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This study established an Orthogonal Partial Least Squares (OPLS) model combining 1H-NMR and GC-MS data to identify characteristic metabolites in complex extracts. Both in metabolomics studies, and natural product chemistry, the reliable identification of marker metabolites usually requires laborious isolation and purification steps, which remains a bottleneck in many studies. Both ginger (GR) and processed ginger (PGR) are listed in the Japanese pharmacopeia. The plant of origin, the rhizome of Zingiber officinale Roscoe, is differently processed for these crude drugs. Notably, the quality of crude drugs is affected by genetic and environmental factors, making it difficult to maintain a certain quality standard. Therefore, characteristic markers for the quality control of GR and PGR are required. Metabolomic analysis using 1H-NMR was able to discriminate between GR and PGR, but there were unidentified signals that were difficult to distinguish based on NMR data alone. Therefore, we combined 1H-NMR and GC-MS analytical data to identify them by OPLS. As a result, αr-curcumene was found to be a useful marker for these identifications. This new approach enabled rapid identification of characteristic marker compounds and reduced the labor involved in the isolation process.
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Methylmercury (MeHg) is widely distributed in nature and is known to cause neurotoxic effects. This study aimed to examine the anti-MeHg activity of oleanolic acid-3-glucoside (OA3Glu), a synthetic oleanane-type saponin derivative, by evaluating its effects on motor function, pathology, and electrophysiological properties in a mouse model of MeHg poisoning. Mice were orally administered 2 or 4â¯mg·kg-1·d-1 MeHg with or without 100⯵g·kg-1·d-1 OA3Glu 5x/week for four weeks. Motor function was evaluated using beam-walking and dynamic weight-bearing (DWB) tests. High-dose MeHg exposure significantly increased the frequency of stepping off the hind leg while crossing the beam in the beam-walking test, and increased weight on forelegs when moving freely in the DWB test. OA3Glu treatment alleviated motor abnormality caused by high-dose MeHg exposure in both motor function tests. Additionally, OA3Glu treatment reduced the number of contracted Purkinje cells frequently observed in the cerebellum of MeHg-treated groups, although cerebrum histology was similar in all experimental groups. The synaptic potential amplitude in the cerebellum decreased as MeHg exposure increased, which was restored by OA3Glu treatment. Even in the cerebrum, where the effects of MeHg were not observed, the amplitude of the field potential was suppressed with increasing MeHg exposure but was restored with OA3Glu treatment. Taken together, the study findings suggest that OA3Glu improves neurotransmission and movement disorders associated with MeHg exposure via protection of Purkinje cells in the cerebellum while ameliorating pre/post-synaptic deficits in the cerebral cortex in which no changes were observed at the tissue level, potentially providing a treatment to mitigate MeHg toxicity.
Assuntos
Compostos de Metilmercúrio , Ácido Oleanólico , Saponinas , Transmissão Sináptica , Animais , Compostos de Metilmercúrio/toxicidade , Masculino , Ácido Oleanólico/farmacologia , Ácido Oleanólico/análogos & derivados , Transmissão Sináptica/efeitos dos fármacos , Camundongos , Saponinas/farmacologia , Glucosídeos/farmacologia , Células de Purkinje/efeitos dos fármacos , Células de Purkinje/patologia , Cerebelo/efeitos dos fármacos , Cerebelo/patologia , Cerebelo/metabolismo , Atividade Motora/efeitos dos fármacos , Camundongos Endogâmicos ICRRESUMO
This study used two types of analyses and statistical calculations on powdered samples of Polygala root (PR) and Senega root (SR): (1) determination of saponin content by an independently developed quantitative analysis of tenuifolin content using a flow reactor, and (2) near-infrared spectroscopy (NIR) using crude drug powders as direct samples for metabolic profiling. Furthermore, a prediction model for tenuifolin content was developed and validated using multivariate analysis based on the results of (1) and (2). The goal of this study was to develop a rapid analytical method utilizing the saponin content and explore the possibility of quality control through a wide-area survey of crude drugs using NIR spectroscopy. Consequently, various parameters and appropriate wavelengths were examined in the regression analysis, and a model with a reasonable contribution rate and prediction accuracy was successfully developed. In this case, the wavenumber contributing to the model was consistent with that of tenuifolin, confirming that this model was based on saponin content. In this series of analyses, we have succeeded in developing a model that can quickly estimate saponin content without post-processing and have demonstrated a brief way to perform quality control of crude drugs in the clinical field and on the market.
