RESUMO
Oat is the nutritious crop containing various compounds with antioxidant properties, such as polyphenols. In this study, we investigated the effect of germination and ultrafiltration process on polyphenol and avenanthramide contents in oat as well as their cytoprotective effect. Germination of oat for 48 hr significantly increased avenanthramide (5.5 to 11.3 mg/g) and polyphenol (115 to 155 mg GAE/g) contents. The compounds were more concentrated after ultrafiltration using 10 kDa membranes (polyphenol, 206 GAE/g; avenanthramide, 18 mg/g). In addition, oat extracts significantly reduced the cellular ROS level against tert-butyl hydroperoxide (t-BHP) stimulation in HepG2 cells. In the mechanistic study, oat extracts induced Nrf2 translocation to the nucleus by inhibition of Keap1 expression, resulting into upregulation of γ-GCS and NQO1. In conclusion, oat germination and ultrafiltration processes increased the polyphenol content, including that of avenanthramide. These extracts protected cells from t-BHP by radical scavenging activities and induced Nrf2 pathway activation. PRACTICAL APPLICATIONS: This study presents the method for avenanthramide-concentrated extract which is unique bioactive compounds in oat. In addition, antioxidant activity and their mechanisms of the avenanthramide-enriched extracts were evaluated. The polyphenol compounds including avenanthramide were found to increase after germination and ultrafiltration, thereby improving the radical scavenging ability. These results can be utilized as data for the development of health-promoting materials using oats.
Assuntos
Avena/crescimento & desenvolvimento , Extratos Vegetais/farmacologia , Polifenóis/análise , Polifenóis/farmacologia , ortoaminobenzoatos/farmacologia , Avena/química , Avena/genética , Avena/metabolismo , Germinação , Células Hep G2 , Humanos , Proteína 1 Associada a ECH Semelhante a Kelch/genética , Proteína 1 Associada a ECH Semelhante a Kelch/metabolismo , Fator 2 Relacionado a NF-E2/genética , Fator 2 Relacionado a NF-E2/metabolismo , Extratos Vegetais/análise , Extratos Vegetais/isolamento & purificação , Extratos Vegetais/metabolismo , Polifenóis/isolamento & purificação , Polifenóis/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Sementes/química , Sementes/genética , Sementes/crescimento & desenvolvimento , Sementes/metabolismo , Ultrafiltração , ortoaminobenzoatos/análise , ortoaminobenzoatos/isolamento & purificação , ortoaminobenzoatos/metabolismoRESUMO
A chitosan membrane composed by 60% (w/w) chitosan and 40% (w/w) Aliquat®336 has been proposed as a new biopolymeric support for electromembrane extraction. The new support has been characterized by Scanning Electron Microscopy, resulting a 30-35⯵m thickness. Amoxicillin, nicotinic acid, hippuric acid, salicylic acid, anthranilic acid, ketoprofen, naproxen and ibuprofen have been successfully extracted using the proposed support. Better enrichment factors were obtained for the acidic polar analytes than for the non-steroidal anti-inflammatory compounds (ranging from 118 for hippuric acid and 20 for ibuprofen). Electromembrane extraction was developed applying a DC voltage of 100â¯V, 1-octanol as supported liquid membrane and 20â¯min of extraction. The target analytes have also been satisfactorily extracted from human urine samples, providing high extraction efficiencies. The chitosan membrane is presented as a promising alternative for supporting liquid membrane compared to commonly used materials for this purpose.
Assuntos
Biopolímeros/química , Quitosana , Técnicas Eletroquímicas , Amoxicilina/química , Amoxicilina/isolamento & purificação , Hipuratos/química , Hipuratos/isolamento & purificação , Humanos , Ibuprofeno/química , Ibuprofeno/isolamento & purificação , Cetoprofeno/química , Cetoprofeno/isolamento & purificação , Naproxeno/química , Naproxeno/isolamento & purificação , Niacina/química , Niacina/isolamento & purificação , Ácido Salicílico/química , Ácido Salicílico/isolamento & purificação , ortoaminobenzoatos/química , ortoaminobenzoatos/isolamento & purificaçãoRESUMO
Leaves of the plant species Isatis indigotica Fortune ex Lindl. (Chinese woad) produce the metabolites tryptanthrin, indirubin and N-formylanthranilic acid upon spraying with an aqueous solution of copper chloride but not after spraying with water. The antifungal activities of these metabolites against the phytopathogens Alternaria brassicicola, Leptosphaeria maculans and Sclerotinia sclerotiorum established that tryptanthrin is a much stronger growth inhibitor of L. maculans than the phytoalexin camalexin. The biosynthetic precursors of tryptanthrin and N-formylanthranilic acid are proposed based on the deuterium incorporations of isotopically labeled compounds. The overall results suggest that tryptanthrin is a phytoalexin and indirubin and N-formylanthranilic acid are phytoanticipins in the plant species I. indigotica and that chemical diversity and biodiversity are intimately connected.
