Quality Analysis and Suggestion for Ginkgo Folium Tablets and Ginkgo Folium Capsules / 中国实验方剂学杂志

Objective:To conduct quality evaluation of Ginkgo Folium preparations by analyzing the national evaluation sampling test results， analyze the quality differences， and put forward suggestions for the improvement of quality standards and market supervision. Method:The contents of total flavonol glycosides and terpene lactones in Ginkgo Folium tablets and Ginkgo Folium capsules were determined according to the methods of determination in the 2015 edition of Chinese Pharmacopoeia （the first volume）， and the contents of free flavonoids （quercetin， kaempferide and isorhamnetin） and sophoricoside in Ginkgo Folium preparations were determined according to related supplementary testing method of Ginkgo Folium tablets and Ginkgo Folium capsules issued by National Medical Products Administration. The quality differences of Ginkgo Folium preparations from different batches and different manufacturers were compared according to the contents of total flavonol glycosides， terpene lactones， free flavonoids and sophoricoside in 328 batches of Ginkgo Folium tablets and Ginkgo Folium capsules manufactured by 48 enterprises. Result:Quality of 328 batches of Ginkgo Folium tablets and Ginkgo Folium capsules was in accordance with the standard， but the contents of terpene lactones and total flavonol glycosides were all distributed in a wide range， and the quality of samples varied greatly among different enterprises. Conclusion:It is recommended that each enterprise should optimize the production process and strictly control the raw materials to ensure the consistency between different batches of samples.

Sesquiterpene lactones of Aucklandia lappa: Pharmacology, pharmacokinetics, toxicity, and structure–activity relationship / 中草药·英文版

The medicinal part of Aucklandia lappa (Asteraceae) is its dried root, which is one of the commonly used Chinese medicinal materials. Here we reviewed sesquiterpene lactones isolated from A. lappa over the past ten years in the following aspects of pharmacological activities, pharmacokinetics, toxicology, structure–activity relationship. Pharmacological activities consist of anti-cancer, anti-inflammatory activity, anti-immunity activity, anti-oxidant activity, antimicrobial activity, spasmolytic activity and so on. The extractive, showing similar pharmacokinetics parameters, may exert their various biological activities by the interaction of their α-methylene-γ-butyrolactone moiety with the thiol groups of biomacromolecules through Michael-addition. However, the poor aqueous solubility, non-selective binding as a Michael acceptor at undesired targets limited clinical translation of this class. In order to evaluate the potential effect of the extractive applied in clinical trial, the present review outlines information on pharmacological activities, pharmacokinetics, toxicology, and structure–activity relationship, as well as the future research directions of the extractive for further development and utilization of A. lappa.

Germacrene-derived sesquiterpene lactones:opportunities and challenges for biosynthesis / 中国中药杂志

Sesquiterpene lactones are a kind of widely distributed natural organic compounds with anti-tumor, anti-malarial and other significant biological activities. Based on their carbocylic skeletons, sesquiterpene lactones are classified into germacranolide, guaia-nolide, xanthanolide, pseudo-guaianolide, elemonolide and eudesmanolide, etc. In recent years, with the development of various omics and synthetic biology technologies, the biosynthetic pathways of sesquiterpene lactone compounds of different structural types have gradually been resolved. Among them, the researches on germacrene-derived sesquiterpene lactones are relatively more than others. Therefore, this article focused on the germacrene-derived sesquiterpene lactone biosynthesis pathways and their key enzyme genes, which can lay the foundation for in-depth analysis of sesquiterpene lactone biosynthetic pathways, functional gene mining and heterologous synthesis of active ingredients.