Assuntos
Saponinas , Espectroscopia de Luz Próxima ao Infravermelho , Espectroscopia de Luz Próxima ao Infravermelho/métodos , Controle de Qualidade , Análise dos Mínimos QuadradosRESUMO
Apricot and Peach Kernels are commercial crude drugs used in many formulas of traditional Japanese medicine, Kampo. Although their applications are quite different, it is difficult to distinguish them using conventional methods such as HPLC. The study aimed at near-infrared (NIR) metabolic profiling to discriminate Apricot and Peach Kernels (Armeniacae Semen and Persicae Semen) collected from Japanese markets. A fast, simple, non-destructive, and robust NIR measurement of kernel surface with no sample pre-treatment was achieved in situ. Principal component analysis and orthogonal partial least squares discriminant analysis (OPLS-DA) models showed discrimination between the two crude drugs with good fitting and prediction values. These results indicate that NIR metabolic profiling is useful for discriminating Apricot and Peach Kernels based on their chemical constituents using a simple and non-destructive procedure.
Assuntos
Prunus armeniaca , Prunus persica , Metabolômica , Análise de Componente Principal , Cromatografia Líquida de Alta PressãoRESUMO
An improved process for preparing tenuifolin (presenegenin 3-ß-d-glucopyranoside) from the root of Polygala senega L. was developed. A crude saponin mixture extracted from P. senega was subjected to hydrolysis, and the reactivity of compounds in the extract was controlled by utilizing the combination of a flow reactor and experimental design. In addition, column chromatography with HP 20, a synthetic polystyrenic adsorbent, allowed the gram-scale preparation of tenuifolin in a continuous manner with fewer steps. This approach shortens the total time required for gram-scale preparation from 16 to 5 h in a continuous manner while improving the yield from 0.59% to 2.08% (w/w).
Assuntos
Polygala , Diterpenos do Tipo Caurano , Hidrólise , Raízes de Plantas , TemperaturaRESUMO
Triterpene and steroid saponins have various pharmacological activities but the synthesis of C-3 monodesmosidic saponins remains challenging. Herein, a series of C-3 glycosyl monodesmosidic saponins was synthesized via the microfluidic glycosylation of triterpenoids or steroids at the C-3 position, without the formation of orthoester byproducts, and subsequent deprotection of the benzoyl (Bz) group. This microfluidic glycosylation/batch deprotection sequence enabled the efficient synthesis of C-3 saponins with fewer purification steps and a shorter reaction time than conventional batch synthesis and stepwise microfluidic glycosylation. Furthermore, this system minimized the consumption of the imidate donor. Using this reaction system, 18 different C-3 saponins and 13 different C-28-benzyl-C-3 saponins, including 8 new compounds, were synthesized from various sugars and triterpenes or steroids. Our synthetic approach is expected to be suitable for further expanding the C-3 saponin library for pharmacological studies.
Assuntos
Técnicas Analíticas Microfluídicas , Saponinas/síntese química , Glicosilação , Conformação Molecular , Saponinas/químicaRESUMO
Identifying different species of the genus Atractylodes which are commonly used in Chinese and Japanese traditional medicine, using chromatographic approaches can be difficult. 1H NMR metabolic profiling of DNA-authenticated, archived rhizomes of the genus Atractylodes was performed for genetic and chemical evaluation. The ITS region of the nuclear rDNA was sequenced for five species, A. japonica, A. macrocephala, A. lancea, A. chinensis, and A. koreana. Our samples had nucleotide sequences as previously reported, except that part of the A. lancea cultivated in Japan had a type 5, hybrid DNA sequence. Principal component analysis (PCA) using 1H NMR spectra of extracts with two solvent systems (CD3OD, CDCl3) was performed. When CDCl3 extracts were utilized, the chemometric analysis enabled the identification and classification of Atractylodes species according to their composition of major sesquiterpene compounds. The 1H NMR spectra using CD3OD contained confounding sugar peaks. PCA removal of these peaks gave the same result as that obtained using CDCl3 and allowed species distinction. Such chemometric methods with multivariate analysis of NMR spectra will be useful for the discrimination of plant species, without specifying the index components and quantitative analysis on multi-components.