Assuntos
Alternaria/efeitos dos fármacos , Antifúngicos/farmacologia , Ascomicetos/efeitos dos fármacos , Medicamentos de Ervas Chinesas/farmacologia , Isatis/química , Quinazolinas/farmacologia , ortoaminobenzoatos/farmacologia , Antifúngicos/química , Antifúngicos/isolamento & purificação , Relação Dose-Resposta a Droga , Medicamentos de Ervas Chinesas/química , Medicamentos de Ervas Chinesas/isolamento & purificação , Indóis/química , Indóis/isolamento & purificação , Indóis/farmacologia , Testes de Sensibilidade Microbiana , Estrutura Molecular , Quinazolinas/química , Quinazolinas/isolamento & purificação , Relação Estrutura-Atividade , ortoaminobenzoatos/química , ortoaminobenzoatos/isolamento & purificaçãoRESUMO
Three new derivatives, anthranosides A-C (1-3), were discovered from a marine sponge-derived actinomycete Streptomyces sp. CMN-62. Their structures including absolute configurations were elucidated using MS and NMR spectroscopic data, X-ray single-crystal diffraction analysis, and chemical synthesis. Compounds 1 and 2 are epimers composed by linking the anthranilate unit to the fructofuranose moiety via a carbon-nitrogen bond, while 3 possessed a unique indole-containing scaffold. All compounds were tested for cytotoxicity, anti-influenza H1N1 virus, and NFκB inhibitory activities, and 3 showed anti-influenza activity. A possible Amadori rearrangement-based biosynthetic pathway was proposed to generate compounds 1-3.
Assuntos
Poríferos/microbiologia , Streptomyces/química , ortoaminobenzoatos/química , Animais , Vírus da Influenza A Subtipo H1N1/efeitos dos fármacos , NF-kappa B/antagonistas & inibidores , Streptomyces/fisiologia , ortoaminobenzoatos/isolamento & purificação , ortoaminobenzoatos/farmacologiaRESUMO
Avenanthramides (Avns), polyphenols found exclusively in oats, are emerging as promising therapeutic candidates for the treatment of several human diseases, including colon cancer. By engineering a Saccharomyces cerevisiae strain, we previously produced two novel phenolic compounds, N-(E)-p-coumaroyl-3-hydroxyanthranilic acid (Yeast avenanthramide I, YAvnI) and N-(E)-caffeoyl-3-hydroxyanthranilic acid (Yeast avenanthramide II, YAvnII), which are endowed with a structural similarity to bioactive oat avenanthramides and stronger antioxidant properties. In this study, we evaluated the ability of these yeast-derived recombinant avenanthramides to inhibit major hallmarks of colon cancer cells, including sustained proliferation, migration and epithelial-mesenchymal transition (EMT). Using the human colon adenocarcinoma cell line HT29, we compared the impact of YAvns and natural Avns, including Avn-A and Avn-C, on colon cancer cells by performing MTT, clonogenic, adhesion, migration, and anchorage-independent growth assays, and analyzing the expression of EMT markers. We found that both YAvns and Avns were able to inhibit colon cancer cell growth by increasing the expression of p21, p27 and p53 proteins. However, YAvns resulted more effective than natural compounds in inhibiting cancer cell migration and reverting major molecular features of the EMT process, including the down-regulation of E-cadherin mRNA and protein levels.
