Transcriptional analysis of Rhazya stricta in response to jasmonic acid

BACKGROUND: Jasmonic acid (JA) is a signal transducer molecule that plays an important role in plant development and stress response; it can also efficiently stimulate secondary metabolism in plant cells. RESULTS: RNA-Seq technology was applied to identify differentially expressed genes and study the time course of gene expression in Rhazya stricta in response to JA. Of more than 288 million total reads, approximately 27% were mapped to genes in the reference genome. Genes involved during the secondary metabolite pathways were up- or downregulated when treated with JA in R. stricta. Functional annotation and pathway analysis of all up- and downregulated genes identified many biological processes and molecular functions. Jasmonic acid biosynthetic, cell wall organization, and chlorophyll metabolic processes were upregulated at days 2, 6, and 12, respectively. Similarly, the molecular functions of calcium-transporting ATPase activity, ADP binding, and protein kinase activity were also upregulated at days 2, 6, and 12, respectively. Time-dependent transcriptional gene expression analysis showed that JA can induce signaling in the phenylpropanoid and aromatic acid pathways. These pathways are responsible for the production of secondary metabolites, which are essential for the development and environmental defense mechanism of R. stricta during stress conditions. CONCLUSIONS: Our results suggested that genes involved in flavonoid biosynthesis and aromatic acid synthesis pathways were upregulated during JA stress. However, monoterpenoid indole alkaloid (MIA) was unaffected by JA treatment. Hence, we can postulate that JA plays an important role in R. stricta during plant development and environmental stress conditions.

A new alkaloid from Ervatamia hainanensis / 中国中药杂志

In order to study the alkaloids from branches and leaves of Ervatamia hainanensis, silica gel, ODS, Sephadex LH-20 and HPLC chromatography were used to obtain six alkaloids from the branches and leaves of E. hainanensis with use of. Based on the physicochemical properties and spectral data, their structures were identified as 10-hydroxydemethylhirsuteine(1), 3R-hydroxycoronaridine(2), 3-(2-oxopropyl)coronaridine(3), pandine(4), 16-epi-vobasine(5), and 16-epi-vobasinic acid(6). Among them, compound 1 was a new monoterpenoid indole alkaloid, and compounds 5 and 6 were obtained from this plant for the first time.

Chemical constituents from pericarps of Aquilaria yunnanensis / 中草药

Objective: To investigate the chemical constituents from the pericarps of Aquilaria yunnanensis. Methods: The chemical constituents were separated and purified by silica gel, Sephadex LH-20 column chromatography, and semi-preparative HPLC. The structures of isolated compounds were identified by physicochemical properties and spectroscopic data. Results: Thirteen compounds were isolated from the ethyl acetate layer of 95% EtOH extract of the pericarps of A. yunnanensis, and identified as trans-linalool-3,6-oxide-7-O-β-D-(6'-O-acetyl)-glucoside (1), phenethyl-8-O-β-D-(6'-O-acetyl)-glucoside (2), mangiferin (3), iriflophenone-3,5-C-β-D-diglucoside (4), kaempferol-3-O-β-D-glucoside (5), luteolin-7-O-β-D-glucoside (6), isorhamnetin-3-O-β-D- glucoside (7), kaempferol-3-O-β-D-(6″-p-coumaroyl)-β-D-glucoside (8), geraniol-1-O-β-D-glucoside (9), 3-[2-formyl-5- (hydroxymethyl)-1H-pyrrol-1-yl] pentanedioic acid (10), cannabisin D (11), icariside D2 (12), and coniferin (13). Conclusion: Compound 1 is a new compound and compound 2 is a new natural product. Compounds 7, 9-13 were obtained from the Aquilaria genus for the first time. All compounds were firstly isolated from A. yunnanensis.