Bioactive metabolites in of Ginkgo biloba leaves: variations by seasonal, meteorological and soil / Metabólitos bioativos em folhas de Ginkgo biloba: variações sazonais, meteorológicas e do solo

Abstract Ginkgo biloba is a traditional Chinese herbal medicine containing multiple components that contribute to its notable bioactivities. Variations of seasonal, meteorological and planting soil on the phytochemicals contents in G. biloba leaves due to the effects of growth meteorological and soil parameters were investigated in this study. The leaves of G. biloba were collected from different months and place in Zhejiang province, the contents of flavones (quercetin, kaempferol and isorhamnetin) and terpene lactones (bilobalide, ginkgolides A, B and C) were quantified by high performance liquid chromatography (HPLC) and the evaporative light scattering detector (ELSD) method. The established methods were validated with good linearity, precision, repeatability, stability, and recovery. Comprehensive analysis suggested the proper harvest time for G. biloba was in October of Zhejiang province. The result of correlation analysis with meteorological factors shows that the temperature and precipitation have non-significant effect on the main components of G. biloba. In addition, the type and content (Mn and Zn) of the soil showed significantly effect on the content of flavonoids and terpene lactones. This study enriched the knowledge on the development and utilization value of the G. biloba leaves and was useful for determining the optimal harvest time and growing condition.

Resumo Ginkgo biloba é um fitoterápico tradicional da medicina chinesa que contém vários componentes que contribuem para suas notáveis bioatividades. Variações sazonais, meteorológicos e de plantio do solo sobre os teores fitoquímicos em folhas de G. biloba, devido aos efeitos do crescimento meteorológico e parâmetros do solo, foram investigadas neste estudo. As folhas de G. biloba foram coletadas em diferentes meses e na província de Zhejiang, os teores de flavonas (quercetina, kaempferol e isorhamnetina) e lactonas terpênicas (bilobalida, ginkgolídeos A, B e C) foram quantificados por Cromatografia Líquida de Alta Eficiência (CLAE) e pelo método do detector de dispersão da luz por evaporação. Os métodos estabelecidos foram validados com boa linearidade, precisão, repetibilidade, estabilidade e recuperação. Uma análise abrangente sugeriu que o tempo de colheita adequado para G. biloba foi em outubro na província de Zhejiang. O resultado da análise de correlação com fatores meteorológicos mostra que a temperatura e a precipitação não têm efeito significativo sobre os principais componentes de G. biloba. Além disso, o tipo e composição (Mn e Zn) do solo apresentaram efeito significativo sobre o teor de flavonoides e lactonas terpênicas. Este estudo enriqueceu o conhecimento sobre o valor de desenvolvimento e utilização das folhas de G. biloba e foi útil para determinar o melhor tempo de colheita e condição de crescimento.

Computational prediction of active sites and ligands in different AHL quorum quenching lactonases and acylases

Rice tungro is a serious viral disease of rice resulting from infection by two viruses, Rice tungro bacilliformvirus and Rice tungro spherical virus. To gain molecular insights into the global gene expression changes inrice during tungro, a comparative whole genome transcriptome study was performed on healthy and tungroaffected rice plants using Illumina Hiseq 2500. About 10 GB of sequenced data comprising about 50 millionpaired end reads per sample were then aligned on to the rice genome. Gene expression analysis revealedaround 959 transcripts, related to various cellular pathways concerning stress response and hormonal homeostasis to be differentially expressed. The data was validated through qRT-PCR. Gene ontology and pathwayanalyses revealed enrichment of transcripts and processes similar to the differentially expressed genes categories. In short, the present study is a comprehensive coverage of the differential gene expression landscapeand provides molecular insights into the infection dynamics of the rice-tungro virus system

Effect of ginkgo diterpene lactones on hypoxia-induced apoptosis and angiogenesis of human umbilical vein endothelial cells and its mechanisms / 第二军医大学学报