Assuntos
Adenocarcinoma/tratamento farmacológico , Antineoplásicos/farmacologia , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Neoplasias Colorretais/tratamento farmacológico , Transição Epitelial-Mesenquimal/efeitos dos fármacos , Saccharomyces cerevisiae/metabolismo , ortoaminobenzoatos/farmacologia , Adenocarcinoma/metabolismo , Adenocarcinoma/patologia , Antineoplásicos/isolamento & purificação , Neoplasias Colorretais/metabolismo , Neoplasias Colorretais/patologia , Inibidor de Quinase Dependente de Ciclina p21/metabolismo , Inibidor de Quinase Dependente de Ciclina p27/metabolismo , Células HT29 , Humanos , Invasividade Neoplásica , Transdução de Sinais/efeitos dos fármacos , Proteína Supressora de Tumor p53/metabolismo , ortoaminobenzoatos/isolamento & purificaçãoRESUMO
Seven new N-acylanthranilic acid derivatives (1-3, 4a/4b, 5a/5b) including two pairs of enantiomers (4a/4b and 5a/5b) were isolated from the leaves of Isatis indigotica Fortune. Their chemical structures were elucidated by extensive spectroscopic data analyses. The absolute configurations of compounds 4a/4b and 5a/5b were determined by comparison of the experimental and calculated ECD spectra. All compounds were tested for their anti-Aß1-42 aggregation activity. As a result, compounds 1 (72.1%), 2 (79.8%) and 5a (81.8%) showed stronger inhibitory activity than the positive control curcumin (67.0%). By the comparison between 5a (81.8%) and 5b (63.1%), it was found that stereochemical configurations may affect Aß1-42 aggregation activity, which was discussed through the molecular docking results of compounds 5a and 5b.
Assuntos
Peptídeos beta-Amiloides/antagonistas & inibidores , Isatis/química , Fragmentos de Peptídeos/antagonistas & inibidores , Folhas de Planta/química , ortoaminobenzoatos/isolamento & purificação , Simulação de Acoplamento Molecular , EstereoisomerismoRESUMO
Seven new anthranilamide derivatives, acoapetalines A-G (1-7), including one new naturally occurring compound (7), together with six known ones (8-13) were isolated from the whole plants of Aconitum apetalum. Their structures were established on the basis of extensive spectroscopic analyses (IR, UV, HRESIMS, 1D and 2D NMR). All compounds were evaluated for their anti-tobacco mosaic virus (anti-TMV) activities. The results showed that compound 10 exhibited significant anti-TMV activity with an inhibition rate of 67.57% (100⯵g/mL), which was higher than that of ningnamycin (52.89%).
Assuntos
Aconitum/química , Antivirais/isolamento & purificação , Vírus do Mosaico do Tabaco/efeitos dos fármacos , ortoaminobenzoatos/isolamento & purificação , Antivirais/farmacologia , Estrutura Molecular , Doenças das Plantas/prevenção & controle , Doenças das Plantas/virologia , Nicotiana/virologia , ortoaminobenzoatos/farmacologiaRESUMO
Four new 5-hydroxyanthranilic acid related compounds, named anthocidins Aâ»D (1â»4), two known analogues n-lauryl 5-hydroxyanthranilate (5) and isolauryl 5-hydroxyanthranilate (6), together with benzamide (7), 3-hydroxy-4-methoxycinnamamide (8), and (3S-cis)-hexahydro-3-[(3,4-dihydroxyphenyl)methyl]pyrrolo[1,2-a]pyrazine-1,4-dione (9), were isolated from the fermentation broth of the marine-derived actinomycete, Streptomyces sp. HDa1, which was isolated from the gut of a sea urchin, Anthocidaris crassispina, collected from Hainan Island, China. The structures of these secondary metabolites were elucidated on the basis of their 1D and 2D-NMR and mass spectroscopic data, and anthocidin A was confirmed by single-crystal X-ray diffraction with Cu Kα radiation. Anthocidins Aâ»D (1â»4) feature an acetyl group substitution at the amino group and varying alkyl side chains at the carboxyl group of 5-hydroxyanthranilic acid, and compound 5 was isolated as a natural product for the first time. The cytotoxic and antibacterial activity of compounds 1â»9 were evaluated.