Anthelmintic activity of Eucalyptus citriodora essential oil and its major component, citronellal, on sheep gastrointestinal nematodes / Atividade anti-helmíntica do óleo essencial de Eucalyptus citriodora e seu componente majoritário, citronelal, sobre nematoides gastrintestinais de ovinos

Abstract This study aimed to evaluate the anthelmintic activity of Eucalyptus citriodora essential oil and citronellal on sheep gastrointestinal nematodes. Essential oil composition was determined by gas chromatography mass spectrometry. The substances were evaluated in vitro using adult worm motility test (AWMT) and transmission electron microscopy (TEM). The acute toxicity test in mice and the fecal egg count reduction test (FECRT) in sheep were performed. Citronellal was confirmed as the essential oil major constituent (63.9%). According to the AWMT, 2 mg/mL of essential oil and citronellal completely inhibited Haemonchus contortus motility at 6 h post exposure. H. contortus exposed to essential oil and citronellal exhibited internal ultrastructural modifications. The lethal dose 50 values in mice were 5,000 and 2,609 mg/kg for essential oil and citronellal, respectively. E. citriodora essential oil reduced sheep epg at 14 days post treatment by 69.5% (P<0.05). No significant differences were observed in epg between the citronellal and negative control groups (P>0.05). The interaction between citronellal and other constituents in the essential oil may be relevant for its in vivo anthelmintic activity. Thus, E. citriodora essential oil and citronellal pharmacokinetic studies may help elucidate the anthelmintic activity of these compounds.

Resumo Este trabalho objetivou avaliar a atividade anti-helmíntica do óleo essencial de Eucalyptus citriodora e citronelal sobre nematoides gastrintestinais de ovinos. A composição do óleo essencial foi determinada por cromatografia gasosa acoplada à espectrometria de massas. As substâncias foram avaliadas in vitro utilizando-se teste de motilidade de vermes adultos (AWMT) e microscopia eletrônica de transmissão (TEM). Teste de toxicidade aguda em camundongos e teste de redução da contagem de ovos fecais (FECRT) em ovinos foram realizados. Citronelal foi confirmado como componente majoritário do óleo essencial (63,9%). No AWMT, 2 mg/mL de óleo essencial e citronelal inibiram completamente a motilidade de H. contortus 6 h pós-exposição. H. contortus expostos ao óleo essencial e citronelal exibiram modificações ultraestruturais internas. Os valores da dose letal 50 em camundongos foram 5.000 e 2.609 mg/kg para óleo essencial e citronelal, respectivamente. Óleo essencial de E. citriodora reduziu opg de ovinos 14 dias pós-tratamento em 69,5% (P<0,05). Não houve diferença significativa de opg entre grupo controle negativo e citronelal (P>0,05). A interação entre citronelal e outros constituintes do óleo essencial pode ser relevante na atividade anti-helmíntica in vivo. Portanto, avaliação farmacocinética do óleo essencial de E. citriodora e citronelal pode auxiliar a elucidar a atividade anti-helmíntica desses compostos.

Research progress on pharmacological properties and monoterpenoid indole alkaloids biosynthetic pathway in plants of genus Rauvolfia / 中草药

Rauvolfia serpentine is a traditional Indian medicine and now the genus Rauvolfia becomes a commonly used south medicine in China. Rauvolfia is mainly used to treat hypertension, mental illness, cardiovascular disorders, cancer, and other diseases. The paper overviewed the main species and distribution, pharmacological properties, and the monoterpenoid indole alkaloids biosynthetic pathway of Rauvolfia in China. The paper aims to provide reference for research and clinical application in plants of the genus Rauvolfia.

