RESUMO
Excessive nitric oxide (NO) produced by inducible NO synthase (iNOS) acts as a causative regulator in various inflammatory disease states. Carpesium divaricatum has been used in Korean traditional herbal medicine for its antipyretic, analgesic, vermifugic, and anti-inflammatory properties. We investigated the molecular mechanism for the suppression of lipopolysaccharide/interferon-gamma (LPS/IFN-gamma)-induced NO production in RAW 264.7 macrophages by the sesquiterpene lactone 2beta,5-epoxy-5,10-dihydroxy-6alpha-angeloyloxy-9beta-isobutyloxy-germacran-8alpha,12-olide (C-1), which has been identified recently as a new compound from C. divaricatum. C-1 decreased NO production in LPS/IFN-gamma-stimulated RAW 264.7 cells in a concentration-dependent manner, with an IC50 of approximately 2.16 microM; however, it had no direct effect on the iNOS activity of fully LPS/IFN-gamma-stimulated RAW 264.7 cells. Furthermore, treatment with C-1 led to a decrease in iNOS protein and mRNA. These effects appear to be due to inhibition of nuclear factor-kappaB (NF-kappaB) activation through a mechanism involving stabilization of the NF-kappaB/inhibitor of the kappaB (I-kappaB) complex, since inhibition of NF-kappaB DNA binding activity by C-1 was accompanied by a parallel reduction of nuclear translocation of subunit p65 of NF-kappaB and I-kappaBalpha degradation. Taken together, the results suggest that the ability of C-1 to inhibit iNOS gene expression may be responsible, in part, for its anti-inflammatory effects.
Assuntos
Proteínas I-kappa B , NF-kappa B/antagonistas & inibidores , Óxido Nítrico Sintase/biossíntese , Plantas Medicinais/química , Sesquiterpenos/farmacologia , Animais , Transporte Biológico/efeitos dos fármacos , Catálise , Núcleo Celular/efeitos dos fármacos , Núcleo Celular/metabolismo , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Proteínas de Ligação a DNA/metabolismo , Repressão Enzimática/efeitos dos fármacos , Interferon gama/farmacologia , Ligases/metabolismo , Lipopolissacarídeos/farmacologia , Camundongos , Inibidor de NF-kappaB alfa , NF-kappa B/metabolismo , Óxido Nítrico/metabolismo , Óxido Nítrico Sintase/genética , Óxido Nítrico Sintase/metabolismo , Óxido Nítrico Sintase Tipo II , Nitritos/metabolismo , Fitoterapia , RNA Mensageiro/biossíntese , RNA Mensageiro/efeitos dos fármacosRESUMO
A new cyanogenic glycoside, sutherlandin-5-trans-p-coumarate was isolated along with a known cardiosdiospermin-5-(4-hydroxy) benzoate fr the aerial parts of Sorbaria sorbifolia (L.) A. Br. var. stellipila MAX. (Rosaceae). The structure of the new compound was established based on spectral evidence.
Assuntos
Ácidos Cumáricos/isolamento & purificação , Glicosídeos/isolamento & purificação , Plantas Medicinais/química , China , Cromatografia por Troca Iônica , Ácidos Cumáricos/química , Glicosídeos/química , Espectroscopia de Ressonância Magnética , Espectrometria de Massas de Bombardeamento Rápido de Átomos , Espectrofotometria Infravermelho , Espectrofotometria UltravioletaRESUMO
We investigated the mechanism of suppression of inducible nitric oxide synthase (iNOS) by two beta-carboline alkaloids isolated from Melia azedarach, 4,8-dimethoxy-1-vinyl-beta-carboline (compound 1, C-1) and 4-methoxy-1-vinyl-beta-carboline (compound 2, C-2). iNOS activity in a cell-free extract of lipopolysaccharide/interferon-gamma-stimulated RAW 264.7 cells was found to be markedly increased, and this increase was prevented by C-1 and C-2, accompanied by the parallel reduction in nitrite accumulation in culture medium. However, C-1 and C-2 had no further effect on the iNOS activity prepared from fully lipopolysaccharide/interferon-gamma-stimulated RAW 264.7 cells. Treatment with C-1 or C-2 decreased the levels of iNOS protein and mRNA in a concentration-dependent manner. In addition, prostaglandin E(2) production, cyclooxygenase-2 protein and DNA binding of nuclear factor-kappaB (NF-kappaB) in lipopolysaccharide-stimulated RAW 264.7 cells were reduced by these compounds. These results indicate that C-1 and C-2 primarily inhibit iNOS and cyclooxygenase-2 activities via the suppression of de novo synthesis of these two enzymes, and that the inhibition of iNOS expression may be associated with the inhibition of NF-kappaB activation. Taken together, the results suggest that suppression of iNOS and cyclooxygenase-2 induction by lipopolysaccharide is responsible for the anti-inflammatory activity of these alkaloids through selective inhibition of the expression of genes, which play important roles in inflammatory signaling pathways.