Assuntos
Actinobacteria/patogenicidade , Antibacterianos/isolamento & purificação , Ouriços-do-Mar/microbiologia , Streptomyces/patogenicidade , ortoaminobenzoatos/isolamento & purificação , Actinobacteria/química , Animais , Antibacterianos/química , Antibacterianos/farmacologia , Linhagem Celular Tumoral , Sobrevivência Celular , China , Cristalografia por Raios X , Fermentação , Modelos Moleculares , Estrutura Molecular , Streptomyces/química , ortoaminobenzoatos/química , ortoaminobenzoatos/farmacologiaRESUMO
Avenanthramides are phenolic compounds found only in oats, and are of interest due to suggested bioactivities, including anti-inflammatory effects and induction of apoptosis. The objective of this work was to optimise a high performance liquid chromatography (HPLC) method for analysis of avenanthramides in food, and analyse the avenanthramide content in 45 oat fractions and products. The optimised HPLC method was based on triplicate extraction of 100â¯mg sample with 1â¯ml 80% ethanol in phosphate buffer (pH 2.8) and used gallacetophenone as an internal standard. Avenalumic acid-derived avenanthramide homologues 2fd and 2pd were also present, making up to 20% of the total avenanthramides detected in oats. The amounts of avenanthramides detected in oat products was 2-82⯵g/g. It was estimated that mean avenanthramide intake among oat consumers ranges from 0.3 to 2.1â¯mg/day, considerably lower than the amount used in studies that have investigated biological effects of avenanthramides in humans.
Assuntos
Avena/química , Cromatografia Líquida de Alta Pressão/métodos , ortoaminobenzoatos/análise , Ingestão de Alimentos , Grão Comestível/química , Feminino , Farinha/análise , Humanos , Masculino , Reprodutibilidade dos Testes , Suécia , Estados Unidos , ortoaminobenzoatos/isolamento & purificação , ortoaminobenzoatos/metabolismoRESUMO
N-glycan profiling is commonly accomplished by the derivatization of the enzymatically released oligosaccharides with a fluorophore, thereby facilitating their analysis by hydrophilic-interaction liquid chromatography (HILIC). These fluorescent dyes are often present in large excess during derivatization reactions, and their removal is typically required to minimize chromatographic interference. Herein, we report a reactivity-driven 2-phase extraction protocol with the aldehyde reagent octanal, which demonstrated efficient 2-aminobenzamide cleanup as well as high derivatized N-glycan recovery. This cleanup method can be performed within minutes, and therefore provides an alternative sample preparation route for N-glycan profiling with improved time efficiency and operational simplicity.
Assuntos
Oligossacarídeos/química , Oligossacarídeos/isolamento & purificação , ortoaminobenzoatos/química , ortoaminobenzoatos/isolamento & purificação , Cromatografia Líquida de Alta Pressão , Corantes Fluorescentes/química , Interações Hidrofóbicas e Hidrofílicas , Estrutura MolecularRESUMO
The use of polymer inclusion membranes (PIMs) as support of 1-octanol liquid membrane in electromembrane extraction (EME) procedure is proposed. Synthesis of PIMs were optimized to a composition of 29% (w/w) of cellulose triacetate as base polymer and 71% (w/w) of Aliquat®336 as cationic carrier. Flat PIMs of 25µm thickness and 6mm diameter were used. EME protocol was implemented for the simultaneous extraction of four non-steroidal anti-inflammatory drugs (NSAIDs) (salicylic acid, ketoprofen, naproxen and ibuprofen) and four highly polar acidic drugs (anthranilic acid, nicotinic acid, amoxicillin and hippuric acid). Posterior HPLC separation of the extracted analytes was developed with diode array detection. Recoveries in the 81-34% range were obtained. EME procedure was applied to human urine samples.