Objective To investigate the effect of ginkgo diterpene lactones on hypoxia-induced apoptosis and angiogenesis of human umbilical vein endothelial cells (HUVECs) and the related molecular mechanisms. Methods HUVECs were cultured under hypoxia for 24 h, and then treated with ginkgo diterpene lactones (low-dose: 6.25 mg/L and high-dose: 25.00 mg/L). MTT assay was used to detect the cell activity. Flow cytometry was used to detect the apoptosis of HUVECs. Transwell assay was employed to detect the migration of HUVECs. Quantitative real-time polymerase chain reaction and Western blotting were used to test the expression levels of mRNA and protein of hypoxia-inducible factor 1α (HIF-1α), apoptosis-related genes (B-cell lymphoma 2［Bcl-2］and B-cell lymphoma 2-related X protein［Bax］), and angiogenesis-related genes (vascular endothelial growth factor［VEGF］and transforming growth factor β［TGF-β］). Results Compared with the normal group, the HUVEC activity was significantly decreased after exposed to hypoxia for 24 h (P＜0.05), and the apoptosis rate of HUVECs and mRNA and protein expression levels of HIF-1α were significantly higher (all P＜0.05). The cell activity and migration ability of HUVECs were significantly higher in the low- and high-dose ginkgo diterpene lactones groups than those in the hypoxia group (all P＜0.05), and the apoptosis rates of HUVECs were significantly lower than those in the hypoxia group (both P＜0.05). Meanwhile, the cell activity and migration ability were significantly higher in the high-dose group than those in the low-dose group (both P＜0.05), and the apoptosis rate was significantly lower than that in the low-dose group (P＜0.05). The mRNA and protein expression levels of HIF-1α and Bax were lower in the low- and high-dose ginkgo diterpene lactones groups than those in the hypoxia group, and the mRNA and protein expression levels of Bcl-2, VEGF and TGF-β were higher than those in the hypoxia group; and the expression changes of the above genes were more significant in the high-dose group. Conclusion Ginkgo diterpene lactones can improve hypoxia-induced apoptosis and angiogenesis of HUVECs by regulating the expression levels of HIF-1α and apoptosis- and angiogenesis-related genes, and it might be used as a new agent to treat anoxic vascular diseases.

The crucial role of metabolic regulation in differential hepatotoxicity induced by furanoids in Dioscorea bulbifera / 中国天然药物

Diterpenoid lactones (DLs), a group of furan-containing compounds found in Dioscorea bulbifera L. (DB), have been reported to be associated with hepatotoxicity. Different hepatotoxicities of these DLs have been observed in vitro, but reasonable explanations for the differential hepatotoxicity have not been provided. Herein, the present study aimed to confirm the potential factors that contribute to varied hepatotoxicity of four representative DLs (diosbulbins A, B, C, F). In vitro toxic effects were evaluated in various cell models and the interactions between DLs and CYP3A4 at the atomic level were simulated by molecular docking. Results showed that DLs exhibited varied cytotoxicities, and that CYP3A4 played a modulatory role in this process. Moreover, structural variation may cause different affinities between DLs and CYP3A4, which was positively correlated with the observation of cytotoxicity. In addition, analysis of the glutathione (GSH) conjugates indicated that reactive intermediates were formed by metabolic oxidation that occurred on the furan moiety of DLs, whereas, GSH consumption analysis reflected the consistency between the reactive metabolites and the hepatotoxicity. Collectively, our findings illustrated that the metabolic regulation played a crucial role in generating the varied hepatotoxicity of DLs.

Three new polyoxygenated bergamotanes from the endophytic fungus Penicillium purpurogenum IMM 003 and their inhibitory activity against pancreatic lipase / 中国天然药物

Purpurolides D-F (1-3), three new polyoxygenated bergamotanes bearing a 6/4/5/5 tetracyclic ring system, were isolated from the endophytic fungus Penicillium purpurogenum IMM 003. Their structures were unambiguously elucidated based on extensive spectroscopic data analyses, C NMR chemical shifts calculations coupled with the DP4+ probability method, and the calculated and experimental electronic circular dichroism (ECD) spectra. Compounds 1-3 showed significant inhibitory activity against pancreatic lipase (PL). The result highlights that the presence of 3-hydroxylated decanoic acid moiety at C-14 is important for increasing the inhibition potency against PL.

In vivo anti-hyperuricemic activity of sesquiterpene lactones from Lychnophora species

ABSTRACT Hyperuricemia is the main cause of gout, an inflammation induced by uric acid deposition in joints. Drugs available present side effects, so there is a need for new treatment alternatives. Lychnophora species are used in folk medicine to treat inflammation, rheumatism and muscle pain. Goyazensolide (10 mg/kg), eremantholide C (25 mg/kg) and lychnopholide (25 mg/kg), sesquiterpene lactones isolated from Lychnophora species were previously studied and showed anti-hyperuricemic effects in mice. However, the mechanisms of this effect were not elucidated. The methodology of this study consisted in treatment of hyperuricemic-induced rats, and comparison between control groups, clinically used anti-hyperuricemic drugs and sesquiterpene lactones. Urine and blood were collected for uric acid quantification. Xanthine oxidase inhibition was measured in liver homogenates. Results showed that all evaluated sesquiterpene lactones presented anti-hyperuricemic activity at the doses of 5 and 10 mg/kg and can act through one or both mechanisms, depending on the dose administrated. Goyazensolide and lychnopholide at dose of 5 mg/kg showed important uricosuric effect. Goyazensolide and lychnopholide at dose of 10 mg/kg, and eremantholide C (5 and 10 mg/kg) presented notable inhibition of hepatic xanthine oxidase activity and uricosuric effect. Thus, these sesquiterpene lactones are promising hypouricemic agent to treat hyperuricemia and gout.