Assuntos
Alcaloides/farmacologia , Carbolinas/farmacologia , Inibidores Enzimáticos/farmacologia , Óxido Nítrico Sintase/antagonistas & inibidores , Animais , Linhagem Celular , Citrulina/biossíntese , Ciclo-Oxigenase 2 , Dinoprostona/biossíntese , Isoenzimas/efeitos dos fármacos , Isoenzimas/genética , Lipopolissacarídeos/farmacologia , Macrófagos/enzimologia , Camundongos , NF-kappa B/efeitos dos fármacos , NF-kappa B/metabolismo , Óxido Nítrico Sintase/genética , Óxido Nítrico Sintase/metabolismo , Óxido Nítrico Sintase Tipo II , Nitritos/metabolismo , Prostaglandina-Endoperóxido Sintases/efeitos dos fármacos , Prostaglandina-Endoperóxido Sintases/genéticaRESUMO
In bioassay-guided search for inducible nitric oxide synthase (iNOS) inhibitory compounds from higher plants of South Korea, two beta-carboline alkaloids, 4-methoxy-1-vinyl-beta-carboline (1) and 4,8-dimethoxy-l-vinyl-beta-carboline (2) have been isolated from the cortex of Melia azedarach var. japonica. The structures of these compounds were elucidated on the basis of spectroscopic data. Compounds 1 and 2 showed marked inhibitory activity of iNOS on LPS- and interferon-gamma-stimulated RAW 264.7 cells.
Assuntos
Carbolinas/farmacologia , Inibidores Enzimáticos/farmacologia , Óxido Nítrico Sintase/antagonistas & inibidores , Plantas Medicinais/química , Carbolinas/isolamento & purificação , Linhagem Celular , Cromatografia por Troca Iônica , Relação Dose-Resposta a Droga , Inibidores Enzimáticos/isolamento & purificação , Humanos , Interferon gama/farmacologia , Coreia (Geográfico) , Lipopolissacarídeos/farmacologia , Espectroscopia de Ressonância Magnética , Óxido Nítrico Sintase Tipo II , Extratos Vegetais/química , Extratos Vegetais/farmacologia , Espectrofotometria Infravermelho , Espectrofotometria UltravioletaRESUMO
Two new furanolignans (3, 5), together with three known lignans (1, 2, 4), were isolated from the stem of Lindera obtusiloba (Lauraceae). The structures of the compounds were determined as actifolin (1), pluviatilol (2), 5,6-dihydroxymatairesinol (3), (+)-syringaresinol (4), and (+)-9'-O-trans-feruloyl-5,5'-dimethoxylariciresinol (5) on the basis of physicochemical and spectroscopic evidences. Compounds 1, 2, 3, and 5 showed cytotoxicity against a small panel of human tumor cell lines with ED50 values of 3.40 to approximately 19.27 microg/ml.
Assuntos
Antineoplásicos Fitogênicos/isolamento & purificação , Lignanas/química , Plantas Medicinais/química , Antineoplásicos Fitogênicos/farmacologia , Cromatografia em Gel , Cromatografia Líquida de Alta Pressão , Cromatografia em Camada Fina , Ensaios de Seleção de Medicamentos Antitumorais , Humanos , Coreia (Geográfico) , Lignanas/isolamento & purificação , Espectroscopia de Ressonância Magnética , Espectrometria de Massas , Espectrofotometria InfravermelhoRESUMO
A new acyclic diterpene (1) and a known acyclic diterpene, 12(S)-hydroxygeranylgeraniol (2) were isolated from the aerial parts of Carpesium divaricatum. The structure of 1 was determined to be (2E,10E)-1,12-dihydroxy-18-acetoxy-3,7,15-trimethylhexadeca- 2,10,14-triene (1) on the basis of spectroscopic studies. Compounds 1 and 2 exhibited cytotoxicity against cultured human tumor cell lines, A549, SK-OV-3, SK-MEL-2, XF498, and HCT15, with ED50 values ranging from 4.3-10.2 micrograms/ml and 4.1-8.3 micrograms/ml, respectively.