Assuntos
Anti-Inflamatórios não Esteroides/isolamento & purificação , Celulose/análogos & derivados , Técnicas Eletroquímicas , Compostos de Amônio Quaternário/química , Amoxicilina/isolamento & purificação , Amoxicilina/urina , Anti-Inflamatórios não Esteroides/urina , Celulose/química , Hipuratos/isolamento & purificação , Hipuratos/urina , Ibuprofeno/isolamento & purificação , Ibuprofeno/urina , Cetoprofeno/isolamento & purificação , Cetoprofeno/urina , Membranas Artificiais , Naproxeno/isolamento & purificação , Naproxeno/urina , Niacina/isolamento & purificação , Niacina/urina , Ácido Salicílico/isolamento & purificação , Ácido Salicílico/urina , ortoaminobenzoatos/isolamento & purificação , ortoaminobenzoatos/urinaRESUMO
A new alkaloid, 1,2-dihydrophenopyrrozin (1), along with five known compounds (2-6) was isolated from an axenic culture of the endophytic fungus, Bionectria sp., obtained from seeds of the tropical plant Raphia taedigera. Co-cultivation of this fungus either with Bacillus subtilis or with Streptomyces lividans resulted in the production of two new o-aminobenzoic acid derivatives, bionectriamines A and B (7 and 8) as well as of two additional known compounds (9 and 10). None of the latter compounds (7-10) were detected in axenic cultures of the fungus or of the bacteria indicating activation of silent biogenetic gene clusters through co-cultivation with bacteria. The structures of the new compounds were unambiguously determined based on detailed NMR and MS spectroscopic analysis and by comparison with the literature. The crystal structure of agathic acid (6) is reported here for the first time. Penicolinate A (4) exhibited potent cytotoxic activity against the human ovarian cancer cell line A2780 with an IC50 value of 4.1µM.
Assuntos
Alcaloides/isolamento & purificação , Arecaceae/microbiologia , Técnicas de Cocultura , Hypocreales/química , Antineoplásicos/isolamento & purificação , Bacillus subtilis , Linhagem Celular Tumoral , Endófitos/química , Humanos , Estrutura Molecular , Piridinas/isolamento & purificação , Pirróis/isolamento & purificação , Sementes/microbiologia , Streptomyces lividans , ortoaminobenzoatos/isolamento & purificaçãoRESUMO
Extracts from an endophytic fungus isolated from the roots of the Algerian plant Globularia alypum showed prominent antimicrobial activity in a screening for novel antibiotics. The producer organism was identified as Dendrothyrium variisporum by means of morphological studies and molecular phylogenetic methods. Studies on the secondary metabolite production of this strain in various culture media revealed that the major components from shake flasks were massarilactones D (1) and H (2) as well as two new furanone derivatives for which we propose the trivial names (5S)-cis-gregatin B (3) and graminin D (4). Scale-up of the fermentation in a 10 L bioreactor yielded massarilactone D and several further metabolites. Among those were three new anthranilic acid derivatives (5-7), two known anthranilic acid analogues (8 and 9) and three cyclopeptides (10-12). Their structures were elucidated on the basis of extensive spectroscopic analysis (1D- and 2D-NMR), high-resolution mass spectrometry (HRESIMS), and the application of the modified Mosher's method. The isolated metabolites were tested for antimicrobial and cytotoxic activities against various bacteria, fungi, and two mammalian cell lines. The new Metabolite 5 and Compound 9 exhibited antimicrobial activity while Compound 9 showed cytotoxicity activity against KB3.1 cells.
Assuntos
Anti-Infecciosos/química , Ascomicetos/metabolismo , Furanos/química , ortoaminobenzoatos/química , Animais , Antibacterianos/química , Antibacterianos/isolamento & purificação , Antibacterianos/farmacologia , Anti-Infecciosos/isolamento & purificação , Antifúngicos/química , Antifúngicos/isolamento & purificação , Antifúngicos/farmacologia , Antineoplásicos/química , Antineoplásicos/isolamento & purificação , Antineoplásicos/farmacologia , Reatores Biológicos , Extratos Celulares/química , Linhagem Celular , Sobrevivência Celular , Fermentação , Furanos/isolamento & purificação , Furanos/farmacologia , Humanos , Filogenia , Metabolismo Secundário , Relação Estrutura-Atividade , ortoaminobenzoatos/isolamento & purificação , ortoaminobenzoatos/farmacologiaRESUMO
Five previously undescribed metabolites, including acetylquestinol, two prenylated indole 3-carbaldehyde derivatives, an anthranilic acid derivative and an isochromone derivative, were isolated, in addition to eleven known compounds: palmitic acid, ergosterol 5,8-endoperoxide, emodin, physcion, questin, questinol, (11S, 14R)-cyclo(tryptophylvalyl), preechinulin, neoechinulin E, echinulin and eurocristatine, from the culture of the endophytic fungus Eurotium chevalieri KUFA 0006. The structures of the previously undescribed compounds were established based on an extensive 1D and 2D NMR spectral analysis as well as HRMS and IR data. In case of 2-(2, 2-dimethylcyclopropyl)-1H-indole-3-carbaldehyde and 6, 8-dihydroxy-3-(2S-hydroxypropyl)-7-methylisochromone, the absolute configurations of their stereogenic carbons were established based on comparison of their experimental and calculated ECD spectra. All the compounds, except for palmitic acid and ergosterol 5, 8-endoperoxide, were evaluated for their antibacterial and antibiofilm activities against two Gram-positive and two Gram-negative bacteria, as well as multidrug-resistant isolates from the environment. Emodin not only exhibited moderate antibacterial activity against the Gram-positive bacteria but also showed strong synergistic association with oxacillin against MRSA Staphylococcus aureus.