Chemical constituents of Pestalotiopsis microspora HF 12440

Phytochemical investigation on the liquid media of Pestalotiopsis microspora HF 12440, an endophytic fungusisolated from the stem of Artocarpus heterophyllus, led to the isolation of three lactones, (+)-acetylpestalotin (1),(−)-pestalotin (2), (6S,7S,8R)-hydroxypestalotin (3) and a lignan, (+)-pinoresinol (4). Interestingly, one of them (1)is a new compound, while compound 4 is reported for the first time from endophytic fungi. Structures of the isolatedcompounds were determined based on spectroscopic data, including MS, NMR 1D and 2D. Cytotoxic values of thecrude extract and all isolated compounds were evaluated against P388 cells, showing that compound 3 was the mostactive with IC50 3.34 μg/ml. Overall, this is the first phytochemical report of endophytic fungi isolated from the hostplant Artocarpus

Relationship Between Parameters of Development and Functional Compounds of Yacon Leaves

Abstract Yacon is a tuberous root from the Andean region being increasingly grown across the world due to the low caloric values of their roots and the functional properties of extractions of its leaves. However, there is still a gap in the knowledge of how the plant develops, and if physical and chemical changes are noticed regarding maturity. The subject of this work was to investigate the dynamics of a group of parameters and to establish correlations across them. To achieve this goal, leaves collected from three different parts of the plant in three months were assessed regarding size (plant height and leaf area), color and the presence of sesquiterpene lactones. Different methods of extraction were also studied and the total phenolic content was analyzed. The analysis of the results revealed different patterns in the quantity, size and leaf distribution according to the age of the plant. From three different methods of extraction studied, decoction seems to be the most appropriate one to obtain phenolic compounds with greater yield and safety. Besides, the greenness of yacon leaves showed a linear correlation to the total phenolic content, being a simple and promising estimator of the corresponding bioactive group. Sesquiterpene lactones were identified in all extracts, indicating the ease of their extractability even in simple procedures of tea-making.

Simultaneous determination of four sesquiterpene lactones in the whole herb of Elephantopus scaber Linn / 国际药学研究杂志

Objective: To establish an ultra-performance liquid chromatography(UPLC)method for the simul-taneous determination of four sesquiterpene lactones, deoxyelephantopin, isodeoxyelephantopin, isoscabertopin and sca- bertopin in Elephantopus scaber Linn.. Methods: The UPLC conditions were as follows: the separation was performed using an Acquity UPLC BEH C18(2.1 mm × 100 mm, 1.7 μm)column, the temperature of the column was maintained at 40℃, and the detection wavelength was 220 nm. The mobile phase was the water and acetonitrile solution in a binary gra- dient elution at the flow rate of 0.3 ml/min. Results: The four components, deoxyelephantopin, isodeoxyelephantopin, isoscabertopin and scabertopin were separated well under the UPLC conditions and showed a good linearity within the ranges of 0.383-76.500(R2=0.9999), 0.775-155.0(R2=0.9999), 0.507-101.5(R2=0.9999), and 0.850-(R2=0.9997), respectively. Their average recovery(n=6)was 102.31%, 103.87%, 102.66% and 105.68%, RSD was 1.20%, 2.00%, 1.24% and 1.73%, respectively. The four components were validated by the simultaneous termination in eleven batches of E. scaber samples from different areas in the present study. Conclusion: lished UPLC method could be used for the determination of four sesquiterpene lactones, deoxyelephantopin, elephantopin, isoscabertopin and scabertopin in E. scaber. The present results provide a reference for further velopment and quality evaluation of E. scaber.