Assuntos
Atractylodes/química , Atractylodes/classificação , Metabolômica , Compostos Fitoquímicos/análise , Sequência de Bases , DNA de Plantas/genética , DNA Espaçador Ribossômico/genética , Japão , Espectroscopia de Ressonância Magnética , Filogenia , Análise de Componente Principal , Rizoma/química , Rizoma/genética , Sesquiterpenos/análiseRESUMO
Methylmercury (MeHg) is one of the most toxic environmental pollutants, presenting a serious health hazard worldwide. In this study, we examined the potential of derivatives of oleanolic acid (OA), such as OA 3-glucoside, OA 28-glucoside, and OA 3,28-diglucoside, to mitigate MeHg toxicity in vitro and in vivo. We found that OA 3-glucoside suppressed the cellular MeHg uptake by 63.4% compared with that of the control and improved the cell viability from 75.4% to 107.9% upon exposure to cytotoxic MeHg in Caco-2 cells. To verify the anti-MeHg activity of OA 3-glucoside, mice were orally administered MeHg (0, 1.0, or 5.0 mg kg-1·d-1), with or without OA 3-glucoside, and then mercury accumulation was measured in various organs of the mice. The mice co-treated with MeHg and OA 3-glucoside showed significantly lower mercury content in organs such as the cerebrum, cerebellum, liver, kidney, and spleen, with 83.1%, 68.7%, 71.7%, 82.1%, and 18.2% of those in the OA 3-glucoside-untreated group, respectively. This suggested OA 3-glucoside had the potential as an anti-MeHg compound, owing to its ability to suppress the distribution of MeHg into organs. Supporting this hypothesis, the mice treated with MeHg and OA 3-glucoside showed a tendency to survive one day longer than the control mice. Our findings suggest OA 3-glucoside administration alleviates the toxicity of MeHg by suppressing MeHg accumulation in organs.
Assuntos
Glucosídeos/farmacologia , Compostos de Metilmercúrio/toxicidade , Ácido Oleanólico/análogos & derivados , Ácido Oleanólico/farmacologia , Saponinas/farmacologia , Animais , Células CACO-2 , Relação Dose-Resposta a Droga , Trato Gastrointestinal/efeitos dos fármacos , Trato Gastrointestinal/metabolismo , Glucosídeos/síntese química , Humanos , Masculino , Compostos de Metilmercúrio/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Ácido Oleanólico/síntese química , Distribuição Aleatória , Saponinas/síntese químicaRESUMO
There is no drug administration-approved therapy for non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis (NASH). In this study, eight compounds, gallic acid (1), methyl gallate (2), corilagin (3), 3,4,8,9,10-pentahydroxydibenzo[b,d]pyran-6-one (4), repandinin B (5), (Z)-3-hexenyl-ß-D-glucopyranoside (6), (+)-lyoniresinol-3α-O-α-L-rhamnopyranoside (7) and mallophenol A (8) were isolated from the active fractions of Mallotus furetianus. Three compounds, (6, 7 and 8) revealed potent anti-steatosis activity in the oleic acid (OA)-induced steatosis cell model, with the minimum effective concentration of 0.05 (6), 0.0005 (7) and 0.0005 (8) µg/mL, which were much lower than the control compound, fibrate (72.4 µg/mL).