Assuntos
Antibacterianos/química , Biofilmes/efeitos dos fármacos , Eurotium/química , Antraquinonas/química , Antraquinonas/isolamento & purificação , Antibacterianos/isolamento & purificação , Cromonas/química , Cromonas/isolamento & purificação , Emodina/química , Emodina/isolamento & purificação , Indóis/química , Indóis/isolamento & purificação , Staphylococcus aureus Resistente à Meticilina/efeitos dos fármacos , Testes de Sensibilidade Microbiana , Estrutura Molecular , Rhizophoraceae/microbiologia , ortoaminobenzoatos/química , ortoaminobenzoatos/isolamento & purificaçãoRESUMO
A bioassay-guided investigation in conjunction with chemical screening led to the isolation of three new glycosides, ulleungoside (1), 2-methylaminobenzoyl 6-deoxy-α-l-talopyranoside (2), and naphthomycinoside (3), along with three known secondary metabolites (5-7) from Streptomyces sp. KCB13F030. Their structures were elucidated by detailed NMR and MS spectroscopic analyses. Absolute configurational analysis of the sugar units based on the magnitudes of the coupling constants, NOESY correlations, chemical derivatization, and optical rotation measurements revealed that compounds 1-3 and 5 incorporate the rare deoxyhexose 6-deoxy-α-l-talopyranose. The absolute configuration of a polyketide extender unit of 3 was determined by applying the J-based configuration analysis and modified Mosher's method. Ulleungoside (1) and naphthomycin A (7) showed in vitro inhibitory effects against indoleamine 2,3-dioxygenase activity. Further bioevaluation revealed that compounds 1 and 7 had moderate antiproliferative activities against several cancer cell lines, and compounds 5 and 6, which are members of the piericidin family, induced autophagosome accumulation.
Assuntos
Glicosídeos/química , Glicosídeos/isolamento & purificação , Glicosídeos/farmacologia , Indolamina-Pirrol 2,3,-Dioxigenase/antagonistas & inibidores , Indolamina-Pirrol 2,3,-Dioxigenase/química , Indolamina-Pirrol 2,3,-Dioxigenase/isolamento & purificação , Naftoquinonas/química , Naftoquinonas/isolamento & purificação , Policetídeos/química , Streptomyces/química , ortoaminobenzoatos/química , Bioensaio , Ensaios de Seleção de Medicamentos Antitumorais , Humanos , Espectroscopia de Ressonância Magnética , Estrutura Molecular , Naftoquinonas/farmacologia , Ressonância Magnética Nuclear Biomolecular , Policetídeos/isolamento & purificação , Policetídeos/farmacologia , ortoaminobenzoatos/isolamento & purificação , ortoaminobenzoatos/farmacologiaRESUMO
Recently, the investigation of bioactive constituents from natural products has been performed extensively around the world, yet many plants remain to be studied, even now. The author is especially interested in saponins, which are included in various natural products and Kampo medicines. Based on these study aims, we isolated the chemical constituents of Dianthus japonicus and Clethra barbinervis. The aerial parts of Dianthus japonicus were isolated into nine new (1-9) and seven known oleanane-type triterpene saponins. The structural elucidation of their chemical constituents was examined by several spectroscopic methods. Two new C-glycosyl flavones, two glycosidic derivative of anthranilic acid amides and a maltol glucoside were also isolated from this plant. The isolated compounds were evaluated according to cytotoxic inhibition toward A549 cell lines. Compounds 7-9 showed moderate inhibitory activity. The isolation of a MeOH extract of Clethra barbinervis leaves led to three new triterpene glucosides, i.e. an ursane (ryobunin A, 22), a seco-ursane (ryobunin B, 23) and an oleanane-type glucoside (ryobunin C, 24), along with four known compounds.