Establishment of QAMS method with dehydroandrographolide as internal reference substance for quality control of Andrographis Herba / 中国中药杂志

To solve the problems of the poor resolution of chromatographic separation,the weak durability of the relative correction factors,and the low accuracy of content determination results in the quantitative analysis of multi-components by single-marker( QAMS) method with andrographolide as the internal reference substance in the existing research of Andrographis Herba,a new QAMS method using dehydroandrographolide as the internal reference substance was established for the first time in this study. This new method can be used to simultaneously determine four diterpene lactones,including andrographolide( A),neoandrographolide( B),14-deoxyandrographolide( C),and dehydroandrographolide( S) through the optimization of chromatographic conditions and systematic investigation of methodology. At the present HPLC chromatographic conditions,four components could be well separated( R > 1. 5),and the methodology validations could satisfy the requirement of quantitative analysis. The relative correction factors( RCFs) of fA/S,fB/S,fC/S were determined as 0. 65,0. 54,0. 78,respectively. The relative standard deviations( RSDs) of their RCFs ranged between 1. 3%-5. 1%,0. 25%-0. 33%,0. 070%-0. 15%,0. 070%-0. 22%,respectively with three brands of HPLC instruments,five brands of C18 column,different flow rates( 0. 9,1. 0,1. 1 m L·min~(-1)),and different column temperatures( 25,30,35 ℃),indicating good durability of the RCFs. The relative retention value( RRV) method was used to locate the chromatographic peak of the components to be determined.The RRVs of rA/S,rB/S,and rC/Swere 0. 44,0. 86,0. 97,respectively. The RSDs of the RRVs ranged between 0. 030%-1. 6% with different HPLC instruments and columns,showing accurate peak location. The present QAMS method and the external standard method( ESM)were both used to determine the contents of four diterpene lactones from Andrographis Herba( 6 batches of medicinal materials and 18 batches of cut crude drugs). The relative errors of the determined content results between two methods were less than 2. 0%. It demonstrated that there was no significant difference in content results between these two methods,indicating good accuracy of the present QAMS method. Therefore,in this study,an accurate and highly durable QAMS method using dehydroandrographolide as the internal reference substance was established for simultaneous determination of four diterpene lactones. This method could be used to effectively control the quality of Andrographis Herba and provide technical basis for the formulation of traditional Chinese medicine industry standard and improvement of the Chinese Pharmacopoeia standard of Andrographis Herba.

Quality Evaluation of Andrographis Herba Based On Electronic-eye Technique / 中国实验方剂学杂志

Objective: Taking electronic-eye (visual analyzer) technique,based on the powder color of Andrographis Herba,to investigate the applicability of electronic-eye technique and evaluate the quality of Andrographis Herba with different commercial specifications. Method: HPLC was employed to determine contents of andrographolide,dehydroandrographolide,14-deoxyandrographolide,neoandrographolide in 50 batches of Andrographis Herba with different commercial specifications(stems,leaves and aerial parts).Color of these samples were measured by electronic-eye technique.The data were analyzed by principal component analysis(PCA) and Pearson correlation analysis.The ability of electronic-eye to distinguish the different commercial specifications of Andrographis Herba was investigated and the correlation of chroma space system parameters (L*,a*,b*) with active components was investigated. Result: There was remarkable difference in contents of 4 diterpenoids in Andrographis Herba from different parts,their contents in leaves was the highest,followed by the aerial parts(mixture of stems and leaves),and their contents in stems was the lowest.The results of PCA was divided into two classes,namely the stem part,leaf and aerial parts,indicating that electronic-eye could be used to distinguish the quality of Andrographis Herba.The correlation results showed that there were significant negative correlation(PL*(lightness value) and the contents of andrographolide,dehydroandrographolide,14-deoxyandrographolide,neoandrographolide and the total content of these 4 components.In addition,L* of samples that did not conform to the lower limit of determination in the 2015 edition of Chinese Pharmacopeia was ≥ 69.5,and the L* of more than 90% of the samples in accordance with the requirements was Conclusion: Electronic-eye technique provides a new method and idea for the quality evaluation of Andrographis Herba.

