lanceolata, we found that hot-water extracts of C. lanceolata recovered decreased testosterone levels in the blood and accelerated the restoration of reproductive dysfunction induced by hyperthermic treatment in male mice. Thus we studied the constituents of the polar fraction of the roots of C. lanceolata and identified six new triterpene saponins, lancemasides B (2), C (3), D (4), E (5), F (6), and G (7), along with the known saponin lancemasaide A (1) and phenylpropanoid glycosides 8-10. The structures of the new compounds 2-7 were determined by means of spectral data including 2D-NMR studies and chemical reactions to be oleanan-type bisdesmoside with sugars at C-3 and C-28. Compounds 2-6 have echinocystic acid as an aglycone, and compound 7 has asterogenic acid as an aglycone. Identification of the sugars and determination of their D,L-chiralities were carried out by application of the exciton chirality method to the per-O-p-bromobenzoylmethyl sugar derived from saponins.