Assuntos
Clethraceae/química , Dianthus/química , Saponinas/isolamento & purificação , Células A549 , Antibacterianos , Antineoplásicos Fitogênicos , Produtos Biológicos , Relação Dose-Resposta a Droga , Flavonas/química , Flavonas/isolamento & purificação , Flavonas/farmacologia , Glucosídeos/química , Glucosídeos/isolamento & purificação , Glucosídeos/farmacologia , Humanos , Japão , Saponinas/química , Saponinas/farmacologia , Triterpenos/isolamento & purificação , ortoaminobenzoatos/química , ortoaminobenzoatos/isolamento & purificação , ortoaminobenzoatos/farmacologiaRESUMO
Extensive isolation work on the 1-BuOH-soluble fraction of a MeOH extract of the aerial parts of Dianthus japonicus afforded three further triterpene glycosyl estsers, termed dianthosaponins G-I, an anthranilic acid amide glucoside and a C-glycosyl flavonoid along with one known triterpene saponin. Their structures were elucidated from spectroscopic evidence. The cytotoxicity of the isolated compounds toward A549 cells was evaluated.
Assuntos
Antineoplásicos Fitogênicos/uso terapêutico , Dianthus/química , Neoplasias Pulmonares/tratamento farmacológico , Fitoterapia , Saponinas/farmacologia , Triterpenos/farmacologia , Células A549 , Adenocarcinoma Bronquioloalveolar/tratamento farmacológico , Amidas/química , Antineoplásicos Fitogênicos/química , Antineoplásicos Fitogênicos/isolamento & purificação , Antineoplásicos Fitogênicos/farmacologia , Flavonoides/química , Flavonoides/isolamento & purificação , Flavonoides/farmacologia , Glucosídeos/química , Glucosídeos/isolamento & purificação , Glucosídeos/farmacologia , Glicosídeos/química , Glicosídeos/isolamento & purificação , Glicosídeos/farmacologia , Humanos , Componentes Aéreos da Planta/química , Extratos Vegetais/química , Extratos Vegetais/farmacologia , Extratos Vegetais/uso terapêutico , Saponinas/química , Saponinas/isolamento & purificação , Saponinas/uso terapêutico , Triterpenos/química , Triterpenos/isolamento & purificação , Triterpenos/uso terapêutico , ortoaminobenzoatos/química , ortoaminobenzoatos/isolamento & purificação , ortoaminobenzoatos/farmacologiaRESUMO
CONTEXT: More analgesic compounds are needed in medicine against pain since the available drugs displayed secondary effects. Natural products are a source of molecules to develop new analgesics, using the information of plants, applied against pain, with effects such as pungency, tingling, and needle, due to their possible role in the central nervous system (NCS). Citrus reticulata Blanco (Rutaceae) leaves are usually bitten to flavor the mouth and possess this type effect in lips and tongues; due to this fact the structure of the bioctive compound could be the source of other types of analgesics. OBJECTIVE: The objective of this study is to determine the causal agent of the pungent effect in mandarin essential oil. MATERIALS AND METHODS: Mandarin essential oil was obtained and then purified by column chromatography. Each fraction was tested and pungency was detected only in the first fraction which was pure. RESULTS: The compound responsible for the pungency in the essential oils of leaves from Citrus reticulata (mandarin) was purified and the structure was assigned as methyl-N-methylanthranilate, on the basis of NMR 1D and 2D and MS. This substance corresponds to another type of molecule involving an antinociceptive effect. CONCLUSIONS: Terpenes are compounds found in essential oils. The compound responsible for the pungency of mandarin and other citrus leaves was isolated, and surprisingly it was identified as a methyl-N-methylanthranilate. This kind of molecules with this activity could be used to discover new analgesics in human therapy against pain.