Anti-inflammatory Activity of Standardized Fraction from Inula helenium L. via Suppression of NF-κB Pathway in RAW 264.7 Cells

Inula helenium L. is rich source of eudesmane-type sesquiterpene lactones, mainly alantolactone and isoalantolactone, which have the various pharmacological functions. In this study, we examined the inhibitory effects of nitric oxide (NO) production of hexane, methylene chloride, ethyl acetate, butanol, and water fractions from I. helenium and investigated the anti-inflammatory effect of hexane fraction of I. helenium (HFIH) in LPS-induced RAW 264.7 cells. Quantification of alantolactone and isoalantolactone from HFIH was carried out for the standardization by multiple reaction monitoring using triple quadrupole mass spectrometer. HFIH significantly inhibited inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2) protein as well as their downstream products NO and prostaglandin E₂ (PGE₂) in LPS-stimulated RAW 264.7 cells. Moreover, HFIH suppressed NF-κB transcriptional activity by decreasing the translocation of p65 to the nucleus. The in vivo study further confirmed that HFIH attenuated the paw edema induced by carrageenan in an acute inflammation model. These findings suggest that HFIH may be useful as a promising phytomedicine for inflammatory-associated diseases.

Main chemical constituents of Shengui Capsules based on UHPLC-Q-Orbitrap HRMS / 中草药

Objective: In order to study the chemical profile of Shengui Capsules, a novel ultra high performance liquid chromatography-quadrupole/orbitrap high resolution mass spectrometry (UHPLC-Q-Orbitrap HRMS) was applied to establish a method to recognize and classify the ingredients accurately and rapidly. Methods: ACQUITY UPLC® BEH C18 chromatographic column (100 mm × 2.1 mm, 1.7 μm) was employed and the separation was performed with the mobile phase consisting of acetonitrile-0.1% formic acid aqueous solution. The information of accurate mass and multistage fragment ions were obtained by the novel “monitored simultaneously for positive and negative ions, Full MS scan and automatic trigger secondary mass spectrometry” mode of Q-Orbitrap MS technology. Then, the complex chemical constituents would be identified by comparing the relative retention time and the mass data of the reference substance, and consulting the reference literature or Mass Bank, Chemical Book network database at the same time. Results: Forty chemical components were finally identified in this study, including the 16 tritepene saponins, 6 organic acids, 13 lactones, and 5 other classes. Conclusion: The method established in this study could be used to rapidly, accurately, and systematically identify the main chemical constituents of Shengui Capsules. Moreover, this study will lay a scientific preliminary foundation for the further pharmacodynamic material basis, quality control improvement and the rational clinical use.

Induction of p450 enzymes by derivative of parthenolide / 中草药

Objective: In this study, primary cultured human hepatocytes were used to study the induction of P450 enzyme by the sesquiterpene lactone derivative ACT001, which provided reference for the clinical use of ACT001. Methods Three batches of frozen primary human hepatocytes were inoculated and cultured, and CYP1A2, CYP2B6 and CYP3A4 were induced by ACT001. Real-time fluorescence quantitative PCR was used to determine the mRNA expression level of P450 enzyme, and the activity of P450 enzyme was determined by LC-MS/MS method. Results The expression level of P450 enzyme mRNA and the activity of P450 enzyme showed that the P450 enzyme induction model was successfully established. Compared with the control group, the CYP1A2 mRNA expression level and enzyme activity of ACT001 1 μmol/L and 6 μmol/L group showed no significant changes. The mRNA expression level of CYP1A2 in ACT001 30 μmol/L group was significantly decreased, and the enzyme activity was decreased, but not as significantly as that of mRNA. With the increase of ACT001 concentration, the expression level of CYP2B6 mRNA was gradually increased. Compared with the control group, the expression level of CYP2B6 mRNA in the ACT001 group at 30 μmol/L was significantly increased, which was seven times higher than that in the control group, and the increase of enzyme activity was four times higher than that in the control group, which was 40% higher than that in the phenobarbital sodium induction multiple. Compared with the control group, the CYP3A4 mRNA expression level of cells in the ACT001 1 μmol/L group was significantly increased, which was four times higher than that of the control group, but did not reach 40% of the positive inducer rifampicin, and the CYP3A4 mRNA expression level was decreased gradually with the increase of ACT001 concentration. At the same time, there was no significant increase in CYP3A4 enzyme activity after ACT001 administration at different concentrations, which was less than two times of that in the control group. Conclusion The data indicated that ACT001 had no induction potential for CYP1A2 and CYP3A4, and had potential for CYP2B6 induction. In combination with CYP2B6 substrates, it should be avoided in clinical combination therapy to reduce adverse reactions caused by P450-mediated drug drug interaction.