Assuntos
Antineoplásicos Fitogênicos/isolamento & purificação , Plantas Medicinais/química , Antineoplásicos Fitogênicos/química , Antineoplásicos Fitogênicos/farmacologia , Humanos , Espectroscopia de Ressonância Magnética , Células Tumorais CultivadasRESUMO
In a bioassay-guided search for anti-allergic compounds from higher plants of Korea, polymethoxyflavones, 3',4',5,6,7,8-hexamethoxyflavone (1), 5-hydroxy-3',4',6,7,8-pentamethoxyflavone (II) and 3',4',5,7,8,-pentamethoxyflavone (III) have been isolated from the immature peels of Citrus unshiu. Structures of these compounds were elucidated on the basis of spectroscopic techniques. Compounds I and II inhibited dose-dependently histamine release from the rat peritoneal mast cells activated by compound 48/80 or anti-DNP IgE.
Assuntos
Antialérgicos/farmacologia , Citrus/química , Liberação de Histamina/efeitos dos fármacos , Extratos Vegetais/farmacologia , Animais , Antialérgicos/isolamento & purificação , Relação Dose-Resposta a Droga , Coreia (Geográfico) , Extratos Vegetais/isolamento & purificação , Ratos , Ratos Wistar , Espectroscopia de Infravermelho com Transformada de FourierRESUMO
Two new monogalactosylmonoacylglycerols ( 1, 2) and two known compounds ( 3, 4) were isolated from Hydrocotyle ramiflora. Based on physicochemical evidence and spectral data, the structures of 1, 2, 3, and 4 were determined as (2 S)-1- O-(7 Z,10 Z,13 Z-hexadecatrienoyl)-3- O-beta-galactopyranosylglycerol, (2 S)-1- O-(9 Z,12 Z-octadecadienoly)-3- O-beta-galactopyranosyl-glycerol, alpha-spinasterol, and capsidiol 3-acetate, respectively.
RESUMO
To overcome multidrug resistance (MDR) in cancer chemotherapy, we prepared various plant extracts and searched for a component which is effective for inhibition of MDR. MDR inhibition activity was determined by measuring cytotoxicity to MDR cells using multidrug resistant human fibrocarcinoma KB V20C, which is resistant to 20 nM vincristine and expresses high level of mdr1 gene. Of various plant extracts, the MeOH extract of the root of Aconitum pseudo-laeve var. erectum was found to have potent inhibitory activity on MDR. The bioassay-guided fractionation of the MeOH extract of the plant led to the isolation of an alkaloid, lycaconitine, as an active principle. And the IC50 of lycaconitine for KB V20C cells was 74 micrograms/ml.
Assuntos
Alcaloides/isolamento & purificação , Antineoplásicos Fitogênicos/isolamento & purificação , Antineoplásicos/farmacologia , Resistência a Múltiplos Medicamentos/fisiologia , Plantas Medicinais/química , Alcaloides/química , Alcaloides/farmacologia , Antineoplásicos Fitogênicos/química , Antineoplásicos Fitogênicos/farmacologia , Sobrevivência Celular/efeitos dos fármacos , Humanos , Células Tumorais CultivadasRESUMO
A new epoxynaphthoquinol derivative, 3-acetyl-2-methyl-1, 5-dihydroxy-2,3-epoxynaphthoquinol (I), was isolated from the root of Rumex japonicus. The structure was elucidated by high field 1D and 2D NMR techniques.
Assuntos
Polygonaceae/química , Espectroscopia de Ressonância Magnética , Estrutura Molecular , Raízes de Plantas/químicaRESUMO
Four thymol derivatives, 2,5-dimethoxythymol (1), 2-methoxythymol isobutyrate (2), 10-isobutyloxy-8,9-epoxythymolisobutyrate (3) and 10-(2-methylbutyloxy)-8,9-epoxythymolisobutyrate (4) were isolated from the aerial parts of Carpesium divaricatum. The structures were elucidated by high field 1D and 2D NMR techniques.
Assuntos
Asteraceae/química , Timol/análogos & derivados , Espectroscopia de Ressonância Magnética , Espectrofotometria Infravermelho , Espectrofotometria Ultravioleta , Timol/química , Timol/isolamento & purificaçãoRESUMO
In a bioassay-guided search for cytotoxic compounds from higher plants of South Korea, four new sesquiterpenes of the germacranolide type, named cardivins A (1), B (2), C (3), and D (4), have been isolated from the aerial parts of Carpesium divaricatum. Structures of these compounds were elucidated on the basis of spectroscopic techniques. Compounds 1, 2, 3, and 4 showed cytotoxicity to the human tumor cells, A-549 (nonsmall cell lung), SK-OV-3 (ovary), SK-MEL-2 (skin), XF-498 (central nervous system), and HCT-15 (colon).