Assuntos
Mallotus (Planta)/química , Hepatopatia Gordurosa não Alcoólica/tratamento farmacológico , Folhas de Planta/química , Ácido Gálico/análogos & derivados , Ácido Gálico/isolamento & purificação , Ácido Gálico/farmacologia , Glucosídeos/isolamento & purificação , Glucosídeos/farmacologia , Células Hep G2 , Humanos , Taninos Hidrolisáveis/isolamento & purificação , Taninos Hidrolisáveis/farmacologia , Hepatopatia Gordurosa não Alcoólica/induzido quimicamente , Ácido Oleico/toxicidadeRESUMO
Described herein is a synthetic strategy for the total synthesis of (±)-phomoidrideâ D. This highly efficient and stereoselective approach provides rapid assembly of the carbocyclic core by way of a tandem phenolic oxidation/intramolecular Diels-Alder cycloaddition. A subsequent SmI2 -mediated cyclization cascade delivers an isotwistane intermediate poised for a Wharton fragmentation that unveils the requisite bicyclo[4.3.1]decene skeleton and sets the stage for synthesis completion.
Assuntos
Anidridos Maleicos/síntese química , Compostos Bicíclicos com Pontes/química , Ciclização , Reação de Cicloadição , Oxirredução , EstereoisomerismoRESUMO
A new cyclopropane-containing sesquiterpenoid, phellilane L (1), was isolated from the medicinal mushroom Phellinus linteus ("Meshimakobu" in Japanese), a member of the Hymenochaetaceae family and a well-known fungus that is widely used in East Asia. The planar structure of 1 was determined on the basis of spectroscopic analysis. The authors achieved the first total synthesis of 1. Our protecting group-free synthesis features a highly stereoselective one-pot synthesis involving an intermolecular alkylation/cyclization/lactonization strategy for construction of the key cyclopropane-γ-lactone intermediate. Additionally, our synthesis determined the absolute configuration of phellilane L (1).
Assuntos
Agaricales/química , Basidiomycota/química , Sesquiterpenos/química , Química Farmacêutica , Estrutura MolecularRESUMO
This corrects the article DOI: 10.1038/ja.2017.35.
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This corrects the article DOI: 10.1038/ja.2015.142.
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Glycyrrhetinic acid (GA) is an active metabolite of glycyrrhizin, which is a main constituent in licorice (Glycyrrhiza glabra). While GA exhibits a wide variety of pharmacological activities in the body, it is converted to a toxic metabolite GA 3-O-glucuronide by hepatic UDP-glucuronosyltransferases (UGTs). To avoid the development of the toxic metabolite-induced pseudohyperaldosteronism (pseudoaldosteronism), there is a limitation in maximum daily dosage of licorice and in combined usage of other glycyrrhizin-containing natural medicine. In this study, we investigated the inhibitory effects of various polyphenols and triterpenoids on the UGT-mediated GA 3-O-glucuronidation. In human liver microsomes, UGT-mediated GA glucuronidation was significantly inhibited by protopanaxadiol with an IC50 value of 59.2 µM. Isoliquiritigenin, rosmarinic acid, alisol B, alisol acetate, and catechin moderately inhibited the GA glucuronidation with IC50 values of 96.4 µM, 125 µM, 160 µM, 163 µM, and 164 µM. Other tested 19 polyphenols and triterpenoids, including liquiritigenin, did not inhibit UGT-mediated GA glucuronidation in human liver microsomes. Our data indicate that relatively higher dosage of licorice can be used without a risk of developing pseudohyperaldosteronism in combination of natural medicine containing protopanaxadiol such as Panax ginseng. Furthermore, supplemental protopanaxadiol and isoliquiritigenin might be useful in preventing licorice-inducing pseudoaldosteronism.
Assuntos
Inibidores Enzimáticos/farmacologia , Glucuronídeos/metabolismo , Glucuronosiltransferase/antagonistas & inibidores , Ácido Glicirretínico/metabolismo , Microssomos Hepáticos/efeitos dos fármacos , Polifenóis/farmacologia , Triterpenos/farmacologia , Relação Dose-Resposta a Droga , Glucuronosiltransferase/metabolismo , Humanos , Microssomos Hepáticos/metabolismo , Estrutura Molecular , Relação Estrutura-AtividadeRESUMO
We report the first synthesis of a series of bisdesmosidic oleanolic acid saponins using microflow reactor Comet X-01 via a continuous flow glycosylation-batch deprotection sequence. The main results of this study can be summarized as follows: (1) The microfluidic glycosylation of oleanolic acid at C-28 was achieved in quantitative yield and was applied to the synthesis of six C-28-monoglycosidic saponins. (2) The microfluidic glycosylation of oleanolic acid at C-3 was achieved in good yield without orthoester byproduct formation and was applied to the synthesis of three bisdesmosidic saponins. (3) The continuous synthesis of saponins via a microfluidic glycosylation-batch deprotection sequence was achieved in four steps involving two purifications. Thus, the continuous microfluidic glycosylation-deprotection process is expected to be suitable for the preparation of a library of bisdesmosidic oleanolic acid saponins for in vivo pharmacological studies.