Effect of γ-lactones and γ-lactams compounds on Streptococcus mutans biofilms

Abstract Considering oral diseases, antibiofilm compounds can decrease the accumulation of pathogenic species such as Streptococcus mutans at micro-areas of teeth, dental restorations or implant-supported prostheses. Objective To assess the effect of thirteen different novel lactam-based compounds on the inhibition of S. mutans biofilm formation. Material and methods We synthesized compounds based on γ-lactones analogues from rubrolides by a mucochloric acid process and converted them into their corresponding γ-hydroxy-γ-lactams by a reaction with isobutylamine and propylamine. Compounds concentrations ranging from 0.17 up to 87.5 μg mL-1 were tested against S. mutans. We diluted the exponential cultures in TSB and incubated them (37°C) in the presence of different γ-lactones or γ-lactams dilutions. Afterwards, we measured the planktonic growth by optical density at 630 nm and therefore assessed the biofilm density by the crystal violet staining method. Results Twelve compounds were active against biofilm formation, showing no effect on bacterial viability. Only one compound was inactive against both planktonic and biofilm growth. The highest biofilm inhibition (inhibition rate above 60%) was obtained for two compounds while three other compounds revealed an inhibition rate above 40%. Conclusions Twelve of the thirteen compounds revealed effective inhibition of S. mutans biofilm formation, with eight of them showing a specific antibiofilm effect.

Simultaneous Determination of Four Lactones Ingredients Content in Yinxingye Tablets by Quantitative Analysis of Multi-Components by Single-Marker / 中国药学杂志

OBJECTIVE To determine simultaneously four lactones ingredients content in Yinxingye tablets by quantitative analysis of multi-components by single-marker (QAMS), and to verify the application value of this method in the quality control of Yinxingye tablets. METHODS With Ginkgo esters as the indexes,the relative correction factor(RCFs) between bilobalide and ginkgolides A,B,C were established. The content of bilobalide in Yinxingye tablets were determined by external standard method,the content of ginkgolides A,B,C in Yinxingye tablets was calculated by the relative correction factor. The external standard method and QAMS determined contents of 4 lactones, Rationality, feasibility and repeatability of the QAMS method were verified by comparing the results obtained from two different methods. RESULTS There was no significant differences between the calculated value and estimated value on QAMS and external standard method of the 5 batches of Yinxingye tablets,the results showed that the RCF was authentic. CONCLUSION The established correction factor is good repeatability,and could be used for quantitative analysis and quality evaluation of multiple components in Yinxingye tablets.

Rapid screening and identification of sesquiterpene lactones in Kudiezi injection based on high-performance liquid chromatography coupled with linear ion trap-orbitrap mass spectrometry / 中国天然药物

Sesquiterpene lactones are considered as the major active compounds in Kudiezi injection in virtue of their special structures and activities. Herein, an analytical method was developed for rapid screening and identification of sesquiterpene lactones in Kudiezi injection using high-performance liquid chromatography coupled with linear ion trap-orbitrap mass spectrometry (HPLC-LTQ-Orbitrap) in negative ion mode. First, two sesquiterpene lactone reference standards were analyzed to obtain their characteristic ESI-MS/MS fragmentation patterns. Second, based on extracted ion chromatography (EIC) data-mining method and characteristic fragmentation pathways analysis, sesquiterpene lactones in Kudiezi injection were rapidly screened and identified. Finally, an important parameter Clog P was adopted to discriminate the isomers of sesquiterpene lactones. As a result, 50 sesquiterpene lactones were characterized, including 9 sesquiterpene lactone aglycones, 39 sesquiterpene lactone glycosides, and 2 amino acid-sesquiterpene lactone conjugates. Among them, 13 compounds were tentatively identified as new compounds. The results demonstrated that the established method would be a rapid, effective analytical tool for screening and identification of sesquiterpene lactones in the complex system of natural medicines.