Assuntos
Citrus , Óleos Voláteis/química , Óleos Voláteis/isolamento & purificação , Folhas de Planta/química , ortoaminobenzoatos/química , ortoaminobenzoatos/isolamento & purificaçãoRESUMO
Saccharomyces cerevisiae has been proven to be a valuable tool for the expression of plant metabolic pathways. By engineering a S. cerevisiae strain with two plant genes (4cl-2 from tobacco and hct from globe artichoke) we previously set up a system for the production of two novel phenolic compounds, N-(E)-p-coumaroyl-3-hydroxyanthranilic acid (Yeast avenanthramide I, Yav I) and N-(E)-caffeoyl-3-hydroxyanthranilic acid (Yeast avenanthramide II, Yav II). These compounds have a structural similarity with a class of bioactive oat compounds called avenanthramides. By developing a fermentation process for the engineered S. cerevisiae strain, we obtained a high-yield production of Yav I and Yav II. To examine the biological relevance of these compounds, we tested their potential antioxidant and antiproliferative properties upon treatment of widely used cell models, including immortalized mouse embryonic fibroblast cell lines and HeLa cancer cells. The outcomes of our experiments showed that both Yav I and Yav II enter the cell and trigger a significant up-regulation of master regulators of cell antioxidant responses, including the major antioxidant protein SOD2 and its transcriptional regulator FoxO1 as well as the down-regulation of Cyclin D1. Intriguingly, these effects were also demonstrated in cellular models of the human genetic disease Cerebral Cavernous Malformation, suggesting that the novel phenolic compounds Yav I and Yav II are endowed with bioactive properties relevant to biomedical applications. Taken together, our data demonstrate the feasibility of biotechnological production of yeast avenanthramides and underline a biologically relevant antioxidant activity of these molecules.
Assuntos
Antineoplásicos Fitogênicos/farmacologia , Antioxidantes/farmacologia , Genes de Plantas , Espécies Reativas de Oxigênio/antagonistas & inibidores , Saccharomyces cerevisiae/genética , ortoaminobenzoatos/farmacologia , Animais , Antineoplásicos Fitogênicos/biossíntese , Antineoplásicos Fitogênicos/isolamento & purificação , Antioxidantes/isolamento & purificação , Antioxidantes/metabolismo , Transporte Biológico , Linhagem Celular Transformada , Ciclina D1/antagonistas & inibidores , Ciclina D1/genética , Ciclina D1/metabolismo , Cynara scolymus/química , Cynara scolymus/genética , Fibroblastos/citologia , Fibroblastos/efeitos dos fármacos , Fibroblastos/metabolismo , Proteína Forkhead Box O1 , Fatores de Transcrição Forkhead/agonistas , Fatores de Transcrição Forkhead/genética , Fatores de Transcrição Forkhead/metabolismo , Regulação da Expressão Gênica , Células HeLa , Hemangioma Cavernoso do Sistema Nervoso Central/tratamento farmacológico , Hemangioma Cavernoso do Sistema Nervoso Central/genética , Hemangioma Cavernoso do Sistema Nervoso Central/metabolismo , Humanos , Engenharia Metabólica , Camundongos , Modelos Biológicos , Espécies Reativas de Oxigênio/metabolismo , Saccharomyces cerevisiae/metabolismo , Transdução de Sinais , Superóxido Dismutase/genética , Superóxido Dismutase/metabolismo , Nicotiana/química , Nicotiana/genética , Transgenes , ortoaminobenzoatos/isolamento & purificação , ortoaminobenzoatos/metabolismoRESUMO
Avenanthramides are characteristic constituents of oat seeds. We analyzed the methanol extract of oat seeds by HPLC and detected three compounds 1, 2, and 3 eluted at retention times similar to avenanthramides. The three compounds were purified by column chromatography and HPLC. Spectroscopic analyses of 1, 2, and 3 suggested that they are amides of 4,5-dihydroxyanthranilic acid with caffeic, p-coumaric, and ferulic acids, respectively. Their identities were confirmed by comparing spectra and chromatographic behavior with compounds synthesized from 4,5-dihydroxyanthranilic acid and N-hyrdroxysuccinimide esters of hydroxycinnamic acids. LC-MS/MS analysis with multiple reaction monitoring showed that the amounts of 1, 2, and 3 were 16.5-26.9% of corresponding avenanthamides with 5-hydroxyanthranilic acid. Compounds 1, 2, and 3 showed stronger 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical-scavenging activity than the corresponding avenanthramides with 5-hydroxyanthranilic acid, indicating the involvement of 4,5-dihydroxyanthranilic acid moiety in the scavenging of DPPH radicals.