Assuntos
Antibacterianos/isolamento & purificação , Extratos Vegetais/química , Porphyromonas gingivalis/efeitos dos fármacos , Sesquiterpenos/isolamento & purificação , Antibacterianos/química , Antibacterianos/farmacologia , Testes de Sensibilidade Microbiana , Phellinus , Sesquiterpenos/química , Sesquiterpenos/farmacologiaRESUMO
A series of new simplified oleanolic acid saponins with a glycosyl ester moiety at C28, were efficiently prepared. Furthermore, the effect of nasal administration of the synthetic oleanolic acid saponins on the nasal anti-influenza virus antibody titer against secondary nasal inoculation of the influenza split vaccine was examined. The result revealed cinnamoyl saponin as a suitable candidate vaccine adjuvant.
Assuntos
Adjuvantes Imunológicos/síntese química , Adjuvantes Imunológicos/farmacologia , Vacinas contra Influenza/administração & dosagem , Mucosa Nasal/efeitos dos fármacos , Ácido Oleanólico/química , Saponinas/síntese química , Saponinas/farmacologia , Adjuvantes Imunológicos/química , Administração Intranasal , Animais , Camundongos , Camundongos Endogâmicos BALB C , Saponinas/química , Análise Espectral/métodosRESUMO
A new natural product, designated iminimycin A, was isolated from the cultured broth of a streptomycin-producing microbial strain, Streptomyces griseus OS-3601, via a physicochemical screening method using HP-20, silica gel and ODS column chromatographies and subsequent preparative HPLC. Iminimycin A is an indolizidine alkaloid, containing of an unusual iminium group and a cyclopropane ring with a triene side chain. The absolute configuration of iminimycin A was elucidated by NMR studies and electronic circular dichroism analysis. Iminimycin A shows anti-bacterial activity against Bacillus subtilis, Kocuria rhizophila and Xanthomonas campestris pv. orizae, and cytotoxic activity against HeLa S3 and Jurkat cells with IC50 values of 43 and 36 µM, respectively.
Assuntos
Antibacterianos/farmacologia , Antineoplásicos/farmacologia , Streptomyces griseus/metabolismo , Alcaloides/química , Alcaloides/isolamento & purificação , Alcaloides/farmacologia , Antibacterianos/química , Antibacterianos/isolamento & purificação , Antineoplásicos/química , Antineoplásicos/isolamento & purificação , Bacillus subtilis/efeitos dos fármacos , Cromatografia Líquida de Alta Pressão/métodos , Dicroísmo Circular , Células HeLa , Humanos , Indolizidinas/química , Indolizidinas/isolamento & purificação , Indolizidinas/farmacologia , Concentração Inibidora 50 , Células Jurkat , Espectroscopia de Ressonância Magnética , Micrococcaceae/efeitos dos fármacos , Xanthomonas campestris/efeitos dos fármacosRESUMO
The total synthesis of the indole alkaloids, neoxaline, oxaline and meleagrinâ A, all containing a unique indoline spiroaminal framework, was accomplished through the stereoselective introduction of a reverse prenyl group to the congested benzylic carbon of furoindoline, a two-pot transformation of indoline (containing three nitrogen atoms at appropriate positions) to the featured indoline spiroaminal framework, and elimination of carbonate assisted by the adjacent imidazole moiety to construct the (E)-dehydrohistidine. The absolute stereochemistry of neoxaline was elucidated through our total synthesis. In addition, we evaluated the bioactivity, especially the anti-infectious properties, of neoxaline and oxaline, and of some synthetic intermediates.