A new monoterpenoid glycoside from aerial parts of Siegesbeckia pubescens / 中草药

Objective: To study the chemical constituents of the aerial parts of Siegesbeckia pubescens. Methods: A systematic separation of chemical constituents was conducted. Detailed chemical investigation of S. pubescens led to the isolation of six compounds by comprehensive chromatographic Methods: (Extraction, Silic gel C.C, Sephadex LH-20, ODS C. C). The structures of them were fully determined based on spectroscopic analysis including IR, HR-ESI-MS, ESI-MS, 1H-NMR, 13C-NMR, DEPT, HSQC, and HMBC spectrum. Results: A total of six compounds were isolated from the fraction of n-butanol extract of S. pubescens including one new monoterpenoid glycoside named (2Z,5E)-7-hydroxy-3,7-dimethyl-2,5-octadiene-1-O-[α-L-rhamnpyranosyl (1→6)]-β-D-glucopyranoside (1), four known diterpenoid glucosides named pubeside D (2), ent-15,16,19-trihydroxypimar- 8(14)-en-2-one-19-O-β-glucopyranoside (3), ent-15,16-dihydroxypimar-8(14)-en-2-one-19-oic-β-glucopyranoside (4), and neodarutoside (5), and eleutherazine B (6). Conclusion: Compound 1 is a new compound, named as siegeside F, and compound 6 is isolated from this species for the first time.

Study of the antifungal potential of (r)-(+)-citronellal and its association with therapeutic agents used in the treatment of vulvovaginal candidiasis / Estudo do potencial antifúngico do (r)-(+)-citronelal e sua associação com agentes terapêuticos utilizados no tratamento da candidíase vulvovaginal

Vulvovaginal candidiasis (VVC) is a common fungal infection that affects healthy women of all ages. At least 75% of women will develop one or more infections once during their lifetime, with 6 to 9% of those individuals developing recurrent infections. In view of this context, this study sought to evaluate the antifungal potential of the isolated (R)-(+)-citronellal [(R)-(+)-CT] and associated to therapeutic agents of clinical importance. The enantiomer was solubilized in tween 80 and dimethylsulfoxide (DMSO). Posteriorly diluted in sterile distilled water up to the concentration of 2048µg/mL. The minimum inhibitory concentration (MIC) of the product was determined by microdilution in RPMI-1640 obtaining dilutions of 1024-4µg/mL. The minimum fungicidal concentration (MFC) was determined by the Sabouraud dextrose agar (SDA) depletion technique from aliquots of 1µL of the MIC, MIC × 2 and MIC × 4. The MIC and the MFC values of (R)-(+)-CT for 90% of the C. albicans strains were 16 and 32µg/mL respectively. In the susceptibility test, C. albicans presented a high resistance to fluconazole and to itraconazole, 12 (92.30%) of the strains. However, for ketoconazole and miconazole the resistance was of 4 (30.76%) and 3 (23.07%) of the strains respectively. In the combination testing of the (R)-(+)-CT with ketoconazole and miconazole, the resistance was completely reverted. For fluconazole and itraconazole, the resistance was reverted in 9 (75%) and 7 (58.33%) of the strains respectively. The (R)-(+)-CT presented fungicide activity with MFC of MIC × 2. When in combination with ketoconazole, fluconazole, itraconazole and miconazole increased the inhibition zones of these antifungal drugs, reducing the resistance against C. albicans.

Candidíase vulvovaginal (CVV) é uma infecção fúngica comum que afeta mulheres saudáveis de todas as idades. Pelo menos 75% das mulheres irão desenvolver uma ou mais infecções uma vez durante a vida, com 6 a 9% dos indivíduos desenvolvendo infecções recorrentes. Diante deste contexto, buscou-se avaliar neste estudo o potencial antifúngico do (R)-(+)-citronelal [(R)-(+)-CT] isolado e associado a agentes terapêuticos de importância clínica. O enantiômero foi solubilizado em tween 80 e dimetilsulfóxido (DMSO). Posteriormente diluiu-se em água destilada estéril até a concentração de 2048µg/mL. A concentração inibitória mínima (CIM) do produto foi determinada por microdiluição em meio RPMI-1640 obtendo diluições de 4-1024µg/mL. A concentração fungicida mínima (CFM) foi determinada pela técnica de esgotamento em agar Sabouraud dextrose (ASD) a partir de alíquotas de 1mL da CIM, CIM × 2 e CIM × 4. A CIM e a CFM do (R)-(+)-CT para 90% das cepas de C. albicans foram 16 e 32µg/mL respectivamente. No ensaio de suscetibilidade, C. albicans apresentou alta resistência ao fluconazol e ao itraconazol, 12 (92.30%) das cepas. No em tanto, para o cetoconazol e o miconazol a resistência foi de 4 (30.76%) e 3 (23.07%) das cepas respectivamente. No ensaio de combinação do (R)-(+)-CT com cetoconazol e miconazol, a resistência foi completamente revertida. Para o fluconazol e o itraconazol, a resistências foi revertida em 9 (75%) e 7 (58.33%) das cepas respectivamente. O (R)-(+)-CT apresentou atividade fungicida com CFM igual à CIM × 2. Quando em combinação com cetoconazol, fluconazol, itraconazol e miconazol ampliou as zonas de inibição desses antifúngicos, diminuindo a resistência contra C. albicans.