Assuntos
Antineoplásicos Fitogênicos/isolamento & purificação , Plantas Medicinais/química , Sesquiterpenos/isolamento & purificação , Antineoplásicos Fitogênicos/farmacologia , Ensaios de Seleção de Medicamentos Antitumorais , Humanos , Coreia (Geográfico) , Espectroscopia de Ressonância Magnética , Espectrometria de Massas , Sesquiterpenos/farmacologia , Espectrofotometria Infravermelho , Células Tumorais CultivadasRESUMO
A new coumarin-monoterpene ether, artekeiskeanin A (1) and two known coumarins, dracunculin (2) and scopoletin (3) were isolated from the aerial parts of Artemisia keiskeana. The structure of 1 was determined to be 7-(trans-8-oxogeranyloxy)-6-methoxycoumarin on the basis of spectroscopic studies.
Assuntos
Cumarínicos/química , Plantas Medicinais , Terpenos/química , Cumarínicos/isolamento & purificação , Medicamentos de Ervas Chinesas , Coreia (Geográfico) , Estrutura Molecular , Terpenos/isolamento & purificaçãoRESUMO
Bioassay-guided fractionation of the aerial parts ofPilea mongolica (Urticaceae) afforded two cytotoxic triterpenoids, epi-oleanolic acid (I) and oxo-oleanolic acid (II). The structures of the compounds were confirmed by spectral and synthetic evidences. CompoundI and compoundII exhibited cytotoxicity against cultured human tumor cell lines, A549 (non small cell lung adenocarcinoma), SK-OV-3 (ovarian), SK-MEL-2 (skin melanoma), XF498 (CNS) and HCT15 (colon) with ED(50) values of 3.2 approximately 8.1 mug/ml and 0.7 approximately 6.8 mug/ml, respectively.
RESUMO
The acivity-guided fractionation upon the MeOH extract of the aerial parts ofSorbaria sorbifolia var.stellipila led to the isolation of two cucurbitacin-compounds, cucurbitacin D and cucurbitacin F, as active principles. Two compounds were shown to exhibit significant cytotoxicity against cultured human tumor cell lines, A-549, SK-OV-3, SK-MEL-2, XF-498, and HCT 15.
RESUMO
A high-performance liquid chromatographic method with column switching has been developed for the simultaneous determination of cefamandole and cefamandole nafate in plasma and urine. The plasma and urine samples were injected onto a precolumn packed with Corasil RP C18 (37-50 microns) after simple dilution with an internal standard solution in 0.05 M phosphoric acid. Polar plasma and urine components were washed out using 0.05 M phosphoric acid. After valve switching, the concentrated drugs were desorbed in back-flush mode and separated by a reversed-phase C8 column with methanol-5 mM tetrabutylammonium bromide (45:55, v/v) as the mobile phase. The method showed excellent precision with good sensitivity and speed, and a detection limit of 0.5 microgram/ml. The total analysis time per sample was less than 30 min, and the mean coefficients of variation for intra- and inter-assay were both less than 4.9%. The method has been successfully applied to plasma and urine samples for human volunteers after intravenous injection of cefamandole nafate.
Assuntos
Cefamandol/análogos & derivados , Cefamandol/análise , Cefamandol/sangue , Cefamandol/urina , Cromatografia Líquida de Alta Pressão/instrumentação , Humanos , Reprodutibilidade dos TestesRESUMO
For routine analysis of diclofenac sodium in plasma, a new high performance liquid chromatographic method, which is combined with column-switching technique is developed. The precolumn packed with Corasil RP C-18 was connected to analytical column by switching system in order to enrich the sample drugs in plasma without extraction. This method showed excellent sensitivity, precision and reproducibility. The limit of detection, using a 100 microL injection of plasma, was 0.1 micrograms/mL and the mean coefficient of variation for intra- and inter-assay was better than 4.6%. Total analysis time was 20 min between injections. The present method offers distinct practical advantages over conventional liquid-liquid extraction methods of sample preparation with respect to time, effort, recovery, and sample volume required. The method has been applied to the samples from rats receiving oral administration of diclofenac sodium.