Study on alkaloids from Ervatamia hainanensis / 中草药

Objective: To study the alkaloids from Ervatamia hainanensis. Methods: The alkaloids were isolated and purified by silica gel, MCI, Sephadex LH-20 column chromatography, and semi-preparative HPLC, and their structures were elucidated by physical and spectroscopic analysis. Results: Twelve alkaloids were obtained and identified as coronaridine (1), 19-epi-heyneanine (2), 9,10-dimethoxycoronaridine (3), vobasine (4), vobasine N(4)-oxide (5), 3-oxo-19-epi-heyneanine (6), strictamine (7), deacetylakuammiline (8), pandine (9), rhazicine (10), rhazicine N(4)-oxide (11), and rhazimine (12). Conclusion: Compounds 8 and 10-12 are isolated from the genus Ervatamia Stapf for the first time, while compounds 3 and 5-7 are firstly obtained from E. hainanensis.

Chemical constituents from stems of Yao medicine Zhongliuteng (cane of Pileostegia tomentella) / 中草药

Objective: To study the chemical constituents of Yao medicine Zhongliuteng, the canes of Pileostegia tomentella. Methods: The chemical constituents were isolated and purified by silica gel chromatography repeatedly from the canes of P. tomentella and their structures were identified by spectral analysis and chemical methods. Results: Sixteen compounds were isolated from the canes of P. tomentella and their structures were identified as loganic acid (1), 4-O-α-L-arabinofuranosyl-(1→6)-β-D-glucopyranoside (4S)-4-hydroxyl-β-ionone (2), resorcinol (3), daphnin (4), skimmin (5), umbelliferone (6), vogeloside (7), diethyl phthalate (8), secologanin (9), secologanin dimethyl acetal (10), sweroside (11), foliasalacioside B (12), benzyl-O-β-D-apiofuranosyl-(1″→6')-β-D-glucopyranoside (13), maltose (14), glucose (15), and daucosterol (16). Conclusion: All compounds are obtained from the plant for the first time, and all compounds are also obtained from the plants of Pileostegia Hook. f. et Thoms. for the first time except compounds 5, 6 and 16.

A new monoterpenoid indole alkaloid from Gardneria multiflora Makino / 中国药学杂志

OBJECTIVE: To study the monoterpenoid indole alkaloids in Gardneria multiflora Makino. METHODS: The dried plants of G. multiflora Makino were extracted with 95% ethanol. After removal of solvent, the residue was diluted with water, then extracted with petroleum ether and chloroform to obtain crude alkaloids. Column chromatograghy on silica gel, Rp-18, and Sephadex LH-20 were applied for the isolation and purification of crude alkaloids. The structure was elucidated based on detailed spectroscopic evidence. RESULTS: One monoterpenoid indole alkaloid was isolated from this plant. CONCLUSION: Compound gardmultimine A (1) is a new compound.

A new monoterpenoid indole alkaloid from Winchia calophylla / 中草药

To investigate the alkaloid constituents from the trunk barks of Winchia calophylla. The constituents were isolated and purified by column chromatography over silica gel and Sephadex LH-20 columns. The structure of the isolated compound was elucidated by MS and NMR spectra, and then confirmed by a single-crystal X-ray crystallographic analysis. The isolated compound was also evaluated for its in vitro cytotoxicity against the proliferation of seven kinds of human cancer cell lines (lung cancer: A549, breast cancer: MCF-7, prostate cancer: PC-3, glioma: U87MG, multiple myeloma: U266, MM1. S, and MM1. R). A new monoterpenoid indole alkaloid, identified as calophyline B (1), was isolated from the ethanol extract of the trunk barks of W. calophylla. Compound 1 is a new compound, which shows no cytotoxicity against the proliferation of the above human tumor cells.

Water soluble chemical constituents of Valeriana officinalis Linn. var. latiofolia Miq / 第二军医大学学报

Objective To investigate the water soluble chemical constituents in the root of Valeriana officinalis Linn. var. latio folia Miq. and to identify their structures. Methods The water soluble chemical constituents were isolated and purified by macroporous resin Diaion HP-20, silica gel and Sephadex LH-20; the chemical structures of the components were determined by MS, 1D NMRand 2D NMR spectral analysis. Results and conclusion Thirteen constituents were isolated and identified from the root o f Valeriana officinalis Linn. var. latio folia, including Ganxinoside A (1), (-)-angelicoideno 1-2-0-β-D-glucopyranoside (2), (-)-angelicoidenol-2-0-β-D-apiofuranosyl-(1→6)-β-D-glucopyranoside (3), 6-hydroxy-7-(hydroxymethyl)-4-methylenehexahydrocyclopenta [c] pyran-1 (3H)-one (4), kanokoside A (5), prinsepiol-4-O-β-D-glucopyranoside (6), (+)-pinoresinol-4-0-β-D-glucopyranoside (7), sinenoside I (8), lanicepsides A (9), (+)-1-hydroxypinoresinol-1-O-β-D-glucoside (10), (+)-pinoresinol-4, 4′-0-bisglucopyranoside (11), (+)-cycloolivil 6-O-β-D-glucopyranoside (12), and coniferin (13). All the 13 compounds are isolated from the root o f Valeriana officinalis Linn. var. latio folia Miq. for the first time.

Chemical constituents from rhizomes of Paeonia sinjiangensis / 中草药

Objective: To investigate the chemical constituents from the rhizomes of Paeonia sinjiangensis. Methods: The chemical constituents were isolated and identified by chromatography on silica gel, Sephadex LH-20, ODS, and RP-HPLC. Their structures were elucidated on the basis of physicochemical properties and spectral analyses. Results: Sixteen compounds were isolated from the 80% ethanol extract from the rhizomes of P. sinjiangensis, and identified as 8-debenzoylpaeoniflorin (1), 4-O-butylpaeoniflorin (2), galloylpaeoniflorin (3), 4-O-methylbenzoylpaeoniflorin (4), benzoylalbiflorin (5), albiflorin R1 (6), 9-O-butylpaeonidanin (7), (Z)-(1S, 5R)-β-pinen-10-yl β-vicianoside (8), gallic acid (9), methyl gallate (10), ethyl gallate (11), isovanillic (12), 6-hydroxycoumarin (13), 3, 4, 23-trihydroxy-24, 30-dinorolean-12, 20 (29)-dien-28-oic acid (14), palbinone (15), and cadina-4, 11-dien-14-oic acid (16). Conclusion: Compound 16 is firstly isolated from the Ranunculaceae family, and compounds 1-15 are isolated from this plant for the first time.

Amine-lyases and Their Applications in Preparation of Pharmaceutical Intermediates / 微生物学通报

Carbon-nitrogen lyases (E.C.4.3) are a group of enzymes that release ammonia, amidine or amino group etc, and also show ability to form double bond or ring structure. Specifically, enzymes forming amino group are called amine-lyases (E.C.4.3.3), which are critical in the industrial production of many medicine intermediates. In this review is a summary of four major amine-lyases in terms of their source, enzymatic characteristics and their applications in preparation of pharmaceutical intermediates.